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The aim of this systematic review was to identify articles on prevalence of leptospirosis in stray and sheltered dogs worldwide and access the methodological quality of the recovered papers. Six databases (CABI, Cochrane, Pubmed, Scielo, Scopus and Web of Science) were searched, without restriction on year or location where the studies were performed. The search recovered 476 articles and 60 were selected for analysis according to quality criteria. None of the selected articles showed a complete explanation for the sample size adopted (probabilistic sampling), leading to the impossibility of recalculation of leptospirosis prevalence for stray or sheltered dogs. Among the analyzed papers 43.33% (26/60) showed five of the ten quality criteria analyzed, 16.67% (10/60) three, 15.00% (9/60) four, 10.00% (6/60) six, 6.67% (4/60) eight, only 5.00% (3/60) showed nine of the ten criteria analyzed, whereas two papers showed two [1.67% (1/60)] and seven [1.67% (1/60)] of the ten criteria assessed. The majority of the papers were published in the Americas [45.00% (27/60)] and in the last sixteen years (2003 to 2019) [81.67% (49/60)], and most of the sampled dogs were stray dogs [65.00% (39/60)]. The leptospirosis diagnostic test used more frequently was Micro Agglutination Test (MAT) [78.33% (47/60)] followed by polymerase Chain Reaction (PCR) [23.33% (14/60)], whereas the most common serovars identified were Canicola [71.43% (35/49)], Icterohaemohrragiae [65.31% (32/49)], Grippotyphosa [40.82% (20/49)] and Pomona [40.82% (20/49)]. In conclusion, our results showed that Leptospira spp. is present in stray and sheltered dogs worldwide, but the complete comprehension of the prevalence of leptospirosis in these populations could not be achieved due to the low methodologic quality of the recovered studies about leptospirosis in stray and sheltered dogs.

Changes were made to the original formulation of the EMJH medium (Ellinghausen-McCullough-Johnson-Harris) enrichment and some aspects such as growth time of Leptospira and utilization in the microscopic agglutination test (MAT) were evaluated and compared to the original enrichment and to a commercially available enrichment (DIFCOÔ). Leptospira samples (24 antigens) that make up our panel of antigens used in MAT were used, among them, reference and autochthonous strains isolated in Brazil. The samples were grown individually in the EMJH medium under the three previously mentioned conditions (adapted enrichment, original enrichment and commercial enrichment). In addition, 89 blood serums from domestic and wild animals were analyzed by MAT using the antigens grown in these media. All samples tested grew efficiently with the adapted enrichment, and the MAT results were satisfactory. Therefore, other laboratories could also benefit from the use of this adapted enrichment when culturing the Leptospira strains applied in their MAT panels. Resumen Realizamos cambios en la formulación original del enriquecimiento del medio Ellinghausen-McCullough-Johnson-Harris y evaluamos algunos aspectos, como el tiempo de crecimiento de Leptospira y la utilización en la prueba de aglutinación microscópica (MAT), comparándolos con el enriquecimiento original y un medio de enriquecimiento comercialmente disponible (DIFCOÔ). Se usaron muestras de Leptospira (24 antígenos) que componen nuestro panel de antígenos utilizados en la MAT, entre ellos, cepas autóctonas y de referencia aisladas en Brasil. Las muestras se cultivaron individualmente en medio Ellinghausen-McCullough-Johnson-Harris en las tres condiciones mencionadas (enriquecimiento adaptado, enriquecimiento original y enriquecimiento comercial). Adicionalmente, 89 sueros de sangre de animales domésticos y salvajes fueron analizados por MAT usando los antígenos cultivados en estos medios. Todas las muestras analizadas crecieron eficientemente con el enriquecimiento adaptado y los resultados de la MAT fueron satisfactorios. Por lo tanto, otros laboratorios también podrían beneficiarse del uso de este enriquecimiento adaptado al cultivar las cepas de Leptospira aplicadas en sus paneles MAT.

Leptospirosis is an emerging infectious disease caused by pathogenic Leptospira spp. A universal vaccine against leptospirosis is likely to require highly conserved epitopes from pathogenic leptospires that are exposed on the bacterial surface and that generate a protective and sterilizing immune response. Our group recently identified several genes predicted to encode TonB-dependent receptors (TBDR) in Leptospira interrogans using a reverse vaccinology approach. Three leptospiral TBDRs were previously described and partially characterized as ferric-citrate, hemin, and cobalamin transporters. In the current study, we designed a fusion protein composed of predicted surface-exposed epitopes from three conserved leptospiral TBDRs. Based on their three-dimensional structural models and the prediction of immunogenic regions, nine putative surface-exposed fragments were selected to compose a recombinant chimeric protein. A Mycobacterium bovis BCG strain expressing this chimeric antigen encoded in the pUP500/PpAN mycobacterial expression vector was used to immunize Syrian hamsters. All animals (20/20) vaccinated with recombinant BCG survived infection with an endpoint dose of L. interrogans (p

Leptospira interrogans is one of the causative agents of human leptospirosis, a zoonotic disease with worldwide distribution. Nowadays, this zoonosis is considered one of the biggest in terms of morbidity and mortality (even considering Dengue, the major arbovirosis affecting humans), having in Brazil 3,800 human cases per year. Currently, difficulties imposed by the absence of a rapid, sensitive diagnostic test that can be used as a routine test for the detection of leptospirosis lead to misdiagnosis and underreported cases. The gold standard diagnostic test for leptospirosis is the microscopic agglutination test (MAT), which presents difficulties in execution and interpretation. Therefore, this review proposes a general view of the epidemiologic situation of the disease in Brazil, in addition to the current contributions in the literature for the development of new diagnostic methods. Amongst them, the gene sequences polymorphism analysis, which presents potential for phylogenetic and populational analysis and genotyping of Leptospira spp. Leptospira interrogans é uma das bactérias causadoras da leptospirose, uma zoonose de ampla distribuição mundial. Atualmente, essa zoonose é considerada uma das que apresenta maiores taxas de morbidade e mortalidade no Brasil (mesmo considerando a Dengue, a maior arbovirose em humanos), com cerca de 3.800 casos humanos por ano documentados. Porém, devido às dificuldades impostas pela ausência de um ensaio de diagnóstico rápido, sensível e que possa ser empregado como teste de rotina para a detecção da leptospirose, essa doença é comumente subnotificada e diagnosticada erroneamente. O teste diagnóstico padrão ouro para a leptospirose é a aglutinação microscópica, o qual apresenta dificuldade de execução e interpretação. Dessa forma, propomos nesta revisão uma visão geral da situação epidemiológica da doença no Brasil, além das contribuições presentes na literatura para o desenvolvimento de novas abordagens diagnósticas. Dentre elas, a análise de polimorfismos de sequências gênicas a qual apresenta potencial para análises filogenéticas, populacionais e de genotipagem de Leptospira spp.

Leptospirosis is a globally prevalent zoonotic disease, and is caused by pathogenic spirochetes from the genus Leptospira. LipL21 and LipL41 are lipoproteins expressed strongly on the outer membrane of pathogenic Leptospira spp. Many studies have shown that both proteins are interesting targets for vaccines and diagnosis. However, their role in host–pathogen interactions remains underexplored. Therefore, we evaluated the capacity of LipL21 and LipL41 to bind with glycosaminoglycans (GAGs), the cell receptors and extracellular matrix, and plasma components by ELISA. Both proteins interacted with collagen IV, laminin, E-cadherin, and elastin dose-dependently. A broad-spectrum binding to plasma components was also observed. Only LipL21 interacted with all the GAG components tested, whereas LipL41 presented a concentration-dependent binding only for chondroitin 4 sulfate. Although, both proteins have the ability to interact with fibrinogen, only LipL21 inhibited fibrin clot formation partially. Both proteins exhibited a decrease in plasminogen binding in the presence of amino caproic acid (ACA), a competitive inhibitor of lysine residues, suggesting that their binding occurs via the kringle domains of plasminogen. LipL41, but not LipL21, was able to convert plasminogen to plasmin, and recruit plasminogen from normal human serum, suggesting that the interaction of this protein with plasminogen may occur in physiological conditions. This work provides the first report demonstrating the capacity of LipL21 and LipL41 to interact with a broad range of host components, highlighting their importance in host–Leptospira interactions.

