Extra-renal bovine leptospirosis: Molecular characterization of the Leptospira interrogans Sejroe serogroup on the uterus of non-pregnant cows.

Bovine genital leptospirosis is a chronic disease that causes reproductive disorders such as abortions, stillbirths, and estrus repetition, as well as economic losses. Despite clinical signs related to reproductive failure, the majority of studies have focused on the detection of Leptospira spp. in the urine, while few have considered the reproductive tract. Consequently, the aim of the present study was to investigate the uterus as an important extra-renal site of leptospiral infection in cows. A total of 42 non-pregnant cows were studied at a slaughterhouse. Blood samples and uterine fragments were collected for serology and molecular analysis, respectively. Concerning serologic results, 20.5 % presented as reactive, all of them against the Sejroe serogroup. Regarding lipL32 PCR, 26.2 % (11/42) of samples were positive for pathogenic Leptospira sp. Sequencing the secY gene short region enabled nine strains to be characterized, all of which were L. interrogans, with high identity (98.8 %-99.8 %) with serovar Hardjo. The use of molecular tools substantially improved the sensitivity of Leptospira sp. detection at species level and demonstrated that the uterus is an important site of bovine leptospiral infection. The findings of the present study reinforce our understanding that leptospiral uterine infection are associated to members of the Sejroe serogroup.

A systematic review of leptospirosis on wild animals in Latin America.

Leptospirosis is a bacterial systemic infection which affects domestic animals and wildlife, as well as humans. Many wild animals act as reservoirs of leptospires. Nevertheless, the real role of wildlife animals as source of infection to livestock and humans, as well as the most important reservoirs and leptospiral strains remains unclear. This systematic review assesses the available data about wildlife and their biomes in Latin America, concerning to leptospiral infection. In addition, we discuss the development of the research on leptospirosis in wildlife in this region. After the application of exclusion criteria, 79 papers were analyzed, comprising 186 species, 122 genus, 53 families, and 19 orders from four classes. Mammals were the most studied class, followed by Amphibian, Reptile, and Aves. The Icterohaemorrhagiae serogroup was predominant in most biomes and many orders. A small number of antigens detected the majority of seroreactive animals of each class, and a smaller panel may be used at microscopic agglutination test. Further studies must always consider edaphoclimatic conditions besides only host class or species, in order to obtain a broader understanding of the wild epidemiological cycle of leptospirosis in the region. In conclusion, direct and indirect evidences demonstrate that leptospirosis is largely widespread among wildlife in all biomes of Latin America. Moreover, more research on the role of wildlife on the epidemiology of leptospirosis and its impact on livestock and public health are required, particularly focusing on direct detection of the agent.

Plurality of Leptospira strains on slaughtered animals suggest a broader concept of adaptability of leptospires to cattle.

Leptospirosis in bovines is in majority determined by the host-adapted serovars, mainly Hardjo (types Hardjoprajitno and Hardjobovis), that belong to the serogroup Sejroe. Members of other serogroups as Pomona and Tarassovi have been eventually reported, mainly when outbreaks occurs. Nevertheless, the real role of other strains (non-Hardjo) on determining disease or being transmitted by cattle free of apparent clinical signs of acute infection remains to be elucidated. In that context, the aim of the present study was to investigate the hypothesis that strains of serovars/serogroups other than Hardjo may also be maintained and shed by cattle free of clinical signs. Samples of urine and/or vaginal fluid were collected from 697 bovines from a slaughterhouse located close to Rio de Janeiro, Brazil. Culturing yielded 19 isolates what represents the largest number ever obtained in Brazil on similar studies. These strains were serogrouped and genetically characterized. Fifteen of those were described in other papers and four are first described on the present study. Isolates belong to three different species (Leptospira santarosai, L. alstonii and L. interrogans) and five serogroups (Sarmin, Tarassovi, Shermani, Grippotyphosa and Sejroe). The majority (84.2%) of the isolates belongs to the species L. santarosai, the most prevalent species on cattle in the studied region. Non-Hardjo (non-Sejroe) strains represent 57.9% of the isolates, what indicates an unexpected high diversity of serogroups obtained from these cattle. This suggest that non-Hardjo (non-Sejroe) strains may also be maintained and shed by cattle and that finding must be considered in the epidemiology and control of the disease.

