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• DPP for diagnosing leptospirosis was evaluated in reference laboratories in Brasil. • The test was less sensitive than specific. • The highest accuracy occurred between 7 and 14 days after symptom onset. • Accuracy measures were similar between the laboratories. • Predictive values indicated the best situations to utilize DPP in health services. Leptospirosis is a zoonotic disease with significant global impact and a challenging diagnosis. The utilization of adequately validated rapid tests is relevant for the opportune identification of the disease and for reduction in fatality rates. The present study analyzes the accuracy and reliability of the Dual Path Platform (DPP) assay -produced in Brazil by the Oswaldo Cruz Foundation (Fiocruz)- for diagnosing leptospirosis. Firstly, a serological panel was constructed in the Brazilian Reference Laboratory for Leptospirosis using samples routinely handled by reference laboratories of six Brazilian states. It consisted of 150 positive (according to MAT and IgM-ELISA) and 250 negative samples for leptospirosis. Subsequently, the panel samples were distributed to the reference laboratories for the performance of DPP assays in triplicate. Different measures were used in the assessment of diagnostic quality. Predictive values were estimated for different pre-test probability settings. Sensitivities varied between 67.33 % and 74.00 % and specificities between 93.20 % and 98.40 % in the states, and there were adequate agreements between them. Accuracies were lower for the samples of patients with less than 7 days of symptoms. In contexts of prevalence values up to around 25 %, positive and negative predictive values were around 90 %. However, in situations of high pre-test probabilities, NPVs were low. This study improves understanding of the use of DPP in diagnosing leptospirosis, particularly its application in healthcare settings. As long as the time of symptoms onset and clinical and epidemiological contexts are adequately considered for the interpretation of results, DPP is a valid option to be used in the leptospirosis diagnostic routine. [ABSTRACT FROM AUTHOR]

• Equine leptospirosis is an infectious disease with a worldwide distribution, mainly in tropical areas. • Horses are particularly susceptible to the disease, presenting manifestations ranging from subclinical to the development of uveitis. • Blood and urine samples were collected from 34 horses and one mule from three localities in the state of Veracruz, Mexico. • The results reflected an overall serological frequency of 97.1 %. Additionally, we recovered 23 urine isolates. • Our results represent the first record of the presence of Leptospira interrogans and Leptospira santarosai through bacteriological isolates in equids from Mexico. Leptospirosis is an infectious disease with a worldwide distribution, which represents a major challenge in animal production across developing countries, mainly in tropical areas. Horses are particularly susceptible to the disease, presenting manifestations ranging from subclinical to the development of uveitis that compromises the visual health of the animals. In recent years, serological studies have been carried out in equid populations from America, demonstrating high exposure. For this reason, the aim of this study was to demonstrate microbiologically and molecularly the presence of the members of the genus Leptospira in urine samples from equids in an endemic state of leptospirosis in Mexico, and to detect the serological presence of anti- Leptospira antibodies in the sampled animals. For this reason, blood and urine samples were collected from 28 horses and one mule from three localities in the state of Veracruz, Mexico. Urine samples were inoculated in Ellinghausen–McCullough–Johnson–Harris (EMJH) medium, and the recovered isolates were typed using a short Multi Locus Sequence Typing scheme. Amplifications of the expected size were subjected to sequencing, and the recovered sequences were compared with those of reference deposited in GenBank using the BLAST tool. To identify their phylogenetic position, we performed a phylogenetic reconstruction using the maximum likelihood method. Additionally, Microscopic Agglutination test was performed on the serum samples to identify anti- Leptospira antibodies. We recovered 16 urine isolates which tested positive for the presence of Leptospira DNA. The phylogenetic reconstruction and the MLST analysis confirmed the presence of several genotypes of Leptospira interrogans and Leptospira santarosai. An overall serological frequency of 97.1 % was detected. Our results represent the first record of the presence of Leptospira through bacteriological isolates in equids from Mexico. [Display omitted] [ABSTRACT FROM AUTHOR]

