Poultry Disease Manual

Poultry Disease Manual
Pro/. Antonio Zanella
Contents
4
1 PREFACE
IM M UN ITY IN BIRDS: GENERAL CONCEPTS
NEWCASTLE DISEASE
AVIAN PNEUMOVIRUSES
AVIAN INFLUENZA
INFECTIOUS BRONCHITIS
INFECTIOUS LARYNGOTRACHEITIS
26
FOWL POX
13
INFECTIOUS BURSAL DISEASE
Neoplastic Diseases
- MAREK'S DISEASE
- LEUKOSIS/SARCOMA GROUP
- RETICULOENDOTHELIOSIS
CHICKEN INFECTIOUS ANAEMIA
4-2 REOVIRUS INFECTIONS
44
rn ADENOVIRUS INFECTIONS
M AVIAN ENCEPHALOMYELITIS
PARATYPHOID INFECTIONS
54
m PULLORUM DISEASE AN D FOWL TYPHOID

COLIBACILLOSIS
INFECTIOUS CORYZA
FOWL CHOLERA
62
NECROTIC ENTERITIS
m RiEMERELLA ANATIPESTIFER
Mycoplasmosis
65
69
70
- MYCOPLASMA GALL1SEPTICUM INFECTION

- MYCOPLASMA SYNOVIAE INFECTION
- MYCOPLASMA MELEAGRIDIS INFECTION
COCCI DIOSIS
. VACCINE ADMINISTRATION SYSTEM
79
81
5
FACTORS INFLUENCING VACCINATION

M DIAGNOSIS OF DISEASE
si- DIAGNOSIS OF SOME MOST IMPORTANT POULTRY DISEASE
Preface
I am h o n o u re d th at F A T R O re q u e ste d m e to w rite th is (sh o rt)
I
m anual o n in fectious diseases, m ostly o f intensively-reared dom estic

poultry.
T h is review is n o t com pletely o rig in al, b u t is ob v io u sly a m ixture
o f d a ta re p o rte d by m an y e m in e n t C o lle a g u e s, R e se a rc h e rs o r
P ractition ers, w ith th e ad d itio n o f my ow n p e rso n al ob servation s,
d erived from m ore th an 5 0 years o f intensive experien ce in poultry
pathology. M y on ly m erit, I rep eat, h as been to collect an d u p d ate
the know ledge o n the variou s diseases regarding their epidem iology,
c lin ica l sig n s, le sio n s, d ia g n o stic s a n d , o b v io u sly , th e ir c o n tro l
th ro u gh biosecu rity m easu res, treatm en ts an d vaccin ation s.
1 dedicate this b o o k to m y beloved wife, children an d grandchildren,
w ho bore with m e throughout this long period o f my professional life.
I w ish to p a r tic u la r ly th a n k m y C o lle a g u e s D r. F. M c lio ta ,
Dr. C . T assi, Dr. A . V olorio, M r. P. M ilani and M rs. D ebora Bcrtozzi
fo r th eir valu ab le help.
Immunity in birds: general concepts

Harderian gland
T h e im m u n e sy ste m o f th e b ir d s I
c o n sis ts o f tw o p rim a ry ly m p h o id
Payer's patches
organ s, the thym us an d the b u rsa o f
bursa
of Fabricius
F a b r ic iu s (B F ), b u t a ls o o f m an y
seco n d ary
ly m p h o id
o rgan s
or
structures, which include bone-marrow,

thymus
spleen, H arderian gland, Peyers patches,

M eckel diverticulum , caecal tonsils and
oth er lym ph oid aggregates distribu ted
cecal tonsils
in the body (birds lack the equivalen t

spleen
o f m am m alian lymph nodes).

In the primary organs, w hich d evelop
d u rin g the em bryo stage, a d ifferen t
Meckel diverticulum
tiatio n of the germ in al im m u n e cells

bone marrow
in to two types o f ly m p h ocy tes takes

place, T-cells in the thymus an d B-cells
in the BF. T hese cells produce a specific
im m u n e r e s p o n s e a g a in st a n tig e n s
which are extran eous to the organism .
In th e secondary stru ctu res, b e sid e s
lymphocyte storage areas, there are the differentiation sites for other
Diagram of the Immune System of birds
im m u n e cells, such as m onocytes and m acrop h ages, granulocytes, |

j k ('j]er (JC)
natu ral killer (N K ) cells, which are p ro d u ce d in the J
bone-m arrow an d generally play a non-specific role in im m unity.
B'lyrnphocytes, after tran sform ation into plasm ocytes, are responsible
| for im m unoglobu lin or antibody production, in response to antigen
stim u lation .
T'bmpfioQtes are re sp o n sib le for cell-m ediated im m u n ity and for
regulation o f the im m u ne system reactions, in cluding the activation
o f B-cells.
Macrophages (derived from c irc u latin g m o n o cy tes), granulocytes,
N K and K cells are involved non-specifically in im m u n e reaction s
by phagocytosis or by cytotoxicity. M acrop h ages are also im p o rtan t
in an tigen p rocessin g an d p resen tin g cell to T and B lym phocytes.
Immunoglobulins (Ig) or an tib o d ies are glycoprotein m olecules, able
to react specifically, both in vivo and in
originally in du ced their p rod u ctio n .
with the antigens which
In bird s, three m ain classes o f Ig have b een rep orted :

IgM , w hich ap p e a r very so o n , 2-4 days afte r th e in itial im m u n e
stim u latio n , reach ing their peak co n cen tratio n a t th e 9 '1' day; they
represent the first im m u n e b arrier to in fection ; IgG (or, better, IgY,
sh ow in g so m e structural d ifferen ces to m am m alian IgG), w hich are
th e m ain Ig in th e se ru m (7 0 -8 0 % ) a n d th e m ain e ffe c to rs o f
h u m o ral im m unity; they are th e on ly an tib o d ies ab le to tran sfer to
th e yolk-sac an d provid e system ic im m u n ity to the n ew born chick;
/gAS, which are the Ig present in m ucosal secretions and in biological
Small lymphocyte
flu id s, as bile an d resp on sible fo r m u co sal im m unity; the existence

o f IgD an d IgE, as in m am m als, is very likely, b u t little is know n on
th is m atter.
H U M O R A L IM M U N IT Y
H u m o ra l im m u n ity is su b d iv id e d in to active a n d passive.
A ctive im m u n ity is triggered by con tact with the antigen, pathogen
o r v a c c in e a n d is c h a ra c te riz e d , a fte r a v a r ia b le la te n t p e rio d
(3 to 10 days), by an in itial rap id s h o r t in crease in IgM , follow ed
by a slow er increase in IgG and IgA, w hich p ersists lon ger (prim ary
respon se). S u b se q u e n t c o n tact w ith th e sa m e p ath o gen o r antigen
in duces a higher, m ore rapid and long-lasting p rod u ction o f specific
IgG a n d IgA (seco n d ary resp on se). T h e rap id ity o f the se c o n d ary
response is d u e to the presence o f m em ory lym phocytes", produced
d u rin g the p rim ary re sp o n se ; fo r th is reason it is also n am ed the
an am n estic respon se. R ep eated vaccin ation s (hyperim m unization
Plosmo-cell
o r booster effect) enable the attainm ent o f high specific antibody levels.
P assive im m u n ity is provided alm o st exclusively by IgG , transferred
fro m th e b lo o d stream to th e ovary a n d th en tran sm itte d to the
chicks via the yolk-sac; the level o f an tibodies in the yolk sac generally
correlates w ith th e m o th ers an tib o d y level.
D u rin g passage o f the egg through the oviduct, the album en acquires
rath er low levels o f IgM an d IgA, w hich p ass th rou gh the am n iotic
flu id to th e g u t, p ro v id in g te m p o rary p ro te ctio n to th e m u cosa.
T h e level o f m atern al an tib o d ies in the progeny norm ally decreases
in a b o u t 15-20 d ays; th e rate o f d e cre ase is p artly lin k ed to the
d egree o f grow th o f th e chick.
T h e p ro tec tio n p rovid ed by passive a n tib o d ie s v aries, d e p e n d in g
o n the d isease: fo r exam ple, it is stro n g again st in fectiou s an aem ia,
encephalom yelitis, in fectiou s b u rsa l d isease, fairly effective again st
N e w ca stle d is e a s e a n d p o o r a g a in s t m u c o sa l d is e a s e s, su c h as
in fe c tio u s b ro n c h itis a n d lary n g o trac h e itis a n d ag a in st M a re k s
Large lymphocyte
d isease.
CELL-MEDIATED !MMl)NITY
C ell-m ed ia ted im m u n ity (C M I) is c o n tro lle d by T -lym phocytes,
which d o n o t produ ce antibodies, but operate destroying the target
directly o r indirectly, in du cin g o th e r accessory cells to destroy it by
in term ed iation o f so lu b le factors, the in terleukines. T-lymphocytes
are d iv id ed in to three su b p o p u la tio n s: T-helper, T -su p pressor and
T-cytotoxic cells, b e sid e s m em ory cclls:
T-helper cells h av e re g u la to ry activity, m o d u la tin g th e im m u n e
resp on se an d intervening in the coop eration betw een cell-m ediated
Monocyte
a n d h u m o ral im m unity;
T-suppressor cells have re g u lato ry activity, lim itin g th e a n tib o d y
resp on se, w hen th is te n d s to becom e excessive;
T-cytotoxic c c lls h av e e ffe c to r activity , a tta c k in g a n d d e stro y in g
in fected o r alte re d target cells. C M 1 is o f p rim ary im p o rta n c e in
im m u n e d e fe n c e s a g a in s t m ic r o o r g a n is m s w ith in tra c e llu la r
p arasitism (viru ses, so m e b a c te ria , p ro to zo a n s, etc.). In gen eral,
w hile h u m oral im m u nity can be m easured q u ite easily by checking
in vitro th e level o f an tib o d ie s in the se ru m , it is m ore d iffic u lt o r
complex, in a routine check, to evaluate d ie degree of mucosal and CM1.
M oreover, th e d egree o f p ro te ctio n p rovided by a vaccin e can n o t
be always strictly correlated to the level o f an tib o d ie s in th e serum .
VACCINES
L iv e a tte n u a te d v a c c in e s are p rep ared w ith:
1) naturally apath ogenic m icroorganism s (turkey herpesvirus, Newcastle
Basophil Granulocyte
disease virus strain s N D V 6 / 1 0 o r H itch n er B l) ;

2) m icroorganism s, w hose pathogenicity- has been artificially reduced
o r rem oved by serial p assag es th rou gh an ap p ro p ria te su b stratu m ,
such as em bryon ated egg o r tissue-cultures (infectious bronchitis virus,
infectious bursal disease virus an d m any o th e r vaccin al viruses);
3) other techniques, fo r the m o m en t m ainly experim ental (deletion,
exp ressio n o f an tigen in live ap ath o g e n ic vectors).
T h e artificial atten u ation o f m icroo rgan ism s is som etim es u n stable
an d , after so m e back-passages in the natural host, it m ay b e reduced
(laiyngotracheitis, infectious bursal disease, infectious bronchitis viruses, etc.).
A tte n u a te d v a cc in es a re n o rm ally a d m in iste re d to b ird s by the
natural route, like the p ath ogen ic m icroorganism s, b u t they induce
very low o r n o sy m p to m s o f d isease.
T h e c o m m o n rou te o f a d m in istra tio n in th e field is by d rin k in g
w ater o r spray or, individually, by eye o r n asal-drop ad m in istratio n
o r wing-w eb in je ctio n . P ro tectio n d u e to the v a cc in atio n is o fte n
Heterophil Granulocyte
provided q u ite quickly after adm inistration (local im m unity system),
alth o u g h a n tib o d ie s a p p e a r in th e se ru m on ly a fte r 10-15 days.

F or so m e diseases, ju st o n e vaccin ation a t a variable age, d epe n d in g
o n th e v a c c in e , is s u f fic ie n t to p ro te c t b ir d s fo r a lo n g rim e
(M areks disease, fowl pox, laryngotracheitis, encephalomyelitis, infectious
anaemia) o r for the period w hen they are susceptible (infectious bursal
disease).
F o r o th e r d ise a s e s, re v a c c in a tio n s o r b o o ste r s a re re q u ire d to
strengthen the protection (Newcastle disease, infectious bronchitis, etc.).
F or so m e p ath o gen s, the b ird s are su scep tible on ly u p to 2-3 weeks
Eosinophil granulocyte
o f age a n d it is su ffic ie n t to vaccin ate p a re n t m e n s befo re g o in g

in to lay (infectious anaem ia, encephalomyelitis) to p rotect th e y ou n g
progeny. Live v a cc in e s m ay p re se n t so m e risk o f c o n ta m in a tio n
w ith fo re ig n a g e n ts p re se n t in th e su b stra te (leukosis, infectious
anaem ia, etc.), o r accid en tally in tro d u c e d d u rin g th e ir p ro cessin g
(or m anufacturing). In any event, the use o f eggs from well-controlled
S P I- flo c k s an d th e a d o p tio n o f severe p u rity c o n tro ls, firstly o f
m a ste r se e d s, b u t a ls o o f e a c h b a tc h o f v a c c in e , a rc e sse n tia l.
S o m e live vaccin es are also p rep ared w ith a tte n u a te d b acteria o r
p ro to z o a n s (fo r in sta n c e , safm onedas, mycoplasmas a n d coccidia).
In activated vaccin es are prepared w ith m icroo rgan ism s, o r p art o f
th e m , w h ich h ave lo st th e ir v iru le n c e a n d ab ility to re p lic ate
an d sp read th rou gh th e organ ism . H ow ever, their tm m unogenicity
re m a in s in ta c t. S u c h v a c c in e s a re a d m in iste r e d by in d iv id u a l
in jection , in tram u scu larly o r su bcu tan eou sly .
Macrophage
T h e y are o fte n a sso c ia te d w ith a d ju v a n ts, w h ich stre n g th e n the

im m u n e re a c tio n . T h e a d ju v a n ts m ain ly u se d are m in eral oil
(em ulsion) o r alu m iniu m hydroxide. T h e stren gth en in g o f an tibo dy
p ro d u c tio n a n d , partially, o f cell-m ed iated reactivity, is d u e to a
g rad u al release o f an tig en (d e p o t effect) an d local irritatio n , with
a sp e c ific ac tiv a tio n o f ly m p h ocy tes. T h e im m u n ity p ro v id e d by
inactivated vaccin es is com pletely establish ed in 3-4 w eeks a n d , for
so m e d ise a se s, lasts lo n g er th en th a t p ro d u c e d b y live vaccin es.
In activated vaccin es are o fte n ad m in istere d , w ith a better b o o ste r
effect, after a priming w ith live vaccines.
Poultry diseases
NEWCASTLE DISEASE
Newcastle Disease
N ew castle disease (N D ) is regarded as the m ost im p o rtan t disease
to be con trolled in poultry. T h e im p ortan ce is n ot only d u e to the
rap id sp read an d d ev asta tin g effects on the in fected b ird s, with
flock m ortality rates Lip to 100% , b u t also to the e co n o m ic im p act
that m ight ensue, due to trad in g restrictions an d em bargoes placed
on countries, where outbreaks have occurred. B ecause o f the severe
nature o f the disease, N D is included in the list o f diseases reported
by O ffic e In te r n a tio n a l d es E p iz o o tie s (O IE ), w h ich c o n ta in s
tran sm issib le d iseases w ith a p o te n tial fo r very serio u s an d rapid
spread. M o st m em ber countries enforce statutory con trol m easures
in the event o f outbreaks o f disease. N D is caused by a param yxovirus
se ro g ro u p 1 (A P M V T ) b e lo n g in g to the fam ily Paramyxoviridae,
genus Avulavirus] only one serotype is known, b u t different pathotypes
exist: very virulent (velogenic), m edium virulent (m esogenic), m ildly
virulent (lentogenic), or ap ath ogen ic strains. T h ese latter two types
o f strain s are c o m m o n ly used in the p rep aratio n o f live vaccines.
Epizootiology
O v er 2 5 0 sp ecies o f b ird s have b een re p o rte d to be su scep tib le.
N D viru s is h ighly co n ta g io u s, h orizon tally tran sm itte d th ro u gh
in fected respiratory d isch arge and d ro p p in g s, by either in h alation
or in gestio n . T h e sp re ad a m o n g farm s, even at a long d ista n c e ,
o ccu rs by m o v em e n t o f live b ird s (gam e an d ex otic b ird s, feral
pigeons, com m ercial m arket poultry), m ovem en t o f con tam in ated
poultry- products, people, equipm ent, feed and water and by airborne
spread. Vertical transm ission is very controversial, probably occurring
only very rarely in newly hatched chicks.
Clinical signs and lesions

D ependin g on the signs, lesions and tissue tropism , N D virus strains
have b een d iv id ed in to five g ro u p s or p ath o types: 1) viscerotropic
velogenic: highly v iru le n t d ise a se w ith h igh m ortality an d typical
h aem o rrh ag ic an d n ecrotic g a stro in te stin a l lesion s; 2) neurotropic i
velogenic: resp iratory an d n erv ou s sign s w ith very high m ortality;
3) pneumotropic mesogenic: respiratory and, in som e cases, nervous signs
w ith c o n sid e ra b le m o rtality in y o u n g b ird s b u t low in a d u lts;
10
4) ptieumocropic lentogenic mild or unapparent infection o f the respiratoiy

tract, w ith n o mortality' (u sed a s vaccines); 5 ) naturally apachogenic,
m ostly therm oresistant, used m ore recently as non-stressing vaccines.
In th e field , resp iratory (la b o u re d b re ath in g , rales a n d sneezing),
en teric (green ish d ia r rh o e a ) a n d n e rv o u s sig n s (tw isted n e ck o r
torticollis, leg w eakness o r paralysis, etc.) an d respective lesion s can
all b e present a t the sam e tim e, b u t are n o t strictly p ath ogn om on ic.
T h e d ro p in egg p ro d u c tio n is very rap id , so m e tim e s to zero; eggs
m ay h av e th in a n d d is c o lo u r e d sh e lls o r n o sh e ll; the ovary
ND nervous sign: twisted neck or torticollis
d egen erates an d egg yolk is p re se n t in th e a b d o m in a l cavity, w ith

c o n se q u e n t p e r ito n itis. M o rb id ity m ay re ac h 1 0 0 % ; m ortality
varies, d ep e n d in g on virus virulence an d the residua! im m u n e state
o f b ird s, so m etim e s reach in g o v e r 8 0 % .
Diagnosis
A p resu m p tiv e d ia g n o sis is b a se d o n the ch aracteristic sig n s and
le sio n s a n d is c o n fir m e d by v ir u s iso la tio n a n d id e n tific a tio n
(in oculation o f specim en s, taken from tracheal an d intestinal sw abs
o r tissues, in 9-11 day-old em bryonared eggs, H A an d HI tests with
alla n to ic flu id s). M o re recently m o le cu lar d ia g n o stic te c h n iq u e s
have been d evelop ed , such as R T -P C R an d n u cleotide seq u en cin g.
T h e laboratory asse ssm e n t o f viru s p ath ogen icity is d eterm in ed by
the m ean d eath tim e (M D T ) in cm bryonatcd eggs, the intracerebral
p ath o gen ic index (IC PI) tor one-day-old chicks and th e intravenous
p ath o gen ic in dex (IV Pf) fo r six-week-old S P F ch ick en s (see table).
T h e se an d o th e r m o lecu lar p ro p erties allow d istin c t viral p rofiles
to b e d e v e lo p e d , d istin g u ish in g b etw een av iru le n t an d v iru le n t
isolates. A n tib od ies are d em o n strated u sin g the haem agglutination
in h ibitio n (H I) o r E U S A tests.
Control of disease
T h e objectives are b o th ro prevent su sceptible b ird s from becom ing
infected and to reduce the num ber o f susceptible birds by vaccination.
C o n tro l p o lic ie s a re ap p lie d a t in te rn a tio n a l, n a tio n a l an d farm
level a n d c o n s ist o f th e a p p lic a tio n o f all b io se c u ritv m e asu re s
directed a t preventing th e in tro d u ctio n a n d sp re ad o f virus within
c o u n tries an d areas. T h e e stab lish m e n t o f restrictive m easu res on
th e m ovem en t o f b ird s an d their p ro d u cts, q u aran tin e proced u res
fo r im p ortation o f all kind o f d o m estic o r wild birds, th e stam pingo u t o f ou tb reak s o f d isease, n ation al an d in ternation al surveys and
reports o f disease ou tb reaks - everything is organized an d controlled
by th e H ealth A u th o ritie s u n d er O I E . E xcep t fo r a few co u n tries
N E W C A S T LE DISEASE
(Scan dinavia, N ew Zealand an d Sw itzerland), vaccination is adopted

universally, m o st recently even in A ustralia, particularly in intensive
r e a r in g a r e a s . V a c c in a tio n u s u a lly p r o te c ts b ir d s fr o m th e
con seq u en ces o f th e d isease, interfering with th e sp read o f virulent
v iru s, alth o u g h v iru le n t v iru s re p licatio n a n d s h e d d in g m ay still
occur, albeit at reduced levels. In any event, it well applied, vaccination
represents a n effective b arrier to the sp read o f infection. T w o types
o f vaccin es are u sed , live an d inactivated.
Live vaccines:
ND haemorrhagic lesions in proventriculus
T h e se are p re p are d w ith th e in fected allan to ic
flu id o f e m b ry o n a te d eggs k ep t freeze-dried. T h e stra in s o f viru s

u se d fo r v a c c in e p r o d u c tio n a re d iv id e d in to 3 g r o u p s , fully
ap ath o gen ic, len togen ic an d m esogen ic (sec table).
Mesogenic strains a re o fte n u sed on ly in co u n trie s o r a re a s w here
N D is en d em ic, w ith w id esp read p resen ce o f backyard b ird s an d
n o t very in te n siv e rearin g. T h e y c au se very sev ere p ost-vaccin al
re a c tio n s, so m e tim e s w ith so m e sig n s o f d ise a se ; th e re fo re they
are u se d , w h ere it is p e rm itte d , m o stly fo r re v a c c in a tio n an d
in tram u scu larly , afte r a p rim in g w ith ap ath o g e n ic o r len togen ic
vaccin e. Lentogenic strains genotype II, su ch a s H itc h n e r B 1 a n d L a
S o fa (1CPI 0.2-0.4 L w hich rep licate particu larly in the m u co sa o f
th e resp irato ry trac t an d can in d u c e p ost-vaccinal re a c tio n s and
re sp ira to ry sig n s, p a rtic u la rly in y o u n g c h ic k s a n d in p rim ary
vaccin ation , d e p e n d in g o n th e h ealth status, intercurrent bacterial
in fe c tio n s (Mycoplasma, E. coli) o f th e b ird s a n d e n v iro n m e n tal
c o n d itio n s (a m m o n ia , d u st, etc.); th e La S o ta stra in is th e m o st
stressing an d n o t recom m en ded fo r use by spray as a first vaccination.
Apathogenic strains genotype I (viru s m u ltip lie s well in en te ric an d
ND necrotic lesion or ulcer in intestinal mucosa
respiratory tracts), such as N D V 6 /1 0 , U lste r 2 C , Q u e e n sla n d V 4,

