Serological evidence for exposure to bovine viral diarrhoea virus in sheep co-grazed with beef cattle in New Zealand

ABSTRACT

Aims: To determine whether sheep that co-grazed with cattle that were suspected to be positive for bovine viral diarrhoea (BVD) virus had serological evidence of exposure to the virus.

Methods: Eighteen commercial farms that routinely co-grazed cattle and sheep in the same paddocks were recruited through purposive sampling. The recruiting veterinarians identified nine farms with cattle herds that were known or highly suspected to be positive for BVD and nine farms that were considered to be free of BVD. Blood samples were taken from 15 ewes aged 1 year on each farm and samples were submitted to a commercial diagnostic laboratory to test for antibodies against pestiviruses using an ELISA. All samples that were positive were then tested using a virus neutralisation test (VNT)for antibodies against BVD virus.

Results: Of the 270 blood samples, 17 were positive for pestivirus antibodies by ELISA and these originated from two farms that were known or suspected to have BVD virus-positive cattle. None of the samples from the nine flocks co-grazed with cattle herds that were known or suspected to be BVD virus-negative were positive for pestivirus antibodies. Within the two positive farms, 2/15 samples from the first farm and 15/15 samples from the second farm were antibody-positive. When the 17 positive blood samples were submitted for VNT, all 15 samples from the second farm tested positive for BVD virus antibodies with the highest titre being 1:512.

Conclusions and clinical relevance: In this small sample of New Zealand sheep and beef farms with suspected BVD infection in cattle, there was evidence of pestivirus exposure in co-grazed sheep. Although we were unable to confirm the origin of the exposure in these sheep, these findings highlight that farmers who are trying to eradicate BVD from their cattle should be mindful that the infection may also be circulating in sheep, and both populations should be considered a possible risk to each other for generating transient and persistent infections. Further work is needed to estimate the true prevalence of New Zealand sheep flocks that are affected by BVD and the associated economic impacts.     

Confirmation of the occurrence of the chewing louse Bovicola (Lepikentron) breviceps (Insecta: Phthiraptera: Trichodectidae) on alpacas (Lama pacos) in New Zealand

Abstract

AIM: To investigate the cause of classical swine fever (CSF) virus-seropositive animals in a nucleus pig-breeding herd in New Zealand, where porcine circovirus-associated disease had been diagnosed.

CASE HISTORY AND CLINICAL FINDINGS: An exotic disease investigation was undertaken to exclude CSF and porcine reproductive and respiratory syndrome (PRRS) on a nucleus pig-breeding herd comprising approximately 300 breeding sows, 1,000 weaners, and 650 grower pigs. The herd was experiencing poor reproductive performance in sows, and breeding records showed a declining farrowing rate attributable to a single manager. The growing pigs (10–15 weeks old) were experiencing respiratory disease and wasting, and the mortality rate by pen varied between 9 and 20%. Post-mortem changes in affected grower pigs were consistent with circovirus-associated diseases.

DIAGNOSTIC TESTING: Serological screening using an IDEXX-ELISA gave negative results for PRRS virus antibodies, but two grower pigs and one sow tested positive for CSF virus antibodies. These three seropositive animals remained positive to CSF virus, using three commercial ELISA test kits, over 27 weeks. A newly developed virus neutralisation test (VNT), using a New Zealand isolate of border disease (BD) virus, demonstrated that the seropositive pig sera had higher antibody titres to BD virus than to bovine viral diarrhoea (BVD) virus and CSF virus.

PCR performed on tonsil, kidney, ileum and spleen gave negative results for CSF virus, and histopathology on lymph nodes, intestine, lung, kidney, liver and brain showed no evidence of the disease. Virus isolation performed on a number of samples was negative.

CLINICAL RELEVANCE: The seropositive samples for CSF virus found in this investigation were likely to be a cross reaction to a pestivirus other than CSF virus. The finding of a possible endemic pestivirus capable of being transmitted between sheep and pigs on this farm may explain findings from previous serological survey work in New Zealand, and supports experience elsewhere, where BD virus was found to be the predominant ruminant pestivirus infecting pigs. The results show that pestivirus cross reactivity can result in unexpectedly high titres, and that testing with a full set of (local) pestiviruses is necessary to reach the correct conclusion. The investigation has direct relevance where pig herds with a low seroprevalence are encountered during surveillance for CSF.     

