Molecularbiological diagnosis of herpes virus infection of a juvenile Russian tortoise (Agrionemys horsfieldii) with skin and lung lesions

Herpesvirus infections are significant in the care of turtles and tortoises. Clinical signs range from unspecific symptoms, due to the variety of organ manifestations, to the "classical" picture of rhinitis-stomatitis. The presented case study showed the typical disease only with respect to clinical symptoms following hibernation, but lacks stomatitis, erosions or plaques in the oral mucosa. On the other hand, skin lesions on the extremeties, causative with herpesvirus infection, could be diagnosed. In this case study, various symptoms, sampling procedures and diagnostics using two different PCR methods are presented. Following hibernation, samples from a Russian tortoise (Agrionemys horsfieldii) were taken ante mortem and post mortem and screened with respect to virology, pathology, bacteriology and parasitology. DNA-fragments specific for tortoise herpesvirus were detected in various organs and body liquids. Furthermore basophilic intranuclear inclusion bodies were found. The bacteriological examination showed a high level of Pasteurella testunis in the lungs. By parasitological examination nematodes (Oxyridae) were diagnosed. A potential prophylaxis tortoise herpesvirus is discussed.     

Pathologic changes of experimental simian varicella (Delta herpesvirus) infection in African green monkeys (Cercopithecus aethiops)

Gross and microscopic lesions of experimental simian varicella (Delta herpesvirus) infection in African green monkeys (Cercopithecus aethiops) are described. In stratified squamous epithelium, such as the epidermis, tongue, and esophagus, focal areas of epidermal hyperplasia, ballooning degeneration, and blood-filled vesicle formation with ulceration were apparent. Most visceral organs were involved, and the changes included necrosis, hemorrhage, and characteristic intranuclear eosinophilic inclusions in a variety of cells. A generalized vascular involvement was present with intranuclear inclusions in the endothelial and smooth muscle cells in various organs. The nervous system was normal on gross and microscopic examination. Virus titer and serum transferase values were correlated with the clinical signs of infection.     

BRS virus, PI3 virus and BHV1 infections of young stock on self-contained dairy farms: epidemiological and clinical findings

The role of bovine respiratory syncytial virus, parainfluenza type 3 virus and bovine herpesvirus 1 as disease agents in 28 groups of young cattle on 19 dairy farms which raised their own replacements was investigated. Bovine respiratory syncytial virus infections occurred in 27, parainfluenza type 3 virus infections in all and bovine herpesvirus 1 infections in three of the 28 groups. Some infections were accompanied by clinical signs while others were entirely subclinical. Clinical respiratory disease was observed on 25 occasions in 20 of the groups. Respiratory disease was associated with a bovine respiratory syncytial virus infection on 15 occasions with parainfluenza type 3 virus infection in four cases and with bovine herpesvirus 1 infection in two cases. In four cases there was no association between the respiratory disease and any of the four virus infections. Bovine respiratory syncytial virus infections caused more serious respiratory problems than parainfluenza type 3 virus infections.     

Feeding Cyprinus carpio with infectious materials mediates cyprinid herpesvirus 3 entry through infection of pharyngeal periodontal mucosa

ABSTRACT: Cyprinid herpesvirus 3 (CyHV-3), also known as Koi herpesvirus, is the etiological agent of a mortal disease in common and koi carp. Recently, we investigated the entry of CyHV-3 in carp using bioluminescence imaging and a CyHV-3 recombinant strain expressing luciferase (LUC). We demonstrated that the skin is the major portal of entry after inoculation of carp by immersion in water containing CyHV-3. While this model of infection mimics some natural conditions in which infection takes place, other epidemiological conditions could favour entry of virus through the digestive tract. Here, we investigated whether ingestion of infectious materials mediates CyHV-3 entry through the digestive tract. Carp were fed with materials contaminated with the CyHV-3 LUC recombinant (oral contamination) or immersed in water containing the virus (contamination by immersion). Bioluminescence imaging analyses performed at different times post-infection led to the following observations: (i) the pharyngeal periodontal mucosa is the major portal of entry after oral contamination, while the skin is the major portal of entry after contamination by immersion. (ii) Both modes of inoculation led to the spreading of the infection to the various organs tested. However, the timing and the sequence in which some of the organs turned positive were different between the two modes of inoculation. Finally, we compared the disease induced by the two inoculation modes. They led to comparable clinical signs and mortality rate. The results of the present study suggest that, based on epidemiological conditions, CyHV-3 can enter carp either by skin or periodontal pharyngeal mucosal infection.     

