Immunohistochemical identification of a malignant catarrhal fever virus in cattle with renal diseases from Paraná state, Southern Brazil: a retrospective epidemiological study.

Malignant catarrhal fever (MCF) is a viral infectious disease caused by specific members of the Macavirus genus that are referred to as the MCF virus (MCFV) complex group. This study determined the prevalence of MCFV-associated infections in cattle within the mesoregions of the state of Paraná, Southern Brazil, by analyzing the histopathologic patterns of renal lesions in association with positive immunoreactivity to intralesional antigens of MCFV. Intracytoplasmic MCFV antigens were identified in 41.7% (48/115) of the kidneys of cattle evaluated. Lymphocytic interstitial nephritis, vascular degeneration, and ballooning degeneration of the renal tubules were the principal histopathological findings associated with positive immunoreactivity to MCFV. The results indicate that MCFV infections are endemic within the state of Paraná and suggest that the kidney can be of diagnostic value in suspected cases of MCF-associated infections in cattle. Furthermore, the utilization of an in situ diagnostic technique resulted in the detection of a greater number of cases of infections by MCFV than previously identified using other diagnostic methods. Additionally, degenerative vascular lesions of the kidney should be considered during the establishment of a histological diagnosis of MCFV-induced infections in cattle in the absence of fibrinoid change or necrotizing vasculitis.

First identification and characterization of ovine gammaherpesvirus type 2 in horses and artiodactyla from an outbreak of malignant catarrhal fever in Mexico.

Ovine gammaherpesvirus 2 (OvHV-2), a member of the genus Macavirus, causes sheep-associated malignant catarrhal fever (SA-MCF), a fatal lymphoproliferative disease affecting a wide variety of ungulates in addition to horses. This study described an outbreak of SA-MCF in Mexico and the identification of the OvHV-2 virus in primary rabbit testis cultures through the generation of intranuclear inclusion bodies, syncytia, immunofluorescence (IF), immunocytochemistry (ICC), immunohistochemistry (IHC), endpoint polymerase chain reaction (PCR), and partial sequencing of the ORF75 gene. The animals involved in this outbreak showed mucogingival ulcers in the vestibule of the mouth and tongue, hypersalivation, corneal opacity, reduced food consumption, and weight loss of variable severity. These clinical signs and the histopathological findings suggested the diagnosis of SA-MCF. Buffy coat fractions from the anticoagulated blood samples of ill animals were collected and analyzed by PCR. Positive buffy coats were used to inoculate the primary cell cultures of rabbit testis to identify the virus. Small clusters of refractile cytomegalic cells, characteristic of viral cytopathic effects, were observed between 48 and 72 h post-infection. Furthermore, intranuclear acidophilic inclusion bodies (IBs) were identified in the inoculated primary culture cells, and the cytoplasm showed immunoreactivity with hyperimmune rabbit serum against OvHV-2. Moreover, in the liver histological sections from sick deer, immunoreactive juxtanuclear IBs were identified with the same rabbit hyperimmune serum. The obtained sequences were aligned with the OvHV-2 sequences reported in GenBank and revealed a nucleotide identity higher than 98%. Based on the evidence provided in this study, we conclude that the outbreak of SA-MCF in the municipality of Tequisquiapan in the state of Queretaro, Mexico, was caused by OvHV-2. This is the second study reporting that horses are susceptible to OvHV-2 infection and can develop SA-MCF. We identified for the first time in Mexico, the presence of OvHV-2 in buffy coats from horses and Artiodactyla.

A review of the epidemiological, clinical, and pathological aspects of malignant catarrhal fever in Brazil.

Sheep-associated malignant catarrhal fever (SA-MCF), the form of MCF that occurs in Brazil, is a severe, frequently fatal, infectious disease caused by ovine gammaherpesvirus-2 (OvHV-2), in which sheep are the asymptomatic hosts and cattle and other cloven-hoofed animals are the accidental hosts. This review provides a critical analysis of the historical, epidemiological aspects and the estimated economic impacts associated with SA-MCF in Brazil. Moreover, the clinical manifestations and pathological lesions associated with SA-MCF in cattle are reviewed and discussed and the phylogenetic distribution of OvHV-2 in Brazil is presented. OvHV-2 is the only MCF virus identified in animals from Brazil. It is recommended that a histopathologic diagnosis of SA-MCF be based on all aspects of vascular disease in the affected animal and not only lymphocytic/necrotizing vasculitis and/or fibrinoid change. Conformation of the intralesional participation of OvHV-2 in these alterations can be achieved by immunohistochemistry and/or in situ hybridization assays. Additionally, it is proposed that OvHV-2 should be considered as a possible infectious disease agent associated with the development of bovine respiratory disease in cattle. Furthermore, the possible role of the small intestine in the dissemination of OvHV-2 is discussed.

