Detection of bovine viral diarrhea virus (BVDV) in seropositive cattle

Detection of bovine virus diarrhoea virus (BVDV) in one vaccinated beef cattle and three non-vaccinated dairy herds was investigated on peripheral blood leukocytes (PBL) with or without previous treatment followed by a capture ELISA (cELISA). Using the combination of PHA and polycation treatment, PBL from 229 seropositive cattle were studied and could be classified in four different states of BVDV infection. Lysed PBL from four animals were directly positive in cELISA (Category I), PBL of 17 animals were positive after PHA stimulation (Category II), 15 animals were positive only after PHA stimulation plus polycation treatment (Category III), while virus could not be detected in 193 seropositive cattle. Wild-type BVDV strains were isolated by co-culture on polycation-treated MDBK cells from 11 of these seropositive animals. BVDV antibodies of these same animals were able to neutralize their own virus, indicating that virus persists in PBL in spite of strain-specific antibodies. No apparent change of leukocyte subpopulations could be detected in any category of virus-positive animals. Thus, BVDV may be present in the PBL of some cattle, even in the presence of a specific active immune response.     

Natural changes in the spread of bovine viral diarrhoea virus (BVDV) among Estonian cattle

The results of a survey conducted during 1993-2000 to study the spread of bovine viral diarrhoeal virus (BVDV) among Estonian cattle are presented. The BVDV infection status of a representative random sample of cattle herds housing 20 or more dairy cows was established to estimate the prevalence of herds with active BVDV infection [potentially having persistently infected (PI) cattle--suspect PI herds]. The herds investigated comprised approximately 70% of all Estonian dairy cows. The BVDV infection status was established in 315-350 herds (making the sampling fraction about 20%) during three sampling periods: 1993-95, 1997-98, 1999-2000. BVDV antibodies were detected in herd bulk milk samples and/or sera from young stock by a liquid-phase-blocking enzyme-linked immunosorbent assay developed in the Danish Veterinary Institute for Virus Research. The results of the survey demonstrate the reduction in the prevalence of herds with active BVDV infection in the studied fraction of the Estonian cattle population. During the first sampling period (1993-95) a prevalence of 46% (+/- 5%) for suspect PI herds was observed, during the second sampling period this prevalence was 16% (+/- 3%) and in the third period it was 18% (+/- 3%). As there is no control programme for BVDV in Estonia, the observed changes reflect the natural course of the infection in the study population. A possible cause for these changes is the decreased trade in breeding animals as a result of the economic difficulties present in cattle farming during the study period. The farming practices (most large herds are managed as closed herds) and the low density of cattle farms have obviously facilitated the self-clearance of herds from the BVDV infection, diminishing the new introduction of infection into the herds.     

Preliminary studies on enzootic bovine leukosis in Saudi dairy farms

The occurrence of sporadic cases of enzootic bovine leukosis in commercial dairy farms in Saudi Arabia was recently confirmed and found to be associated with importation of breeding heifers. Immunodiffusion test was applied to screen the prevalence of infection with bovine leukemia virus among local traditional and dairy cattle. All the 102 examined local cattle were negative, while out of the 1329 tested dairy animals (originating from 23 farms), 268 (from 16 farms) showed precipitating activity. As an epizootiological model, all animals of an infected dairy farm were serologically examined. Out of the 560 originally imported cows and the 1849 animals born locally in the farm, 217 (39%) and 468 (25%) animals, respectively, were found positive. The correlation between the age of locally born animals and the occurrence of antibodies against bovine leukemia virus was discussed.     

Prevalence of BVDV infection in Greek dairy herds

Thirty-nine Greek dairy herds, totalling 6333 cattle, enrolled in a voluntary bovine viral diarrhoea virus (BVDV) eradication programme based on the identification and removal of persistently infected (PI) animals. The aim of this study was to estimate the prevalences of BVD antigen-positive and PI animals, and investigate the significance of the associations between the prevalence estimates and herd size. Initially, all animals were bled and examined for BVDV, using an antigen ELISA. A second sample was collected from the positive animals, after a period of at least three weeks. Animals retested positive were classified as PI. Antigen positive and PI animals were detected in all herds. The respective mean prevalences, adjusted for the test's accuracy and the herd-clustering effect, were 14% (95%CI: 11-18%) and 1.3% (0.8-1.8%), respectively. Herd size was not associated with the prevalence of antigen-positive or PI animals.     

