Catarrhal Fever Research Articles

Using ELISA to detect pathogen antibodies in wild mammal carcasses: a systematic literature review

Abstract Carcasses of wild mammals provide convenient samples for wildlife disease investigators. Specifically, antibodies can persist in the blood and tissues of animals up to a few days after death and can provide information on exposure to specific pathogens, including microorganisms with zoonotic implications, such as Toxoplasma gondii, Trichinella spp., Brucella spp., and Mycobacterium spp. Although screening samples from decaying wildlife for antibodies is not a new method, an overview of the literature with clear recommendations on best practices is lacking. Here, we present results from a systematic review of 115 studies using enzyme‐linked immunosorbent assay (ELISA) to detect pathogen antibodies in wild mammals across 298 individual research projects. We discuss the range of mammalian hosts and the pathogens most represented in the literature and suggest improvements to consider when using samples from decaying animals for seroinvestigation. Our results indicate that the most examined mammals were artiodactyl ungulates and that antibodies against the bacteria Brucella spp. are those most examined to date, followed by the protozoan Toxoplasma gondii, malignant catarrhal fever virus, Schmallenberg orthobunyavirus, and the coccidian Neospora caninum. The effect of haemolysis in serum samples on ELISA results was considered in only 3.7% of the research projects we reviewed, sample size decisions were not outlined in 83% of research projects, and half of the projects did not examine seroprevalence in different age classes. Nevertheless, these details are important for replicability and accurate interpretation of results. We highlight the benefits and limitations of detecting antibodies against pathogens in carcasses from wild mammalian groups. Wild mammals are reservoirs for a wide range of pathogens including those with zoonotic potential and investigating their carcasses can help wildlife investigators to better understand the epidemiology of many diseases with implications for wildlife, livestock, and human health.

Molecular detection and phylogenetic analysis of ovine herpesvirus-2 in subclinical infections of cattle and sheep.

Ovine herpesvirus-2 (OvHV-2) is the causative agent of malignant catarrhal fever (MCF), a serious and often fatal disease that affects cattle and other ruminants. This study aimed to investigate the molecular epidemiology and genetic diversity of OvHV-2 strains circulating in sheep and cattle populations in the Jammu and Kashmir region of India. Screening of 150 sheep and 57 cattle blood samples revealed the presence of the OvHV-2 polymerase (pol) gene in 8.6% of sheep, 10% of apparently healthy cattle, and 29.7% of cattle exhibiting MCF-like symptoms. The full-length glycoprotein B (gB) gene (2800bp) and an 875bp internal fragment were successfully amplified, cloned, and sequenced from pol-positive samples. Comparative sequence analysis of the deduced gB amino acid sequences identified seven substitutions at positions 278, 341, 390, 440, 468, 539, and 566 compared to reference strains. Phylogenetic analysis based on the gB nucleotide sequences clustered the OvHV-2 strains from this study within the Indian clade, distinct from strains reported in the UK and US. These findings provide insights into the genetic diversity of OvHV-2 strains circulating in Jammu and Kashmir, with the identified mutations potentially influencing virus-host interactions. Further investigations into the functional implications of these mutations are warranted to understand their role in viral pathogenesis and tropism.

Transmission patterns of malignant catarrhal fever in sheep and cattle in Karnataka, India.

Malignant catarrhal fever (MCF) presents a sporadic yet significant threat to livestock and wildlife. A comprehensive investigation in Karnataka, India into the prevalence and transmission patterns of sheep-associated MCF (SA-MCF) was conducted. A total of 507 sheep peripheral blood leukocyte samples from 13 districts along with 27 cows and 10 buffalo samples from various regions in Karnataka were tested for SA-MCF infection i.e. Ovine gammaherpesvirus 2 (OvHV-2) using heminested PCR. Furthermore, serum samples collected from 73 cows and 15 buffalo suspected of MCF were tested using a commercially available ELISA kit. Additionally, histopathological examinations of affected tissues and phylogenetic analysis of viral tegument protein sequences were conducted. Our findings indicated a 20.11%, 33.33% and 20% positivity for OvHV-2 in sheep, cows and buffalo respectively by PCR. Statistical analysis revealed a significant association between the age of sheep and the detection of OvHV-2. Seven cows and one buffalo serum samples tested positive for ELISA. Clinical findings in bovids were consistent with typical MCF signs, and histopathological results revealed multi-organ involvement characterised by necrotising vasculitis and lymphoid hyperplasia. The nucleotide pairwise identity matrix revealed 99.5% identity between the sequences obtained in the study with sequences from other states. The phylogenetic analysis of partial tegument protein sequences from bovid and sheep samples suggested a close genetic relationship between the local OvHV-2 strains and those from various global regions. Crucially, this study underscores the widespread presence of SA-MCF in Karnataka, with significant implications for both livestock management and wildlife conservation.