iso lated fro m w aterfow l an d ch icken s, are fully asy m p tom atic with
an 1CP1 <0.15. Even w ith fine sp ray v accin atio n (d ro p lets < 70-80
m icron s), an d a t an y age, th ey in d u c e n o post-vaccinal reaction s;
b e in g rh e rm o re sistan t, they c a n be u sed w ith few er p ro b le m s o f
storage even in h o t se a so n s an d in tropical areas. T h e ap p licatio n
o f live ap ath o g e n ic, len togen ic strain s a s vaccin es can b e d o n e by
in dividu al treatm en t, su ch as n a sa l-o r eye-drop a d m in istra tio n o r
beak -d ip p in g . T h e m ass a p p lic a tio n of v a cc in e s is d o n e via the
d rin k in g w ater, a t c o n c e n tratio n s carefully calcu lated to give each
b ird a su ffic ie n t d o se o f v iru s; a d d itio n o f d rie d sk im m e d m ilk
pow der to fresh d rin k in g w ater (1-2% ) is suggested to prevent viral
inactivation by physical an d chem ical im purities, with no concurrent
use o f disinfectants. B efore vaccination, the birds have ro be deprived
o f w ater fo r 1-2 h o u rs, d e p e n d in g o n the se a so n (see later).
T h e m ass app lication o f live vaccines by spray o r aerosol is also very

w id esp read , b ecau se a large n u m b e r o f b ird s can b e treated in a
very sh o rt rim e. S p ray vaccination can b e perform ed in the hatchery
o r o n the farm . T o achieve the correct d rop let size is very im portant.
A coarse spray o f relatively large particles (> 100 m icrons) d o e s not
p en etrate d eep ly in to th e resp iratory tract o f b ird s an d p ro d u c e s
less o f a reactio n ; this is less condition ing with apathogenic strains
having an ICPI = 0.0. T h e sp ray m eth o d a t one-day-old m ay resu lt
in th e e sta b lis h m e n t o f in fe c tio n w ith v a c c in a l v iru s, d e sp ite
m a ter nally-d e r ived i m m u n ity.
Inactivated vaccines: T h e se
are p rod u ced with allan to ic flu id s
from em bryon ated eggs, infected w ith d ifferen t len togen ic strain s,
such as L a S o ta o r U lster 2 C , rarely with m esogen ic strain s (R oakin
strain ), generally n o lon ger w ith velogen ic strain s, in activated with
fi-p ro p io lacto n e o r fo rm a lin an d m ixed w ith m in eral oil to form
a stable em ulsion. O n e o r m ore ad d ition al viral o r bacterial antigens
m ay b e in corporated in to the sam e em u lsion a s polyvalent vaccines.
T h e v a cc in c is a d m in iste re d by in tra m u sc u la r o r su b c u ta n e o u s
in jection. T h e se vaccines are usually u sed in pullets, on ce o r twice,
b e fo re g o in g in to lay, g iv in g lo n g -lastin g im m u n ity . T h e y have
so m etim es been used in broilers, a t one-day-old, togeth er with live
vaccin e, particu larly in en d em ic are as o f d isease o r in c ase o f very
severe ep id em ics.
Characteristics o f some NDV strains

TEMPERATURE
RESISTANCE
VIRUS STRAIN
PATHOTYPE
GENOTYPE
ICPI
6 /1 0 , Ulster 2 C, V4
A pathogenic
< 0 .1 5
Field isolate
H itchner B l, A splin F,
La Sota
P
Lentogem c
n
II
0.2
Q4
Field isolate
Fie,d ,solate
t.
[j0
A,irK.
H, Muktesvar, K om arov
M esogenic
csugcniL
1!
] 4
Field isolate
Aegg
ttenuated
by
passages
Herts 3 3 /5 6
Velogenic
111
2 .0 0
Field isolate
ORIGIN
13
Avian Pneumoviruses
A v ian p n e u m o v iru se s are m em bers o f the fam ily Paramyxoviridae,
g en u s Metapnewmovirus. In fection s, affectin g prim arily turkeys and
c h ick en s, a n d h ave been term ed turkey rhinotracheitis (T R T ) an d
chicken swollen head syndrome (S H S ) respectively. T h e d ise a se in
ch ick en s w as first rep orted in S o u th A frica in the late 1 9 7 0 s.
It later sp read to E u ro p e, particularly to turkeys a n d ch icken s, first
to Italy a n d th e U .K ., th en elsew here); afterw ard s an d to a lim ited
ex ten t, to th e U S A , Ja p a n , Brazil a n d p ro b a b ly to a n u m b e r o f
o th e r co u n tries. S tu d ie s u sin g m o n o clo n al an tib o d ie s have m ad e
it p o ssib le to d istin g u ish fo u r subty p es o f the virus, referred to as
A , B, C an d D.
Epizootiology
C o u n trie s re p o rtin g iso la tio n s o f the viru s are relatively few, an d
th ese particu larly in turkeys; p resen ce o f in fection h a s frequently
b een b a sed o n serological evidence. C u rre n t evidence suggests th at
the virus is on ly likely to be sh ed for a few days after in fection and
th at n o carrier state exists. S p read o f the in fection occu rs by contact
a n d is a irb o rn e ; m o v e m e n t o f e q u ip m e n t an d p e o p le m ay also
b e im p licated , alth ou gh resistan ce o f th e viru s is nor high.
S p read over greater d istan ce s is ap p aren tly sligh t.

T R T , particularly in y o u n g p o u lts, is characterized by m ore o r less
ev id en t sn eezin g, co u g h in g , nasal d isch arge, sw ollen in frao rb ital
sin uses, con jun ctivitis an d head shaking. In laying b ird s, th ere may
b e a d r o p in egg p r o d u c tio n o f up to 7 0 % , w ith a n in cre ase d
in cid e n c e o f p o o r egg sh ell quality. M o rb id ity is can b e 1 0 0 % ,
m ortality varies greatly, from less than 1% to as high as 3 0 % , m ostly
d epen din g on the severity o f bacterial co-infections, which com plicate
th e viral in fection .
In th e p u re fo rm , th e on ly le sio n is th e p re se n c e o f e x u d a te in
t u r b in a te s a n d in fr a o r b ita l sin u se s; in m a tu re fe m a le s , so m e
a b n o r m a litie s are o b se rv e d in the ov ary a n d o v id u c t, su c h as
regression, fo ld ed shell m em b ran es in the ovidu ct, m issh ap en eggs,
p eriton itis.
JA
W h e n se c o n d a ry b acterial co -in fcctio n s o c c u r (m ostly - coii), a

variety o f gross lesion s have been observed, in clu d in g p n e u m o n ia,
p e r ic a r d itis, a ir s a c c u litis a n d m e n in g itis. S H S in c h ic k e n s is
ch aracterized by th e s a m e resp irato ry sy m p to m s a n d sw ellin g o f
periorbital sinuses, ear and eye discharge, opisth oton u s and torticollis.
U su ally less th an 3 % o f the flock is so affected, although respiratory
sig n s m ay b e w id e sp re a d . In heavy b re e d e rs a re d u c tio n in egg
p ro d u c tio n , o f th e o r d e r o f less th an .5%, m ay occu r. M ortality
rarely ex ceed s 1-2% . T h e o n ly sig n ific a n t le sio n is an exten sive
Poult affected by TRT: sinusitis and conjunctivitis
yellow, g e la tin o u s to p u ru le n t su b c u ta n e o u s o e d e m a o f the h ead

an d , so m etim e , the w attles.
Diagnosis
C lin ical d iagn osis, particularly in turkeys, can on ly be presum ptive,
b ecau se m any o th e r c o n d itio n s, su ch as m y co p lasm osis, influenza
a n d N ew castle d ise a se , c a n sh ow sim ilar sym ptom s.
T h erefo re, o th e r in vestigatio n s, su ch a s viru s iso la tio n a t the first
sig n s o f d isea se , m o le cu lar id e n tificatio n (R T -P C R ) an d serology
(E L ISA , im m unofluorescence) are required tor a definitive diagnosis.
V iru s iso la tio n , n o t so easy to d o , is p e rfo rm e d in e m b ry on ated
eggs, via the yolk-sac, and in em bryo tracheal organ cultures (T O C ).
Control
Live a tten u ate d v a cc in e s are u sed bo th in turkeys a n d ch ick en s.
W h en adm inistered correctly, by coarse spray, these vaccines provide
g o o d p ro tectio n , stim u latin g b o th system ic a n d local im m u n ity in
the resp iratory tract. G o o d cro ss p ro te c tio n a m o n g th e su b ty p es
occu rs. U su ally , turkey b roilers are vaccinated a t 1, 10 an d 4 5 days
o f age; ch ick en s are vaccin ated on ly o n ce , betw een 1 an d 10 days
o f age. In breeders, live vaccines are o fte n follow ed by an injection
o t in activated vaccin e in oily e m u lsio n , tw o w eeks b e fo re th e start
o f laying.
Avian Influenza
A vian in flu en za (A I) is cau sed by a viru s b e lo n g in g to th e fam ily
Orthomixoviridae, g e n u s Influenza vines, serotype A. A l v iru se s have
a w orldw ide distribution. In fection can o ccu r in m any fo rm s, from
asym p tom atic ro respiratory d isease a n d a d ro p in egg p ro d u ctio n ,
to system ic d isease, with a m ortality rate o f u p to 100% . T h erefore,
viral iso la te s are su b d iv id e d in to low (L P ) an d high p a th o g e n ic
(H P); so m e tim e s, stra in s o f so m e su btyp es (to d a te , H 5 an d H 7)
can m u ta te fro m LP to H P. H igh ly P a th o g e n ic A v ian In flu en za
(H PA 1) is in clu d e d in the list o f d ise a se s re p o rted by th e O ffic e
Chickens affected by HPAI before death
In tern ation al d e s E p izooties (O IE ), sim ilarly ro N ew castle D isease.

In flu en za A v iru s is c la ssifie d in sa b types, b a se d o n se ro lo g ic al
r e a c tio n s o f s u r fa c e g ly c o p r o te in s , h a e m a g g lu tin in (If) an d
n e u ra m in id a se (N ). S ix te e n H a n d 9 N an tig e n s h ave s o fa r been
recognized; m any com bin ations o f H an d N antigen in virus subtypes
have been reported in d om estic an d wild birds. A ntigenic variations,
m ore (sh ift) o r less (d rift) w ide, h av e b een re p o rte d in pou ltry,
th o u g h few er th an in m am m als.
Epizootiology
T h e m o st fr e q u e n t so u rc e o f A I v iru s a re w aterfow l, esp ecially
d u ck s, geese , g u lls a n d o th e r w ild b ird s, w hich are c o n sid e re d as
th e n atu ral reserv oirs; in th e se latte r sp ecies, A I v iru s in fe c tio n
usually cau ses sp orad ic o r n o disease. A I virus has, in the p ast been,
sporadically isolated from dom estic integrated com m ercial poultry,
m o st frequently turkeys an d , slightly less o fte n , ch icken s an d oth er
d om estic birds. A n im als from th e live p ou ltry m arket (LPM ) system
a n d backyard flocks have so m e o f the h igh est in fection rates.
However, ou tb reaks o r actual epizootics in com m ercial poultry have
beco m e m ore a n d m o re freq u en t in the last 2 0 years, particularly
recently, w orldw ide. W hen A I infections d o occur, they, som etim es,
sp read rapidly th rou gh the integrated poultry system , fro m farm to
farm , resu ltin g in ep izootics o f L P o r H P A I. In so m e d e v e lo p in g
co u n trie s o r in so m e are as w h ere th e L P M system is w id esp read ,
A I h a s often becom e en d em ic.
A I viru s is excreted by th e resp iratory a n d en teric routes, s o it is
transm itted by d irect con tact betw een infected an d susceptible birds
16
o r by in direct con tact through aerosol d ro p lets o r ex p osu re to virus

con tam in ated fom ites, eq u ip m en t and p eople. S o u rces o f in fection
lo r initial in tro d u ctio n o f A I virus in to com m ercial poultry flocks
in clude o th er d o m estic an d con fin ed poultry, m igratory waterfowl,
p e t b ir d s o r, in s o m e c a se s, even d o m e s tic p ig s. In tra sp e c ie s
tran sm issio n is fre q u e n t an d easy; in te rsp e cie s tra n sm issio n may
o c c u r, b e in g e v e r le ss fr e q u e n t a s th e d iffe re n c e s b etw een the
p h ylogen etic classes o f th e an im als in crease; b u t s o m e ex cep tion s
have o ccu rred , alth o u g h a t p re se n t ro a relatively lim ited extent:
Turkeys affected by HPAI
H IN 1 from sw ine to turkey in the U S A an d , move recently, H 5N 1

an d H 7 N 7 fro m p o u ltry to h u m a n s, resp ectiv ely in A sia a n d in
T h e N etherlands. A lthough the transm ission o f infection com m only
o c c u rs h orizo n tally, ev id e n c e o f vertical tra n sm issio n is lackin g;
how ever the virus h as b e e n iso lated fro m th e in tern al c o n te n ts o f
the eggs d u rin g n atu ral o u tb re ak s o f d isease.

C lin ic a l sig n s a re ex tre m ely v a ria b le , d e p e n d in g o n th e viru s
p athotype an d other factors, in cluding h ost species, age, concurrent
in lection s an d en viron m en tal c o n d itio n s.
LPA1 v iru s in fe c tio n in w ild b ird s u su ally p r o d u c e s n o sig n s.
H ow ever, in d o m e stic p ou ltry it n orm ally p ro d u c e s m ild to severe
resp iratory sy m p to m s, su ch as cou g h in g , sn eezin g, rales, to g eth er
w ith v a ria b le m alaise, a n o re x ia a n d d ia rrh o e a ; in b re e d e rs an d
layers, a d ecrease in egg p ro d u c tio n (varying fro m 10 to 8 0 % ) is
a lso p o s s ib le . S o m e tim e s h igh m o r b id ity , b u t lo w m o rta lity
(less th a n 3-5% ), is th e ru le, u n le ss c o m p lic a te d by se c o n d a ry
in fectio n s o r it the d isease o ccu rs in very y o u n g birds.
T h e m o st fre q u e n t lesio n s in th e resp irato ry tract are: catarrhalfibrinous to p u ru len t sinusitis, tracheitis, bron ch op n eu m on ia, m ore
o r less severe airsaccu litis, also d e p e n d in g o n sec o n d ary bactcrial
in fection s; catarrh al to fib rin o u s en teritis an d o v id u ctitis m ay also
be observed.
H P A I v iru s in fe c tio n in w ild b ird s an d d o m e stic d u c k s u su ally
p ro d u c es few o r n o clin ical sign s; however, a few c ase s have been
rep o rted w ith m o d e ra te to h igh m ortality . In d o m e stic poultry,
such as turkeys, ch ick en s, gu in ea fowl an d q u ail, the clin ical signs
vary, d epen d in g o n the degree o f dam age induced by viral replication
in to th e sp ecific o rg a n s an d card io v asc u lar an d n erv o u s system s.
In m o st cases, the disease is fulm inating, in the absence o f an y clear
sym ptom ; if b ird s survive fo r a few days, so m e o f th em m ay exh ibit
re sp ira to ry a n d n e r v o u s d is o r d e r s . R e sp ir a to r y sig n s a re less
p ro m in e n t th an with LPA1. A p recip ito u s d ro p in egg p ro d u ctio n

is the rule. M o rb id ity an d m ortality is very' high (50-90 to 100% ).
E d em ato u s, h aem orrh agic an d n ecrotic lesion s in visceral organ s,
often in clu d in g the p an creas an d non-feathered sk in , are observed,
h ae m o rrh a g e s a re p ro m in e n t in th e e p ic a rd iu m , p ro v e n tric u lu s
a n d Payers p atch es. In s u d d e n d eath , n o g ro ss le sio n s b u t on ly a
gen eral severe co n g estio n m ay b e observed.
Diagnosis
Chickens broiler affected by HPAI
A p resu m p tiv e d ia g n o sis o f A I is b a se d o n e p id e m io lo g ic a l an d

clinical d ata, w hich, how ever, are n o t p ath o gn o m o n ic. A definitive
d iag n o sis is estab lish ed by d irect d etectio n o f an tigen s o r D N A o f
th e virus in variou s sp ecim e n s (tracheal a n d clo aca! sw abs, tissues,
etc.), and by isolation and identification o f the virus in em bryonated
eggs o r by R T -P C R , follow ed by d e te ctio n o f an tib o d ie s (E L IS A ,
H I, A G P , V N ).
Control
M e th o d s fo r the c o n tro l o f A I virus in fection arc m ainly based on
k n o w led ge o f how th e viru s is in tro d u c e d in to a farm o r an area
a n d how it can sp re a d . T h e re se rv o ir o f the v iru s in w ild b ird s
sh o u ld b e c o n sid e r e d a m a jo r so u rc e o f p rim ary in fe c tio n fo r
com m ercial birds, particularly free-range birds. T h e strict application
o f biosecurity- m easu res is th e first line o f d efen ce, p reven tin g n o t
on ly d irect, b u t also in d irect c o n tact o f c o n ta m in ate d eq u ip m e n t,
fo o tw ear, c lo th in g , veh icles o r p e o p le w ith in fe c tio u s m aterials
(excreta).
C o n ta m in a te d p o u ltry m an u re ap p e ars to b e a m o st likely so u rce
o f v iru s sp re a d betw een flock s o r farm s, even a t lo n g d ista n c e s.
C on siderable difficulties are som etim es encountered with application
o f all b io se c u rity m e asu re s, p articu larly in a re a s w ith very high
c o n c e n tratio n s o f farm s. W h en in fe ctio n by su b ty p e H 5 an d H 7
(the on ly su b ty p es w hich have so far sh o w n a cap acity to con vert
from LPA I ro H PA I) is detected, the resp o n se m u st be p ro m p t and
c o m p le te , even re so rtin g to im m e d ia te c u llin g o r d e p o p u la tio n .
A s regard s p rev en tio n by v a ccin atio n , in activ ated vaccin es in oily
e m u lsio n h av e b een u se d , p articu larly in c h ic k e n s a n d turkeys.
T h e ir effectiveness in preventing sy m p to m s and m ortality h as been
w ell d o c u m e n te d . H ow ever, d iffic u lties in p re p arin g a vaccin e in
advance arise, becau se there are 16 d ifferen t H subtypes o f AI virus.
W h en an o u tb re ak o f A I occu rs an d the virus su btype is identified,
v a cc in atio n m ay b e a u se fu l to o l, p articu larly in a re a s w ith high
18
p oultry p o p u la tio n d en sities, b u t the p re p aratio n o f a v accin e h as

to b e q u ite rap id. V accin atio n sh o u ld be con sid ered a valid op tio n
to c o n tr o l th e sp r e a d o f in fe c tio n , re m a rk a b ly r e d u c in g the
su sc e p tib ility o f b ird s to th e in fe c tio n an d th e a m o u n t o f viral
sh e d d in g in the environ m ent. T h e con trolled u se o f vaccines, ever
in cases o f LPAI H 5 an d H 7 ou tb reak s m igh t redu ce the possibility
o f H P A I v iral em e rg e n c ie s; th e ir u se c o n tin u e s to b e d e b a te d ,
particularly in E u ro p e a n d N o rth A m erica.
H eterologous vaccine h as also been used for the N antigen, applying
the D IV A (differentiating infected from vaccinated anim als) strategy,
b a sed o n se ro lo g ic a l te sts to h igh lig h t d iffe re n t sp e c e fic anti-N
antibodies. T h e trend seem s to b e in this direction, also considering
the recent exam p les o f resu lts o b tain e d in certain countries.
T h e ju d ic io u s u se o f v a c c in e , m a k in g it p o s s ib le to re d u c e
tran sm issio n an d sp read o f infection, an d the susceptibility o f birds
to the virus, w ou ld favour eradication o f the disease, also preventing
it from b eco m in g en d em ic.
Layinq curve in an outbreak o f LPAI (subtype H7N1) in a farm o f 4 houses

with layers o f different ages
Infectious Bronchitis
Infectious bronch itis (IB) is an acute, highly con tagiou s viral disease
o f poultry, cau sed by a viru s classified in th e fam ily Coronaviridae,
g en u s CoroncR'irus, w hich particu larly affects th e resp iratory tract,
b u t also th e k id n e y s a n d re p ro d u c tiv e trac t. T h e o c c u rre n c e o f
m u ltip le sero ty p es (to d ate over 6 0 d ifferen t sero ty p es have been
rep orted all over the w orld) seem s to co m p licate the prevention o f
th e d isease by vaccination.
Epizootiology
Trachea:
a) normal
b) affected by IB
IB (1BV) c a u s e s c o n s id e r a b le d a m a g e to th e p o u ltry in d u stry

w o rld w id e. C h ic k e n s a n d , ro a lesser ex te n t, p h e a sa n ts a re the
prim ary n atu ral h o sts. A ll ag es are su sc e p tib le , b u r th e d ise a se is
m ore severe in y ou n g ch icks.
IB virus (IBV ) sp re ad s rapidly in a flo ck o r in a farm , w ithin 24-48
h o u rs; th e viru s is excreted by c o u g h in g an d sn eezin g fo r a b o u t 10
days, b u t rhe in fection can persist to r w eeks o r even m on th s in the
k idn eys a n d gut, bein g sh ed by d ro p p in g s.
W ith rhe ex cep tio n o f th e M assachusetts serotype, p re se n t all over
th e w o rld , o th e r se ro ty p e s are m o re lo c a te d in d iffe r e n t areas,
c o u n trie s o r c o n tin e n ts (Connecticut, Gray, A Z- 2 3 /7 4 , A rkansas
DPI, U K -4 /9 1 , A Z 2 7 / 9 8 or h-02, A Z -4 0 /0 5 or Q X lik e etc.).

T h e c h arac te ristic re sp ira to ry sig n s in y o u n g ch ick s are g asp in g ,
c o u g h in g , sn eezin g , n asa l d isch arg e, trach eal rales, o c c asio n ally
sw ollen sin u ses, to g eth er w ith m ore o r less severe d e p r e ssio n an d
reduced w eight gain. In b ird s old er th an 6 weeks an d in ad u lt birds,
the sign s a re sim ilar, b u t less severe.
In fec tio n in ch ick s, a few d ays o ld , is rep orted to p ro d u c e so m e
p e r m a n e n t d a m a g e to th e o v id u c t, w ith re d u c e d fu tu r e egg
p ro d u c tio n a n d a p p e aran ce o f false layers : in the fie ld , th is is
Kidney lesions caused by nephropathogenk IBV
n o rm ally prevented by th e p resen ce o f g o o d levels o f m aternallyderived an tib o d ies.

