Embryo transfer from cattle infected with bluetongue virus

Embryos recovered nonsurgically from donor cattle during the peak of bluetongue viremia were surgically transferred to seronegative recipients 7 to 8 or 10 to 11 days after the onset of donor estrus. Virus was isolated from the uterine flushing medium recovered from 11 of the 20 donors. Bluetongue virus was not isolated from the blood of any of 39 recipients, nor did any recipient seroconvert to the virus following transfer. The number of recipients that became pregnant after transfer of embryos from infected donors (21 of 39) was not significantly different from contemporary controls. Virus antigen was not detected by immunofluorescence in any of 63 embryos and oocytes recovered from viremic donors. These results indicate that under standard embryo transfer conditions, transmission of bluetongue virus from infected donors to uninfected recipients is unlikely to occur.     

Regional seroprevalence of bluetongue virus in cattle in Illinois and western Indiana

OBJECTIVE: To estimate seroprevalence of bluetongue virus (BTV) and the geographic distribution of seropositive cattle herds in Illinois and western Indiana. SAMPLE POPULATION: 10,585 serum samples obtained from cattle in 60 herds during 3 transmission seasons (2000 through 2002). PROCEDURES: In a longitudinal study, serum samples were tested for BTV antibodies by use of a competitive ELISA. Four geographic zones were created by use of mean minimum January temperature. A multivariable mixed-effects logistic regression model with a random effect for herd was used to estimate seropositive risk for zone, age of cattle, herd type, and transmission season. RESULTS: Overall, BTV antibodies were detected in 156 (1.5%) samples. Estimated seroprevalence in 2000, 2001, and 2002 was 1.49%, 0.97%, and 2.18%, respectively. Risk of being seropositive for BTV was associated with geographic zone and age. Seroprevalence increased progressively from northern to southern zones, with no evidence of BTV infection in the northernmost zone. In the southernmost zone, annual seroprevalence ranged from 8.65% to 11.00%. Adult cattle were 2.35 times as likely as juvenile cattle to be seropositive. CONCLUSIONS AND CLINICAL RELEVANCE: Overall seroprevalence was lower than has been reported for Illinois cattle. Bluetongue virus antibodies were distributed heterogeneously in this region. Only in the southernmost zone was seroprevalence consistently > 2%. Regionalization of BTV risk based on state borders does not account for such variability. Serologic data could be combined with landscape, climate, and vector data to develop predictive models of BTV risk within transitional regions of the United States.     

Serogrouping of United States and some African serotypes of bluetongue virus using RT-PCR

The diagnostic potential of RT-PCR for detection of bluetongue virus (BTV) ribonucleic acid (RNA) sequence in cell culture and tissue samples from infected ruminants from United States, Sudan, South Africa and Senegal, was evaluated. The non structural protein 1 (NS1) gene of North American BTV serotype 11 was targeted for PCR amplification. The United States BTV serotypes 2, 10, 11, 13 and 17 and the Sudanese BTV serotypes 1, 2, 4 and 16 and BTV serotype 4 from South Africa and BTV serotype 2 from Senegal were studied. RNAs from all BTV field isolates used in this study, propagated in cell cultures, were detected by the described RT-PCR-based assay. The first specific 790bp BTV PCR products were amplified using a pair of outer primers (BTV1 and BTV2). Specificity of the PCR products was confirmed by a nested amplification of a 520bp PCR product using a pair of internal (nested) primers (BTV3 and BTV4). The BTV PCR products were visualized on ethidium bromide-stained agarose gels. Amplification products were not detected when the RT-PCR-based assay was applied to RNAs from closely related orbiviruses including, epizootic hemorrhagic disease virus (EHDV) prototypes serotypes 1, 2, 4; RNA from Sudanese isolate of palyam orbiviruses serogroup and total nucleic acid extracts from uninfected Vero cells. Application of the nested BTV RT-PCR to clinical samples resulted in amplification of BTV RNA from blood and serum samples from goats experimentally infected with BTV4 and from naturally infected sheep, goats, cattle and deer. The results of this study indicated that this RT-PCR assay could be applied for rapid detection of BTV, in cell culture and clinical samples from susceptible ruminants during an outbreak of the disease, in the United States and African.     

Interaction of bluetongue virus with preimplantation embryos from mice and cattle

Preimplantation embryos from mice and cattle were exposed to bluetongue virus in vitro to determine whether the virus would replicate in these early embryos and, if so, what pathologic consequences would ensue. A high proportion of zona pellucida-free, 2-cell embryos and morulae from mice, and morulae from cattle became infected. The infection was rapidly cytopathic in embryos from both species. Indirect immunofluorescence was used to demonstrate accumulation of virus antigen in the blastomeres of these embryos. The zona pellucida of both murine and bovine embryos provided effective protection from virus present in culture fluid.     

Recovery of dual serotypes of bluetongue virus from infected sheep and cattle

Dual serotypes of bluetongue virus (BTV) were recovered from field-collected samples of sheep and cattle blood. Two sheep, each infected with both BTV serotypes 10 and 17, were found in a flock with bluetongue disease associated with these two serotypes. One sheep infected with BTV serotypes 11 and 17 was found in a second flock; it was the only viremic sheep detected and was clinically ill. Dual serotype infections of one beef and two dairy cattle were found in three geographically separate herds; mixtures recovered were of BTV serotypes 10 and 17 and serotypes 11 and 17. Clinical signs of illness were absent in the cattle in two herds, but severe conjuctivitis was seen in several cows in a third herd, including the cow with a dual serotype infection (BTV 11 and 17). Two of the cattle with dual infections had no serological evidence of BTV as determined by the agar gel precipitin test; serum was not available from the other cow with a dual serotype infection. The significance of dual infections and immune tolerance are discussed.     

