Seropositive Cows Research Articles

Is it safe to use ovarian follicular fluid from cows seropositive for brucellosis in the production of invitro embryos?

Animal reproduction biotechniques are important tools for the technological advancement of livestock, as they allow the selection of the reproductive potential of superior quality females and males; however, infectious diseases that have a predilection for the reproductive system can be a hindrance for the use of these technologies. Therefore, the present study aimed to detect Brucella spp. in the ovarian follicular fluid of brucellosis-positive bovine cows. A total of 47 bovine ovarian follicular fluid aspirates from cows, positive in tests for brucellosis and from Brucella-positive herd, were submitted to PCR. The primers used in the PCR were specific to the genus Brucella (bcsp31 gene). All 47 bovine aspirates were negative for Brucella spp. 0.00% (95% CI: 0.00-4.00%). Our results demonstrated that Brucella spp. was absent in the ovarian follicular fluid from seropositive cows, which indicates that Brucella spp.-infected cows could be used for reproductive biotechnologies carried out with follicular aspirates. Future studies are needed to more precisely evaluate the feasibility and safety of using these oocytes from brucellosis-seropositive cows to transfer embryos to heifers/cows not infected by Brucella, aiming to produce calves free of the infection.

Serological Profile for Major Respiratory Viruses in Unvaccinated Cows from High-Yielding Dairy Herds.

This study aims to determine the serological profile of high-yielding dairy cows for four main viruses (bovine alphaherpesvirus 1 (BoAHV1), bovine viral diarrhea virus (BVDV), bovine parainfluenza virus 3 (BPIV3), and bovine respiratory syncytial virus (BRSV)) related to bovine respiratory disease (BRD) in cattle herds worldwide. In this survey, 497 blood serum samples were collected from non-vaccinated dairy cows without clinical respiratory signs in 39 herds in the central-eastern mesoregion of Paraná State, South Brazil. The presence of neutralizing antibodies was determined by virus neutralization (VN) tests. VN antibodies against BoAHV1, BVDV, BPIV3, and BRSV were detected in 355 (71.4%), 280 (56.3%), 481 (96.8%), and 315 (63.4%) serum samples, respectively. The frequencies of seropositive herds for BoAHV1, BVDV, BPIV3, and BRSV were 79.5 (n = 31), 82.0 (n = 32), 100 (n = 39), and 84.6% (n = 33), respectively. The frequencies of seropositive cows varied according to the type of herd management and the number of cows in the herd. The detection of VN antibodies in unvaccinated dairy cattle herds demonstrated the endemic circulation of the four viruses in the herds evaluated. For BRD prevention, it is recommended to implement a vaccination program for cows that provides passive immunity in calves and active immunity in cows.

Detection of Toxoplasma gondii DNA in milk of dairy cows from southern Brazil.

Consumption of unpasteurized cow's milk may be a transmission route for some pathogenic microorganisms, but there is little information about the risk of Toxoplasma gondii infection. Blood and milk samples were collected in a paired and random fashion from 106 dairy cows and bulk-tank milk samples were also collected from each of the six farms, in southern Brazil. Serum anti-T.gondii antibodies (IgG) were detected by an indirect fluorescent antibody test (IFAT) with a cutoff point of 1:64. Nested PCR targeting the ITS1 was performed on milk samples to detect the Sarcocystidae family, confirmed to be T.gondii by Sanger sequencing. The occurrence of anti-T.gondii antibodies in the herds was 14.1%, (15/106) with seropositive cows in all herds. Antibody titers in positive samples ranged from 64 to 128. T.gondii DNA was detected in 2.8% (03/106) of the milk samples. The ITS1 sequences generated in this study were ON809793 - ON809794 and the sequencing revealed 98-100% identity with T. gondii DNA sequences deposited in GenBank. All cows PCR positive for T.gondii in milk were negative for IgG antibodies in serum, suggesting that naturally infected cows may shed T. gondii in milk in the acute phase of infection. The results of this study demonstrate that T. gondii DNA may be detected in raw cow's milk, so the potential risks of lactogenic infection should be considered. The presence of T. gondii DNA in milk does not confirm that the protozoa are viable and infective, and further investigations into the role of cow's milk in the epidemiology of toxoplasmosis are needed.