Transplacental Infections Associated with Macavirus in Aborted Bovine Fetuses.

The Macavirus genus, Gammaherpesvirinae subfamily, Herpesviridae family, contains ovine gammaherpesvirus 2 (OvGHV2), the cause of sheep-associated malignant catarrhal fever (SA-MCF). Members of the Macavirus genus associated with the development of malignant catarrhal fever (MCF) in their respective hosts share the 15A antigenic epitope, are conserved within the DNA polymerase gene and are collectively referred to as the malignant catarrhal fever virus (MCFV) complex. The ability of MCFV and/or OvGHV2 to produce abortions in ruminants is currently unknown, with little documentation of infections by these agents in bovine fetuses. This report presents the findings observed due to the detection of OvGHV2 DNA and MCFV tissue antigens in aborted bovine fetuses from southern Brazil. Four aborted bovine fetuses from three farms, located in a geographical region of Paraná State with elevated immunohistochemical (IHC) prevalence of MCFV tissue antigens, with gestational ages varying between 78 to 208 days were investigated. Significant gross and histopathological alterations were not observed in any of these fetuses. An IHC assay using the 15A-monoclonal antibody (15A-MAb), which is based on the 15A antigenic epitope of Macavirus, identified MCFV tissue antigens in multiple organs from two fetuses (#1 and #4); however, positive immunoreactivity to the 15A-MAb IHC assay was not detected in Fetus #2 and #3. Molecular testing amplified OvGHV2 DNA only from the myocardium and lungs of Fetus #1 that had positive intracytoplasmic immunoreactivity to the 15A-MAb IHC assay in these tissues. Furthermore, infections by Leptospira spp. were confirmed by molecular assays in fetuses #1, #3, and #4, while PCR detected Neospora caninum in the myocardium of Fetus #2. Additionally, molecular assays to identify well-known fetopathy agents of cattle, including bovine viral diarrhea virus, bovine alphaherpesvirus 1, Histophilus somni, and Listeria monocytogenes, did not amplify the nucleic acids of these pathogens. PCR assays to identify bovine gammaherpesvirus 6 (BoGHV6), another Macavirus known to infect cattle in Brazil, were unsuccessful. These findings confirmed that the 15A-MAb IHC assay can be efficiently used to detect MCFV antigens in organs of aborted bovine fetuses. The identification of MCFV antigens with the simultaneous detection of OvGHV2 DNA confirmed that Fetus #1 was infected by OvGHV2 and added to the few descriptions of this infection in aborted fetuses of ruminants worldwide. Moreover, the IHC detection of MCFV in multiple organs of Fetus #4, without the molecular detection of OvGHV2 or BoGHV6, may suggest that this fetus was infected by a Macavirus that was not previously diagnosed in cattle herds from Brazil. These findings strongly suggest that OvGHV2 and MCFV can produce transplacental infections in cattle.

Unraveling clonal CD8 T cell expansion and identification of essential factors in γ-herpesvirus-induced lymphomagenesis

Alcelaphine gammaherpesvirus 1 (AlHV-1) asymptomatically persists in its natural host, the wildebeest. However, cross-species transmission to cattle results in the induction of an acute and lethal peripheral T cell lymphoma-like disease (PTCL), named malignant catarrhal fever (MCF). Our previous findings demonstrated an essential role for viral genome maintenance in infected CD8+ T lymphocytes but the exact mechanism(s) leading to lymphoproliferation and MCF remained unknown. To decipher how AlHV-1 dysregulates T lymphocytes, we first examined the global phenotypic changes in circulating CD8+ T cells after experimental infection of calves. T cell receptor repertoire together with transcriptomics and epigenomics analyses demonstrated an oligoclonal expansion of infected CD8+ T cells displaying effector and exhaustion gene signatures, including GZMA, GNLY, PD-1, and TOX2 expression. Then, among viral genes expressed in infected CD8+ T cells, we uncovered A10 that encodes a transmembrane signaling protein displaying multiple tyrosine residues, with predicted ITAM and SH3 motifs. Impaired A10 expression did not affect AlHV-1 replication in vitro but rendered AlHV-1 unable to induce MCF. Furthermore, A10 was phosphorylated in T lymphocytes in vitro and affected T cell signaling. Finally, while AlHV-1 mutants expressing mutated forms of A10 devoid of ITAM or SH3 motifs (or both) were able to induce MCF, a recombinant virus expressing a mutated form of A10 unable to phosphorylate its tyrosine residues resulted in the lack of MCF and protected against a wild-type virus challenge. Thus, we could characterize the nature of this γ-herpesvirus-induced PTCL-like disease and identify an essential mechanism explaining its development.