Development of an indirect ELISA for detection of antibody to wobbly possum disease virus in archival sera of Australian brushtail possums (Trichosurus vulpecula) in New Zealand

AIMS: To develop an indirect ELISA based on recombinant nucleocapsid (rN) protein of wobbly possum disease (WPD) virus for investigation of the presence of WPD virus in Australian brushtail possums (Trichosurus vulpecula) in New Zealand.

METHODS: Pre- and post-infection sera (n=15 and 16, respectively) obtained from a previous experimental challenge study were used for ELISA development. Sera were characterised as positive or negative for antibody to WPD virus based on western-blot using WPD virus rN protein as antigen. An additional 215 archival serum samples, collected between 2000–2016 from five different regions of New Zealand, were also tested using the ELISA. Bayesian modelling of corrected optical density at 450?nm (OD₄₅₀) results from the ELISA was used to obtain estimates of receiver operating characteristic (ROC) curves to establish cut-off values for the ELISA, and to estimate the prevalence of antibody to WPD virus.

RESULTS: Western blot analysis showed 5/14 (36%) pre-infection sera and 11/11 (100%) post-infection sera from experimentally infected possums were positive for antibodies to WPD virus. Bayesian estimates of the ROC curves established cut-off values of OD₄₅₀≥0.41 for samples positive, and OD₄₅₀<0.28 for samples negative for antibody to WPD virus, for sera diluted 1:100 for the ELISA. Based on the model, the estimated proportion of samples with antibodies to WPD virus was 0.30 (95% probability interval=0.196–0.418). Of the 230 archival serum samples tested using the ELISA, 48 (20.9%) were positive for antibody to WPD virus, 155 (67.4%) were negative and 27 (11.7%) equivocal, using the established cut-off values. The proportion of samples positive for WPD virus antibody differed between geographical regions (p<0.001).

CONCLUSION: The results suggested that WPD virus or a related virus has circulated among possums in New Zealand with differences in the proportion of antibody-positive samples from different geographical regions. Antibodies to WPD virus did not seem to protect possums from disease following experimental infection, as one third of possums from the previous challenge study showed evidence of pre-existing antibody at the time of challenge. These results provide further support for existence of different pathotypes of WPD virus, but the exact determinants of protection against WPD and epidemiology of infection in various regions of New Zealand remain to be established.

CLINICAL RELEVANCE: Availability of the indirect ELISA for detection of WPD virus antibody will facilitate prospective epidemiological investigation of WPD virus circulation in wild possum populations in New Zealand.     

Prevalence of Neospora antibodies in beef cattle in New Zealand

dAim:To estimate the prevalence of Neospora infection in a sample of New Zealand beef cattle.

dMethods: The prevalence of Neospora caninum infection in New Zealand beef cattle was estimated by collecting blood at slaughter from 499 beef cattle from 40 different farms at 2 slaughter plants in the North Island and 1 in the lower South Island. Sera were tested using an ELISA against Neospora tachyzoite antigen.

dResults: The prevalence of seropositive cattle was 2.5% (n=120), 3.6% (n=166) and 2.3% (n=213) at the plants surveyed, the overall prevalence being 2.8%. The serologically positive cattle came from 9 farms, 3 of which had more than 1 positive animal. The highest prevalence recorded amongst animals from 1 farm was 4/13 (31%), in a group of young steers.

dConclusion: Neosporosis appears to be present at a lower level in the New Zealand beef cattle population than in the New Zealand dairy cattle population. Nevertheless, from the high seroprevalence evident amongst young cattle on 1 farm, we suggest that Neospora may be a cause of infertility in beef cattle in this country.     

Detection of bovine herpesvirus type 4 antibodies and bovine lymphotropic herpesvirus in New Zealand dairy cows

AIM: To detect the presence of bovine herpesvirus (BoHV) type 4 in New Zealand dairy cows with clinical metritis.

