The effects of exposure of susceptible alpacas to alpacas persistently infected with bovine viral diarrhea virus.

Reports of bovine viral diarrhea virus (BVDV) infections in alpacas have been increasing in recent years but much is still unknown about the mechanisms of disease in this species. This report characterizes the transmission of BVDV from persistently infected (PI) alpacas to BVDV naïve alpacas, documents shedding patterns, and characterizes the disease effects in both PI and transiently infected alpacas. Two PI alpacas shed BVDV Type 1b virus in most body fluids, and commonly available diagnostic tests verified their status. Bovine viral diarrhea virus Type 1b transient infections produced only mild signs of disease in BVDV naïve alpacas. Viremia was detected in whole blood, but viral shedding during the acute phase was not detected and antibody appeared to be protective upon re-exposure to the virus.

Disseminated Bovine viral diarrhea virus in a persistently infected alpaca (Vicugna pacos) cria.

Bovine viral diarrhea virus (BVDV) is an emerging infectious pathogen of concern to the alpaca industry. A 4-month-old, intact, male alpaca cria was diagnosed as persistently infected with BVDV on the basis of repeated positive antemortem polymerase chain reaction (PCR) and virus isolation (VI) assays and negative serologic titers to BVDV. Immunohistochemistry, real-time reverse transcription PCR, and VI performed on tissues collected at necropsy demonstrated disseminated BVDV-1b infection. Virus was detected in multiple tissues, including parotid salivary gland, testes, prostate, kidneys, skin, and gastrointestinal tract. Demonstration of BVDV in previously unreported tissues suggests additional potential routes of BVDV transmission in alpacas.

Evaluation of a serum ELISA for detection of bovine leukemia viral antibodies in milk samples.

Bovine leukemia virus (BLV) infection has worldwide distribution in both dairy and beef herds. Our study was initiated in order to encourage control of BLV infection by using milk samples, in lieu of serum samples, to readily test lactating animals prior to dry-off and calving. Two Holstein dairy herds (A and B), with known status of BLV infection as determined by serology, were sampled by the collection of serum and fresh milk samples. Selected samples were tested using a USDA-licensed BLV antibody ELISA kit (Bovine leukemia virus antibody test kit; VMRD, Pullman, WA) for serum. Forty-one lactating cows from each herd were sampled. Herd A was confirmed to have endemic BLV infection; herd B was confirmed to be free of BLV infection. The milk ELISA results demonstrated 100% identification of positive and negative animals compared with the serum results. The correlation of the ELISA values between serum and milk samples was 97%, which supports the use of this BLV ELISA on milk samples.

Evidence for persistent Bovine viral diarrhea virus infection in a captive mountain goat (Oreamnos americanus).

Bovine viral diarrhea (BVD) viruses are pestiviruses that have been isolated from domestic and wild ruminants. There is serologic evidence of pestiviral infection in more than 40 species of free-range and captive mammals. Vertical transmission can produce persistently infected animals that are immunotolerant to the infecting strain of Bovine viral diarrhea virus (BVDV) and shed virus throughout their lives. Seven species (white-tailed deer, mouse deer, eland, domestic cattle, alpaca, sheep, and pigs) have been definitively identified as persistently infected with BVDV. This study provides serological, molecular, immunohistochemical, and histological evidence for BVDV infection in 2 captive mountain goats from a zoological park in Idaho. The study was triggered by isolation of BVDV from tissues and immunohistochemical identification of viral antigen within lesions of a 7-month-old male mountain goat (goat 1). Blood was collected from other mountain goats and white-tailed and mule deer on the premises for BVDV serum neutralization, viral isolation, and reverse transcription polymerase chain reaction. One 3-month-old mountain goat (goat 2) was antibody negative and BVDV positive in serum samples collected 3 months apart. This goat subsequently died, and though still antibody negative, BVDV was isolated from tissues and identified by immunohistochemistry within lesions. Sequencing and phylogenetic analysis identified the isolates as BVDV-2. These findings provide evidence of persistent infection in a mountain goat, underscoring the need for pestivirus control strategies for wild ruminants in zoological collections.

Accidental introduction of viruses into companion animals by commercial vaccines.