In laying flocks, a decline in egg p ro d u ctio n an d quality is observed,
in a d d itio n to m o re o r less ev id en t respiratory sy m pto m s.
T h e severity o f the d ro p in egg p rod u ctio n varies greatly, from slight
20
to over 5 0 % , varying w ith the p eriod of lay, causative virus strain

an d in com plete vaccinal im m unity. F o u r to eight weeks may elapse
before egg prod u ctio n returns to the pre-infection level, but in m ost
cases this is never attain ed. D isco lo u red, rough- or soft-shelled and
m issh ap en eggs are p rod u ced , hatchability is reduced.
C hicken s, particularly broilers, affected by neph ropath ogenic strains
of IBV, in a d d itio n to the m ore or less severe typical respiratory
sy m p to m s, sh ow sign s of severe d ep re ssio n , w et d ro p p in g s w ith
urates, in creased w ater intake, d eh yd ratation .
M orbidity1 m ay be very h igh , m ortality is variab le, d e p e n d in g on
v iru le n c e of the viru s strain , im m u n ity sta tu s, cold stress an d
Characteristics of eggs laid from hens affected
by IB:
a) normal
b) different degree of alteration
secon d ary b acte rial in fectio n s (tmcopl/ismfls, Escherichia coli, etc.),

w hich c o m p lic a te th e co u rse of illn ess, p articu larly in b ro ile rs
(chronic respiratory disease).
A t necropsy, in fected ch ickens show serous-catarrh al or fib rin ou s
exu d ates in nasal p assages an d sin u ses, b u t m ostly in the trachea
and air-sacs. A caseous plug may be found in the tracheal bifurcation
an d bronchi in you n g chicks. In n e p h ro p ath ic in fection s, sw ollen
and pale kidneys, due to interstitial nephritis and d isten ded ureters
with urates are present; in layers, after the acute phase, urolith iasis
m ay occur.
Diagnosis
A presum ptive diagn osis of IB is based on the rapid onset o f clinical
sign s an d on the lesion s, w hich, how ever, are n o t very sp ecific.
Lt is confirm ed by the detection o f IBV antigen (im m unofluorescence
assay), iso latio n and even tu al typing o f the viru s (in o c u la tio n in
em b ryon ated eggs or trach eal organ cu ltures) an d R T -P C R from
affected tissues or em bryo fluids. Later, serological tests are applied
to detect group-specific an tib o d ies (A G D an d E L IS A tests) or typespecific an tib o d ie s (H I, V N ).
Embryo lesion in SPF eggs inoculated vsith IBV:
dwarfing, curling, persistence of mesonephros
T h e w id e a n tig e n ic v a r ia tio n s of IB V ad d c o m p le x ity to the

co n d u ctio n o f tests an d the analysis o f results. C ross-reaction s are
m ore evident w hen sera are collected after field infection in broilers,
b u t m ain ly in layers, w h ich m ay have receiv ed o n e o r m ore
vaccin ation s or in fection s.
Control
B iosecu rity m easu res ag ain st IB are very' d ifficu lt to apply, d u e to
the high density' o f farm s in certain areas an d w hen m ultiple ages
are p resen t in the sam e farm . A logical con seq u en ce is the n eed to
resort to vaccin ation to preven t losses of p ro d u ctio n .
21
B o th live a tte n u a te d an d in ac tiv ate d v a c c in e s are u se d in IB

im m un ization . In b roilers, on e or two vaccin ation s are p erform ed
with a live vaccine, m ostly at 1 and 12-20 days o f age. In b reed ers
an d layers, a com b in atio n o f live vaccines, adm inistered d u rin g the
pullet (2-3 tim es) an d laying p e rio d s (every 3 m on th s) an d on e or
two in je ctio n s o f in activ ated v accin e (p refe rab ly p re p are d with
2 o r 3 d iffere n t serotypes) b efo re en terin g the laying p e rio d , are
recom m en ded .
IB V strains used for live vaccine p rep aration have been atten u ated
by serial p assages in em bryonated eggs, avoid in g excessive nu m ber
o f passages to prevent red u ctio n o f im m unogenicity.
V accin e prepared w ith M assach u setts serotype con tin u es to be the
m o st co m m o n ly used vaccine in the w orld, also b e cau se viru ses
iso la te d in m o st c o u n trie s b e lo n g to th is sero ty p e . T h e m o st
w id e sp re a d v a c c in a l strain is n am e d H 1 2 0 ; it b e lo n g s to the
M a ssach u setts serotype, b u t oth e r strain s are also p re se n t o n the
m arket. T h is serotype provides fairly good cross-protection, which,
however, d oes n ot cover com pletely all other serotypes o r varian ts
o f the virus. O ther serotypes are used in live attenuated or inactivated
vaccines w orldw ide. U n lik e N D an d IB D , m aternal an tib o d ie s d o
n o t interfere greatly with vaccin ation p erform ed at on e day o f age.
IN F E C T IO U S LA R Y N G O TR A C H E 1TI5
Infectious Laryngotracheitis
In fectiou s Laryngotrach eitis (IL T ) is cau sed by a virus classified in
the fam ily Herpesviridae, genus Ct-herpesi'irus, w hich replicates in the
respiratory tract.
Epizootiology
C h ic k e n s are the p rim ary n atu ral host., p articu larly ad u lt bird s.
N u m ero u s cases of IL T have also been reported in ph easan ts; other
b ird s a p p e a r to be refractory. T h e n atu ral rou te o f entry fo r the
vim s is through the upper respiratory and ocular tract. T ran sm ission
o f the d isease occu rs by d irect c o n tact with in fected bird s, by the
I airb orn e rou te or by co n tam in ated e q u ip m e n t an d people.
) Clinical signs and lesions

IL T is usually ch aracterized by acute respiratory symptoms-, nasal
I discharge, coughing, gasping, tracheal rales, expectoration o f bloody
m ucus to casts, due to fibrinous-haem orrhagic tracheitis. M ild form s
o f disease have often been observed, characterized by conjunctivitis,
I sw elling o f in fraorb ital sin u ses an d n asal discharge.
; T h e course o f in fection varies with the severity o f the lesion s: the
! m o st ch aracteristic in acute form s is the h em orrh agic-d ip h th eric
exudate, w hich can extend to the entire length o f trachea.
! I L T in d u c e s a m o re o r less se v e re d r o p in egg p r o d u c tio n
(10 to 60% ), d e p e n d in g on the severity o f disease, for a p erio d o f
3-5 weeks.
i In acute form s, m orbidity is very high; m ortality, how ever, is very
v ariab le, from 5 to 6 0 % , b u t u su ally w ith in the 10-1.5% range.
In m ild enzootic form s, m ortality is rath er low, less th an 2 % .
I Diagnosis
In cases o f severe sign s and lesion s, w ith e x p ectoration o f b lo o d y
m u cu s an d high m ortality, the d ia g n o sis is reliab le. O th erw ise,
laboratory diagnosis procedures are advisable, including the presence
o f in tran u c le ar in clu sio n b o d ie s in tissu es stain ed w ith G iem sa ,
iso latio n o f virus in em bryon ated eggs and P C R .
24
P ou ltry D isease M anual
Control
B esid es the ap p lic a tio n o f a p p ro p ria te bio secu rity m easu res, the
c o n tro l o f the d ise a se in g eo g rap h ic areas, w here it is en d e m ic o r
freq u en t, is based o n th e u se o f atten u ated vaccines. A tte n u atio n
h a s b e e n o b ta in e d by s e r ia l p a ssa g e s o n tissu e c u ltu r e s o r in
em bryonated eggs. T h e m o st efficacio u s rou te o f ad m in istratio n is
by eyc-drop. W h e n the n u m b e r o f b ird s to vaccin ate is very high,
ir can b e advisable to ad m in ister the vaccine via the d rin k in g water,
preferab ly a t tw o su ccessive tim e s a n d ar 1 f - 2 d o se s p er bird , so
Characteristic attitude o f chicken affected by ILT,
during breathing
th at all c h ic k e n s can c o n su m e a n a d e q u a te a m o u n t o f th e virus.

A dd itio n o f skim m ed m ilk 1% to fresh w ater is advisable. Som etim es
adverse post-vaccinal reactions m ay occur, w h en the environ m ental
c o n d itio n s are su b o p tim a l (dust, am m o n ia, cold) o r adm inistration
o f the vaccin e is in co rrect. If a c o n sid e rab le n u m b e r o f ch ick en s
d o n o t receive the vaccin ation , bird-to-bird p assage m igh t resu lt in
reversion o f the viru s to greater virulence.
U su ally , on ly p u lle ts d e stin e d ro b e c o m e layers o r b re e d e rs are
v accin ated ag a in st ILT, w ith a sin g le a d m in istra tio n a t 5-8 w eeks
o r twice, at 3 4 and 16-18 weeks o f age, in high risk farm s. Som etim es,
m ostly in e n d e m ic areas, th e v accin atio n o f b ro ile rs m ay also be
required , particularly w hen flocks are close to o u tb reak s o f disease
or th e b ird s are grow ing over. In su ch cases, broilers are vaccinated
at 14-16 d ays o f age by eye-drop o r, m ore o fte n , via the d rin k in g
w ater, en su rin g th e vaccin e is correctly ad m in istered .
FO W L POX
Fowl Pox
Fowl p ox
(FP)
is a c o m m o n d isease o f p ou ltry (ch ick en s, turkeys,
pheasants, etc.), as well as o f pets an d wild birds, caused by num erous

species o f the g e n u s Avipox virus, fam ily Poxviridae. D ifferen t species
are m ore o r less d istin g u ish a b le fro m each o th e r, b lit with varying
d egrees o f cross-relationsh ip.
Epizootiology
Poxvirus infects b ird s a t any age, b u t m ostly after 8-10 w eeks o f age.
It survives fo r a lon g tim e in the sc ab s an d d ried p u stu le s a n d it is
Pox lesion on eyelids o f a layer
tran sm itted by d irect o r in d ire c t c o n ta c t o r by b itin g m o sq u ito e s

o r m ites; in th e latter case, it can also affect nearby flocks. In so m e
are a s, th e d ise a se is se a so n a l (w et-hot w eath er a n d p resen c e o f
insects).

F P m ay o c c u r in tw o fo rm s, c u ta n e o u s o r d ip h th e ric , o r b o th .
T h e cu tan eo u s form is characterized by the ap p earan ce o f n o d u lar
o r w art-like lesio n s o n th e c o m b , w attles, eyelid s, legs a n d oth er
non-feathered areas o f the skin , yellow to brow n in colou r.
T h e d ip h th e ric fo rm , o r w et p o x , o c c u rs in th e m u c o sa o f the
m o u th a n d o e so p h a g u s a n d , m ain ly, o f th e larynx a n d trach ea,
w ith th e a p p e aran ce o f n o d u le s o r p atch es, w h ich in crease in size
a n d o fte n c o ale sc e to fo rm yellow , cheesy, n e cro tic, d ip h th e ric
m em b ran es o r plugs; in such cases, the bird can even ch oke. If rhe
sc a b s o r d ip h th e r ic m e m b r a n e s a r e re m o v e d , a g r a n u la t in g
h em o rrh agic tissu e, with e ro sio n s, is observed.
M o rb id ity a n d c o u rse o f th e d ise a se are variable, also d e p e n d in g
o n viru s viru len ce, hygiene an d clim ate. T h e cffects o f p o x usually
involve em aciation , p o o r w eight gain an d m ore o r less redu ced egg
p ro d u ctio n . T h e course is 3-4 o r m ore w eeks. Mortality- also varies,
Pox lesion on legs o f chicken
from 1-2%, w hen m ild cu tan e o u s le sio n s are p resen t, to over 30 %

w hen d ip h th e ric fo rm s are prevalent.
Diagnosis
W art-like le sio n s o f the c o m b , w attles an d eyelids a n d d ip h th eric
lesio n s in rhe m o u th , larynx an d trach eal m u co sa stron gly su ggest
26
the p resen ce o f this d isease. A definitive d iag n o sis can be m ad e by

a histological exam in atio n .
D ifferential d iagn osis is som etim es necessary w ith laryngotracheitis;
a histological e x am in a tio n can be definitive.
Control
S o m e tim e s th e rem oval o f w arts a n d d ip h th e ric m e m b ran e s an d
th e tre a tm e n t o f lo c al le sio n s w ith d isin fe c ta n ts a re p erfo rm ed .
However, the con trol o f FP is based o n im m unization o f birds with
Pox lesion on the beak o f a pigeon
atten uated vaccin es before th e seaso n , w hen the d isease is likely to

occur; in m u ltiaged farm s an d in tropical clim ates, th e vaccin ation
m ay b e p erfo rm ed a t any tim e o f the year. Even w hen a n o u tb re ak
is a t a n early stage, it is advisable to vaccinate the flock im m ediately,
w hich very o fte n p reven ts the d ise a se from sp read in g an y further.
T w o types o f vaccines can he used, mainly, using FP virus attenuated
by n u m e ro u s p assag e s in e m b ry on ated eggs or o n tissu e cultures,
or non-attenuated pigeon pox-virus. B o th vaccinal viruses are grow n
in em b ry on ated eggs o r o n tissue-cultures and freeze-dried.
T h e vaccine is ap p lied by th e wing-web m eth od , u sin g a stick with
tw o gro o v ed n eed les d ip p e d in vaccine so lu tio n . In b reed ers and
layers, a sin gle vaccin ation is given benveen 4 a n d 14 w eeks o f age;
norm ally b ro ilers arc n o t vaccinated, b u t in so m e se a so n s o r areas
it can b e necessary to preserve the integrity o f the skin.
T u rk eys m ay be vaccinated by the wing-web m eth od , b u t it is m ore
ad v isab le ro d o s o in th e m id d le o f the th igh , by sc arific atio n at
8-12 w eeks o f age; b reed ers sh o u ld b e revaccin ated befo re laying.
T h e p resen c e o f lo cal sw elling a t the in je ctio n site, 8 d ay s after
vaccin ation , sh ow s th at b ird s have b een correctly im m unized.
27
Infectious Bursal Disease

In fec tio u s b u rsa l d ise a se (IB D ), also k n o w n as G u m b o r o d isease,
is an acute, highly contagious viral infection o f young ( 3 /6 - week-old)
ch ick en s, c au se d by a m e m b e r o f th e fam ily B irnaviridae g e n u s
Avybirnovirus, serotype 1. It p rim arily affects th e ly m p h oid tissues:
th e m o st affe c te d o rg a n is th e b u rsa o f F a b ric iu s (B F ), b u t the
rhynuts, sp le en an d o th e r ly m p h oid tissu e s are also involved.
B-lymphocytes are the prim ary target cells. T h e virus (IB D V ) is very
s t a b le in th e e n v ir o n m e n t a n d c a n su rv iv e f o r m o n th s in
c o n ta m in ate d farm s.
Bursa of Fabricius in IBD: diffuse mucosal
haemorrhages
U n til 1987, the strains o f IB D V w ere o f m od erate virulence, usually

c au sin g less th an 5-10% sp ecific m ortality, b u t well co n tro lled by
m ild vaccin es. In rhe late 1 9 8 0 s, v accin atio n failu res b e g an to be
rep orted in d iffe re n t p arts o f the w orld. In N orrh -A m erica it was
d em o n strate d th at the new iso lates h ad been affected by an tigen ic
d r if t / s h i f t (v a ria n ts), w h e re a s in E u r o p e a n d , la te r, o n o th e r
C o n tin e n ts, th e occu rren ce an d sp re ad o f very viru len t stra in s o f
virus (w IB D V ) were reported. T h ese latter strains, even if antigenically
closely related to rhe classic v iru se s (only p u n tifo rm m u ta tio n s),
p ro d u c e m o re severe sy m p to m s a n d le sio n s in ly m p h oid tissu es,
p articu larly in th e th ym u s, w ith very high m ortality , so m e tim e s
over 3 0 % , o cc u rrin g over a p e rio d o f 2-6 days. M ild vaccin es were
sh ow ed to b e a lm o st in effective ag a in st th ese w I B D V , th erefo re
less atten uated an d m ore invasive vaccinal strain s becam e essential.
Epizootiology
T h e chicken is considered rhe only species in w hich natural infection
o ccu rs, b u t a n tib o d ie s have b e e n a ls o d e te cte d in o th e r sp ecics.
A ll b re e d s are a ffe c te d , w ith m o re o r le ss sev ere re a c tio n s an d
Bursa o f Fabricius in IBD: m l! oedema and
catarrhal-haemorrhagic lesions in mucosa
m ortality (genetic in flu en ce). T h e age o f greatest su sce p tib ility to

th e d is e a s e is b e tw e e n 3 a n d 6 w e e k s. IB D V ' is tr a n s m itte d
h orizon tally directly o r in directly, b u t, ap p aren tly , n o t vertically.

T h e in cu b atio n p e rio d o f the d isease is very sh o r t an d the clinical
sig n s o c c u r w irhin 2-3 d ay s o f e x p o su re. C h ic k s, p ick in g initially
ar th eir ow n v en ts, sh o w a n o re x ia , d e p r e ssio n , ru ffled feath ers,
28
w h itish a n d w atery d ia rrh o e a , tre m b lin g a n d severe p ro stra tio n ;

d e a th o c c u rs in a few h ou rs. M o rbidity is very h igh , even clo se to
8 0 -1 0 0 % . M ortality is very v ariab le, b u t it can b e a s h igh as 10 to
7 0 % , d epen d in g on the degree o f pathogenicity o f the virus involved
an d o n en v iron m en tal c o n d itio n s.
In birds which su ccu m b to in fection , d eh yd ratation , h aem orrh ages
in the pectoral a n d thigh m u sclcs a n d , o fte n , in the m u co sa o f the
pro v en tricu lu s, can be d etected . T h e B P is sw ollen, g elatin o u s to
h aem o rrh ag ic; in c ase s o f w lB D V , a g re ate r d e c re ase in thym us
Bursa o f Fabricius in IBD: different appearance
o f more or less severe lesions
si;e a n d m o re severe lesion s in o th e r lym p h oid o rg a n s a rc fo u n d .

A fte r recovery, in acu te fo rm s, in sub-clinical in fection a n d w hen
v a r ia n ts are p r e s e n t, th e B F b e c o m e s m o re o r le ss a tr o p h ic .
T h a t resu lts in a v a riab le im m u n o s u p p r e s s io n , w ith e n h a n c e d
susceptibility to o th e r viral an d bacterial in fection s an d co n se q u e n t
p o o r p e rfo rm a n c e , w ith lo w e r w eig h t g ain a n d a h ig h e r feed
conversion ratio.
Diagnosis
A cu te o u tb re a k s o f IB D a re easily recognized an d a p resu m p tive
d ia g n o sis can b e read ily m a d e (see g ro ss le sio n s); in su b c lin ic a l
c a se s, h isto lo g ic a l le sio n s m ay g iv e in d ic a tio n s, to g e th e r w ith
im m u n o flu o re sc e n c e o r v iru s iso la tio n o n th e c h o rio a lla n to ic
m em b ran e (C A M ) o f 9-11 day-old em bryos, o n B F cu ltu res o r on
BG M -70 cell-1 ines. M ore recently, R T-PC R and m olecular sequencing
have been introduced in the diagnosis. T h e m ost com m o n serological
m eth od fo r an tib o d y evo lu tio n is the E L IS A test.
Control
Hygiene an d disinfection are essential m eans to reduce the infection
rate, h u t generally they are n o t su fficien t, also b ecau se th e viru s is
very resistant to d isin fectan ts. H ow ever, im m unization o f chickens,
b ro ile rs a n d p u lle ts w ith live v a cc in e s a t 2-3 w eeks o f age is the
m ain m e th o d to c o n tro l th e d ise a se . B o o s te r im m u n iz a tio n o f
b r e e d e r flo c k s w ith in a c tiv a te d v a c c in e s in o ily e m u ls io n h as
Haemorrhagic lesions on proventriculus
mucosa in IBD
som etim es proved to b e im portant, in ord er to extend the m aternal

im m unity o f th e progeny to 3-4 w eeks. H ow ever, the p rotection in
broilers d o es n o t extend till slaugh ter, s o they need to be vaccinated
w hen m atern al an tib o d ie s are redu ced (2-3 weeks).
M ild live vaccin es w ere satisfacto rily u sed until th e em ergency o f
virus varian ts in N o rth A m erica an d w lB D V o n o th e r C o n tin e n ts,
w hen the classic prop h y lactic m easu res w ere called in to q u e stio n .
T o c o n tr o l IB D V v a ria n ts, it b e c a m e n e ce ssary ro u se sp e c ific
29
atte n u ate d strain s, to g eth er w ith the classic o n es. L ess atte n u ate d
a n d m o re in vasive v a c c in e s w ere in tro d u c e d in th e c o n tr o l o f
w I B D V . D e p e n d in g o n th eir ab ility to in d u c e im m u n ity in the
p resen ce o f m atern al an tib o d ie s an d th eir residu al pathogen icity,
these vaccines have been classified as in term ediate , intermediatep lu s o r h o t types. A r the m o m e n t, th e first two types a re the
m o st u se d al! over th e w orld, w h ereas th e last is cu rren tly b an n ed
in m any c o u n trie s, d u e to the severe lesion s it cau se s to lym phoid
tissues. It h as been fo u n d th at m arketed vaccines, particularly those
Haemorrhages on the leg muscles in IBD
d e fin e d a s in term e d iate -p lu s , p re se n t very' v a riab le d e g re e s o f