METHODS: Serum samples taken from 92 dairy cows with clinical metritis, each from a different farm, were tested for the presence of antibodies against BoHV-4 using a commercially available, indirect ELISA. Peripheral blood mononuclear cells (PBMC) were collected from 10 BoHV-4 seropositive cows, and PBMC were examined by a pan-herpesvirus nested PCR to detect herpesvirus. PCR products were sequenced directly and a proportion of the PCR products were cloned and sequenced to identify the virus present.

RESULTS: Antibodies to BoHV-4 were detected in 23/92 (25%) serum samples. The pan-herpesvirus PCR was positive in 8/10 PBMC samples. Cloning and sequencing identified that all of the eight PCR-positive PBMC contained bovine lymphotropic herpesvirus (BLHV); no BoHV-4 DNA was detected.

CONCLUSIONS: This study reports the finding of the presence of apparent antibodies to BoHV-4, and BLHV DNA in New Zealand dairy cows affected by metritis.

CLINICAL RELEVANCE: Bovine herpesvirus type 4 and BLHV are reported to have the potential to cause reproduction failure in cows. This is the first report of apparent BoHV-4 antibodies, and BLHV in New Zealand. The importance and epidemiology of these viruses in cattle in New Zealand requires further investigation.     

Prevalence of antibodies to Neospora caninum in dogs of rural or urban origin in central New Zealand

AIM: To investigate the seroprevalence of Neospora caninum infection in populations of dogs from dairy farms, sheep/beef farms and urban areas in the central part of New Zealand. It was postulated seroprevalence would be higher for farm dogs than urban dogs if the life-cycle of this parasite involves transmission between dogs and cattle.

METHODS: Serum samples were obtained from dogs that lived on dairy farms (n=161), sheep/beef farms (n=154) and in urban situations (n=150). The relative risk of detecting antibodies to N. caninum using an immunofluorescent antibody test (IFAT) was compared between farm and urban dogs.

RESULTS: The relative risk of having a titre of ≥1:200 to N. caninum was 2.43 (95% CI=1.88-3.14) for dairy-farm dogs and 3.16 (95% CI=2.48–4.02) for sheep/beef-farm dogs, compared with urban dogs. At this titre, which is currently used in New Zealand to indicate seropositivity, seroprevalence of N. caninum infection was 30.7% in urban dogs, 74.5% in dairy-farm dogs and 96.8% in sheep/beef-farm dogs.

CONCLUSION: This observation is consistent with a cycling of this disease between cattle and dogs on farms in New Zealand and with higher exposure of dogs to N. caninum on farms than occurs in urban environments. The prevalence of antibodies in all three groups of dogs tested in this study (dairy-farm dogs, sheep/beef-farm dogs and urban dogs) is higher than has generally been reported elsewhere. New Zealand farm dogs have a higher serological prevalence of N. caninum infection than urban dogs.

CLINICAL RELEVANCE: Management and disease control practices that break the life-cycle of transmission between cattle and dogs should assist in controlling cattle abortion due to N. caninum.     

Comparison of the Q-fever complement fixation test and two commercial enzyme-linked immunosorbent assays for the detection of serum antibodies against Coxiella burnetii (Q-fever) in ruminants: Recommendations for use of serological tests on imported animals in New Zealand

Abstract

AIM: To make valid recommendations on the use of serological test methods for the detection of serum antibodies in ruminants against Coxiella burnetii (Q-fever), by comparing the performance of the complement fixation test (CFT) and two ELISA, and by identifying reasons for discrepancies between the test methods.

METHODS: A total of 73 serum samples from infected cattle, 69 from infected goats, and 100 samples from non-infected cattle and 57 samples from non-infected sheep, as well as 95 samples from infected cattle herds (mix of seropositive and seronegative samples), were tested using the CFT, the IDEXX ELISA (I-ELISA) and the Pourquier ELISA (P-ELISA). A mixed panel of 12 serum samples from sheep from inter-laboratory proficiency testing (proficiency panel) was also tested using the CFT and both ELISA, and further investigated using IgG- and IgM-specific ELISA.