Malignant catarrhal fever: An emerging yet neglected disease in captive sika deer (Cervus nippon) herds in China.

Malignant catarrhal fever (MCF) is a fatal lymphoproliferative disease that represents a serious problem in the deer-rearing industry. To better understand an MCF-like disease that has emerged in northern China since 2015, we investigated ten cases by documenting clinical and epidemiological data and analysing causative agents and histopathological changes. In addition, a retrospective screen for Macavirus DNA and a questionnaire-based survey were conducted. Epizootic MCF in Chinese sika deer herds has emerged with a low morbidity of 3.8% (95% CI: 2.5%-5.1%) and a high mortality of 93.2% (95% CI: 86.6%-99.9%). The disease course varied from 3 to 12 days. Aetiologically, OvHV-2 was predominant in the MCFV, accounting for most MCF cases (21/23). In contrast, only two CpHV-2 isolates were phylogenetically closely related to CpHV-2. Diarrhoea and nasal discharges were the most frequent manifestations, although clinical signs varied in some cases. Pathologically typical lesions of haemorrhage, necrosis and lymphoid cell infiltration were readily observed in a variety of organs. Vasculitis caused by vascular and perivascular lymphoid cell infiltration was common. The retrospective survey suggested a low positive rate (3/275) of MCFV DNA in peripheral blood lymphocytes (PBL). The questionnaire-based survey suggested the disease was neglected by local veterinarians, who did not acknowledge the risk of co-rearing deer with reservoir species. Collectively, the emerging epizootic MCF in Chinese sika deer herds remains neglected, emphasizing the urgency of initiating full-field diagnoses and control strategies.

Quantification of ovine herpesvirus 2 by digital PCR in an outbreak of malignant catarrhal fever.

Infection with ovine gammaherpesvirus 2 (OvHV-2) is generally asymptomatic in sheep; however, when it crosses the species barrier, it causes malignant catarrhal fever (MCF) in cattle. In the present study, we developed a real-time PCR assay and a droplet digital PCR assay and use both methods to study an outbreak caused by OvHV-2. Both PCR methods showed high sensitivity and specificity and were able to detect low copy numbers of OvHV-2 in sheep and cattle. The present study describes the first digital PCR quantification of OvHV-2 genome copies in samples collected from sheep and cattle.

Observations of mortality in farmed bison in the Canadian prairies: 2103 - 2016.

The present study is a continuation of a previous mortality study on Saskatchewan bison farms with special emphasis on Malignant Catarrhal Fever. The updated objective of the study was to estimate the most common causes of mortality in farmed bison herds in Western Canada. Results were compared to the previous Saskatchewan study to assess the similarities and differences in the etiology associated with farmed bison deaths across the Prairie Provinces of Canada. The most common cause of death was respiratory disease associated with Mycoplasma bovis, although this was restricted to Alberta and Saskatchewan farm locations. This was in contrast to the previous Saskatchewan based study which did not identify any deaths involving this pathogen. An updated overall assessment of the risks of Malignant Catarrhal Fever in farmed bison at various proximities to sheep operations further confirmed the low risk of occurrence on farms within a 1 km boundary fence distance.

Seroprevalence of malignant catarrhal fever virus in captive wildebeest (Connochaetes sp.) in France.

