ARBOVIRUS (Arthropod-borne viruses) -A summarized virus name that is transmitted by arthropods (is an invertebrate animal having an external skeleton,

segmented body, and jointed appendages. Members of the Phylum Arthropoda and include the insects, arachnids, crustaceans, and others). -are group of infectious agents that are transmitted by blood-sucking arthropods from one vertebrate host to another. -They multiply in the tissues of the arthropods without evidence of disease or damage. -The vector acquires a lifelong infection through ingestion of blood from viremic vertebrae. -Some of these are maintained in nature by transovarian and possibly sexual transmission in arthropods. -Found in temperate and tropical zone but most prevalent in tropical rain forest where animals and arthropods are abundant. -More than 450 in number; about 100 are known pathogenic to humans. -Classified according to chemical and physical properties and their antigenic relationships. - Disease produced by Arbovirus may be divided into 3 clinical syndromes: encephalitis febrile diseases (sometimes associated with rash) hemorrhagic fevers

*Classification (3 major [TOGAVIRUS, FLAVIVIRUS AND BUNYAVIRUS]) TOGAVIRUS

alphavirus subgroup consists of 30 viruses that are 70 nm in diameter and a single-stranded, positive-sense RNA genome. The envelop contains two glycoproteins and lipids. -Establish persistent infections in mosquitoes and are transmitted between vertebrates by mosquitoes. -all are antigenically related. -inactivated by acid ph, heat, lipid solvents, detergents, bleach, phenol, 70% alcohol, and formaldehyde. -Most posses hemagglutinating ability. Rubella virus is classified in a separate genus in the togaviridae family and is not an arbovirus. -replicates in the cytoplasm; mature by budding nucleocapsids through the plasma membrane. Morphology Enveloped, spherical particles, 65-70nm diameter. Capsid: 240 monomers, icosahedral, Envelope: 80 trimer spikes Antigenic properties - All alphaviruses are antigenically related. Because of common antigenic
determinants, the viruses show cross-reactions in immunodiagnostic techniques. - HI, ELISA, and IF tests define eight antigenic complexes or serogroups of alphaviruses, four of which are typified by western equine encephalitis, eastern equine encephalitis, Venezuelan equine encephalitis, and Semliki Forest virus. - Identification of a specific virus can be accomplished using Nt tests.

Pathogenesis -Virus is transmitted from the salivary glands of the mosquito to the bloodstream of the vertebrate host. Virus travels to the skin and reticuloendothelial system (spleen and lymph nodes), where the primary infection occurs, then viraemia follows - systemic infection. Can involve CNS (esp. encephalitis), skin/bone marrow/blood vessels (haemorrhagic fevers). Infectious Cycle

1. Attachment- attachment of viral capsids/spikes and protein to specific receptor to host cell. 2. Penetration- force/endocytosis or membrane fussion. 3. Uncoating- capsid is being removed and dissolved through host cell enzymes leaving only the genetic material. 4. Biosynthesis- synthesis of viral messenger RNA/viral protein synthesis or assembly of viral protein and viral genom. 5. Release

Laboratory Diagnosis 1. Culture can be grown in both vertebrate and mosquito cell lines but difficult to isolate 2. Cytopathologic studies 3. Immunofluorescence 4. Reverse transcriptase-PCR 5. Serology hemagglutination inhibition, ELISA, latex agglutination Presence of specific IgM or 4-fold rise in titer between acute and convalescent sera indicate a recent infection

Examples of virus under Togaviridae family: Eastern Equine Encephalitis -EEE is a member of the antigenically similar family of viruses known as Togaviridae, which also includes western equine encephalitis and Venezuelan equine encephalitis. These alphaviruses are spherical and have a diameter of 60-65 nm. The outer layer consists of a glycoprotein shell with protruding glycoprotein spikes found beneath the lipid bilayer. The nucleocapsid core contains the single-stranded RNA genome.

Pathogenesis EEE is characterized by diffuse CNS involvement. A large number of immunologically active cells enter the brain parenchyma and perivascular areas and mediate much of the damage. Infiltrating neutrophils and macrophages cause neuronal destruction, neuronophagia, focal necrosis, and spotty demyelination. Vascular inflammation with endothelial proliferation, small vessel thrombosis, and perivascular cuffing may also develop. Antigenic studies reveal that EEE primarily affects the perikaryon and dendrites of neurons, with minimal findings in glial cells. Occasionally, secondary glial proliferation and the formation of glial nodules occur. Cell death by apoptosis occurs primarily among the glial and inflammatory cells. Gross inspection on autopsy reveals edema, leptomeningeal vascular congestion, hemorrhage, and encephalomalacia. Patients who die late in the disease may exhibit diffuse cerebral atrophy, particularly of the cortex. The mosquito injects the agent of EEE into the subcutaneous and cutaneous tissues of the host. EEE is not transmitted via the aerosol route. It may cross the placenta and infect the fetus. Because of low viral titers in the donor's blood, EEE is unlikely to be transmitted via transfusion. The prodrome of fevers, chills, weakness, headache, and myalgias represents replication of the virus in nonneural tissues (tissue adjacent to the mosquito's bite or the lymphatic system). The virus then binds to specific tissue receptors, undergoes endocytosis, and initiates an RNA-dependent RNA and protein synthetic process. If the original inoculum is large enough, secondary viremia occurs, with eventual viral migration into the CNS via cerebral capillary endothelial cells. Poorly described features of the virus increase microvascular permeability of the brain. Cell-to-cell spread then occurs via dendrites and axons. These initial symptoms often progress rapidly to confusion, somnolence, or even coma. Effects on human - Fatigue -Fever -Headache -Nausea -Restlessness or irritability -Difficulty walking or unstableness -Confusion, impaired judgment, or an altered mental state -Seizures Infectious Cycle