Investigation of brucellosis in culled cows and professionals at El-Harrach’s slaughterhouse in Algeria

¶In Algeria, despite control program (test/slaughter) applied since 1995, animal and human brucellosis incidence remains high over the last years. Our ¶Ou survey was carried out with, an aim of determining the insufficiencies of the applied prophylaxis plan, and assessing brucellosis hazard on professionals of slaughterhouse. ¶On top of the 31 visits performed at the slaughterhouse of El-Harrach, blood sera and data associated with: age, identification statute, race, type of slaughtering, pregnancy, and metritis presence were collected from 351 cull-cows¶. ¶From the cooperative professionals of the same establishment, 31 sera in conjunction with the socio-epidemiological data were sampled. ¶All sera were analyzed with Rose Bengal Plat Agglutination Test (RBPAT). ¶The obtained results showed 0% prevalence in the slaughterhouse workers, and a global rate of 4.56 % RBPAT positivity in the cull-cows, within 5.55% prevalence in the pregnant ones. A rate of 50% (28.6 to 83.3%) daily frequency appearance of a sero-positive cow at the slaughterhouse of El-Harrach was established. ¶In addition, 4.01% slaughtered cows in the framework of healthiness slaughtering, were positive. ¶While, no difference in prevalence related to race, neither to identification statute, nor for age, were recorded. ¶Indeed, one can expect that the uncontrolled movement of animals can evoke a substantial factor limiting the effectiveness of brucellosis control program.

Shedding rate of Brucella spp. in the milk of seropositive and seronegative dairy cattle

Brucellosis in cattle herds has caused severe economic losses in many regions worldwide. A cross-sectional study was performed to investigate the presence of Brucella spp. in industrial dairy cattle farms in Iran. For this purpose, 935 blood and 935 milk samples were randomly collected from industrial dairy cattle farms in Iran's Alborz and Tehran provinces. Blood and milk samples were collected on the same day from each cow. Serological, bacteriological, and molecular characterization of Brucella isolates were performed using standard methods. Our results revealed the seroprevalence of brucellosis in dairy cattle farms in the Alborz and Tehran provinces, reaching 19.8%, 6.7%, 5.1%, 14.1%, and 13.1% using the Rose Bengal plate test (RBPT), serum agglutination test (SAT), 2-mercaptoethanol test (2-ME), indirect enzyme-linked immunosorbent assay (i-ELISA) and milk ring test (MRT), respectively. Furthermore, the results of bacterial culture and PCR analyses showed the presence of Brucella abortus among dairy cattle in the Alborz province and Brucella melitensis and B. abortus among dairy cattle in the Tehran province. Moreover, statistical analysis with Cohen's Kappa has highlighted the near-perfect agreement between RBPT and i-ELISA (k = 0.86). In contrast, substantial agreement was shown between RBPT and SAT performance (k = 0.70) and moderate agreement between RBPT and 2-ME (k = 0.67). The findings of this investigation showed shedding of Brucella in the milk of seropositive cows, which is a serious problem involving the maintenance and further spread of Brucella infection on the farm. Therefore, for brucellosis detection or eradication in dairy cattle farms, bacteriological and serological tests of milk samples should be performed along with blood analysis to inhibit the uncontrolled spread of the disease in animals and humans.

Serum and Milk Levels of Antibodies to Bovine Viral Diarrhoea Virus, Bovine Herpesvirus-1 and -4, and Circulation of Different Bovine Herpesvirus-4 Genotypes in Dairy Cattle with Clinical Mastitis.