Subclinical Ovine Gammaherpesvirus 2-Related Infections in Free-Ranging Wild Boars (Sus scrofa) from Southern Brazil.

Ovine gammaherpesvirus 2 (OvGHV2), is a Macavirus and the cause of sheep-associated malignant catarrhal fever (SA-MCF), in which sheep are the asymptomatic reservoir hosts. Susceptible mammalian populations infected by OvGHV2 may develop clinical SA-MCF or subclinical infections. All members of the Macavirus genus known to be associated with MCF are collectively referred to as the MCF virus (MCFV) complex. This report describes the occurrence of subclinical OvGHV2-related infections in free-ranging wild boars (Sus scrofa) from southern Brazil. Specific body organs (n = 14) and biological samples (nasal and oral swabs; n = 17) were collected from 24 asymptomatic wild boars from a conservation unit located within the Central-eastern mesoregion of Paraná State. Organs were processed to observe histopathological patterns suggestive of diseases of domestic animals; only pulmonary samples were used in an immunohistochemical assay designed to detect MCFV tissue antigens. Furthermore, all samples were submitted to molecular assays designed to detect the OvGHV2 tegument protein gene. Viral-induced pneumonia was diagnosed in two wild boars; one of these contained OvGHV2 DNA, with MCFV antigens identified in the other. Additionally, MCFV tissue antigens were detected within pulmonary epithelial cells of the lungs with and without pulmonary disease. Collectively, OvGHV2 was detected in 37.5% (9/24) of all wild boars, with detection occurring in the organs of 57.1% (8/14) wild boars and the oral cavity of one animal. These results demonstrated that these wild boars were subclinically infected by OvGHV2, and that infection produced typical pulmonary alterations. In addition, the detection of OvGHV2 within the oral cavity of one wild boar may suggest that this animal may be a potential disseminator of this pathogen to susceptible animal populations, including livestock and wildlife, acting as a possible bridge host for OvGHV2. Furthermore, infection by OvGHV2 probably occurred due to incidental contact with asymptomatic sheep maintained within the surrounding rural areas and not within the conservation units.

First reported case in an Irish flock of MCF- like systemic necrotizing vasculitis in sheep associated with ovine herpesvirus 2

BackgroundOvine gammaherpesvirus 2 (OvHV-2) is the causative agent of sheep associated malignant catarrhal fever (MCF). As sheep are the adapted host for OvHV-2, it is generally presumed that infection is not associated with disease in this species. However, a recent case review combined in-situ hybridisation, PCR and histopathology and correlated the viral distribution with systemic necrotizing vasculitis and concluded OvHV-2 was the likely agent responsible for sporadic, MCF-like vascular disease in sheep.Case presentationUsing similar methods this case study reports on the findings of the first reported cases in an Irish Flock of MCF- like systemic necrotizing vasculitis in sheep associated with OvHV-2. Sheep A, a 16-month-old Texel-cross hogget displayed signs of ill- thrift, Sheep B, a nine-month-old Belclare-cross lamb, was found dead having displayed no obvious symptoms. Both cases occurred on the same farm, however the animals were not related. Lymphohistiocytic vasculitis of various tissues was the predominant histopathological finding in both animals.ConclusionBy combining histopathology, PCR and in-situ hybridisation results, MCF- like systemic necrotizing vasculitis associated with OvHV-2 has been diagnosed for the first time in an Irish flock.

Quantification of ovine gammaherpesvirus 2 in clinical cases of cattle with sheep-associated malignant catarrhal fever.

