TUGAS MATA KULIAH

FISIOLOGI DAN TEKNOLOGI REPRODUKSI VETERINER

(Anatomi Reproduksi Betina dan Jantan)

Reproduksi Jantan pada Kuda

Oleh
Gede Wiyasa Ardy Nugraha
1709511059
Kelas B

FAKULTAS KEDOKTERAN HEWAN

UNIVERSITAS UDAYANA
TAHUN 2020
 KATA PENGANTAR

Puji syukur saya panjatkan ke hadirat Tuhan Yang Maha Esa, karena dengan
rahmat dan hidayah-Nya saya dapat menyelesaiakan paper yang berjudul
“Reproduksi Jantan pada Kuda”.
Meskipun banyak rintangan dan hambatan yang saya alami dalam
proses pengerjaannya, tapi saya berhasil menyelesaikannya dengan baik. Adapun
paper ini dibuat guna memenuhi tugas mata kuliah Fisiologi dan Teknologi
Reproduksi Veteriner di Fakultas Kedokteran Hewan Universitas Udayana.
Saya sebagai penulis mengucapkan rasa berterimakasih yang sebesar-
besarnya kepada semua pihak yang telah membantu saya sehingga dapat
menyelesaikan paper ini tepat waktu. Saya yakin paper ini masih jauh dari nilai
kesempurnaan. Oleh karena itu, kritik dan saran yang bersifat membangun sangat
diharapkan oleh penulis demi menjadikan paper ini bisa lebih baik lagi.
Semoga paper "Reproduksi Jantan pada Kuda" memberikan informasi yang
berguna bagi masyarakat serta bermanfaat untuk pengembangan wawasan dan
peningkatan ilmu pengetahuan bagi kita semua.

Denpasar, 15 Maret 2020

Penulis

i
 DAFTAR ISI

Cover
Kata Pengantar .................................................................................................... i
Daftar Isi ............................................................................................................ ii
Daftar Gambar................................................................................................... iii
Daftar Lampiran ................................................................................................ iv
Bab I Pendahuluan
1.1 Latar Belakang ............................................................................................ 1
1.2 Rumusan Masalah ....................................................................................... 2
1.3 Tujuan Penulisan ......................................................................................... 2
Bab II Tinjauan Pustaka
2.1 Penis ............................................................................................................ 3
2.2 Kelenjar bulbourethral ................................................................................ 5
2.3 Kelenjar Prostat ........................................................................................... 5
2.4 Vesikula Seminalis...................................................................................... 5
2.5 Kelenjar Ampula ......................................................................................... 6
2.6 Vas Deferens ............................................................................................... 6
2.7 Epididimis ................................................................................................... 7
2.8 Testis ........................................................................................................... 7
Bab III Penutup
3.1 Simpulan ................................................................................................... 12
3.2 Saran.......................................................................................................... 12
Daftar Pustaka .................................................................................................. 13

ii
 DAFTAR GAMBAR

Gambar 2.1 Saluran reproduksi kuda jantan: representasi diagram ................... 3

Gambar 2.2 Saluran reproduksi kuda jantan setelah pemotongan dan
pembedahan, kelenjar aksesori tidak termasuk ................................................... 4
Gambar 2.3 Potongan melintang vas deferens kuda jantan ................................ 6
Gambar 2.4 Ilustrasi diagram penampang vertikal testis kuda jantan ................ 8
Gambar 2.5 Testis kuda yang dibedah menggambarkan jaringan testis,
epididimis, vas deferens dan pleksus pampiniformis .......................................... 9
Gambar 2.6 Jalur normal testis turun pada kuda jantan .................................... 10
Gambar 2.7 Bagian penampang melintang tubulus seminiferus dalam testis kuda
jantan, menggambarkan pembagian meiosis bertahap spermatogonia menjadi
spermatozoa ...................................................................................................... 11

iii
 DAFTAR LAMPIRAN

Lampiran 1. Reproductive Physiology Of The Stallion. Vi. Seminal And

Behavioral Characteristics.
Lampiran 2. Testicular Teratoma in a Unilateral Right-Sided Abdominal
Cryptorchid Horse.
Lampiran 3. Equine Testicular Lesions Related to Invasion by Nematodes.

iv
 BAB I PENDAHULUAN

1.1. Latar Belakang

Kuda telah didomestikasi lebih dari 6.000 tahun yang lalu di daerah stepa
yang sekarang dikenal dengan daerah Rusia Selatan dan Ukraina. Sejak itu
kuda mempunyai banyak manfaat yang berhubungan dengan manusia.
Penduduk asli Indonesia telah beternak kuda sebelum kedatangan Eropa. Kuda
hidup pada saat itu di alam bebas dan sangat tergantung pada kebaikan alam
sehingga kuda yang dipelihara memiliki kualitas rendah. Kedatangan Portugis
dan Belanda ke Indonesia memiliki andil memperbaiki ras kuda lokal,
termasuk memperbaiki cara beternak seperti cara pemberian makan yang baik,
perawatan kuda, serta petunujuk-petunjuk lain yang berhubungan dengan kuda
(Putriana, 2011).
Ternak kuda termasuk komoditas ternak yang ada di Indonesia dan belum
mendapat perhatian yang proporsional baik oleh pemerintah maupun oleh
masyarakat. Keberadaan ternak kuda dinilai cukup strategis karena fungsinya
sebagai ternak kerja (salah satunya adalah kuda penarik andong) dan memiliki
nilai estetika yang menarik. Penelitian tentang ternak kuda sampai saat ini
belum banyak dilakukan oleh pakar di bidang peternakan bahkan publikasi
ilmiah tentang ternak kuda di Indonesia sangat langka, pembahasan dan diskusi
mengenai perkembanganya hampir tidak mendapat perhatian (Setyobudi,
2009).
Dewasa ini, terjadi penurunan jumlah kelahiran kuda dikarenakan masih
kurangnya tempat pembudidayaan kuda yang memenuhi standar, serta masih
kurangnya pengetahuan para peternak kuda tentang siklus reproduksi kuda. Hal
tesebut menyebabkan para peternak kuda tidak dapt menentukan masa kawin
atau musim kawin yang tepat bagi kuda sehingga jumlah kelahiran kuda tidak
mencapai titik optimal.
Karena hal tersebut maka perlu dipelajari tentang reproduksi pada kuda,
selain sebagai tugas mata kuliah fisiologi dan teknologi reproduksi veteriner,
juga menjadi tambahan pengetahuan umum dalam memahami sistem
reproduksi kuda.

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1.2. Rumusan Masalah
Berdasarkan latar belakang yang telah dibuat dapat dibuat rumusan
masalah yaitu “Bagaimana anatomi dan fisiologi reproduksi dari kuda jantan?”

1.3. Tujuan Penulisan

Berdasarkan rumusan masalah diatas dapat dibuat tujuan penulisan yaitu
untuk mengetahu anatomi dan fisiologi reproduksi dari kuda jantan.

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 BAB II TINJAUAN PUSTAKA

2.1. Penis
Penis kuda jantan dapat dibagi menjadi glans penis, body atau shaft dan
roots. Dalam posisi istirahat, kuda terletak masuk dan dilindungi dalam
preputium sehingga tidak terlihat dan dipegang dalam posisi ini oleh otot.
Preputium adalah penutup berlipat ganda pada penis, yang melipat kembali
dengan sendirinya untuk memberikan perlindungan dua kali lipat. Di dalam
lipatan dalam terletak ujung penis, glans penis (atau rose), memberikan
perlindungan tambahan area sensitif ini. Dengan tinggi 5 mm, dari pusat glans
penis, terletak pintu keluar uretra. Di sekitar tonjolan ini terletak fossa uretra
dan, di bawahnya, divertikulum dorsal, keduanya sering diisi dengan smegma,
sekresi merah-coklat dari kelenjar prepubital yang melapisi preputium dan
epitel debris. Daerah-daerah ini juga menyediakan lingkungan yang ideal untuk
bakteri, sering mengandung bakteri penyakit kelamin (VD), seperti Klebsiella
pneumoniae, Taylorella equigenitalis dan Pseudomonas aeruginosa.

Gambar 2.1 Saluran reproduksi kuda jantan: representasi diagram.

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Gambar 2.2 Saluran reproduksi kuda jantan setelah pemotongan dan pembedahan,
kelenjar aksesori tidak termasuk.

Penis kuda jantan melekat pada bagian bawah tulang panggul oleh dua
akar. Di sini pada asalnya penis dipegang pada posisinya oleh ikatan ligamen
pada pelvis. Uretra, dari kandung kemih, terhubung dengan vas deferens dan
di antara kedua akar sebelum memasuki tubuh penis. Body penis kuda
mengandung persentase besar ereksi, tidak seperti fibro, jaringan dan karena
itu disebut hemodinamik (bereaksi terhadap peningkatan tekanan darah).
Tubuh utama penis dibagi menjadi dua bagian, yaitu corpus cavernosus
uretra bagian bawah dan penis corpus cavernosus bagian atas. Melalui corpus
cavernosus uretra melalui uretra, dikelilingi oleh beberapa trabekula (lembaran
jaringan ikat) yang menutupi area kecil dari jaringan ereksi, semua tertutup
dalam otot bulbospongiosus. Penis corpus cavernosus, yang merupakan bagian
terbesar dari penis, berisi jaringan padat trabekula, jaringan otot yang terkait
dan rongga yang tersebar, membentuk jaringan ereksi utama penis. Penis

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 corpus cavernosus terkandung dalam tunica albuginea, kapsul atau lembaran
fibroelastik, yang mempertahankan integritas penis tetapi masih
memungkinkan penggandaan ukuran yang terlihat saat ereksi. Akhirnya,
melalui di sepanjang bagian bawah penis adalah otot retractor, kontraksi yang
mengembalikan penis ke dalam preputium. Jaringan ereksi utama glans penis
adalah kelanjutan dari corpus cavernosus urethra, corona glandis. Jumlah besar
jaringan ereksi ini, yang tidak dibatasi oleh kapsul fibrosis penis corpus
cavernosus, memungkinkan perluasan yang lebih besar (hingga tiga kali) dari
area ini saat ejakulasi.