Prevalence of antibody to rotavirus in Moroccan cattle

A serological survey was carried out in order to determine the prevalence of antirotavirus antibodies in Moroccan cattle under different management conditions. From the 493 serum samples examined, 325 (65.9%) were found positive, using a counter-immunoelectroosmophoresis technique. Animals of indigenous breed coming from farms with rapid turnover or large number of animals, or having frequent contacts with imported cattle, had a higher rate of seropositivity; however, positive sera were also found in cattle from small farms in remote areas, showing that rotavirus infection is ubiquitous in that country. No relationship was found between the prevalence of anti-rotavirus antibodies and the frequence of calf diarrhoea. The percentage of seropositive animals in a herd has to be considered as an epidemiological indicator.     

Prevalence and epidemiological features of bovine viral diarrhoea virus infection in Lithuania

The objectives of the present work were to estimate the level of bovine viral diarrhoea virus (BVDV) infection in cattle herds at the different Lithuanian districts and to determine factors influencing the course of BVDV infection. The studies were explored in 147 intensive dairy cattle breeding herds from 27 different Lithuanian regions in 1997-2001. BVDV infection was diagnosed in all investigated regions. The existing variations in the structure of cattle population determined different distribution patterns of BVDV infection. The number of seropositive animals ranged from 11.9 to 100%. It must be pointed out that 29.9% of the herds were not infected with BVDV and in 32.7% of the herds from 70 to 100% of cattle were seropositive to BVDV. A positive correlation between the number of seropositive cattle, and the size of herds and age of animals was determined. Sex of animal had no influence on the prevalence of BVDV. It was estimated that the annual incidence risk of infection with BVDV decreases with the animal age.     

Field application of the combination of neutralizing test and virus isolation, so-called spot test, to detect herds including cattle persistently infected with bovine viral diarrhea virus

To detect herds including cattle persistently infected (PI) with bovine viral diarrhea virus (BVDV), application of the combination of neutralizing antibody detection and virus isolation, so-called spot test, were performed on sera of 3 calves selected from each of 26 farms. Nine farms were judged as positive because 64 or more antibody titers were detected from 2 or more calves or BVDV was isolated from one or more calves. PI cattle were detected from 8 of the 9 farms. The positive judgment on one farm was obtained only when the indicator virus used on the neutralizing test was genotypically identical with the isolate from the farm. These results suggest that the spot test can be effective in detecting herds with PI cattle and that the accuracy may be influenced by the genotypes of the indicator viruses.     

Evaluation of long-term antibody responses to two inactivated bovine viral diarrhoea virus (BVDV) vaccines

The aim of the present study was to determine the serological response of heifers after vaccination with two inactivated bovine viral diarrhoea virus (BVDV) vaccines by means of various ELISA tests. Three dairy farms were selected from the Galicia region of Spain. In each herd, a batch of heifers to be vaccinated for the first time was selected and followed for 15 months. Heifers from farm 1 (n = 25) were vaccinated with Vaccine A, whereas heifers from farm 2 (n = 16) were vaccinated with Vaccine B. Heifers from farm 3 (n = 17), where no BVDV vaccines were used, acted as controls. Blood samples were analyzed periodically for BVDV antibodies, using five commercial ELISAs, based on BVDV p80 antigen or whole virus.At the end of the study, none of the animals vaccinated with Vaccine A seroconverted according to p80 antibody status, whereas up to 80% tested positive by ELISA against whole virus antigen. For the animals vaccinated with Vaccine B, 2/16 animals seroconverted according to p80 antibody ELISAs, whereas all had seroconverted according to the ELISA against whole virus antigen. In most cases, based on the use of ELISAs to detect specific antibodies against the p80 protein, at 15 months post-vaccination with inactivated BVDV vaccines the responses did not seem to interfere with detection of antibody to BVDV infection. However, the finding of a small proportion of vaccinated animals seropositive against BVDV p80 antigen suggests that antibodies that interfere with diagnosis of BVDV infection within the herd could exist, even when using p80 ELISAs.     