Ovine gammaherpesvirus 2 (OvGHV2) produces sheep-associated malignant catarrhal fever (SA-MCF), a frequently lethal, lymphoproliferative disease that is characterized by widespread vascular lesions. Most studies that evaluated the viral load in tissues of animals with SA-MCF were done in the Northern Hemisphere, with scant information from the Southern part of the globe. This study investigated the viral load of OvGHV2 in the tissues of cattle and an underdeveloped fetus with SA-MCF from three distinct biomes of Brazil. All animals had clinical and histopathological manifestations consistent with SA-MCF. Molecular testing identified the OvGHV2 tegument protein and glycoprotein B genes in the tissues of all animals and the fetus. Viral quantification based on the DNA polymerase gene detected elevated loads of OvGHV2 in tissues with histopathological evidence of SA-MCF and organs with unknown histological data, except for the tissues of the fetus, where the viral load was comparatively reduced. The viral loads detected in multiple organs of cattle from this study with SA-MCF are consistent with those identified in different animal species from the USA and Europe. The detection of a low viral load of OvGHV2 in fetal tissue confirmed transplacental dissemination since elevated viral loads were detected in multiple tissues of the cow with SA-MCF. Furthermore, the elevated viral loads detected in the pulmonary tissues of cattle with interstitial pneumonia indicate that OvGHV2 is an inductor of pulmonary disease in cattle.

Bovine astrovirus and its role in lymphocytic encephalitis in cattle in Ontario, Canada, 1988-2019.

Astroviruses have been found in cattle and other species with encephalitis. Our objective was to determine the frequency of neurotropic bovine astrovirus (BoAstV) in cases of encephalitis in cattle ≥ 4-mo-old. Of 56 cases of idiopathic lymphocytic encephalitis examined retrospectively (1988-2019), fixed brain from 11 cases (19%) tested positive by semi-quantitative RT-PCR for BoAstV CH13/NeuroS1. None of the control cases tested positive, including 32 with other forms of encephalitis and 40 with no neurologic disease. Most astrovirus-positive cases were 1-2-y-old, with a range of 7 mo to 7 y, and affected both beef and dairy breeds with wide geographic distribution. BoAstV-positive cases had acute onset of neurologic signs of 12 h to 7 d before death or euthanasia. Affected cattle had lymphocytic inflammation throughout the brain including cerebrum, thalamus, midbrain, cerebellum, medulla oblongata, and spinal cord, and affecting gray and white matter. Further PCR testing identified a possible cause in 9 of the 45 (20%) remaining idiopathic cases of lymphocytic encephalitis, including eastern equine encephalitis virus, Listeria monocytogenes, bovine viral diarrhea virus, bovine alphaherpesvirus 1, and ovine gammaherpesvirus 2 (malignant catarrhal fever); we found no cases of infection by West Nile virus, rabies virus, or Chlamydia spp. No cause was identified in 36 of 56 (64%) cases of lymphocytic encephalitis. We frequently identified neurotropic BoAstV in cases of lymphocytic encephalitis that had no previously identified cause. Neurotropic BoAstV infections had gone undetected for decades, but the frequency of BoAstV infections has not increased among contemporary cases.

Viral Diseases in Water Buffalo (Bubalus bubalis): New Insights and Perspectives.

The water buffalo (Bubalus bubalis) has great adaptability to rustic environments and more variable conditions than cattle, who generally share the habitat. Diseases carried by buffaloes are relatively unknown and ignored and could be transmissible; an imbalance occurs between pathogens, environment, and susceptible hosts, generating a severe animal health problem. Also relevant is the effect of climate change on the populations of vectors that transmit viral diseases. The discovery of new virus variants that can pass from bovine (Bos) to buffalo or vice versa or to humans has highlighted the relevance of viruses crossing the host barrier. This review discusses the clinical viral diseases most reported in the water buffalo, characteristics, epidemiology, and recent findings about disease behavior, interaction with other species, the host, vectors, and pathogens. Diseases reviewed include Foot and Mouth Disease, Rinderpest, Malignant Catarrhal Fever, Infectious Bovine Rhinotracheitis, Bovine Viral Diarrhea, and Rabies. Also, vector-borne diseases include Lumpy Skin Disease, Ephemeral Fever, and Blue Tongue. The review also considers emerging viruses such as Buffalo Pox and Schmallenberg and, finally, other viruses such as papillomatosis. The knowledge and epidemiology of buffalo viral diseases must be constantly reconsidered and updated for adequate prevention and control programs.