2.2. Kelenjar bulbourethral

Kelenjar bulbourethral, atau kelenjar Cowper, adalah kelenjar aksesori
yang terletak paling dekat dengan akar penis. Mereka berpasangan dan
berbentuk oval, sekitar 2 cm dan terletak di kedua sisi uretra. Sekresi mereka
jelas, encer dan berair dan membentuk bagian dari fraksi kaya sperma utama.
Mereka juga berkontribusi pada fraksi pra-sperma. Sebagai bagian dari fraksi
pra-sperma, mereka membantu membersihkan urin dan bakteri yang
dikumpulkan dalam uretra sebelum ejakulasi. Sekresi mereka juga dapat
bertindak sebagai pelumas, memudahkan perjalanan sperma.

2.3. Kelenjar Prostat

Kelenjar prostat adalah struktur bilobed dengan lubang keluar tunggal ke
uretra, terletak di antara kelenjar bulbourethral dan ampula. Sekresi prostat
pada kuda jantan bersifat basa dan tinggi protein, asam sitrat dan zinc.
Signifikansi ini tidak jelas. Sekresi kelenjar prostat memberikan kontribusi
yang signifikan terhadap fraksi pra-sperma.

2.4. Vesikula Seminalis

Kelenjar vesikular, atau vesikula seminalis, berpasangan dalam struktur
dan terletak di kedua sisi kandung kemih, panjangnya sekitar 16-20 cm.
Mereka lobed dan dapat dibandingkan dengan kenari besar dalam penampilan
luar. Mereka mengeluarkan sejumlah besar plasma mani, dengan konsentrasi

5
 tinggi kalium, asam sitrat dan gel. Sekresi mereka membentuk bagian dari
fraksi pasca-sperma yang kaya sperma dan seperti gel. Fungsi mereka dan
karenanya volume sekresi tergantung pada konsentrasi testosteron yang
beredar. Dengan demikian, kontribusi mereka terhadap plasma mani menurun
secara signifikan selama musim tidak kawin.

2.5. Kelenjar Ampula

Kelenjar ampula terletak berpasangan keluar lipatan dari vas deferens
dimana bertemu uretra. Sekresi kelenjar ampula mengandung ergothionine
konsentrasi tinggi, agen antioksidan, melindungi bahan kimia yang rentan
dalam semen dari oksidasi. Sekresi ampula sebagian besar terkait dengan fraksi
pra-sperma.

2.6. Vas Deferens

Vas deferens menghubungkan epididimis ke uretra sebelum melewati
kelenjar aksesori dan masuk ke penis. Ia memiliki diameter 0,5-1 cm, dengan
dinding berotot tebal, terdiri dari tiga lapisan otot; oblique bagian dalam,
lingkaran tengah dan longitudinal luar. Lapisan otot ini secara aktif mendorong
sperma dan cairan di sekitarnya dari testis ke penis. Lumen duktus kecil dan
dilapisi lipatan, terutama di dekat epididimis, memaksimalkan luas permukaan
sehingga membantu penyimpanan sperma dan reabsorpsi cairan testis.

Gambar 2.3 Potongan melintang vas deferens kuda jantan.

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 Vas deferens, suplai saraf testis, pembuluh darah arteri dan vena dan otot
kremaster keluar dari rongga tubuh melalui kanalis inguinalis. Otot cremaster,
yang dibagi menjadi beberapa bagian internal dan eksternal, bertanggung
jawab untuk menarik testis ke arah tubuh sebagai respons terhadap dingin,
ketakutan, dll.

2.7. Epididimis
Epididimis pada kuda jantan terletak di atas testis. Ini terdiri dari tubulus
berbelit-belit panjang dan dibagi menjadi tiga bagian: kepala (caput); tubuh
(corpus); dan ekor (cauda). Kepala epididimis dihubungkan oleh beberapa
saluran ke rete testis, yang terus menuju ujung ekor, saluran ini bergabung dan
membentuk satu saluran yaitu vas deferens. Lapisan tubulus ini sangat terlipat,
sangat mirip dengan ujung epididimis vas deferens. Lipatan ini memiliki
mikrovili tambahan, meningkatkan luas permukaan lebih jauh dan
memfasilitasi reabsorpsi sekresi testis untuk memusatkan sperma dan
meningkatkan kapasitas penyimpanannya. Sangat penting bahwa semua
sperma menghabiskan periode waktu, hingga 7 hari, dalam epididimis untuk
menjadi dewasa. Pematangan tersebut sangat penting sehingga sperma yang
dilepaskan mampu berkembang lebih lanjut (kapasitasi) dalam saluran wanita,
memungkinkan mereka untuk membuahi sel telur yang menunggu. Jika mereka
tidak diteruskan ke vas deferens sebagai akibat dari ejakulasi, mereka
mengalami degenerasi dan diserap kembali, memungkinkan pasokan sperma
segar terus-menerus untuk mempertahankan tingkat pembuahan. Ujung ekor
vas deferens, oleh karena itu, bertindak sebagai tempat penyimpanan sperma
dan juga berkontribusi terhadap plasma mani dengan mengeluarkan
glycerylphosphorylcholine (GPC).

2.8. Testis
Testis bersifat gametogenik (tempat produksi sperma) dan memiliki
fungsi endokrin. Testis menggantung di luar tubuh kuda jantan untuk menjaga
suhu sekitar 3 ° C di bawah suhu tubuh (mis. 35-36 ° C daripada 39 ° C).
Produksi sperma dimaksimalkan pada suhu yang lebih rendah ini. Peningkatan

7
suhu testis karena penyakit atau peradangan skrotum, testis atau epididimis
spermatogenesis mengalami penurunan yang signifikan. Ini sementara tetapi
durasi disfungsi testis terkait dengan durasi kenaikan suhu. Suhu testis biasanya
dikontrol melalui otot-otot kremaster, yang dapat menarik testis ke arah tubuh,
bersama dengan banyak kelenjar keringat pada skrotum dan mekanisme
pertukaran panas kontra-arus arteriovenosa yang disediakan oleh pleksus
pampiniformis. Pleksus pampiniformis memungkinkan arteri testis membelah
menjadi jaringan kapiler yang padat sebelum memasuki testis. Dengan
demikian, berhubungan erat dengan pasokan vena yang kembali, juga dibagi
menjadi jaringan kapiler. Darah memasuki testis, oleh karena itu testis
kehilangan panas ketika aliran balik ke vena. Pengaturan semacam itu
memastikan bahwa arteri testis menjadi dingin sebelum masuk ke dalam testis
dan vena testis dihangatkan sebelum masuk kembali ke tubuh.

Gambar 2.4 Ilustrasi diagram penampang vertikal testis kuda jantan.

8
Gambar 2.5 Testis kuda yang dibedah menggambarkan jaringan testis, epididimis,
vas deferens dan pleksus pampiniformis.

Testis terletak di dalam selubung kulit, disebut skrotum, di bawahnya

terdapat tunica vaginalis, yang kontinu dengan lapisan peritoneum rongga
tubuh naik melalui kanal inguinal. Pada janin, testis turun dari posisi dekat
ginjal, melalui kanalis inguinalis dan ke skrotum segera setelah lahir.
Kegagalan satu atau kedua testis untuk turun sepenuhnya mungkin tidak berarti
kuda jantan tidak subur, karena testis di dalam rongga tubuh masih dapat
menghasilkan sperma sehingga jumlah sperma, bagaimanapun, biasanya jauh
berkurang. Kuda jantan dengan hanya satu testis turun disebut cryptorchid atau
rig. Kondisi ini selanjutnya dapat didefinisikan sebagai unilateral, bilateral,
inguinal atau abdominal, tergantung pada apakah satu atau kedua testis telah
gagal turun dan seberapa jauh penurunan telah berkembang.

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 Gambar 2.6 Jalur normal testis turun pada kuda jantan

Testis kuda biasanya terletak dengan sumbu panjang horizontal kecuali

ditarik ke arah tubuh, ketika mereka mungkin sedikit berubah. Sumbu panjang
biasanya 6-12 cm dengan tinggi dan lebar masing-masing 4-7 cm dan 5-6 cm.
Testis dapat berbobot 300-350 g per pasang. Ukuran mereka meningkat secara
allometrik dengan pertumbuhan tubuh secara umum sampai ukuran tubuh akhir
telah tercapai, pada sekitar usia 5 tahun.
Di bawah tunica vaginalis, kapsul fibrosa, tunica albuginea, mengelilingi
setiap testis yang terpisah. Lembaran jaringan fibrosa ini memasuki tubuh testis
dan membaginya menjadi lobus. Setiap lobus adalah massa tubulus seminiferus
berbelit-belit dengan area antar tubulus. Setiap area sebagian besar
bertanggung jawab atas satu dari dua fungsi yaitu sitogenik (tubulus
seminiferus dan sel Sertoli) atau endokrin (jaringan antar tubular dan sel
Leydig). Sel-sel Sertoli yang melapisi tubulus seminiferus bertindak sebagai
sel perawat, memberi nutrisi dan membantu spermatozoa berkembang di lumen
tubulus. Selain itu, mereka juga fagosit, mencerna sel germinal yang
memburuk dan sisa tubuh. Mereka mengeluarkan cairan luminal dan protein,
dan juga membentuk penghalang testis-darah, memberikan perlindungan bagi
sperma dari penolakan imunologis dan komunikasi sel-ke-sel. Jumlah sel
Sertoli bervariasi sesuai musim, secara signifikan lebih besar selama musim
kawin, ketika mereka secara aktif terlibat dalam produksi sperma. Dengan

10
 demikian, sel Sertoli memiliki kontrol dominan terhadap produksi sperma.
Peningkatan jumlah sel Sertoli ini disertai dengan peningkatan panjang tubulus
seminiferus yang sesuai.

Gambar 2.7 Bagian penampang melintang tubulus seminiferus dalam testis kuda
jantan, menggambarkan pembagian meiosis bertahap spermatogonia menjadi
spermatozoa.

Sel-sel Leydig, yang ditemukan di ruang antar tubulus di sekitar tubulus

seminiferus, mengeluarkan hormon yang sebagian besar bertanggung jawab
untuk produksi sperma dan pengembangan karakteristik dan perilaku tubuh
pria secara umum.