6. Attachment- attachment of viral capsids/spikes and protein to specific receptor to host cell. 7. Penetration- force/endocytosis or membrane fussion. 8. Uncoating- capsid is being removed and dissolved through host cell enzymes leaving only the genetic material. 9. Biosynthesis- synthesis of viral messenger RNA/viral protein synthesis or assembly of viral protein and viral genom. 10. Release

Incubation Period Incubation period is between 3-10 days. Laboratory Diagnosis Laboratory diagnosis of arboviral infections is generally accomplished by testing of serum or cerebrospinal fluid (CSF) to detect virus-specific IgM and neutralizing antibodies. During an acute infection, certain viruses can be isolated through culture or detected by nucleic acid amplification. In fatal cases, nucleic acid amplification, histopathology with immunohistochemistry and virus culture of autopsy tissues can also be useful. Only a few state laboratories or other specialized laboratories, including those at CDC, are capable of doing this specialized testing. Medication Drugs currently used are those capable of ameliorating neurologic complications. No current studies provide support for or against prophylactic use. Potentially used medications include phenytoin, phenobarbital, or a benzodiazepine drip. Use antipyretics as needed. Additionally, appropriate analgesics and amnestics can be used once the patient is intubated.

Western equine encephalitis

WEE is of the genus Alphavirus and in the family Togaviridae. WEE is a summertime infection found in the western United States, and it is more common in rural areas. WEE is a member of the antigenically similar group of viruses known as Togaviridae, which encompasses eastern equine encephalitis (EEE) and Venezuelan equine encephalitis (VEE). These alphaviruses are spherical and have a diameter of 60-65 nm. The outer layer consists of a glycoprotein shell with protruding glycoprotein spikes, beneath which lies the lipid bilayer. The nucleocapsid core contains the single-stranded RNA genome.

Pathogenesis The WEE virus is a neurotropic alphavirus, which causes encephalitis and viral symptoms without an associated rash. The disease is usually subclinical and may mimic many viral and inflammatory syndromes. Diffuse CNS involvement characterizes WEE in its more severe stages. Much of the damage is mediated by the large number of immunologically active cells that enter the brain parenchyma and perivascular areas. Focal necrosis is often found in the striatum, globus pallidus, cerebral cortex, thalamus, pons, and meninges. Neutrophils and macrophages may infiltrate the brain parenchyma and may cause neuronal destruction, neuronophagia, focal necrosis, and spotty demyelination. Vascular inflammation with endothelial proliferation, small vessel thrombosis, and perivascular cuffing may also occur. Cell death by apoptosis occurs primarily in the glial and inflammatory cells. Gross inspection during autopsy reveals edema, leptomeningeal vascular congestion, hemorrhage, and encephalomalacia. In infants or children who die of the disease, diffuse atrophy, particularly of the cortex, may be present. Pathogen invasion The virus is transmitted from the mosquito into subcutaneous and cutaneous tissue of the host. It cannot be transmitted via the aerosol route. The virus can also be transferred transplacentally. In the fetus, infection often results in massive cerebral necrosis and death. Infection via contaminated blood transfusions is unlikely because the level of viremia in the donor is extremely low. The infected individual usually develops a general viral prodrome with fevers, chills, weakness, headache, or myalgias. Viral replication in nonneural tissues, most often adjacent or lymphoid tissue, marks this period.

The virus binds to specific tissue receptors, undergoes endocytosis, and begins an RNA-dependent synthesis of RNA and protein. If the inoculum is high enough, a subsequent viremia develops, with eventual translocation to the CNS via cerebral capillary endothelial cells. The exact mechanism of this is not known but is believed to be secondary to vascular infiltration because factors that increase vascular permeability often facilitate neuroinvasion. Cell-to-cell spread in the CNS occurs via neighboring dendrites and axons. The initial symptoms may progress rapidly to CNS symptoms of mental confusion, somnolence, coma, and death in 1-2 days, or they may resolve without sequelae. During epidemics, a significant percentage of the population seroconverts, but the case-infection ratio is low in human adults (1:1000) and high in infants (1:1). Most infected individuals rarely experience severe CNS manifestations, and most infections are subclinical. An inverse ratio has been found between age and clinical CNS manifestations, including seizures and other sequelae. Effects on human Nausea Vomiting Diarrhoea Fever Tachycardia Tremors Cyanosis Seizures Increased reflexes Lymphadenopathy Malaise Rash Myalgia