• Four pathogenic Leptospira from animal origin were characterized by MLST approach. • Eighteen different MLST sequence types (STs) were found (22 new alleles and 11 new STs). • The Leptospira santarosai population demonstrated intraspecific variability as high as the interspecific of Leptospira spp. In the current context of the emergence of certain infectious diseases and discussion of the One Health concept for many of these, the study of leptospirosis – both in domestic and wild hosts – cannot be neglected. The study of animal leptospirosis has evolved in recent years. It has been demonstrated that the human-animal-environment interface is more important than previously thought. In the present study, 35 strains of five pathogenic Leptospira species were isolated from different animal species in Brazil and characterized by rrs, sec Y, and Multilocus Sequence Typing (MLST) sequencing. Phylogenetic inferences were performed and the molecular diversity of the populations (intra- and inter-population levels) was evaluated. Among the five studied species, 18 different sequence types (STs) were found (22 new alleles and 11 new STs). eBURST analysis revealed two clonal complexes (CCs) and seven singletons. A high genetic diversity was demonstrated (H = 0.954 ± 0.017), mainly for the L. santarosai population (H = 0.942 ± 0.034, n = 20). The same strain was identified in different host species, as well as strains with zoonotic potential circulating in the country. Although the difficulty of culturing Leptospira strains is well known, the high variability of the strains found in Brazil highlights the importance of animals in maintaining the biological cycle of the bacterium in nature. Moreover, the selection of autochthonous strains for the development of vaccines becomes a challenge. [ABSTRACT FROM AUTHOR]

• This is the first reported detection of pathogenic leptospires in urine of domesticated elephants in Sri Lanka. • Of the 13 elephants, 4 were shedding Leptospira interrogans. • Domestic elephants are in close contact with the public in Sri Lanka pose a risk of transmitting leptospirosis to humans. Leptospirosis is a globally common zoonotic infectious disease in humans and animals. This disease is caused by pathogenic spirochetes belonging to the genus Leptospira. The pathogen is able to survive in mammalian kidneys after infection and is excreted in urine intermittently. Pathogenic leptospires infect humans either by direct contact with infected animal urine or through contaminated soil or water. In Sri Lanka, some studies have demonstrated the involvement of animals, such as livestock species and peridomestic rats, in the transmission of leptospirosis to humans. However, none of the previous studies focused on domesticated elephants, which are in close contact with humans during cultural and religious events and bathe in rivers together with humans. If domesticated elephants act as carriers of pathogenic leptospires, it could be a major public health issue in the country. In this study, 13 healthy domesticated elephants were subjected to leptospiral DNA detection from urine samples collected on three consecutive days. Four elephants (31%) were confirmed to shed pathogenic leptospires in their urine. DNA sequencing followed by phylogenetic distance measurements revealed that all positive elephants were infected with L. interrogans. This study reveals the possibility that elephants act as a source of infection of leptospires to humans and recommends the screening of all domesticated elephants that are in close contact with humans for the shedding of pathogenic leptospires. [ABSTRACT FROM AUTHOR]

Abstract Swine leptospirosis poses a major problem in the agricultural sector. The gold standard for serodiagnosis of leptospirosis is Microscopic Agglutination Test (MAT). However, the limitations of this technique make the search for alternative diagnostic methods inevitable. In the present study, ErpY-like recombinant protein (rErpY-like), produced in Escherichia coli and used as antigen in indirect enzyme-linked immunosorbent assay (ELISA), was evaluated for its efficacy as a novel diagnostic tool for swine leptospirosis. For the study, 72 samples of swine sera characterized by microscopic agglutination test (MAT), were evaluated by indirect ELISA. The sensitivity, specificity and accuracy values obtained from the analysis were 96.8%, 100%, and 99%, respectively, thereby suggesting that rErpY-like ELISA being a sensitive and specific method for antibodies detection in swine populations could be used as an alternative for diagnosis of swine leptospirosis. [ABSTRACT FROM AUTHOR]