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 BAB III PENUTUP

3.1. Simpulan
Penis kuda jantan dapat dibagi menjadi glans penis, body atau shaft dan
roots. Tubuh utama penis dibagi menjadi dua bagian, yaitu corpus cavernosus
uretra bagian bawah dan penis corpus cavernosus bagian atas. Kelenjar
bulbourethral, atau kelenjar Cowper, adalah kelenjar aksesori yang terletak
paling dekat dengan akar penis. Mereka berpasangan dan berbentuk oval,
sekitar 2 cm dan terletak di kedua sisi uretra. Kelenjar prostat adalah struktur
bilobed dengan lubang keluar tunggal ke uretra, terletak di antara kelenjar
bulbourethral dan ampula. Kelenjar vesikular, atau vesikula seminalis,
berpasangan dalam struktur dan terletak di kedua sisi kandung kemih,
panjangnya sekitar 16-20 cm. Kelenjar ampula terletak berpasangan keluar
lipatan dari vas deferens dimana bertemu uretra. Vas deferens menghubungkan
epididimis ke uretra sebelum melewati kelenjar aksesori dan masuk ke penis.
Ia memiliki diameter 0,5-1 cm, dengan dinding berotot tebal, terdiri dari tiga
lapisan otot; oblique bagian dalam, lingkaran tengah dan longitudinal luar.
Testis bersifat gametogenik (tempat produksi sperma) dan memiliki fungsi
endokrin. Testis menggantung di luar tubuh kuda jantan. Testis terletak di
dalam selubung kulit, disebut skrotum, di bawahnya terdapat tunica vaginalis,
yang kontinu dengan lapisan peritoneum rongga tubuh naik melalui kanal
inguinal. Di bawah tunica vaginalis, kapsul fibrosa, tunica albuginea,
mengelilingi setiap testis yang terpisah. Lembaran jaringan fibrosa ini
memasuki tubuh testis dan membaginya menjadi lobus. Setiap lobus adalah
massa tubulus seminiferus berbelit-belit dengan area antar tubulus. Sel-sel
Leydig, yang ditemukan di ruang antar tubulus di sekitar tubulus seminiferus.

3.2. Saran
Saran dari penulis untuk penulisan ini masih jauh dari kata sempurna,
masih banyak sumber-sumber yang lebih compatible mengenai materi ini.
dimohonkan para pembaca tidak hanya menggunakan paper ini sebagai acuan
tetapi bisa mencari di tempat lain dengan informasi yang lebih lengkap.

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 DAFTAR PUSTAKA

M.C.G. Davies Morel, 2003, Equine Reproductive Physiology, Breeding and Stud
Management 2nd Edition, CABI Publishing, Institute of Rural Studies
University of Wales, Aberystwyt, UK

B. W. Pickett, L. C. Faulkner, G. E. Seidel, Jr., W. E. Berndtson and J. L. Voss,

1976, Reproductive Physiology Of The Stallion. Vi. Seminal And
Behavioral Characteristics, Journal Of Animal Science, Vol. 43, No. 3, hh
617-625.

Ugolini, W, Santos, F, Costa, G, Oliveira, H, Folchini, N, Machado, T, Zannella, R

& Alves, L 2019, Testicular Teratoma in a Unilateral Right-Sided
Abdominal Cryptorchid Horse, Acta Scientiae Veterinariae, Vol. 47(Suppl
1): 409, hh 1-5.

Marino, G, Zanghi, A, Quartuccioo, M, Cristarella, S, Giuseppe, M, & Catone, G

2009, Equine Testicular Lesions Related to Invasion by Nematodes, Journal
of Equine Veterinary Science, Vol 29, No 10, hh 728-733.

13
 REPRODUCTIVE PHYSIOLOGY OF THE STALLION.
VI. SEMINAL AND BEHAVIORAL CHARACTERISTICS 1

B. W. Pickett, L. C. Faulkner, G. E. Seidel, Jr., W. E. Berndtson

and J. L. Voss 2

Colorado State University, F o r t Collins 8 0 5 2 3

SUMMARY permit prediction- of reproductive character-

First and second ejaculates were collected
from five mature Quarter Horse stallions each
Tuesday for 13 months. Means and statistical
comparisons of the seminal characteristics for
260 first and second ejaculates were: gel-free
semen (ml) 58.4, 49.6 (P<.05); gel (ml) 7.1, 2.0
(P>.05); total seminal volume (ml) 65.5, 51.6
(P<.05); spermatozoa/ml (106) 281, 170
(P<. 01); spermatozoa/ejaculate ( 109 ) 14.7, 7.9
(P<.01); motility (%) 73, 74 (P>.05); pH 7.47,
7.59 (P<.01) and osmolality of seminal plasma
(m0s) 302, 300 (P>.05), respectively. Behav-
ioral characteristics of stallions associated with
collection of first and second ejaculates were:
reaction time (sec) 211, 231 (P>.05) and
mounts per ejaculate 1.8, 1.7 (P>.05). Signifi-
cant (P<.01) stallion effects were found for all
first and second ejaculate criteria except motil-
ity in first ejaculates and reaction time required
for second ejaculates (P<.05). Seasonal effects
(P<.01) for first ejaculates were observed for
seminal volumes, spermatozoa/ejaculate, pH,
reaction time and mounts per ejaculate. Sea-
sonal effects on second ejaculate characteristics
were significant (P<.01) for gel volume (ml),
spermatozoa/ml (106) and spermatozoa/ejacu-
late (109), pH and mounts per ejaculate.
Although a number of significant correlations
among the reproductive characteristics studied
and plasma testosterone levels were obtained,
the coefficients of correlation were too small to
i Animal Reproduction Laboratory, Department of
Physiology and Biophysics. This investigation was
supported in part by grants-in-aid from American
Breeders Service, The Grayson Foundation and the
National Association of Animal Breeders. Portions of
these data were presented at the "First International
Symposium Upon Equine Reproduction," Cambridge,
England, 1974, and appeared in J. Reprod. Fertil.
(Suppl.) 23:25. 1975.
2Department of Clinical Sciences.
617
JOURNAL OF ANIMAL SCIENCE, Vol. 43, No. 3 (1976)
istics from individual testosterone values. Corre-
lations based on the pooled monthly means for
each stallion or calculated on a within-stallion
basis were similar in magnitude to simple
correlations.
The relationship among these characteristics
is discussed in relation to potential fertility
evaluation of stallions.
(Key Words: Stallion, Semen, Behavior, Sper-
matozoa, First and Second Ejaculates.)
INTRODUCTION
Potential fertility evaluation of stallions has
been neglected due to failure to identify the
factors responsible for the extreme variation in
seminal characteristics and the lack of informa-
tion on the relationship among these factors
and fertility. Season influences many of the
physical and chemical characteristics of stallion
semen as well as blood hormone levels, sexual
behavior and fertility of both sexes (Mann et
al., 1956; Nishikawa, 1959; Osborne, 1966;
Skinner and Bowen, 1968; Loy, 1970; Pickett
et al., 1970; Berndtson et al., 1974; Ginther,
1974; Gebauer et al., 1976). Other factors
affecting seminal characteristics are sexual stim-
ulation, frequency o f ejaculation, age, testicular
size and method of seminal collection (Nishika-
wa, 1959; Bielanski, 1964; Pickett et al., 1970;
1974b; 1975b,c; Pickett and Back, 1973;
Gebauer et al., 1974a,b,c; Swierstra et al.,
1974).
The purposes of this investigation were to
determine: 1) influence of stallion behavior on
seminal characteristics, 2) relationship of semi-
nal characteristics of first and second ejaculates
collected throughout 13 months to permit
seminal evaluation, irrespective of season, and
3) relationship between plasma testosterone
levels and seminal and behavioral character-
istics,

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618 PICKETT ET AL.
M A T E R I A L S A N D METHODS
First and second ejaculates of semen were
collected from each of five Quarter Horse
stallions ranging in age from 5 to 12 years each
Tuesday from April 1969 through May 1970.
Second ejaculates were collected one hour after
the first, or as soon thereafter as possible. The
artificial vagina (A.V.) used for seminal collec-
tion, techniques for preparation of the stallion,
methods for measuring spermatozoan motility
and concentration and pH of semen and gel
have been described (Pickett and Back, 1973).
Osmolality was determined with a Fiske os-
mometer (Model G-66x) on 2.0 ml of seminal
plasma obtained by centrifuging 3.0 ml of
semen at 10,000 g for 3 minutes.
The influence of stallion and month on
sexual behavior was studied by measuring reac-
tion time and number of mounts per ejacula-
tion. Reaction time was the total time (see) the
stallion was in visual contact with the mare
until the beginning of copulation.
Of the 560 planned seminal collections, one
first and four second ejaculates were not
obtained. The five failures were ignored in the
statistical analyses. Data from the fifth Tuesday
in any month were eliminated to equalize
numbers.
The data for the seminal and behavioral
characteristics were analyzed by the methods of
Snedecor and Cochran (1967). The data were
subjected to factorial analyses of variance with
two fixed factors, months and first and second
ejaculates, and one random factor, stallions.
The degrees of freedom for months were
partitioned into linear, quadratic and cubic
effects. A n y seasonal trends should be reflected
in significant quadratic and/or cubic effects.
Correlations between plasma testosterone
levels (Berndtson et al., 1974) and the other
reproductive characteristics were computed af-
ter pairing the testosterone concentration of
individual plasma samples with the value for
each reproductive characteristic for the seminal
collection of the previous day. Coefficients of
correlations were also determined after pooling
individual values within stallions on a monthly
basis.
RESULTS A N D DISCUSSION
Presented in figures 1 and 2 are the patterns
of seminal volume, motility, spermatozoal out-
put, pH and seminal plasma osmolality of the
first and second ejaculates for 13 months.
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Patterns of sexual behavior, measured by
mounts per ejaculate and reaction time, are
presented in figure 3. Highly significant (P<.01)
stallion effects were observed for all criteria in
first and second ejaculates except that motility
of spermatozoa in first ejaculates and reaction
time for collection of second ejaculates were
significant at the 5% level of probability.
Significant (P<.01) seasonal effects for first
ejaculates were observed for seminal volumes,
spermatozoa per ejaculate, pH, reaction time
and mounts per ejaculate. Seasonal effects on
second ejaculate characteristics were significant
(P<.01) for gel volume, spermatozoa per ml
and per ejaculate, pH and mounts per ejaculate.
The mean volume of gel-free semen in first
ejaculates was 58.4 compared to 49.6 ml in
second ejaculates (P<.05). Volume of first
ejaculates ranged from 44.8 to 80.8 ml in
March and July, respectively; second ejaculates
varied from 41.5 ml in April to 62.8 ml in June.
Mean volume of gel was 7.1 ml for all first
and 2.0 ml for all second ejaculates (P>.05).
However, in the 75 first ejaculates that con-
tained gel (29%), the mean gel volume was 27
ml, while 29 second ejaculates (11%) contained
an average of 19 milliliters. It appeared that the
seminal vesicles, which secrete the gelatinous
fraction, were slower to replenish their secre-
tion than the other secondary sex glands. This
may be due to an inability of certain stallions
to become sexually stimulated during certain
seasons or after an extended regimen of seminal
collection. Although no evidence was pre-
sented, Nishikawa (1959) reported that gel
served as a bacteriostat. Regardless of the
function of gel, semen from some stallions
rarely, if ever, contains gel, and yet, their
fertility appears normal. Thus, the presence Or
absence of gel in an ejaculate may not reflect
the fertility of that ejaculate. Further, when
mares are bred utilizing artificial insemination
(A.I.), the gel is a nuisance and is removed
during seminal collection (Pickett and Back,
1973). Fertility of semen appeared to be
normal (Pickett et al., 1974a), although the gel
was never mixed with the "sperm-rich" (Kosi-
niak, 1975) fraction.
The gel must be removed during seminal
collection to obtain a representative sample for
estimation of spermatozoal concentration. The
gel can be removed after seminal collection, but
more spermatozoa are lost (Pickett et al.,
1974b). This is an important consideration
when the maximum number of spermatozoa are
 STALLION SEMINAL CHARACTERISTICS 619