Bovine viral diarrhoea virus (BVDV) and bovine herpesvirus (BoHV)-1 and -4 are important causes of respiratory diseases and reproductive disorders of dairy cattle worldwide. Investigation of BVDV and BoHV-1 and -4 antibody levels in the serum and milk of dairy cattle in a group with clinical mastitis and a healthy group was undertaken using an indirect ELISA, and identification of the BoHV-4 genotypes in clinical mastitis cases was attempted by PCR and sequencing. Antibodies specific to BVDV, BoHV-1 and BoHV-4 were detected in the serum and milk of all dairy cattle with clinical mastitis. The cut-off values for BVDV and BoHV-1 in the sera and milk were extremely high in both healthy and mastitic animals. However, BoHV-4 antibodies were detected only in the clinically mastitic cattle, and BoHV-4 levels were higher in milk than in sera among these animals. Genotypes I and II of BoHV-4 were detected in the milk samples of four seropositive cows with clinical mastitis from the same herd. The results of this investigation demonstrate that clinical mastitis cases in the same herd may have aetiology in different BoHV-4 genotypes.

Neospora in UK cattle

Neosporosis was first identified in the 1980s and has since become well established as an endemic disease of cattle and a significant cause of abortion. Neospora caninum is an obligate, intracellular, protozoal parasite that relies on intermediate and definitive hosts to complete its lifecycle. Neospora infection leaves cattle three to thirteen times more likely to abort than a healthy animal. Calves may be born alive but be latently infected owith Neospora or have obvious defects. Disease transmission can be horizontal, when cattle ingest feed contaminated with infected dog faeces that results in latency or exogenous transplacental infection, or vertical when latent infection recrudesces, producing a congenitally infected calf. Testing involves serology or direct testing if abortion material is available. Serology can provide a presumptive diagnosis when carried out near in time to an abortion event. Eradicating the disease involves identifying infected cows and infected family lines through serology and removing these individuals from the herd. This approach takes time because seropositive cows do not always return positive tests, and in a herd with a high seroprevalence it is not economically viable to remove large numbers of animals.

Epidemiological evaluation of Neospora caninum in dairy animals

The purpose of the study is to explore the epidemiological information concerning the spread of neosporosis in the Moscow region (Russian Federation) and Almaty region (Republic of Kazakhstan). The study is conducted in 2019 in the Moscow region (Russian Federation) and Almaty region (Republic of Kazakhstan). The study sample includes 800 cows (400 animals at each of the two locations), 100 at each of the 4 cattle farms in the Moscow region and 100 at each of the 4 cattle farms in the Almaty region. There were more seropositive cows on the rest of farms as compared to farm No. 1: 1.9 times more on farm No. 2 (p ≤ 0.01), 2.4 times more on farm No. 3 (p ≤ 0.001), and almost 4 times more on farm No. 4 (p ≤ 0.0001). In terms of the abortion rates, the maximum difference between the farms was 5-fold in the Moscow region (p ≤ 0.0001) and 3-fold in the Almaty region (p ≤ 0.001). Positive correlations exist between all the studied parameters (the proportion of seropositive animals, the proportion of seroprevalent animals, the abortion rate, and the stillbirth rate). The results of the study are mainly extremely valuable for the global economy, because the Republic of Kazakhstan and the Russian Federation occupy an important place in the export of meat and dairy products.

Soroprevalência em nível de rebanho e fatores associados para Toxoplasma gondii em vacas no estado da Paraíba, Nordeste do Brasil

We aimed to determine the herd and animal levels seroprevalence and associated factors for Toxoplasma gondii infections in cattle from the state of Paraíba, Northeastern Brazil. Herds (n = 434) and cows aged ≥ 24 months (n = 1,895) were randomly selected, and serum samples were tested with the immunofluorescence antibody test (IFAT) using as cutoff of 64. Of the 434 farms investigated, 197 had at least one seropositive cow (prevalence of 49.0%; 95% CI = 44.3%-53.8%), and the prevalence at animal level was 18.0% (95% CI = 5.3%-21.1%). The antibody titers ranged from 64 to 1024, with the most frequent titers being 64 (10.8%) and 128 (3.7%). The risk factors identified were property located in Sertão region (odds ratio [OR] = 3.07), property located in Agreste/Zona da Mata regions (OR = 2.00), animal purchasing (OR = 2.68), herd size of 34-111 animals (OR = 2.91) and herd size > 111 animals (OR = 6.97). The results suggest the wide distribution of T. gondii infections in cattle throughout the state of Paraíba, and the risk factors identified are not possible to correct.