Alcelaphine herpesvirus 1 (AlHV-1) is a gammaherpesvirus carried asymptomatically by wildebeests (Connochaetes sp.) in sub-Saharan Africa. Although asymptomatic in wildebeest, AlHV-1 infection in a number of other ruminant species causes a severe and fatal lymphoproliferative disease named wildebeest-derived malignant catarrhal fever (WD-MCF). Several endangered species of captive ruminants are highly susceptible to developing WD-MCF if infected by AlHV-1, which is a critical concern in zoos, game reserves and wildlife parks where wildebeests are also kept in captivity. Here, we investigated the seroprevalence of AlHV-1 in 52 captive wildebeests randomly sampled from five different zoos in France. We found 46% (24/52) seropositive animals and detected AlHV-1 DNA in one of them, demonstrating that AlHV-1 infection is present in captive wildebeests in France. In an interesting manner, the repartition of seropositive wildebeests was not homogenous between zoos with 100% (20/20) of seronegative animals in three parks. These results further highlight the importance of considering WD-MCF as a threat for clinically susceptible species and encourage for testing AlHV-1 infection in captive wildebeests as a management control strategy.

Caprine herpesvirus 2-associated malignant catarrhal fever of captive sika deer (Cervus nippon) in an intensive management system.

BACKGROUND: Caprine herpesvirus 2 (CpHV-2) infection usually induces chronic malignant catarrhal fever (MCF) in sika deer (Cervus nippon), with the primary signs of weight loss, dermatitis and alopecia. CASE PRESENTATION: Here, we report a case of CpHV-2-associated acute MCF in a sika deer herd raised in an intensive management system distant to the reservoir goats. Affected deer developed clinical signs of high fever (41 °C) followed by nasal discharge and lameness. Severe lesions of hemorrhage, necrosis and infiltration of lymphoid cells could readily be observed in the lung, kidney, heart valves and subcutaneous tissue surrounding a tendon. Etiologically, identical CpHV-2 specific DNA sequences were detected in peripheral blood lymphocyte (PBL) from the affected deer and reservoir goats. CONCLUSION: In summary, domestic goats were the reservoir of the CpHV-2, which is the causative agent of the outbreak of MCF in the three hinds. The disease was probably transmitted via aerosol infection. In addition, necrosis and inflammation in subcutaneous tissue surrounding a tendon was the reason for lameness. Therefore, MCF should be put into a differential diagnostic list when similar disease occurs in sika deer herds.

Taxa de infecção pelo Herpesvírus ovino tipo 2 (OvHV-2) em rebanhos de ovinos no Distrito Federal / Ovine herpesvirus type 2 (OvHV-2) infection rate in sheep herds of the Federal District, Brazil

A febre catarral maligna (FCM) é uma doença causada pela infecção de bovinos pelo herpesvírus ovino tipo 2 (OvHV-2), responsável por perdas econômicas em diferentes regiões do Brasil. Neste trabalho descreve-se a detecção molecular por nested-PCR (nPCR) do OvHV-2 em amostras de secreção/esfoliação nasal e fração celular sanguínea (FCS) de ovinos provenientes de 8 propriedades do Distrito Federal. Das 188 amostras nasais analisadas, 88 (41,5%) foram positivas. Ovelhas prenhes não apresentaram diferenças na taxa de infecção em comparação com fêmeas paridas. Fêmeas recém-paridas apresentaram taxa de infecção pelo OvHV-2 maior que em animais que pariram há mais de 60 dias. Amostras de secreção/esfoliação nasal permitiram a detecção por nPCR de animais infectados com uma eficiência aproximadamente duas vezes maior que em amostras de fração celular sanguínea. No Brasil, informações epidemiológicas sobre a infecção pelo OvHV-2 nos rebanhos ovinos e fatores envolvidos no surgimento de surtos de FCM em bovinos são escassos. Este estudo pode servir de subsídio para elucidar as características da enfermidade e para novos estudos sobre a epidemiologia da doença no Distrito Federal e em outros Estados do Brasil.(AU)

Malignant catarrhal fever (MCF) is a disease caused by bovine infection with ovine herpesvirus type 2 (OvHV-2) and responsible for economic losses in different Brazilian regions. This paper describes the molecular detection of OvHV-2 by nested-PCR (nPCR) in nasal secretion/exfoliation samples and blood cell fraction (BCF) of sheep from 8 properties in the Federal District. Among the 188 nasal samples, 88 (41.5%) were positive to OvHV-2. Pregnant ewe presented no differences at the infection rate in comparison with parous females. Newly calved sheep showed higher OvHV-2 infection rate than female over 60 days of calving. Nasal samples allowed the detection of infected animals by nPCR with efficiency about twice than that in the blood cell fraction samples. In Brazil, epidemiological information about OvHV-2 infection in sheep flocks and factors involved in emergence of FCM outbreaks in cattle are still scarce. This study may provide support for elucidating some characteristics of the disease and for further epidemiological studies in the Federal District and other Brazilian States.(AU)

Wildlife: topical issues, recent cases.