Figure 1. Monthly means of seminal characteristics of first and second ejaculates from April 1969 through
May 1970 (N = 260). Shaded areas indicate the breeding season.

needed for A.I. istic is probably of least importance, since

The mean total seminal volume was 65.5 and
51.6 ml (P<.05) for first and second ejaculates,
respectively. The range for first ejaculates was
45.4 ml in January to 103.8 in June; the range
for second ejaculates was 41.5 ml in April to
67.8 ml in June. The total volume of first and
second ejaculates increased 38 and 21%, respec-
tively, from winter to spring. Total seminal
volume, generally including gel, is the most
commonly reported seminal characteristic
(Pickett, 1968, 1970). However, this character-
excellent fertility can be obtained by A.I. with
1.5 ml of raw semen, and as little as .6 ml may
be adequate (Pickett et al., 1974a, 1975a). In
addition, up to 100 ml of semen can be
obtained without the occurrence of complete
ejaculation (Pickett and Voss, 1975). Further,
when stallions were used as teasers, gel volume
increased from an average of 8 to 23 ml, and
mean gel-free seminal volume increased from 37
to 63 mt without an increase in spermatozoal
output (Pickett and Voss, 1972).

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620 PICKETT ET AL.
Figure 2. Monthly means of seminal pH and
seminal plasma osmolality of first and second ejacu-
lates from April 1969 through May 1970 (N = 260).
Shaded areas indicate the breeding season. The X
indicates the mean pH for May, 1970.
The relationship between seminal volumes in
first and second ejaculates in this study is
reasonably consistent with other investigations
(Pickett et al., 1970, 1975c). However, low first
ejaculate volume followed by a high second
ejaculate volume is not u n c o m m o n when semen
is collected from inexperienced 2- or 3-year-old
stallions. The first may serve as a stimulus for
the second ejaculate.
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Figure 3. Monthly means of reaction time and
number of mounts required to collect first and second
ejaculates from April 1969 through May 1970 (N =
260). Shaded areas indicate the breeding season.
Stallion spermatozoa tend to clump or agglu-
tinate (Skinner and Bowen, 1968; Pickett et al.,
1970, 1974a). To reduce the influence of
clumping on motility, semen was extended in a
Tris-egg yolk diluent that prevented clumping
(Pickett et aL, 1974a). Mean motility of sper-
matozoa from first ejaculates was 73%, with a
range of 70 to 75%; mean motility in second
ejaculates was 74% and ranged from 70 to 77%
(P>.05). No influence of season on motility
was observed for first or second ejaculates.
Motility of spermatozoa from first ejaculates is
occasionally lower than motility of spermato-
zoa in second ejaculates. This difference gener-
ally disappears when the frequency of ejacula-
tion is sufficient to prevent excessive spermato-
zoal accumulation (Pickett and Voss, 1975).
 STALLION SEMINAL CHARACTERISTICS
This may mean that differences i n spermato-
zoan motility between first and second ejacu-
lates could be due to spermatozoal accumula-
tion in the excurrent ducts. However, when
semen was collected from normal stallions,
there was no difference in motility between
first and second ejaculates when ejaculates were
collected weekly. Moreover, frequency of col-
lection (from 1 to 6 per week) did not affect
the motility of spermatozoa (Pickett et al.,
1970, 1975c).
Since no differences were observed in sper-
matozoan motility (figure 1) and in only a few
of the chemical constituents of spermatozoa
(Gebauer et al., 1976) due to season, it is
tempting to suggest that any seasonal variation
in stallion fertility might be due to the female
or to a reduced daily spermatozoal o u t p u t
(DSO) and/or libido rather than a decrease in
fertility of the individual spermatozoon.
First ejaculates contained 281 x 106 sper-
matozoa per ml compared to 170 • 106 per ml
in second ejaculates (P<.01). Mean monthly
concentration of spermatozoa in first ejaculates
ranged from 224 to 321 x 106/mi in December
and August, respectively; second ejaculates
ranged from 111 in December to 234 • 106/mt
in May 1969 (figure 1). First ejaculates con-
tained a mean of 14.7 • 109 spermatozoa;
second ejaculates contained 7.9 x 109 (P<.01).
First ejaculates ranged from 10.2 • 109 in
January to 22.1 x 109 in July. Second ejacu-
lates ranged from 5.0 to 11.8 • 109 in
December and July, respectively. The differ-
ence in mean total spermatozoa per ejaculate
between first and second ejaculates was 53.7%,
and the difference in monthly means between
the ejaculates ranged from 46.9 to 56.9%.
In a previous 12-month study (Pickett et al.,
1970), the difference between the means of the
170-paired first and second ejaculates was
4 9 . 8 % . When the ejaculation frequency was
doubled to two ejaculates twice per week, the
percentage of spermatozoa in second ejaculates
was 45.8% of the total in first ejaculates
(Pickett et al., 1975c). Bielanski (1964) re-
ported that. second ejaculates contained 60.8%
as many spermatozoa as in first ejaculates. The
relationship of spermatozoal numbers in first
and second ejaculates was so consistent that the
second ejaculate could be used to determine
how representative the first ejaculate was o f a
normal ejaculation. If a stallion has been
sexually rested for at least four days and the
second ejaculate does not contain approximate-
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621
ly 50% of the total number of spermatozoa in
the first ejaculate, one of the following must be
suspected: 1) one of the ejaculates was incom-
plete, 2) the stallion's extragonadal spermato-
zoal reserves (EGR) had been depleted, 3)
excessive spermatozoa had accumulated in the
excurrent ducts (Pickett and Voss, 1975), 4)
the excurrent ducts are incapable of storing
large quantities of spermatozoa, or 5) the
stallion was sexually immature. This approxi-
mately 50% relationship (5.9 vs 11.7 x 109)
was also observed when one ejaculate per day
was collected at intervals of six times per week
compared to three times per week after the
EGR had been stabilized (Pickett et al., 1975c).
Although Nishikawa (1959) and Skinner and
Bowen (1968) found no difference in spermato-
zoal output due to season, Pickett et al. (1970)
reported a 52.0 and 46.2% reduction in sper-
matozoal output from the highest to the lowest
month in first and second ejaculates, respective-
ly. In this study, corresponding reductions
averaged 46.2 and 42.4% (figure 1). This
dramatic decrease in spermatozoal output prob-
ably represents a true decrease in spermato-
genesis. Since season influenced (P<.01) sper-
matozoal numbers for both ejaculates, season
must be considered when stallions are evaluated
for breeding soundness.
Daily spermatozoal production (DSP) in the
stallion was 8.0 x 109 and daily spermatozoal
o u t p u t (DSO) accounted for 87% of DSP
(Gebauer et al., 1974a). Since DSP was 8.0 •
109 spermatozoa and mean extragonadal transit
time was 7.5 to 11.0 days, EGR should range
from 60 to 88 x 109. The EGR of 11 stallions
24 hr after the last ejaculation was 58.6 x 109
(Gebauer et al., 1974b). When one ejaculate of
semen was collected from each of nine stallions
at frequencies of 1, 3 and 6 times/week, mean
numbers of spermatozoa per ejaculate were
13.5, 12.7 and 7.2 • 109, respectively (Pickett
et al., 1974c). Therefore, if the number of
spermatozoa per ejaculate is more than 2 to 3
days DSP, it is suspected that the stallion is
accumulating spermatozoa in the excurrent
ducts.
When time is critical, two ejaculates col-
lected one hour apart are minimal for a seminal
evaluation. Ideally, DSP should be determined,
which can be predicted from DSO (Gebauer et
al., 1974c). To obtain DSO, semen must be
collected daily for approximately 1 week, and
DSO is dependent upon events initiated approx-
imately two months prior to seminal collection
622 PICKETT ET AL.
(Swierstra et al., 1974).
The number of spermatozoa necessary for
maximum reproductive efficiency in horses has
not been established. No difference in fertility
was found between 100 and 500 • 106 motile
spermatozoa when mares were inseminated
every other day of estrus beginning on day 2 or
3 (Demick et al., 1976).
Testicular size determined by scrotal circum-
ference permits relatively accurate estimation
of spermatozoal output in bulls (Hahn et al.,
1969). Stallion testicular weight was signifi-
cantly correlated with DSP and DSO (.77), as
expected since 21 • 106 spermatozoa were
produced per gram of testicular parenchyma
(Gebauer et al., 1974b,c). However, in vivo
testicular measurements accounted for only
32% of the variation in DSO (Gebauer et al.,
1974c). In spite of the relatively poor correla-
tions, the testes should be palpated and mea-
sured during a routine breeding soundness
examination, because a stallion with small
testes is a potentially poor producer of sper-
matozoa. The number of mares that can be
bred utilizing A.I. or natural service, to a lesser
extent, is dependent upon the number of
spermatozoa available for ejaculation.
Illustrated in figure 2 are the monthly
changes in pH. The mean pH of gel-free semen
from first ejaculates was 7.47 compared to 7.59
for second ejaculates (P<.01). The range for
first ejaculates was 7.36 to 7.57 (P<.01) while
the range for second ejaculates was 7.46 to 7.70
(P(.01). The pH is a useful diagnostic tool for
seminal evaluations, because in a "normal situa-
tion" the pH of second ejaculates was always
higher than corresponding first ejaculates (Pick-
ett et al., 1970, 1975c), regardless of season.
When the pH of a second ejaculate is lower
than the corresponding first ejaculate, the first
ejaculate must be considered incomplete or
abnormal. The pH of accessory sex gland fluid,
devoid of epididymal contributions will be
approximately 8.0. Kirton et al. ( 1 9 6 4 ) c o l -
lected four consecutive ejaculates of semen
from bulls within 1 hr and found that pH
increased from 6.26 to 6.92. These investigators
postulated that the contributions of the seminal
vesicles to the ejaculate was relatively constant,
but the contributions of the epididymides
decreased with repetitive ejaculation. This is an
extremely important observation in relation to
potential fertility evaluation of stallions, since
it is possible to obtain relatively large quantities
of seminal fluid and some spermatozoa without
