Cross-sectional survey for sheep leptospirosis in the northeast region of Brazil.

Sheep farming has shown expressive growth, even in the Northeast region of Brazil, with rainfall below expectations in the last decade. This growth has been observed not only in the number of herds, but also in the number of properties involved in sheep farming. Although expressive, the production of small ruminants still presents low levels of performance, mainly due to the occurrence of diseases, which constitute an obstacle to this activity. The aim of this study was to determine the factors associated with the seroprevalence of leptospirosis in sheep in Northeastern Brazil. Blood samples were collected from 3438 sheep in 190 herds from 48 municipalities, belonging to the states of Ceará, Paraíba, Piauí, Rio Grande do Norte and Sergipe. Serological diagnosis was performed using the microscopic agglutination test (MAT). In 76.3 % (95 % CI = 69.79-81.81) of the herds there was at least one animal seropositive for at least one of the Leptospira spp. serogroups and 14.94 % (95 %CI = 13.8-16.18) of the animals were seropositive. The state of Paraíba had the highest herd (95.7 %) and animal (26.6 %) prevalences, followed by Ceará (93.9 % and 23.7 %, respectively). The most frequent serogroups were Autumnalis (18.68 %), Australis (17.9 %) and Pyrogenes (15.18 %). There was no association between the studied variables and the herd-level prevalence of leptospirosis. High herd and animal-level seroprevalences was found for sheep leptospirosis in the semiarid region of Northeastern Brazil, and there is a need to implement control and prevention measures, such as quarantine, serology, vaccination and adequate treatment, in addition to the introduction of proven healthy animals, mainly in the states of Paraíba and Ceará, aiming to reduce the prevalence of disease.

Use of serological and molecular techniques for detection of Leptospira sp. carrier sheep under semiarid conditions and the importance of genital transmission route.

It is possible that there are peculiarities in the epidemiology of leptospirosis in regions with a semiarid climate, where the environment is often adverse, allowing the occurrence of alternative transmission routes. The objective of the work was to generate contributions to the diagnosis and epidemiology of Leptospira sp. infection in sheep reared in semiarid conditions, using serological, molecular and microbiological techniques for diagnosis in dry and rainy seasons. Samples of blood, vaginal fluid, urine, bladder, kidney, uterus, uterine tube, ovary and placenta were collected from 104 sheep (52 animals per season - dry and rainy) slaughtered in the Brazilian semiarid. Diagnostic tests performed were microscopic agglutination test (MAT), polymerase chain reaction (PCR) and bacterial isolation. Anti-Leptospira sp. antibodies were found in 26 (25%) of the animals analyzed by MAT at 1:50 dilution (cut-off 50), while 69 (66.3%) animals had at least one organ/fluid with the presence of Leptospira sp. DNA. Overall, PCR was performed on 758 fragments of organs/fluids from the genital and urinary tracts, and 519 (68.5%) samples tested positive. PCR-positivity was statistically different in dry (46.2%) and rainy (11.5%) seasons for vaginal fluid. It was possible to perform the DNA sequencing in nine samples with 99% similarity to L. interrogans and recovery of viable strains in three samples of vaginal fluid. Regardless of the biological material used in PCR to detect carrier animals and the season, the highest MAT sensitivity values were obtained with cut-off 50 compared to 100. The results obtained indicate that, even in the adverse environmental conditions of the semiarid region, leptospires may survive and propagate by alternative routes of transmission, such as sexual, and the presence of PCR-positive genital tracts in ewes suggests that sexual transmission may play an important role in the epidemiology of the disease in sheep in Brazilian semiarid. In addition, it is suggested the use of titer 50 as cut-off point at serology in semiarid conditions.

Influence of breed on the clinical and hemato-biochemical parameters in sheep experimentally infected with Leptospira sp.

Early diagnosis of leptospirosis may aid in a favorable prognosis in infected animals, but there are few reports of clinical and hematochemical changes in the ovine species, nor whether the breed exerts any influence on the response to infection. Thus, the objective of the present study was to evaluate the clinical, biochemical and hematological alterations in Santa Inês and crossbred ewes challenged with Leptospira interrogans serogroup Pomona serovar Kennewicki. Twenty-four sheep were used in this experiment, 12 crossbred (group A) and 12 Santa Inês (group B). In each group, sheep were conjunctivally and intraperitoneally challenged. During 60 days post-infection the ewes were evaluated for the presentation of clinical signs and the blood was collected for hemogram and serum biochemistry. Concentration of urea and creatinine; serum aspartate aminotransferase activity (AST); gamma-glutamyltransferase (GGT); total protein and albumin; total bilirubin, direct and indirect were analyzed. The urine of these sheep was collected for urinalysis. Only two Santa Inês sheep showed blood in the urine. Clinical signs implicated in Leptospira sp. infection were not identified. Some sheep had anemia, especially crossbred. However, anemia may be attributed to the more effective cellular response that has been identified in crossbred animals. Only one animal presented leukocytosis with neutrophilia, while 11 presented atypical leukopenia, especially those of the Santa Inês breed. There was a decrease in total protein and albumin, as well as the increase in gammaglutamyltranspeptidase (GGT), especially in Santa Inês sheep. The level of aspartate aminotransferase (AST) remained within the normal range for the species. A significant increase (p < 0.05) in conjugated bilirubin levels in challenged animals was detected. Only one sheep intraperitoneally challenged presented a high level of urea in the blood, but the creatinine level remained within the normal range. The intraperitoneal route was responsible for more significant changes (p < 0.05) in the hemogram and biochemistry when compared to the conjunctival route. The results indicate that crossbred sheep have a more efficient cellular response than Santa Inês sheep, which may confer a greater resistance to infection. Clinical signs are not good parameters to follow the development of leptospirosis in crossbred and Santa Inês breed. Hematological and biochemical analyzes were useful in the detection of anemia and possible liver changes caused by leptospirosis. The intraperitoneal route was able to cause more conclusive alterations of the analyzed parameters, however, it is possible that the alterations caused by the conjunctival route reproduce in a more faithful way what happens in a natural situation of infection.