Screening for Coxiella Burnetii in Dairy Cattle Herds in Poland.

The intracellular bacterium Coxiella burnetii is the aetiological agent of Q fever, a zoonosis affecting many animal species worldwide. Cattle and small ruminants are considered the major reservoirs of the bacteria and they shed it through multiple routes. A total of 2,180 sera samples from 801 cattle herds in all Polish voivodeships were tested by ELISA for the presence of specific antibodies. Milk samples were obtained from seropositive cows in 133 herds as part of a separate study. The milk samples were examined by ELISA and real-time PCR tests. Seroprevalence at the animal level was 7.06% and true positive seroprevalence was 6.0% (95% confidence interval (CI) 1.1-9.4). Seroprevalence at the herd level was estimated at 11.1% and true positive seroprevalence was 10.5% (95% CI 3.2-15.8). Shedding of the pathogen in milk was detected by real-time PCR in 33 out of 133 tested herds (24.81%, 95% CI 17.74-33.04%) and the presence of C. burnetii antibodies was confirmed in 85 of them (63.9%, 95% CI 55.13-72.05%). The highest level of conformity between ELISA and real-time PCR results was obtained for bulk tank milk samples. Coxiella burnetii infections are quite common in cattle herds across the country, which emphasises the crucial roles of surveillance and adequate biosecurity measures in the prevention and limitation of Q fever spread in Poland.

Presence of antibodies against Mycobacterium avium subspecies paratuberculosis in Brazilian high-producing dairy herds.

This study aimed to determine the presence of antibodies against Mycobacterium avium subspecies paratuberculosis (MAP) in high-producing dairy cows, the presence of the pathogen in the feces, and the risk factors associated with the disease. Blood and fecal samples were collected from 708 dairy cows over 2years from 54 herds located in five municipalities of Paraná, Brazil. The serum samples were evaluated for the presence of antibodies against MAP using enzyme-linked immunosorbent assay (ELISA). Fecal samples from 100 cows (69 seropositive and 31 seronegative) were assessed using real-time PCR (qPCR) for IS900 of MAP. The herd prevalence of antibodies against MAP was 61.1% (33/54; 95% CI 46.88-74.08), ranging from 12.5 to 80% across the municipalities, and the prevalence in the animals was 9.8% (69/708; 95% CI 7.77-12.15); it ranged from 0 to 87.5% per herd. Only one of the 69 (1.45%) fecal samples from the seropositive cows was positive for the qPCR. The factors associated with the occurrence of paratuberculosis in herds were the use of compost barn system and the type of bed, whereas only the type of bed was associated with the infection of cows. The only risk factor (OR = 2.45; 95% CI 1.03-5.85) associated with the occurrence of paratuberculosis was the introduction of animals purchased from other dairy farms. The prevalence of active infection was low; however, our results demonstrate the presence of MAP in high-producing dairy herds in Paraná state, Brazil.

Parasitemia and Associated Immune Response in Pregnant and Non-Pregnant Beef Cows Naturally Infected With Neospora caninum.