Infectious bovine rhinotracheitis: a review and update.

Infectious bovine rhinotracheitis (IBR), a herpesvirus infection of cattle, has diverse clinical manifestations. Known mainly as a respiratory tract disease characterized by tracheitis, rhinitis, and fever, IBR plays a prominent role among causes of undifferentiated bovine respiratory disease and abortion. It also causes conjunctivitis, infectious pustular vulvovaginitis, balanoposthitis, and rarely, encephalitis. The virus is readily transmitted and has worldwide distribution. Some cattle develop a latent infection, which can be reactivated. The disease can be diagnosed by clinical signs and lesions and by a variety of virologic and immunologic techniques. Control of IBR is based largely on vaccination. A review is timely because efficient utilization of the wide assortment of available vaccines requires knowledge of recent developments in the diagnosis, immunology, and epidemiology of the disease.     

Two herpesviruses associated with disease in wild Atlantic loggerhead sea turtles (Caretta caretta)

Herpesviruses are associated with lung-eye-trachea disease and gray patch disease in maricultured green turtles (Chelonia mydas) and with fibropapillomatosis in wild sea turtles of several species. With the exception fibropapillomatosis, no other diseases of wild sea turtles of any species have been associated with herpesviral infection. In the present study, six necropsied Atlantic loggerhead sea turtles (Caretta caretta) had gross and histological evidence of viral infection, including oral, respiratory, cutaneous, and genital lesions characterized by necrosis, ulceration, syncytial cell formation, and intranuclear inclusion bodies. Nested polymerase chain reaction targeting a conserved region of the herpesvirus DNA-dependent-DNA polymerase gene yielded two unique herpesviral sequences referred to as loggerhead genital-respiratory herpesvirus and loggerhead orocutaneous herpesvirus. Phylogenetic analyses indicate that these viruses are related to and are monophyletic with other chelonian herpesviruses within the subfamily alpha-herpesvirinae. We propose the genus Chelonivirus for this monophyletic group of chelonian herpesviruses.     

Outbreak of canine distemper in domestic ferrets (Mustela putorius furo)

In 2006 an outbreak of canine distemper affected 14 young domestic ferrets in Barcelona, Spain. Their clinical signs included a reduced appetite, lethargy, dyspnoea, coughing, sneezing, mucopurulent ocular and nasal discharges, facial and perineal dermatitis, diarrhoea, splenomegaly and fever. Late in the course of the disease, general desquamation and pruritus, and hyperkeratotic/crusting dermatitis of the lips, eyes, nose, footpads, and perineal area were observed. None of the ferrets developed neurological signs. Non-regenerative anaemia and high serum concentrations of alpha- and beta-globulins were the most common laboratory findings. Most of the animals died or were euthanased because of respiratory complications. Postmortem there were no signs of lung collapse. Distemper was diagnosed by direct immunofluorescence of conjunctival swabs or pcr of several organs, and histology revealed the characteristic eosinophilic intracytoplasmic and intranuclear inclusion bodies of canine distemper virus in several organs. The minimum incubation periods calculated for six of the ferrets were 11 to 56 days, and in 13 of the ferrets the signs of disease lasted 14 to 34 days. Inclusion bodies compatible with infection by herpesvirus were found in the lungs of one of the ferrets.     

PATHOGENICITY OF A STRAIN OF FELINE CALICIVIRUS FOR DOMESTIC KITTENS

A strain of feline calicivirus, isolated from a cheetah exhibiting ulcerative glossitis and conjunctivitis, was administered by aerosol to 4 domestic cats and by contact to a fifth cat. Despite the limited number of animals available for the experiment, the pathogenicity of the virus strain for domestic cats was established. In aerosol-infected animals, clinical signs were referable to infection of both upper and lower respiratory tracts. The virus produced an interstitial pneumonia which, early in infection, was uncomplicated by secondary bronchopneumonia. The in-contact cat exhibited clinical signs referable to infection of the oral cavity only and no lesions were noted in the lower respiratory tract at autopsy. Ulcerative glossitis was a feature of the disease in aerosol-infected and in-contact cats. The virus was isolated from the pharynx of all cats for up to 21 days after infection and from the tonsils at autopsy. The tonsils were considered to be a probable site of multiplication of virus in "carrier" cats.     