Leptospirosis is known to determine reproductive disorders on livestock, and Leptospira interrogans and Leptospira borgpetersenii are the most frequently reported species. Leptospira noguchii is an emerging pathogen, but its association with reproductive disease is unclear. We have detected L. noguchii as the agent of an outbreak with reproductive disorders in a Brazilian dairy goat flock. In the kidding season, five out of 10 Saanen had abortions in the final month of pregnancy and two newborn kids had acute clinical signs. After necropsy of three foetuses and one newborn kid, fragments of liver, lung and kidney were submitted to lipL32-PCR. It yielded positive results in at least one fragment from each animal. After, a nested secY-PCR, followed by sequencing, could identify L. noguchii, with 99-100% of identity with sequences obtained from cattle in the same region. For the first time, L. noguchii was detected in goats and, most importantly, the association of this leptospiral species with reproductive failures in ruminants has been demonstrated.

Background The pathogenic Leptospira can survive and contaminate surface water based on physicochemical factors. This study aimed to determine how the physicochemical properties of water sources influence the growth and effect of iron on the gene expression of Leptospira spp. P47. Methods Surface water samples (n=55) were collected and used for Leptospira spp. P47 cultivation. Physicochemical factors, including iron, calcium, magnesium and pH, were analyzed. The association between Leptospira spp. P47 viability at days 5, 10 and 15 with the physicochemical factors were analyzed. In addition, this bacterium was cultured in six selected water samples. The effect of iron in water on HemO and LipL32 gene expression was determined by relative quantification real-time PCR. Results Leptospira viability at day 5 was not significantly correlated with physicochemical factors, while Leptospira viability at day 10 was associated with both pH and iron. The Leptospira viability rate at day 15 had a significantly positive association with pH and iron and a negative association with calcium. HemO expression was significantly increased, mostly in selected water samples and under iron-depleted conditions. Conversely, LipL32 expression was significantly decreased in all water samples. Conclusions Physicochemical factors in natural surface waters are key factors for bacterial survival in the environment, which may increase the chance of Leptospira infection in humans.

La importancia de las cabras en México radica en que su producción se concentra en zonas áridas y semiáridas del norte y el centro del país, y particularmente en la región de la Mixteca en los estados de Oaxaca, Guerrero y Puebla, donde prevalece la pobreza y donde la especie, es capaz de sobrevivir y producir leche y cabritos. El estudio de las enfermedades de las cabras en México es escaso; el INIFAP se ha caracterizado por nunca haber dejado de lado la investigación en esta especie. En esta revisión, se abordan las principales enfermedades que afectan a las cabras en México y se describen las investigaciones que sobre ellas ha desarrollado el INIFAP. Los problemas de salud que enfrentan la mayoría de los rebaños caprinos en los que se ha realizado investigación, demuestran que algunos trastornos reproductivos son causados por enfermedades como brucelosis, leptospirosis y clamidiasis, todas éstas consideradas endémicas y con potencial zoonótico. Por otra parte, los problemas de tipo respiratorio y digestivo son los principales padecimientos que afectan a los cabritos. En las cabras en etapa productiva del territorio nacional, se ha demostrado una frecuencia elevada de animales con artritis encefalitis, enfermedad causada por la infección de Lentivirus de los pequeños rumiantes, paratuberculosis y linfadenitis caseosa, estas últimas de origen bacteriano; una particularidad de estas tres enfermedades es su característica de cronicidad, lo que hace pensar de manera equívoca al productor que su presencia no tiene mayor impacto dentro de los rebaños. Finalmente, se ha logrado detectar la presencia de la fiebre Q, una enfermedad zoonotica actualmente considerada exótica en México, y que en otros países es frecuentemente asociada con trastornos reproductivos, abortos y ocasionalmente, problemas respiratorios en los animales. En las aportaciones del INIFAP hay que destacar el diagnóstico y el control de la brucelosis, que es la principal zoonosis bacteriana en nuestro país; además de haber contribuido a que enfermedades consideradas exóticas en México, se determine que ya son endémicas, y se cuente con las herramientas para su diagnóstico, siendo un reto para nuestro instituto realizar la transferencia de estas tecnologías a los laboratorios de diagnóstico de nuestro país.

Background Refugees in Malaysia, who are afflicted by poverty, conflict and poor health, are vulnerable to a range of zoonotic infections in the deprived environmental and social conditions under which they live. Exposure to infections such as leptospirosis, for which rodents are primary hosts, is of particular concern. Methods A wellness program was conducted to determine the presence of antibodies against Leptospira (seroprevalence) in 11 refugee community schools and centers in the Klang Valley, Malaysia. A total of 433 samples were assessed for IgG and IgM antibodies against Leptospira, using enzyme-linked immunosorbent assays (ELISA). Results Overall Leptospira seroprevalence was 24.7%, with 3.0% being seropositive for anti-Leptospira IgG and 21.7% for anti-Leptospira IgM. Factors significantly associated with overall Leptospira seroprevalence included: age, ethnicity, pet ownership, knowledge of disease and awareness of disease fatality. For IgM seroprevalence, significant risk factors included sex, ethnicity, eating habits with hands, pet ownership, the presence of rats, walking in bare feet and water recreation visits. Conclusions These findings highlight the need for improvements in health and well-being among the refugee community through disease awareness programs and provision of healthy behavior programs, particularly in hygiene and sanitation through community engagement activities.

Background Leptospirosis diagnoses have increased in Sarawak, Malaysia in recent years. Methods To better understand the burden of disease and associated risk factors, we evaluated 147 patients presenting with clinical leptospirosis to local hospitals in Sarawak, Malaysia for the presence of Leptospira and associated antibodies. Sera and urine specimens collected during the acute illness phase were assessed via a commercially available rapid diagnostic test (Leptorapide, Linnodee Ltd., Antrim, Northern Ireland), an ELISA IgM assay (Leptospira IgM ELISA, PanBio, Queensland, Australia) and a pan-Leptospira real-time PCR (qPCR) assay to estimate disease prevalence and diagnostic accuracy of each method. Microagglutination testing was performed on a subset of samples. Results Overall, 45 out of 147 patients (30.6%) showed evidence of leptospires through qPCR in either one or both sera (20 patients) or urine (33 patients), and an additional ten (6.8%) were considered positive through serological testing, for an overall prevalence of 37.4% within the study population. However, each diagnostic method individually yielded disparate prevalence estimates: rapid test 42.2% for sera and 30.5% for urine, ELISA 15.0% for sera, qPCR 13.8% for sera and 23.4% for urine. Molecular characterization of a subset of positive samples by conventional PCR identified the bacterial species as Leptospira interrogans in 4 specimens. A multivariate risk factor analysis for the outcome of leptospirosis identified having completed primary school (OR = 2.5; 95 CI% 1.0–6.4) and weekly clothes-washing in local rivers (OR = 10.6; 95 CI% 1.4–214.8) with increased likelihood of leptospirosis when compared with those who had not. Conclusion Overall, the data suggest a relatively high prevalence of leptospirosis in the study population. The low sensitivities of the rapid diagnostic test and ELISA assay against qPCR highlight a need for better screening tools.

Background. In presence the epizootiological situation on leptospirosis in the territories adjacent to Lake Baikal is not studied well. The showing up of natural focus, discovering of host animal’s species composition and etiological structure of leptospirosis causative agents are essential to plan any epidemiological response and to organize the specific prophylaxis.Aim: to analyze contemporary characteristics of leptospirosis natural foci at the territories adjacent to Lake Baikal.Materials and methods. From 2011 till 2020 the territory of nine Irkutsk Region's administrative districts was investigated. Trapping has been done at four kind of sites (wetland, meadow, forest-shrub, steppificated meadow) and boroughs. 1152 small mammals which belonged to 35 species were collected. Collected samples were studied by complex of methods. Serogroup belonging was detected by micro agglutination and decomposition test, and genomic species – by Microflex LT mass analyzer.Results. A leptospirosis natural foci in the Irkutsk city boundary was newly founded and there were isolated (from small mammals) four pathogenic cultures of leptospiras belonging to Javanica serogroup. Main hosts of causative agents at the territories adjacent to Lake Baikal were defined (insect-eating mammals: tundra shrew, largetoothed Siberian shrew, even-toothed shrew, shrew-mouse, and rodents: root vole, ground vole, narrow-skulled vole, sewer rat). By micro agglutination were defined pathogenic leptospirosis serogroups (Icterohaemorrhagiae, Grippotyphosa, Javanica, Pomona, Sejroe, Autumnalis, Australis), all over 2.4 ± 0.45 % seropositive. Antibody dilution at survey animals ranged from 1 : 20 to 1 : 640. On the grounds of PCR results the mean value of infected animals in studied sample was 16.4 ± 1.14 %. Positive PCR findings were at 19 from 35 animal species.Conclusion. At the territories adjacent to Lake Baikal In the Baikal region, the proportion of animals that have had contact with pathogenic leptospira is quite high. Natural focus of this infection are highly pervasive in this region, they are evolved mainly in dewy sites.