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the occurrence of ejaculation, or at least with-
out a contribution from the epididymides
(Pickett and Voss, 1975). This postulation is in
excellent agreement with Seidel and Foote
(1970) who calculated that the bull epididy-
mides contributed 31 to 36% to the seminal
volume of first ejaculates compared to 19% to
second ejaculate volume. Therefore, when pH
of a first ejaculate is higher or equal to the pH
from a corresponding second ejaculate, particu-
larly if the normal relationship in spermatozoal
output is altered, the first ejaculate must be
considered incomplete or abnormal.
We believe the percentage contribution to
volume is higher from the accessory sex glands
of the stallion and less from the epididymides
in second ejaculates than in the bull, primarily
because the volume of the second ejaculate
does not change, in relation to first ejaculates,
as dramatically as spermatozoal output. Second
ejaculates from bulls usually contain 56.2 to
91.6% of the number of spermatozoa found in
first ejaculates (Kirton et al., 1964; MacMillan
et al., 1967; Pickett and Komarek, 1967). The
stallion ejaculate, after three days sexual rest,
normally contains more total spermatozoa than
the bull ejaculate. Further, the ductuli defer-
entia and ampullae contain sufficient spermato-
zoa for a complete ejaculate (Gebauer et al.,
1974b), whereas the corresponding structures
in the bull contain too few spermatozoa for a
complete ejaculate (Bialy and Smith, 1958).
The seminal vesicles contribute significantly
to the bovine ejaculate, which must correspond
to the bulbo-urethral glands in the stallion,
because the seminal vesicles in the stallion
secrete gel (Mann, 1964). The gel has a higher
pH than gel-free semen (figure 2), but probably
contributes very little to the seminal pH since it
is removed during collection. For pH to be of
diagostic value, the gel must be separated from
the "sperm-rich" fraction because: 1) the pH of
gel-free semen in the first ejaculate is lower that
that of second ejaculates, 2) pH of gel is higher
than that of gel-free semen, 3) there is a greater
quantity of gel secreted in first than in second
ejaculates, and 4) if the gel and "sperm-rich"
fractions mix during collection, the difference
in seminal pH beteen first and second ejaculates
would be greatly reduced.
Mean osmolality of seminal plasma from
first and second ejaculates was 302 and 300
m0s, respectively. There was no difference
between ejaculates or differences due to season
for either ejaculate. The range by month was
 STALLIOoNSEMINAL CHARACTERISTICS
297 to 312 m0s for first and 297 to 305 m0s
for second ejaculates.
Although most of the variation in osmolality
was due to stallion (P<.01), for both ejaculates,
the value of this measuremefit in potential
fertility evaluation of stallions is unknown. For
storage of stallion semen, either liquid or
frozen, the diluent must be compatible with the
spermatozoa. Unfortunately, the optimal osmo-
lality for a stallion seminal extender has not
been established. Pickett et al. (1974a) have
shown that a cream-gel extender, with an
osmolality of 347.6 m0s will provide fertility
equal to raw semen if used immediately after
dilution. However, after storage for 24 hr there
was a significant decrease in fertility (Demick et
al., 1976). Rajamannan et al. (1968) reported a
range of 290 to 310 m0s for stallion seminal
plasma, but found that an extender with an
osmolality of 330 provided superior survival of
equine spermatozoa after thawing.
Even though seminal quality may be excel-
lent, if libido is poor, fertility may be seriously
reduced. Thus, a thorough knowledge of sexual
behavior is essential for treating impotence and
other forms of abnormal sexual behavior (Pick-
ett and Voss, 1975). Mean reaction time was
211 sec for first ejaculates compared to 231 for
second ejaculates (P>.05). First-ejaculation re-
action time ranged from 47 sec in May 1969 to
721 in December; the range for second ejacu-
lates was 40 to 957 sec in May 1969, and
January 1970, respectively. Because of ex-
treme variation, differences due to season were
significant (P<.01) for only first ejaculates. The
fact that semen was collected from these
stallions twice per week, starting in May, might
have caused an increase in reaction time in the
fall and winter. However, as the breeding season
approached, reaction time decreased, which
appeared to be related to testosterone levels in
the peripheral plasma (Berndtson et al., 1974).
Mean number of mounts/first ejaculate was
1.8 compared to 1.7 for second ejaculates
obtained approximately 1 hr later (P>.05). The
high and low months for both first and second
ejaculates followed similar patterns (figure 3),
and fewer mounts were required during the
breeding season. This must be considered when
examining a stallion for breeding soundness
during the nonbreeding season.
Simple correlations between plasma testos-
terone concentrations and the other reproduc-
tive characteristics were highly significant in
many cases, but no correlation coefficient was
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623
sufficiently large (r = .50 or less) to permit
accurate prediction of reproductive character-
istics from testosterone levels in plasma sam-
ples. These results were unexpected since the
seasonal changes in many of these character-
istics appeared to parallel the seasonal changes
in the plasma testosterone levels of these
(Berndtson et al., 1974) and other stallions
(Weisner an d Kirkpatrick, 1975).
The magnitude of these correlations does
not negate the possibility that seasonal changes
in reproductive characteristics are modulated
via alterations in testosterone secretion. Rather,
the lack of meaningful correlations may have
resulted because: 1) levels for many of these
variables, including testosterone in plasma, en-
compass a rather broad range among stallions
with " n o r m a l " reproductive function, 2 ) w i d e
fluctuations in many of these variables occur at
relatively brief intervals, 3) other androgens
may be important in regulating reproductive
function, and 4) the various target organs may
require only threshold levels of testosterone for
normal function, which may have been ex-
ceeded in all stallions studied.
Episodic secretion of testosterone occurs in
the males of many species (Katongole et al.,
1971; Bartke et al., 1973). In the present study,
as much as a 4.5-fold difference in plasma levels
of testosterone was observed among samples
taken from an individual stallion within a given
month (Berndtson et al., 1974). Further, differ-
ences of as much as 23 ng o f testosterone per
milliliter in plasma were observed among sam-
ples taken from the same stallion at 20-rain
intervals for 1 hr (W. E. Berndtson, u n p u b l i s h e d
data). Thus, a single sample of blood may not
represent the modal testosterone concentration
available to the target organs.
Many seminal and behavioral characteristics
of stallions also exhibit marked fluctuations
from one sampling period to the next. Such
changes are undoubtedly influenced by a vari-
ety of extraneous factors, such as the degree of
sexual stimulation, distractions by other ani-
mals or handlers, the temperature of the artifi-
cial vagina, etc. (Hale and Almquist, 1960;
Pickett and Voss, 1975). Although an a t t e m p t
was made to provide maximal stimulus pressure
for seminal collection and to provide a uniform
degree of sexual preparation, such factors un-
doubtedly contributed to the variation in be-
havioral and seminal characteristics.
To minimize the influence of rapid, short
term fluctuations of testosterone concentration
624 PICKETT ET AL.
and seminal a n d behavioral characteristics o n
t h e c o r r e l a t i o n c o e f f i c i e n t s , t h e values f o r each
variable were also p o o l e d w i t h i n stallions o n a
per m o n t h basis b e f o r e calculating c o r r e l a t i o n
coefficients. Correlations b a s e d o n p o o l e d
m o n t h l y values were similar t o t h e simple,
overall correlations.
A l t h o u g h m o r e f r e q u e n t sampling m i g h t
have b e e n helpful, t h e m a g n i t u d e o f b o t h t h e
simple c o r r e l a t i o n c o e f f i c i e n t s a n d c o r r e l a t i o n s
calculated o n a per m o n t h basis was u n d o u b t -
edly r e d u c e d because stallions w i t h t h e h i g h e s t
c o n c e n t r a t i o n o f t e s t o s t e r o n e did n o t always
rank m o s t favorably in t e r m s o f r e p r o d u c t i v e
characteristics. F o r e x a m p l e , t h e stallion with
t h e highest overall m e a n t e s t o s t e r o n e level had
t h e longest r e a c t i o n t i m e a n d t h e lowest n u m -
ber o f s p e r m a t o z o a per ejaculate. Conse-
q u e n t l y , c o r r e l a t i o n c o e f f i c i e n t s were also cal-
culated o n a within-stallion basis. Within-stal-
lion c o r r e l a t i o n s w o u l d r e f l e c t t h e e x t e n t t o
w h i c h seasonal changes in t h e p l a s m a t e s t o s t e r -
o n e levels o f an individual stallion r e s e m b l e d
c o r r e s p o n d i n g changes in seminal a n d behav-
ioral characteristics. A l t h o u g h it was s u s p e c t e d
t h a t w i t h i n - s t a l l i o n c o r r e l a t i o n s w o u l d be high-
er t h a n t h e simple correlations, t h e values were
generally similar.
To our k n o w l e d g e , plasma t e s t o s t e r o n e lev-
els o f infertile stallions have n o t b e e n r e p o r t e d .
A l t h o u g h e x t r e m e l y high or l o w t e s t o s t e r o n e
levels m i g h t be indicative o f r e p r o d u c t i v e dis-
orders o f e n d o c r i n e origin, t h r e e m e a s u r e m e n t s
o f the plasma t e s t o s t e r o n e c o n c e n t r a t i o n per
w e e k a p p e a r o f little value in p r e d i c t i n g t h e
relative r e p r o d u c t i v e c a p a c i t y o f n o r m a l stal-
lions.
LITERATURE CITED
Bartke, A., R. E. Steele, N. Musto and B. V. Caldwell.
1973. Fluctuations in plasma testosterone levels in
adult male rats and mice. Endocrinol. 92:1223.
Berndtson, W. E., B. W. Pickett and T. M. Nett. 1974.
Reproductive physiology of the stallion. IV. Sea-
sonal changes in the testosterone concentration of
peripheral plasma. J. Reprod. Fertil. 39:115.
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 Acta Scientiae Veterinariae, 2019. 47(Suppl 1): 409.