r e sid u a l p a th o g e n ic ity o r in v asiv e n e ss; fo r th is r e a s o n , w id er
investigations an d stan d ard isation sh o u ld be recom m en dable, since
rhe use o f so m e o f th e m is so m e tim e s asso ciated with clear injury
to, an d atroph y o f the B F a n d , probably, o f other lym phoid tissues,
togeth er w ith im m u n o d ep ressio n .
T h e m ain p ro b lem e n c o u n te re d in the c h o ice o f the p ro p e r tim e
fo r vaccination is th at m aternal an tib o d ies interfere with replication
o f v a c c in a l v iru s in ly m p h o id tissu e s. A lth o u g h rhe p a re n ts are
b o o ste d w ith in activated vaccine, th e m aternal an tib o d ie s provid e
a g o o d level o f p r o te c tio n o n ly fo r th e first 3 o r, so m e tim e s,
4 w eeks, then the b ird s b eco m e fully su scep tible.
T h e key fo r efficacio u s p rotection ag ain st IB D is to vaccinate early
en o u g h to p rev en t th e e sta b lish m e n t o f w ild viru s in fec tio n , b u t
late en o u gh to allow the m aternal an tib o d ies to decline sufficiently,
so as n o t to interfere w ith the vaccinal virus. H ow ever, a universal
v a cc in atio n p ro g ram m e c a n n o t be o ffered . T h e b e st m o m e n t fo r
vaccination d ep e n d s o n the rate o f decrease o f m aternal antibodies,
w hich, in its tu rn , is linked to the grow th rare o f th e b ird s, m ore
rapid in broilers th an in light pullets. D ifferent m ethods to determ ine
th e p ro p e r age o f v a cc in atio n h av e b e e n d e v e lo p e d (D a v e n te rs
fo rm u la , cen tral v accin atio n d a te , etc.), b a se d o n an tib o d y levels
ar 1-3 days o f age, b u t it is n o t always p o ssib le to exactly d eterm in e
the p ro p e r tim e, either b ecau se an tib o d y tirres in a flock o f chicks
are h eterogen eou s o r w hen facilities fo r serological co n trols are n o t
a v a ila b le . H o w e v e r, v a r io u s e x p e r ie n c e s (fa r m p a s t h isto ry ,
epidem iological data) can help to im prove the control o f the disease.
In m an y a re a s, th e u se o f o n e v a c c in a tio n a t 18 d ay s o r tw o
vaccinations a t 15 an d 22 days w ith in term ediate vaccine resulted
in g o o d c o n tro l o f th e d isease. In o th e r areas, su c h a s th e tro pics
an d in so m e p eriods o f the year, o r in the p resen ce o f very virulent
wild strain s, the m ore o r less tem po rary u se o f in term ed iate plu s"
vaccine at 12 to 16 days o f age can a t least be tem porarily suggested.
30
A c t iv e I B D V a n t i b o d y d e v e l o p m e n t a f t e r o n e a d m i n i s t r a t i o n o f
i n t e r m e d i a t e o r i n t e r m e d i a t e - p l u s v a c c in e a t 1 8 d a y s o f a g e :
w it h le s s a t t e n u a t e d v a c c in e a n t i b o d y p r o d u c t io n b e g in s a w e e k
e a r li e r
NEOPLASTIC DISEASES - MAREK'S DISEASE
Neoplastic diseases
Avian neoplastic disease includes a variety o f conditions with one
com m on feature: their tum oral nature. The econom ically m ost
im portant diseases are M areks disease, caused by a herpesvirus,
Leukosis-sarcoma and Reticuloendotheliosis, caused by retroviruses.
32
Marek's disease
M arek s disease (M D ) is a com m o n lym phoproliferative d isease o f
chickens, caused by a cell-associated Herpesvirus, w ith lym phottopic
properties sim ilar to 7-Herpesvims, b u t a m olecu lar structure sim ilar
to a -Herpesvirus, w here it is actually classified.
T hree serotypes o f M D virus have been dem onstrated; only serotype
1 (M DV-1) is path ogen ic, causing tu m ou rs; the oth er two serotypes
(M DV-2 an d H V T-3) are non -on cogen ic an d ate used as vaccines.
Sin ce its discovery, MDV-1 has increasingly enhanced its virulence,
From m ild (m) to very virulent plus (w + ); at the m om en t, virulence
Lymphoid proliferation and enlargement
of tne peripheral nerve (neck) in MD:
a) normal
b) affected
seem s to be fairly stable o r progressin g slowly.
bpizootioiogY
M D is w idespread through out the world. T he prevalence o f infection
is m uch higher than the prevalence o f the disease. C h ick en s are by
far the m ost im portant natural host, with wide differences in genetic
su sc e p tib ility a m o n g th e b reed s. N a tu ra l o u tb re ak s o f M D are
relatively com m on also relatively com m on in Japanese quail. T um ours
have o c c asio n ally been re p o rte d in turkeys, bu t, m ore recently,
severe o u tb reak s have been rep orted in these bird s in Fran ce an d
Israel. M D has also been reported in ph easan ts.
T ra n sm issio n o f in fection occu rs on ly horizontally, by d ire ct or
in direct con tact betw een birds, ap p aren tly by the airb o rn e route.
T he sources of environm ental con tam in ation are the epithelial cells
o f feath er follicles, where the com plete virus replicates.
T h e keratinized d an d er an d feathers, which preserve the virus even
for several m onth s at Z 5 C an d for som e years at 4 C , are the m ain
virus shedder. U n d e r field con dition s, particularly it houses are not
cleaned and disinfected carefully, youn g chicks are usually exposed
to in fection very rapidly, w hich sp read s quickly from bird to bird.
T h e v iru s p e r sists in in fe c te d a n im a ls fo r a lo n g tim e, o fte n
indefinitely, also in the form o f latent in fection in T-lym phocytes.
Factors affecting the developm en t o f the disease are; virus virulence,
Great sciatic nerve enlargement, which is

normally monolateral, in MD
breed genetic susceptibility, environ m ental hygiene, stress, im m une

state, en v iron m en tal tem peratu re (tropics), in teraction with oth er
im m unodepressive viruses, toxins, etc. However, to control M D the
h y gien ic c o n d it io n s o f th e p o u ltry h o u se are o f p a r a m o u n t
33
im p ortan ce, w ith carefu l rem oval of litter an d dust, w ash in g an d

d isinfection, all in-all o u t breeding system s, particularly for the first
few weeks o f life, until the chicks are well protected by the vaccine.
M D is som etim es characterized by progressive paresis and, at a later

stage, by spastic paralysis, asso ciated with lym phoid in filtratio n o f
p erip h eral nerves, m ainly v isib le in the sciatic an d vagu s nerves,
m ostly o n on e sid e . H ow ever, m ore fre q u e n tly th e b ird s sh ow
Spleen and liver tumoral infiltration due to
MD infection
d e p re ssio n , w eigh t lo ss, p alo u r, an o re x ia an d , at n ecropsy, th e

p resen ce o f m ore or less w idespread n e o p lastic lesion s in vario u s
organs, m ainly in the liver, spleen , proven trie ulus, kidneys, go n ad s
and skin.
A tran sien t paralysis syndrom e has been reported in the field, b u t
with lower freq u en cies sin ce v accin atio n yet b ecam e w idesp read .
H ow ever the p ath o g e n e sis o f th is sy n d ro m e has n o t yet becam e
clear. Very virulent M D V strains can induce su d d en death in a few
days in youn g chicks w ith or w ith ou t m acroscop ic tu m oral lesion s
(mostly in chicks experimentally inoculated when they are one-day-old).
M orbidity an d m ortality are rath er variable. T h e usual occurrence
o f d isease, u n d e r field co n d itio n s, is w hen the b ird s are betw een
10 and 30 weeks old; the percentage o f birds sim ultaneously affected
d oes n o t exceed 2-3% . T h e in cid en ce, befo re the use o f vaccines,
was very variable: losses in flocks o f breeders or layers were estim ated
to range from a few birds to 4 0% , occasionally as high as 7 0 % , also
d ep e n d in g on the breeds. In broilers, in ad d itio n to relatively low
m ortality, c o n d e m n a tio n at the slau g h te r h o u se co u ld be fro m
0.5 to 10% , as a result o f c u tan e o u s an d visceral tu m o ral lesion s.
Presently, in m ost countries or areas, nearly all breeders and layers
are vaccin ated ag ain st M D ; for this reason , losses have d ro p p ed to
less than 2 to 4% , often practically zero. Broilers, which are vaccinated
in m any cou n tries, may experien ce very low sp ecific m ortality and
co n d e m n atio n (0.1-0.5% ).
Breeder with tumoral lesions due to MD in

a) muscles and b) spleen
Diagnosis
D iagn o sis o f M D has lon g been p articu larly b ase d on m acro- an d
m icroscopic lesions in tissues. A fter the discovery that feather follicle
ep ith eliu m (FFE ) is the m ain site o f active M D V rep licatio n , the
agar-gel d iffu sio n (A G D ) test was also applied, u sin g a h om ogen ate
o f F F E o r feath er tip s (F T ) as an tig e n , a g a in st a h y p e rim m u n e
an tiserum as reference antibody. T h e test also m akes it p ossib le to
differen tiate betw een p ath ogen ic an d non p ath o gen ic (or vaccinal)
34
stra in s o r seroty p es o f th e viru s, b e c au se F F E , in fected w ith the

latter types o f virus, rarely in du ces any positive reaction; th is is due,
in all lik elih o o d , to a low viru s re p lic atio n a n d c o n c e n tratio n in
F F E . T h e reac tio n u su ally p e rsists u n til th e d e a th o f th e bird .
It h as also been d em o n strated th at there is a sign ifican t relationship
between the percentage o f positivity to th e A G D test, m ainly w hen
perform ed at 12-16 weeks o f age, an d the estim ated cum ulative M D
m ortality d u rin g th e p ro d u c tio n cycle. T h ere fo re, th e u se o f the
A G D test w ith F T d eserv es p articu lar atte n tio n fo r a sim p le and
Breeder chicken with cutaneous tumoral
lesions in MD
accurate diagnosis, feasible in all laboratories, b u t also fo r its reliable

p ro gn o stic valu e o n fu tu re M D in cid e n ce in a flo ck o f long-lived
bird s. S o it c o u ld be o f valu e, a t least indicatively, if n o t legally, in
the sale o t p ullets, a fre q u e n t practice in several cou n tries.
R ecen tly a P C R test has b een u sed to d iffe re n tiate th e serotypes.
Control
V a c c in a tio n rep re se n ts, now an d fo r th e fo re se eab le fu tu re , the
m ain strategy fo r the prevention an d con trol o f M D . S e le ctio n for
gen etic resistan ce an d bio secu rity m easu res w ere u sed a s th e so le
tool available before, the discovery an d ap p lic atio n o f the vaccines,
and subsequently as ad ju n cts to vaccination. O bviously, the efficacy
o t the vaccine is higher in genetically m ore resistant breeds; therefore,
a reco n sid eratio n o f gen etic selection is in progress, particu larly in
very sen sitive lin es o f chickens.
V accin atio n is p e rfo rm e d w hen ch icks are one-day-old, m ostly at
the hatchery, b ecau se early im m u n ity is essential. R evaccination at
o n e day o r at 7 d ay s is so m e tim e s rec o m m e n d e d , p articu larly to
v accin ate b ird s w hich m ig h t have b een sk ip p e d (their percen tage
c o u ld b e as h igh as 5 % o r even m ore) o r to reinforce im m unity.
Vaccines are adm inistered by m eans o f intram uscular o r subcutaneous
in jection . In b ig h atch eries, particu larly in bro ilers, th e vaccin e is
o fte n a d m in iste re d in-ovo o n th e 18th d ay o f in c u b a tio n , th u s
p o s tp o n in g e x p o s u r e to w ild v iru se s fo r a t le a st 3 d ay s a fte r
Lymphomatous lesions in the proventriculus
in MD
vaccination. A t least 7 days are required to establish a solid im m unity

afte r vaccination.
A s regard s the ch o ice o f vaccin es, H V T a lo n e p ro v id es ad e q u ate
p ro tec tio n to b ro ile rs u n d e r n o rm a l e p id e m io lo g ic al c o n d itio n s,
b u t a bivalent vaccine is o fte n required, c o m b in in g serotype 3 with
serotype 1 o r 2, particularly again st w M D V . In b reed ers an d layers,
the use o f the original C V I-9 8 8 o r R isp en s strain o f M D V -1, alon e
o r in a sso c ia tio n w ith H V T , is re c o m m en d e d nearly everyw here.
H V T v a c c in e is av ailab le a s cell-asso ciated (frozen ) o r cell-free
35
NEOPLASTIC DISEASES - MAREK'S DISEASE
(freeze-dried) types; M D V seroty pes 1 a n d 2 are av ailab le only in

c ell-asso c ia ted fo rm , p re se rv e d in liq u id n itro g e n . In th is case,
p ro p er h an d lin g o f th e vaccine d u rin g thaw ing, recon stitu tion and
in je ctio n is very im p o rta n t, in o rd e r to p reserv e cell integrity,
viability, a n d , consequently, the v accin al titre.
M D o u tb reak s in vaccin ated flo ck s m ay o c c u r fo r variou s reasons:
im proper storage, h an d lin g an d in jection o f vaccine, a lso a sh orter
in terval betw een v a cc in atio n a n d e x p o su re to w ild viru ses, weak
v accin es such as H V T again st w M D V , higher gen etic susceptibility
AGD test with leather tips: precipitation line
versus positive serum
o f som e breeds to the disease, im m unosuppressive stress, particularly

d u r i n g t h e o n s e t o f e g g la y in g , i n t e r a c t i o n w ith o t h e r
im m u n o su p p re ssiv e v iru ses (1BDV, C A V , R E V , etc.) a n d , finally,
certain u n kn o w n vagaries.
AGD test with leather tips: precipitation line

:56
NEOPLASTIC DISEASES - LEUKOSIS/SARCOMA GROUP
Leukosis/Sarcoma group
T h e le u k o sis/sa r c o m a ( L / S ) d ise a se g ro u p in c lu d e s a variety o f
b e n ig n a n d m a lig n a n t n e o p la sm s o f p o u ltry c a u se d by c e rtain
m e m b e rs o f th e fam ily Retroviridae, g e n u s Oncovirus. T h e m o st
com m o n o f them arc lym phoid and, m ore recently, myeloid leukosis.
T h e y p o sse s th e enzym e reverse tra n sc rip ta se , w h ich d ire c ts the
syn th esis o f th e p roviral D N A fo rm fro m viral R N A in th e cclls,
w here it in tegrates in to th e g en om e.
O n ly certain d isease s o f th is g ro u p have any sig n ifica n t eco n o m ic
im p o rtan ce, particu larly th o se d u e to av ian leu k o sis v iru s (A LV ),
w ith so m e m ortality, less th an 2 % , occasion ally over 2 0 % , o r subclinical in fectio n s with a depressive effect o n v ario u s p e rform an ce
factors, in clu d in g egg p ro d u c tio n an d quality. T h e ir p u blic health
sign ifican ce h as n o t been clearly d ete rm in e d , alth o u g h evid en ces
su g g e sts th at A LV ' a p p a re n tly d o c s n o t c o n stitu te a d a n g e r fo r
h u m an s.
Epizootiology
Subgroups A an d J o f A L V a re th e m o st c o m m o n v iru se s c a u sin g
ly m p h o id a n d m y elo id le u k o sis (L L o r M L ) resp ectiv ely; o th e r
s u b g r o u p s (B , C , D ) h av e b een iso la te d , m o stly sp o r a d ic a lly .
A n tib o d ies to so m e o f them have also been ob served in wild birds.
E xo gen ou s A L V s are transm itted vertically by the egg to the progeny
o r h o rizo n tally by d ire c t o r in d ire c t c o n ta c t fro m b ird to b ird .
A lth o u g h vertical tra n sm issio n is n o t fre q u e n t, th is m e th o d o f
tra n sm issio n is very im p o rta n t, b e c o m in g a m e a n s to m ain tain
in fection , gen eration after gen eratio n . A L V is p resen t in albu m en
an d tran sm issio n to the em bryo is d u e ro the e lim in atio n o f virus
by the glan d s o f the oviduct. F our differen t classes o f A L V infection
a re re c o g n iz e d , w ith o r w ith o u t v ire m ia a n d w ith o r w ith o u t
sim u lta n e o u s p rese n c e o f viru s a n d a n tib o d ie s. T h e in cu b atio n
p erio d o f the d ise a se d e p e n d s o n th e strain an d q u an tity o f virus,
the route o f infection, the age at exposure and the genetics o f the host.

T h e se in clu d e in ap p e te n c e , w eak n ess, d ia rrh e a , em ac iatio n a n d
th ic k e n in g o f th e lo n g b o n e s o f th e lim b s (o ste o p e tro sis), etc.
37
NEOPLASTIC DISEASES - LEUKOSIS/SARCOMA GROUP
L esio n s d e p e n d on the cells involved an d in clude n eo p lasm s in th e

liver, sp le e n , b u r s a o f F a b r ic iu s, k id n e y , lu n g , m esen tery , etc.
(L L , e r y t h r o b l a s t o s i s , m y e lo c y t o m a to s is , n e p h r o b la s t o m a ,
fibrosarcom a o r osteopetrosis, etc.); tum ours in the bursa o f Fabricius
are p ath o gn o m o n ic o f A LV .
Diagnosis
V iru s iso latio n an d d e m o n stra tio n o f the an tig en o r an tib o d y are
sig n s o f in fe c tio n , b u t n o r o f d ise a se . M any la b o rato ry tests are
Characteristic tumoral lesions in inner sternum
and costochondraljunction o f ribs in J Leucosis
av ailab le fo r the d etectio n o f viru s an tigen o r an tib o d ie s. S am p le s

to c o n tro l in clu d e a lb u m e n , b lo o d , se ru m , m e co rn iu m , clo acal
sw ab s, em b ry os, tu m o u rs etc; th e m o st im p o r ta n t te sts are th e
C O F A L , E L IS A , R IF a n d P C R tests, etc.
Control
C o n tr o l is o b ta in e d by e r a d ic a tio n , p a rtic u la rly fro m p rim ary
breeding stocks. C o m p an ie s p ro d u cin g b ird s o f this type have m ade
sig n ifica n t p ro g resses, in red u cin g o r erad ica tin g A L V su b g ro u p s
A , B and J; in com m ercial stocks, the application o f these program m es
is m ore com plicated an d to o expensive. Eradication is based m ainly
o n the test fo r th e an tigen in a lb u m e n an d fo r th e a n tib o d ie s in
serum .
S e le c tio n fo r g e n e tic resistan ce is in p ro g re ss, b u t is very d ifficu lt
to apply.
N o com m ercial vaccin es are av ailab le fo r in fection w ith A L V ; the
id ea h as b e e n e x tre m ely attractiv e , even i f th e d a m a g e d u e to
in fection is norm ally q u ite low an d occasion al.
NEOPLASTIC DISEASES - RETICULOENDOTHELIOSIS
Reticuloendotheliosis
R e tic u lo e n d o th e lio sis (R E ) is a g ro u p o f p ath o lo g ical sy n d ro m e s
caused by a m em ber o f the fam ily Retroviridae in several avian species:
th ese in clude ranting disease, chronic neoplasia and acute reticulum cell
neoplasia. T h e re is little o r n o evid en ce fo r h u m a n in volvem ent.
Epizootiology
R E V in fection is c o m m o n , b u t n o t u b iq u ito u s. In c o n trast, the
in cidence o f R E V -associated clinical disease in com m ercial poultry
varies from sp o ra d ic to negligible. H orizon tal tran sm issio n o ccu rs
by c o n ta c t, d ire c t o r in d ire c t (also th ro u g h in sects); n ovel viral
reserv oirs, su c h a s fow l p o x v iru s a n d M D V , w h ich m ay c o n ta in
in fe c tio u s c lo n e s o f R E V . c o u ld a ls o b e c o n s id e r e d . V e rtic a l
tran sm issio n from an im als w ith p ersisten t v irem ia m ay occu r, b u t
usually at low frequency.
Clinical sign and lesions

Birds m ay be stunted an d pale and so m e o f them may have abnorm al
fe a th e rin g (n a k a n u k e ); m o rtality is rare. V a r io u s p a th o lo g ic a l
situ atio n s m ay be observed: 1) mnting disease: atroph y o f the thym us
a n d B F , ru n tin g, p ro v e n tricu litis, etc. 2) bursal lymphoma: typical
bursal tu m our: 3) n o n bursal lym phom a in variou s organs; 4) turkey
lymphoma: in filtr a tio n o f th e liver, s p le e n a n d o t h e r o rg a n s;
5) multiple syndromes.
Diagnosis
D iag n o sis is especially based o n the d em o n stratio n o f R E V by P C R
o r an tib o d ie s by E L IS A . T h e p ath o logy o f R E V -in d u ced tu m o u rs
can b e c o n fu se d with M a rek 's d ise a se an d ly m p h oid leukosis.
Control
N o p ro c ed u res have, u n til now , been ap p lied in farm s, p artly d u e
to th e sp o ra d ic n atu re o f the d isease.
39
IN F E C T IO U S C H IC K E N S A N A E M IA
Chicken Infectious Anaemia

C h icken infectious anaem ia (C IA ) is caused by a Gjiownts, a m em ber
o f the fam ily Circoviridae; ap p aren tly n o an tigen ic d ifferen ces have
b een recognized betw een th e v a rio u s iso la te s, alth o u g h o n e h a s
recently b een hypothesized in the U S A . T h e viru s, isolated fo r the
first tim e in Ja p a n in 1 9 7 9 , w as so o n rep orted w orldw ide.
Epizootiology
C h ic k e n s are the on ly know n h o st o f the viru s. A lth o u g h b ird s o f
all ages are susceptible to infection, the disease on ly occurs in chicks
u p to 2-3 weeks o f age. W h en b reed ers becom e infected d u rin g rhe
Appearance o f normal 7 day-old chick (right)
and affected by CIA virus (left)
laying p erio d , they tran sm it the viru s vertically fo r 2 to 4 w eeks o r

lo n g e r to th e ir p ro g en y , w h ich sh o w sev ere a n a e m ia a n d h igh
m ortality (10-20% o r m ore). H orizon tal tran sm issio n o f in fection
o c c u rs a t an y age, b u t b ird s fall ill on ly u p to a few w eeks o f age.

