Leptospirosis as a cause of infertility in Uruguayan beef cattle.

Leptospirosis, caused by pathogenic spirochetes of the genus Leptospira spp., is a globally significant zoonotic disease that affects humans and animals. In cattle, leptospirosis is associated not only with overt clinical manifestations but also with reproductive diseases, including infertility. This study assesses the potential correlation between leptospirosis and infertility in Uruguayan beef cattle. A case-control study involved 31 beef herds with no prior history of Leptospira vaccination. In each herd, veterinarians identified 10 non-pregnant (cases) and 25 pregnant cows (controls) using ultrasound, and blood and urine samples were collected from each cow. Serological diagnosis was performed using the Microscopic Agglutination Test (MAT), and quantitative PCR (qPCR) was used to assess Leptospira excretion. Additionally, antibodies against bovine viral diarrhea virus (BVDV) and infectious bovine rhinotracheitis (IBR) were tested. The results demonstrated an association between seropositivity to the Sejroe serogroup (cut-off 1:200) and infertility in cattle (OR=1.31; p-value=0.06). Furthermore, the level of Leptospira excretion (qPCR) in urine was associated with increased infertility risk, with cows excreting over 100 copies per mL of urine having the highest odds of infertility (OR=2.34; p-value<0.01). This study suggests a potential association between leptospirosis and infertility in Uruguayan beef cattle, emphasizing the importance of both serological and molecular diagnostics for assessing reproductive health in cattle herds. Future research should explore the impact of Leptospira serogroups on other reproductive disorders in cattle.

Seroprevalence and herd-level associated factors of pathogenic Leptospira spp. circulating locally in dairy cattle in Uruguay.

Leptospirosis is a zoonotic disease of worldwide importance. In Uruguay, it is endemic in cattle and primarily affects people with occupational exposure to livestock. The aim of this study was to determine the national seroprevalence and associated factors of local pathogen Leptospires in dairy cattle. A cross-sectional study was carried out. Herds were stratified by size (1-50, 51-250, and > 250 cattle), and up to 60 dairy cows per herd were randomly selected. A total of 4269 serum samples from 101 dairy herds were analyzed by microscopic agglutination test (MAT). A two-stage sampling design was used to estimate population seroprevalence of Leptospira spp. In order to determine the factors associated with the disease, herds with at least 1 seropositive animal were considered as case herds. Seroprevalence of Leptospira was 27.80% with a 95% CI [21.06, 34.54] at the animal level and 86.92% with a 95% CI [80.00, 93.75] at the herd level. The serology confirms the predominance of serogroups Sejroe and Pomona in our herd with the presence of incidental leptospires infection, in smaller proportion, but with a wide distribution at farm level. The population size and purchasing replacement of cows on dairy farms were associated with infection at farm level. The serologic studies confirmed that exposure to Leptospira spp. is endemic in our herds, and the spreading over dairy herds. Although the movement of purchased females and the size of the herd were associated with the disease, more studies should be conducted, to better understand the epidemiology of the disease and to highlight the possible risks to public health, especially in rural workers, farmers and veterinarians.

Epidemiological study of neosporosis in Uruguayan dairy herds.

Neosporosis is one of the leading causes of abortion in cattle worldwide, posing a great economic burden on cattle producers. The aim of this study was to determine the national seroprevalence and putative risk factors of Neospora caninum (N. caninum) in dairy cattle in Uruguay. A cross-sectional study was carried out. Herds were stratified by size (1-50, 51-250, and >250 cattle) and up to 60 dairy cows per herd were randomly selected. Four thousand two hundred twenty-three serum samples from 102 dairy herds were analyzed by indirect ELISA test, under the manufacturer's recommendations. In addition, the herdsman was surveyed and a population study was carried out. The in degree data, geographical coordinates, and seroprevalence of bovine viral diarrhea, enzootic bovine leukemia and infectious bovine rhinotracheitis were available for each herd. A sampling design was used to estimate population seroprevalence of N. caninum. In order to determine the factors associated with the disease, herds with an intra-herd seroprevalence over 20% were considered as case herds. Seroprevalence of N. caninum was 22.3% (95% CI: 18.7-25.9 %) and 96.0% (95% CI: 92.1-99.8%) at the animal and herd level, respectively. The number of dogs on the dairy farms were associated with infection levels (OR: 1.43, 95% CI: 1.02 to 2.03). It was concluded that N. caninum is endemic in the country, and is spreading over dairy herds. Although this study showed evidence that the number of dogs were associated with high levels of infection, more studies should be conducted, to better understand the epidemiology of the disease and thus develop efficient control measures.

Analytic Sensitivity of an ELISA Test on Pooled Sera Samples for Detection of Bovine Brucellosis in Eradication Stages in Uruguay.