The use of biologics in veterinary medicine has been of tremendous value in safeguarding our animal populations from debilitating and oftentimes fatal disease. This article reviews the principles of vaccination and the extensive quality control efforts that are incorporated into preparing the vaccines. Examples of adverse events that have occurred in the past and how enhanced vigilance at the level of the veterinarian and the veterinary diagnostic laboratory help to curtail these events are discussed. Emphasis on understanding the ecology of viral infections in dogs and cats is introduced, together with the concepts of the potential role of vaccines in interspecies spread of viruses.

Immortalized sheep microglial cells are permissive to a diverse range of ruminant viruses.

BACKGROUND: Ruminants, including sheep and goats (small ruminants), are key agricultural animals in many parts of the world. Infectious diseases, including many viral diseases, are significant problems to efficient production of ruminants. Unfortunately, reagents tailored to viruses of ruminants, and especially small ruminants, are lacking compared to other animals more typically used for biomedical research. OBJECTIVE: The purpose of this study was to determine the permissibility of a stably immortalized, sheep microglial cell line to viruses that are reported to infect ruminants: bovine viral diarrhea virus (BVDV), bovine herpesvirus 1 (BoHV-1), small ruminant lentiviruses (SRLV), and bovine respiratory syncytial virus (BRSV). METHODS: Sublines A and H of previously isolated, immortalized, and characterized (CD14-positive) ovine microglial cells were used. Bovine turbinate cells and goat synovial membrane cells were used for comparison. Cytopathic changes were used to confirm infection of individual wells, which were then counted and used to calculate the 50% tissue culture infectious dose. Uninoculated cells served as negative controls and confirmed that the cells were not previously infected with these viruses using polymerase chain reaction (PCR). RESULTS: Inoculation of the two microglial cell sublines with laboratory and field isolates of BVDV, BoHV-1, and BRSV resulted in viral infection in a manner similar to bovine turbinate cells. Immortalized microglia cells are also permissive to SRLV, similar to goat synovial membrane cells. CONCLUSION AND CLINICAL RELEVANCE: These immortalized sheep microglial cells provide a new tool for the study of ruminant viruses in ruminant microglial cell line.

A devastating outbreak of malignant catarrhal fever in a bison feedlot.

In early 2003, an outbreak of malignant catarrhal fever (MCF) occurred in a bison feedlot in southern Idaho. The outbreak resulted in a 51.2% (n = 825) mortality rate among bison, which had been exposed to sheep for 19 days. Diagnosis was made by detection of ovine herpesvirus 2 (sheep-associated MCF virus) DNA in tissues or peripheral blood by polymerase chain reaction (PCR), and by histological examination of tissue lesions. Peak losses occurred between 41 and 55 days postmean exposure time (PME), and reached a maximum of 41 head per day. No known cases of MCF were observed among the 177 head of bison that arrived in the lot 3 1/2 weeks after the departure of the sheep. Of the several thousand head of beef cattle in the lot during the outbreak, only a single case of MCF was identified. This outbreak illustrates the devastating impact the MCF virus can have on bison under certain exposure conditions, the high threat posed by adolescent lambs to susceptible species, the significantly greater susceptibility of bison than beef cattle to MCF, and the lack of horizontal transmission from clinically affected bison to herdmates.

Prevalence of vesivirus in a laboratory-based set of serum samples obtained from dairy and beef cattle.

OBJECTIVE: To examine sera obtained from dairy and beef cattle to detect antibodies against vesivirus and compare seroprevalence among cattle within the sample population. SAMPLE POPULATION: Cattle sera from 8 western states and Maryland submitted to the Washington Animal Disease Diagnostic Laboratory during 1999 and 2000. PROCEDURE: Sera were analyzed for vesivirus-specific antibodies by use of a recombinant vesivirus-San Miguel sea lion virus serotype 5-capsid peptide antigen in an indirect ELISA. RESULTS: Overall, 693 sera were tested and 105 (15.2%) had positive results. Seropositive cattle were from 7 states (all cattle from Montana and Maryland 10 and 4, respectively were seronegative). Overall seroprevalence for antivesivirus antibody in herds ranged between 0% and 80% (median, 14%). Higher antibody prevalence was significantly associated with older age, dairy rather than beef cattle, and reasons for submission. Logistic regression of factors (abortion, respiratory tract disease, and all other reasons for sample submission) revealed that older age and other reasons were independently associated with higher seroprevalence. Higher seropositive optical density values for the ELISA were observed among older cattle and cattle that aborted, compared with values for cattle with respiratory tract disease or other reasons for submission. CONCLUSIONS AND CLINICAL RELEVANCE: This laboratory-based surveillance sample provided a point estimate of seroprevalence against vesivirus among cattle in 9 US states. This suggests that vesivirus infection is widespread with high prevalence in some herds. Risk factors associated with vesivirus seroprevalence in beef and dairy cattle should be confirmed in population-based studies.