C lin ical signs generally d evelop 10-14 days after infection. T h e only
specific sig n s o f d isease are d ep re ssio n an d an a e m ia, w ith p eak s at
14-21 days; h aem ato crir v a lu e s ran ge fro m 8 to 2 8 % (in healthy
b ird s it is n orm ally over 3 0 % ).
G ro ss lesion s are characterized by atrophy o f rhe thym us, som etim es
with c o m p lete d estru ction , d isco lo u re d aplastic b o n e m arrow , well
evaluable in th e fem u r, re d u ce d bu rsa, sw ollen an d m ottled liver.
H e m o rrh ag e s a n d n e c ro sis a re o fte n p resen t, p articu larly in the
wings, m ost likely d u e to secondary bacterial infections (septicem ia,
blue-wing).
C IA virus, inducing dam age to im m une tissues, is obviously extremely
im m unosuppressive, also interacting with o th e r im m unosuppressive
viruses, su c h as M D V , IB D V , R E V .
Diagnosis
Presum ptive d iagn osis o f C I A is based o n epidem iological d ata and
Thimus in normal chick, 3 week-old
lesio n s (th ym u s atro p h y an d an ae m ia). C o n firm a tio n is given by

isolation o f the virus on M S B ! cell-culture lines fo r abou t 10 passages,
until cell d eath is ob served , o r by in o cu latio n in to su scep tib le oneday-old S P F ch icks, to rep rod u ce th e d isease. T h e E L IS A a n d V N
tests are th ose m o st c o m m o n ly u se d fo r an tib o d y d etectio n .
40
Control
T h e c o n tro l o f C I A is b a se d o n th e p re v e n tio n o f vertical virus
tran sm issio n by im m u n izatio n o f b ree d e r flocks. C o m m e rc ial live
vaccines, m ore o r less atten u ated , arc available in several countries.
V a c c in e sh o u ld b e a d m in is te r e d via rh e d r in k in g w ater o r by
in jection at ab o u t 12-15 w eeks o f age, bur never later than 3-4 weeks
before the first collection o f h atch in g eggs. A sin gle ad m in istration
is su fficien t to cover th e en tire laying p erio d.
Atrophy o f the thymus in 3 week-old chick

affected by CIA
41
R E O V IR U S IN F E C T IO N S
Reovirus Infections
A vian reoviru s, a m em b er o f fam ily Rcoviriclae, is o fte n u b iq u ito u s
in com m ercial pou ltry; it is co m m o n ly fo u n d in th e digestive and
respirator^' tract. T h e v a rio u s strain s can b e d ifferen tiated by their
relative p ath o gen icity an d an tig e n ic ch aracteristics, bein g isolated
from b ird s affected b y asso rted disease con d itio n s. T h e d iseases are
g ro u p ed in to two m ain categories:
a) arth ritis/te n o sy n o v itis;
b) stu n tin g o r m alab so rp tio n syn d rom e, w hich in cludes p ale bird s,
a b n o rm a l fe ath e rin g sy n d rom e (h elicop ter d isease), etc.
Tenosynovitis in the tibio-tarsal joint

after infection
Epizootiology
Viral arthritis o r tenosynovitis is a n econ o m ically im p o rta n t d isease,
m ostly in ch ickens fo r m eat p rod u ction (breeders an d broilers), but
it h as also b een ob served in layers an d in turkeys. T h e d ise a se is
cau sed by d ifferen t seroty pes an d path o types o f reovirus; however,
th e best know n and m o st widely stu d ied isolate h as been the strain
S I 133 o f v a n d e r H eyde. H orizontal tran sm issio n o f reo v iru s h as
been extensively d o cu m en ted , b u t a vertical tran sm issio n h as been
clearly d em o n strate d .
Malabsorption syndrome is con sid ered a m ulti factorial d isease, w here
d ifferen t serotypes o f reovirus an d other viru ses have been isolated.
T h is syndrom e is hardly reproduced, in laboratory condition s, w hen
reovirus on ly is in ocu lated .

In arthritis - tenosynovitis, th e in cu b atio n p e rio d v aries, d e p e n d in g
o n th e viru s p ath o ty p e , h o st ag e , ro u te o f e x p o su re a n d type o f
breed; so m e tim e s in fection is u n ap p are n t a n d d e m o n strab le only
by serology o r v iru s iso la tio n . T h e affected b ird s sh ow lam en ess
with swelling, oe d e m a and in flam m ation o f the tibio-tarsal-meta tarsal
Lesion in the synovial sheath
te n d o n sh e a th s, h ig h lig h te d by p a lp a tio n ju s t ab ove th e hock;

r u p tu r e o f th e g a s tr o c n e m iu s te n d o n is s o m e tim e s o b se rv e d .
M o rb id ity a n d m ortality are very v ariab le ; th e latter is gen erally
2-3% , so m etim e s even reach in g 10% , particularly in m ales; loss o f
grow th an d infertility are also evident.
M alabsorption syndrome is c h aracte rized by re d u c e d a n d u n even
gro w th rate s a n d o th e r d iso rd e rs, m ain ly in d ic a tin g th e varyin g
cffects o n g astro in testin al tract fu n ction . M orbidity, m ortality and
delayed grow th vary greatly.
42
Diagnosis
A p re su m p tiv e d ia g n o sis o f d iffe re n t sy n d ro m e s m ay b e m ad e
ac c o rd in g to sign s an d lesion s. D e m o n stratio n o f th e viral antigen
in te n d o n sh eath s by P C R , im m u n o flu o re sc e n c e , viru s iso latio n
in em b ry o n ated eggs o r tissu e c u ltu re s a n d in o c u la tio n in to the
foot-pad of su sc e p tib le I -day-old ch icks (sw elling) provid e furth er
ev id e n c e o f a rth ro sy n o v itis d u e to re o v iru s. A n tib o d ie s c a n be
d etected by the A G P a n d E L IS A rests.
Control
Tenosynovitis in tibio-tarsal tendon sheaths
in heavy breeder, tendon rupture
T h e c o n tr o l o f v iral a rth ritis, b e s id e s th e a p p lic a tio n o f g o o d

b r e e d in g te c h n iq u e s a n d h y g ie n ic m e a s u r e s , is o b t a in e d by
vaccin ation o f b reed ers w ith live atten u ated viru s (strain S 1133) at:
a b o u t 7 days o f age, follow ed by a b o o ste r w ith live o r in activated
vaccine in oily e m u lsio n , w hen b ird s are a b o u t 18 w eeks old.
T h e ch ick s b atch e d fro m th ese b reed ers are protected by m aternal
an tib o d ie s fo r 2-3 w eeks, a n d sh o u ld on ly b e vaccinated in case o f
high viral pressure.
A tte m p ts to c o n tro l m a la b so rp tio n sy n d ro m e h av e b een m ad e ,
with very v ariab le resu lts, u sin g d ifferen t sero ty p es o f v iru s, o fte n
in polyvalent in activated vaccin es in oily em u lsio n .
Swelling (tenosynovitis) oftarsal-metatarsal

tendon sheaths in pullets:
a) normal
b) with lesions
A D E N O V IR U S IN F E C T IO N S
Adenovirus Infections
A denoviruses, m em bers o f the fam ily Adenoviridae, genus Aviadeiuwirus
are co m m o n in fectiou s agen ts in poultry w orldw ide. M o st o f these
viru ses are p rese n t in h ealth y b ird s w ith n o sig n s o f in fectio n , or
c an beh ave a s o p p o rtu n istic p ath o g e n s, w h en a d d itio n a l factors,
either in fectious o r not, are involved. H ow ever, so m e aden oviru ses
are p rim ary p a th o g e n s, su ch a s th o se c a u sin g turkey hemorrhagic
enteritis, pheasant marble spleen disease, chicken egg-drop syndrome and
quail bronchitis virus.
SUB-GROUP I ADENOVIRUS
T h e role o f these viru ses a s p ath o gen s is n o t well d efin e d , with rhe
e x c e p tio n o f q u a il b ro n c h itis, h y d ro p e rica rd iu m sy n d ro m e an d
in clu sio n b o d y hepatitis.
Inclusion body hepatitis (IB H ) is n o rm ally o b se rv e d in m eat-type
ch icken s a t 3-7 w eeks o f age and is characterized by a su d d e n o n se t
o f m ortality fo r 3-5 days, w h ich m ay reach a th resh o ld o f 10% o r
even h igh er. M an y se ro ty p e s h av e b een a sso c ia te d w ith n atu ral
ou tb reaks o f d isease. T h e m ain lesion s are pale, friable an d sw ollen
liver, w ith p in p o in t h aem orrh agic a n d necrotic sp ots.
Hydropericardium syndrome (H IS), o r A n g ara d isease, w as recognized
a s a d e v a sta tin g d is o r d e r in b ro ile r s in P a k ista n in 1 9 8 7 an d
su b seq u en tly in o th e r co u n tries. It affects bird s 3-4 w eeks-old an d
is ch aracterized by a n a c c u m u latio n o f clear, straw -coloured flu id
in the p ericardial sac, p u lm on ary o ed em a, sw ollen liver an d kidney.
Experim ents with vaccination, apparently successful, with inactivated
liver h om ogen are ob tain ed from infected birds, have been reported
in Pakistan.
Q uail bronchitis (Q B ) is a disease o f econ om ic significance w orldwide,
characterized by rapid o n set, w ith high m orbidity' an d mortality' in
b ird s o f less th an 3 w eeks o f age. S ic k b ird s sh ow ruffled feath ers,
d iffic u ltie s in b re a th in g , n asal-ocu lar d isch arg e. T h e m ain g ro ss
lesion s arc localized in rhe respiratory tract. Prevention is based on
the usual san itatio n p ro ced u res an d biosecurity' m easu res; to date,
n o vaccin es are available.
SUB-GROUP II ADENOVIRUS
This subgroup includes haemorrhagic enteritis and marble spleen disease.
14
Haemmrhagic enteritis (H E ) is a n acute o r su bacu te disease o f turkeys,

w hich co m m o n ly affects 7-IQ-week-old bird s. It is characterized by
a rapid p rogression o f clinical sign s, in cluding d epression , catarrhal
to bloody droppings and death, high m orbidity an d variable mortality
(from less then 1% to 6 0 % , w ith an average o f 10% ). A t necropsy,
the sm all in testin e is co m m o n ly d isten d ed a n d filled w ith b loo d y
o r catarrh al co n ten t, th e sp lee n is en larg ed an d m ottled.
E ith er live an d in activated vaccin es are available.
Live vaccines are generally adm inistered in drin k in g w ater to healthy
turkeys a t 4-6 w eeks o f age.
Quality o f a) normal eggs; b) variously altered
(thin or shell-less) eggs in EDS
A n in activated vaccin e in oily em u lsio n is also in use, particularly

in E u r o p e , w ith tw o in je c tio n s a t 4-6 a n d 8 -1 0 w eeks o f age.
Marble spleen disease (M S D ) is a n acu te d isease o f p h e asan ts, w hich
occurs in birds o f 3-8 m onths of age. It is characterized by depression
a n d p ro g re ssiv e d y sp n e a . M o rtality fro m 2 % to 1 0 % h a s b e e n
reported. G ro ss lesion s co n sist o f enlarged an d m arbled sp leen and
con gested lungs.
Live av iru len t vaccin es, ad m in iste red in w ater, a re e ffica cio u s in
con tro llin g th e d isease.
SUB-GROUP III ADENOVIRUS

Egg drop syndrome (E D S ) virus is the so le m em ber o f the g ro u p : only
o n e sero ty p e is recognized a n d it is h aem ag g lu tin a tin g . S in c e its
initial report in T h e N eth erlands in 1976, rhe E D S virus has becom e
a m ajo r cause o f losses in egg p ro d u c tio n , w orldw ide. T h e viru s is
tran sm itted vertically, th rou gh em bry on ated eggs a n d horizontally
in sid e th e flo c k a n d fr o m flo c k to flo c k , by d r o p p in g s a n d
c o n ta m in ate d egg trays. T h e v iru s o fte n rem ain s late n t until the
b ird s h av e reach ed a b o u t 5 0 % egg p ro d u c tio n ; a t th is stag e the
virus is reactivated, resultin g in a rap id sp re ad o f inlection.
B e c au se o f the ab se n ce o f a n tib o d ie s to the v iru s in ch ick en sera
p rio r to 1 975, a n d its o p tim a l grow th in d u c k cells, it h a s been
suggested th at its origin lies in d u ck s, as w as co n firm ed by isolating
th e viru s fro m n o rm al d u c k s a n d g e e se in th e fie ld . T h is viru s
p ro b a b ly u n d e rw e n t an a d a p ta tio n to c h ic k e n s in E u ro p e , the
d isease n o t being present in N orth A m erica, w here immunologicallyrelated v iru ses have also b een iso lated from m an y b reed s o f duck.
C o n ta m in a tio n o f M a re k s d ise a se vaccin e, p re p are d w ith d u ck
em bryo fib ro b lasts, h as n o t been exclu d ed .
T h e first signs o f disease are a reduction in egg colou r an d resistance,
q u ick ly fo llo w e d by p ro d u c tio n o f th in , soft-sh elled o r shell-less
eggs. T h e fall in egg p ro d u c tio n is very rap id in floor-reared birds,
less in cages, an d lasts 4-10 w eeks, w ith a red u ctio n o f u p to 5 0 %
45
- 4,
A D E N O V IR U S IN F E C T IO N S
o r m ore. N o och er sign s, except fo r a p o ssib le tran sien t d iarrh oea,

o r m ortality, are detected.
E D S c a n b e effectively c o n tr o lle d by in je c tin g p u lle ts w ith an
in activated vaccin e in oily em u lsio n , p e rfo rm e d on ly o n ce, a t the
age o f 14-18 w eeks.
Diagnosis
D ia g n o sis o f A d e n o v iru s in fe c tio n s is b a se d o n sy m p to m s an d
lesions, b u t isolation o f th e virus from excreta or tissue h om ogcnates
Histological lesions with intranuclear inclusions
in epithelial cells o f the oviduct plica in EDS
is o fte n necessary o n ch ic k e n em bry o liver o r k id n e y cell tissuec u ltu res. G ro u p -sp e cific-an tib o d ie s can b e d e te c te d by th e A G D
o r E L IS A tests; in case o f E D S , th e H I te st is also used.
L a yin g curve in va ccin a te d a n d c o n tro l la yers , c h a lle n g e d w ith a

v iru le n t strain o f FDS virus
46
A V IA N E N C E P H A LO M Y E LIT IS
Avian Encephalomyelitis
A vian encephalom yelitis (A E ) is cau sed by a n enterovirus, a m em ber
o f the fam ily Picornaviridae. It usually affects very y ou n g ch icks, less
th an 2 w eeks o ld , an d h e n s in lay.
Epizootiology
A E o c c u rs v irtu ally all over th e w o rld . C h ic k s , tu rk eys, q u a il,
p h e a sa n ts a n d guinea-fow l are kn o w n to b e su sc e p tib le . U n d e r
n a tu ra l c o n d itio n s, A E b e in g a n e n te ric in fe c tio n , th e v iru s is
excreted w ith th e fae c e s fo r 1-2 w eeks o r m ore in y o u n g b ird s.
Twelve day-old chicks vertically infected
byAEV
H o riz o n ta l tr a n s m is s io n o c c u rs by in g e stio n o f feed o r w ater

co n tam in ated by faeces. V ertical tran sm ission is the m ain an d m ost
d am ag in g rou te o f in fection . B reed ers, in fected d u rin g th e laying
p erio d, tran sm it the viru s to th eir progeny fo r a t least 4-5 w eeks o r
m ore.

In y oun g chicks, 1-3 weeks old, the disease is characterized by flaccid
paralysis; w eakn ess m ay also b e observed a t hatching. A progressive
ataxia o ccu rs u n til th e chicks are totally p aralysed , p ro strated and
d ie; h ead a n d neck tre m o r m ay o fte n be present.
T h e usual m orbidity rate is to 6 0 % , m ortality is over 10% an d may
exceed 4 0 % , d e p e n d in g o n th e p e rio d o f lay w h en the in fection
o c c u rs in paren ts.
In breeders, infected du ring the laying period, A E is only characterized
by a variab le d ro p in egg p ro d u c tio n (w ith ou t any ch an ges in shell
quality), red u ced hatchability a n d tran sm issio n o f the v iru s to the
progeny. In laying hens, a variable (20 to 4 0 % ) d rop in egg production
occu rs, w hich rapidly returns to n o rm al p ro d u c tio n in 2-3 w eeks.
T h e re are on ly m ic ro sc o p ic le sio n s to th e cen tral n erv o u s system :
Eighteen day-old embryos, normal or affected
byAEV
non-purulent in flam m atio n with a striking perivascular lymphocytic

in filtration .
Diagnosis
A taxia a n d rapid trem ors o f the head in young chicks and a su d den
d ro p in egg p r o d u c tio n in laying h en s, w ith n o ch an g e s in sh ell
quality, is in dicative o f A E . H isto logical e x am in a tio n o f th e brain
48
an d p an c reas reveals th e typical le sio n s o f d ise a se. R e p ro d u c tio n

o f th e d ise a se in S P F chicks, in ocu lated by th e in rracerebral rou te
a t one-day-old, is a c o n firm a tio n . T o d em o n strate an tib o d ie s, the
E L IS A o r V N tests are th e m e th o d s o f choice.
Control
T h e c o n tro l o f A E is ach ieved by v a c c in a tio n o f b re ed e r flo c k s
(so m e tim e also layers) d u rin g th e grow in g p e rio d , a t 12-15 w eeks
o f age, to en su re th at they b e co m e infected an d develop an tib o d ies
Legs o f embryos affected or not by AEV
before g o in g in to lay, thereby preventing virus sh e d d in g by rhe eggb o rn e route. M atern al an tib o d ies protect the progeny from con tact
with A E virus d u rin g th e critical first two weeks o f life.
T h e m o st com m on ly used com m ercial vaccines are prepared m ainly
with the strain C aln ek 1143, only partially attenuated in em bryonated
eggs; obviously, th e viru s m u st m ain tain the sp re ad in g ability and
e n tero trop ism o f wild viru s, so it m u st n o t becom e to o ad ap te d to
th e em bryo. T h e ad m in istratio n m ay b e in d iv id u a l in to the beak
o r by eye-drop o r by d rin k in g water. T o en su re th at birds have been
well im m u n ized , a sero lo g ical check can b e carried o u r 3-4 w eeks
after v accin ation .
Laying curve in vaccinated a n d control hens, infected with virulent AEV
49
Paratyphoid Infections
T h e sp ecies o f g e n u s Salmonella, (S) o f th e fam ily Enterobacteriaceae
w hich a rc m o st im p o r ta n t in a n im al a n d h u m a n p a th o lo g y are
S. enteritidis an d S. cyphimurium; nevertheless, m any o th e r sp ecies o f
S. can occasion ally he involved in d ise a se . H owever, th e d egree o f
genetic relatedness am o n g Salm o n ellae is so great that it has actually
b een su ggested th at m o st o f th em b e g ro u p ed in a sin g le sp ecies,
S. enterica, with different serotypes (over 2400). T h e various serotypes
are c la ssifie d ac c o rd in g to so m a tic (O ) c a p su la r (K) an d flagellar
(H ) antigens.
In so m e instances, these infections result in relatively asymptomatic,
in te stin a l c arriag e , in o th e r s they p ro d u c e c lin ic a l d ise a se an d
m ortality. Progress in p o u ltry p ro d u c tio n p ractices have b ro u gh t
the spread o f S. infection in breeding farm s, with relative consequences
for th e h ealth n o t on ly o f an im als, b u t also o f h u m an s.
T h erefore, the con trol o f S. in fection in p ou ltry flock s h as becom e
an im p ortan t objective from b o th an e co n o m ic and a p u blic health
p ersp ectiv e. In fa c t, th e to ta l c o s t o f m e d ic a l care a n d lo ss in
productivity resu ltin g from food-borne $. in fection s in h u m an s has
been very substantial. However, control m easures, such as biosecurity,
c le a n in g a n d d isin fe c tin g o f fa c ilitie s, te stin g a n d v a c c in a tio n
practices, can in crease p ro d u c tio n c o sts o f p ou ltry eggs an d m eat.
Salm o n ella is a non-spore-form ing, G ra m negative rod, 1x3 m icrons
in size, u su ally m o tile d u e to p e ritric h o u s tlag ella, m u ta n t nonm o tile ro d s are o c c a s io n a lly e n c o u n te r e d , b e sid e s th e c la ssic
n on -m otile sp e c ie s S. pullorum an d S. gallinarum, w hich in d u c e an
acute o r chronic septicem ic disease only in poultry an d are discussed
in a sep arate chapter. A ll S. are facultative an ae ro b e s an d can grow
w ell u n d e r a e r o b ic c o n d itio n s , a t a n o p tim u m te m p e r a tu re o f
3 7-45G .
S. have been fo u n d in both broiler an d layer flocks, in very variable

p ercen tages. A lth o u g h 2 4 0 0 serotypes o f S. have b e e n id en tified ,
only ab ou t 10% o f these have been isolated from poultry, particularly
S. enteriiidis, S. typliimurium, S. thomson, S. nadar, S. seftemberg,
S. blockley, etc..
S. enteritidis is also the m ost com m o n serotype responsible for h u m an

in fectio n s, w hich have in creased c o n sid e rab ly a n d grad u ally over
the last 3 0 years, worldwide. Paratyphoid infections in very susceptible
youn g chicks an d in o th e r d o m estic species are also asso ciated with
m ortality ran gin g as high as 5 0 % . O ld e r bird s are con sid erably less
su sceptible an d the bacteria colonize their gut. T ran sm issio n occurs
particu larly by th e h orizo n tal ro u te , sp re a d in g by c o n ta c t vectors
(m ice, insects, m ealw orm s, etc.) and con tam in ated feed, o r vertically,
by extern al an d in ternal tran sovarian egg c o n ta m in atio n .
Lesions in septicemic form o f infection:
spleen and liver
S. in fection in poultry is usually associated w ith d isease on ly in very
y o u n g b ird s: m alaise, an o rex ia, d iarrh o ea, em ac iatio n , sep ticem ia
a n d variable m ortality, w ith swollen parenchym as an d focal necrotic
lesion s, particularly in the intestine, liver an d spleen . In ad u lt birds,
S. can cau se an o rexia, d iarrh ea an d a red u ctio n in egg p ro d u ctio n ,
usually with low m ortality, with the sam e lesion s observed in young
birds.
A lth o u g h clin ica l a n d p a th o lo g ic a l fin d in g s m ig h t su g g e st th e

lik e lih o o d o f in fe c tio n , a fin a l d ia g n o s is o f S. d e p e n d s o n th e
iso la tio n a n d id e n tific a tio n o f th e cau sativ e o rg a n ism fro m the
tissu e s o f d ead o r live b ird s, feed , feces, e n v iro n m e n t, litter etc.
V ery im p o r ta n t in te r v e n t io n stra te g ie s h av e b e c o m e p a r t o t
m a n a g e m e n t p ro c e d u re s in h atch eries a n d farm s, w ith accu rate
c lean in g , d isin fe c tio n a n d w ise p la ce m e n t o f o n e day o ld ch icks,
ro d en t a n d in sect c o n tro l an d rigid biosecu rity m easures.
T h e S. statu s of p ou ltry an d o f th eir e n v iron m en t sh o u ld be tested
frequently, im p lem en tin g m o n ito rin g p rogram m es particu larly for
S. enteritidis an d S. typhimurium. C om petitive exclusion, u sin g norm al
in testin al b acterial flo ra o r lactob acilli to in h ib it co lo n izatio n by
S. an d o th e r p ath o g e n s, h as served fo r the d e v e lo p m e n t o f furth er
treatm ents.
V accination with killed o r live bacterial preparations has been found

to red u c e su sce p tib ility to , a n d s h e d d in g o f S., alth o u g h n eith er
type o f vaccin e h a s con sisten tly p rovided a n im p e n e trab le b arrier
ag ain st in fection .
51
Live atten u ated vaccines need to p ersist in the tissu es lon g enough
to in d u ce a protective im m u n e respon se.
K illed v a c c in e s (b a c te rin s), p a rtic u la rly if a d ju v a n te d w ith oily
em u lsio n , have been rep orted to significantly redu ce th e in cidence
o f faecal sh e d d in g a n d iso la tio n o f S. enteritidis a n d S. tyl>hiinurium
from v a rio u s tissu es, particu larly the ovary. T h e se latter vaccin es
m u st b e in ocu lated o n ce, a.s a b o o ste r befo re laying a t 16-18 weeks,
afte r a first tre a tm e n t w ith live vaccin es o r tw ice a t 4-6 an d 10-12
w eeks o f age.
T h e efficacy a n d w isdom o f a n tib io tic m e d ic a tio n s ro p reven t o r

treat S. in fection in p ou ltry are to p ics o f co n sid e rab le debate.
M any antibiotics have been used to control such infection, particularly
in y o u n g c h ic k s: q u in o lo n e s , a m p ic illin , a m o x ic illin , c o listin ,
gentam ycin , etc..
PULLORUM DISEASE AND FOWL TYPHOID
Pullorum Disease and Fowl Typhoid