The aim of this longitudinal study was to characterize the parasitemia of Neospora caninum and the associated immunological parameters in naturally infected beef cows for 10 months. The following groups were established: Neospora caninum seropositive pregnant cows (+Preg, n = 7), seropositive non-pregnant cows (+Npreg, n = 7), seronegative pregnant cows (−Preg, n = 4), and seronegative non-pregnant cows (−Npreg, n = 4). Several samples were obtained for absolute and relative leukocyte counting, cytokines IL-10, IL-12, α-TNF, and γ-IFN quantification, specific IgG, IgG1, and IgG2 and avidity and N. caninum DNA molecular detection and quantification. The +Preg group had a higher frequency and concentration of N. caninum DNA in PBMC in the last third of pregnancy compared to +Npreg (p <0.05), with 22 and 8% of detection, respectively. Parasitemia correlated positively with IgG titers and negatively with IgG1/IgG2 ratio (p <0.05). On day 222 of the assay, the +Preg group had the lowest total leukocyte counting (p <0.05). The +Preg group had a higher concentration of IgG and higher avidity in the last third of gestation compared to +Npreg (p <0.05). Avidity correlated with total IgG and IgG2 (p <0.05). All +Preg cows gave birth to clinically healthy but seropositive calves before colostrum intake, therefore, the congenital transmission was 100% efficient. Only a complete N. caninum genotype from a placenta and a partial genotype from cow #3 of the group +Preg were achieved by multilocus microsatellite analysis. Overall, N. caninum parasitemia is frequent in seropositive beef cows during the last third of gestation. This correlates with higher antibody levels and a decrease in total leukocyte counting. The precise timing of the parasitemia may be used for diagnosis purposes and/or for design strategies to avoid vertical transmission. Further studies are needed to identify the immune molecular mechanisms that favor parasitemia during gestation in chronically infected cattle.

Impact of Endemic Besnoitiosis on the Performance of a Dairy Cattle Herd.

Simple SummaryBovine besnoitiosis, caused by the Apicomplexa parasite Besnoitia besnoiti, has been emerging in Europe as a disease of economic concern to the cattle industry. It is a chronic and debilitating disease mostly reported in beef cattle. However, in Europe, bovine besnoitiosis is increasingly common in dairy cattle; therefore, there is a need to assess the impact of this disease on milk production. To study the effect of B. besnoiti infection on dairy production and reproduction, a serological screening was performed on a dairy herd in an endemic area. The results showed that the herd was endemically infected, with high seroprevalence and low clinical prevalence, and the time on herd represented a risk factor to acquire the infection. Seropositive animals and cows with chronic skin lesions revealed higher milk somatic cell counts, and no negative impact on reproductive performance was found.This study aimed to assess the effect of Besnoitia besnoiti infection on the reproductive and productive performance of a dairy cattle herd. A serological screening was performed by indirect fluorescent antibody test (IFAT) on every animal aged over one year (n = 262). Subsequently, 211 animals were clinically examined, with 96 of those being screened for detection of sclerocysts. The overall seroprevalence was 62.9% (CI95%: 56.1–69.5%). On clinical examination, 7.6% (16/211) of the animals presented chronic skin lesions, and 47.9% (46/96) had sclerocysts. Multivariate logistic regression showed that the time on herd represented a risk factor, and the odds of acquiring the infection increased 1.683× per additional year on herd, ranging from less than a year to 8 years. Seropositivity and the presence of sclerocysts revealed an association with a higher milk somatic cell count, which may have a considerable economic impact on dairy production. Regarding reproductive indicators, no negative impact could be associated with clinical besnoitiosis or positive serological results. In conclusion, our study highlights the need to thoroughly evaluate the economic impact of this emerging disease in dairy herd production to help with decision making at both herd and regional levels, particularly in endemic areas.

The Utility of Serological Analysis for Neospora caninum Infection in Dairy Cattle Farms Management: Serological Investigation and Evaluation of the Effects on Reproductive and Productive Performances in Two Study Herds in Northern Italy.