Stiff neck Headache Photophobia Lethargy Coma Aphasia Anosmia Asymptomatic Infections Febrile illness Drowsiness Irritability Confusion Weakness

Infectious Cycle 1. Attachment- attachment of viral capsids/spikes and protein to specific receptor to host cell. 2. Penetration- force/endocytosis or membrane fussion. 3. Uncoating- capsid is being removed and dissolved through host cell enzymes leaving only the genetic material. 4. Biosynthesis- synthesis of viral messenger RNA/viral protein synthesis or assembly of viral protein and viral genom. 5. Release Incubation Period Incubation is between 1-5 days. Laboratory Diagnosis

Laboratory diagnosis of arboviral infections is generally accomplished by testing of serum or cerebrospinal fluid (CSF) to detect virus-specific IgM and neutralizing antibodies. During an acute infection, certain viruses can be isolated through culture or detected by nucleic acid amplification. In fatal cases, nucleic acid amplification, histopathology with immunohistochemistry and virus culture of autopsy tissues can also be useful. Only a few state laboratories or other specialized laboratories, including those at CDC, are capable of doing this specialized testing. Medication The drugs currently used consist of agents capable of ameliorating neurologic complications. Antipyretics are used as needed. Additionally, suitable analgesics and amnestics are appropriate once the patient is intubated. Initiate anticonvulsants either when a seizure has occurred or is probable, particularly in the pediatric population, in whom prevalence is high. Corticosteroids are administered early and serve multiple functions. They decrease inflammation, decrease cerebral edema, and correct any adrenocortical insufficiency. Venezuelan equine encephalitis Venezuelan equine encephalitis (VEE) is an acute viral disease characterized by fever, chills, headache, nausea, vomiting, lumbosacral pain, and myalgia, which may progress to encephalitis. It is caused by the Venezuelan equine encephalitis virus and is a significant disease in the Americas. Pathogenesis Alphaviruses are limited in their geographic spread primarily by the presence of an appropriate competent arthropod vector. At least 10 mosquito species, including Aedes, Culex, Psorophora, Mansonia, and Deinocerites species, have been identified as probable epidemic vectors for the Venezuelan equine encephalitis virus, with different mosquito vectors possessing varying levels of efficiency. The mosquito vector becomes infected after biting a viremic equine host. Humans can develop a viremia significant enough to infect mosquitos, but humans never have been directly implicated in epidemic transmission. Although Venezuelan equine encephalitis virus can be demonstrated in human throat swabs, human-to-human transmission never has been demonstrated.

For approximately one week, the virus replicates in the midgut epithelium of the mosquito. The virus then is disseminated to other organs, including the hemolymph and salivary glands. Spread to humans occurs when the infected mosquito deposits the virus in the skin of a nave host while feeding. Viremia and a febrile response mark the initial phase of infection, during which the virus replicates in extraneural tissues. Sites of human replication remain unclear, but, in equines and laboratory rodents, the sites include skeletal muscle, lymphoid, and hematopoietic tissues. This may lead to relative lymphopenia, neutropenia, and thrombocytopenia. Circulating virus gains access to the CNS via the blood stream or perhaps via the olfactory apparatus. Neuronal infection with Venezuelan equine encephalitis is associated with the onset of acute encephalitis and cell death by apoptosis. Effects on human High fever Headache Flu-like symptoms Rigors Sore muscles Sore throat Nausea Vomiting Diarrhea Confusion Swollen lymph glands Tremors Double vision Photophobia

Infectious Cycle

1. Attachment- attachment of viral capsids/spikes and protein to specific receptor to host cell. 2. Penetration- force/endocytosis or membrane fussion. 3. Uncoating- capsid is being removed and dissolved through host cell enzymes leaving only the genetic material. 4. Biosynthesis- synthesis of viral messenger RNA/viral protein synthesis or assembly of viral protein and viral genom. 5. Release Incubation Period Incubation period is 24-72 hours. Laboratory Diagnosis Those who handle infectious VEE viruses or their antigens prepared from infected tissues or cell cultures should be vaccinated and shown to have demonstrable immunity in the form of VEE virus-specific neutralising antibody. All procedures producing aerosols from VEE virus materials should be conducted in biosafety cabinets at containment level. A confirmatory diagnosis of VEE is based on the isolation and identification of the virus or on the demonstration of seroconversion. Medication No specific treatment for Venezuelan equine encephalitis (VEE) infections exists. Anticonvulsants These agents are used to prevent seizure recurrence and to terminate clinical and electrical seizure activity.

FLAVIVIRUS

-The family consists of 65 viruses that are 40-50 nm in diameter and have an enveloped, positive-sense RNA. The envelope contains a single glycoprotein and lipid.