Seroconversion to bovine viral diarrhoea virus and infectious bovine rhinotracheitis virus in dairy herds of Michoacan, Mexico

Bovine viral diarrhoea virus (BVDV) and infectious bovine rhinotracheitis virus (IBRV) are important viral diseases around the world. The objective of this study was to estimate the incidence of seroconversion to BVDV and IBRV and to identify associated risk factors in dairy herds of Michoacan, Mexico. The longitudinal study included 62 herds and ran from December 2001 to November 2002. The total number of animals enrolled and completing the study were 392 and 342 animals for BVDV and 925 and 899 animals for IBRV. Animals were tested monthly for 12 months, for the presence of antibodies. Risk factors were: herd size (2–9, 10–25 and 26–55 animals), herd serostatus (seropositive or seronegative, only for IBRV), age group of the animal (6 to 12, 13 to 24, 25 to 48 and > 48 months) and animal origin (born in farm, purchased). The cumulative incidences for BVDV and IBRV were 16.4% and 3.4%, respectively; whereas, the incidence density rates for BVDV and IBRV were 15.9 and 2.9 per 1000 animal-months at risk, respectively. Seroconversion curves were statistically different for age group for BVDV and IBRV and for herd status for IBR. The relatively high incidence of seroconversion for BVDV suggests that a successful control programme should be oriented towards the identification and elimination of the PI animals and towards avoiding the introduction of PI cattle to the farm. The scenario of IBRV is favourable to implement a programme directed to reduce the number of new seropositive herds.     

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.     

新疆部分规模奶牛场牛病毒性腹泻病的流行情况调查与防控措施

为了解新疆地区部分规模奶牛场牛病毒性腹泻病（BVD）的流行情况,优化防控措施,以达到建设防控净化场的目的,自2020年11月到2021年7月累计采集新疆5 个地区16 个奶牛场共计26 997 份血清、3 843 份犊牛耳组织进行全群普检。通过使用IDEXX公司牛病毒性腹泻病毒（BVDV）抗原检测试剂盒检测及RT-PCR复检结合测序等方法,淘汰阳性牛,同源性分析流行毒株情况。BVDV血清抗原检测结果为：沙湾某奶牛场BVD阳性率为1.63%（38/2 326）;乌鲁木齐某奶牛场BVD阳性率为0.35%（4/1 132）;其余奶牛场均为阴性。犊牛耳组织抗原检测结果为：乌鲁木齐某牛场BVDV阳性率为1.17%（4/342）;其余奶牛场均为阴性。研究结果揭示,奶牛场通过淘汰BVDV阳性牛,调整免疫程序、制定消毒程序等方法,可有效净化BVD。     

Multiple diagnostic tests to identify cattle with Bovine viral diarrhea virus and duration of positive test results in persistently infected cattle

Several tests for Bovine viral diarrhea virus (BVDV) were applied to samples collected monthly from December 20, 2005, through November 27, 2006 (day 0 to day 342) from 12 persistently infected (PI) cattle with BVDV subtypes found in US cattle: BVDV-1a, BVDV-1b, and BVDV-2a. The samples included clotted blood for serum, nasal swabs, and fresh and formalin-fixed ear notches. The tests were as follows: titration of infectious virus in serum and nasal swabs; antigen-capture (AC) enzyme-linked immunosorbent assay (ELISA), or ACE, on serum, nasal swabs, and fresh ear notches; gel-based polymerase chain reaction (PCR) testing of serum, nasal swabs, and fresh ear notches; immunohistochemical (IHC) testing of formalin-fixed ear notches; and serologic testing for BVDV antibodies in serum. Of the 12 animals starting the study, 3 died with mucosal disease. The ACE and IHC tests on ear notches had positive results throughout the study, as did the ACE and PCR tests on serum. There was detectable virus in nasal swabs from all the cattle throughout the study except for a few samples that were toxic to cell cultures. The serum had a virus titer ≥ log₁₀ 1.60 in all samples from all the cattle except for 3 collections from 1 animal. Although there were several equivocal results, the PCR test most often had positive results. The BVDV antibodies were due to vaccination or exposure to heterologous strains and did not appear to interfere with any BVDV test. These findings illustrate that PI cattle may be identified by several tests, but differentiation of PI cattle from cattle with acute BVDV infection requires additional testing, especially of blood samples and nasal swabs positive on initial testing. Also, calves PI with BVDV are continual shedders of infectious virus, as shown by the infectivity of nasal swabs over the 11-mo study.     

Prevalence and geographic distribution of bovine viral diarrhoea (BVD) infection in Finland 1993-1997