Evidence of Bovine viral diarrhea virus Infection in Three Species of Sympatric Wild Ungulates in Nevada: Life History Strategies May Maintain Endemic Infections in Wild Populations.

Evidence for bovine viral diarrhea virus (BVDV) infection was detected in 2009-2010 while investigating a pneumonia die-off in Rocky Mountain bighorn sheep (Ovis canadensis, canadensis), and sympatric mountain goats (Oreamnos americanum) in adjacent mountain ranges in Elko County, Nevada. Seroprevalence to BVDV-1 was 81% (N = 32) in the bighorns and 100% (N = 3) in the mountain goats. Serosurveillance from 2011 to 2015 of surviving bighorns and mountain goats as well as sympatric mule deer (Odocoileus hemionus), indicated a prevalence of 72% (N = 45), 45% (N = 51), and 51% (N = 342) respectively. All species had antibody titers to BVDV1 and BVDV2. BVDV1 was isolated in cell culture from three bighorn sheep and a mountain goat kid. BVDV2 was isolated from two mule deer. Six deer (N = 96) sampled in 2013 were positive for BVDV by antigen-capture ELISA on a single ear notch. Wild ungulates and cattle concurrently graze public and private lands in these two mountain ranges, thus providing potential for interspecies viral transmission. Like cattle, mule deer, mountain goats, and bighorn sheep can be infected with BVDV and can develop clinical disease including immunosuppression. Winter migration patterns that increase densities and species interaction during the first and second trimester of gestation may contribute to the long term maintenance of the virus in these wild ungulates. More studies are needed to determine the population level impacts of BVDV infection on these three species.

Infectious disease and the decline of Steller sea lions (Eumetopias jubatus) in Alaska, USA: insights from serologic data.

Serologic data were examined to determine whether infectious disease may have played a role in the decline of Steller sea lions (Eumetopias jubatus) in the Gulf of Alaska and Aleutian Islands, USA. Available published data, unpublished data, and recent collections (1997-2000) were compared and reviewed. Data were stratified by geography to compare the declining western Alaskan population in the Aleutian Islands through eastern Prince William Sound to the increasing population in southeastern Alaska. Prevalences of antibodies from the 1970s to the early 1990s were noted for Leptospira interrogans, Chlamydophila psittaci, Brucella spp., phocid herpesvirus-1, and calciviruses. Serum samples collected from 1997-2000 were tested for antibodies to these agents as well as to marine mammal morbilliviruses, canine parvovirus, and canine adenovirus-1 and -2. Conclusions could not be drawn about changes in antibody prevalence to these agents during the decline of Steller sea lions, however, because data were incomplete or not comparable as a result of inconsistencies in testing techniques. Despite these shortcomings, results provided no convincing evidence of significant exposure of Steller sea lions to morbilliviruses, Brucella spp., canine parvovirus, or L. interrogans. Steller sea lions have been exposed to phocid herpesviruses, caliciviruses, canine adenovirus, and C. psittaci or to cross-reactive organisms in regions of both increasing and decreasing sea lion abundance. Based on similar antibody prevalence estimates from the increasing and decreasing populations, these agents are unlikely to have been the primary cause of the population decline. They may have contributed to the decline or impeded population recovery, however, because of undetected mortality and morbidity or reductions of fecundity and body condition in animals under other stresses. Systematic monitoring for disease agents and their effects is needed to determine whether infectious disease currently plays a role in the decline and lack of recovery of Steller sea lions.

Serologic, trace element, and fecal parasite survey of free-ranging, female mule deer (Odocoileus hemionus) in eastern Washington, USA.