-replicates in the cytoplasm; mature through intarcytoplasmic membranes (particularly endoplasmic reticulum). Morphology Spherical, 40-60nm; Capsid: Symmetry indistinct, 2 proteins: nucleocapsid ('C') and matrix ('M'); Envelope: 1 glycoprotein ('E').

Pathogenesis Produce a wide range of diseases - fever; arthralgia; rash; haemorrhagic fever; encephalitis). The outcome of infection is influenced by both virus and host-specific factors (age, sex, genetic susceptibility, pre-exposure to same or related agent). Laboratory Diagnosis 1. Culture can be grown in both vertebrate and mosquito cell lines but difficult to isolate

2. Cytopathologic studies 3. Immunofluorescence 4. Reverse transcriptase-PCR 5. Serology hemagglutination inhibition, ELISA, latex agglutination Presence of specific IgM or 4-fold rise in titer between acute and convalescent sera indicate a recent infection

Examples of virus under Flavivirus family: 1.St. Louis encephalitis virus (SLEV) belongs to the family Flaviviridae (group B arborviruses). The principal reservoirs of SLEV include wild birds and domestic fowl, and the virus is transmitted to humans by mosquitos (Culex tarsalis, Culex quinquefasciatus, Culex pipiens). The clinical manifestations of SLEV infection range from mild flulike syndromes to fatal encephalitis

2. The Dengue virus is a member of the virus family Flaviviridae and is transmitted to people through the bite of the mosquitos Aedes aegypti and Aedes albopictus. Dengue virus is now believed to be the most common arthropod-borne disease in the world. Dengue is mainly found in the tropics because the mosquitoes require a warm climate. A major fear of epidemiologists is that the mosquitoes will develop resistance to cooler climates and then be able to infect people in the United States and other temperate climates. The virus is transmitted when a mosquito of the Aedes genus bites an individual infected with dengue virus. The virus in the blood of the infected individual then infects the mosquito and travels from the mosquito's stomach to its salivary glands were the virus multiplies. The virus is then injected into another person when the mosquito injects anticoagulants that prevent blood clotting when the mosquito is feeding. The mosquito remains able to transmit dengue for its entire life. 3. Yellow fever virus is a potentially fatal viral infection that's transmitted by mosquitoes in tropical regions. It has both an urban cycle and a jungle cycle that relies on monkeys as carriers ('sylvatic cycle').Yellow fever virus belongs to the Flaviviridae family, other members of which cause dengue fever and Japanese encephalitis. 4. Japanese encephalitis virus previously known as Japanese B encephalitis to distinguish it from von Economo's A encephalitisis a disease caused by the mosquito-borne Japanese encephalitis virus. The Japanese encephalitis virus is a virus from the family Flaviviridae. Domestic pigs and wild birds are reservoirs of the virus; transmission to humans may cause severe symptoms. One of the most important vectors of this disease is the mosquito Culex tritaeniorhynchus. This disease is most prevalent in Southeast Asia and the Far East. 5 West Nile virus (WNV) is a virus of the family Flaviviridae. Part of the Japanese encephalitis (JE) antigenic complex of viruses, it is found in both tropical and temperate regions. It mainly infects birds, but is known to infect humans, horses, dogs, cats, bats, chipmunks, skunks, squirrels, and domestic rabbits. The main route of human infection is through the bite of an infected mosquito. Pathogenesis 1. St. Louis Encephalitis: transmitted by mosquito; bird reservoir; infects reticuloendothelial tissue, then viremia and dissemination to CNS 2. Dengue virus: transmitted by mosquito and vertically to neonate; primate and mosquito reservoir; infects reticuloendothelial tissue, then viremia (virus circulates in blood cells) with dissemination to many

tissues; inflammatory cytokines associated with pathogenesis and symptoms. 3. Yellow Fever virus: transmitted by mosquito; primate reservoir; pathogenesis similar to Dengue except hepatitis, nephritis and circulatory failure more prominent. 4. Japanese encephalitis virus : transmitted by mosquito; pigs and aquatic birds main reservoir 5. West Nile virus : transmitted by mosquito; crows and aquatic birds main reservoir Effects on human 1. St. Louis encephalitis virus causes fever & headache, meningitis, encephalitis 2. Dengue virus causes dengue fever (self-limited disease), Dengue hemorrhagic fever, Dengue shock syndrome 3. Yellow fever virus causes self-limited disease, hemorrhagic form 4. Japanese encephalitis virus causes asymptomatic infections or encephalitis 5. West Nile virus causes same as St. Louis encephalitis virus Infectious Cycle 1. Attachment- attachment of viral capsids/spikes and protein to specific receptor to host cell. 2. Penetration- force/endocytosis or membrane fussion. 3. Uncoating- capsid is being removed and dissolved through host cell enzymes leaving only the genetic material. 4. Biosynthesis- synthesis of viral messenger RNA/viral protein synthesis or assembly of viral protein and viral genom. 5. Release Incubation Period 1.St. Louis Encephalitis -incubation period is 21 days. 2. Dengue virus -3 to 15 days (usually lasting for 5-8 days) before the characteristics of dengue appear 3. Yellow fever virus -3 to 16 days of incubation period. 4. Japanese encephalitis virus -5 to 15 days of incubation period.