Pullorum disease (PD ) and Fow l typhoid (FT) are im p ortan t diseases
o f fowl, caused by Salmonella enterica, serovar pullorum an d gallinarum,
h av in g th e sa m e an tig e n ic ch arac te ristics (O a n tig e n s 9 , 12, b u t
w ith d ifferen t percentages o f the 12,, 12?, 12?, antigens, n o flagella),
b u t c ertain d iffere n t b ioch em ical p ro p e rtie s. T h ey are sep ticem ic
d ise a se s, prim arily affe c tin g ch ick en s a n d turkeys, b u t o th e r fowl
are also susceptible.
P D a n d F T a re fo u n d w orld w id e, b u t have b een reaso n ab ly well
con trolled in m any co u n trie s, startin g fro m th e m id-20h C en tu ry ,
by sero lo g ical te sts, rep eated a n u m b e r o f tim es if n ecessary an d
w ith rem oval of reactin g b ird s fro m th e flock.
C h icken s are the natural hosts, but oth er species can also b e affected.
Sign ifican t d ifferen ces in susceptibility to PD and F T a m o n g breeds
have b een re p o rte d ; legh orn b reed s a p p e a r to be m ore resistan t.
T h e disease can b e transm itted in two ways: horizontally, by infected
b ird s a n d faeces (re acto rs o r carriers) to o th e r b ird s o f th e sam e
flock o r vertically, th ro u gh egg tran sm issio n (ovules, ovidu ct, shell
p en etratio n , hatching) in varying percentages.
! Clinical signs and lesions

P D a n d F T are p rim arily d ise a se s o f chicks an d p o u lts, b u t F T is
a m ore sig n ifica n t d ise a se in grow in g, a n d p articu larly in lay >ng,
birds.
T h e y o u n g b ird s sh o w w eak n ess, p o o r g row th , a ch alk y w h ite
m aterial a t the ven t, m o rtality from h atch in g until th e th ird week
o f age. S u rv iv o rs m ay b e greatly retarded in th e ir grow th a n d can
show b lin d n e ss, sw elling o f th e jo in ts a n d o th e r lesion s.
In grow in g a n d m atu re b ird s, a c u te o u tb re ak s o f F T m ay occu r,
with m alaise, pale and shrunken com bs, a d rop in feed con su m p tion
a n d egg p ro d u ctio n .
M o rbidity an d m ortality in P D and F T are very variable, d epe n d in g
o n th e m eth o d o f tran sm issio n , th e strain o f S., th e breed o f bird
an d flo ck m an ag e m en t; th ey ran ge from 5 to 5 0 % o r m ore.
M ites, have been sh ow n to be a n im p o rta n t vector o f S. M orbidity
is ob viously high er th an m ortality. A s regards th e lesion s, in chicks
and in acute cases, enlarged an d congested liver, spleen an d kidneys,
54
to g e th e r w ith n e c ro tic foci in th e liver m ay b e o b se rv e d ; p o o r

reab so rp tio n o f the yolk-sac w ith a cream y an d case o u s consistency
o f th e yolk, w h ite n o d u le s m th e m y o c a rd iu m , a r th r itis etc.;
in a d u lt b ird s, le sio n s d u e to P D m ay b e m in im al o r ovaritis may
be present, with m issh apen cystic ovules an d oviductitis, b ecom in g
serological reactors; the lesion s d u e to F T frequently show a n acute
c o u rse w ith en larged liver, sp le e n a n d k idn eys, p e rito n itis, a n d
pericard itis, w ith high m ortality, so m e tim e over 5 0 % .
Visceral lesions in S. aallinarum infection:

enlarged spleen ana liver with necrotic foci
Diagnosis
A tentative d iagn osis is based o n flock history, clinical sign s, lesions,
m ortality an d scrological findings, b u t a definitive d iagn osis is based
o n iso la tio n a n d id e n tifica tio n o f th e cau sative a g e n t in su itab le
m ed ia. T h e sero lo g ical tests in clu d e slid e an d ru be ag g lu tin atio n
and E L ISA . Differential d iagn osis from other S. is based on isolation
o f the m icroorganism o r indirectly o n dem onstration o f the antibody.
C o n tr o l
C o n tro l o f P D is b a sed o n sero lo g ical te sts a t th e sta rt o f laying
w ith e lim in a tio n o f carriers, so m e tim e s rep eatin g the tests u n til
negativity is o b tain ed . N on-specific reactions can occasionally cause
in terp retation p ro b lem s: o th e r S. sp ecies, particu larly $. enreritidis,
o r o th er bacterial in fection s.
T ill now vaccin atio n w ith live (u sin g a rough strain ) o r inactivated
vaccin e gives u n even results.
T h e m o st effective th e ra p e u tic d ru g s are am oxicillin , q u in o lo n e s,
tetracycline, etc..
55
C O L IB A C IL L O S I
Colibacillosis
C olibacillosis is a localized o r system ic infection caused by Escherichia
coli (E. coli), a n o rm al com m en sal bacteriu m o f the lower intestinal
tract o f all an im al species. It can b e a p rim ary or, m ore frequently,
a se c o n d a ry p a th o g e n , w ith v a riab le v iru le n c e , r e sp o n sib le for
sig n ific a n t e c o n o m ic lo sse s, p articu larly in b ro ilers, b u t a lso in
lay ers a n d b re e d e rs. M o st s tr a in s , iso la te d fro m p o u ltry , are
p a th o g e n ic only fo r b ird s, b u t a lso rep resen t a low risk o f d isease
fo r p eop le a n d oth er an im als.
E. coli is a g en u s o f th e fam ily Enterobacteriaceae, G ram -negative,
non-spore-torm ing, m ostly m otile a n d lactose ferm en tin g, grow ing
on o rd in ary n u trien t m edia, facultative an aerobes.
T h e v a rio u s serotypes o f E. coli are classified acco rd in g t o so m atic
(O ), c a p su la r (K ), flag e lla r (H ) a n d fim b rial (F) a n tig e n s; today
a b o u t 170 O , 75 K, 55 H an d 18 F an tig en s have been recognized.
E n d o to x in s, e so to x in s a n d v iru le n ce facto rs a r e a lso p ro d u c e d .
Epi/.cioliology
. coli is a c o m m o n in h a b ita n t o f th e in te stin e o f p o u ltr y at
concentrations u p to 106 m icro o rg an ism s/g o r m ore in faeces; ab o u t
15% o f intestinal eoliform s belong to potentially pathogenic serotypes,
w hich can b e localized o r sp read , u n d er certain con d ition s, to other
tissues an d organ s, particularly to the respiratory an d reproductive
tracts. D u st in poultry houses may con tain 10M 06 m icroorganism s/g;
feed is often con tam in ated , b u t h ot pelleting can destroy eoliform s.
T ran sm issio n o f E. coli o c c u rs m ostly by the o ral a n d resp iratory
ro u te s, c o lo n iz in g p a rtic u la rly th e re sp ira to ry a n d rep ro d u ctiv e
tracts.
S y m p to m s a n d le sio n s
S ev eral lo caliz e d o r sy stem ic fo rm s o f c o lib a c illo s is have been
d escrib ed in poultry, o fte n b ased o n the tissu es affected . T h e m ost
c o m m o n localized in fectio n s are o m p h alitis, salp in go -p erito n itis,
e n te ritis, c e llu lite s; th e m o st c o m m o n sy ste m ic in fe c tio n s are
c o lisep ticem ia (airsaccu litis, m e n in g o e n c e p h a litis, osteom y elitis,
ten o syn ov itis, p e ric a rd itis, etc.) an d c o lig ra n u lo m a . A fte r E. coli
con tacts the h o st tissu es, an acu te in flam m atory resp on se develops
56
with ac c u m u latio n of sero u s, catarrh al, fib rin o u s, p u ru le n t and

c a se o u s e x u d a te s. C o h s e p tic e in ia a ffe c ts m ostly y o u n g b ird s, I
prin cipally b ro ilers, bu t also ad u lt birds, p articu larly at the on set ,
o f egg p ro d u ctio n , due to the stress.
D iagn osis is based on typical, lesions, b u t is con firm ed by isolation

an d id e n tific a tio n o f the c a u sa tiv e a g e n t in se le ctiv e m e d ia
(M acConkey, EM B agar, etc.). Antigenic identification, determ ination
Lesions due to E. coli: liver and spleen
enlargement and necrotic and fibrinous

exudate on liver and pericardium
o f viru len ce factors or fin gerp rin tin g the strain m ight be helpful,
particularly in e p id em iological investigations.
T h e m ost im p ortan t sou rce for the tran sm ission o f infection is the
faecal co n ta m in atio n of h atch ing eggs; for this reason, hygiene o f
laying, in cu b atio n and h atching o f eggs is a fu n d am e n tal tool for
control of colibacillosis. T h e hygienic condition o f houses (ventilation,
litter, water, teed etc.) is also very im portan t.
E. coli is sen sitiv e to m any d ru gs su ch as am ox icillin , am p icillin ,
q u in o lo n e s o f v a r io u s g e n e r a tio n s , te tra c y c lin e s, n e o m y c in ,
sp ectin om ycin , etc. Frequently, iso lates o f E. coli show an tib io tic
re sistan c e , esp ecially if th ese have been w idely or badly u sed .
A n tib iogram s are very im p ortan t in the choice o f the specific drug.
A s regard s im m u n iz atio n , effective in activ ated v accin es ag a in st
v ariou s serotypes have been p rod u ced; a m on ov alen t vaccine gives
the b est results. R coli vaccines are infrequently used, also becau se
of their v ariability' and the need for at least two injections to ob tain
g o o d im m unity.
57
IN F E C T IO U S C O R Y ZA
Infectious Coryza
In fectiou s coryza (1C) is an acute resp iratory d isease o f chickens,
caused by a G ram -negative bacteriu m , Haemophilus paragallinarum.
T h ree serotypes (A, B an d C ) have been re p o rte d ; serotype B is
con fin ed only to certain geograph ical areas.
Epizootiology
T he disease occurs worldwide, in birds o f all ages, but m ore frequently
in adult birds, particularly in m ultiaged farms. U n d er such conditions,
ch ron ic carriers are o fte n the sou rce o f in fection . T ran sm issio n
within an infected flock m ainly occurs by direct contact; from flock
Inflammation of the head: rhinitis, sinusitis,
conjunctivitis and facial oedema in 1C
to flock or from farm to farm by contam inated feed, water, equipm ent
an d p erso n n e l, so m e tim e s by the airb o rn e route. It o fte n occu rs
w hen old h ens are lo a d e d u sin g tra n sp o rt cages an d tru cks n o t
su itably d isin fected , or w h en rep lacem en t p u llets are in trod u ced
to a m ultiaged farm.

H. paragallinamm may affect birds w ith out any sym ptom s; the latter
ap p ear w hen stress factors occur (start o f laying, su d d en change in
environm ental condition s). T h e m ost p rom in en t features o f disease
are: in flam m atio n o f the u p p e r respiratory tract, a greyish m u co id
nasal discharge, con ju n ctivitis, sin u sitis, facial oed em a, a decrease
in feed and water consum ption and a drop in egg production (20-70%).
M orbidity is very high, m ortality is variable, b u t can reach, in som e
flocks, a percentage as high as 20 to 3 0% .
A cu te c atarrh al-fib rin o u s rh in itis an d sin u sitis are the sp e c ific
conjunctivitis and facial oedema in 1C
lesion s, frequently su b c u tan e o u s facial and wattle o ed em a occurs.
Diagnosis
D iagn osis o f IC is based on the rapid sp read o f the disease and on
signs, b u t it is con firm ed by iso latio n o f the bacteriu m . A lth o u g h
H. paragallinarum is considered to be a fastidiou s organ ism to grow,
it is n o t d iffic u lt to iso la te , stre a k in g e x u d a te s, m ain ly fro m
in frao rb ita l sin u ses, on b lo o d agar p lates, w hich are th en c ro ss
streaked with non-haem olytic Staphylococcus and in cubated at 37C :
very sm all tran slu c e n t an d h aem oly tic c o lo n ie s d evelop , m ostly
close to those o f Staphylococcus.
58
Control
T reatm en t o f d isease is b ase d on the ad m in istratio n o f an tibiotics
(am oxicillin, m acrolid es, q u in o lo n e s, etc.) in d rin k in g water.
Sin ce som e carriers o f in fection may rem ain am o n g treated birds,
in o rd e r to e lim in a te the a g e n t fro m a farm , flo c k s m a s t be
depopulated and prem ises and equ ipm en t disinfected or vaccination
o f future rep lacem en ts ad op ted .
G o o d protection is provided by a bacterin, prepared with inactivated
H. p aragallin aru m , c o n ta in in g 2 or 3 seroty p es, a d so rb e d o n to
alum inium hydroxide or em ulsified with m ineral oil. Two injections

conjunctivitis and facia! oedema in 1C
in pullets, at least 4-5 weeks ap art, are op tim al to protect layers for
the whole p rod u ctio n period; on e d ose alone is partially protective.
Fowl Cholera
I Fowl ch olera (F C ) is a c o n ta g io u s d ise a se o f d o m e stic an d wild
b ird s, d u e to a G ram -n eg ativ e b a cte riu m , Pasteur ella mu Ito ci da.
: T h e cou rse o f the d isease is usually acute, sep ticem ic, w ith high
m ortality, b u t ch ronic form s often occur. Five serogro u p s, nam ed
A , B, D , E, F, have been recognized, on the basis o f their capsu lar
antigens; serovars A and D are the m o st c o m m o n in poultry.
P. multocida p ro d u c e s e n d o to x in s, w h ich m ay c o n trib u te to an
increase in virulence.
Epizootiology
All types of bird are susceptible to F C . A m o n g poultry, ducks, geese
an d turkeys are the m o st affected ; in ch ickens, m o rtality usually
occurs during the laying period and in the hot season. T ransm ission
occurs by direct and in direct contact.
Chronically infected birds are considered a m ajor source o f infection,
but m ost species of wild and d om estic anim als and insects can serve
as vectors. D isse m in atio n o f P. multocida w ithin a flock takes place
prim arily th ro u gh oroph aryngeal excretions or d ead carcasses.

In acu te F C , sig n s of in fection are o fte n p re se n t for on ly a few
h ou rs before d eath ; this m ay so m etim es be the first in dication o f
! disease. If signs occur, these are anorexia, ru ffled feathers, dyspnea,
oral m ucou s discharge, diarrhoea, cyanosis o f the com b and wattles.
T h e lesio n s are asso ciated w ith v ascu lar d istu rb an c e : h yperem ia,
w idely d istrib u ted haem orrhages, exudate in cavities, sw ollen liver
w ith m u ltip le sm all focal n ecro tic areas, p n e u m o n ia an d large
am o u n ts o f m u co u s e x u d ate in the in testin e, p articu larly in the
d u o d en u m .
In c h ro n ic F C , w hich m ay follow an acu te stag e or be d u e to
m icro o rgan ism s o f low viru lence, sw elling o f w attles, sin u ses and
jo in ts, trach eal rales and to rtico llis are the m o st c o m m o n signs.
T h e lesio n s are m ostly ab scesses o f w attles, sin u sitis, p n e u m o n ia ,
arthritis.
60
Diagnosis
A p re su m p tiv e d ia g n o s is o f F C c a n be m a d e fro m c lin ic a l
o b se rv a tio n s an d n ecrop sy fin d in g s, c o n firm e d by iso la tio n o f
P. multocida from tissues and exu dates on blood-agar plates.
Control
T reatm ent o f acute form s o f F C is based on the ad m in istratio n o f
antibiotics, such as am oxicillin, ch in olo n es, etc., by injection or in
drinking water.
Diffuse necrotic foci on the liver due to
P. multocida infection
Prevention o f in fection is based on go o d m an ag e m e n t p ractices,

san itation m easures and exclusion of wild birds, rod en ts and other
anim als.
As regards vaccination, com m ercial bacterins are prepared with all
or. usually, only the m ost com m o n serovars (A, D) o f the bacterium ,
inactivated and su sp en d ed in oily em u lsion .
Two in je c tio n s o f v accin e are given, at 8-10 w eeks o f age an d
4-5 weeks later; in turkey breeders; a b o o ster before g oin g into lay
is som etim es advisable.
NECROTIC ENTERITIS
Necrotic Enteritis
N ecrotic enteritis (N E ) is cau sed by Clostridium per/ringens, type A
an d C , which prod u ce a CC toxin, type C also p ro d u c in g a (3-toxin,
which are responsible for intestinal m ucosal necrosis. C. perfringens
can be iso lated readily on b loo d agar at 3 7 C u n d er an a e ro b ic
con d ition s.
Epizootiology
M ost of the reports o f N E have been in 3-5 week old broiler chickens,
reared on the litter; young turkeys may also be affected.
Initial acute phase of intestinal lesions in NE
P re d isp o sin g cau se s are: d ie t (h igh levels o t barley and w h eat),

d am ag e to in testin al m u co sa (fib res, c o c c id io sis), im b a lan ce o f
m icrobial flo ra (selective an tibiotic treatm ent) an d the ban n in g o f
an tib io tic growth p rom oters in m any cou n tries.

Severe depression , reluctance to move, decreased appetite, diarrhea
and m ortality are present.
G ro ss lesions are confined m ainly tc the sm all intestine; the m ucosa
is lined by a loosely-totighdy-adherent veil '.v or green pseudomem brane,
som etim e with h aem orrh ages.
Diagnosis
T h is is b ased on typical gr< ir .d m ic ro sc o p ic le sio n s an d on
isolation o f large num bers
IC*
co n ten t and scrapin gs frorr. :h e
* C- >s^nngens from the intestinal

v 'Tinal wall.
Treatment
O u tb r e a k s o f N E can be
Necrotic phase of intestinal lesions in NE
r*. a d m in is tr a tio n o f m any
an tib io tics, w hich are active

c h in o lo n e s, ty losin ). Pr
'
strep to co ccu s faeciu m ) c a r
62
G r;.m - bacteria (am oxicillin,

: r c i 'U p p le m e n ts (la c to b a c illi,
r :
r r c \ e n t c lo strid ial in fection .
R1EMERELLA ANATIPESTIFER
Poultry' Disease Manual
Riemerella Anatipestifer
i Riemerella anatipestifer (R A ) in fectio n is a c o n ta g io u s d ise a se o f !
1 variou s d o m estic and wild bird s, p articu larly o f d u ck s, geese an d |
turkeys. R A is a G ran v , non m otile rod, 0.3x1-5 m icron s in size;
it grow s well o n b lo o d agar; grow th is so m e tim e stu n te d , so the j
m ed iu m need s su p p le m e n tatio n by yeast extract and c a lf seru m .
I Epizootiology
T he disease occurs worldwide, particularly in countries, w here duck

production is high. A t least 20 different serotypes have been isolated. |
Infection takes place via the respiratory tract or skin w ounds.

T h e sign s m ain ly o b serv ed are o cu lar an d n asal d isch arge, m ild
cou gh in g and sneezing, d iarrh ea an d ataxia.
T h e m ost freq u en t lesion s are fib rin o u s exu d ates involving serou s
I m em b ra n es, p articu larly the p e ric ard iu m , air sac an d p u ru le n t I
: exudates in the nasal sin uses; ch ronic localized infection occurs in ,
the skin an d join ts.
Diagnosis
A lth o u g h a presum ptive d iag n o sis m ay be m ad e from the clinical
signs and lesions, a definitive d iagn osis is m ade by isolation o f R A .
Control
T h e m o st im p o rta n t m ean s o f p reven tion are g o o d biosecu rity, !

m an ag em en t an d health practices.
In activated b a cterin s are still in use, c o n ta in in g m ore than o n e I
serotype, th ose p re d o m in an t in an area.
T re a tm e n t w ith v a rio u s a n tib io tic s is e ffic a c io u s: am o x ic illin ,
en ro flo xacin , etc..
Mycoplasmosis
Mycoplasmas are very small bacteria lacking a cell wall; they are
consequently fragile and do not survive for long in the environment,
outside the host. Some species live inside the cell, and are therefore
not susceptible to attack by antibodies.
Many species of mycoplasmas have been isolated from poultry, but
the most imporcant and pathogenic are M. gallisepticum , M . synoviae
and M. m eleagridis. They require special enriched media for growth,
which is rather slow (3-10 days); addition of penicillin and thallium
acetate inhibits or retards the growth o f contaminants; colonies are
small (0-1-1-0 mm), with a fried egg form and wedge growth in
solid media. Serological and molecular methods are used for species
differentiation.
M ycop lasm a gallisepticum in fe ctio n

M. gallisepiicum (M G ) is the m o st p a th o g e n ic an d e co n o m ically
sig n ific a n t m y co p lasm a in p ou ltry. T h e in fectio n is c o m m o n ly
k n o w n as c h ro n ic re sp ira to ry d ise a se ( C R D ) o f c h ic k e n s an d
infectious sinusitis o f turkeys. However, the m icroorganism has also
been isolated from m any oth er species o f d om estic and w ild birds,
som etim e w ith occu rren ce o f disease.
S in ce M G rarely survives for m ore th en 2-3 days o u tsid e th e host,

th e p resen c e o f carrier b ird s is e sse n tia l fo r p e rsiste n ce o f the
Characteristic airsaccuiitis due to
M. gallisepticum, often superinfected with . coli
infection in a farm. Infection may spread via fom ites (airborne du st

and droplets, feathers) and personn el (nose, hair). H orizontal spread
increases with the increase in p ou ltry density in sid e the flock an d
a m o n g flocks.
V ertical tran sovarian tran sm issio n is very im portan t- T h e h ighest
rates o f egg tran sm issio n in b reed ers have been fo u n d d u rin g the
acute phase o f disease, b u t this con tin u es, at a lower level, m ore or
less for the entire life o f the bird s; as a c o n seq u en ce, the progeny
are infected. H atch ability is variable and m any un hatched em bryos
show airsac c u litis. T h e in cu b atio n p e rio d o f the d ise a se is very
variable, from 4-5 days to several weeks, d epen din g o n the virulence
o f the M G strain, the presence o f virus and other bacterial infections
and stress factors (environm ent, season, onset of egg production, etc.).
In ch ick en s the m o st c h arac te ristic sig n s o f in fe c tio n are n asal

d isch arge, trach eal rales, sneezing, d ysp n ea, red u ced feed in take
an d higher feed con version rate. In b ro iler flocks, m ost ou tbreaks
o ccu r at 4-6 w eeks o f age; sign s are o fte n m ore m arked th an in
b reed ers an d layers and m ore severe, d u e to freq u en t co n cu rren t
b a cte rial in fe c tio n s, m ain ly c o lib a c illo sis. In layin g flo c k s, egg
p ro d u c tio n declin es or rem ain s at a lower level.
In turkeys, m ore severe respiratory signs d evelop than in chickens,
in clu d in g m ore or less evident sin u sitis.
M o r b id ity is h ig h , m o r ta lit y v e ry v a r ia b le , d e p e n d in g on
com plication s d u e to oth er m icroorganism s. F u rth er losses are due
to retarded grow th, d ow n grad in g o f carcasses an d c o n d e m n atio n .
65
G ross lesions consist prim arily o f catarrhal exudates in nasal passages,

trachea, bronchi an d air sacs: sinusitis is m ost p rom in en t in turkeys.
In severe a n d c o m p lic a te d o u tb re a k s, fib r in o u s a ir sa c c u litis,
perihepatitis and pericarditis are present, resulting in high m ortality
a n d co n d e m n atio n o f birds at the p rocessin g plant.
A presum ptive diagnosis is based on reported sym ptom s and lesions.