Blood and fecal samples collected from 97 free-ranging mule deer (Odocoileus hemionus), from four distinct herds during the spring of 2000 or 2001 in eastern Washington, US, were tested for exposure to selected pathogens, concentrations of trace elements, and presence of parasites in feces. Antibodies were detected to the following: Leptospira interrogans serovar Bratislava (4%), Leptospira interrogans serovar Canicola (1%), Leptospira interrogans serovar Grippotyphosa (13%), Bovine viral diarrhea virus 1 (57%), Bovine respiratory syncytial virus (71%), Infectious bovine rhinotracheitis virus (51%), Bovine parainfluenza virus 3 (61%), Bluetongue virus (25%), and Epizootic hemorrhagic disease virus (25%); 3 of 63 (5%) samples had antibody to Neospora spp. All samples tested for antibody to Brucella abortus and L. interrogans serovar Icterohaemorrhagiae, L. interrogans serovar Pomona, and L. interrogans serovar Hardjo samples were negative. Trace element concentrations from 97 sera were deficient for selenium (17%), copper (19%), iron (34%), calcium (3%), and phosphorus (2%) compared with thresholds established for domestic livestock. Parasites detected in 97 fecal samples included dorsal-spined larvae (probably Parelaphostrongylus sp.) (40%), abomasal nematode eggs (1%), Capillaria sp. eggs (1%), Nematodirus sp. eggs (26%), Moniezia sp. eggs (1%), and Eimeria sp. (2%).

Persistent Bovine Viral Diarrhea Virus Infection in Domestic and Wild Small Ruminants and Camelids Including the Mountain Goat (Oreamnos americanus).

Bovine viral diarrhea virus (BVDV) is a pestivirus best known for causing a variety of disease syndromes in cattle, including gastrointestinal disease, reproductive insufficiency, immunosuppression, mucosal disease, and hemorrhagic syndrome. The virus can be spread by transiently infected individuals and by persistently infected animals that may be asymptomatic while shedding large amounts of virus throughout their lifetime. BVDV has been reported in over 40 domestic and free-ranging species, and persistent infection has been described in eight of those species: white-tailed deer, mule deer, eland, mousedeer, mountain goats, alpacas, sheep, and domestic swine. This paper reviews the various aspects of BVDV transmission, disease syndromes, diagnosis, control, and prevention, as well as examines BVDV infection in domestic and wild small ruminants and camelids including mountain goats (Oreamnos americanus).

Abortion in a Mediterranean miniature donkey (Equus asinus) associated with a gammaherpesvirus similar to Equid herpesvirus 7.

Fetal tissues and placenta from a third trimester Mediterranean miniature donkey (Equus asinus) abortion were submitted to the Washington State University, Washington Animal Disease Diagnostic Laboratory for abortion diagnosis. Microscopic examination of formalin-fixed tissues revealed multifocal necrotizing placentitis. Several cells within the necrotic foci contained large, eosinophilic, intranuclear inclusions. Virus isolation from fresh, frozen placenta identified a cytopathic, syncytia-forming virus. Polymerase chain reaction (PCR) from the cultured virus using degenerate universal herpesvirus primers amplified a 699-base pair portion of the DNA polymerase gene. The PCR amplicon had 96.7% nucleotide identity with the DNA polymerase gene of Equid herpesvirus 7 (EHV-7; asinine herpesvirus 2), a gammaherpesvirus. An identical sequence was obtained when the same degenerate herpesvirus primers were used for PCR on the formalin-fixed placenta. Additionally, the amplicon had complete identity with short sequences of asinine herpesviruses that have been published in association with interstitial pneumonia in donkeys. EHV-7 has previously been isolated from nasal secretions of normal donkeys and mules. Our report describes a case of abortion associated with EHV-7 or a similar virus.

Bluetongue disease and seroprevalence in South American camelids from the northwestern region of the United States.

In late summer/early fall of 2013, 2 South American camelids from central Washington were diagnosed with fatal bluetongue viral disease, an event which is rarely reported. A 9-year-old intact male llama (Lama glama), with a 1-day history of anorexia, recumbency, and dyspnea before death. Abundant foam discharged from the mouth and nostrils, and the lungs were severely edematous on postmortem examination. Histologically, there was abundant intra-alveolar edema with fibrin. Hemorrhage and edema disrupted several other organs. Bluetongue viral RNA was detected by reverse transcription polymerase chain reaction (RT-PCR), and serotype 11 was identified by sequencing a segment of the VP2 outer capsid gene. Approximately 1 month later, at a site 150 miles north of the index case, a 2-year-old female alpaca with similar, acutely progressive clinical signs was reported. A postmortem examination was performed, and histologic lesions from the alpaca were similar to those of the llama, and again serotype 11 was detected by PCR. The occurrence of bluetongue viral infection and disease is described in the context of seasonal Bluetongue virus activity within the northwestern United States and southwestern Canada.