Bulk milk samples from every herd supplying milk to dairies in Finland were examined for the presence of antibodies to BVD virus (BVDV) annually during 1993-1997. The highest prevalence, 0.99% in 1994, declined to 0.37% in 1996; however, this favourable trend appeared to discontinue in 1997, where the prevalence remained at 0.41%. In 1993, sera of all individual animals from bulk milk antibody-positive herds were examined for the presence of these antibodies. Since 1994, only sera of animals from herds with a bulk milk absorbance reading greater than 0.250 in the EIA test were examined individually. Three geographic foci of BVDV antibody-positive dairy herds were resolved in 1994, one in the north-western, another in the eastern and a diffuse third in the southern part of Finland. A distinct limiting of the spread was apparent in 1997. Beef cattle were also studied during 1993-1997; in 1993 breeding units, in 1994 mainly beef suckler herds and in 1995-1997 serum samples of beef animals at slaughter were examined for the presence of antibodies to BVDV. The prevalence of seropositive herds in 1993 and 1994 was 30.2% and 3.2%, respectively, while the prevalence among slaughter animals ranged 0.8-1.6%. Seronegative animals in herds with > 50% of seropositive animals were examined for the presence of BVD-virus. A total of 40 dairy herds and two beef herds with viraemic (persistently infected, PI) animals was encountered during 1993-1997. A comprehensive control programme and a more specific, cooperatively funded eradication programme for dairy cattle were launched in 1994. These programmes most probably contributed to the decline in prevalence during 1994-1996.     

Description of risk factors associated with the detection of BVDV antibodies in Brazilian pig herds

Bovine viral diarrhea virus (BVDV) infects ruminants as primary hosts. However, other animals like pigs are susceptible. This study was conducted to investigate seroprevalence and risk factors associated with the detection of BVDV antibodies in pig herds. A total of 1.705 serum samples of 33 finisher herds, from seven Brazilian states, were collected in slaughterhouses. The samples were tested by virus neutralization (VN) test. In total, 5.35% (91/1.705) were positive and 64% (21/33) of the herds had positive animals. A significant association with “trucks are not cleaned and disinfected” and “visitors do not respect 72-h interval between visits to farms” (P?<?0.05) was found in association with detection of BVDV-2 antibodies. This study suggests that important biosecurity gaps are present in Brazilian pig farms, as the presence of BVDV antibodies in pigs suggests (direct or indirect) contact with population(s) of ruminant species. Closing biosecurity gaps prevents spread of BVDV and other pathogens such as foot-and-mouth disease virus (FMDV) between pig and ruminant farms. This data should be taken in account by CSF surveillance programs, once cross-reaction in serologic tests between classical swine fever virus (CSFV) and BVDV antibodies has been shown to occur.     

Studies on the Decline of Bovine Virus Diarrhoea Virus (BVD V) Maternal Antibodies and Detectability of BVDV in Persistently Infected Calves

Persistently infected (PI) animals play a significant role in spread and transmission of bovine virus diarrhoea virus (B VD V) (Duffell & Harkness 1985). The identification and removal of PI cattle from the herd is of great importance in the control of BVDV. Although PI animals often show various degrees of growth retardation and unthrifty appearance, a significant proportion is clinically normal. PI animals are often seronegative (Duffell & Harkness 1985), but calves may be tested seropositive because of the presence of maternal immunity (Meyling & Jensen 1988). The passively derived BVDV antibodies may interfere with the ability to detect virus. Considering the importance of early recognition of PI calves, it is essential to determine the earliest time when PI animals can safely be diagnosed in the herd.     

Seroprevalence of OHV-2, BVDV, BHV-1, and BRSV in ranch-raised bison (Bison bison).

Serum samples were collected at slaughter from 226 24-30-month-old American bison (Bison bison) bulls from Kansas, Minnesota, North Dakota, and Manitoba and assayed for antibodies to ovine herpesvirus type-2 (OHV-2), bovine viral diarrhea virus (BVDV), bovine herpesvirus type-1 (BHV-1), and bovine respiratory syncytial virus (BRSV). Antibodies were detected by serum neutralization for BVDV, BHV-1, and BRSV, while antibodies to OHV-2 were detected by competitive inhibition-ELISA (CI-ELISA). Detectable antibodies were found against all viruses: 10 of 226 (4.40%) against OHV-2, 125 of 226 (55.3%) against BVDV, 99 of 226 (43.8%) against BHV-1, and 208 of 226 (92.0%) against BRSV. Titers from 93.6% of the BVDV-positive animals, 79.8% of the BHV-1-positive animals, and 98.1% of the BRSV-positive animals were > or = 1.25. These data indicate that a low percentage of clinically normal bison are seropositive for OHV-2 while a high percentage of bison sampled are seropositive for BVDV, BHV-1, and BRSV.     