Isolation o f the organism from the respiratory tract in special broth
a n d agar-m edia or in e m b ry o n a te d eggs or d e te c tio n o f relative
Commercial broilers with conjunctivitis and
increased lacrimation due to M. gallisepticum
D N A using P C R kits are a con firm ation . Serological tests (SAT, HI

an d E L IS A , see p age 83) are u sefu l fo r flo c k m o n ito rin g in M G
control p ro g ram s an d as an aid in d iagn osis, w hen an in fection is
su spected.
S in c e M G c an b e egg-tran sm itted , m ain ta in in g M G -free p aren t

flo ck s sh o u ld be ideal, alth ou gh this c o n d itio n is n o t very easy to
achieve, particularly in densely populated areas with different poultry
types and farm s. A dequ ate an d severe biosecurity m easures, to avoid
in troduction o f the organism to a flock, m u st be taken. M G control
p ro gram s have b e e n a d o p te d w ith great su ccess in ch ick en s and
turkeys, prim ary and m u ltiplier breeders, reared in strict isolation.
G iv en the c o n sid e ra b le d iffic u ltie s in k e e p in g flo c k s M G -free,
a p p ro p ria te an tib io tic th erapy an d v a c c in a tio n m ay be u sed to
reduce th e d am age due to infection, particularly egg tran sm issio n ,
b u t can n o t prod u ce the com plete elim ination ot the organism from
all bird s in an in fected flock. M G h as been show n to be sensitive
to several an tim icrob ials, in clu d in g m acrolid es, flu o ro q u in o lo n e s
an d others. They are able to reduce the severity o f clinical signs and
lesions and egg transm ission, but they often need to be adm inistered
again and again. Egg injection or dippin g have been used to introduce
antim icrobials into hatching eggs, limiting, bu t usually not completely
elim in atin g the organ ism .
A s regards v a cc in atio n , in activ ated an d live atte n u a te d vaccin es
have lon g been used in b reed ers, bu t m ostly in m u ltiaged layers,
Commercial broilers with conjunctivitis and
increased lacrimation due to M. gallisepticum
with fairly g o o d results.

In activ ated v accin e s or b a cte rin s in oily e m u lsio n , c o n ta in in g a
high titre o f antigen, m ust be injected twice, at least 4-5 weeks apart,
for optim al protection , also again st egg tran sm issio n o f organ ism s.
Live attenuated vaccines are ad m in istered by spray and by eye-drop,
in 6-8 w eek old chickens. C u rre n tly av ailab le M G vaccin es have
n o t been m uch used in turkeys.
66
Mycoplasma Synoviae Infection

\ Mycoplasma synoviae (M S) prim arily in d u ces in fla m m a tio n o f the
synovial m em b ran es o f jo in ts an d te n d o n sh eath s (synovitis), b u t
frequently there is also respiratory involvem ent, m ostly in chickens
and turkeys.
In m any respects, the sp re ad o f M S ap p e ars to be very sim ilar to

that o f M G . T ran sm issio n occu rs horizontally, via the respiratory
tract and vertically, th ro u gh the ovary and oviduct. T h e in cu b ation
perio d is very variable, from 6 to 7 days, m ostly in chicks in fected
vertically by egg tran sm issio n , to several weeks.
Tarsal-metatarsal joint synovia of a) normal;

b) M. synoviae affected chicken
T he m ost frequent signs in chickens are pale com b, retarded growth,

lam eness, sw elling o f jo in ts (m ainly h ock and foot pads) an d breast
b liste rs; so m e tim e s, m o stly in b ro ile rs, re sp ira to ry sy m p to m s.
Recovery, w hen it occurs, is very slow, b u t synovitis may p ersist for
the en tire lifetim e. Egg p ro d u c tio n m ay be m ore or less affected,
p a rtic u la rly jf th e d ise a se o c c u rs w hen th e b ird s go in to lay.
S om etim es the in fection course is asym ptom atic an d revealed only
by sero lo g ical tests. M o rb id ity is very variab le, from 2 % to m ore
th an 5 0 % ; likewise, m ortality can be from less then 1% up to 10%.
G r o ss le sio n s in clu d e th ic k e n in g o f sy n ovial m e m b ra n e s w ith
viscous, creamy to caseous exudates, with spleno- and hepatom egaly;
in respiratory form s, airsaccu iitis m a y b e present.
In turkeys, M S gen erally cau ses sign s and lesio n s very sim ila r to
those o f chickens.
A presum ptive diagn osis is based o n characteristic signs an d lesions.

C o n firm a tio n may be o b tain e d by iso latio n and id e n tificatio n o f
M S , m ore fastid io u s th an M G to grow, o r by d etectio n o f relative
D N A , u sin g PG R . F o r an an tib o d y survey, the SA T, HI o r E L IS A
tests are used.
S in ce M S is egg-transm itted, the sam e rules ad o p ted for M G m ust

be ap p lied . M S is sensitive to several an tib iotics; their use m ay be
m ore extensive than in M G infections.
A n inactivated b acte ria in oily em u lsion is com m ercially available;
two in jection s, at least 4-5 weeks apart, ate needed to o b tain g o o d
im m unity.
Tarsal-metatarsal synovia affected

by M. synoviae
68
Mycoplasma Meleagridis Infection

Mycoplasma meleagridis (M M ) is a sp ecific p ath o ge n o f turkeys, in
w h ich it cau se s a irsa c c u iitis in th e p ro g e n y an d p o o r grow th
perform ance.
M M is vertically tran sm itte d by the egg; the o rg a n ism h as b een

recovered from variou s sites in the oviduct, b u t rarely in the ovary.
H o riz o n ta l tra n sm issio n m ay o c c u r at any age, d irectly by th e
airb o rn e route or indirectly by artificial in se m in atio n , sexin g an d
other practices.
In sp ite o f m ore o r less severe airsaccu iitis, resp iratory sign s are
rarely ob served in egg-borne infected p ou lts. In ad u lt turkeys, the j
course o f infection is m ostly silent. Air-sac lesion s are characterized
by thickening o f walls w ith yellowish and caseous exudate. Skeleton
lesion s m ay som etim es be present.
A presum ptive d iag n o sis is based particularly o n exudative lesion s

o f the air-sacs, alth o u g h th ese sym pto m s are n o t always p resen t.
D ia g n o s tic c o n fir m a t io n m ay be o b t a in e d by is o la tio n a n d
id en tifica tio n o f M M o r by d e te c tio n o f the relative D N A u sin g
P C R . For an in vestigation o f an tib o d ie s, the SA T, H I and E L IS A
tests are used.
1
M a n a g e m e n t p ro ced u res are the sam e as th ose ap p lied for oth e r I
m yco plasm as, aim ed at e rad icatio n . M M is sensitive to the sam e
a n tib io tic s u sed a g a in st o th e r m y c o p la sm a s. V ac c in e s are n o t
available.
C O C C ID IO S IS
Coccidiosis
C o c c id io sis is an in testin al disease o f alm o st universal im p ortan ce
in p ou ltry p ro d u ctio n , cau sed by in tracellu lar p roto zoan parasites
o f th e g e n u s Eim eria. G e n e ra lly th e c lin ic a l d ise a se is ra th e r
u n c o m m o n in in te n siv e ly -re a re d p o u ltr y d u e to th e u su a l
ad m in istration o f coccidiostats or vaccines; nevertheless, subclinical
and d etrim en tal in fection s are norm ally detectable.
C occid ia are species-specific an d have a com plex life cycle, with one
p art in sid e an d on e p art o u tsid e the body, w here m a tu ra tio n o f
oocysts takes place, d ep e n d in g o n environ m ental tem perature an d
h u m id ity. F ro m ju st o n e oocy st o f E. tenella, afte r 3 rep licatio n
Lesions caused by E tenella in ceca
cycles (as schizonts and m erozoites, the invasive form o f the parasite)
an d on e sex u al stage in cells o f the in testin al m u co sa (w ith the
u n io n o f m icro-gam etocytes an d m acro-gam etocytes), on e chicken
can elim inate ab ou t 4 0 0 ,0 0 0 oocysts. T h is intensive elim in ation o f
oocysts in a lim ited area with high num bers o f chickens, as norm ally
occurs in an intensively-reared flock, is the reason for the im portance
o f the d isease in m od ern pou ltry p ro d u ctio n .
In c h ic k e n s, seven sp e c ie s o f E im e ria have b e e n id e n t ifie d :
E. acervulina, brunetti, m axim a, mitis, necatrix, praecox and tenella.
Each species differs from the others in pathogenicity, im m unogenicitv
and the intestinal replication site, with different degrees o f intestinal
le sio n s. T h e m o st im p o rta n t sp e c ie s fro m an e c o n o m ic p o in t
o f view in chickens are E. acervulina, m axim a, tenella an d necatrix
(th e latter c a u sin g the c h a ra c te ristic c o c c id io sis in 8 w eek-old
chickens). E. precox an d E. mitis can cau se loss o f p erfo rm an ce, in
exp erim en tal trials only, w h en chicks are ch allen g ed w ith a very
high d o sag e o f oocysts.
In turkeys, where coccid io sis is also co m m o n , b u t less sp ectacu lar
and im p ortan t then in chickens, five species are the m ost im portan t
E. adenoides, gallopavonis, meleagridis, meleagrimitis, dispersa.
Haemorrhagic lesions caused by E. tenella

in the caecum
In geese, E. truncata an d E. anseris are co m m o n ; in ducks, Tizzeria

perniciosa.
Epizootiology
O u tb reak s o f d isease in chickens are co m m o n at 3-6 weeks o f age.
S ev eral o u tb re ak s are p o ssib le in the sam e flo c k w ith d iffe re n t
species involved in each, becau se o f no cross-protective im m unity.
70
B reed er an d layer pu llets are affected at the sam e age as bro ilers
an d th en at 9-18 w eeks by E. necatnx. C o c c id io sis rarely occu rs in
adult birds, probably due also to gradu al im m u n ization at a young
age. T h e n atural m ean s o f tran sm issio n is by in gestio n o f oocysts,
which are very resistant in the environm ent and also to disinfectants.
Symptoms and lesions

C o c c id ial in fection s may be classified as:
- clinical coccidiosis, characterized by m ore or less slight m orbidity,
Lesions caused by E. aceivulina in the small
intestine
an d m ortality, d ia rrh o e a o r b lo o d y faeces, as well as by lo ss o f

perform ance;
- su b clin ical co ccid io sis, in w hich clinical sign s o f d isease are not
very evident, b u t c au se a re d u c tio n in w eight gain an d the feed
conversion rate;
- asym ptom atic coccidiosis, which d oes n ot cause significant adverse
effects in the host.
L esio n s are located in d iffere n t p arts o f the in testin e, d e p e n d in g
on the strain o f Eimeria involved, with d ifferen t lesions.
Diagnosis
D iagnosis is based on the localization and nature o f the lesions and
on a m icroscopic exam in atio n o f m ucosal scrapings, m easurin g the
size an d sh ap e of oocysts and scorin g the lesion s.
Control
T h e response o f the im m u n e system o f poultry to coccidia plays an
im portant role in controlling the disease. T he host im m une response
in fact, constandy influences the dynam ics o f the coccidial population
in flocks. Both celbm ediated and hum oral im m unity are activated,
b u t the m u co sal ce lb m ed iate d re sp o n se seem s to be o f p rim ary
im p o rta n c e in re d u c in g the cap acity o f c o c c id ia to rep licate in
intestinal cells.
For this reaso n , an tic o c c id ia l d ru gs, n o t very selective as regards
c o ccid ial rep licatio n , as well as live vaccin es, have been used for
long-term control o f the disease. Synthetic chem icals, particularly
if highly e ffica cio u s, w hen ad m in iste re d to p reven t co c c id io sis,
Lesions caused by E. maxima in the small
intestine
rap idly select n atu rally -resista n t stra in s, n o t allo w in g sen sitiv e
coccidia to replicate and blockin g natural com petition between the
d ifferen t strain s present in flocks.
Io n o p h o te s, b ein g co ccid io static, and con seq u en tly less effective,
allow the im m u n e system to respond; the lower degree o f selection
tow ards resistan t strain s p erm its their use for a longer tim e before
inducing resistance (m onensin, narasin, salinom ycin, sem duram icin

and lasalocid).
T h e use o f live v accin e m ak es p o ssib le a sh ift in the c o c c id ia l
p o p u la tio n from d ru g resistant to drug sensitive an d a progressive
su b stitu tio n o f wild w ith v a cc in e ' atte n u ate d oo cy sts, red u cin g
in testin al stress, w ith an in crease in p erfo rm an ce o f the chickens.
T h e early vaccines were prod u ced with wild strains o f coccid ia on
the basis that precociou s exposure to a sm all d ose o f oocysts is able
to in d u ce a p ro tectin g im m u n e resp on se; however, n o n -u n iform
a d m in istratio n o f these vaccines m igh t in duce clinical coccidiosis.
T h e a d o p tio n o f su ccessfu l m eth od s o f coccid ial atten u atio n , has
p erm itted the o b tain in g o f protective vaccines with a large m argin
o f safety. A tte n u atio n is p erform ed by;
- selecting for p recocio u sn ess, in w hich a red u ction in the life cycle
tim e is co m b in e d with low invasiveness;
- su ccessiv e p a ssa g e s in ch ick en e m b ry o s; th is latte r m e th o d is
ap p lied only with certain species o f Eimeria an d stu d ies reveal that
the safety and stability' o f atten u ation w ould be com parable to that
o f p recocio u s strains.
U sin g live vaccines, only one adm inistration is sufficient for com plete
im m unity, becau se chicks repeatedly take vaccinal oocysts from the
; litter.
E. brunetti
r -j
js
Vaccine adm inistration system

T h e p u rp o se o f the use o f v accin es is to p reven t o r red u ce the
econ om ic d am age cau sed by in fectious diseases in treated bird s or
to p reven t the vertical tran sm issio n o f in fection to the progeny.
H y p erim m u n izatio n m akes it p o ssib le to rein force resistan ce to
infection, but also to maximize maternally-derived antibodies, which
: will p rotect the progeny fo r the first 2-3 w eeks o f life, w hen birds
are m ost su sceptible.
| V accin es are divided into two m ain classes, an d are p rep ared with;
- live m icroorganism s, naturally apathogenic, m ore or less attenuated
in ch icken em bryos or in tissue cultures, deriving from d ifferen t
species, with varying am o u n ts o f an tigen per d ose, b u t involving
rep lication o f the vaccinal agen t in the host; a seco n d type o f live
vaccine is em erging with the developm ent o f genetically engineered
live viruses and bacteria, vectored vaccines or gene-deletion m utants
o f pathogen ic m icroorganism s and, m ore recently, D N A vaccines;
- inactivated m icroorganisms, mostly using formalin or (3-prop iolactone
as inactivant, com b in ed with an ad ju van t (alu m in iu m hydroxide,
mineral oil, others), obviously containing large am ou n ts o f antigen,
there being no rep lication in the host.
, T h e ap p licatio n tech n iq u es are su bd ivid ed into: m ass vaccin ation
' (spray, ae ro so l an d d rin k in g w ater) an d in d iv id u a l v a cc in atio n
(eye-nose-drop, wing-web, in tram u scu lar, su b c u tan e o u s o r in-ouo),
in the hatchery o r on the farm .
Vaccination via the drinking water

Tliis is the m ost com m only used vaccination technique in com m ercial
poultry. However, the factors, which affect its success are num erous.
T h e w ater used m u st be clean an d d rin k ab le, as free as p o ssib le
from b acteria, o rg a n ic su b sta n c e s, m etal ions, ch lo rin e and any
d isin fectan ts, which may neutralize vaccinal m icroorgan ism s, with .
a pH between 5.5 and 7.5. Dried skim m ed milk (5 to 8 g/litre) should j
be added to the water, to neutralize at least part o f the contam inants. 1
B ir d s s h o u ld be m a d e th irsty , th u s d e p r iv e d o f w a te r fo r
1 to 2 h ou rs, d e p e n d in g on the clim atic c o n d itio n s; c o m p e titio n
trz
" ' drinking water
am ong the birds m ust be avoided during vaccinal water consum ption,
w hich m u st take p lace over a b o u t 2 h ou rs. T h e vo lu m e o f w ater
for vaccin atio n sh o u ld be a b o u t 1 /5 o f the p rev io u s d ay s w ater
75
j c o n su m p tio n . D rin k e rs m u st w ork correctly. Finally, the farm er

sh o u ld carefully m o n ito r the poultry h ou se, to check th at all birds
. have en o u gh to drink.
Vaccination by spray
T h e m eth od is d esig n ed to m im ic eye-drop vaccin ation , b u t with
the advantage o f a m u ch lower cost o f ad m in istration . It allows the
vaccine to be b ro u g h t into con tact with the eyes, F latd erian glands
an d u p p er respiratory tract. T h e d ro p lets, generated by a sprayer,
m u st have a size o f a b o u t 100-150 m icro n s. D ro p le ts, w hich are
too fine, m ust be avoided, as they penetrate deeply into the respiratory
tract (bron ch i an d lungs) and cou ld in du ce post-vaccinal reactions
with viruses wrhich are n ot com pletely ap ath o gen ic, particularly in
cases o f N D V , at least for the first vaccin ation .
Manual vaccination by spray
Spray vaccination in the hatchery

T h is is usually d o n e u sin g a spray cab in et, w hich sprays b o xe s o f
one-day-old ch icks m ovin g o n a conveyor b e lt w ith a c o n tro lle d
sp e e d . N D V an d IB V v a c c in e s are n o rm a lly a d m in iste re d in
30-40 m l o f d istille d or p u re w ater p er 100 ch ick s; c o c c id io sis
v a cc in e can also be a d m in iste re d . S u b se q u e n tly , ch ick s p reen
them selves or each other, takin g ad d itio n al vaccine.
Vaccination by spray machine in hatchery
S p ra y v a c c in a tio n o n t h e fa r m
T h is is don e using a sprayer equipped with a plastic tank containing
5 to 10 litres and a spraying arm o f ab ou t 70 cm with 1 or 2 nozzles.
T h e quantity o f w ater is 50-80 m l per 100 chicks, the droplets m ust
have a size o f 100-150 m icrons, som etim es an d w ith som e vaccines
from 10 to 100 m icrons. For good vaccination, the following practice
is necessaTy-: grou pin g the birds together, turning o ff ventilation and
heating systems, reducing the light intensity, spraying about 30-40 cm
above the h e ad s o f the b ird s, w alking slowly th ro u gh the h o u se
(exposure for 5-7 seconds) and, better still, repeating the operation ,
fin ally resto rin g the en v iro n m en tal c o n d itio n s after 15 m in u tes.
76
Vaccination by oculonasal route
T h e m ethod is highly effective, bur labour-intensive and expensive; |

it is often used to vaccin ate ag ain st laryngotracheitis. T h e vaccine
is r e c o n s t itu te d in 3 0 -3 5 m l o f d ilu e n t p er 1 0 0 d o s e s a n d
ad m in istered at a d ose o f 1-2 d ro p s per bird, w ithout tou ch in g the
eyes o r n ates; it is im p o rta n t to w ait a few se c o n d s to allow th e
vaccine to be ab sorbed .
Vaccination by oculonasal route
V a c c in a tio n b y w in g - w e b - s t a b
T h is m ethod is used to im m unize again st fowl pox. T h e vaccine is
re c o n stitu te d in 10 m l o f ste rile d ilu e n t p e r 1 0 0 0 d o s e s an d
ad m in istered u sin g two ap p lic atio n tools: a stick with on e or two
g ro o v e d n eed les, w hich are p rev io u sly d ip p e d in v accin e o r an
autom atic so-called G ra n t inoculator, with a self-contained reservoir
for the vaccine. Pierce the wing-web in a d efeathered area.
Im m unization of birds is indicated by a local swelling o f skin at the
pierced site, checked after 8 days.
Vaccination by wing-web-stab
| T h is m e th o d is p e rfo rm e d in the h atch ery d u r in g tra n sfe r o f

in cu b atin g eggs from the setter to the hatch er, at the 18th day of j
incubation. T h e dose of vaccine per egg is 0.0 5 - 0.1 ml, inoculated
th ro u gh a hole on the large end o f the egg, in to the am n io tic sac
or preferably in to the em bryo. A n au to m ated egg in jection system
| is used; on e m achine with 3 p eop le norm ally in oculates 20-30.000
eggs per hour. Sin ce the in o cu latio n leaves a hole in the egg-shell,
fo r the la st few days p r io r to h atc h in g , b a c te r ia l an d fu n g al
| c o n ta m in atio n s are p ossib le; con seq u en tly , the hatchery m u st be
In oi/o vaccination at 18 days of incubation
k e p t well san itize d . T h is te c h n iq u e is a p p lie d p a rticu larly fo r

vaccin ation again st M a re k s d isease with H V T , m ostly in broilers,
b u t trials with oth er vaccines have been con d u cted .
77
Vaccination by injection
T h e injection is accom plished by the su bcu tan eou s or intram uscular
route, k is used for ad m in istration o f certain live vaccines, su ch as
th o se a g a in st M a re k s d ise a se (one-day-old chicks) an d reovirusinduced arthritis (1- to 6-dayold chicks), but also o f many inactivated
vaccines at any age, m ostly after 4 weeks o f age or before going into
lay. Injection can be perform ed by han d, u sin g au to m atic syringes,
o r with au to m a tic m ach in es. T h e precision o f the delivered d ose
m u st be checked befo re an d d u rin g the o p e ra tio n s. N e e d le s are
selected depending on the type o f vaccine, normally 1x15 mm size or less.
Subcutaneous vaccination against
Marek's disease
Su b cu tan eo u s vaccination is m ostly perform ed into the base o f the

neck, this being the clean est area o f the bird, in sertin g the needle
tow ards the body o f the bird. Intram uscular vaccination is generally
perform ed into the breast, leg or wing m uscles, keepin g the needle
at an angle o f 4 5 . It is ad visable to keep the bottle o f inactivated
vaccin e for at least h a lf a day at roo m tem p eratu re (over 2 0 C ),
to im p rove the flu id ity o f the vaccin e, p articu larly if it is in oily
em u lsion .
If the o p erato r sh o u ld accidentally prick h im se lf with the needle,
a d octor m ust be con su lted , particularly in case o f vaccines in oily
em u lsio n , which, even in sm all am o u n ts, cou ld cause severe local
reactions.
Subcutaneous vaccination with inactivated

vaccine
78
Factors influencing vaccination

M an y facto rs can h ave a c o n sid e ra b le im p act o n the su c c e ss o f
v a c c in a tio n ; they have to be tak e n in to g re at c o n sid e ra tio n by
veterinarian s an d farm ers.
Factors related to the conditions of farms