Clinical and epidemiologic observations of bovine viral diarrhea virus in the northwestern United States.

Retrospective analyses of cases from which bovine viral diarrhea virus (BVDV) was isolated from 1980 to 2000 were conducted. These cases originated from the northwestern US and included both beef and dairy cattle. The results indicated that there was a shift in diseases associated with BVDV infection and in the animal age at onset of disease. Comparative results from the 1980 data indicated a low fetal infection rate (<5%), followed by steady increases of clinical cases and peaking at 6 months (30%). By 2000, the shift of BVDV cases was noticeable and indicated a biphasic occurrence of disease. The first phase was fetal infections, which increased to >25%, followed by a second phase at 6 months (>35%). Phylogenetic analysis was conducted on selected isolates from the time period 1998-2000 (n = 54). There were representative viral isolates from the two genotypes (BVDV1 and BVDV2), as well as subgenotypes, BVDV1a and BVDV1b. The types were further correlated with the clinical manifestation, which were reported as mucosal disease, persistently infected (PI)-poor doer, and abortion-open cows. The results indicated that BVDV were distributed throughout the clinical spectrum of disease, with BVDV2 representing the greatest frequency of isolation, and the greatest association with abortion-open cows. When the BVDV genotypes and subgenotypes were categorized into early (<100 days gestation) versus late (>100 days gestation) fetal infections, there was an inverse relationship noted. It was observed that BVDV1a was associated least with early infection (14%) and most with late infections (86%). BVDV1b was intermediate, followed by BVDV2, which was associated more with early infections (45%) and less with late infections (55%) when compared with BVDV1a and BVDV1b.

Results of a new serologic test suggest an association of Waddlia chondrophila with bovine abortion.

Waddlia chondrophila is a little-known intracellular organism belonging to the order Chlamydiales that has twice been isolated from aborted bovine fetuses. To initiate an investigation of the possibility that W. chondrophila may be an abortifacient pathogen, a serologic test was developed and used to screen bovine sera that were submitted to the Washington Animal Disease Diagnostic Laboratory (Pullman, WA). A highly significant statistical association (P < 0.00001) was observed when comparing antibody titers in cows that had aborted with those in other classes of cattle. Although this result is consistent with the possibility that infection with W. chondrophila may be a cause of bovine abortion, it is also possible that seroprevalence simply increases with age or that exposure rates differ between cows and other classes of cattle. Future serologic studies should specifically compare antibody titers between aborting cows and carefully matched nonaborting cohorts.

Comparison of targeting F and G protein genes to detect bovine and ovine respiratory syncytial viruses.

In this study, 2 reverse transcriptase-polymerase chain reaction (RT-PCR) assays were developed and compared for simultaneous detection of bovine and ovine respiratory syncytial viruses (RSVs). One assay was based on a set of primers, which amplified a 426-bp fragment of either bovine or ovine RSV F gene (RT-PCR F). The F products could be distinguished by EcoRI or BstYI restriction endonuclease cleavage. In the other assay, a set of primers amplified a 542-bp fragment of either ovine or bovine RSV G gene (RT-PCR G). EcoO1091 and RsaI restriction enzymes were used to differentiate between the ovine and bovine PCR-G products. Sequencing of the PCR products confirmed the fidelity of both assays. The 2 assays were evaluated using 18 bovine RSV isolates, 1 ovine RSV, 1 bighorn sheep RSV isolate, 1 caprine RSV isolate, 2 human RSV isolates, and several other viruses associated with bovine respiratory tract disease. RT-PCR G may be more sensitive in detecting viral RNA. Because the target sequence of the F gene is more conserved than that of the G gene, RT-PCR F followed by the appropriate restriction enzyme cleavage may be superior to RT-PCR G to discriminate between the 2 ruminant RSV subgroups. This assay should prove useful for determining the relative contribution of ovine and bovine RSV to the pathogenesis of bovine respiratory tract disease.