Simple SummaryAmong infectious agents triggering reproductive disorders in cattle, Neospora caninum, an obligate intracellular protozoan parasite, is a major cause of abortion in cows worldwide. A serosurvey was designed in two cattle herds, both located in northern Italy and with similar reproductive management features, recruited as a case study. The clinical outcome and the effects on herd performances were evaluated in association with the seropositivity to N. caninum. This study showed that the integration of serological analysis results for N. caninum, the reproductive and productive parameters, and the information on herd performances, could give indications for the application of control strategies.Neospora caninum is recognized as a major cause of abortion in cattle, determining economic losses, particularly in dairy industries. To evaluate the impact of neosporosis on herd efficiency, and to understand how the serological analysis for N. caninum is explicative of the farm picture, an epidemiological study was designed in two dairy farms recruited as a case study. Blood samples were collected from 540 animals, including cows and heifers over 12 months, and analyzed by an indirect immunofluorescent antibody test with subsequent antibody titration. Overall, 94 animals (17.4%) were identified as positive to N. caninum antibodies (15.5% and 18.5% in Farm 1 and Farm 2), with differences between the farms concerning the antibody titers (Chi-square, p-value = 0.04), particularly in cows (Chi-square, p-value = 0.018). Consequently, a different pattern of abortion episodes was depicted in the two investigated farms. Considering reproductive and productive performances, the number of inseminations necessary to make an animal pregnant was higher in seropositive cows (2.4 and 2.9 in Farm 1 and 2, respectively) than in seronegative ones (2.1 and 2.4 in Farm 1 and 2, respectively). Similarly, particularly in Farm 1, the number of days in milk of not-pregnant cows was higher in seropositive (167.7) than seronegative animals (133.4). Moreover, although the association between N. caninum infection and milk production is still unclear, both the daily milk production and the 305-mature equivalent milk yield were lower in seropositive (31.02 and 11,838.94) than seronegative cows (33.59 and 12,274.88) in Farm 1; an opposite pattern was otherwise depicted in Farm 2. The study showed that even if N. caninum circulated equally in the two herds, the dynamics of the parasite infection and its outcome may be different, also depending on the specific situation of the farm. In this way, the integration of serological analysis for N. caninum, the reproductive and productive parameters, and the information on herd performances, could give specific indications for the application of control strategies.

A two-year serological study of bovine viral diarrhea virus, bovine alphaherpesvirus 1 and bovine parainfluenza virus type 3 in Qazvin dairy cattle farms, Northwestern of Iran

Infections with bovine viral diarrhea virus (BVDV), bovine alphaherpesvirus 1 (BoHV-1), and bovine parainfluenza virus type 3 (BPIV-3) cause diseases in cattle with serious economic consequences worldwide. The objective of the present study was to determine of herd-level and animal-level BVDV, BoHV-1, and BPIV-3 seroprevalence, and evaluate some of the associated risk factors on farms in Qazvin province, Northwestern Iran. A total of 1036 cattle in 16 herds were randomly selected, and their serum samples were tested to detect antibodies to these viruses in a cross-sectional study over 2 years. The results showed the seroprevalence of BVDV, BoHV-1, and BPIV-3 was 100%, 56.3%, and 100% at herd-level and 55.1%, 5.1%, and 95.2% at animal-level, respectively. Statistical analysis revealed that the farm was a strong risk factor for all the studied viruses, while the year was determined as a risk factor for only BVDV (P&lt;0.001). The seroprevalence of BVDV and BPIV-3 was significantly (P&lt;0.01) affected by season. The proportion of seropositive cows increased with age for BVDV and BoHV-1 (P&lt;0.001). Concurrent infection was the highest in mixed infections with BVDV and BPIV-3 (53.2%), and there was a positive correlation between BVDV and BoHV-1 seropositivity (R2= 0.106, P&lt;0.001). The present study shows that infections of BVDV and BPIV-3 are common in cattle in Northwestern Iran and which implies the need to implement control programs to reduce the risk of the spread of these viruses.

Identification and molecular characterization of Brucella abortus and Brucella melitensis isolated from milk in cattle in Azerbaijan