Response of cattle persistently infected with bovine virus diarrhoea virus to bovine leukosis virus

Six cattle persistently infected with bovine virus diarrhoea virus (BVDV) and seronegative, and two control, virus negative seropositive cattle were inoculated with lymphocytes infected with bovine leukosis virus (BLV). The two controls produced a normal immune response to BLV, developing antibodies at four and five weeks after inoculation. Two of the six cattle persistently infected with BVDV developed a strong antibody response by six weeks after inoculation with BLV. Four developed a depressed response to BLV, characterised in three by a 'hooking' reaction in the immunodiffusion test which persisted in successive bleedings but was interspersed occasionally by a weak positive reaction. In one of these animals, a series of 'hooking' reactions was followed by a number of negative results. The fourth animal remained serologically negative until 16 weeks after inoculation when a 'hooking' reaction was observed followed by a series of negative results. BLV was isolated from all the cattle persistently infected with BVDV at 42 or 58 weeks after inoculation regardless of whether the serum samples gave negative, 'hooking', weak positive or positive reactions in the immunodiffusion test. BLV was consistently isolated from the nasal secretions of a steer which was BVDV negative but seropositive. The possibility of decreased immune responsiveness to BLV in animals persistently infected with BVDV should be considered when formulating regulations governing the testing of animals for freedom from BLV.     

Herd-level risk factors for Neospora caninum seroprevalence in dairy farms in southern Brazil

A cross-sectional study was used to test the relationship between herd seroprevalence to Neospora caninum and various potential herd-level risk factors in 60 dairy farms located in two distinct regions in southern Brazil. Thirty farms were randomly selected from within each region. A questionnaire was designed to summarize each farm's production system as it might relate to N. caninum transmission. The questionnaire contained 105 closed questions relating to general characteristics of the farms, farm facilities, management, source of food and water, herd health, environment and biosecurity, which included questions relevant to N. caninum transmission, including presence and number of dogs and other animals, purchase of animals and contact with man. Serum samples were collected from 40% of animals in each farm and N. caninum antibodies were detected by immunofluorescent antibody test (IFAT). The association between potential risk factors and the probability of an animal being seropositive was modeled using a generalized estimation equations (GEE) logistic regression model. The model accounted for multilevel correlation of data from multiple animals within herds. The mean (+/-S.D.) number of animals in the 60 herds was 64.5 (+/-45.6), ranging from 20 to 280 females. Blood samples were collected from 1549 animals. The size of the farms varied from 4 to 100 ha (mean 30.1+/-25.9 ha). At least one dog was found in 57 of the 60 dairy farms (95%). The mean number of dogs was 3.1 (+/-1.9), ranging from 0 to 10. All females were raised on pasture. For all cattle sampled, N. caninum seroprevalence was 17.8%. Overall, 93.3% of herds (56/60) had at least one seropositive animal identified. Four variables were significantly associated with N. caninum sero-response in the 57 dairy farms, which were included in the final multivariable model: the number of dogs on the farm, farm area (hectares), feeding pooled sources of colostrum and region. The odds of a cow being seropositive increased 1.13 times for each additional dog present on the farm (P=0.021). Cattle from farms that fed calves colostrum pooled from multiple cows had 1.79 times greater odds for being seropositive for N. caninum (P<0.003). The probability of being seropositive was inverse to the area of the farms, such that cattle had 0.92 times the odds to be seropositive (P=0.014) for each additional 10 ha of farmland. Finally, cattle from farms in region one had 0.71 times the odds to be seropositive than cattle from region two (P=0.035). Results of this study suggest that several risk factors may explain why dairy cattle in Brazil may become exposed to N. caninum. However, further investigation of these factors is necessary because the purpose of this study was to refine and generate hypotheses on N. caninum transmission.     

Bovine virus diarrhoea (BVD) control programme in an area in the Rome province (Italy)

A BVD control programme based on the identification and removal of persistently infected (PI) animals is being undertaken in an area in the Rome province, where BVD outbreaks had been previously detected. It involves 174 mainly dairy herds, from which blood samples of all bovines older than 1 year are obtained through the national brucellosis and leukosis eradication programme. Samples sufficient to detect the presence of seropositive animals at a prevalence of 5% or more are initially screened for antibodies against BVD virus (BVDV) using an immunoenzymatic assay. Upon identification of seroreagents additional blood samples are tested from the 6-12-month age category not included in the initial samples. Animals are considered immunotolerant if BVDV is demonstrated twice at a minimum 30-day interval. When no seropositive animals are detected during the first serological screening the herd is declared BVD-free if a second testing, preferably carried on the same animals previously tested, confirms the seronegative status of the herd. At present 147 farms have been tested, of which 63 (42.9%) are negative with respect to antibodies against BVDV. Of the 84 remaining herds in which one or more seropositives are detected, 13 are classified as recently infected. In eight of these recently infected herds, 22 PI animals have been identified.