Abstract The present study aimed to detect the most prevalent serogroups and circulating Leptospira species in cows from Brazilian Amazon. Samples of blood serum, urine and kidney of 208 animals were collected at a municipal slaughterhouse in the Baixo Tocantins region of Pará State, Northern Brazil. The tests used were microscopic agglutination test (MAT), bacteriological isolation, polymerase chain reaction (PCR) and DNA sequencing. The frequency of MAT-reactive cows was 46.6% (97/208) with titers ranging from 100 to 3200, being Sejroe serogroup the most prevalent. There was no Leptospira isolation, but the DNA of bacterium was detected in 5.8% (12/208) of the kidney and in 14.9% (31/208) of the urine samples. DNA sequencing was performed directly from PCR products of 30 samples (3 kidneys and 27 urines), with identification of four different species: L. borgpetersenii with 56.7% (17/30), followed by L. kirschneri with 13.3% (4/30), L. interrogans with 6.7% (2/30), L. santarosai with 3.3% (1/30), and 20.0% (6/30) of samples were identified only at the genus level. These results reveal a diversity and peculiarity for bovine leptospirosis in the Amazon region, mainly due to the low frequency of L. santarosai and more surprising, the presence of L. kirschneri , differently of what is observed in other regions of Brazil. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Leptospira detection of 14.3% was recorded among the 266 small mammals captured. • R. norvegicus , R. rattus , M. whiteheadi , S. muelleri , T. gliss , R. tiomanicus and S. murinus carried pathogenic leptospires. • ST110 L. kirschneri Grippotyphosa, ST242 L. weilii , ST50 L. interrogans Bataviae ST143 L. borgpetersenii Javanica were detected. • To include the isolated local strains into MAT panel to enhance the leptospirosis diagnostic efficacy in Malaysia. Abstract Leptospirosis is caused by the spirochetal bacterium Leptospira of which rodents are considered the most important reservoir. This study aims to determine and characterize virulent Leptospira species among rodents and small mammals found in human settlements and recreational spots within the Hulu Langat and Gombak districts of Selangor, Malaysia; regions that frequently report probable human leptospirosis cases. Molecular analysis revealed an overall Leptospira detection rate of 14.3% among the 266 small mammals captured, and the human settlements were found to have the highest number of isolates (15.1%), followed by recreational sites (14.5%). The molecular characterization conducted based on the lipL32 , secY genes and MLST revealed that the strains belonged to four different species, including; Leptospira interrogans (29; 76.3%; ST50, ST238, ST243), L. kirschneri (5; 13.15%; ST110), L. borgpetersenii (3; 8%; ST143) and L. weilii (1; 2.63%; ST242). The study revealed genotypes of circulating strains among small mammals in Malaysia, which include Leptospira locus ST110 L. kirschneri, ST 50 L. interrogans , ST143 L. borgpetersenii and ST242 L. weilii. Among the small mammals studied, 17/105 (16.2%) Rattus norvegicus , 7/59 (11.9%) of Rattus rattus , 5/24 (20.8%) of Maxomys whiteheadi , 4/18 (22.2%) of Sundamys muelleri , 2/22 (9%), Tupaia gliss , 2/16 (12.5%) Rattus tiomanicus and 1/4 (25%) of Suncus murinus carried pathogenic leptospires. The data from the present study may imply that, in addition to rodents, other small mammals also serve as maintenance hosts for Leptospira. Hence, much remains unknown about Leptospira maintenance hosts, and there is need for further investigation to ascertain the prevailing serovars of pathogenic Leptospira in Malaysia. This will assist in the development of efficient diagnostic assays with improved microscopic agglutination test (MAT) panels, and in the implementation of suitable prevention and control measures. [ABSTRACT FROM AUTHOR]