5. West Nile virus -5 to 15 days of incubation period. Laoratory Diagnosis 1.St.Louis Encephalitis Blood tests are critical to diagnosing SLE. Blood tests such as blood count and liver and kidney function tests may show mild abnormalities. Proteins, small amounts of red blood cells and white cells may also be seen in the urine during urine tests. Cerebrospinal fluid analysis. The cerebrospinal fluid (CSF) is the fluid that surrounds the brain, and it carries the virus. It is sampled by way of a lumbar puncture. The characteristic findings are an increased pressure and a slightly elevated protein count. Cerebrospinal fluid sampling may distinguish SLE from bacterial meningitis.

2.Dengue Virus

Complete blood cell count findings include the following: o Leukopenia, often with lymphopenia, is observed near the end of the febrile phase of illness. Lymphocytosis, with atypical lymphocytes, commonly develops before defervescence or shock. A recent systematic review found that patients with dengue had significantly lower total WBC, neutrophil, and platelet counts than patients with other febrile illnesses in dengue-endemic populations.27
o

A hematocrit level rise of greater than 20% is a sign of hemoconcentration and precedes shock. The hematocrit level should be monitored at least every 24 hours to facilitate early recognition of dengue hemorrhagic fever and every 3-4 hours in severe cases of dengue hemorrhagic fever or dengue shock syndrome. Thrombocytopenia has been demonstrated in up to 50% of dengue fever cases. Platelet counts of less than 100,000 cells/L are seen in dengue hemorrhagic fever or dengue shock syndrome and occur before defervescence and the onset of shock. The platelet count should be monitored at least every 24 hours to facilitate early recognition of dengue hemorrhagic fever.

Basic metabolic panel findings include the following:

o

Hyponatremia is the most common electrolyte abnormality in patients with dengue hemorrhagic fever or dengue shock syndrome.

Metabolic acidosis is observed in those with shock and must be corrected rapidly. Elevated BUN levels are observed in those with shock. Acute kidney injury is uncommon.28,29

Liver injury panel findings include the following:

o

Transaminase levels may be mildly elevated into the several thousands in patients with dengue hemorrhagic fever who have acute hepatitis. Low albumin levels are a sign of hemoconcentration.

Coagulation studies may help to guide therapy in patients with severe hemorrhagic manifestations. Findings are as follows:
o o o

Prothrombin time is prolonged. Activated partial thromboplastin time is prolonged. Low fibrinogen and elevated fibrin degradation product levels are signs of disseminated intravascular coagulation.

Typing and crossmatching of blood should be performed in cases of severe dengue hemorrhagic fever or dengue shock syndrome because blood products may be required. Serum specimens should be sent to the laboratory for serodiagnosis, PCR, and viral isolation. Because the signs and symptoms of dengue fever are nonspecific, attempting laboratory confirmation of dengue infection is important.
o

Serodiagnosis is made based on a rise in antibody titer in paired IgG or IgM specimens. Results vary depending on whether the infection is primary or secondary. The IgM capture enzyme-linked immunosorbent assay (MACELISA) has become the most widely used assay, although other tests, including complement fixation (CF), neutralization test (NT), hemagglutination inhibition (HI), and IgG ELISA are also used. A recent European study found that, if only a single serum sample is available, a single positive result on ELISA (PanBio IgM or IgG) was found to have a high rate of false positivity and should be confirmed using a second more specific diagnostic technique.30,31

In order to provide a more rapid reliable diagnosis, clinically available PCR studies are being developed.32,33

Cultures of blood, urine, CSF, and other body fluids should be performed as necessary to exclude or confirm other potential causes of the patient's condition.

3.Yellow Fever Virus Specific diagnosis depends on isolation of virus from blood, demonstration of viral antigen in serum by enzime-linked immunosorbent assay ( ELISA ) or of viral RNA by polimerase chain reaction ( PCR ) during the period of infection. Serologic diagnosis include IgM antibody-capture ELISA, hemagglutination inhibition (HI), complement fixation (CF) or neutralization (N) tests. Its important to consider that IgM, HI and N antibodies appear within 5 - 7 days and CF antibodies within 7 - 14 days after onset. Thus, paired acute and convalescent sera should be tested ( interval of 14 days ). Liver biopsy confirms the diagnosis by isolation of virus ( direct immunofluorescence or DNA hibridization ), but its absolutely contraindicated because of the bleeding diathesis; pathologic examination of the liver may suggest, but not assure a postmortem diagnosis. 4.Japanese Encephalitis Virus

A CBC count often shows nonspecific modest leukocytosis in the first week of illness. This may be followed by a relative leukopenia. A mild anemia may also be present. In one study, 15% of children with Japanese encephalitis had thrombocytopenia. Serum sodium levels may be depressed secondary to inappropriate antidiuretic hormone secretion. A study of Indian children during the Uttar Pradesh Japanese encephalitis outbreak in 2005 noted elevated liver function test results in a large number of patients (all had elevated aspartate aminotransferase [AST] levels; 47.2% had elevated alanine aminotransferase levels).2 Overall, the diagnosis of Japanese encephalitis may be supported by a capture immunoassay methodology demonstrating IgM antibody in the CSF. Alternatively the serum antibody level may be increased 4-fold. Isolation of Japanese encephalitis virus (JEV) from clinical specimens or even the identification of positive genetic viral sequences in tissue, blood, or CSF is diagnostic.