Identification of a novel herpesvirus from a California desert tortoise (Gopherus agassizii)

Herpesviruses are significant pathogens of tortoises, causing upper respiratory tract disease and necrotizing stomatitis, with infections often associated with high mortality rates. Herpesvirus infection in a captive California desert tortoise (Gopherus agassizii) was detected by light microscopic observation of intranuclear inclusion bodies in various tissues followed by transmission electron microscopic observation of herpesvirus-like particles, and amplification of herpesvirus nucleic acid sequences using polymerase chain reaction. Using an indirect enzyme linked immunosorbent assay, anti-tortoise herpesvirus antibodies were detected one month after initial onset of clinical signs. This novel herpesvirus is distinct from the previously described tortoise herpesvirus (tortoise herpesvirus-1, THV-1) sharing 83% sequence identity of 60 amino acids of a portion of the DNA polymerase gene and 79% sequence identity across 120 amino acids of a portion of the ribonucleotide reductase gene. Similar to THV-1, this novel herpesvirus, tortoise herpesvirus-2 (THV-2), also clusters with the alphaherpesviruses.     

Equine herpesvirus 1‐associated ulcerative dermatitis in a horse

This case report describes the clinical and histopathological findings of an infection caused by equine herpesvirus‐1 (EHV‐1) in a horse showing respiratory signs and a papular, crusted and ulcerative dermatitis involving mucosae. This diagnosis was supported by real‐time PCR positive for EHV‐1 on nasal swabs and tissues.     

Development of lesions and tissue distribution of parasite in lambs orally infected with sporulated oocysts of Toxoplasma gondii

The host-pathogen interaction is as a key feature during the formation of tissue cysts of Toxoplasma gondii within intermediate hosts. In this study, we investigated whether oral infection of lambs with T. gondii oocysts may be used as an experimental model in sheep to study this interaction, with the main objective being to detect the presence and distribution of lesions and parasite within different organs at different time points after oral infection. Lambs were infected with 5 × 10(3) and 5 × 10(5) sporulated T. gondii oocysts and culled at 2, 3, 5 and 6 weeks post-infection (WPI). During the infection, rectal temperature of the animals and serological antibodies against T. gondii were monitored. The presence of inflammatory lesions and parasite were evaluated through histological and immunohistochemical methods at different organs (brain, liver, lung, heart and lymph nodes). The lambs showed no clinical signs other than fever, and lesions appeared mainly in the brain, characterized by glial foci and perivascular cuffs, and in the heart, denoted by foci of interstitial myositis. Tissue cysts and tachyzoite-like structures were observed at all time points studied in the brain, where together with the glial foci they appeared mainly in the cerebral cortex of the forebrain and in the midbrain, but also in the heart, lung and lymph nodes. This study shows that oral infection with sporulated oocysts in lambs may provide a model for investigating the host-parasite interaction in situ during the development of tissue cysts.     

Chemoprophylaxis of cryptosporidiosis in chickens, using halofuginone, salinomycin, lasalocid, or monensin

Chemoprophylaxis of Cryptosporidium baileyi infections was attempted by feeding 4 groups of chicks diets containing 3 mg of halofuginone/kg of feed, 60 mg of salinomycin/kg, 75 mg of lasalocid/kg, or 110 mg of monensin/kg. Rations were fed 5 days before oral or intratracheal inoculation with oocysts and were continued for 20 days. None of the drugs prevented C baileyi infections. Clinical signs of respiratory tract disease and gross lesions of airsacculitis were observed in intratracheally inoculated birds in all treatment groups and nonmedicated controls. Orally inoculated birds did not develop clinical signs of infection. Pathogenic bacteria were not isolated from the respiratory tract systems of any chicks. Halofuginone delayed the establishment of infections of the bursa of Fabricius and cloaca, but not of the trachea.     