This focus article was prepared by Paul Duff, Paul Holmes and Alex Barlow of the APHA Wildlife Expert Group.

Genetic characterization of ovine herpesvirus 2 strains involved in water buffaloes malignant catarrhal fever outbreaks in Southern Italy.

Ovine herpesvirus 2 (OvHV-2) was responsible for two outbreaks of malignant catarrhal fever (MCF) on two water buffalo farms in Southern Italy. In this study, the presence of this virus in the nasal swabs from sick animals as well as in the organs of dead buffaloes was ascertained by a Real-time PCR assay. Positive samples also underwent a relative quantitative analysis of the viral DNA in them. All the dead animals had the highest relative viral quantities, while buffaloes recovering from the virus had intermediate quantities, and asymptomatic OvHV-2-positive sheep had the lowest relative quantities (as compared with the calibrator). The strains involved in the MCF outbreaks underwent genetic characterization by sequencing segments of their ORF50, ORF75 and Ov9.5 genes. The results showed that the outbreaks were caused by two specific genetic variants of OvHV-2, and that these variants exhibit nucleotide differences at the loci analysed. Sheep living in the surrounding farms, as well as sheep kept with buffaloes, were also investigated as possible transmitters of the virus. In this regard, local strategies for the control of MCF should consider separating reservoir species from susceptible animals.

An observational study of mortality on bison farms in Saskatchewan with special emphasis on malignant catarrhal fever.

In December 2011, the Malignant Catarrhal Fever (MCF) Task Force in Saskatchewan recommended that research be conducted on the relationship between the proximity of bison and sheep under typical commercial production settings and bison deaths due to MCF. The objective of this study was to evaluate all causes of death in bison herds and compare the incidence of MCF in herds at varying distances of exposure from sheep operations. Necropsies were completed on 76 of 133 bison reported to have died during the 18-month study period. A total of 7 MCF deaths was reported from 2 large herds within 1.0 km of sheep operations. Although there was a greater risk of MCF deaths in bison herds within 1.0 km of sheep operations than in herds more than 1.0 km away, the overall incidence of MCF deaths within the study period was very low. Most deaths were attributed to non-infectious causes, including copper deficiency.

Étude observationnelle de la mortalité dans des fermes de bisons en Saskatchewan, avec une emphase particulière sur la fièvre catarrhale maligne des bovins. En décembre 2011, le groupe de travail de la Saskatchewan sur la fièvre catarrhale maligne des bovins (FCM) a recommandé la réalisation de travaux de recherche pour étudier le lien entre la proximité des bisons et des moutons dans des milieux de production commerciaux typiques et la mortalité des bisons attribuable à la FCM. La recherche avait pour but d'évaluer toutes les causes de mortalité dans les troupeaux de bisons, puis de comparer l'incidence de la FCM dans les troupeaux à diverses distances d'exposition des exploitations d'élevage de moutons. Des nécropsies ont été réalisées sur 76 des 133 bisons dont la mort a été signalée durant la période d'étude de 18 mois. Un total de sept morts causées par la FCM a été signalé dans deux grands troupeaux situés à une distance de moins de 1 km. Le risque de mortalité pour cause de FCM était supérieur dans les troupeaux de bisons situés à moins de 1 km des exploitations de moutons que dans les troupeaux situés à une distance de plus de 1 km. Cependant, l'incidence totale de mortalité causée par la FCM était très faible. La plupart des mortalités étaient attribuables à des causes non infectieuses, y compris une carence en cuivre.(Traduit par Isabelle Vallières).

An Acute Multispecies Episode of Sheep-Associated Malignant Catarrhal Fever in Captive Wild Animals in an Italian Zoo.