IgM antibody levels may be found even within 7 days of symptoms. Notably, JEV has been isolated up to even almost 4 months after clinical symptoms have begun. For laboratory worker safety, a biosafety level 3 is required for working with JEV. Serologic cross-reactivity among other viruses, specifically dengue and West Nile virus, may lead to confusion in the diagnostic evaluation of Japanese encephalitis (especially the tropical Asian regions).

5.West Nile Virus West Nile virus (WNV) testing for patients with encephalitis, meningitis, or other serious central nervous system infections can be obtained through local or state health departments. For WNV diagnosis, public health laboratories usually perform an IgM antibody capture enzyme-linked immunosorbent assay (MAC-ELISA). Using this assay, virus-specific IgM can be detected in nearly all cerebrospinal fluid (CSF) and serum specimens received from WNV-infected patients at the time of their clinical presentation. Because serum IgM antibody may persist for more than a year, physicians must determine whether the antibody is the result of a WNV infection in the previous year and unrelated to the current clinical presentation. The following procedures are recommended:

The most conclusive diagnostic method to identify persons with WNV infection of the central nervous system (CNS) is detecting WNV-specific IgM antibody in CSF using MAC-ELISA. This can be done with a CSF specimen obtained during initial clinical presentation. Because IgM antibody does not readily cross the blood-brain barrier, IgM antibody in CSF strongly suggests acute CNS infection If CSF is not obtained and serum samples are used to make the diagnosis, paired acute- and convalescent-phase serum samples should be acquired. The acute-phase specimen should be obtained during initial clinical presentation and the convalescent-phase specimen should be obtained 7-14 days later. Both samples should be tested with MAC-ELISA. If a convalescent-phase specimen cannot be obtained, the acute-phase specimen should be tested with MAC-ELISA. If the specimen is IgMnegative, then the illness is very unlikely to be an acute WNV infection. If the specimen is IgM-positive and the illness is clinically compatible, then it may be a recent WNV infection (presuming the test results for IgM antibody to St. Louis encephalitis (SLE) virus are significantly lower or negative; see below).

Ideally, MAC-ELISA testing should be performed, using both WNV and SLE virus. If the MAC-ELISA results for WNV and SLE are similar, it is necessary to use the plaque-reduction neutralization test (PRNT) to confirm either a WNV or SLE virus infection. Note: Patients who have been recently vaccinated against or recently infected with related flaviviruses (e.g., yellow fever, Japanese encephalitis, dengue) may have positive WNV MAC-ELISA results.

Medication 1.St.Louis Encephalitis No antiviral agent is available for the treatment of St. Louis encephalitis virus (SLEV) infection. A pilot study has shown that early use of interferon-alpha2b may decrease the severity of complications.No vaccine is available for pre-exposure protection. No vaccine is available for pre-exposure protection.

2.Dengue Fever Virus No specific antiviral medication currently is available to treat dengue infections. Single-dose methylprednisolone showed no mortality benefit in the treatment of dengue shock syndrome (dengue shock syndrome) in a prospective, randomized, double-blind, placebo-controlled trial. Acetaminophen (paracetamol) is recommended for treatment of pain and fever. Aspirin, other salicylates, and NSAIDs should be avoided. Analgesics/antipyretics The treatment of dengue fever is symptomatic and supportive in nature. Bedrest and mild analgesic-antipyretic therapy are often helpful in relieving lethargy, malaise, and fever associated with the disease. 3.Yellow Fever Virus Vaccines The live attenuated virus (17D) vaccine was created by serial passages of yellow fever virus through chick and mouse embryo cells. Dr. Max Theiler of the Rockefeller Institute developed this vaccine in 1937. Since 1945, more than 200,000,000 doses have been administered. The WHO, United Nations