RESULTS: Generally, the two commercial ELISA were more sensitive than the CFT for the detection of infected ruminants. Good agreement between ELISA for positive and negative results was found for samples from the infected herd, while results for the positive panels varied between the two ELISA. For the total of the positive serum panels, the I-ELISA detected 95% of samples as positive or suspicious, while the P-ELISA detected only 81%. In the P-ELISA, more samples were considered suspicious (18%) than in the I-ELISA (14%). All sera from noninfected sheep and cattle tested negative in the serological test methods employed, except for one positive sample from a sheep in the P-ELISA. Further investigation revealed that a CFT-positive but ELISA-negative result was due to high IgM and low IgG reactivity.

CONCLUSIONS: The two commercial ELISA were more sensitive than the CFT in all panels from infected ruminants. However, they could only detect IgG. The I-ELISA should be the serological test method of choice for cattle, sheep and goats for import testing of animals into New Zealand because it was more sensitive than the P-ELISA and was equally specific to the PELISA and the CFT. For other animal species, such as deer and camelids, the CFT should still be used since none of the ELISA has been evaluated for these species. This study has shown that the two commercial ELISA will detect the majority of infected ruminants but may miss animals that have not developed an IgG response.     

Seroprevalence of Toxoplasma gondii in mainland and sub-Antarctic New Zealand sea lion (Phocarctos hookeri) populations

AIMS: To investigate the seroprevalence of antibodies to Toxoplasma gondii in New Zealand sea lions (Phocarctos hookeri), as a potential contributor to reproductive failure.

METHODS: Archived sera were sourced from New Zealand sea lions from two recolonising mainland populations in the Otago Peninsula (n=15) and Stewart Island (n=12), as well as a declining population at Enderby Island (n=28) in the New Zealand sub-Antarctic. Sera were tested for antibodies to T. gondii using a commercially available ELISA (with samples considered positive if the sample to positive ratio was?>30%), and latex agglutination test (LAT; with titres ≥1:32 considered positive). Western blot analysis was used to validate the results of a subset of 14 samples.

RESULTS: Five samples from sea lions in mainland locations were confirmed positive for antibodies to T. gondii. Two adult females exhibited high LAT antibody titres (min 1:2048, max 1:4096) on both occasions when sampled 1 and 2 years apart, respectively. No animals from Enderby Island were seropositive.

CONCLUSIONS: Toxoplasma gondii infection is unlikely to be a major contributor to poor reproductive success in New Zealand sea lions. However, continued surveillance is pertinent to assess subclinical and clinical impacts of the parasite on these threatened populations. The commercial tests evaluated here, with further species-specific threshold refinement could provide a fast, inexpensive and reliable indicator of T. gondii exposure in New Zealand sea lions.     

Practices and opinions of New Zealand beef cattle farmers towards bovine viral diarrhoea control in relation to real and perceived herd serological status

ABSTRACT

Aims: To investigate the seroprevalence of infection with bovine viral diarrhoea (BVD) virus among 75 beef herds and seroconversion in cattle during early pregnancy, and to determine the practices and opinions of farmers towards BVD control and their association with real and perceived herd serological status.

Methods: Blood samples were collected before mating in 75 beef herds across New Zealand from 15 unvaccinated heifers that had delivered their first calf that season. Serum samples were tested for BVD antibodies using ELISA individually, and after pooling samples for each farm. Animals that were antibody-negative were retested at either pregnancy diagnosis or weaning. Farmers were asked to complete a detailed survey about herd demographics, BVD testing and vaccination practices, and opinions towards national BVD control.

Results: Based on the pooled serum antibody ELISA results, there were 28/75 (37%) negative herds, 15/75 (20%) suspect herds, and 32/75 (43%) positive herds. Of 1,117 animals sampled 729 (65.3%) tested negative for BVD virus antibodies; when retested, 47/589 (8.0%) animals from 13/55 (24%) herds had seroconverted. Among 71 famers providing survey responses 11 (15%) believed their herd was infected with BVD, 24 (34%) were unsure and 36 (51%) did not think their herd was infected. Only 19/71 (18%) farmers had performed any BVD testing within the past 5 years and 50/70 (71%) had not vaccinated any cattle for BVD. Support for national BVD eradication programme was strong in 51/71 (56%) respondents, but the biggest challenge to BVD control was considered to be famer compliance. Compared to farmers who did not think their herd was infected, more farmers who thought BVD was present in their herds had previously tested for BVD, would consider testing all replacement calves, and would support establishing a national BVD database; fewer would consider purchasing BVD tested or vaccinated cattle only.