BackgroundAzerbaijan currently ranks thirteenth in global incidence of human brucellosis, with an estimated annual incidence through 2000 at over 50 cases per million. Brucella melitensis has been isolated from patients and is thought to have been acquired through contact with small ruminants or as a foodborne infection. To reduce the burden of human brucellosis, the Azerbaijani government began in 2002, a nationwide vaccination control campaign in small ruminants. There is serological evidence of bovine brucellosis (presumably due to Brucella abortus) in Azerbaijan, but no prevalence estimates were available when this study started in March 2017. The aim of this study was to isolate and identify Brucella spp. from cow milk in the Ganja region, where brucellosis takes a heavy toll on humans and livestock.ResultsBlood and milk samples were collected from cows (n = 1075) in early lactation (up to 90-days) in farms that had a history of previous positive serological results and abortions. Twenty-two out of 57 milk samples collected from seropositive cows, showed growth on Farrell’s media, when incubated with 5% CO2. Eight additional milk samples showed growth in the absence of CO2. The classical biotyping classified them as Brucella abortus (22) and Brucella melitensis (8). RT-PCR confirmed that strains belonged to the genus Brucella. MLVA profiles were obtained for DNA extracted from two B. abortus and six B. melitensis strains. While the B. abortus genetic profile was described in the MLVA database, matching the profile of B. abortus strains isolated in East Europe, Central Asia and China, we found a new genotype for the B. melitensis strains isolated in Azerbaijan, clustering with strains belonging to the American clade, rarely identified in the region.ConclusionDespite the implementation of the vaccination program in small ruminants, our results suggest that spill-over events of B. melitensis from small ruminants to cattle have occurred. However, cattle are likely to be primarily infected with B. abortus, which warranted the implementation of a bovine brucellosis program. Such a program started in fall 2017. In the Ganja region, cattle should be considered as a potential source of B. abortus and B. melitensis for humans.

Phenotypic Selection of Dairy Cattle Infected with Bovine Leukemia Virus Demonstrates Immunogenetic Resilience through NGS-Based Genotyping of BoLA MHC Class II Genes.

Characterization of the bovine leukocyte antigen (BoLA) DRB3 gene has shown that specific alleles associate with susceptibility or resilience to the progression of bovine leukemia virus (BLV), measured by proviral load (PVL). Through surveillance of multi-farm BLV eradication field trials, we observed differential phenotypes within seropositive cows that persist from months to years. We sought to develop a multiplex next-generation sequencing workflow (NGS-SBT) capable of genotyping 384 samples per run to assess the relationship between BLV phenotype and two BoLA genes. We utilized longitudinal results from milk ELISA screening and subsequent blood collections on seropositive cows for PVL determination using a novel BLV proviral load multiplex qPCR assay to phenotype the cows. Repeated diagnostic observations defined two distinct phenotypes in our study population, ELISA-positive cows that do not harbor detectable levels of provirus and those who do have persistent proviral loads. In total, 565 cows from nine Midwest dairy farms were selected for NGS-SBT, with 558 cows: 168 BLV susceptible (ELISA-positive/PVL-positive) and 390 BLV resilient (ELISA-positive/PVL-negative) successfully genotyped. Three BoLA-DRB3 alleles, including one novel allele, were shown to associate with disease resilience, *009:02, *044:01, and *048:02 were found at rates of 97.5%, 86.5%, and 90.3%, respectively, within the phenotypically resilient population. Alternatively, DRB3*015:01 and *027:03, both known to associate with disease progression, were found at rates of 81.1% and 92.3%, respectively, within the susceptible population. This study helps solidify the immunogenetic relationship between BoLA-DRB3 alleles and BLV infection status of these two phenotypic groupings of US dairy cattle.

Seroprevalence to common infectious abortifacient and infertility causing agents in the dairy herds of India.

Information on the prevalence of infectious agents in dairy farms forms the basis for formulating a suitable control strategy; especially in endemic situations. A cross-sectional study was undertaken to determine the prevalence of six economically important bovine diseases, causing reproductive disorders including bovine abortion in organized dairy herds in India. A total of 1,075 animals (cattle and buffaloes) from 09 dairy farms were screened by ELISA tests. Bovine viral diarrhoea (BVD) was the most prevalent (56.5%) disease followed by infectious bovine rhinotracheitis (IBR) (45.4%). Prevalence of Q-fever (5.4%) and neosporosis (6.1%) were less on the farms. Although 16.3% of the samples turned positive for brucellosis, the contribution of calf-hood vaccination (B. abortus S19 vaccine) to the prevalence of antibodies cannot be ruled out. The overall prevalence of bovine anaplasmosis, known to cause sporadic abortions in dairy herds, was 34.1% in the 9 farms with a prevalence of less than 20% in 5 farms. Infection of multiple abortifacient (seroprevalence to more than two pathogens) was recorded in 56.8% of animals. A very strong association was observed between BVD and brucellosis (Odds ratio 14.2; P<0.001). Further, a positive association was also seen between seroprevalence of IBR and anaplasmosis, and neosporosis and Q fever (P<0.05). Viral diseases were found to be more common in the dairy herds than bacterial and protozoan diseases. Increased susceptibility of IBR seropositive cows to other bacterial and viral infections was observed.