International audience; Spirochetes can be distinguished from other bacteria by their spiral-shaped morphology and subpolar periplasmic flagella. This study focused on FlhF and FlhG, which control the spatial and numerical regulation of flagella in many exoflagellated bacteria, in the spirochete Leptospira. In contrast to flhF which seems to be essential in Leptospira, we demonstrated that flhG- mutants in both the saprophyte L. biflexa and the pathogen L. interrogans were less motile than the wild-type strains in gel-like environments but not hyperflagellated as reported previously in other bacteria. Cryo-electron tomography revealed that the distance between the flagellar basal body and the tip of the cell decreased significantly in the flhG- mutant in comparison to wild-type and complemented strains. Additionally, comparative transcriptome analyses of L. biflexa flhG- and wild-type strains showed that FlhG acts as a negative regulator of transcription of some flagellar genes. We found that the L. interrogans flhG- mutant was attenuated for virulence in the hamster model. Cross-species complementation also showed that flhG is not interchangeable between species. Our results indicate that FlhF and FlhG in Leptospira contribute to governing cell motility but our data support the hypothesis that FlhF and FlhG function differently in each bacterial species, including among spirochetes.

Neuroleptospirosis is a rare disease caused by pathogenic Leptospira interrogans in humans; however, it has not been fully studied in animals. A young wild raccoon dog was found convulsing in the recumbent position and died the next day. Histologic examination revealed nonsuppurative meningoencephalitis in the cerebrum, cerebellum, midbrain, and medulla oblongata. The lesions consisted of mixed infiltrates of Iba1-positive macrophages and CD3-positive T cells, with a small number of CD79α-positive B cells and myeloperoxidase-positive neutrophils. In the frontal cortex, perivascular cuffs and adjacent microglial nodules were distributed diffusely, especially in the molecular layer. Glial nodules were comprised of Iba1- and myeloperoxidase-positive activated microglia. Immunohistochemistry revealed leptospires in mononuclear cell perivascular cuffs, but not in glial nodules. Neuroleptospirosis was accompanied by Leptospira-related nonsuppurative interstitial nephritis, pulmonary edema and hemorrhage, and coronary periarteritis, as well as Toxocara tanuki in the small intestine and nonspecific foreign-body granulomas in the lungs and stomach.

Leptospira interrogans spirochetes belong to a group of pathogens of particularly dangerous infections that cause leptospirosis in many species of wild, farm and domestic animals, as well as humans. Significant economic damage caused by this disease to livestock around the world, numerous cases of death from leptospirosis, and mainly waterborne transmission of the infection determine the topicality of studying all the aspects of the existence of L. interrogans in freshwater ecosystems. The objective of our study was to determine the nature of ecological relationships between L. interrogans and green algae. We used green algae of the Desmodesmus brasiliensis species as a model object. In the experiments, sterile algae culture filtrates were added to L. interrogans containers grown at 27–28 °C. Comparison of the content of spirochete cells in the experiment and control samples, conducted 24 hours after exposure, showed that the reproduction of L. interrogans is markedly inhibited in the samples containing algae secretions at the dilutions of 1:10 and 1:100. In order to reduce resistance to the allelopathic influence of D. brasiliensis algae, leptospira strains were located as follows: Pomona, Australis, Hebdomadis, Canicola, Sejroe, Icterohaemorrhagiae, Grippotyphosa, Tarassovi. The obtained results indicate a complex intraspecific structure of L. interrogans and their high ecological plasticity. In addition, experimental data indicate that, due to the release of biologically active substances into the aquatic environment, green algae have the potential to affect the dynamics of a number of leptospirosis pathogens in situ. Given the important epidemiological and epizootic significance of pathogenic leptospira, further research is necessary to determine the patterns of their interactions and existence in the environment.

Leptospirosis is a zoonotic neglected disease of worldwide public health concern. Leptospira species can infect a wide range of wild and domestic mammals and lead to a spectrum of disease, including severe and fatal forms. Herein, we report for the first time a fatal Leptospira interrogans infection in a free-ranging nonhuman primate (NHP), a black-tufted marmoset. Icterus, pulmonary haemorrhage, interstitial nephritis, and hepatocellular dissociation were the main findings raising the suspicion of leptospirosis. Diagnostic confirmation was based on specific immunohistochemical and PCR assays for Leptospira species. Immunolocalization of leptospiral antigens and identification of pathogenic species (L. interrogans species) were important for better understanding the pathogenesis of the disease. One Health-related implications of free-ranging NHPs in anthropized areas and transmission dynamics of human and animal leptospirosis are discussed.

Made available in DSpace on 2022-05-01T08:44:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 The present study investigated the serum microscopic agglutination test (MAT) among 203 bovine bulls with reproduction by natural means, without apparent signs of orchitis or inflammation of accessory reproductive glands. Simultaneously, the semen of all bulls was subjected to sperm viability analysis and PCR based on the 16S rRNA gene. PCR-positive results of semen samples were confirmed by sequencing. A modified seminal plasma agglutination (MSPA) test, replacing the blood serum of all bulls in the MAT with seminal plasma was performed as well. Eight (8/203 = 3.9%) semen samples from bulls were considered nonviable (necrospermia and azoospermia) without relation to the PCR diagnosis. No agglutinin titers were identified in MSPA test. A high frequency (132/203 = 65%) of leptospiral agglutinin titers was identified in the MAT, particularly for the Sejroe serogroup (Hardjo CTG, 100/203 = 49.3%; Wolffi 74/203 = 36.4%; Guaricura 72/203 = 35.5%; and Hardjoprajitno 56/203 = 27.6%). Three (3/203 = 1.5%) semen samples of bulls were positive in the PCR, but these results were not confirmed by sequencing. The high frequency of serovars from the Sejroe serogroup typically adapted to bovines indicates the need for measures for the prophylaxis/control of the pathogen on the sampled farms. Discrepancies among the MAT, sperm viability, and molecular detection of leptospires in semen highlight the need for a combination of methods to diagnose leptospirosis in bovine bulls. To our knowledge, modified seminal plasma agglutination is described for the first time here to investigate anti-Leptospira antibodies produced locally in the genital tract in the diagnosis of bovine leptospirosis among bulls that reproduce by natural means. Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine an Animal Science São Paulo State University - UNESP Santo Amaro University - UNISA Faculty of Veterinary Medicine University of Oeste Paulista Technology Faculty - FATEC APTA Beef Cattle Center Institute of Animal Science University of São Paulo - USP Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine an Animal Science São Paulo State University - UNESP

Background and Aim: For understanding the epidemiology of leptospirosis, the confined abundance of several species of pathogenic leptospires and knowledge on the serovar(s) prevalent in the reservoir and carrier hosts may be a useful indicator of transmission to incidental/accidental hosts in a geographical niche. The present study was carried out to ascertain the frequency distribution of Leptospira serovars and the prevalence of anti-leptospiral antibodies in small ruminants (sheep and goats) in the epidemiological units (villages) in the coastal districts of enzootic regions in South Peninsular India. Materials and Methods: A total of 1167 serum samples (sheep n=299 and goats n=868) from apparently healthy animals, randomly collected from various epidemiological units were tested in microscopic agglutination test (MAT) using 18 reference Leptospira serovars antigens. Results: The overall seroprevalence of 40% (at 95% confidence intervals [CI]: 36.82-42.43) in small ruminants (44% [95% CI: 40.49-52.26] in sheep and 38% [95% CI: 34.96-41.41] in goats) was observed with the predominance of Icterohaemorrhagiae, Javanica, Australis, Hurstbridge, and Pyrogenes serogroup anti-leptospiral antibodies in the studied region. The Chi-squared test revealed that the presence of anti-leptospiral antibodies is significantly not independent (associated) across the administrative division (Chi-square=105.80, p