Conclusions and clinical relevance: Only 15% of the beef farmers in this study believed their herds were infected with BVD virus and few of them had undertaken BVD screening. Nevertheless many were supportive of implementing a national BVD control programme. It is likely that the lack of farmer awareness around BVD and the failure of farmers to recognise the potential impacts in their herds are hindering progress in controlling the disease in New Zealand. There are opportunities for New Zealand veterinarians to be more proactive in helping beef farmers explore BVD management options.     

Validation of a bulk tank milk antibody ELISA to detect dairy herds likely infected with bovine viral diarrhoea virus in New Zealand

AIMS: To assess the sensitivity and specificity of a bulk tank milk (BTM) antibody enzyme-linked immunosorbent assay (ELISA) to detect likely infection of a dairy herd with bovine vi- ral diarrhoea virus (BVDV). The ELISA was subsequently used to estimate the prevalence of likely infected herds in parts of the North Island of New Zealand.

METHODS: BTM samples from 724 randomly selected dairy herds in the Waikato, Bay of Plenty and Northland regions of New Zealand were tested for BVDV antibodies. From this group, 20 herds were again randomly selected from each of the quartiles of the ELISA percentage inhibition (%INH) result. From each participant herd, serum from 15 randomly selected calves aged 6–18 months and 15 cows was collected and tested using an indirect blocking ELISA for BVDV antibodies.

RESULTS: Among serum results from calves from 50 herds available for analysis, 34 (68%) herds were classified as likely non-infected (0-3 seropositive among 15 calves) and 16 (32%) as likely infected (5–15 seropositive among 15 calves). Receiver- operator characteristic (ROC) analysis identified an optimal cut-off for BTM of 80%INH associated with 81% sensitivity and 91% specificity for likely herd infection. The prevalence of BVDV antibodies in cows within herds and %INH for BVDV in bulk milk were positively correlated (p<0.01). The association between bulk milk %INH and the prevalence of BVDV antibodies in calves was stronger than the same association in cows. Based on the threshold of 80%INH, the 95% confidence interval (CI) for prevalence of likely infection in the 724 herds in the Waikato, Bay of Plenty and Northland regions of New Zealand was 12–17%. Vaccination against BVDV was not significantly associated with the likely infection status of the herd based on prevalence of BVDV antibodies among calves.

CONCLUSION: An ELISA test result for BVDV antibodies in BTM ≥80%INH can be used as a threshold to indicate the presence of likely infection with BVDV in dairy herds in New Zealand, with 81% sensitivity and 91% specificity.     

Prevalence of Candidatus Mycoplasma haemolamae,bovine viral diarrhoea virus,and gastrointestinal parasitism in a sample of adult New Zealand alpaca (Vicugna pacos)

AIM: To determine the prevalence of infection with Candidatus Mycoplasma haemolamae (Mhl), antibodies to bovine viral diarrhoea virus (BVDV), and BVDV antigen, and the prevalence of animals with elevated faecal nematode egg counts (FEC) in a sample of adult New Zealand alpaca (Vicugna pacos).

METHODS: Blood samples were obtained from 175 alpaca, collected from 15 farms around New Zealand, and from 31 samples sent to a diagnostic laboratory for routine haematology. Blood smears (n=170) were examined microscopically for the presence of haemoplasma, and DNA was extracted from whole blood (n=206) for real-time PCR testing for Mhl. Packed cell volume (PCV) was determined for 193 samples. Serum samples (n=195) were tested for BVDV antibody using ELISA, and for BVDV antigen using a real-time PCR assay. Faecal samples were collected from 143 animals; FEC were measured, and samples pooled for larval culture.