CASE REPORT ISSN 1679-9216

Pub. 409

Testicular Teratoma in a Unilateral Right-Sided Abdominal Cryptorchid Horse

Lays Wouters Ugolini1, Fernanda Carlini Cunha dos Santos1,2, Gabriela Vicensi da Costa1, Henrique Ramos Oliveira1,
Natália Folchini1, Tanise Policarpo Machado3, Ricardo Zannella1 & Leonardo Porto Alves1

ABSTRACT

Background: Cryptorchidism is characterized by the incomplete descent of one or both testicles to the scrotum, being
a hereditary alteration and frequently an unilateral condition. Besides the sexual and aggressive behaviour, the retained
testicle is commonly located in abdominal cavity, being considered a risk factor for neoplasm development. The most
common testicular neoplasm reported in mammalian species are Sertoli cell tumors, Leydig cell tumors, seminomas and
teratomas. A presumptive diagnosis of testicular tumor can be achieved by ultrasonography, although the definitive di-
agnosis is obtained only by histopathology. In this report, we are presenting a case of testicular teratoma in an unilateral
abdominal cryptorchid horse.
Case: A 3 year-old stallion, American Quarter Horse, was attended and presented a right testicle retained inside the abdo-
men and a left testicle in the scrotum. Transrectal palpation was used to identify a round and firm structure, presumably
the right testicle, lateral to the urinary bladder and located inside abdomen. Further, a transrectal ultrasound examination
showed a complex, round mass with irregular edges containing both cystic and solid structures, hypoechoic fluid-filled
cavities separated by linear hyperechoic septa. After a clinical examination, the animal was diagnosed with cryptorchidism
and was submitted to orchiectomy and cryptorchidectomy by inguinal approach. Surgery was performed under general
anesthesia and postoperative care included cold shower, anti-inflammatory and antibiotic therapy. Testicles were surgically
removed and further sent for histopathological examination. The visual appearance of the right undescended testicle showed
multiple round, cystic, and solid structures on outer surface, while the left descended testicle was apparently normal. The
macroscopic evaluation showed that the affected testicle consisted of a firm solid mass with multiple fluid-filled cystic
areas. Microscopically, the testicular architecture was replaced by cysts, fibrous tissue, adipose tissue, glandular structures,
and foci of calcification. The histology revealed that the retained testicle had a testicular teratoma.
Discussion: Reproductive disorders are common in horses and represent a significant part of the equine practitioner
routine. Equine cryptorchidism is the most common non-lethal developmental defect of stallions. Surgery is the best
treatment, since this alteration is hereditary. Teratomas have been reported more often in cryptorchid testicles, being usu-
ally just diagnosed as an incidental finding during surgical procedure. Under field conditions, usually the testicles are not
sent for histopathological evaluation and this fact can contribute to underdiagnoses. Ultrasonography allows clinicians to
determine testis location, morphological changes in the testes, as well as to elaborate a presumptive diagnose of testicular
neoplasm. Histopathology is the best exam to achieve definitive diagnoses in the presence of testicular alterations. In our
report, diagnosis of testicular neoplasia was not made before surgery and testicular mass was an incidental finding during
pre-surgical examination. In the presence of testicular enlargement or presence of testicular mass, neoplasia should be
included in the differential diagnosis. In conclusion, although rare, teratoma should be included in differential diagnoses
of retained testicles, especially those with morphological alterations.
Keywords: andrology, neoplasm, cryptorchidism, stallion, testis.

DOI: 10.22456/1679-9216.93609
Received: 5 April 2019 Accepted: 8 July 2019 Published: 7 August 2019
1
Setor de Grandes Animais, Hospital Veterinário & Laboratório de Patologia Animal, Universidade de Passo Fundo (UPF), Passo Fundo, RS, Brazil. & 3Pro-
2

grama de Pós-graduação em Medicina Animal: Equinos, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, RS. CORRESPONDENCE:
F.C.C. Santos [carlini@portoweb.com.br - Tel.: +55 (54) 3316-7486]. Setor de Grandes Animais, Hospital Veterinário - UPF. Bairro São José. Rodovia BR
285, Km 292 s/n. CEP 99052-900 Passo Fundo, RS, Brazil.

1
L.W. Ugolini, F.C.C. Santos, G.V. Costa, et al. 2019. Testicular Teratoma in a Unilateral Right-Sided Abdominal Cryptorchid Horse.
INTRODUCTION
In horses, testicular descent generally occurs
30 days prior to and up to 2 weeks after birth [4], when
there is an incomplete descent of one or both testicles,
the animal is considered a cryptorchid [15,18]. Cryp-
torchidism is a hereditary disorder with moderated to
high heritability of 0.12 to 0.32 in horses [7] and fre-
quently unilateral [6,18]. Cryptorchidism is common in
companion animals, pigs, humans (2-12%) [1], cattle
and sheep (1%) [1], stallions (2-8%) [6,13,14].
Besides the sexual/ aggressive behavior of
cryptorchid animals, this condition is considered a
risk factor for testicular tumors development when the
undescended testes is inside the abdomen [13,14,17].
Testicular neoplasms are usually benign [14], have
been seen in descended and undescended testes [5,12],
are asymptomatic [3] and can and rarely cause abdo-
minal pain [6]. The most common tumors reported in
mammalian species are Sertoli cell tumors, Leydig cell
tumors, seminomas and teratomas [14,15,17]. Terato-
ma is quite rare in horses [2,5], however, it is the most
frequent testicular tumor reported in young stallions,
between 1 and 5 years-old [14,15,17].
Ultrasonography is a useful tool for the diagnosis
of abdominal cryptorchidism in horses [3,5]. A presump-
tive diagnosis of testicular tumor can be achieved by ultra-
sonography, however, a definitive diagnosis of testicular
teratoma can only be obtained with histopathology [3].
The aim of this study is to report a case of tes-
ticular teratoma in an unilateral right-sided abdominal
cryptorchid horse, with the description of ultrasono-
graphy evaluation, surgical approach, macroscopic and
microscopic alterations in the testis.
CASE
A 3-year-old, 350 kg, American Quarter Horse
was admitted to the Veterinary Hospital at the Universida-
de de Passo Fundo, Brazil, for elective surgical castration.
The animal presented excessively unruly, masculine
behavior, and had no history of a previous castration.
On visual inspection, the left testis was loca-
ted inside the scrotum, however no right testicle was
located. A protocol was followed in order to conduct a
clinical diagnosis of cryptorchidism: visual assessment
of the scrotum; palpation of the scrotum; palpation of
the inguinal rings; transabdominal ultrasonography
of the inguinal rings with a convex probe, transrectal
palpation; and transrectal ultrasonography.
2
Acta Scientiae Veterinariae. 47(Suppl 1): 409.
The right testicle was not found during visual
inspection and palpation. Transabdominal ultrasono-
graphy confirmed the presence of a normal left testicle
and the absence of the right testicle in the scrotum or
within inguinal region. Transrectal palpation was used
to identify a round and firm structure, presumably the
right testicle, lateral to the urinary bladder and located
inside the abdomen. Further, a transrectal ultrasound
examination showed a complex, round mass with irre-
gular edges containing both cystic and solid structures,
hypoechoic fluid-filled cavities separated by linear
hyperechoic septa.
Based on clinical examination and ultrasono-
graphy, the horse was diagnosed as cryptorchid.
Treatment was based on cryptorchidectomy
and orquiectomy. For surgical removal of the testicles,
the animal underwent general intravenous anesthesia.
Before the surgery, a combination of procaine penicillin,
benzathine penicillin, dihydrostreptomycin sulfate and
piroxicam1 (20.000 UI/kg IM) was administered. Xyla-
zine2 (1.1 mg/kg IV) was the premedication used, and
induction was performed with a combination of diaze-
pam3 (0.04 mg/kg IV) and ketamine2 (2.2 mg/kg IV). A
constant rate infusion (0.05 mL/kg/min) of a combination
of eterglicerilguaiacol4 (50 mg/mL), lidocain without
vasoconstrictor5 (250 mg), ketamine2 (2 mg/mL) and
xylazine2 (1 mg/mL) was maintained during surgery
Before the testes were surgically removed, intratesticular
anesthesia with lidocaine5 was used. During the surgery,
the patient received a single dosis of anti-tetanus serum6
(5.000UI IM) and flunixin meglumine1 (1.1 mg/kg IV).
The horse was placed in dorsal recumbence, and
the area was aseptically prepared with 2% chlorhexidine
detergent and 0.5% chlorhexidine alcohol solutions. The
surgical technique for cryptorchidectomy by inguinal
approach was conducted as described by Hendrickson
[10], except for the use of a grasping forceps. The guber-
naculum was not found, and the abdominal cavity was
manually inspected until the right undescended testicle
was found. The contralateral left testis was removed by
orchiectomy as described by Hendrickson [10]. Testicles
were referred to histopathological evaluation.
Postoperative care included flunixin meglu-
mine1 (1.1 mg/kg IV q24 h) for 5 days, association of
procaine penicillin, benzathine penicillin, dihydros-
treptomycin sulfate and piroxicam1 (20.000 UI/kg IM
q48 h) for 3 times, and cold showers for 20 min (2x a
day) for 10 consecutive days.
L.W. Ugolini, F.C.C. Santos, G.V. Costa, et al. 2019. Testicular Teratoma in a Unilateral Right-Sided Abdominal Cryptorchid Horse.
Acta Scientiae Veterinariae. 47(Suppl 1): 409.