Sheep-associated malignant catarrhal fever in cattle: an update

Sheep-associated malignant catarrhal fever (SA-MCF) is a frequently fatal disease of many species including cattle, caused by ovine herpes virus-2. Sheep are the main reservoir host species and transmit the virus to other species, usually via aerosol. Clinical signs in affected animals can impact various body systems, and infection can be sub-clinical and become latent. No specific treatments exist, and empirical treatments are generally unrewarding. Occasionally some animals may survive, but there remains a chronic arteriopathy. Currently, the disease is controlled on farms is by reducing contact between susceptible hosts and sheep as much as possible, although a vaccine would be a useful protective strategy should one be developed.

Ovine Herpesvirus 2 Glycoprotein B Complementation Restores Infectivity to a Bovine Herpesvirus 4 gB-Null Mutant.

Ovine herpesvirus 2 (OvHV-2) and bovine herpesvirus 4 (BoHV-4) are gamma herpesviruses that belong to the genera Macavirus and Rhadinovirus, respectively. As with all herpesviruses, both OvHV-2 and BoHV-4 express glycoprotein B (gB), which plays an essential role in the infection of host cells. In that context, it has been demonstrated that a BoHV-4 gB-null mutant is unable to infect host cells. In this study, we used homologous recombination to insert OvHV-2 ORF 8, encoding gB, into the BoHV-4 gB-null mutant genome, creating a chimeric BoHV-4 virus carrying and expressing OvHV-2 gB (BoHV-4∆gB/OvHV-2-gB) that was infectious and able to replicate in vitro. We then evaluated BoHV-4∆gB/OvHV-2-gB as a potential vaccine candidate for sheep-associated malignant catarrhal fever (SA-MCF), a fatal disease of ungulates caused by OvHV-2. Using rabbits as a laboratory model for MCF, we assessed the safety, immunogenicity, and efficacy of BoHV-4∆gB/OvHV-2-gB in an immunization/challenge trial. The results showed that while BoHV-4∆gB/OvHV-2-gB was safe and induced OvHV-2 gB-specific humoral immune responses, immunization conferred only 28.5% protection upon challenge with OvHV-2. Therefore, future studies should focus on alternative strategies to express OvHV-2 proteins to develop an effective vaccine against SA-MCF.

Malignant catarrhal fever: pathogenesis, problems of prevention and control (review)

Malignant catarrhal fever is a non-contagious infectious disease that occurs sporadically in the population of cattle and wild artiodactyls. The source of the causative agent of the disease is wildebeest, sheep, and goats, in which the infectious process is usually asymptomatic. The manifestation of clinical and pathological signs is variable, often characterized by damage to the nervous, respiratory and digestive systems, in the organs and tissues of which signs of lymphocytic and histiocytic vasculitis are noted. Diagnosis of the disease is carried out comprehensively using clinical and histological methods, but the main method is the detection of viral DNA in the material using PCR. However, control and prevention of the disease have a number of difficulties due to insufficient efficiency of diagnostic measures, lack of vaccines and outdated regulatory framework. There are no commercially available kits for diagnosing the disease in Russia. This diagnosis is available to research organizations, where complex research protocols are used. Due to insufficient knowledge of the pathogenesis, attempts to create effective vaccines against malignant catarrhal fever have not yet been successful, which does not allow for the full implementation of preventive measures. Regulatory acts defining approaches to control and elimination of the disease have not been updated for more than 20 years. The complex of these problems significantly distorts the objective epizootic situation on malignant catarrhal fever and contributes to its hidden spread with unclear economic and economic damage.

Identification and characterisation of Gamma-herpesviruses in zoo artiodactyla

BackgroundViruses within the γ-herpesviruses subfamily include the causative agents of Malignant Catarrhal Fever (MCF) in several species of the order Artiodactyla. MCF is a usually fatal lymphoproliferative disease affecting non-adapted host species. In adapted host species these viruses become latent and recrudesce and transmit during times of stress or immunosuppression. The undetected presence of MCF-causing viruses (MCFVs) is a risk to non-adapted hosts, especially within non-sympatric zoological collections. This study investigated the presence of MCFVs in six different zoological collections in the UK, to evaluate the presence of subclinical/latent MCFVs in carrier animals.MethodsOne-hundred and thirty eight samples belonging to 54 different species of Artiodactyla were tested by Consensus Pan-herpes PCR. The positive samples were sequenced and subjected to phylogenetic analyses to understand their own evolutionary relationships and those with their hosts.ResultsTwenty-five samples from 18 different species tested positive. All viruses but one clustered in the γ-herpesvirus family and within the Macavirus as well as the non-Macavirus groups (caprinae and alcelaphinae/hippotraginae clusters, respectively). A strong association between virus and host species was evident in the Macavirus group and clustering within the caprinae group indicated potential pathogenicity.ConclusionThis study shows the presence of pathogenic and non-pathogenic MCFVs, as well as other γ-herpesviruses, in Artiodactyla species of conservation importance and allowed the identification of new herpesviruses in some non-adapted species.