Bovine brucellosis has been under eradication in Uruguay since 1998. The eradication program includes, among other interventions, individual sera sampling of beef animals at slaughter, and annual serum testing of all dairy cows-accounting for two million samples annually. At a herd prevalence of 0.8%, a pooled-sera sample approach could reduce the economic burden of the surveillance system by reducing the testing and operational costs. Our objective was to evaluate the analytic sensitivity of an indirect ELISA test for Brucella abortus in serum pools. Sixty-two Brucella abortus-positive bovine sera samples (based upon rose bengal and fluorescent polarization assay) were used as the positive control samples. Rose bengal-negative sera from negative farms were used to dilute the positive samples to the desired concentrations. Positive samples were diluted by using 1 ml of positive sera and 1 ml of negative sera (1/2 dilution) up to 1/1,024. Data were analyzed using generalized linear mixed models with a binary outcome (positive or negative), dilution number as a fixed effect, and a random effect for sample ID. Analytic sensitivity was 99.0% [95% confidence interval (CI): 96.3-99.7], 98.3% (95% CI: 93.1-99.6), 97.3% (95% CI: 87.4-99.4) for dilutions 1/2, 1/4, and 1/8, respectively. The analytical sensitivity, however, decreased when diluted to greater proportions. Given the current herd prevalence in Uruguay, it seems plausible that the use of a pooled sample approach could be adopted by policymakers to reduce the cost of the surveillance program and increase the number of samples being tested.

Causes of abortion in dairy cows in Uruguay / Causas de aborto em bovinos de leite no Uruguai

A case series study was conducted to determine the frequency of causes of abortion in dairy cattle in Uruguay. The sample size of 102 cases was composed of 53 fetuses, 35 fetuses with placentas, and 14 placentas without an associated fetus. All cases underwent gross and microscopic pathologic examinations as well as microbiological and serological testing. The etiology was determined in 54 (53%) of cases, 51 of which were caused by infectious agents. Within the observed 102 cases, 30 (29%) were caused by Neospora caninum, six (6%) by Coxiella burnetii and two (2%) by Campylobacter fetus subsp. venerealis. Bovine Parainfluenza-3 virus and Salmonella enterica serovar Newport caused one abortion each. Opportunistic bacteria (Escherichia coli, Streptococcus sp., Staphylococcus sp., Mannheimia sp., Trueperella pyogenes, and Providencia stuartii) were associated with 11 abortions. In two cases the fetal death was attributed to dystocia, and in one case the fetus had a congenital mesothelioma. Bovine viral diarrhea virus (BVDV) infection was identified in three fetuses; two of which were co-infected with and had typical lesions of N. caninum. No lesions were observed in the other fetus infected by BVDV. Leptospira interrogans was identified in one fetus without lesions. Despite the relatively low overall success rate in establishing an etiological diagnosis in cases of abortion in cattle, a systemic workup of bovine abortion is necessary to establish prevention and control strategies. This also facilitates monitoring and surveillance of reproductive diseases in dairy cattle, some of which represent a risk to public health.(AU)

Uma série de casos foi estudada para determinar a frequência de causas do aborto em bovinos leiteiros no Uruguai. A amostra, de 102 casos, foi composta por 53 fetos, 35 fetos com placentas e 14 placentas sem feto associado. Todos os casos foram submetidos a exames patológicos macroscópicos e microscópicos, além de testes microbiológicos e sorológicos. A etiologia foi determinada em 54 (53%) dos casos, 51 dos quais foram causados por agentes infecciosos. Nos 102 casos observados, 30 (29%) foram causados por Neospora caninum, seis (6%) por Coxiella burnetii e dois (2%) por Campylobacter fetus subsp. venerealis. O vírus da Parainfluenza-3 e Salmonella enterica serovar Newport causaram um aborto cada. Bactérias oportunistas (Escherichia coli, Streptococcus sp., Staphylococcus sp., Mannheimia sp., Trueperella pyogenes e Providencia stuartii) foram associadas a 11 abortos. Em dois casos, a morte fetal foi atribuída a distocia e, em um caso, o feto apresentava mesotelioma congênito. A infecção pelo vírus da diarreia viral bovina (BVDV) foi identificada em três fetos; dois dos quais foram co-infectados e apresentavam lesões típicas de N. caninum. Não foram observadas lesões no outro feto infectado pelo BVDV. Leptospira interrogans foi identificada em um feto sem lesões. Apesar da relativamente baixa taxa de sucesso no diagnóstico etiológico nos casos de aborto em bovinos, é necessário o diagnóstico sistemático dos abortos para estabelecer estratégias de prevenção e controle. Isso também facilita o monitoramento e a vigilância de doenças reprodutivas em bovinos leiteiros, algumas das quais representam um risco para a saúde pública.(AU)

Modeling the Accuracy of Two in-vitro Bovine Tuberculosis Tests Using a Bayesian Approach.