In July 2011, in a zoological garden in Rome, Italy, malignant catarrhal fever (MCF), a fatal, systemic disease of Artiodactyla, was suspected on the basis of neurological signs and gross lesions observed in a banteng, the first animal to die of this infection. An MCF type-specific PCR with subsequent sequencing of the PCR amplicon confirmed the aetiological agent as ovine herpesvirus-2 (OvHV-2). Biological samples were collected from the dead animals for gross, histological, bacteriological, virological and serological examinations. An epidemiological investigation was conducted to identify the source of the outbreak, as further deaths due to OvHV-2 still occurred after the removal of the acknowledged reservoirs, domestic sheep and goats. For this purpose, samples from other susceptible species and reservoir hosts were collected for virological and serological analysis. In conjunction, a retrospective sero-investigation was conducted on sera collected between 1999 and 2010 from some of the species involved in the present episode. In total, 11 animals belonging to four different species (banteng, Himalayan tahr, Nile lechwe and sika deer) died between July 2011 and October 2012. The severe gross and histological lesions were consistent with the disease, namely haemorrhages and congestion of several organs as well as lymphoid cell infiltrates and vasculitis of varying severity. The virological tests confirmed that all animals had died of sheep-associated MCF. The investigation indicated that the OvHV-2 infection could have been due to the arrival of sheep in the petting zoo, with cases commencing after first lambing and subsequent shedding of virus. This was also supported by the serological retrospective study that indicated limited previous MCF virus circulation. Further MCF cases that occurred even after the removal of the domestic sheep and goats were attributed to the mouflon. This episode confirms the importance of biosecurity measures in zoos, which house MCF susceptible species, especially those endangered.

Alcelaphine Herpesvirus-1 (Malignant Catarrhal Fever Virus) in Wildebeest Placenta: Genetic Variation of ORF50 and A9.5 Alleles.

Alcelaphine herpesvirus-1 (AlHV-1), a causative agent of malignant catarrhal fever in cattle, was detected in wildebeest (Connochaetes taurinus) placenta tissue for the first time. Although viral load was low, the finding of viral DNA in over 50% of 94 samples tested lends support to the possibility that placental tissue could play a role in disease transmission and that wildebeest calves are infected in utero. Two viral loci were sequenced to examine variation among virus samples obtained from wildebeest and cattle: the ORF50 gene, encoding the lytic cycle transactivator protein, and the A9.5 gene, encoding a novel polymorphic viral glycoprotein. ORF50 was well conserved with six newly discovered alleles differing at only one or two base positions. In contrast, while only three new A9.5 alleles were discovered, these differed by up to 13% at the nucleotide level and up to 20% at the amino acid level. Structural homology searching performed with the additional A9.5 sequences determined in this study adds power to recent analysis identifying the four-helix bundle cytokine interleukin-4 (IL4) as the major homologue. The majority of MCF virus samples obtained from Tanzanian cattle and wildebeest encoded A9.5 polypeptides identical to the previously characterized A9.5 allele present in the laboratory maintained AlHV-1 C500 strain. This supports the view that AlHV-1 C500 is suitable for the development of a vaccine for wildebeest-associated MCF.

The epidemiology of malignant catarrhal fever (MCF) and contribution to cattle losses in farms around Rhodes Matopos National Park, Zimbabwe.

Malignant catarrhal fever (MCF) is a fatal viral disease of cattle prevalent in farms bordering wildlife conservancies and national parks where cattle and wildebeests cohabit. The behaviour of the diseases and its related contribution to cattle mortalities has been poorly studied in the smallholder sector. A study was therefore conducted to established trends of occurrence and contribution to animal losses in 97 smallholder farms and eight commercial farms bordering Rhodes Matopos National Park (RMNP). A structured questionnaire and secondary records were used to gather the data in the smallholder and commercial farms, respectively. In the smallholder sector, cattle farmers from three villages were chosen as respondents in a survey to provide information on the impact and extent of losses in their cattle herds. Epidemiological data from January 2006 to April 2014 was extracted from farm records from two Matopos Research Institute farms. A seasonal pattern in MCF occurrence was observed. Most cattle mortalities were recorded during summer (February to May), with highest mortalities recorded from March to April. Sporadic cases were however recorded in October and November in year 2007, 2010, and 2012. The study further revealed that MCF caused 71 and 21 % of cattle mortalities in the commercial and smallholder sector, respectively. Deferred grazing of wildebeest hotspots by restriction of cattle movement during summer or November to April period avoids wildebeest-cattle contact. This can alleviate cattle mortalities due to MCF.