Comparisons between two isolates of stomatitis papulosa virus

The cytopathology and length of latency in single-step growth curves of two isolates of stomatitis papulosa virus are compared in this report. Isolate 721 was obtained from a calf with oral ulcers and isolate 8665 was obtained from a calf with respiratory disease and oral ulcers. In single-step growth curves, the latency period of isolate 721 was 8 h while that of isolate 8665 was 6 h. The cytopathic effect produced by isolate 721 in bovine lung cells was characterized by enlargement of the cell, cell-to-cell adherence and large intracytoplasmic accumulations of viral inclusion material. Isolate 8665 caused rapid cell degeneration and detachment, with small accumulation of viral inclusion material. Neither of the two strains grew in bovine alveolar macrophage cultures or in the respiratory epithelium of fetal bovine tracheal explants. Intragingival inoculation of these isolates in cattle resulted in oral lesions without clinical signs of respiratory of systemic involvement. Virus was recovered from the oral lesions and from nasal secretions for as long as 10 days. Inoculation of dexamethasone-treated cattle resulted in a similar clinical condition although virus was recovered for 20 days from oral lesions and nasal secretions. Seroconversions from negative to 1 : 2560 were detected in inoculated cattle by indirect immunofluorescence.     

Characterization of protection from systemic infection and disease by use of a modified-live noncytopathic bovine viral diarrhea virus type 1 vaccine in experimentally infected calves

OBJECTIVE: To evaluate protection against systemic infection and clinical disease provided by use of a modified-live noncytopathic bovine viral diarrhea virus (BVDV) type 1 vaccine in calves challenged with NY-1 BVDV. ANIMALS: 10 calves, 5 to 7 months of age. PROCEDURES: Calves were allocated (n = 5/group) to be nonvaccinated or vaccinated SC on day 0 with BVDV type 1 (WRL strain). Calves in both groups were challenged intranasally with NY-1 BVDV on day 21. Calves' rectal temperatures and clinical signs of disease were recorded daily, total and differential WBC and platelet counts were performed, and serum neutralizing antibody titers against NY-1 BVDV were determined. Histologic examinations and immunohistochemical analyses to detect gross lesions and distribution of viral antigens, respectively, were performed. RESULTS: After challenge exposure to NY-1 BVDV, nonvaccinated calves developed high rectal temperatures, increased respiratory rates, viremia, leukopenia, lymphopenia, and infection of the thymus. Vaccinated calves did not develop high rectal temperatures or clinical signs of respiratory tract disease. Vaccinated calves appeared to be protected against systemic replication of virus in that they did not develop leukopenia, lymphopenia, viremia, or infection of target organs, and infectious virus was not detected in peripheral blood mononuclear cells or the thymus. CONCLUSIONS AND CLINICAL RELEVANCE: The modified-live BVDV vaccine protected calves against systemic infection and disease after experimental challenge exposure with NY-1 BVDV. The vaccine protected calves against infection and viremia and prevented infection of target lymphoid cells.     

Comparative pathological studies on domestic geese (Anser anser domestica) and Muscovy ducks (Cairina moschata) experimentally infected with parvovirus strains of goose and Muscovy duck origin

Parvovirus infection of Muscovy ducks caused by a genetically and antigenically distinct virus has been reported from Germany, France, Israel, Hungary, some Asian countries and the USA. The pathological changes include those of degenerative skeletal muscle myopathy and myocarditis, hepatitis, sciatic neuritis and polioencephalomyelitis. In the study presented here, day-old and 3-week-old goslings and Muscovy ducks were infected experimentally with three different parvovirus strains (isolates of D-216/4 from the classical form of Derzsy's disease, D-190/3 from the enteric form of Derzsy's disease, and strain FM from the parvovirus disease of Muscovy ducks). All three parvovirus strains caused severe disease in both day-old and 3-week-old Muscovy ducks but in the goslings only the two strains of goose origin (D-216/4 and D-190/3) caused disease with high (90-100%) mortality when infection was performed at day old. Strain FM (of Muscovy duck origin) did not cause any clinical signs or pathological lesions in the goslings. In the day-old goslings and Muscovy ducks the principal pathological lesions were severe enteritis with necrosis of the epithelial cells (enterocytes) of the mucous membrane and the crypts of Lieberkühn, and the formation of intranuclear inclusion bodies. Other prominent lesions included hepatitis and atrophy (lymphocyte depletion) of the lymphoid organs (bursa of Fabricius, thymus, spleen). In goslings infected with the strain originating from the classical form of Derzsy's disease mild myocarditis was also detected. After infection at three weeks of age, growth retardation, feathering disorders, myocardial lesions (degeneration of cardiac muscle cells, lympho-histiocytic infiltration) and hepatitis were the most prominent lesions in both geese and Muscovy ducks. In addition to the lesions observed in the geese, muscle fibre degeneration, mild sciatic neuritis and polioencephalomyelitis were also observed in the Muscovy ducks infected with any of the three parvovirus strains.     

