The increasing threat of vector-borne diseases (VBDs) represents a great challenge to those who manage public and animal health. Determining the spatial distribution of arthropod vector species is an essential step in studying the risk of transmission of a vector-borne pathogen (VBP) and in estimating risk levels of VBD. Risk maps allow better targeting surveillance and help in designing control measures. We aimed to study the geographical distribution of Culicoides imicola, the main competent vector of Bluetongue virus (BTV) in sheep in Tunisia. Fifty-three records covering the whole distribution range of C.imicola in Tunisia were obtained during a 2-year field entomological survey (August 2017 - January 2018 and August 2018 - January 2019). The ecological niche of C. imicola is described using ecological-niche factor analysis (ENFA) and Mahalanobis distances factor analysis (MADIFA). An environmental suitability map (ESM) was developed by MaxEnt software to map the optimal habitat under the current climate background. The MaxEnt model was highly accurate with a statistically significant area under curve (AUC) value of 0.941. The location of the potential distribution of C. imicola is predicted in specified regions of Tunisia. Our findings can be applied in various ways such as surveillance and control program of BTV in Tunisia.

The increasing threat of vector-borne diseases (VBDs) represents a great challenge to those who manage public and animal health. Determining the spatial distribution of arthropod vector species is an essential step in studying the risk of transmission of a vector-borne pathogen (VBP) and in estimating risk levels of VBD. Risk maps allow better targeting surveillance and help in designing control measures. We aimed to study the geographical distribution of Culicoides imicola, the main competent vector of Bluetongue virus (BTV) in sheep in Tunisia. Fifty-three records covering the whole distribution range of C.imicola in Tunisia were obtained during a 2-year field entomological survey (August 2017 – January 2018 and August 2018 – January 2019). The ecological niche of C. imicola is described using ecological-niche factor analysis (ENFA) and Mahalanobis distances factor analysis (MADIFA). An environmental suitability map (ESM) was developed by MaxEnt software to map the optimal habitat under the current climate background. The MaxEnt model was highly accurate with a statistically significant area under curve (AUC) value of 0.941. The location of the potential distribution of C. imicola is predicted in specified regions of Tunisia. Our findings can be applied in various ways such as surveillance and control program of BTV in Tunisia.

Replication of bluetongue virus (BTV) in leukocytes from the blood of sheep, cattle, elk, and mule deer inoculated with BTV serotype 10 or 17 was assessed by immunocytochemical staining and dot blot northern hybridization to determine if differences in the prevalence of infection in this blood fraction might account for the differences in clinical disease among these species. Viremia was confirmed by virus isolation in all inoculated animals. Analysis of leukocytes with monoclonal antibodies specific for BTV proteins revealed low numbers of infected leukocytes in only 2 sheep 8 days after inoculation with BTV serotype 10. Most of the cells expressing BTV were identified morphologically as monocytes; approximately 10% of infected cells were lymphocytes. Bluetongue virus was not detected by use of dot-blot hybridization on samples of blood. Our results suggest that differential infection of leukocytes does not account for the pronounced differences in clinical signs and pathologic changes among ruminants.

Three independent 1-year studies were conducted during 3 consecutive years to better define the prevalence of bluetongue virus (BTV) infection in Mexico. Serologic data were obtained by use of agar-gel immunodiffusion for identification of BTV group-reactive antibodies, and virologic data were obtained by virus isolation. Samples were obtained from sheep in 6 states over a 1-year period, with 9% seropositive; samples were obtained from cattle in 11 states during the same 1-year period, with 35% seropositive. Two years later, samples were obtained from cattle in 4 additional states, with 69% seropositive. Virus isolation was conducted on pooled blood samples obtained from cattle in 7 states. Six virus isolates were recovered and included 2 isolates each of BTV serotypes 11 and 13 and 1 isolated each of serotypes 10 and 17. All virus isolates were partially characterized by electrophoretic analysis of genomic RNA migration profiles (electropherotypes) in polyacrylamide gels. All Mexican isolates of BTV differed considerably in electropherotype profile, as compared with their respective US prototype strain of the same serotype. Such differences appeared to be much more extensive than those described to exist between numerous California isolates of the same serotype.