Abstract Leptospirosis is a zoonotic disease that poses public health and ecological threats worldwide. In New Zealand (NZ), incidence of the disease is relatively high compared to other developed countries. The aim of this review was to describe the epidemiological status, ecological risk of leptospirosis and prevention in NZ. Disease notification data for leptospirosis in humans in NZ from 2010 to 2015 were collected from the Institute of Environmental Science and Research database. These data were supported by a literature review of epidemiological studies in human and animal populations. During this six-year period, exotic serovars of Leptospira interrogans sv Australis and Leptospira kirschneri sv Grippotyphosa were identified in patients who had travelled abroad to Samoa (Pacific Ocean) and Thailand, respectively. Most cases of leptospirosis were recorded in New Zealanders of European ethnicity, followed by Māori people. Males had a nine-fold increased risk compared to females mostly due to occupation. The risk of leptospirosis increased gradually with the age, with a peak in the 40 to 49-year-old age group, after which it decreased. Workers in meat-processing and farming industries appeared at highest-risk of occupational exposure compared with other risky occupations such as hunters, veterinarians, technicians, stock truck drivers and lake workers. Other cases were also attributed to outdoor exposures or travelling overseas. Highest disease notification rates occurred in the West Coast region of the South Island (average annual incidence 9.7 per 100,000 people), followed by Whanganui region (8.2 per 100,000) and Hawke's Bay region (8 per 100,000) in the North Island. Vaccines currently available for animals are specific for cattle, sheep, deer and dogs and do protect against all serovars present in NZ. The development and use of molecular diagnostics is crucial for specific identification of Leptospira isolates and informing deployment of efficient vaccines. [ABSTRACT FROM AUTHOR]

Vaccination is one of the most important measures for the control of bovine leptospirosis. Despite the broad usage of vaccination against leptospirosis in cattle worldwide, the dynamics of the post-vaccine immune response remain controversial and many aspects are still unclear, particularly in naturally-infected animals. Thus, the objective of this study is to describe the dynamics of humoral response in naturally-infected cattle after vaccination against leptospirosis. A total of 162 cows were studied, consisting of 129 included in the experimental group (G1), and subdivided into two groups, vaccinated with two different brands of bacterins, as well as 33 in the control group (G2). Serology (MAT) was performed in all cows on D0 (vaccination), then 60 and 120 days post-vaccination. Vaccination significantly elicited the production of anti-leptospiral antibodies. Seroreactivity increased rapidly but was of short duration (up to D60). Significantly, that increase was notably higher in the vaccinated group than in the controlled. Both vaccines elicited a similar response with a higher rate of seroreactive animals, but predominately against different serogroups. In this context, our results reinforce that, although of limited duration, vaccination against leptospirosis significantly elicits a specific humoral response in naturally-infected animals. The two studied vaccines presented similar seroconversion levels, but predominantly to different serogroups, being one against Icterohaemorrhagiae and the other against Sejroe. [ABSTRACT FROM AUTHOR]

Leptospirosis is a worldwide emerging infectious disease of zoonotic importance and large epidemics and epizootics have been reported all over the globe. A cross survey study was conducted to estimate seroprevalence of human leptospirosis in climatically distinct regions of Pakistan and to identify the risk factors associated with the disease. Blood samples from 360 humans were collected through convenient sampling, 120 from each of three study areas. Serological testing was performed using ELISA kit as per manufacturer’s recommendations. The results showed an overall prevalence of 40.83% (95% CI; 35.71–46.11). Statistical analysis showed significant (P < .05) differences in leptospiral seroprevalence in three different geographic locations, with highest in humid sub-tropical climatic region (50.83%; 95% CI; 41.55–60.07), followed by semi-arid region (44.16%; 95% CI; 35.11–53.52) and lowest in hot and dry region (27.50%; 95% CI; 19.75–36.40). After multivariate analysis age, gender, exposure to flooding water, source of water usage, disinfection schedule of surroundings and history of cut and wound were found significantly associated with the seropositivity of Leptospira . The present study, first to uncover seroprevalence of human Leptospira in different climatic regions of Pakistan, alarms about effect of climate on prevalence of Leptospira in the region. [ABSTRACT FROM AUTHOR]