Equine Multinodular Pulmonary Fibrosis in association with asinine herpesvirus type 5 and equine herpesvirus type 5: a case report

A standardbred gelding with a history of 10 days pyrexia and lethargy was referred to the Equine Hospital at the Swedish University of Agricultural Sciences in Uppsala, Sweden.The horse had tachypnea with increased respiratory effort and was in thin body condition. Laboratory findings included leukocytosis, hyperfibrinogenemia and hypoxemia. Thoracic radiographs showed signs of pneumonia with a multifocal nodular pattern, which in combination with lung biopsy findings indicated Equine Multinodular Pulmonary Fibrosis (EMPF). EMPF is a recently described disease in adult horses with clinical signs of fever, weight loss and respiratory problems. The pathological findings include loss of functional pulmonary parenchyma due to extensive nodular interstitial fibrosis which has been related to infection with the equine herpesvirus type 5 (EHV-5). In this case, lung biopsy and tracheal wash samples tested positive for both asinine herpesvirus type 5 (AHV-5) and EHV-5 using PCR assays. The horse failed to respond to treatment and was euthanized for humane reasons. Postmortem examination confirmed the diagnosis of EMPF. This case suggests that not only EHV-5 alone should be considered in association with the development of this disease.     

Clinical, serological and molecular investigations of EHV-1 and EHV-4 in 15 unweaned thoroughbred foals

Fifteen unweaned thoroughbred foals, born on a stud farm to vaccinated mares, were clinically monitored during their first six months of life and repeatedly tested for equine herpesvirus type 1 (EHV-1) and equine herpesvirus type 4 (EHV-4). Nasopharyngeal swabs and blood samples were collected and screened respectively by PCR and seroneutralisation to detect the presence of the virus, explore its role as a possible cause of respiratory disease, and to assess the efficiency of the pcr for the diagnosis of this disease. The foals were divided into three groups on the basis of their clinical signs and whether they had seroconverted to EHV-1 and/or EHV-4: first, foals with no clinical signs of disease that had not seroconverted; secondly, foals with clinical signs that had seroconverted, and thirdly, foals with clinical signs that had not seroconverted. The results indicated that the viruses circulated on the stud farm despite stringent vaccination regimens against them, and confirmed their association with respiratory disease. The absence of significantly different pcr results among the three groups of foals showed that the pcr was effective in confirming the circulation of the viruses on the premises without being particularly helpful as a diagnostic tool.     

Experimental respiratory infection of goats with caprine herpesvirus and Pasteurella haemolytica

Groups of six male goats were inoculated intratracheally and intranasally with either caprine herpesvirus followed 6 days later by Pasteurella haemolytica, canine herpesvirus alone or P. haemolytica alone. Pneumonic lesions were observed in five of the six goats inoculated with caprine herpesvirus followed by P. haemolytica and in three of the six goats inoculated with P. haemolytica alone, but were not observed in goats inoculated with caprine herpesvirus alone or in non-infected controls. Pasteurella haemolytica was isolated from seven of eight lungs with pneumonia, but only from one of sixteen lungs without pneumonia. The lesions ranged from fatal acute exudative necrotising pneumonia to predominantly proliferative pneumonia. Half of the caprine herpesvirus-inoculated goats developed a clinical catarrhal rhinitis five days post-inoculation and the only virus-specific histopathological lesion was a mild tracheitis. Canine herpesvirus was recovered from the nasal swabs of all caprine herpesvirus- inoculated goats developed a clinical catarrhal rhinitis five days post-inoculation and the only virus-specific histopathological lesion was a mild tracheitis. Canine herpesvirus was recovered from the nasal swabs of all canine herpesvirus-inoculated goats and from the lungs of three goats inoculated with caprine herpesvirus alone. The experimental inoculations demonstrated that P. haemolytica alone can produce pneumonia in goats. In addition, the study showed that caprine herpesvirus readily proliferates in the upper respiratory tract and lungs of goats but the role of caprine herpesvirus in the aetiology of pneumonia remains uncertain.