Leptospirosis causes abortion, premature birth, and stillbirth in cattle, but the mechanisms remain unclear. Infected cattle shed Leptospira intermittently and present a range of clinical symptoms, making diagnosis difficult. The primary route of Leptospira transmission in any animal is the colonization of the renal tubule and excretion by urine; however, Leptospira can also colonize the female reproductive tract of cows and can be transmitted by semen. Vaccination against Leptospira in the US is routine in cattle, but immunity is not guaranteed. The cell wall of Leptospira contains toll-like receptor agonists including peptidoglycan and lipopolysaccharide. The capacity of Leptospira to initiate an innate inflammatory response from uterine endometrial cells is unknown but may be a cause of reproductive failure. Using cell culture, we tested the capacity of bovine endometrial epithelial cells or human monocytes to elicit an inflammatory response to Leptospira borgpetersenii serovar Hardjo strain TC273. Cells were exposed to either heat-killed Leptospira, Leptospira outer membrane, Escherichia coli lipopolysaccharide, Pam3CSK4 or medium alone for 2 to 24 h. Exposure of bovine endometrial epithelial cells or human monocytes to heat-killed Leptospira or Leptospira outer membrane did not induce the expression of IL1A, IL1B, IL6, or CXCL8, while exposure to E. coli lipopolysaccharide or Pam3CSK4 increased the expression of IL1A, IL1B, IL6, and CXCL8 compared to control cells. This data suggest that Leptospira does not trigger a classical inflammatory response in endometrial cells. Understanding the interaction between Leptospira and the female reproductive tract is important in determining the mechanisms of Leptospirosis associated with reproductive failure. Lay summary Cows infected with the Leptospira have abortion and stillbirth. It is not known how Leptospira causes pregnancy failure in the cow. We tested if Leptospira causes inflammation in cells of the uterus which triggers pregnancy failure. We collected cells from the uterus of healthy cows at the abattoir and placed them into culture with Leptospira and measured the expression of genes associated with inflammation. To our surprise, cells of the uterus did not respond to Leptospira; however, the same cells did respond to other disease-causing bacteria found in the uterus. This suggests that cells of the uterus can recognize bacteria and produce an inflammatory reaction but not in response to Leptospira. This finding suggests the immune system of the uterus cannot detect Leptospira which may go on to cause reproductive failure in cows. Understanding how Leptospira interact with cells of the uterus will help reduce pregnancy failure of cows with leptospirosis.

Rodents are the most abundant and diversified order of living mammals in the world. Since the middle ages they are known to contribute to human diseases, eg. black rats associated with distribution of plague. In modern times also rodents form a threat for public health and a number of pathogens are directly or indirectly transmitted by rodents. Of 2277 extant rodent species, 217 species are reservoirs harboring 66 zoonoses caused by viruses, bacteria, fungi, helminths, and protozoa and play a major role in their transmission and spread in different ways. Among the most important diseases in terms of public health are salmonellosis, plague, leptospirosis, leishmaniasis, toxoplasmosis, rat-bit fever, taeniasis-like Capillaria hepatica, zoonotic babesiosis, Lassa fever, hemorrhagic fever with renal syndrome (HFRS), and the hantavirus cardiopulmonary syndrome (HCPS). In addition, Arenaviruses are responsible for South American Hemorrhagic Fevers (SAHF). Rodents may harbor different complex bacteria, such as Mycobacterium tuberculosis and Mycobacterium microti, Escherichia coli, agents of Tularemia, Tick-borne relapsing fever, Bartonellosis, Listeriosis, Lyme disease, Q fever, Ehrlichiosis and others. Recent changes in the global climate and ecosystem composition led to the spread of rodents and rodent-borne pathogens globally. Rodent control in rural environments relies mainly on the use of rodenticides. Other management methods which can be applied are trapping, habitat management (e.g., proofing, sanitation), and biocontrol (e.g., wild or domestic predators, rodent pathogens).

Zoonoses are primarily recognized as animal diseases that are transmitted to human beings through various routes. These diseases have a serious impact on public health as well as economy throughout the world. Leptospirosis is an emerging and re-emerging infectious zoonotic disease with global public health implications in terms of morbidity and mortality of humans and livestock. The disease can occur in sporadic as well as in epidemic forms. The source of infection is exogenous. Leptospirosis is a life threatening disease that causes 10.3 million cases and 58900 deaths each year worldwide. Rodents are considered the main reservoir of Leptospira. The contact of the skin with moist soil, water, and vegetation contaminated with urine of the infected animals and penetration of the organism to the skin, mucous membrane through abrasion, cut, and injury help in the transmission of the infection. Leptospirosis presents most important public health crisis that exists at the animals-humans-ecosystem interface. A strategic break in the relationship between the host-pathogen and their ecosystem could provide effective control of several possible zoonoses including leptospirosis. Global strategy to broaden interdisciplinary partnerships and coordination in all areas of health care for humans, livestock and the community is required.

The aim of the work was to rank statistical models for assessing the contribution of a number of factors that determine the epizootic situation on natural-focal infectious diseases in the communities of non-synanthropic small mammals (SM) in the forests of Ekaterinburg.Materials and methods. The SM survey was carried out for three years in the summer-autumn period. The animals were caught with snap tpaps on a standard bread bait, 9705 trap-nights were worked out. To identify infections (hemorrhagic fever with renal syndrome, tularemia, leptospirosis, yersiniosis, pseudotuberculosis), 333 SM specimens (rodents and shrews of six species) were used, randomly taken from the general sample. To determine the pathogens, the methods of enzyme immunoassay and polymerase chain reaction were used. The cumulative infection rate of small mammals with all studied natural-focal infections was evaluated. Statistical data processing was carried out from the standpoint of a multi-model approach. The selection of models was performed using the Akaike information criterion with the calculation of the sum of the weights of the models based on their full spectrum (SW).Results and discussion. Carriers of all of the mentioned above infections, except for pseudotuberculosis, have been identified in the forest parks of Ekaterinburg. According to the ranking, the model with three predictors received the highest weight: species, year, and the abundance of SM of the previous year in a particular habitat. The most significant predictors for the full spectrum of models are the year (SW=1), species (SW=0.6), abundance of animals in the current and previous year (SW=0.48). The distribution of positive samples by species of small mammals corresponded to their ranking by abundance in the community. The influence of the factors “season” (summer or autumn) and “area” (place of capture of animals) turned out to be insignificant on the scale of the studies (SW = 0.3 and 0.16, respectively). The percentage of infected samples in total for all infections varied significantly by location and year (0–60%). The possible significance of unaccounted factors is discussed: landscape features and the mode of using forest parks, dispersal of small mammals. The conclusion is made about the usefulness of the multi-model approach in the analysis of the data from epizootiological studies.

Leptospirosis is an emerging zoonotic disease endemic in Kerala and close monitoring of emerging serovars is essential to adopt appropriate control strategies. Multi-Locus Sequence Typing (MLST) was reported to be less expensive compared to other cumbersome methods like whole genome sequencing. The present study was conducted to obtain isolates of Leptospira from infected animals and rats and for the identification of serovars using MLST. A total of 205 blood samples (dog, cat, cattle, goat), 43 urine samples (dog, cattle) and post-mortem kidney samples from various animals such as dog (n=12), cattle (n=2) and rat (n=25) were collected and subjected to polymerase chain reaction (PCR) using G1/G2 primers to identify the pathogenic Leptospira. Fifteen samples were found to be positive, these samples when inoculated in the Ellinghausen- McCullough-Johnson-Harris (EMJH) semi-solid medium to obtain ten isolates. These ten isolates were further subjected to secY, icdA and GyraseB PCR and sequenced. The obtained sequences were analysed using BLAST and were fed into specified MLST database of Leptospira scheme-3, the allelic profile and sequence type were generated. The MLST results obtained in the study indicated that the isolates S24 and S33 belonged to serovar Canicola, S40 and 47 were Sejroe and S19, S27, S55, S69 and S71 were Bataviae, Autumnalis, Pomona, Icterohaemorraghiae and Australis, respectively. It was concluded that MLST is a convenient method for identifying leptospiral serovars.

Leptospirosis is an endemic disease caused by Leptospira spp., a bacterium that affects animals and humans. In recent years, the number of reports of leptospirosis in wild animals has increased, which highlights the need to study the infectious agents in these animals. In this study, a duplex PCR for the detection of leptospiral DNA was performed on 50 kidney samples from bats, and a MAT (Microscopic Agglutination Test) for serological detection of anti-leptospiral antibodies was applied to 47 serum samples from bats from different regions of Buenos Aires Province, Argentina. DNA was extracted using Chelex-100 and duplex PCR was performed by targeting the detection of genes secY and flaB, of pathogenic Leptospira spp. Of the 50 kidney samples, 3 were positive for Eumops sp. and Tadarida brasiliensis by duplex PCR. Of the 47 serum samples, 12 were positive for different serovars: Leptospira interrogans serovars Icterohaemorrhagiae, Cynopteri and Bataviae, and Leptospira borgpetersenii serovar Ballum. This is the first report of the detection of pathogenic leptospires by serology in bats belonging to the T. brasiliensis and Eptesicus furinalis species in Argentina. In addition, this is the first report of the detection of pathogenic leptospiral DNA by PCR in T. brasiliensis species. The detection of Leptospira spp. in these wild animals shows that they may play an important role as wildlife reservoirs of leptospires.