In Iran, leptospirosis is endemic to Caspian Sea littoral. The disease appears as a seasonal infection mostly affecting people in rural areas involved in farming. We investigated the prevalence of this infection among suspected patients in Gilan Province by an indirect immunofluorescent assay (IFA), and two PCR protocols, a nested-PCR and a real-time PCR (qPCR), targeting rrs and lipL32 genes, respectively. We also identified the common Leptospira species by sequencing a partial sequence of rrs gene. Out of the 128 sera examined by IFA, 25.78% were positive with the antibody titers ≥1/80. The antibody titer in 39.06% of sera ranged from 1/10 to 1/140, and 35. 16% showed no antibodies, all considered negative. Nested PCR and qPCR detected Leptospira DNA in 20.31% and 18.75% of the sera, respectively. The two PCR assays had 98.43% agreement ( K  = 0.93) and showed an inverse correlation with the IFA titers. Also, three pathogenic Leptospira species, L . kirschneri (n = 10),  L. introgans (n = 8), and L. borgpetersenii (n = 2) were identified from the clinical specimens in the study area. In our hands both PCR assays proved very efficient for early diagnosis of illness and could be used in combination with IFA for both diagnosis and epidemiological studies, but nested PCR was cheaper and appeared more appropriate for our laboratories in rural settings. [ABSTRACT FROM AUTHOR]

The occurrence of tick-borne relapsing fever and leptospirosis in humans in Madagascar remains unclear despite the presence of their potential vectors and reservoir hosts. We screened 255 Amblyomma variegatum ticks and 148 Rhipicephalus microplus ticks from Zebu cattle in Madagascar for Borrelia -specific DNA. Borrelia spp. DNA was detected in 21 Amblyomma variegatum ticks and 2 Rhipicephalus microplus ticks. One Borrelia found in one Rhipicephalus microplus showed close relationship to Borrelia theileri based on genetic distance and phylogenetic analyses on 16S rRNA and flaB sequences. The borreliae from Amblyomma variegatum could not be identified due to very low quantities of present DNA reflected by high cycle threshold values in real-time-PCR. It is uncertain whether these low numbers of Borrelia spp. are sufficient for transmission of infection from ticks to humans. In order to determine whether spirochaete infections are relevant in humans, blood samples of 1009 patients from the highlands of Madagascar with fever of unknown origin were screened for Borrelia spp. − and in addition for Leptospira spp. − by real-time PCR. No target DNA was detected, indicating a limited relevance of these pathogens for humans in the highlands of Madagascar. [ABSTRACT FROM AUTHOR]

The bovine leptospirosis is an economically important zoonotic disease of flood affected areas worldwide, but scarce information is available about its epidemiology in Pakistan. This is a first study on sero-epidemiology of bovine leptospirosis in Pakistan. The objectives of this study were to investigate the sero-prevalence and associated risk factors of bovine leptospirosis in flood affected zone of Punjab, Pakistan. A total of 385 serum samples were randomly collected from four tehsils of district Muzaffargarh, Pakistan. The serum samples were subjected to indirect ELISA for the detection of anti-leptospira antibodies. The overall sero-prevalence of leptospirosis was 30.39%. The prevalence was significantly higher ( p < 0.001) in cattle (56.25%) than buffaloes (4.66%). The key risk factors identified based on multivariable logistic regression were; confined system of rearing, flooded area, and lesser to graduate level of education as risk factors for leptospirosis. The values of hematological parameters varied significantly ( p < 0.05) for Hb, TEC and PCV while there was no significant ( p > 0.05) difference among TLC values among sero-positive and sero-negative animals. The serum biochemical profile revealed significant differences ( p < 0.05) in values of ALT, AST and creatinine among the sero-positive and negative animals. Hence, it can be concluded that leptospirosis is an emerging and neglected disease in flood affected zone of Pakistan, and the disease needs to be explored comprehensively in other parts of the country to sort out solid strategies for its control and eradication. [ABSTRACT FROM AUTHOR]

We evaluated the renal carriage of Leptospira species in rodent communities from Mediterranean Chile using a PCR technique. We found that animals inhabiting agricultural areas were almost three times more infected than in wild areas (14.4% vs . 4.4%). The Norwegian rat ( Rattus norvegicus ), an invasive murid ubiquitous in the country, was the most infected species (38.1%). [ABSTRACT FROM AUTHOR]