Children's Fund (UNICEF), and the World Bank have recommended that yellow fever vaccine be added to the routine Expanded Program on Immunization in developing nations. However, poor financing remains a problem and a major reason for low vaccination rates among residents of endemic areas. In the United States, the yellow fever vaccine is available at designated state health departments and selected travel clinics. Yellow fever vaccine (YF-VAX) This vaccine should be administered to residents of and travelers to endemic areas. The seroconversion rate for adults and children receiving the vaccine is 99%. Protective antibodies form within 7-10 d, and protection lasts for at least 10 y. Vaccine is safe and effective in asymptomatic adult patients with HIV and CD4 counts >200/L. The vaccine appeared ineffective when administered to 1-year-old infants who were HIV positive (CD4 count >200/L). 4.Japanese Encephalitis Virus Osmotic diuretics Mannitol is recommended by some experts to help reduce intracranial pressure. Mannitol induces diuresis, which increases serum osmotic concentration. In the brain, this causes water to flow from brain cells into vascular space, thereby decreasing intracranial pressure. Mannitol (Osmitrol, Resectisol) Drug may be used to decrease intracranial pressure. May reduce subarachnoid space pressure by creating osmotic gradient between CSF in arachnoid space and plasma. Not for long-term use. Initially assess for adequate renal function by administering a test dose of 200 mg/kg IV over 3-5 min. It should produce a urine flow of at least 30-50 mL/h of urine over 2-3 h. In children, assess for adequate renal function by administering a test dose of 200 mg/kg IV over 3-5 min. It should produce a urine flow of at least 1 mL/h over 1-3 h. Vaccine Induction of antibody response to vaccine provides capability to neutralize live JEV.

Japanese encephalitis vaccine (Ixiaro, JE-VAX) Indicated for active immunization to prevent Japanese encephalitis virus (JEV) infection. Ixiaro: Inactivated vaccine prepared by propagating JEV strain SA14 -14-2 in Vero cells. Available in prefilled, single-dose syringes without preservatives or thimerosal. JE-VAX: Murine-derived inactivated vaccine derived from the Nakayama JEV strain. Available in a multidose vial. Contains thimerosal and gelatin. 5.West Nile Virus No specific drug treatment exists for West Nile encephalitis (WNE).

BUNYAVIRUS

-Spherical particles contain a single negative-sense RNA genome that is segmented. They have lipid-containing envelope and measure 90-100 nm. The nucleocapsids have helical syummetry and contain a major viral protein. The envelope has 2 glycoproteins in the lipid bilayer and surface projections (10nm) of glycopeptides clustered to form hollow cylinders. Several produce mosquito-borne encephalitides of humans and animals, others hemorraghic fevers. Some are transmitted by sandflies. Morphology Enveloped, spherical particles, 80-120nm diameter.

Nucleocapsid: Helical, 3 segments, 2 proteins - nucleocapsid ('N'), RNA polymerase ('L') Envelope: 2 glycoproteins, G1 and G2, spikes not obvious

Pathogenesis Varied, because they are a very large group of viruses, but generally: Insect bite results in transient viraemia; replication then occurs in target organs varies from one virus to another, as does severity (mild to severe). Infectious Cycle 1. Attachment- attachment of viral capsids/spikes and protein to specific receptor to host cell. 2. Penetration- force/endocytosis or membrane fussion.

3. Uncoating- capsid is being removed and dissolved through host cell enzymes leaving only the genetic material. 4. Biosynthesis- synthesis of viral messenger RNA/viral protein synthesis or assembly of viral protein and viral genom. 5. Release Laboratory Diagnosis 1. Virus Isolation - intracranial inoculation of suckling mice is thought to be the most sensitive system available for virus isolation. However, several sensitive cell culture systems are available such as vero. LLCMC2 and mosquito cells. Once isolated the virus can be types by neutralizing tests. 2. Rapid Diagnosis - antigen detection systems and the detection of specific IgM antibodies are becoming available as means of rapid diagnosis. 3. Serology - a wide variety of serological techniques are available such as HI, CFT, IFA, neutralization tests and ELISAs.

Major Strain of Bunyaviridae: 1.California Encephalitis California encephalitis is an arbovirus-induced, arthropod-borne encephalitis or encephalomeningitis. The virus is transmitted to humans through a mosquito bite. Pathogenesis California Encephalitis -After inoculation via a mosquito bite, the virus undergoes a local replication at the original skin site. A primary viremia occurs, with seeding of the reticuloendothelial system, mainly the liver, spleen, and lymph nodes. With continued virus replication, a secondary viremia occurs, with seeding of the CNS. The probability of CNS infection depends on the efficiency of viral replication at the extraneural sites and the degree of viremia. The virus invades the CNS through either the cerebral capillary endothelial cells or the choroid plexus. Rarely, the virus is isolated from brain tissue.

In a recent study involving mice, Bennett and colleagues (2008) reported that, even with systemic viremia, significant virus reproduction occurs in the nasal turbinates, and the virus may seed the CNS via the olfactory nerves.1 Antibodies against the G1 part of the virus neutralize the virus, block fusion, and inhibit hemagglutination. They are also important in virus clearance and recovery and in prevention of reinfection.

Effects on Human Malaise Rash Fever Myalgia Stiff neck Headache Photophobia Lethargy Coma Seizures Aphasia Anosmia

Infectious Cycle 1. Attachment- attachment of viral capsids/spikes and protein to specific receptor to host cell. 2. Penetration- force/endocytosis or membrane fussion. 3. Uncoating- capsid is being removed and dissolved through host cell enzymes leaving only the genetic material.