In 2017 leptospirosis cases occur in new areas in Bantul where cases have never been previously reported. Cases still occur in 2018 with an interval of two months after the first case. Preventive and curative efforts in the context of controlling cases have been carried out by the local health office, in addition to these control efforts, it is also necessary to know the presence of Leptospira in rats and water bodies in the area as well as the history of patients transmission. This study was conducted to provide epidemiological data, especially the status of Leptospira in rats as reservoir animal and water bodies as well as a history of patient activity as basic data for leptospirosis control in Gilangharjo Village. This cross-sectional study was conducted at the case location in RT 03 Jodog, Gilangharjo Village, Pandak District, Bantul Regency, which was conducted in April 2018. Leptospira infection status was carried out on rats and water bodies by molecular examination, while the history of transmission based on interviews with patients and the result of the examination of Leptospira status. Rat species caught were dominated by R.tanezumi, R. norvegicus and B. indica. Positive rat infected with Leptospira were found in B. indica while in water bodies Leptospira contamination was not found. Infected rat can be a source of transmission for humans and other animals. Most of the patient's activities were carried out in the fields and at home, all of the patients had wounds that were not treated, and had a history of contact with rats that might transmit Leptospira to humans.

The spatial distribution of environmental conditions may influence the dynamics of vectorborne diseases like leptospirosis. This study aims to investigate the global and localised relationships between leptospirosis with selected environmental variables. The association between environmental variables and the spatial density of geocoded leptospirosis cases was determined using global Poisson regression (GPR) and geographically weighted Poisson regression (GWPR). A higher prevalence of leptospirosis was detected in areas with higher water vapour pressure (exp(â): 1.12; 95% CI: 1.02 - 1.25) and annual precipitation (exp(â): 1.15; 95% CI: 1.02 - 1.31), with lower precipitation in the driest month (exp(â): 0.85; 95% CI: 0.75 - 0.96) and the wettest quarter (exp(â): 0.88; 95% CI: 0.77 - 1.00). Water vapor pressure (WVP) varied the most in the hotspot regions with a standard deviation of 0.62 (LQ: 0.15; UQ; 0.99) while the least variation was observed in annual precipitation (ANNP) with a standard deviation of 0.14 (LQ: 0.11; UQ; 0.30). The reduction in AICc value from 519.73 to 443.49 indicates that the GWPR model is able to identify the spatially varying correlation between leptospirosis and selected environmental variables. The results of the localised relationships in this study could be used to formulate spatially targeted interventions. This would be particularly useful in localities with a strong environmental or socio-demographical determinants for the transmission of leptospirosis.

Leptospirosis is a serious bacterial disease that affects both humans and animals. A wide range of symptoms have been described in humans; the disease in dogs is commonly associated with kidney and/or liver disease. In Malaysia, information about the common serovars infecting dogs is limited. Therefore, we investigated the occurrences of leptospirosis in 124 pet dogs diagnosed with kidney and/or liver disease. Blood, urine, abdominal effusion, and/or kidney and liver were collected from the dogs. Based on microscopic agglutination testing, 53 of 124 (42.7%) dogs were seropositive for leptospiral exposure. Sera were frequently positive to serovars Bataviae ( n = 12), Javanica ( n = 10), and Icterohaemorrhagiae ( n = 10). Direct detection using PCR showed that 42 of 124 (33.9%) of the whole blood and 36 of 113 (31.9%) urine samples were positive for pathogenic Leptospira spp. By PCR, 2 of 23 (9.1%) kidney and 2 of 23 (9.1%) liver were positive for pathogenic Leptospira spp. Abdominal effusion from 4 dogs were PCR-positive for pathogenic Leptospira spp. The species detected were L. interrogans, L. borgpetersenii, L. kirschneri, and L. kmetyi by partial 16S rRNA sequencing. We further identified and characterized 11 Leptospira spp. isolates from 8 dogs as serovars Bataviae, Javanica, and Australis. The mortality rate of the Leptospira-infected dogs was high (18 of 53; 34%).

BackgroundLeptospirosis is an endemic zoonosis in Brazil, with a great impact on human and animal health. Although dogs are frequently infected by pathogenicLeptospira, the current epidemiological understanding of canine leptospirosis is mainly based on serological tests that predict the infecting serogroup/serovar. Thus, the present study aimed at identifying the causative agent for severe cases of canine leptospirosis in a highly endemic area through the isolation and characterization of the isolated strains.ResultsUrine, serum and blood samples were collected from 31 dogs with suspected acute leptospirosis treated at the Veterinary Hospital Service of Santo Amaro University between 2018 and 2019. Acute infection was confirmed in 17 dogs (54.8%) by the associated use of Polymerase Chain Reaction (PCR), Microscopic Agglutination (MAT) and bacteriological culture. Eleven dogs (35.5%) had titers ≥800, with the most frequent serogroups being Autumnalis and Icterohaemorrhagiae (n = 4 each) and Canicola (n = 2). Leptospires were recovered from four dogs, and Multilocus Sequence Analysis (MLSA) revealed infection caused byL. interrogans, which were further characterized as serogroups Canicola (n = 1) and Icterohaemorrhagiae (n = 3).ConclusionThe identity of the isolates and serological pattern of MAT suggest that dogs are highly exposed to the serogroup Icterohaemorrhagiae and Canicola, also indicating possible circulation of serogroups not yet isolated in Brazil, notably serogroup Autumnalis. Our findings also reinforce the usefulness of using multiple diagnostic approaches to confirm acute canine leptospirosis.

Small Indian mongooses (SIMs, Herpestes auropunctatus) have invasively inhabited over 60 islands worldwide. They have been confirmed as a reservoir of rabies, leptospirosis, and salmonellosis; however, their role in the epidemiology of other zoonoses is little known. On St. Kitts, as well as other islands, SIMs harbor Ctenocephalides felis, which can vector several zoonotic diseases. In this study, SIMs were examined for fleas, and the collected fleas analyzed by PCR and DNA sequencing for Bartonella henselae, Rickettsia felis, Yersinia pestis, and Dipylidium caninum. Of the 87 SIMs, 75 (86.2%) harbored C. felis. C. felis recovered from nine (10.3%), one (1.1%), and one (1.1%) of the SIMs was positive for B. henselae, R. felis, and D. caninum, respectively. These data indicate that SIMs serve as an additional reservoir of B. henselae and R. felis, which should be taken into consideration in control and prevention of these rapidly emerging zoonoses.

Introduction Presentation of febrile illness with nonspecific features, overlapping manifestations of dengue and leptospirosis, limited laboratory diagnostic tests, make the clinical diagnosis of pyrexia a challenge. The present study aimed to determine the prevalence of Leptospira and dengue IgM co-infection among acute febrile illness patients. Methods This is a retrospective hospital-based study which included patient data collected from June 2016 to May 2017. Inpatients' samples (n=2139) were tested for dengue and/or Leptospira IgM at the Microbiology Laboratory. Data like duration of fever, platelet count, hemoglobin, white blood cell count, erythrocyte sedimentation rate, results of liver and renal function tests, mode of treatment, were collected from medical records of laboratory-confirmed co-infection cases. Results Among 1612 serum samples tested for dengue IgM by ELISA, 382 (23.7%) were positive, 17 equivocal and 1213 were negative. Of the 811 Leptospira IgM ELISA done, 119 (14.7%) were positive, 17 equivocal and 675 negative. Two hundred eighty-four samples were tested for both infections and nine (3.2%) were positive for both and 275 were negative. These nine patients positive for dual infections showed elevated transaminases, alkaline phosphatase, serum bilirubin, creatinine, and blood urea, thrombocytopenia and leukocytosis. They received effective antibiotics along with supportive treatment and were cured of the infection. Conclusions The study emphasizes the possibility of leptospirosis and dengue co-infection (3.2%) and need for confirmation by a highly specific test like PCR. If co-infection is suspected, treatment with specific antibiotics for leptospirosis and supportive treatment for dengue is mandatory, with due attention to complexity of organ involvement.