Accuracy of new or alternative diagnostic tests is typically estimated in relation to a well-standardized reference test referred to as a gold standard. However, for bovine tuberculosis (bTB), a chronic disease of cattle, affecting animal and public health, no reliable gold standard is available. In this context, latent-class models implemented using a Bayesian approach can help to assess the accuracy of diagnostic tests incorporating previous knowledge on test performance and disease prevalence. In Uruguay, bTB-prevalence has increased in the past decades partially because of the limited accuracy of the diagnostic strategy in place, based on intradermal testing (caudal fold test, CFT, for screening and comparative cervical test, CCT, for confirmation) and slaughter of reactors. Here, we evaluated the performance of two alternative bTB-diagnostic tools, the interferon-gamma assay, IGRA, and the enzyme-linked immunosorbent assay (ELISA), which had never been used in Uruguay in the absence of a gold standard. In order to do so animals from two heavily infected dairy herds and tested with CFT-CCT were also analyzed with the IGRA using two antigens (study 1) and the ELISA (study 2). The accuracy of the IGRA and ELISA was assessed fitting two latent-class models: a two test-one population model (LCA-a) based on the analysis of CFT/CFT-CCT test results and one in-vitro test (IGRA/ELISA), and a one test-one population model (LCA-b) using the IGRA or ELISA information in which the prevalence was modeled using information from the skin tests. Posterior estimates for model LCA-a suggested that IGRA was as sensitive (75-78%) as the CFT and more sensitive than the serial use of CFT-CCT. Its specificity (90-96%) was superior to the one for the CFT and equivalent to the use of CFT-CCT. Estimates from LCA-b models consistently yielded lower posterior Se estimates for the IGRA but similar results for its Sp. Estimates for the Se (52% 95%PPI:44.41-71.28) and the Sp (92% 95%PPI:78.63-98.76) of the ELISA were however similar regardless of the model used. These results suggest that the incorporation of IGRA for detection of bTB in highly infected herds could be a useful tool to improve the sensitivity of the bTB-control in Uruguay.

Leptospira interrogans serogroup Pomona serovar Kennewicki infection in two sheep flocks with acute leptospirosis in Uruguay.

Acute leptospirosis is an infrequent disease in sheep that can cause jaundice, haemolysis, haemoglobinuria, hepatitis and nephritis. In most reports the diagnoses have been made by clinical, pathological or serological evidence without isolation or direct identification of the agent. Here, we report one confirmed and one presumptive outbreak of acute leptospirosis in suckling lambs from two unrelated sheep farms in Uruguay with mortalities of 9/60 (15%) and 9/163 (5.5%) lambs. Both outbreaks occurred in Sep-Oct 2017 after heavy rainfall and flooding events. The main gross and histologic pathological findings in two autopsied lambs, one from each farm, included severe diffuse jaundice, haemoglobinuria, acute necrotizing hepatitis with cholestasis and interstitial nephritis. Leptospira interrogans serogroup Pomona serovar Kennewicki was isolated from sheep in both flocks and the same genotype was identified directly in clinical samples from infected animals, including one of the deceased lambs subjected to autopsy, by amplification and partial sequencing of rrs and secY genes. This serovar has recently been identified in infected cattle and humans in Uruguay. The impact of Leptospira spp. infection in ovine health, and the epidemiologic role of sheep as reservoirs of leptospirosis for humans and animals need further investigation.

Isolation of pathogenic Leptospira strains from naturally infected cattle in Uruguay reveals high serovar diversity, and uncovers a relevant risk for human leptospirosis.

Leptospirosis is a neglected zoonosis with worldwide distribution. The causative agents are spirochete bacteria of the Leptospira genus, displaying huge diversity of serovars, the identity of which is critical for effective diagnosis and vaccination purposes. Among many other mammalian species, Leptospira infects cattle, eliciting acute signs in calves, and chronic disease in adult animals often leading to abortions. In South America, and including in Uruguay, beef and dairy export are leading sources of national income. Despite the importance of bovine health, food safety, and bovine-related dissemination of leptospirosis to humans, extremely limited information is available as to the identity of Leptospira species and serovars infecting cattle in Uruguay and the South American subcontinent. Here we report a multicentric 3-year study resulting in the isolation and detailed characterization of 40 strains of Leptospira spp. obtained from infected cattle. Combined serologic and molecular typing identified these isolates as L. interrogans serogroup Pomona serovar Kennewicki (20 strains), L. interrogans serogroup Canicola serovar Canicola (1 strain), L. borgpetersenii serogroup Sejroe serovar Hardjo (10 strains) and L. noguchii (9 strains). The latter showed remarkable phenotypic and genetic variability, belonging to 6 distinct serogroups, including 3 that did not react with a large panel of reference serogrouping antisera. Approximately 20% of cattle sampled in the field were found to be shedding pathogenic Leptospira in their urine, uncovering a threat for public health that is being largely neglected. The two L. interrogans serovars that we isolated from cattle displayed identical genetic signatures to those of human isolates that had previously been obtained from leptospirosis patients. This report of local Leptospira strains shall improve diagnostic tools and the understanding of leptospirosis epidemiology in South America. These strains could also be used as new components within bacterin vaccines to protect against the pathogenic Leptospira strains that are actually circulating, a direct measure to reduce the risk of human leptospirosis.