RESULTS: No haemoplasma organisms were present on blood smear examination. Of the 206 blood samples, two (from the same farm) were positive for Mhl by real-time PCR testing, giving a prevalence of infection with Mhl of 0.97%. Of the 195 serum samples tested, four (2.1%) were positive for antibodies to BVDV; animals with BVDV antibodies were from 3/15 (20%) farms, none of which farmed cattle. None of the serum samples were positive by PCR for BVDV antigen. The median FEC was 50?epg (min 0, max 4,700), with 55/143 (38.5%) samples having 0?epg, and 33/143 (23.1%) having ≥250?epg. Haemonchus spp. were the most common nematodes present in faecal larval cultures from the North Island. Log₁₀ FEC was negatively associated with PCV (p=0.02), and was higher in males than females (p<0.001), and in animals that were positive compared with negative for Mhl (p=0.022).

CONCLUSIONS AND CLINICAL RELEVANCE: The number of alpaca infected with Mhl was low, as was the seroprevalence of BVDV. Gastrointestinal parasitism was, however, a common finding in this sample of New Zealand alpaca.     

Feline leukaemia virus testing

Extract

Sir:- In September 1981 we published the results of leukaemia virus (Felv) testing of cats in New Zealand.⁽²⁾ In addition we detailed recommendations for Felv testing of cats in this country. These recommendations were revised and updated in a second letter to the Journal in 1982.⁽³⁾ Since then a further 465 cats have been tested at Massey University using the leukassay test (ELISA) (Leukassay F, Pitman Moore).     

The seroprevalence of canine respiratory coronavirus and canine influenza virus in dogs in New Zealand

Abstract

AIM: To determine whether canine respiratory coronavirus (CRCoV) and canine influenza virus (CIV) are present in dogs in New Zealand.

METHODS: Serum samples from 251 dogs of varying age, breed and clinical histories were tested for the presence of antibodies to CRCoV and CIV, using indirect fluorescent antibody (IFA) analysis. The population sampled represented a wide geographic area but principally encompassed the central and lower North Island of New Zealand.

RESULTS: Seventy-three of the 251 samples (29%) were seropositive for CRCoV. Dogs <2 years old were less likely to be seropositive for CRCoV than older dogs. None was seropositive for CIV.

CONCLUSIONS: This study revealed the presence of antibodies to CRCoV in dogs in New Zealand. Young dogs are less likely to be seropositive than older dogs, probably due to increased opportunity for exposure to CRCoV over time. Serum antibodies to CIV were not detected in any of the dogs sampled, suggesting that this virus is unlikely to be present in dogs in New Zealand.

CLINICAL RELEVENCE: Canine respiratory coronavirus is present in New Zealand. Although the role of this virus in canine infectious tracheobronchitis has not been fully elucidated, evidence suggests that it may have a causal role in this disease. Veterinarians should consider CRCoV as a differential diagnosis in cases of respiratory disease in dogs in New Zealand. While CIV appears not to be currently present in New Zealand, veterinarians should consider infection with this virus as a differential diagnosis in dogs presenting with respiratory signs.     

Prevalence of antibodies to infectious bovine rhinotracheitis, parainfluenza 3, bovine respiratory syncytial, and bovine viral diarrhea viruses in cattle in Saskatchewan and Alberta

A total of 1745 healthy cattle from 295 farms in Saskatchewan and Alberta was tested by ELISA for antibodies to four viruses. Antibodies to infectious bovine rhinotracheitis (IBR) virus were found in 37.8% of sera (59.5% of properties), to parainfluenza 3 (PI3) virus in 93.9% of sera (99.7% of properties), to bovine respiratory syncytial (BRS) virus in 78.5% of sera (86.6% of properties), and to bovine viral diarrhea (BVD) virus in 40.6% of sera (66.7% of properties)

The prevalence of PI3 viral antibodies among Saskatchewan cattle was not affected by district of origin, breed, sex, age, or vaccination practices, though BRS viral antibodies appeared less frequent in young, male, and unvaccinated animals. Antibodies to IBR and BVD viruses were less prevalent in the Prince Albert/Tisdale districts and in young, male, and unvaccinated animals, but were more common in Holstein cattle. Antibodies to IBR virus appeared less frequent in Herefords. Antibodies were more prevalent in cattle which had been vaccinated against IBR, BRS, and BVD virus infections.