Immuno-pathogenesis of leptospirosis can be recounted well by following its trail path from entry to exit, while inducing disastrous damages in various tissues of the host. Dysregulated, inappropriate and excessive immune responses are unanimously blamed in fatal leptospirosis. The inherent abilities of the pathogen and inabilities of the host were debated targeting the severity of the disease. Hemorrhagic manifestation through various mechanisms leading to a fatal end is observed when this disease is unattended. The similar vascular destructions and hemorrhage manifestations are noted in infections with different microbes in endemic areas. The simultaneous infection in a host with more than one pathogen or parasite is referred as the coinfection. Notably, common endemic infections such as leptospirosis, dengue, chikungunya, and malaria, harbor favorable environments to flourish in similar climates, which is aggregated with stagnated water and aggravated with the poor personal and environmental hygiene of the inhabitants. These factors aid the spread of pathogens and parasites to humans and potential vectors, eventually leading to outbreaks of public health relevance. Malaria, dengue and chikungunya need mosquitoes as vectors, in contrast with leptospirosis, which directly invades human, although the environmental bacterial load is maintained through other mammals, such as rodents. The more complicating issue is that infections by different pathogens exhibiting similar symptoms but require different treatment management. The current review explores different pathogens expressing specific surface proteins and their ability to bind with array of host proteins with or without immune response to enter into the host tissues and their ability to evade the host immune responses to invade and their affinity to certain tissues leading to the common squeal of hemorrhage. Furthermore, at the host level, the increased susceptibility and inability of the host to arrest the pathogens’ and parasites’ spread in different tissues, various cytokines accumulated to eradicate the microorganisms and their cellular interactions, the antibody dependent defense and the susceptibility of individual organs bringing the manifestation of the diseases were explored. Lastly, we provided a discussion on the immune trail path of pathogenesis from entry to exit to narrate the similarities and dissimilarities among various hemorrhagic fevers mentioned above, in order to outline future possibilities of prevention, diagnosis, and treatment of coinfections, with special reference to endemic areas. [ABSTRACT FROM AUTHOR]

Leptospirosis is a widespread zoonotic disease caused by pathogenic Leptospira species (Leptospiraceae). LipL32 is an abundant lipoprotein from the outer membrane proteins (OMPs) group, highly conserved among pathogenic and intermediate Leptospira species. Several studies used LipL32 as a specific gene to identify the presence of leptospires. This research was aimed to study the characteristics of LipL32 protein gene code, to fill the knowledge gap concerning the most appropriate gene that can be used as antigen to detect the Leptospira . Here, we investigated the features of LipL32 in fourteen Leptospira pathogenic strains based on comparative analyses of their primary, secondary structures and 3D modeling using a bioinformatics approach. Furthermore, the physicochemical properties of LipL32 in different strains were studied, shedding light on the identity of signal peptides, as well as on the secondary and tertiary structure of the LipL32 protein, supported by 3D modelling assays. The results showed that the LipL32 gene was present in all the fourteen pathogenic Leptospira strains used in this study, with limited diversity in terms of sequence conservation, hydrophobic group, hydrophilic group and number of turns (random coil). Overall, these results add basic knowledge to the characteristics of LipL32 protein, contributing to the identification of potential antigen candidates in future research, in order to ensure prompt and reliable detection of pathogenic Leptospira species. [ABSTRACT FROM AUTHOR]