Bluetongue (BT), a disease that affects mainly sheep, causes economic losses owing to not only its deleterious effects on animals but also its associated impact on the restriction of movement of livestock and livestock germplasm. The causative agent, bluetongue virus (BTV), can occur in the semen of rams and bulls at the time of peak viraemia and be transferred to a developing foetus. The risk of the transmission of BTV by bovine embryos is negligible if the embryos are washed according to the International Embryo Transfer Society (IETS) protocol. Two experiments were undertaken to determine whether this holds for ovine embryos that had been exposed to BTV. Firstly, the oestrus cycles of 12 ewes were synchronised and the 59 embryos that were obtained were exposed in vitro to BTV-2 and BTV-4 at a dilution of 1 x 102.88 and 1 x 103.5 respectively. In the second experiment, embryos were recovered from sheep at the peak of viraemia. A total of 96 embryos were collected from BTV-infected sheep 21 days after infection. In both experiments half the embryos were washed and treated with trypsin according to the IETS protocol while the remaining embryos were neither washed nor treated. All were tested for the presence of BTV using cell culture techniques. The virus was detected after three passages in BHK-21 cells only in one wash bath in the first experiment and two unwashed embryos exposed to BTV-4 at a titre of 1 x 103.5. No embryos or uterine flush fluids obtained from viraemic donors used in the second experiment were positive for BTV after the standard washing procedure had been followed. The washing procedure of the IETS protocol can thus clear sheep embryos infected with BTV either in vitro or in vivo.

To evaluate herd-level risk factors for seropositive status of cattle to 1 or more bluetongue viruses. 110 herds of cattle in Nebraska, North Dakota, and South Dakota. Blood samples were collected before and after the vector season. Samples were tested for antibodies against bluetongue virus by use of a commercially available competitive ELISA. Factors evaluated included descriptors of geographic location and management practices. Trapping of insect vectors was conducted to evaluate vector status on a subset of 57 operations. A multivariable logistic regression model was constructed to evaluate associations. For the full data set, altitude and latitude were associated with risk of having seropositive cattle (an increase in altitude was associated with an increase in risk, and a more northerly location was associated with a decrease in risk of a premise having seropositive cattle). Import of cattle from selected states was associated with an increase in risk of having seropositive cattle. From the subset of herds with data on vector trapping, altitude and latitude were associated with risk of having seropositive cattle, similar to that for the full model. However, commingling with cattle from other herds was associated with a decrease in risk of seropositivity. Findings reported here may be useful in generating additional hypotheses regarding the ecologic characteristics of bluetongue viruses and other vector-borne diseases of livestock. Sentinel surveillance programs are useful for documenting regionalization zones for diseases, which can be beneficial when securing international markets for animals and animal products.

A regional prospective study of the epidemiology of bluetongue virus (BTV) serotypes covering 11 countries in Central America and the Caribbean took place between 1987 and 1992. Active surveillance revealed BTV infection to be endemic in the absence of confirmed indigenous cases of bluetongue. During the 6-year span of the study, over 300 BTV isolations were obtained from cattle and sheep. Results of the earlier years of the study were summarized, and surveillance activities in the concluding months of the study from November 1990 to February 1992 were evaluated. Forty-five BTV isolations were made during this time, 44 from sentinel cattle and 1 from a ram with clinical signs compatible with contagious ecthyma. Virus isolation from potential vectors also was attempted, yielding a further 9 BTV isolates from parous Culicoides insignis and C pusillus, 2 BTV isolates from blood-engorged C filarifer, and 1 epizootic hemorrhagic disease virus type-2 isolate from parous C pusillus. Our extensive network of sentinel herds in the region detected BTV-1 as the predominant serotype in Central America in 1991, after an apparent absence of 1 year in the sentinel animals. Other serotypes in Central America at that time included BTV-3 and BTV-6. In Puerto Rico and the Dominican Republic, BTV-4 became the predominant serotype, without detection of BTV-8 and BTV-17, which were common in recent years of the study. The serotypes found in the Caribbean Basin continued to have marked differences from those in North America. The importance of viewing bluetongue as an infection, the distribution of which is determined principally by ecologic factors, is emphasized.