Coding complete genome of a strain of ovine herpesvirus-2 associated with a clinical case of malignant catarrhal fever in a domestic lamb.

Ovine herpesvirus-2 causes sheep-associated malignant catarrhal fever, a fatal disease of ruminants and pigs. The virus is carried by sheep, and infection is typically subclinical. Here, we report the coding complete genome sequence of a strain of OvHV-2 obtained from a clinically affected domestic lamb.

Molecular and phylogenetic analysis of herpesviruses in endangered free-ranging cervids of Chile: ovine gammaherpesvirus-2-A novel threat to wild and domestic animal health in Chilean Patagonia.

Herpesvirus infections have been highlighted as emerging diseases affecting wildlife health and the conservation of several taxa. Malignant catarrhal fever (MCF) and infectious keratoconjunctivitis (IKC) are two viruses that infect wild ruminants. Nevertheless, epidemiological data on herpesviruses in South American wild ruminants are limited. An outbreak of caprine gammaherpesvirus-2 (CpHV-2) that recently was suspected as the cause of MCF in southern pudus (Pudu puda) prompted the need to conduct molecular screenings in Chilean cervids to understand the epidemiology of herpesviruses. The aim of this study was to determine the occurrence and genetic diversity of herpesviruses in free-ranging cervids from Chile. Herpesvirus infection was assessed in antemortem blood samples (n = 86) from pudus (n = 81) and huemuls (Hippocamelus bisulcus) (n = 5), as well as postmortem samples of spleen (n = 24) and lung (n = 3) from pudus, using a nested pan-herpesvirus PCR assay. Combining all suitable sample types, DNA of pudu gammaherpesvirus-1 was detected in five pudues and five huemuls, with an overall prevalence of 9.90% (n = 10/101; 95% CI = 5.11-17.87%). One pudu tested positive for ovine gammaherpesvirus-2 (n = 1/96; 1.04%; 95% CI = 0.05-6.49%), and one pudu tested positive for a Macavirus sequence with 98.63 similarity to ovine gammaherpesvirus-2 (n = 96; 1.04%; 95% CI = 0.05-6.49%). To the best of our knowledge, this is the first report of a herpesvirus in huemul and of ovine gammaherpesvirus-2 in Chile. Our results also confirm the active circulation of herpesvirus in free-ranging cervids in Chilean Patagonia, and as such, MCF should be considered as a possible cause of disease in free-ranging Chilean pudus and livestock species. Further research is necessary to develop a plan of systematic monitoring (serological and pathological screening) of herpesviruses in Chilean wild and domestic ruminants to understand their diversity and impact on animal health and conservation.

Malignant catarrhal fever in a goat: manifestation of virus-induced erythema multiforme.

Malignant catarrhal fever (MCF), caused by ovine herpesvirus 2 (OvHV2; Orthoherpesviridae, Macavirus ovinegamma2), has sheep as natural hosts. OvHV2 is an important macavirus globally that induces fatal disease in dead-end hosts. Goats, which can be infected subclinically with OvHV2, rarely develop MCF. A 28-wk-old female goat was presented with fever and multifocal crusty skin lesions. Histologic examination of a skin biopsy suggested erythema multiforme (EM), with pyoderma and dermal vasculitis. The doe was euthanized and subjected to postmortem and histologic examination. MCF was suspected and PCR assays for macaviruses were performed, followed by immunohistochemistry (IHC) for OvHV2 latency-associated nuclear antigen (oLANA), RNA in situ hybridization for Ov2.5 mRNA, and IHC to characterize infiltrating leukocytes. The main postmortem finding was severe multifocal ulcerative dermatitis with macrophage- and T cell-mediated arteritis. The latter was also detected in kidney, spleen, heart, and intestinal wall. The PCR assay detected high loads of OvHV2 in tissues. OvHV2 oLANA and Ov2.5 mRNA were expressed within the lesions in leukocytes, endothelial cells, fibroblasts, and/or keratinocytes. Our case confirms that MCF can initially manifest clinically as a skin disease in goats and as EM with confirmed viral etiology.