BACKGROUND: Leptospirosis or also commonly called Weil disease is an acute infectious disease that can attack humans and animals and is classified as a zoonotic disease. Leptospirosis is caused by bacteria of the genus Leptospira found in rodents. The difficulty of the diagnosis process causes the case of leptospirosis less reported and is one of the neglected infectious diseases. Patient findings are often not optimal because of underdiagnosis or misdiagnosis. AIM: This study aims to determine the presence of Leptospira serovar bacteria in blood serum of leptospirosis suspect using the microscopic agglutination test (MAT) method. METHODS: This study is a descriptive observational using accidental sampling technique. Sample collection was performed in the Antang Community Health Center, Batua Health Center, and Bangkala Health Center in Manggala District. RESULTS: This study found that there were 31 serum samples. Results of MAT examination on blood serum yielded 11 positive samples of Leptospira (37%) with serovar Hebdomadis, Djasiman, Mini, Icterohaemorrhagiae, Bangkingan, and Bataviae. CONCLUSIONS: The presence of Leptospira bacteria in blood serum of suspected individuals using the MAT method obtained 11 samples of positive blood serum. Special counseling activities are required about leptospirosis and diagnosis enforcement in the community or workers who experience symptoms or leptospirosis suspect.

Despite estimates suggesting Leptospira spp. being endemic in Southeast Asia, evidence remains limited. Diagnostic accuracy evaluations based on Leptospira ELISA mainly rely on hospitalized and severe patients; therefore, studies measuring the pathogen burden may be inaccurate in the community. We evaluated the Panbio Leptospira ELISA IgM among 656 febrile outpatients attending primary care in Chiangrai, Thailand, and Hlaing Tha Yar, Yangon, Myanmar. ELISA demonstrated limited diagnostic accuracy for the detection of acute leptospiral infection using the manufacturer recommended cutoff, with a sensitivity of 71.4% and specificity of 36.4%, and an area under the receiver operator characteristic curve value of 0.65 (95% CI: 0.41–0.89), compared with our reference test, the PCR assay. ELISA also performed poorly as a screening tool for detecting recent exposure to Leptospira spp. compared with the “gold-standard” microscopic agglutination test, with a specificity of 42.7%. We conclude that the utility of the Leptospira IgM ELISA for both serodiagnosis and seroprevalence is limited in our setting.

Leptospirosis is a zoonosis caused by pathogenic Leptospira spp. Cats have been reported to be infected with Leptospira spp. and shed the bacteria in the urine. However, the importance of cats as an infection source for humans remains unclear. In this study, Leptospira infection in cats in Okinawa Prefecture, Japan, where leptospirosis is endemic, was investigated by leptospiral antibody and DNA detection using microscopic agglutination test and nested PCR, respectively. Moreover, multilocus sequence typing (MLST) and whole genome sequencing (WGS) were conducted on the Leptospira borgpetersenii serogroup Javanica isolated from cats, black rats, a mongoose, and humans. Anti-Leptospira antibodies were detected in 16.6% (40/241) of the cats tested, and the predominant reactive serogroup was Javanica. The leptospiral flaB gene was detected in 7.1% (3/42) of cat urine samples, and their sequences were identical and identified as L. borgpetersenii. MLST and WGS revealed the genetic relatedness of L. borgpetersenii serogroup Javanica isolates. This study indicated that most seropositive cats had antibodies against the serogroup Javanica and that cats excreted L. borgpetersenii in the urine after infection. Further, genetic relatedness between cat and human isolates suggests that cats may be a maintenance host for L. borgpetersenii serogroup Javanica and a source for human infection.

Introduction Leptospirosis and rickettsial diseases are global zoonotic diseases. In severe infection cases, mortality can range from 10% to 30%. Currently most epidemiological data available are based on outbreak investigations and hospital-based studies from endemic countries. The U.S. soldiers at military bases in these countries are highly vulnerable due to the fact that most of them are immunologically naïve to these pathogens. No risk assessment of leptospirosis and rickettsial diseases among U.S. military personnel in Honduras is currently available. This study was aimed at determining the prevalence of leptospirosis and rickettsial diseases in U.S. military personnel deployed to Honduras using serological assays. Materials and Methods A cohort of pre- and post-deployment sera from the most recent 1,000 military personnel stationed in Honduras for at least 6 months between 2000 and 2016 was identified for this study. Serum specimens from these eligible subjects were retrieved. All post-deployment serum specimens were screened at a dilution of 1:100 for the presence of IgG antibodies to Leptospira and Rickettsia pathogens. The pre-deployment sera from those individuals with post-deployment IgG antibodies above cutoff (i.e., seropositive) were tested to determine seroconversion. Seroconversion was defined as conversion of an optical density value from below the cutoff (i.e., negative) in a pre-deployed specimen to above the cutoff (i.e., positive) in a post-deployed specimen at a titer of 100. Results The seropositive post-deployment specimens for antibodies against Leptospira (causing leptospirosis), Rickettsia typhi (causing murine typhus [MT]), spotted fever group rickettsioses (SFGR, causing SFG Rickettsia), Orientia tsutsugamushi (causing scrub typhus [ST]), and Coxiella burnetii (causing Q fever [QF]) were 11.6%, 11.3%, 6%, 5.6%, and 8.0%, respectively. The seroconverted rate in those assigned to Honduras from 2000 to 2016 was 7.3%, 1.9%, 3.9%, 4.3%, and 2.7% for leptospirosis, MT, SFGR, ST, and QF, respectively. Among the seroconverted specimens, 27 showed seroconversion of at least two antibodies. These seroconverted individuals accounted for 8.8% (3 out of 34) of the personnel who looked for medical attention during their deployment. Conclusions Our results suggest a leptospirosis seroconversion rate of 7.3%, which is higher than the 0.9% and 3.9% seroconversion in Korea and Japan, respectively. The higher rate of seroconversion indicates potential risk of Leptospira exposure. Additional testing of water samples in the pools and pits around the training sites to locate the infected areas is important to eliminate or reduce future exposure to Leptospira during trainings. The rates of seroconversion for ST, MT, spotted fever Rickettsia, and QF were 4.3%, 1.9%, 3.9%, and 2.7%, respectively, indicating the potential exposure to a variety of rickettsial-related pathogens. Testing of vectors for rickettsial pathogens in the areas could inform effective vector control countermeasures to prevent exposure. Proper precaution and protective measures are needed to better protect military personnel deployed to Honduras.

Over the period of research from 2003 to 2019 years, only 247 foci of leptospirosis of animals were registered in Yakutia, including: cattle – 61 (25.5 %), horses –159 (65.4 %), pigs –13 (5.3 %), small cattle – 8 (3.2 %), fur animals – 1 (0.4 %), bison – 4 (1.6 %). At the same time, a significant part of the foci of leptospirosis falls on horse breeding – 65.4 %. The largest number of disadvantaged points was established in 2008–49, including in horse breeding farms – 34 (69.4 %). An increase in the spread of the disease can be observed from 2003 to 2008 with a frequency of two years, and from 2008 – four years. Over the past 15 years, the introduction of leptospirosis pathogens has been established in 29 districts of the republic. It has been established that in the republic the main circulating serogroups of leptospirare Hebdomadis, Tarassovi, Icterohaemorrahagiae, Grippotyphosa, Canicola and Pomona. In zones of local economic stress, the source for the emergence and spread of leptospirosis are farms with a high number of farm animals. Of particular note are epizootic manifestations of leptospirosis in the Central Zone, which currently accounted for 80 % of the total quantity, in the remaining zones ranges from 2.5 to 7.5 %. It was revealed that the largest number of positive results was revealed in the spring period from April to June, and in the winter period – from November to December.

Background: In a highly globalized world, where boundaries are continually crossed due to different international activities, tropical diseases including Leptospirosis have emerged as significant causes of severe illness and death in untreated patients. Aim: To optimize the culture isolation of Leptospira by using different culture methods. Materials and Methods: All suspected cases of Leptospirosis regardless of their age/sex were recruited following informed and written consent. Five different blood culture methods have been tried to see better isolation of Leptospira from patient. Result: Total 326 patients are evaluated over a period of three years. Five different method of blood culture were followed out of that, whole blood culture technique shows better result compaire to other followed technique. Conclusion: The culture methods are gold standard but time consuming. It has an important role in the study of outbreaks. It provides epidemiological data that is helpful in studying pathogenesis of Leptospira. Keywords: Leptospirosis, Blood culture, Isolation, EMJH medium.