The macroscopy of the testicles showed that

Figure 1. a- Descended left testis. b- Cryptorchid, undescended right testis.
the left testicle was apparently normal (Figure 1a)
while the right testis was round, had cystic and solid
structures detected on palpation (Figure 1b).
Left testis measured 13.0 x 6.0 x 5.3 cm. There
was edema of the tail of the epididymis and no altera-
tions in the testicular parenchyma on cut surface (Figure
2a). Right testis measured 8.0 x 6.5 x 6.3 cm, was firm,
had multiple cysts with brownish fluid, with calcified
areas alternating with firm white areas (Figure 2b).
Both testes were submitted for histopathologi-
cal examination. No microscopic lesions were found in
the left testis, which had preserved spermatogenesis.
Parenchyma of the right testis was necrotic and had de-

Figure 2. a- Descended left with no alterations in the testicular parenchyma on cut surface. b- Cryptorchid right testis. Testicular architecture is obliter-
ated by the teratoma (red circle indicates cystic formations).

Figure 3. Microscopic findings in the retained testis. a- Degenerated glandular tissue [Obj.×400; Bar= 40 µm]. b- Presence of bone trabeculae (bone
tissue) (asterisk). [Obj.×200; Bar= 100 µm].

3
L.W. Ugolini, F.C.C. Santos, G.V. Costa, et al. 2019. Testicular Teratoma in a Unilateral Right-Sided Abdominal Cryptorchid Horse.
generated glandular structures (Figure 3a), fibrous tissue,
cystic areas alternated with bone trabeculae (Figure 3b),
adipose tissue, areas of calcification, moderate multifocal
hemorrhage, and mixed inflammatory infiltrate. A diag-
nosis of testicular teratoma was made by histopathology.
DISCUSSION
Reproductive disorders are common in horses
and represent a significant part of the equine practi-
tioner work load [8]. Equine cryptorchidism is the
most common non-lethal developmental alteration
in stallions and surgical removal of testes is the best
treatment, since the hereditary aspect and the risk of
neoplasm development.
Abdominal cryptorchid humans have higher
risk of developing testicular neoplasms than inguinal
cryptorchid patients. The relative position of the cryp-
torchid testis and the severity of environmental insults
on the gonads, e.g. heat, likely play a role in the occur-
rence and prevalence of these neoplasms [6]. Precursor
cancer cells are generally pluripotent germ cells, which
are derived from primordial germ cells, that continue
to proliferate or undergo improper differentiation [16].
The presence of an abnormal testis dramatically alters
the function of somatic cells, providing the niche for
spermatogonial steam cells self-renewal and diffe-
rentiation. In humans, a possible explanation is that
the ectopic testis itself is directly responsible for the
infertility and germ cell tumorigenesis [13]. Althou-
gh rare, teratomas have been reported more often in
cryptorchid testicles inside abdominal cavity [12] and
occurs sporadically in young stallions [3,6,8,12,14,15],
similar to present case report.
In the present case, careful clinical examination
of the stallion allowed to find the retained, undescen-
ded testes, located intra-abdominal. During physical
examination, the testes are carefully palpated, and
the size, symmetry and consistency of the gonad and
surrounding structures are the parameters assessed.
Inguinal and transrectal palpation are reliable tools to
locate the testes [11]. If there is any abnormality during
the physical examination, then the use of ultrasound
should be considered. In our case, morphological
changes were detected in one of the testis during
transrectal ultrasound, but we were unable to accura-
tely identify this structure as testis. Ultrasonography
allows clinicians to determine testis location as well as
morphological changes in the testicular parenchyma,
4
Acta Scientiae Veterinariae. 47(Suppl 1): 409.
such as enlargement of the gonad possibly caused by
neoplasia [3,9]. As ultrasound imaging depicts the
morphology of the examined tissues, veterinarians may
be able to narrow down the diagnosis significantly [3].
If more information is needed, then a testicular biopsy
should be performed and the sample submitted for
histopathological evaluation to provide a definitive
diagnosis. Histopathology analysis is more accurate
than macroscopic inspection and clinical examination
of the testis.
In the present study, a presumptive diagnosis
of testicular tumor was made based on the ultrasound
findings and macroscopic appearance of the surgi-
cally removed testis. Seminoma, Sertoli cell tumor,
Leydig cell tumor, teratocarcinoma, and embryonal
carcinoma are the testicular neoplasms that should be
included in differential diagnoses of teratoma [5]. The
definitive, final diagnosis, in our case was based on
the microscopical findings in the surgically removed
undescended testis.
Teratomas are formed by various different
types of mature tissues originating from different ger-
minal layers and which embryonic origin differs from
the affected organ [5,12,14,17]. Tumor size varies gre-
atly and many are cystic or polycystic [15], and usually
grey to yellow [14,15]. Usually the retained testis is
smaller than the scrotal testis. In our case, fibrous tis-
sue, cysts, bone, adipose tissue, and mineralized areas
were found by light microscopy. A variety of tissues
can be identified in teratomas including skin, muscle,
tooth, hair follicles, bone, and cartilage [2,12,14,15].
The actual prevalence of testicular teratoma in global
equine population is difficult to estimate without large
retrospective and prospective studies, because most
stallions are castrated at an early age. Since testicular
teratomas rarely cause clinical signs, the tumor is of-
ten detected during preoperative assessment, during
surgery or at necropsy, as an incidental finding. Under
field conditions, few veterinarians submit the retained
undescended surgically removed testes for histopatho-
logy, unless when significant morphological alterations
are noted. Possibly these factors contribute to the
underdiagnoses and missdiagnoses of this neoplasm.
In our report, diagnosis of testicular neoplasia
was not made before surgery and testicular mass was an
incidental finding during the pre-surgical examination.
Before testicular enlargement or presence of testicular
mass, neoplasia should be included in the differential
L.W. Ugolini, F.C.C. Santos, G.V. Costa, et al. 2019. Testicular Teratoma in a Unilateral Right-Sided Abdominal Cryptorchid Horse.
Acta Scientiae Veterinariae. 47(Suppl 1): 409.

diagnosis. Testicular teratomas are rarely reported in MANUFACTURERS

the veterinary literature and possibly underdiagnosed 1

MSD Saúde Animal. São Paulo, SP, Brazil.
2
Vetnil Ind. e Com. de Produtos Veterinários Ltda. Louveira, SP,
in cryptorquid stallions. Brazil.
3
União Química Farmacêutica Nacional. São Paulo, SP, Brazil.
Declaration of interest. The authors report no conflicts of 4
JA Saúde Animal. Patrocínio Paulista, SP, Brazil.
interest. The authors alone are responsible for the content and 5
Novafarma Indústria Farmacêutica Ltda. Anapolis, GO, Brazil.
writing of the paper. 6
Venco Saúde Animal. Londrina, PR, Brazil.

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5
 CASE STUDY
Equine Testicular Lesions Related to Invasion
by Nematodes
G. Marino,a A. Zanghi,a M. Quartuccio,a S. Cristarella,a M. Giuseppe,a
and G. Catoneb
ABSTRACT
Nematodes of the genuses Strongylus and Setaria are
able to erratically migrate to the equine testis and pro-
voke characteristic inflammatory changes. Moreover,
the presence of living parasites in scrotal testes is a scarce
finding in which only the tracts and related lesions may
be observed. In a 20-year experience of equine testes
observed in abattoirs, 13 cases of atypical inflammatory
lesions in scrotal testes were collected. On opening of
the vaginal cavity, hydrocele was consistantly observed,
and the Morgagni’s appendix was enlarged, prominent,
and reddish. Large, elongated, or irregular prominent
red to yellow areas were visible on the testicular or epi-
didymal serosa. Edematous enlargement of the epididy-
mal head or tail was occasionally observed. In two cases,
the described lesions were detected together with nem-
atodes of the species Setaria equina, free and viable or
encapsulated in the visceral and parietal layers of the
tunica vaginalis, confirming the hypothesized cause.
Histologically, the serosal lesions were interpreted as
parasitic tracts, characterized by hemorrhage and leuko-
cyte infiltrates in recent lesions or by granulation tissue
with numerous macrophages in elderly ones. In the lat-
ter, areas of squamous metaplasia could be observed in
the appendix testis and in the epididymal duct. Associ-
ated microscopic lesions included diffuse periorchitis,
lymphocytic interstitial orchitis with mild to severe
degeneration of seminiferous tubules, and vasculitis,
epididymitis, and sperm granuloma.
Keywords: Stallion; Scrotal testes; Nematode; Patho-
logic lesions
INTRODUCTION
Among strongyloidae, larvae of Strongylus edentatus and
adults of Setaria equina have been occasionally reported
From the Department of Veterinary Public Health, University of Messina, Messina,
Italy; and the a Department of Veterinary Science, University of Camerino,
Camerino, Italy.b
Reprint requests: Mazzullo Giuseppe, Department of Veterinary Public Health,
University of Messina, Messina, Italy.
0737-0806/$ - see front matter
Crown Copyright Ó Published by 2009 Elsevier Inc. All rights reserved.
doi:10.1016/j.jevs.2009.08.004
728
in the genital organs of the stallion. They may be detected
in the testicular parenchyma and epididymis of retained tes-
tes and, rarely, in the vaginal cavity of scrotal gonads, par-
ticularly in young horses.1-4 It has been hypothesized that
parasites reach the testis through the spermatic artery or di-
rectly from the peritoneum, moving under the tunica vag-
inalis of the testicular cord.1,3,5 If the parasite dies in these
aberrant districts, a granulomatous reaction develops, as
demonstrated in the bull by an experimental infection of
testes with dead larvae of Setaria labiatopapillosa.6 Most
infestations are not clinically manifested, with the excep-
tion of massive ones that cause an increase in testicular
size.7 In addition, Strongylus spp larvae have been indicated
as the causative agent in cases of acute orchitis, epididymi-
tis, and sperm granuloma.3,7,8 In one study, Strongylus spp.
larvae were detected in the testes of 11 stallions between 2
and 7 years old.5 One was bilaterally cryptorchid with lar-
vae in both testes, whereas the others had only one affected
gonad, which was abdominal in seven cases, inguinal in
two, and scrotal in one case. The larvae and their tracts
were observed in the testicular cord and in the testis,
from the tunica albuginea to the center of the parenchyma.
The tracts consist of channels approximately 2 mm wide
containing leukocytes, with many eosinophils, and hemor-
rhage that may extend to adjacent parenchyma.3 There are
often no other signs of the parasite invasion in scrotal tes-
tes.2 Sometimes, parasites invade the appendix testis, caus-
ing hemorrhagic lesions, severe inflammation, and foci of
squamous metaplasia (SM) of the ductal epithelium.2
Even some cases of focal lymphocytic orchitis associated
with arteritis and testicular degeneration of seminiferous
tubules have been related to migration of Strongylus spp.
larvae.2,3 This study aimed to describe some testicular
lesions related to invasion by nematodes.
MATERIALS AND METHODS
In a 20-year experience of equine testes collected at slaugh-
ter or castration, 13 scrotal testes of 1- to 7-year-old
(mean, 5  2 years) stallions of different breeds were
selected. The period of collection ranged from February
to July. Criteria for selection included presence of atypical
inflammatory changes on the serosa and near the appendix
testis associated or not with free or encapsulated worms.
Testes were grossly examined, and specimens from the
Journal of Equine Veterinary Science  Vol 29, No 10 (2009)
G Marino et al  Vol 29, No 10 (2009) 729