A geographic cluster of malignant catarrhal fever in moose (Alces alces) in Norway.

Three cases of lethal sheep-associated malignant catarrhal fever (SA-MCF) in free-ranging moose (Alces alces) were diagnosed in Lesja, Norway, December 2008-February 2010. The diagnosis was based on PCR identification of ovine herpesvirus 2 (OvHV-2) DNA (n = 3) and typical histopathologic lesions (n = 1). To study the possibility of subclinical or latent MCF virus (MCFV) infection in this moose population and in red deer (Cervus elaphus), we examined clinically normal animals sampled during hunting in Lesja 2010 by serology and PCR. Sera from 63 moose and 33 red deer were tested for antibodies against MCFV by competitive-inhibition enzyme-linked immunosorbent assay. To test for MCFVs, a consensus PCR for herpesviral DNA was run on spleen samples from 23 moose and 17 red deer. All samples were antibody and PCR negative. Thus, there is no evidence of previous exposure, subclinical infection, or latent infection in this sample. This seasonal cluster of SA-MCF cases (2008-10) may be attributable to exposure of moose to lambs when OvHV-2 shedding is presumed to be maximal, compounded by an unusual extended grazing period by sheep in the autumn.

The economic impact of malignant catarrhal fever on pastoralist livelihoods.

This study is the first to partially quantify the potential economic benefits that a vaccine, effective at protecting cattle against malignant catarrhal fever (MCF), could accrue to pastoralists living in East Africa. The benefits would result from the removal of household resource and management costs that are traditionally incurred avoiding the disease. MCF, a fatal disease of cattle caused by a virus transmitted from wildebeest calves, has plagued Maasai communities in East Africa for generations. The threat of the disease forces the Maasai to move cattle to less productive grazing areas to avoid wildebeest during calving season when forage quality is critical. To assess the management and resource costs associated with moving, we used household survey data. To estimate the costs associated with changes in livestock body condition that result from being herded away from wildebeest calving grounds, we exploited an ongoing MCF vaccine field trial and we used a hedonic price regression, a statistical model that allows estimation of the marginal contribution of a good's attributes to its market price. We found that 90 percent of households move, on average, 82 percent of all cattle away from home to avoid MCF. In doing so, a herd's productive contributions to the household was reduced, with 64 percent of milk being unavailable for sale or consumption by the family members remaining at the boma (the children, women, and the elderly). In contrast cattle that remained on the wildebeest calving grounds during the calving season (and survived MCF) remained fully productive to the family and gained body condition compared to cattle that moved away. This gain was, however, short-lived. We estimated the market value of these condition gains and losses using hedonic regression. The value of a vaccine for MCF is the removal of the costs incurred in avoiding the disease.

Malignant Catarrhal Fever: An Emerging Disease in the African Buffalo (Syncerus caffer).

Within the tribe Bovini in the subfamily Bovinae, the water buffalo (Bubalus Bubalis), American bison (Bison bison), European bison (Bubalus bonasus) and yak (Bos grunniens) are recognized as species highly susceptible to malignant catarrhal fever (MCF). In contrast, the lack of reports describing clinical MCF in the African buffalo (Syncerus caffer) whether free ranging or captive has led to a perception that African buffaloes are resistant to MCF. During the last decade, several cases of MCF in African buffaloes were confirmed in South Africa and experience with seven of these cases is described in this report. Detection of viral nucleic acid in blood or tissues was successful in six African buffaloes that suffered from clinical signs compatible with MCF. Four were positive for infection with ovine herpesvirus type 2 (the causative virus of sheep-associated MCF), and two were positive for alcelaphine herpesvirus type 1 (causative virus of wildebeest-associated MCF). Histopathological examination of tissue samples from all the animals yielded typical lesions that were consistent with those described for MCF in domestic cattle. Developments in the management of African buffaloes translocated from their traditional habitats have likely contributed to the identification of another susceptible host in the subfamily Bovinae.