Recombinant cDNA probe from bluetongue virus genome segment 10 for identification of bluetongue virus

Genome segment 10 of bluetongue virus (BTV) serotype 11 UC8 strain was cloned and subsequently hybridized to viral double-stranded RNA extracted from 90 field isolates of BTV serotypes 10, 11, 13, and 17; the prototype strains of BTV 2, 10, 11, 13, and 17; the prototype strain epizootic hemorrhagic disease virus (EHDV) serotype 1; and 4 field isolates of EHDV serotype 2. The 90 field isolates were obtained from different counties in California, Louisiana, and Idaho during the years 1979, 1980, and 1981. The cloned genetic probe hybridized with all the BTV samples tested, showing different degrees of cross-hybridization at the stringency conditions used in this study. This indicated that BTV genome segment 10 has conserved nucleotide sequences among the BTV serotypes 2, 10, 11, 13, and 17. No cross-hybridization signals were detected between the cloned genome segment 10 of BTV 11 UC8 strain and the prototype strain of EHDV serotype 1 and the field isolates of serotype 2. This probe recognized a wide variety of BTV isolates.     

进境蓝舌病抗体阳性动物带毒分析及检疫处理研究

为评估进口蓝舌病病毒(BTV)抗体阳性动物的感染状态,分析其带毒风险,对从国外进口的9批19714头动物,无菌采集全血分离血清,采用竞争ELISA方法检测BTV抗体。对BTV抗体检测阳性的动物,采集抗凝血,采用OIE推荐的套式RT-PCR和荧光RT-PCR方法进行检测,同时将样品送往蓝舌病参考实验室进行病毒分离鉴定,以确定动物的蓝舌病感染状态。结果 9批动物中检出28头BTV抗体阳性动物,但核酸检测和病毒分离鉴定的结果均表明这些BTV抗体阳性动物并不携带有非感染性和感染性病毒粒子。结合9个批次的进口动物并无明显临床表现,且进口动物来源地也无蓝舌病(BT)的疫情发生,据此根据OIE《陆生动物卫生法典》条款,判定这些抗体阳性动物为带毒阴性。本研究通过对BTV抗体阳性动物的带毒分析,并结合OIE确定BTV感染的要求,对BTV口岸隔离检疫流程提出建议,以指导动物检疫工作,阻止病原经口岸传入。     

Isolation of an exotic serotype of bluetongue virus from imported cattle in quarantine.

In 1980, 60 zebu cattle from Brazil were admitted into quarantine in Florida for 150 days. During the 30 days between their last test in Brazil and their first test in Florida, four animals developed antibody to bluetongue virus detectable by agar gel immunodiffusion test. Within 62 days after arrival in Florida, three more seroconverted and one more was positive by the 86th day. Virus neutralizing titers of serums from the first four cattle were highest against bluetongue virus serotype 4 and 20; both of these serotypes are exotic to the United States. A bluetongue virus serotype 4 was isolated from one of these animals. The eight positive reactors were slaughtered; the other 52 cattle, which did not develop detectable antibody titers to bluetongue virus, were released into the United States.     

Temporal appearance, geographic distribution, and species of origin of bluetongue virus serotypes in the United States.

Beginning in 1973, all available laboratory and field strains of bluetongue virus (BTV) from the United States were serotyped. Of the viral strains serotyped, 27 were collected from 1953 through 1972; 173 were collected from 1973 through 1977. Although 20 BTV serotypes have been found worldwide, only BTV serotypes 10, 11, 13, and 17 have been found in the United States. Since 1973, serotypes 11 and 17 have been the prevalent serotypes. Samples were collected over a 24-year period in the United States and represent a wide geographic area and diverse host sources (sheep, cattle, wild ruminants, and insect vectors). The collection was not a statistical sampling.     

Susceptibility of Sudanese sheep to a bluetongue virus isolated from apparently healthy cattle in the Sudan

This study intends to clarify the role of apparently healthy cattle as a reservoir of bluetongue (BT) virus to sheep in the Sudan. It confirms earlier work and establishes that cattle can harbour bluetongue virus to which sheep are susceptible in the country. Experimental transmission of BT virus between the two species suggests that the best indicator to determine viraemia in apparently healthy cattle is to inoculate susceptible sheep with suspected cattle virus. The condition of the viraemia and the virus survival in the field are discussed.     

Nontransmission of bluetongue virus by embryos from bluetongue virus-infected sheep

Donor sheep were infected either by bites of bluetongue virus (BTV)-infected (serotype 11, "Texas Station strain") Culicoides variipennis or by inoculation with 100,000 median chicken embryo intravascular lethal doses of BTV (serotype 11) from a suspension made from infected C variipennis. Fourteen embryos from 4 BTV-infected ewes bred by rams not infected with BTV were transferred to 8 BTV-seronegative recipient ewes, and 35 embryos and 4 unfertilized eggs from 14 BTV-infected ewes bred by BTV-infected rams were transferred to 19 BTV-seronegative recipient ewes. Eleven pregnancies and 12 lambs resulted. None of the recipients or lambs seroconverted, and BTV was not isolated from the pregnant recipient ewes or their lambs at slaughter 30 days after parturition.