Table 1. Testicular gross lesions observed in the case series

N Testis Periorchitis Appendix Involvement Serosal Tracts Testicular Tracts Epididymal Changes Parasites
1 Right Y Y Y
Left Y Y Y
2 Right Y Y Y
3 Right Y Y Y
Left Y Y Y
4 Right Y Y
Left Y Y Y
5 Right Y Y Y Y Y
Left Y Y
6 Right Y Y Y
7 Right Y Y Y
8 Right Y Y Y
9 Right Y Y
Left Y Y
10 Left Y Y
11 Left Y Y
12 Right Y Y Y Y
Left Y Y
13 Right Y Y Y Y
Left Y Y Y Y

Fig. 1. Prominent reddish formation involving the Fig. 2. Hemorrhagic tracts (arrows) in the testicular
appendix testis in relation to two specular testicular and parenchyma at section.
epididymal tracts (arrow).

with a moderate amount of yellow to pinkish fluid was ob-

testis, epididymis, and testicular cord were fixed in 10%
buffered formalin, paraffin-wax embedded, and stained
with hematoxylin-eosin. When free worms were detected,
they were fixed in 70% ethanol, clarified in lactophenol,
and identified under a light microscope.
RESULTS
The observed gross lesions are summarized in Table 1. On
opening the vaginal cavity of the affected testes, hydrocele
served. The parietal and visceral tunicae vaginalis were
thickened, and several fibrinoid or fibrous adhesions were
present. Large (up to 3–4 cm), elongated or irregular,
bloody or orange to yellowish, and slightly prominent areas
could be observed on the surface of the testis and epididy-
mis, particularly at the cranial pole. The appendix testis ap-
peared enlarged, hypertrophic, and congested, occasionally
included in the serosal lesions described (Fig. 1). The epi-
didymal head and tail could be enlarged with gelatinous
730 G Marino et al  Vol 29, No 10 (2009)

Fig. 3. Irregular hemorrhagic areas on the visceral

tunica vaginalis associated with a free nematode (Setaria
equina) (arrow).

Fig. 4. Two parasites (Setaria equina) encapsulated in

edema. The sectioned testes were mostly unaffected or
contained mild fibrotic areas; in one case, 2-mm hemor-
rhagic channels were observed in different areas of the tes-
ticular parenchyma (Fig. 2). In two cases, comparable
lesions were detected together with nematodes. In the first
case, three free adult worms were collected from the vagi-
nal cavity (Fig. 3) associated with periorchitis with elon-
gated hemorrhagic areas and edematous enlargement of
epididymis. In the second case, two adult worms were lo-
cated under the visceral layer of the right testis surrounded
by a reddish area (Fig. 4), whereas a third worm was encap-
sulated in the parietal tunica vaginalis of the contralateral
testis. The inflammation of the appendix testis was clearly
detectable in both testes. On the basis of morphologic
characteristics, the parasites were identified as S. equina.4
The histologic findings are summarized in Table 2. The tu-
nicae vaginalis showed proliferation of connective tissue
with abundant smooth muscle fibers and included several
microgranulomatous foci consistent with chronic inflam-
mation. The serosal lesions and the testicular channels ap-
peared as hemorrhagic tracts in a developing granulation
tissue. Acute lesions were characterized by abundant hem-
orrhage invading surrounding structures. Frequent vascu-
litis and perivasculitis were observed together with
infiltrates of lymphocytes, hemosiderin-filled macro-
phages, mast cells, and eosinophils. The chronic lesions,
orange to yellowish, were made up of perfused fibrous con-
nective tissue with modest infiltrates of macrophages cov-
ered by a metaplastic multilayer epithelium (Figs. 5 and
6). When parasite bodies were found encapsulated in the
tunics, a large granulomatous reaction developed that in-
cluded epithelioid cells and numerous eosinophils (Figs.
7 and 8) The appendix testis during acute inflammation
the visceral tunica vaginalis associated with chronic
periorchitis.
had a hemorrhagic stroma with inflammatory infiltrates.
Chronic lesions included hyperplasia or SM of the epithe-
lium of its tubular structures (Fig. 9). In most cases, the
testicular parenchyma displayed normal spermatogenesis;
however, mild to severe degeneration, peritubular edema,
and focal interstitial lymphocytic infiltrates (focal lympho-
cytic orchitis) could be observed (Fig. 10). The epididymis
showed mild to severe lesions, such as interstitial edema,
inflammatory infiltrates, sperm granulomas, hyperplasia,
and cysts of the ductal epithelium. In case 12, SM of the
epididymal epithelium was observed, with lesions from
focal to diffuse releasing keratin debris into the lumen
(Fig. 11).
DISCUSSION
The presence of nematode species S. edentatus and S.
equina has been described in retained testes but rarely in
scrotal ones.2,4,5 The parasites have limited access to the
vaginal cavity because of the distance, the narrow inguinal
rings, and the unsuitable temperature. For these reasons, in
scrotal testes only traumatic injuries or granulomatous
lesions may be detected2,3 as signs of the transitory invasion
of these nematodes or as a consequence of their death in
this aberrant district. Despite the absence of worms, the
traumatic or phlogistic changes occurring in 11 cases are
too specific and unlikely to be attributable to another
cause, as suggested in other reports.2,3 In our cases, the de-
tection of comparable lesions in conjunction with parasites
G Marino et al  Vol 29, No 10 (2009) 731

Table 2. Histopathologic findings in the case series

Appendix Testis Tracts Testis
N Testis Acute Chronic Acute Chronic Degeneration Orchitis Epididymitis
1 Right Y Y Y Y
Left Y Y Y
2 Right Y Y
3 Right Y Y
Left Y Y
4 Right Y
Left Y Y
5 Right Y Y Y
Left Y
6 Right Y Y Y
7 Right Y Y
8 Right Y Y
9 Right Y
Left Y
10 Left Y Y
11 Left Y
12 Right Y Y Y Y Y
Left Y Y Y
13 Right Y Y Y Y
Left Y Y Y Y

Fig. 6. Transitional area (arrow) between the

unaffected testicular albuginea and the organized tract
Fig. 5. An elderly tract with SM (left) close to the covered by a metaplastic epithelium. HE, original
appendix testis (right). HE, original magnification magnification 100.
200.

confirmed the close relationship of the two events. The enclosing the Morgagni’s appendix, reveals the tendency
shape of the tracts, often elongated, could be related to of these worms to penetrate its tubular structures.2 The
the morphology of the parasites. The location of the tracts, hemorrhagic areas within granulation tissue indicate a me-
generally at the cranial pole of the testis and sometimes chanical trauma at the time of penetration and transit of the
732
Fig. 7. Granulomatous reaction developed around
a parasitic body in the wall of tunica albuginea. HE,
original magnification 100.
Fig. 8. A parasitic granuloma in the parietal tunica
vaginalis. HE, original magnification 100.
parasites. The type of inflammatory infiltrate, which in-
cludes many lymphocytes, eosinophils, and mast cells,
characterizes these lesions immunologically, demonstrat-
ing the biologic nature of the trauma. The SM, frequently
found in the tubular structures of Morgagni’s appendix
and superficially in the organized tracts, has no other path-
ogenetic interpretation in report. This is a slowly evolving
lesion, but it is likely that a massive parasitization may cause
severe phlogosis, and abnormal temperature and clinical
signs are detectable.7,8 Invasion of the tunica vaginalis
causes a periorchitis; the inflammatory process may extend
to the testis, leading to focal lymphocytic orchitis, degener-
ation of seminiferous tubules, and vasculitis, and to the
G Marino et al  Vol 29, No 10 (2009)
Fig. 9. SM of deep tubular structures of the appendix
testis. HE, original magnification 200.
Fig. 10. Interstitial lymphocytic orchitis and testicular
degeneration. HE, original magnification 100.
epididymis, leading to epididymitis and sperm granu-
loma.2,3 The SM of the epididymal duct observed in one
case, with no precedent in the literature, could be another
pathologic feature of this parasitic invasion. In our series,
we identified S. equina as the nematode mainly responsible
for periorchitis and serosal tracts in the equine testis. S.
edentatus may be involved in the tracts specifically localized
adjacent to the appendix testis and into the testicular pa-
renchyma, as indicated in report,2,3 but no worms belong-
ing to this species were found in this study. On the basis of
these findings, pathologists have to consider the occasional
migration of nematodes as causative agents of atypical asso-
ciations of periorchitis, orchitis, or epididymitis with recent
G Marino et al  Vol 29, No 10 (2009)
Fig. 11. SM of epididymal duct epithelium with keratin
debris in the lumen. HE, original magnification 200.
or already organized traumatic lesions. These could be mis-
diagnosed as appendix torsions because of the frequent in-
volvement of this structure, or as squamous cell carcinomas
because of the commonly detected SM.2
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