4. Biosynthesis- synthesis of viral messenger RNA/viral protein synthesis or assembly of viral protein and viral genom. 5. Release Incubation Period The incubation period is usually 3-7 days. Laboratory Diagnosis

According to the Centers for Disease Control and Prevention (CDC) guidelines for the diagnosis of arboviral encephalitis, febrile illness or mild aseptic meningitis or encephalitis (with onset during a period when the transmission of the virus is likely) occurs with one of the following: o A 4-fold increase in the antivirus antibody titer between the acute and the convalescent periods
o

Virus isolation from tissue, blood, or cerebrospinal fluid (CSF): Note that La Crosse virus has not been isolated from CSF. Specific immunoglobulin M (IgM) antibodies to the virus detected using enzyme-linked immunosorbent assay (ELISA) technique during the acute illness

Significant antibody titers include levels of more than 320 by hemagglutination inhibition, more than 128 by complement fixation, more than 256 by immunofluorescence, or more than 160 by plaque reduction neutralization test. CSF examination reveals the following:
o o o

Normal to mildly elevated pressure level Normal glucose level and normal to mildly elevated protein level Initially, a polymorphonuclear leukocytic pleocytosis followed by lymphocytic or monocytic leucocytosis is present.

Complete blood cell count is usually within the reference range or might show mild leucocytosis. Chemistries are usually within the reference range. Use of the polymerase chain reaction for the diagnosis of La Crosse encephalitis is still in the research stage.

Medication

No antiviral agent is available. No vaccine is available for preexposure protection.

Other Classification of Arbovirus: Reovirus- a few arboviruses are members of the genus orbivirus, including African horse sickness and Colorado tick fever. Some infect birds, small mammals, and ticks. Arenaviruses- pleomorphic particles contain a segmented single negative-sense RNA genome, are surrounded by an envelope, and measure 50-300 nm. They contain granules believed to be robosomes. Several hemorraghic fever viruses are member of this group. Most have rodent host in their natural cycle. Rhabdoviruses- several bullet-shaped arboviruses fall into this group.

RNA NON-ENVELOPED VIRUSES Picornavirus - derived from pico, which means small (typically, 18-30 nm). -A picornavirus is a virus belonging to the family Picornaviridae. Picornaviruses are non-enveloped, positive-stranded RNA viruses with an icosahedral capsid. The genome RNA is unusual because it has a protein on the 5' end that is used as a primer for transcription by RNA polymerase. The name is derived from pico meaning small, and RNA referring to the ribonucleic acid genome, so "picornavirus" literally means small RNA virus. Picornaviruses are separated into 12 distinct genera and include many important pathogens of humans and animals.[1] The diseases they cause are varied, ranging from acute "common-cold"-like illnesses, to poliomyelitis, to chronic infections in livestock. -includes 2 major groups: Enteroviruses and Rhinoviruses Morphology The virus has an IRES (Internal Ribosomal Entry Site) which distinguishes it from many other RNA viruses. The virus is naked with an icosahedral capsid. The triangulation number is 3, while the capsid has four unique proteins: VP1, 2, 3, and 4.

The capsid is one of the smallest of all viruses with a diameter of only 27-30nm. Translation and cleavage of viral polypeptides produces eleven distinct proteins.

Calicivirus- derived from the Latin word calyx meaning cup or goblet. This name is appropriate as many strains have visible cup-shaped depressions. -are a family of viruses, members of Class IV of the Baltimore scheme. They are positive-sense, single stranded RNA which is non-segmented. The caliciviruses have been found in a number of organisms such as humans, cattle, pigs, chickens, reptiles, dolphins and amphibians. The caliciviruses have a simple construction and are not enveloped. Morphology The capsid appears hexagonal/spherical and has icosahedral symmetry with a diameter of 35-39 nm. Reovirus- is derived from respiratory enteric orphan viruses.] The term "orphan virus" means that a virus that is not associated with any known disease. -a family of viruses that can affect the gastrointestinal system (such as Rotavirus) and respiratory tract. Viruses in the family Reoviridae have genomes consisting of segmented, double-stranded RNA (dsRNA). Morphology Icosahedral capsid (T-13) composed of an outer and inner protein shell. Genomes of viruses in Reoviridae contain 10-12 segments which are grouped into three categories corresponding to their size: L (large), M (medium) and S (small). Segments range from ~ 3.9 kbp 1kbp and each segment encodes 1-3 proteins.

Virus a.dna enveloped viruses a1.herpes simplex virus 1&2 a.2 varicella-zoster virus a3.cytomegalovirus a4.epstein-barr virus a5.hepatitis b virus b.dna on-enveloped b1.adenovirus

c.rna enveloped virus c1.orthomyxovirus -influenza virus c.2 paramyxovirus c2.1.parainfluenza virus c2.2.mumps virus c2.3.measles virus. C2.4. respiratory syncitial virus