Temporal patterns of diagnostic results in serial samples from cattle with advanced paratuberculosis infections.

As part of investigating diagnostic strategies for Mycobacterium avium subsp. paratuberculosis (Map), serial results from polymerase chain reaction (PCR) on extraintestinal tissues (blood, milk, and liver) were compared with those from more conventional detection methods including serum enzyme-linked immunosorbent assay (ELISA), fecal culture, and fecal PCR. Three cows previously identified as being subclinically infected with Map were selected for the study. Blood, milk, and feces were collected daily and liver biopsies were obtained weekly for a 30-day period. Unexpectedly, a substantial daily variation in serum ELISA sample to positive (S/P) ratios was observed in all 3 cows. In contrast, fecal culture results were consistently positive. However, whereas fecal culture colony counts were consistently high for 2 cows throughout the study, colony counts from the third cow varied from day to day. Diagnostic sensitivity of PCR for fecal, blood, milk, and liver samples in these advanced subclinically infected cows was 87%, 40%, 96%, and 93%, respectively.

Serosurvey of viral infections in free-ranging Namibian cheetahs (Acinonyx jubatus).

Cheetahs (Acinonyx jubatus) in captivity have unusually high morbidity and mortality from infectious diseases, a trait that could be an outcome of population homogeneity or the immunomodulating effects of chronic stress. Free-ranging Namibian cheetahs share ancestry with captive cheetahs, but their susceptibility to infectious diseases has not been investigated. The largest remaining population of free-ranging cheetahs resides on Namibian farmlands, where they share habitat with domestic dogs and cats known to carry viruses that affect cheetah health. To assess the extent to which free-ranging cheetahs are exposed to feline and canine viruses, sera from 81 free-ranging cheetahs sampled between 1992 and 1998 were evaluated for antibodies against canine distemper virus (CDV), feline coronavirus (feline infectious peritonitis virus; FCoV/ FIPV), feline herpesvirus 1 (FHV1), feline panleukopenia virus (FPV), feline immunodeficiency virus (FIV), and feline calicivirus (FCV) and for feline leukemia virus (FeLV) antigens. Antibodies against CDV, FCoV/FIPV, FHV1, FPV, and FCV were detected in 24, 29, 12, 48, and 65% of the free-ranging population, respectively, although no evidence of viral disease was present in any animal at the time of sample collection. Neither FIV antibodies nor FeLV antigens were present in any free-ranging cheetah tested. Temporal variation in FCoV/FIPV seroprevalence during the study period suggested that this virus is not endemic in the free-ranging population. Antibodies against CDV were detected in cheetahs of all ages sampled between 1995 and 1998, suggesting the occurrence of an epidemic in Namibia during the time when CDV swept through other parts of sub-Saharan Africa. This evidence in free-ranging Namibian cheetahs of exposure to viruses that cause severe disease in captive cheetahs should direct future guidelines for translocations, including quarantine of seropositive cheetahs and preventing contact between cheetahs and domestic pets.

Biosecurity for neonatal gastrointestinal diseases.

Infectious diarrhea is an important cause of neonatal calf morbidity and mortality that results in significant economic losses in the beef and dairy industries. Although numerous risk factors related to the occurrence of neonatal diarrhea have been identified, they can all be categorized into those that are related to the calf, the pathogens involved, or the environment of the calf. The immune status of calves, specifically the level of passively acquired immunity through colostrum, is the major risk factor related to the calf and the occurrence of diarrhea. Although numerous pathogens have been implicated in the occurrence of neonatal diarrhea, only a relatively limited number are commonly involved. Most should be viewed as secondary opportunists rather than primary pathogens, because none are extraordinarily virulent, and with the exception of Salmonella spp., most are present within the gastrointestinal tract of many healthy, mature cattle. Important risk factors related to pathogens involved in neonatal calf diarrhea involve the size of the inoculum and the occurrence of multiple infections. Finally, when considering the environment and housing conditions in which beef and dairy calves may reside, it is clear that tremendous variations exist. Despite these variations, the risk factors associated with the environment of the calf are also those that are the most amenable to the implementation of general environmental control and monitoring strategies as well as specific biosecurity measures.