The relatively small number of cattle sampled from Alberta had a similar prevalence of antibodies to PI3 and BRS viruses to that seen in cattle in Saskatchewan, though IBR and BVD prevalence rates were lower.

     

Use of an enzyme-linked immunosorbent assay for the detection of IgG antibodies to rinderpest virus in epidemiological surveys

A comparison was made between an indirect enzyme-linked immunosorbent assay (ELISA) and the virus neutralisation test (VNT) for the detection of antibodies to rinderpest virus in field sera. The results did not agree for 6 per cent of the sera tested where 3 per cent of the samples gave ELISA positive/VNT negative and 3 per cent gave ELISA negative/VNT positive. The latter sera all had high levels of IgM antibody, which may indicate animals being at an early stage of infection or detection of a non-specific reaction. The ELISA results give a representative picture of the immune status for field surveys and a greater number of sera can be assayed with relative ease, compared to the traditional serum neutralisation test.     

A competitive ELISA using anti-N monoclonal antibodies for specific detection of rinderpest antibodies in cattle and small ruminants.

A competitive ELISA (C-ELISA) using monoclonal antibodies (mAbs) which bind to the nucleo-protein (NP) of rinderpest virus (RPV) for detection of RPV antibodies in cattle and small ruminant sera is described. Unlike virus neutralisation test (VNT), this test using mAb IVB2-4, can detect specific RPV antibodies without showing a cross-reaction with antibodies to peste-des-petits ruminants-virus (PPRV); by contrast, when mAb VE4-1 is used the test detects both RPV and PPRV antibodies, including low levels of antibodies that can be found in sera containing maternal antibodies. Although antibodies to the PPRV 75-1 strain are also detected with mAb 51-5-6, the test is suitable for assessing the immune status of cattle against the Rinderpest Old Kabete (RBOK) strain. The results from a panel of sera with a known status of vaccination provide evidence for a highly significant correlation between C-ELISA and VNT. This test may be a useful tool for a standardized and accurate determination of the immunity status of both cattle and small ruminants.     

Viruses associated with outbreaks of equine respiratory disease in New Zealand

AIM: To identify viruses associated with respiratory disease in young horses in New Zealand.

METHODS: Nasal swabs and blood samples were collected from 45 foals or horses from five separate outbreaks of respiratory disease that occurred in New Zealand in 1996, and from 37 yearlings at the time of the annual yearling sales in January that same year. Virus isolation from nasal swabs and peripheral blood leukocytes (PBL) was undertaken and serum samples were tested for antibodies against equine herpesviruses (EHV-1, EHV-2, EHV-4 and EHV-5), equine rhinitis-A virus (ERAV), equine rhinitis-B virus (ERBV), equine adenovirus 1 (EAdV-1), equine arteritis virus (EAV), reovirus 3 and parainfluenza virus type 3 (PIV3).

RESULTS: Viruses were isolated from 24/94 (26%) nasal swab samples and from 77/80 (96%) PBL samples collected from both healthy horses and horses showing clinical signs of respiratory disease. All isolates were identified as EHV-2, EHV-4, EHV-5 or untyped EHV. Of the horses and foals tested, 59/82 (72%) were positive for EHV-1 and/or EHV-4 serum neutralising (SN) antibody on at least one sampling occasion, 52/82 (63%) for EHV-1-specific antibody tested by enzyme-linked immunosorbent assay (ELISA), 10/80 (13%) for ERAV SN antibody, 60/80 (75%) for ERBV SN antibody, and 42/80 (53%) for haemagglutination inhibition (HI) antibody to EAdV-1. None of the 64 serum samples tested were positive for antibodies to EAV, reovirus 3 or PIV3. Evidence of infection with all viruses tested was detected in both healthy horses and in horses showing clinical signs of respiratory disease. Recent EHV-2 infection was associated with the development of signs of respiratory disease among yearlings [relative risk (RR)=2.67, 95% CI=1.59-4.47, p=0.017].