Background: Leptospirosis is most often diagnosed clinically, and a laboratory test with high diagnostic accuracy is required. Methods: IgM and IgG ELISAs using Leptospira antigens were established and evaluated in relation to the microscopic agglutination test (MAT). Antigen preparation consisted of saprophytic Leptospira biflexa to detect genus-specific antibodies (genus-specific ELISA) and a pool of the five most prevalent Leptospira interrogans serovars in Sri Lanka to detect serovar-specific antibodies (serovar-specific ELISA). IgM and IgG immune responses were studied in severe and mild leptospirosis patients (n = 100 in each group). Results: The ELISAs showed high repeatability and reproducibility. The serovar-specific IgM-ELISA showed a sensitivity of 80.2% and specificity of 89%; the genus-specific IgM-ELISA showed a sensitivity of 83.3% and specificity of 91%. The serovar- and genus-specific IgG-ELISAs showed sensitivities of 73.3% and 81.7%, respectively, and specificities of 83.3% and 83.3%, respectively. The commercial IgM-ELISA showed a sensitivity of 79.2% and specificity of 93%. The commercial IgG-ELISA showed a sensitivity of 50% and specificity of 96.7%. IgM levels observed in mild and severe leptospirosis patients were significantly higher than in the healthy control group, with mean absorbance values of 0.770, 0.778, and 0.163, respectively. Severe leptospirosis patients had significantly higher mean anti-leptospiral IgG levels compared to both mild leptospirosis patients and healthy control group subjects (0.643, 0.358, and 0.116, respectively; ANOVA, p < 0.001). The presence of anti-leptospiral IgG above an optical density of 0.643 at 1:100 could predict a high risk of severe disease. Conclusion: The serovar-specific in-house ELISA could be used for the laboratory diagnosis of leptospirosis in endemic settings. The high levels of anti-leptospiral IgG observed suggest its value as a predictor of disease severity.

Introduction. Leptospirosis is a zoonosis caused by microorganisms of the genus Leptospira with a global spread. The main reservoir of leptospires are the small mammals, which survive after infection, spreading the causative agent in the environment with urine. Material and methods. The studied were conducted in 2017-2019 in different ecosystems, where 1617 small mammals were collected, investigated for the presence of specific antibodies to Leptospira spp. Ecological analysis of small mammal communities was performed and leptospire-carrier species were identified. Results. 17 species of the order Rodentia and Soricomorpha were identified. The most widespread are A. sylvaticus, A. flavicollis and A. agrarius with a frequency of 100%, abundant and dominant in most of the studied ecosystems. Antibodies specific to Leptospira spp. were detected in 9 species, the highest share of the species with leptospire belongs to A. agrarius (53.85%) and C. glareolus (11.54%). The serogroups Leptospira grippotyphosa, L. icterohaemorrhagiae and L. pomona were highlighted. Conclusions. The epizootiological process in leptospirosis is maintained active in nature by 9 species of small mammals, including one of the Europe species A. agrarius, the leptospire rate was determined at 53.85%. The intense circulation of leptospires in the small mammal population determines the need to monitor the multiannual dynamics of their in order to establish changes in natural foci of leptospirosis, forecast the epizootological situation and the risk of spreading of disease among the human population.

The objective of this study was to evaluate the occurrence of antibodies against Leptospira spp in horses used by communities of informal waste collectors who use horse-drawn cartsin Guaíba, Rio Grande do Sul. Additionally, interviews were conducted with owners to assess the characteristics of the horses, their environment, and work. Serological testing for Leptospira spp. was performed on blood samples collected from 54 horses using the microscopic agglutination test. Results showed that 50.0% of the horses were seropositive for Leptospira spp and the most frequent serogroup was LeptospiraCanicola (88. 8%), followed by Leptospira Pyrogenes (11.1%), Leptospira Bataviae (7.4%), Leptospira Hebdomadis (7.4%), Leptospira Icterohaemorrhagiae (3.7%), Leptospira Australis (3.7%), Leptospira Pomona (3.7%) and Leptospira Copenhageni (3.7%). There were no statistically significant associations between seropositivity for Leptospira spp and age, gender, body score, water source, type of food storage, cohabitation with dogs, rodent infestation, residential flooding, and waste removal services. The prevalence of the Canicola serogroup, as well as a large number of dogs in close relationship with thehorses and the waste collectors, draws attention to the potential of these animals to become a source of Leptospira spp. infection in humans and other animals.

Rats are an important maintenance host of Leptospira spp., the causative agents of leptospirosis, a zoonotic disease with worldwide distribution. Rats are traded as food in the Mekong Delta, where Cambodia exports tons of rats to Vietnam. Handling wild rats is a potential health risk, but the information on Leptospira spp. carried by rats traded in the region remains limited. In this study, we investigated the carriage of Leptospira spp. in rats exported from Cambodia to Vietnam using bacterial culture, nested PCR and DNA sequencing. Isolates were then assessed using serological analysis and whole-genome sequencing (WGS), and Leptospira DNA detected in rat kidney tissues was also analysed by multilocus sequence typing (MLST). Seventy-two rats (2 Bandicota indica, 57 Rattus argentiventer, 11 R. losea, 1 R. norvegicus and 1 R. rattus) were subjected to bacterial culture, and three L. borgpetersenii serogroup Javanica isolates were obtained from R. argentiventer (5.3%). WGS revealed that although Cambodian isolates were genetically related to L. borgpetersenii serogroup Javanica strains widely distributed in East and South-East Asian countries, they formed a different cluster from other strains. In addition to the three L. borgpetersenii sequences, the flaB sequence of L. interrogans was detected in 18 R. argentiventer and 7 R. losea kidney tissue samples (38.9%) using nested PCR followed by DNA sequencing. The L. interrogans flaB-positive samples were further analysed by MLST, revealing that seven housekeeping genes (glmU, pntA, sucA, tpiA, pfkB, mreA and caiB) contained novel sequences with distinct lineages from other sequence types. This study revealed a high prevalence of Leptospira spp. among rats exported from Cambodia to Vietnam, indicating a potential risk to people engaging in rat trade and demonstrating that a fastidious L. interrogans strain circulates among Cambodian rats.

The authors analyzed the main indicators of nosological profile and epizootic manifestations of infectious diseases of pigs in the Krasnodar territory for the period from 1990 to 2019. As a result of the analysis of pig diseases of infectious etiology, 24 nosological forms of diseases were established. During the study period, 3006 disadvantaged areas were registered in the province, where 472.6 thousand heads of pigs of various age and sex groups fell ill, of which 131.3 thousand fell ill. In the nosological structure of infectious diseases of pigs, bacterial, viral and chlamydial infections occupy a significant share: colibacteriosis-34.0 %, salmonellosis-20.3, pseudomonosis-9.9, streptococcosis-7.9, pasteurellosis-5.6, viral gastroenteritis-4.2, erysipelas-3.4, African swine fever-2.3, chlamydia-0.5 %. The largest number of points that were affected by infectious diseases of pigs was registered in 1999 - 354; of these, 4.8% were viral diseases: aujeski disease, viral gastroenteritis; 94.9 - for bacterial: streptococcosis, diplococcosis, colibacteriosis, edema, leptospirosis, pasteurellosis, salmonellosis, erysipelas, pseudomonosis; 0.3 % - for chlamydia. The smallest number of points with problems related to infectious diseases of pigs was registered in 2013: in 1 point with problems related to African swine fever (ASF), 3 heads fell ill. The number of sick animals was the highest in 1994: 66.9 thousand animals fell ill in 80 disadvantaged areas, of which 23.4 thousand died. The greatest economic damage to pig farms was caused by ASF. This infection led to a reduction in the number ofpigs in the Krasnodar territory from 1200.0 to 300.0 thousand heads. In the period from 2008 to 2019, ASF was registered on the territory of the province in 33 out of 44 municipalities, in 79 disadvantaged localities, including 13 among the wild boar population. In total, 7.4 thousand heads of domestic pigs and 157 wild boars fell ill in the centers of ASF, 33 objects infected with the ASF virus were identified.

The most common presentation of animal leptospirosis is the subclinical and silent chronic form, that can lead to important reproductive disorders. The diagnosis of this chronic form remains a challenge. The aim of the present study is to gather and critically analyse the current information about molecular tools applied to animal leptospirosis diagnosis, particularly the silent chronic presentation of the infection. Regarding clinical specimens, samples from urinary tract were the most used (69/102, 67·7%), while few studies (12/102, 11·8%) investigated samples from reproductive tract. Concerning the molecular methods applied, the most used is still the conventional polymerase chain reaction (PCR) (46/102, 45%), followed by real-time PCR (38/102, 37·2%). The lipL32 gene is currently the most common target used for Leptospira detection, with 48% of studies applying this genetic marker. From all the studies, only few (21/102, 20·5%) performed gene sequencing. According to the majority of authors, current evidence suggests that lipL32-PCR is useful for an initial screening for Leptospira DNA detection in animal clinical samples. Posteriorly, if DNA sequencing could be performed on positive lipL32-PCR samples, we encourage the use of secY gene as a genetic marker. The molecular methods appear as the most important tools for the diagnosis of the chronic silent leptospirosis on domestic animals, reinforcing its evident impact not only on animal reproduction but also on a One Health context.