E x is tin g p a t h o lo g ic a l c o n d it io n s d u e t o in f e c tio u s a g e n ts
(v iru s , b a c te ria e tc .).
T h e p re se n c e in th e b ird s o f v iru se s w h ich in d u c e im m u n o
su p p ressio n , particularly C 1A V an d IB D V , have a great in fluen ce
o n v a c c in a tio n re su lts. V accin atio n o f p a re n ts may p ro te ct th e
p rogeny in the p erio d o f the greatest su scep tibility to the d iseases
m en tio n ed an d also to others.
T h e p resen ce o f bacterial in fection s an d , con seq u en tly , the need
for an tib acterial treatm en ts, can also interfere w ith the im m u n e
response to vaccination. In such case, the im m u n e system , focusing
o n che in fectiou s p ath ology, will be less available for the im m u n e
reaction to the vaccine or the vaccinal reaction may w orsen the preexistin g in fection . T h e sam e c o n d itio n ap p lies to clin ical or subclinical coccid iosis. T h erefore, u n d er su ch con d ition s, vaccination
sh o u ld be p o stp o n e d , if possible, fo r 4 to 6 days.
S tre s s
A m o n g the v ario u s effects o f stress o n the o rg a n ism there is also
im m unodepression ; this seem s to be due to increased glycocorticoids
release, which in hibits the synthesis o f m any cytokines involved in
im m u n e processes. D eb eak in g , tran sp o rt, excessive tem peratu re,
overcrow ding, start o f laying, etc. m ay be stressin g factors. Possible
an ti-stress th erap y , su c h as v ita m in c o m p le x e s, to g e th e r w ith
im p rovem en t in hygienic m easures, can h elp to redu ce the stress.
M y c o to x ic o s is
M any m ycotoxins can induce an im m u n osu p p ressive effect, m ostly
a fla to x in s an d o c h ra to x in s. T h e effect is, in p art, e x p la in e d by
partial atrophy o f lym phoid tissues, su ch as the bu rsa o f Fabriciu s,
thym us an d sp leen . T h e c o n tro l o f m y co to x ico sis is, ob vio u sly ,
j b ased o n p rev en tin g fu n gal d e v e lo p m e n t in feed s, also a d d in g
a n t if u n g a l a g e n t s. T h e tr e a t m e n t o f b ir d s s u f f e r in g fro m
m y co to xico sis c o n sists o f h e p ato p ro te c tan ts (m any am in o acids)
an d anti-oxidants, w hich can be qu ite expensive.
F a c to rs a s s o c ia te d with vaccination
H o s t fa c to r.
T h e health status, with particular attention to intercurrent subclinical
o r c h ro n ic in fe c tio n s (M ^coptasmas, E. co!i, etc.) sh o u ld be well
con trolled before and after vaccine ad m in istration , taking the d u e
p re c a u tio n s. T h e e n v iro n m en tal c o n d itio n s sh o u ld be the best
possible.
V a c c in e fa c to r
| T h e vaccin e m u st be as safe as p o ssib le, b u t at the sam e tim e be
; im m u n ogen ic, su ited to the ep id em iological situ atio n (e.g. IB D V ,
m ore or less invasive, d e p e n d in g o n the virulence o f the w ild virus ,
present in the field). T h e tim e o f vaccination should also be carefully
p r o g r a m m e d , d e p e n d in g o n th e e p id e m io lo g ic a l s it u a tio n .
M o st o f vaccines m u st be stored in a refrigerator at 2 -8C until use
| and tran sported u n d er cool con d ition s. T h e n u m ber o f d oses m ust
c o rre sp o n d to the n u m b e r o f b ird s to be v accin ated , never less.
I H u m a n fa c to r
T h e tech n iq u es used to ad m in iste r vaccines m u st be as sim p le as
p o ssib le, b u t the in stru ctio n s m u st be u n d e rsto o d and follow ed
correctly by the w ork ing staff. T h e m ost com plex tech n iq u e is the
on e regard in g p re p a ra tio n an d ad m in istra tio n o f c e lb asso c iate d
M a re k s disease vaccine, due to the fragility o f the cells d u rin g the
various o p eratio n s, for w hich a con troller is advisable.
T h e vaccin atio n o p e ratio n s, particu larly in large farm s, m u st be
p rogram m ed with care and sh o u ld be perform ed in the m orn in g,
p articu larly in h o t se a so n s an d co u n trie s. T h e e q u ip m e n t used
m u st be checked an d prep ared the day before. M o n ito rin g o f the
flock b efo re an d after vaccin ation is very im p o rtan t; the result o f
th e im m u n e r e sp o n se m u st th en be v a lid a te d by s u b s e q u e n t
serological tests.
Diagnosis of disease
j Clinical, necroscopk and laboratory analysis
. T h e goal o f th ese series o f analyses is to d e term in e as quickly as
p o ssib le the cau se o f d isease, in ord er to in terven e in g o o d tim e
with ap p rop riate th erapeu tic an d hygienic m easures.
C ase h isto ry
a) species, breed, age o f birds;
I b) m an ag em en t factors: ven tilation, feedin g, w atering system , feed
c o n su m p tio n , b ro o d in g and rearing p roced u res;
c) evolution o f p erform an ce: grow th or laying curve;
d) previous vaccinations and treatm ent schedules (nature o f vaccines
! an d d ru gs, a d m in istratio n rou te, d osage, d u ra tio n o f treatm ent);
j e) previou s p ath o lo g ic al h istory an d d ate w hen p ro b le m started,
type an d d u ra tio n o f sign s o b serv ed a n d / o r rem em b ered in the
flock, any lesion s ob served , evo lu tion o f m orbidity an d m ortality.
C lin ic a l e x a m in a tio n : p resen ce o f cu tan e o u s lesio n s, respiratory
1 sym p to m s (sneezing, rales, n asal d isch arg e, sin u s sw elling, etc.),
enteric sym ptom s (appearance an d colou r o f d iarrh oea and faeces),
j nervous an d lo c o m o to r sy m pto m s (w eakness, leg in co o rd in atio n ,
trem or, spastic or flaccid paralysis, b lin d n e ss, etc.).
N e c ro p sy : a post-m ortem e x am in a tio n is p erfo rm e d to h ighlight
lesion s w hich are m ore or less ch aracteristic or p ath o g n o m o n ic o f
certain diseases (for exam ple, sw elling, o e d em a and haem orrh ages
o f the bursa o f Fabricius, d u e to IBD V ; h aem orrh agic typhlitis d u e
to coccidiosis; h aem orrh agic and necrotic lesion s in the gut due to
N e w c a stle d ise a s e ) a n d to ta k e s a m p le s fo r p a r a s it o lo g ic a l,
bacteriological, virological an d h istop ath ological analysis.
N ecropsy m ust be p erform ed on sick birds, h u m an ely su p pressed ,
o r on freshly-dead birds. T h e sam p les o f o rgan s o r tissu es, sw abs
o r sm ears for fu rth e r e x a m in a tio n m u st be tak e n u sin g asep tic
procedures, in order n ot to contam inate the sam ples with extraneous
m icroo rgan ism s.
T h e tran sp o rt o f sam p les o f bird s or p art o f this m u st be d o n e in
such a w7ay as to prevent any co n ta m in atio n o f p erson s involved in
tran sp o rtin g an d receiving, an d to preserve the m aterial quality o f
the sam p les (refrigeration , freezing, 10% fo rm alin fixatio n , etc.);
transport boxes, norm ally o f plastic, m ust be labelled as pathological
81
m a te r ia l , in te n d e d fo r e x a m in a tio n an d a c c o m p a n ie d by an
inform ation sheet; inform ation m ust be as com prehensive as possible,
in ord er to m ake die d iag n o sis easier.
T h e site for necropsy and exam in atio n can be a disin fectable room
at the farm o r n ear to the carcass d isp o sa l site, b u t the necropsy
room o f a fitted o u t an d w ell-equipped lab o rato ry is preferable.
A ny e q u ip m e n t o r c lo th in g m u st b e c le a n e d an d d isin fe c te d ,
sterilized oi\ if d isp o sab le, in cinerated.
Sampling methods
B lo o d sam p le s: tubes for seru m based tests (serology) or tubes with
an an ticoagulan t for biochem ical investigations (turn the tube gently
several tim es) are used. B lood is taken individually by decap itation ,
(very young chicks) o r intracardiac puncture, or wing-vein sam p lin g
or at slaughter.
Sam p les m ust be ab o u t 2-3 m l in volum e, in plastic tubes, clean or
sterile an d sealing. S to ra g e for less than 4 8 h o u rs can be at 4-8C ,
for a lon ger period at - 2 0 C , in the later cases only for the serum ,
Wing-vein blood sampling
after sep aratio n from clotted b lo o d .

Tw o sets o f sera from th e sam e bird s, collected d u rin g the acu te
p h ase (at the start o f the d isease) an d con v alescen t p h ase (after at
least 2-3 weeks) o f the d isease, are tested to h igh ligh t the increase
in an tib o d y titres again st a path o gen su sp ected o f b ein g the cause
o f rhe disease and to m on itor re sp on ses to vaccin ation s.
Organ or tissue samples

F o r p a ra sito lo g ic a l e x a m in a tio n : sc rap in g o f in testin al m u co sa,
faeces (coccidia, w orm eggs, protozoa) and investigation o f protozoa
m u st be d o n e im m ediately after sam plin g.
F o r b a cte rial iso la tio n : large pieces o f organ s (spleen , liver, lung,
intestine, etc.) are placed in the transport m edium and then refrigerated
or frozen;
F o r viru s iso latio n : sm all pieces (2x1 cm) are placed in preservative,
possibly su p p lem en ted w ith antibiotics, preserved in 5 0 % glycerol
or frozen.
T issu e sm e a rs fo r b a c te r io lo g ic a l, v iro lo g ic a l an d h isto lo g ic a l
exam in atio n m u st be dried an d fixed in methyl alcoh ol to prevent
deterio ration .
T h e sam ples m u st be sent to the laboratory as so o n as p ossible and
correctly stored, refrigerated or frozen, d ep e n d in g on the tran spo rt
tim e.
82
Bacteriological examinations
T h ese are d on e to identify the presence o f p ath ogen ic bacteria an d
to test their sensitivity to an tibacterial dru gs (antibiogram s).
They include: direct m icroscopic observation o f bacteria or protozoans
in the tissues (fresh or stained); isolation in different selective culture
m ed ia m ain tain e d at 3 7 C for a p erio d from 24 h o u rs to several
days, d e p e n d in g on the m ic ro o rg a n ism ; b io c h e m ic a l p ro file s;
an tibiogram s.
SAT: positivity is shown by a visible dumping
of the stained antigen
Serology
In fection by a p ath o g e n n o rm ally in d u ces p ro d u c tio n o f specific
an tib o d ies.
Serological tests are used to highlight these antibodies for diagnostic
p u rp o ses, epizoological m o n ito rin g a n d to c o n firm a d iag n o sis o f
d isease, in ad d itio n to m o n ito rin g and q u an tify in g the an tib o d y
resp on se o f bird s to vaccin ation .
T h e serological tests m o st com m o n ly u sed are:
Agglutination tests, slide (SA T) or tube (TAT) agglutination, to detect
an tibacterial an tib o d ie s (Salmonella spp., Mycoplasmas)-,
GelPrecipitation test or agar-gel im m u n o d iffu sio n test (A G ID ), where
the antigen and sera are placed in two ad jacen t wells previously cut
in to the agar; a fte r 24-48 h o u r s, a p r e c ip ita tio n lin e o f th e
an tigen /an tib o d y com plex appears in the case o f a positive reaction;
this test is used for IBV, IBD V , In flu en za, etc.;
AGD test with feather tips: precipitation line

Haemagglutination4nhibition (HI) test, based on the ability o f certain

v iru ses o r b acte ria to agg lu tin ate red b lo o d cells (R B C ); a fixed
quantity o f antigen (4-8 H A units) is placed in contact with successive
dilutions (log2) o f serum to be tested in the well o f a plastic m icroplate;
after 30 m inutes, a 0 .5 % su sp en sio n o f R B C is added, w ith shaking
a n d after a fu rth e r 3 0 m in u te s th e test is read : se d im e n ta tio n
in dicates a positive H I reaction , agglu tin atio n in d icates a negative
reaction; this test is used for N D , E D S, IB, influenza, mycoplasmas, etc.
Enzyme linked immunosorbent assay (E L ISA ). M any types o f E L IS A
tests have b een d evelop ed . T h e sim p le st form (in d irect E L IS A ) is
based o n th e b in d in g o f an tib o d y to an tig en , previously fixed at
the b o ttom o f a micro-well; antigen-antibody bin d in g is revealed by
a d d itio n o f a n en zym e-labelled an tib o d y an ti-a n tib o d y a n d by
su b seq u en t ad d itio n o f su b strate (crom ogen). T h e su bstrate, after
b ein g hydrolyzed by the enzym e, ch anges colou r; the in tensity o f
the c o lo u r change is p ro p o rtio n a l to th e q u an tity o f a n tib o d ie s
HI test: Hl-positive walls show a tight,

drcumscribed button of unagglutmated,
sedimented erythrocytes
b o u n d to the an tigen ; read in g o f the reaction is p erfo rm ed u sin g

a spectrop h oto m eter (optical density). T h e m eth od is qu ite sim ple,
D IA G N O S IS O F D IS E A S E
rap id an d ot low cost; it allow s a large n u m b er o f seru m sam p les

to be exam in ed an d can be used for nearly all p o u ltry d ise a se s.
O th er E L IS A test are the san d w ich an d the com petitive types.
Polymerase chain reaction (PC R ): this test was extensively introduced
m ore recently in the d iag n o sis o f in fection ; nucleic acid segm en ts
o f a m icroorganism are revealed in a sam ple o f tissue or body fluids
(trachea, in testin e etc.).
Enzyme Linked Immunosorbent Assay (ELISA):

the color change indicates the amount of
captured antigen or antibody
Diagnosis of some most

im portant Poultry Disease
NEWCASTLE DISEASE
Newcastle disease (ND)
Clinical signs:
Respiratory sym ptom s
Enteric sym ptom s
N ervous sym p to m s
Necropsy
Intestinal necrosis ,,
and haemorrhages, v
proventriculus
haemorrhages
M o rta lity
Laboratory
Virus isola tio n fro m
intestine, lung
and brain
YES
NO
NO
C ontrol fo r o th e r diseases:
ILT, IB, M ycoplasm osis, etc.
D iagnosis of ND
Laboratory
Serological tests (H lV
- C ontrol o f vaccination
schedules
- Vaccine type and strain
- C ontrol o f vaccine
a d m in istra tio n
D iagnosis o f ND
YES
NO
C on trol fo r o th e r diseases:
ILT, IB, Mycoplasmosis, e tc
ND Lesions
Nervous sym ptom s
Nervous signs, like twisted neck or torticollis, are characteristic of nervous lesions j
caused by certain neurotropic and mesogenic strains of ND
Gastric and enteric lesions

Haemorrhages /oca/ized in the stomach and intestine are characteristic of ND
Normal stomach in a broiler:

a) proventriculus;
b) gi22ard
Haemorrhages present in the proventriculus

and sometime in the gizzard
Normal intestine
Haemorrhages in the intestine
Necrosis in the intestine
87
IN F E C T IO U S B R O N C H IT IS
Infectious Bronchitis (IB)
Clinical signs:
R espiratory sym ptom s
D iarrhoea
Egg drop
YES
Necropsy
Laboratory
Virus isola tio n fro m
trachea a n d /o r
kidneys
Characteristic lesions in:
- trachea
- kidneys
NO
Y ES
NO
C on trol fo r o th e r diseases:
ILT, ND, Mycoplasmosis, etc.
D iagnosis o f IB
Laboratory
S erological tests
at the tim e and
15-20 days
later
Control of vaccine
adm inistration
Control of vaccine
schedules
Positive
D iagnosis of IB
N egative
ILT, ND, Mycoplasmosis, e tc
IB Lesions
Trachea
Piccure of a normal trachea, compared

with a trachea with IB lesions characterized
by a catarrhal-hemorrhagic exudate
Kidney
In nephropathic infection, swollen and pale kidneys due to interstitial nephritis and 1
distended ureter with urates are present
A) normal kidneys and ureter
B) nephritis caused by IBV
Egg qua lity

In laying flocks affected by IBV, a decline in egg production and quality occurs,
m addition to more or less evident respiratory symptoms
Normal eggs compared with typical altered

egg shells (ri ght and left eggs) alter
IB infection
Normal reproductive tract compared with

an atrophic oviduct of a hen of the same
age, affected by IB (bottom) and a
physiologically silent oviduct (top)
Infectious bursal disease (IBD)

< 3 weeks
3-6 weeks
AGE
Control for:
- presence of IBD in the area
- vaccination schedule and
type of vaccine
Control for:
- presence of IBD in the area
- vaccination schedule
and typ e o f vaccine
- sym ptom s: nearly absent,
atrophy of bursa of
Fabricius (BF)
Laboratory
Virus isolation
and histological
lesions
negative
coccidia, salm onella, c o li
Mortality
D iagnosis o f IBD
O ccu rred
in past weeks
Present at the
tim e of visit
positive
Symptoms:
depression, anorexia,
diarrhoea, ru ffle d feathers
Necropsy:
- lesions in th e BF
(edema and haemorrhages);
- proventriculus haemorrhages;
- m uscle haem orrhages
- C ontrol health status of

the g roup
- Size of BF or atrophy
~ Take b lo o d sam ples fo r
serology
NO
C heck and a d ju stm e n t of
vaccination schedule
C on trol of vaccine
a d m in istra tio n
Vaccine type and strain
D iagnosis of IBD
Verify verifica tio n and

a d ju stm e n t o f vaccination
schedule
Control of vacrine administration
Vaccine typ e and strain
90
YES
i kS
Further examinations
take b lo o d sam ples d u rin g
the visit and 2-3 w eeks later
Laboratory
S erological tests
NO
C ontrol fo r o th e r diseases
IBD Lesions
lesions m bursa of
F ab ricius an d in m uscles
Presence of edema in the bursa of Fabricius
Presence of haemorrhages in the bursa
Haemorrhages in muscle
Micrograph of a bursa of Fabricius in a normal chicken and in a chicken after infection

| with IBD virus
MAREK'S DISEASE
Marek's disease (MD)
NO
C ontrol fo r o th e r diseases,
p articula rly lym p h o id and m yelo id
leucosis
Clinic/Necropsy:
- leg paralysis
- depression
- n eoplastic lesions in viscera
and skin
- nerve lesions
YES
Feather tip exam ination by AGD test
D iagnostic and prog n ostic value

for M D
- Virus vaccinal type and strain

- C ontrol of vaccine a dm in istra tio n
in hatchery
92
MD Lesions
Nerve lesions
Characteristic leg paralysis in a 12 week

old chicken due to MD
Sciatic nerve lesion due to neoplastic

infiltration caused by MD
Vagal nerve lesion clue to neoplastic infiltration caused by MD, compared with a
normal nerve (at the top)
V isceral le s io n s
Neoplastic lesions as infiltrates or nodules in the liver due to MD
Neoplastic lesions in the proventriculus due ro MD
Skin lesions
Skin nodules in a bird affected by MD
Microscopic section of a lymphoma caused

by MD; the lesion 15 characterized by a
pleomorphic population of tumoral cell
that infiltrate organs
AGD test for diagnosis and prognosis of

MD; in affected birds, a precipitation line
is observed between the central positive
serum well and feather tips
93

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Poultry Disease Manual

Pro/. Antonio Zanella

INFECTIOUS BURSAL DISEASE

m PULLORUM DISEASE AN D FOWL TYPHOID

- MYCOPLASMA GALL1SEPTICUM INFECTION

FACTORS INFLUENCING VACCINATION

m anual o n in fectious diseases, m ostly o f intensively-reared dom estic

Immunity in birds: general concepts

organ s, the thym us an d the b u rsa o f

structures, which include bone-marrow,

spleen, H arderian gland, Peyers patches,

in the body (birds lack the equivalen t

o f m am m alian lymph nodes).

tiatio n of the germ in al im m u n e cells

in to two types o f ly m p h ocy tes takes

im m u n e cells, such as m onocytes and m acrop h ages, granulocytes, |

with the antigens which

In bird s, three m ain classes o f Ig have b een rep orted :

flu id s, as bile an d resp on sible fo r m u co sal im m unity; the existence

disease virus strain s N D V 6 / 1 0 o r H itch n er B l) ;

provided q u ite quickly after adm inistration (local im m unity system),

alth o u g h a n tib o d ie s a p p e a r in th e se ru m on ly a fte r 10-15 days.

o f age a n d it is su ffic ie n t to vaccin ate p a re n t m e n s befo re g o in g

T h e y are o fte n a sso c ia te d w ith a d ju v a n ts, w h ich stre n g th e n the

Clinical signs and lesions