Molecular detection and phylogenetic analysis of Neospora caninum in various hosts from Iran

Different N. caninum strains with distinct biological characteristics and pathogenicity have been introduced from various areas of the world. This study aimed to detect Neospora-infection in different hosts from Iran using the nested-Polymerase Chain Reaction (PCR) technique and to perform the phylogenetic analysis of N. caninum isolates using the Nc-5 gene. Different species of parasite hosts (cattle, dogs, rodents, cats, and birds) from the dairy farms with history of abortion with intensive breeding systems in Hamedan Province, Iran were included for sampling. The stillbirth fetuses of cattle, sheep, and goats were also collected from the slaughterhouse. The nested-PCR technique was used to identify the Neospora-DNA in the aborted fetal materials. The PCR products of the positive samples were sequenced and compared with other isolates in GenBank. Antibodies to N. caninum was found in 19.1% of dogs. Also, 11.4% of dogs were positive for Neospora-like oocysts using microscopy method. The molecular prevalence of N. caninum was 10.6% in dogs, 13.6% in cattle, 35.4% in the aborted fetuses of cows, 31.9% in rodents, 7.6% in pigeons, 1.9% in sparrows, 5.1% in cats, and 7% in the semen sample of bulls. A sampled coyote was positive while the stillbirth fetuses of sheep and goats were negative. The multiple sequence alignments indicated that 34 sequences of N. caninum bore a 94.9-99.3% similarity to each other. Hence, these sequences were classified into two major clusters (clusters A and B) and seven sub-groups (isolates 1-7). The sequence of coyote had a high similarity to the sequences derived from dogs and cows. Also, root diagram showed the isolates of current study and previous reported from Iran have a similar origin. This study provided a phylogenetic analysis of Neospora-infection in different hosts, and for the first time, explored Neospora-infection in coyotes from Iran. Different isolates were detected in the studied population in the region. Comparison of the sequences showed the farm dogs play an important role in the life cycle and distribution of the parasite; so the relationship of dogs with the herd and their access to aborted materials should be restricted. Also, regular control of rodents' population and culling of seropositive cows with a history of abortion is essential for cutting the chain of transmission.

Seroprevalence of bovine vaccinia in cows and its correlation with the productive profile of affected farms in Distrito Federal, Brazil.

Bovine vaccinia (BV) is an infectious disease caused by Vaccinia virus (VACV) characterized by vesicular and exanthematic lesions, mainly in cattle. Although BV has been described in some Brazilian regions in the last decades, official information regarding the current prevalence in bovine herds of Midwestern Brazil is lacking. Thus, the current study aimed to estimate the seroprevalence and risk factors associated with BV in cattle in the Distrito Federal (DF), Brazil. Sera of 312 cows of 64 herds were tested by virus-neutralizing test for VACV antibodies. Herd and animal seroprevalence were estimated to be 33.3% (CI 95%: 18.2-48.3%) and 10.6% (CI 95%: 1.0-20.2%), respectively. Seropositive cows were detected in dairy, beef, and mixed-purpose farms. The results of an epidemiological questionnaire showed that no risk factor analyzed was positively associated with seropositivity to VACV. There was no significant association between type of milking (manual/mechanic) and seropositivity to VACV; however, most seropositive cows were present in farms with high daily milk production and high number of lactating and adult cows. Our results indicate that VACV circulates in many regions of DF with considerable prevalence in dairy cows. Control measures to restrict VACV circulation and consequences of the infection may be advisable.