Background Leptospirosis is a zoonotic disease of concern and an investigation of recent spatio-temporal trends of leptospirosis in dogs in the United States is needed. Leptospira PCR testing has become increasingly used in veterinary clinical medicine and these data might provide information on recent trends of disease occurrence. Objectives To identify and describe clusters of PCR-positive Leptospira test results in dogs in the United States. Animals Leptospira real-time PCR test results from dogs (n = 40 118) in the United States from IDEXX Laboratories, Inc., between 2009 and 2016 were included in the analysis. Methods In this retrospective study, spatio-temporal clusters for a real-time PCR-positive test were identified using the space-time permutation scan statistic and the centroid of the zip code reported for each test. A maximum spatial window of 20% of the population at risk, and a maximum temporal window of 6 months were used. Results Seven statistically significant space-time clusters of Leptospira real-time PCR-positive test results were identified across the United States: 1 each located within the states of Arizona (2016), California (2014-2015), Florida (2010), South Carolina (2015), and 1 each located within the south-central region (2015), midwest region (2014), and northeast region (2011). Clusters ranged from 3 to 108 dogs and were identified during all years under study, except 2009, 2012, and 2013. Conclusions and clinical importance The spatial and temporal components of leptospirosis in dogs in this study are similar to those in previous work. However, clusters were identified in new areas, demonstrating the complex epidemiology of this disease.

Objectives Recent studies show that cats could play an important role in the transmission of Leptospira species. There are few reports of leptospirosis on Prince Edward Island (PEI) and none in cats. The objective of this study was to determine the prevalence of serum antibodies against Leptospira serovars and of Leptospira DNA in the urine of a population of free-roaming cats. Methods Paired blood and urine samples were collected from 200 cats brought to a trap–neuter–return program. Antibody titers against six Leptospira serovars (Bratislava, Canicola, Gryppotyphosa, Hardjo, Pomona, Icterohaemorrhagiae) were determined by microscopic agglutination test. PCR was performed on urine samples to identify urine shedding of Leptospira DNA. Results Antibodies were detected in 20/200 cats (10%) for at least one serovar, with titers ranging from 1:50 to 1:6400 (all serovars tested, except Hardjo). Urine samples of 7/200 cats (3.5%) were PCR-positive. Conclusions and relevance Feral cats in PEI had a higher than expected exposure to leptospirosis and can shed DNA from pathogenic Leptospira species in urine. Further studies are needed to determine the prevalence of exposure to leptospirosis in other species on PEI and the potential role of feral cats in transmission of the disease.

Introduction: FUO/ PUO (Fever/Pyrexia of Unknown Origin) is referred to when temperature is observed above 38.30C (1010F) on many occasions over a period of > 3 weeks and unable to diagnose despite 1week of thorough investigations. Different studies reported diagnosis of malaria in 5 to 50% cases; leptospirosis in 3 to 10% cases and influenza in 8 to 12% cases Dengue fever and malaria are arthropod born diseases and endemic in many parts of India during the monsoon season. Leptospirosis and scrub typhus are zoonotic infections and are widely prevalent in areas with heavy monsoon and agrarian way of life. Aim: To evaluate the study of various Rickettsia infections in patients suffering from Fever Of Unknown Origin. Objectives: To understand the occurrence of infections caused by rickettsial species in suspected cases of FUO. To increase awareness and clinical suspicion among doctors for these infections. Materials and Methods: The assay was performed using P.vulgaris OX19, OX2, OXK strains according to manufacture’s instructions by tube method. Serum will be diluted from 1/20 to 1/640 titer. A singleWeil Felix titer of >1:160 or fourfold rise in titers on repeat testing starting from 1:40 will be accepted as a positive result. Result: A total 100 cases were including in my study in which 28 cases were positive. Out of 28 positive rickettsial samples, females were 17 and males were 11. Spotted Fever(9), Endemic Fever(8), Epidemic Fever(7), Scrub Typhus Fever(4). Conclusion: With the growing number of cases detected in India, scrub typhus is fast emerging as a public health threat and also due to limited diagnostics leading to underreporting, Weil Felix test could be used in adjunct with Enzyme-linked immunosorbent assay and blood parameters in the diagnosis of rickettsial diseases. Keywords: Fever/Pyrexia of unknown Origin, (FUO/PUO), Spotted fever, Endemic fever, Epidemic fever, Scrub typhus fever, Weil felix.

Leptospirosis caused by Leptospira interrogans is considered as one of the most important zoonotic infections globally. It affects humans and wide range of animals. It damages various vital organs of the body and can often lead to fatal complications. The various symptoms of leptospirosis are fever, headache, nausea, vomiting and abdominal pain. It may lead to complications like jaundice, hemorrhage, myocarditis, meningitis and in some cases renal failure. Leptospirosis is mainly caused by rodents which are considered as reservoir hosts for leptospires. The other animals such as mammals, birds, amphibians, reptiles and fish are carriers of leptospirosis. Humans acquire leptospirosis accidently by contact with carrier animals or environment contaminated by leptospires. Leptospirosis has a wide geographical distribution including tropical, subtropical and temperate climatic regions. But it is more prevalent in the tropical areas where warm humid conditions and alkaline or neutral soil helps leptospires survive better. This review is an attempt to cover every aspect of leptospirosis in detail. The biology and culture characteristics of leptospires, classification, epidemiology, pathogenesis, host immune response, transmission cycle and different diagnostic methods have been explained in detail.

Studies on caprine leptospirosis using isolation, histochemistry and immunohistochemistry are rare. The role of small ruminants in the epidemiology of leptospirosis is scarce. This study investigated the prevalence of Leptospira spp. serovars, and the renal pathology of caprine leptospirosis in slaughterhouses from two states in southwest Nigeria using isolation (IS), Warthin Starry silver (WSs) impregnation and immunohistochemistry (IH). One hundred and sixty-nine kidney samples were randomly obtained from goats between September 2015 and June 2017. Chi-square test was used with a confidence level set at 0.05 to ascertain associations between the positive cases, sex and animal species. Eighty-seven (51.5%) samples were positive on IS, out of which 26/40 and 25/30 were positive on WSs and IH, respectively. Ten (5.9%) kidneys showed macroscopic lesions while interstitial nephritis (48.6%) and tubular nephrosis (64.2%) were the most prominent histopathological changes. The most frequently observed positive reactions were against serovars Hardjo type Prajitno (12/25, 48%), and Gripptotyphosa (5/25, 20%). Other serovars such as Bratislava (2/25, 8%), Canicola (3/25, 12%), Icterohaemorrhagiae (2/25, 8%), and Pomona (1/25, 4.0%) were also detected using IH. The result showed high prevalence of Leptospira infection in goats and the possibility of humans contracting the disease. To date, the detection of leptospirosis from kidneys of goats using IS, WSs and IH has not been reported. This study is the first documentation of evidence of pathogenic Leptospira species in renal tissues of goats.

Leptospirosis is a widespread zoonosis in Russia. The human leptospirosis vaccine produced by Rostov Research Institute of Microbiology and Parasitology has been used since 1998 to minimise risks associated with deterioration of epidemiological situation. Lately, there has been an increase in the incidence of leptospirosis caused by Leptospira interrogans serovar Canicola which is not included in the human vaccine. The aim of the study was to analyse biological properties of Leptospira interrogans serovar Canicola strains to substantiate their inclusion into human leptospirosis liquid concentrated inactivated vaccine. Materials and methods: two L. interrogans serovar Canicola strains: Udalov 480 and Sobaka 2000 were used in the study. Virulent properties of the strains were evaluated by infecting Syrian hamsters, the data on specific activity of the experimental vaccine were compared to specific activity of the leptospirosis liquid concentrated inactivated vaccine used for disease prevention. The experimental vaccine was tested for Specific safety and Abnormal toxicity according to FS.3.3.1.0014.15 Leptospirosis liquid concentrated inactivated vaccine. Results: the comparison of Udalov 480 and Sobaka 2000 strains of L. interrogans serovar Canicola revealed higher virulence of Udalov 480. The tests performed for the experimental vaccine batches demonstrated that the inclusion of Udalov 480 strain did not affect the above-mentioned properties of the vaccine. Conclusions: the study demonstrated the possibility of using L. interrogans serovar Canicola strain Udalov 480 as a component of the currently produced vaccine, and confirmed the safety and efficacy of the new vaccine.