CONCLUSIONS: Of the equine respiratory viruses detected in horses in New Zealand during this study, EHV-2 was most likely to be associated with respiratory disease. However, factors other than viral infection are probably important in the development of clinical signs of disease.     

Mixing of selenium and anthelmintic drench

Extract

Bovine viral diarrhoea (BVD) virus has a worldwide distribution and investigations in various parts of the world have shown that 60%–80% of cattle have neutralising antibodies to the virus⁽¹⁾⁽²⁾. Bovine viral diarrhoea virus infection is also common in New Zealand dairy herds⁽³⁾, and its epidemiology on dairy farms is well understood. It had been considered that the traditional beef cattle population was essentially free from this infection and there was a concern that the rapidly expanding dairy-beef industry may introduce infection into an essentially naive beef cattle population. However, a recent study has shown that BVD virus infection is widespread in beef herds throughout New Zealand⁽⁴⁾. To explore the issue further, we have examined the prevalence of BVD virus antibody- positive animals in selected dairy-beef operations and traditional cow-calf herds, and how BVD-virus infection, if present, is maintained within these cattle populations.     

Infection with Menangle virus in flying foxes (Pteropus spp.) in Australia

Objective To examine flying foxes (Pteropus spp.) for evidence of infection with Menangle virus. Design Clustered non‐random sampling for serology, virus isolation and electron microscopy (EM). Procedure Serum samples were collected from 306 Pteropus spp. in northern and eastern Australia and tested for antibodies against Menangle virus (MenV) using a virus neutralisation test (VNT). Virus isolation was attempted from tissues and faeces collected from 215 Pteropus spp. in New South Wales. Faecal samples from 68 individual Pteropus spp. and four pools of faeces were examined by transmission EM following routine negative staining and immunogold labelling. Results Neutralising antibodies (VNT titres ≥ 8) against MenV were detected in 46% of black flying foxes (P. alecto), 41% of grey‐headed flying foxes (P. poliocephalus), 25% of spectacled flying foxes (P. conspicillatus) and 1% of little red flying foxes (P. scapulatus) in Australia. Positive sera included samples collected from P. poliocephalus in a colony adjacent to a piggery that had experienced reproductive disease caused by MenV. Virus‐like particles were observed by EM in faeces from Pteropus spp. and reactivity was detected in pooled faeces and urine by immunogold EM using sera from sows that had been exposed to MenV. Attempts to isolate the virus from the faeces and tissues from Pteropus spp. were unsuccessful. Conclusion Serological evidence of infection with MenV was detected in Pteropus spp. in Australia. Although virus‐like particles were detected in faeces, no viruses were isolated from faeces, urine or tissues of Pteropus spp.     

Non-specific seroreactions against Brucella abortus in ruminants in New Zealand and the presence of Yersinia enterocolitica 0:9

The level of non-specific reactions found in the brucellosis serology of ruminants in New Zealand was very low until July 1992. This changed when, in an export consignment of 1071 deer, 35% reacted in the Brucella abortus tube agglutination test with titres varying from 50 to 200 IU. The reactors were also positive in the Rose-Bengal agglutination test and most of them reacted in the complement fixation test with titres varying from 10 to 80 IU. Yersinia enterocolitica 0:9 was later isolated from one deer of this consignment. It was the first isolate of this serotype recovered in New Zealand from an animal. Shortly after, false reactors occurred more frequently than before in sera from Brucella abortus accredited free cattle herds. As the involvement of Yersinia enterocolitica 0:9 was suspected in these cases, faecal samples from reactors and in-contact animals were cultured for this organism. Yersinia enterocolitica 0:9 was isolated from nine of 19 herds showing one or more false Brucella abortus seroreactions.

Prior to 1990, Yersinia enterocolitica serotype 0:9 had not been isolated in New Zealand, despite the recovery of a number of other bio— or serotypes of the organism from humans and animals. From 1990 onward, serotype 0:9 began to be isolated from human faecal samples with increasing frequency. Since the first isolations from deer and cattle in 1992, it has now also been recovered from a cat and an alpaca and from cattle without any association with false positive Brucella abortus reactions. All serotype 0:9 isolates were of biotype 2.