Seronegativity of bovines face to their own recovered leptospiral isolates.

Leptospirosis is an important cause of reproductive failure in cattle. The standard diagnostic tool (MAT) is recommended for herd but not for individual diagnosis. The aim of this study was to evaluate the humoral response of bovines face to their own recovered isolates. A total of 25 bovine from which leptospires were recovered were tested by MAT against reference strains and their own isolates. Only three cows (12%) presented seroreactivity against their own isolates. This study demonstrates that cattle may not react against their own isolates and highlights the importance of interpreting serological negative results with caution.

A systematic review of leptospirosis on dogs, pigs, and horses in Latin America.

Leptospirosis is a worldwide zoonosis which can affect many species. Control programs need accurate diagnosis to be successful, and currently, diagnosis relies on serology. It presents three main issues: the sampling, the antigen panel, and the cutoff point. Herein, we propose a systematic review on leptospirosis among dogs, pigs, and horses in Latin America in order to improve the understanding of the seroepidemiology of leptospirosis in these species in the region as well as the temporal development of the research on this topic and, consequently, improve the chances of success on control programs. Internet databases were consulted over 2015. Inclusion criteria included serosurvey using MAT; a relevant number of animals; the presence in the antigen panel of at least one representative of serogroups Icterohaemorrhagiae and Canicola for dogs, Icterohaemorrhagiae, Australis, and Pomona for pigs, and Icterohaemorrhagiae and Australis for horses; and a cutoff point of ≥100. Overall, 240 papers were studied, of which 87 referred to dogs, 66 to pigs, 39 to horses, and 48 to more than one of the studied species. In relation to those that met all the inclusion criteria, it was 45 (66.2%) in dogs, 23 (41.8%) in pigs, and 23 (63.9%) in horses. Leptospirosis is widespread in Latin America. Predominant serogroups are Canicola to dogs and Icterohaemorrhagiae to pigs and horses. Therefore, research on animal leptospirosis should be encouraged in Latin America, in order to reach a greater standardization in studies and then achieve better results on control programs.

Usage of Leptospira spp. local strains as antigens increases the sensitivity of the serodiagnosis of bovine leptospirosis.

Leptospirosis is a zoonotic disease that occurs worldwide, particularly in tropical countries. In livestock the agent is responsible for reproductive problems such as infertility and abortion. Serogroup Sejroe, particularly serovar Hardjo, prevails in cattle in several regions. The microscopic agglutination test (MAT) is the current method for diagnosing leptospirosis. It has been proposed that the inclusion of local strains could detect a larger set of seroreactive animals. In that context, the aim of the present study was to evaluate if the usage of local strains as antigens increases the sensitivity of the serodiagnosis of bovine leptospirosis. Blood and urine samples were collected from 314 bovines from several herds randomly selected in a slaughterhouse in Rio de Janeiro, Brazil. Serological diagnosis was made with MAT using a 21 reference-strains panel (MAT21). Additionally, 12 local strains (MAT33) were included as antigens. PCR was performed with the urine samples and it was positive on 71 out of 222 samples (31.9%). MAT21 identified as seroreactive 173 (55.1%) out of the 314 animals studied, with Sejroe the most common (38.1%). In MAT33, 204 (65.0%) animals were seroreactive with a significant increase on seroreactivity (9.9%). In conclusion, MAT presented with a significant increase of sensitivity when local strains were used as antigens. Among the local strains, 2013_U152 (KP263062) (serogroup Shermani) and 2013_U280 (KP263069) (serogroup Grippotyphosa) showed to be more antigenic.