Molecular identification of bovine gammaherpesvirus 6 in water buffaloes (Bubalus bubalis) with histological evidence of pulmonary and intestinal lesions

Bovine gammaherpesvirus 6 (BoGHV6), formerly known as bovine lymphotropic virus, belongs to the genus Macavirus, which includes the porcine lymphotropic herpesviruses (PLHV) and members collectively referred to as malignant catarrhal fever virus (MVFV) group. This study investigated the possible participation of BoGHV6 in the development of intestinal and pulmonary diseases of buffaloes. Intestinal and pulmonary fragments were obtained from 19 buffaloes from Goiás, Central-western Brazil. Histopathology revealed interstitial pneumonia in all pulmonary fragments evaluated, while two of these had suppurative bronchopneumonia. Furthermore, all intestinal fragments demonstrated atrophic enteritis. BoGHV6 DNA was amplified from 31.6% (6/19) of the buffaloes with interstitial pneumonia and in 26.3% (5/19) with atrophic enteritis. The phylogenetic evaluation revealed that the strain of BoGHV6 amplified from these buffaloes had 100% nucleotide (nt) sequence identity with the reference strains of BoGHV6, but only 69.5 to 73.7% and 65.8 – 69.5% nt sequence identity with members of the MCFV and the PLHV, respectively. Additionally, molecular assays to detect important pathogens of ruminants did not amplify the respective nucleic acids in the tissues evaluated. These results demonstrated that BoGHV6 was amplified from tissues of buffaloes with histopathologic diagnoses of interstitial pneumonia and atrophic enteritis, suggesting the possible participation of this virus in the development of the intestinal and pulmonary lesions herein identified.

Ivermectin inhibits replication of the malignant catarrhal fever virus alcelaphine herpesvirus 1

Malignant catarrhal fever is a lymphoproliferative disease of cattle and other ungulates that is caused by genetically and antigenically related gamma herpesviruses of the genus Macavirus. Infection of the natural host species is efficient and asymptomatic but spread to susceptible hosts is often fatal with clinical signs including fever, depression, nasal and ocular discharge. There is no recognised treatment for MCF but a vaccine for one MCF virus, alcelaphine herpesvirus 1 (AlHV-1), has been described. In this paper we describe the inhibition of AlHV-1 replication and propagation by the anthelminthic drug ivermectin. Concentrations of 10 μM or greater led to significant reductions in both copy number and viable titre of virus tested in culture medium, with little replication detected at over 20 μM ivermectin. In the absence of alternative treatments, further testing of ivermectin as a candidate antiviral treatment for MCF may therefore be justified.

Case report: Systemic granulomatous disease with vasculitis in a bull due to hairy vetch (Vicia villosa) toxicosis

A 5-year-old Angus bull presented for generalized alopecia, thickened pleated skin and profuse malodorous watery diarrhea. This bull, along with 6 other beef bulls, had been grazing in a pasture containing hairy vetch (Vicia villosa) for 5-6 months. Diagnostic samples included blood for CBC and chemistry, skin biopsy and scrapings, fecal sample and liver biopsy. Biopsy of skin revealed a chronic, moderate, perivascular and perifollicular, eosinophilic to histiocytic-lymphocytic dermatitis with moderate acanthosis and hyperkeratosis indicative of an ongoing hypersensitivity reaction. The bull did not respond to treatment and was euthanized due to poor prognosis. At necropsy, disseminated systemic granulomatous disease with variable numbers of multinucleated giant cells and eosinophils was observed in multiple organs indicative of systemic granulomatous disease, along with vasculitis in the kidney, liver, and adrenal gland, indicative that vasculitis is involved in the pathogenesis of the lesions. Viral diseases were ruled out by immunohistochemistry of sections of kidney and by in situ hybridization on kidney sections. CD3, CD20, and CD79a lymphocyte markers immunostaining of sections of kidney revealed predominance of CD3 positive lymphocytes, indicative of predominance of T-cell lymphocytes supporting a type-IV hypersensitivity. The vasculitis observed prompted formulating a list of differential diagnoses for infectious diseases that have evidence of vasculitis, including bovine virus diarrhea virus and malignant catarrhal fever. Vasculitis, although previously described in cases of hairy vetch poisoning, is not observed in every single case of the disease. The remaining bulls were not removed from the pasture and none developed clinical signs.