Background Additive Bayesian Network (ABN) is a graphical model which extends Generalized Linear Modelling (GLM) to multiple dependent variables. The present study compares results from GLM with those from ABN analysis used to identify factors associated with Leptospira interrogans sv Pomona (Pomona) infection by exploring the advantages and disadvantages of these two methodologies, to corroborate inferences informing health and safety measures at abattoirs in New Zealand (NZ). Methodology and findings In a cohort study in four sheep slaughtering abattoirs in NZ, sera were collected twice a year from 384 meat workers and tested by Microscopic Agglutination with a 91% sensitivity and 94% specificity for Pomona. The study primarily addressed the effect of work position, personal protective equipment (PPE) and non-work related exposures such as hunting on a new infection with Pomona. Significantly associated with Pomona were “Work position” and two “Abattoirs” (GLM), and “Work position” (ABN). The odds of Pomona infection (OR, [95% CI]) was highest at stunning and hide removal (ABN 41.0, [6.9–1044.2]; GLM 57.0, [6.9–473.3]), followed by removal of intestines, bladder, and kidneys (ABN 30.7, [4.9–788.4]; GLM 33.8, [4.2–271.1]). Wearing a facemask, glasses or gloves (PPE) did not result as a protective factor in GLM or ABN. Conclusions/Significance The odds of Pomona infection was highest at stunning and hide removal. PPE did not show any indication of being protective in GLM or ABN. In ABN all relationships between variables are modelled; hence it has an advantage over GLM due to its capacity to capture the natural complexity of data more effectively. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Leptospirosis is one of the most extended zoonosis worldwide and humans become infected most commonly through contact with the urine of carrier animals, either directly or via contaminated water or soil. The aim in this study was to analyse the epidemiological behaviour of Leptospira spp., from domestic animals around the sites of human leptospirosis cases in Nicaragua, from 2007 through 2013. We report the results of a cross-sectional epidemiological study with a non-probability sampling of blood ( n = 3050) and urine ( n = 299) from Domestic Animals (DA) around the sites of human leptospirosis cases in Nicaragua. We analysed data obtained through Microscopic Agglutination Test (MAT), in-vitro culture, real time PCR and sequencing of lfb1 locus . Frequencies of 30.31% (95% CI: 28.66–31.95) and 15.38% (95% CI: 11.12–19.64) were obtained from serological test and from in-vitro culture, respectively. Although similar frequencies from serology test ( P ≥ 0.05) were found in DA species, in-vitro culture frequencies were significantly higher from bovine, equine and sheep ( P < 0.05) in comparison with swine and canine species. Ten serogroups of pathogenic Leptospira spp. were encountered, with the highest presence of Icterohaemorrhagiae serogroup 34.65% (95% CI: 29.35–39.94). We identified 7 samples homologous to L. interrogans species Pyrogenes serovar and 3 samples as L. noguchii Louisiana or Panama serovars by analysis of lfb1 sequences. We were able to establish a temporal and spatial correlation from DA and cumulative incidence of human cases. Therefore an effective epidemiological surveillance should be implemented with a specific control program toward DA in order to reduce human leptospirosis incidence. [ABSTRACT FROM AUTHOR]

Leptospirosis is a geographically widespread and globally underestimated zoonosis that affects humans and variety of animals. To identify trends and possible risk factors, joined medical and veterinary teams investigated epidemiology and epizootiology of leptospirosis in Croatia. Retrospective analysis of data obtained from referent diagnostic laboratories included a total of 1917 human and 123964 animal sera tested in the period from 2009 to 2014. We found high human leptospirosis average incidence rate of 1.53/100000 with clear predominance of male patients older than 40 years (sex ratio M/F:3.2; median age 51 ± 15.1 years). Statistical analysis revealed seasonal and annual variations of incidence in humans that were primarily associated with favourable weather conditions (temperature 10–19, 9 °C and precipitation above 100 mm/m 2 ). Majority of infections in humans were caused by serogroups Sejroe, Australis and Icterohaemorrhagiae. Notable variations in seroprevalence and changing trends in prevailing serogroups were recorded in most of the domestic animals and during the entire period of investigation. All of the observed findings underline leptospirosis as a significant human and veterinary public health threat and emphasize the importance of continuous multidisciplinary surveillance. We also argue that only input from both professions improves our overall knowledge on leptospirosis and leads to better and more efficient prevention and control strategies. [ABSTRACT FROM AUTHOR]