Inoculation of 53 ewes after 35, 45, 60, or 80 days of gestation with bluetongue virus serotypes 10, 11, 13, or 17, or with epizootic hemorrhagic disease virus serotypes 1 or 2, resulted in overt clinical disease in the 47 ewes inoculated with bluetongue virus but not in the 6 ewes inoculated with epizootic hemorrhagic disease virus. None of the lambs produced by these ewes had developmental defects or any evidence of persistence of viremia.

Results of testing of 19,731 samples from a serologic survey of cattle with bluetongue virus (BTV) infections in Australia were analyzed for association between age, species, or sex and test result. Bivariate analysis indicated that all 3 host factors were associated with test result. After adjusting for confounding caused by the location of each animal in the study (high, moderate, and low BTV prevalence regions), cattle greater than or equal to 4 years old had an odds ratio of 4.33 (95% confidence interval, 3.99, 4.71) for a positive test result, compared with that for cattle < 2 years old. Cattle 2 to 4 years had an odds ratio of 2.28 (2.14, 2.54), compared with cattle < 2 years old. Bos taurus cattle had an odds ratio of 1.76 (1.63, 2.05) of a positive test result, compared with crossbred cattle, and B indicus cattle had an odds ratio of 1.20 (1.09, 1.33), compared with crossbred cattle. Sexually intact (+) male cattle were found to have an odds ratio of 3.13 (2.66, 3.49) for a positive test result, compared with castrated male (-) cattle, and female cattle were found to have an odds ratio of 1.38 (1.29, 1.48), compared with male (-) cattle. Multivariate analysis of BTV testing results was performed, using stepwise logistic regression. The most parsimonious model selected included age, species, and sex factors, and first-order interaction terms between these factors. This model was only able to be fit to data from cattle restricted to the high (> 25%) BTV prevalence region. Odds ratios were found to increase with age for male (-) cattle of all species. Odds ratios were found to be greatest at 2 to 4 years of age for female cattle of all species and for B taurus and crossbred male (+) cattle.

The consequences of inoculation of bluetongue virus (BTV) serotype 11 into 16 susceptible cows either at the time of breeding or at specified stages of pregnancy were studied. The cows were free of BTV or epizootic hemorrhagic disease virus, and none had antibodies to BTV before virus inoculation. A group of 4 cows was mated naturally to a bull reported to shed BTV (CO75B300 strain) in the semen. The bull was suspected of infecting cows at mating with BTV-11, which subsequently transplacentally infected the developing fetuses and induced persistently infected and congenitally malformed progeny. Two groups of 4 pregnant cows were inoculated with an insect-derived strain of BTV-11 (CO75B300), one group by direct deposit into the uterus at estrus, the other, by intradermal and sc administrations. A 90-day fetus was inoculated in utero with virus from the same pool. Four pregnant cows were inoculated with sheep blood-passaged virus of the same BTV-11 strain (CO75B300) by intradermal and sc routes. Three cows were inoculated with BTV-free suspending fluids and ovine erythrocytes by the intrauterine and intradermal-sc routes and were used as in-contact controls.Infection with insect-derived BTV-11 was confirmed in 3 cows of 1 group by virus isolation and by detection of serum antibodies. The 4 cows inoculated with sheep blood suspension of BTV-11 developed viremia and produced antibodies to the virus. None of the cattle had clinical signs of bluetongue, other than 2 cows that had a slight rectal temperature increase on postinoculation day 4.All cows and fetuses that ranged in gestational age from 69 to 217 days appeared grossly normal at necropsy. Antibodies were not detected in fetal blood. Viral antigen was not detected in fetal tissues by inoculation into sheep or by immunofluoerscence, and viral RNA was not detected by use of the polymerase chain reaction. Developmental deformities were not seen in any fetus. The BTV-11 was not transmitted via the bull semen after natural mating. The BTV-11 strain CO75B300, isolated from this bull and passaged either as insect-derived or ovine erythrocyte suspensions, infected 8 cows. However, the virus was not transplacentally transmitted to their fetuses. It was concluded that there was no evidence for congenital BTV-11 infection in this study.