Leptospira spp., a zoonotic agent relevant for public health, occurs frequently in tropical regions. The aquatic environment represents a viable survival and transmission pathway. This study aimed to investigate the presence of anti- Leptospira spp. antibodies in Phrynops geoffroanus (Geoffroy’s side-necked turtle) serum samples using the microagglutination test (MAT), and Leptospira spp. in gastric and cloacal lavage samples using the polymerase chain reaction (PCR) technique. Antibodies against nine different Leptospira spp. serovars were detected in 45.45% (30/66) of the serum samples. Specific amplification of Leptospira spp. genomic material (331 bp) was observed in 16.67% (11/66) of the samples. In conclusion, these freshwater testudines host Leptospira spp. and eliminate them. This situation may represent a risk to public health, especially to people who use urban streams for fishing and recreational activities. Additionally, we described some Leptospira spp. serovars, not yet reported in testudines, detected here in P. geoffroanus . [ABSTRACT FROM AUTHOR]

Leptospirosis is an infectious, zoonotic disease of worldwide distribution, the cause of which is infection by pathogenic Leptospira. In Chile, dairy cattle are recognized a significant source in the maintenance and transmission of this infection, which causes economic losses and represents an infection threat to workers in the dairy industry. The infection is underestimated in cattle, due to the lack of clinical, pathognomonic signs, as well as the low efficiency of current diagnostic techniques. In this study, we developed antigen ELISA and dot blot assays, based on polyclonal antibodies, to detect pathogenic Leptospira in the urine samples of dairy cattle. The proposed tests showed an acceptable diagnostic accuracy, based on an analytical sensitivity of 1·104 Leptospira per mL for ELISA, and 3.2·103 for dot blot. These results corresponded with those obtained by qPCR, and the use of urine samples allowed us to propose new diagnostic alternatives for pathogenic Leptospira infection at a low cost, which can provide information on active infection status, which is a key element in control programs both at individual and herd level. [ABSTRACT FROM AUTHOR]

• Five-week-old female C3H/HeNJ mice intraperitoneally infected with low (103 cells) and high (106 cells) doses of Leptospira interrogans serovar Pomona did not develop acute lethal and survived up to 28 days after infection. • The high-dose infected C3H/HeNJ mice presented more pathological changes in the lungs and the kidneys including renal fibrosis and glomerular shrinkage than those infected with low-dose leptospires. • IFN-gamma, IL-6, and IgG2a in the high-dose infected C3H/HeNJ mice were induced at significantly higher levels than those in the low-dose infected mice. • C3H/HeNJ mice could be used as a sublethal model of leptospirosis. Leptospirosis is a worldwide zoonotic disease caused by pathogenic Leptospira spp. Leptospires can infect a variety of mammalian species. Golden Syrian hamsters are mostly used to study acute leptospirosis. However, the immunopathogenic mechanism is poorly understood because immunological reagents for hamsters are limited. This study aimed to establish C3H/HeNJ mice as an animal model for leptospirosis. Five-week-old C3H/HeNJ mice were infected with either low (103 cells) or high (106 cells) inoculum dose of Leptospira interrogans serovar Pomona. All mice were investigated for survival rate, leptospiral load and histopathology of target organs, antibody levels, and cytokine production (IFN-γ, IL-6 and IL-10) at day 28 post-infection. All infected mice survived and did not develop acute lethal infection. However, C3H/HeNJ mice infected with 106 cells of leptospires showed kidney colonization of leptospires and pathological changes in the lung and kidney including renal fibrosis. The glomerular size in PAS-D stained kidney tissues of C3H/HeNJ mice infected with 106 cells of leptospires was significantly reduced compared to that of mice infected with 103 cells of leptospires and non-infected mice. High-dose leptospires induced significantly greater levels of IFN-gamma and IL-6 than low-dose leptospires, but IL-10 level was not significantly different. Moreover, 106 leptospiral cells induced predominant IgG2a isotype suggesting Th1-like response. These results suggest that C3H/HeNJ mice may be used as a sublethal model of leptospirosis. [ABSTRACT FROM AUTHOR]

Leptospirosis is a zoonotic disease of global importance caused by pathogenic Leptospira species. Dogs can become asymptomatically infected, acting like reservoir hosts for pathogenic Leptospira, notably Leptospira interrogans serovar Canicola. Identification of such individuals and characterization of leptospires involved in chronic infections may unravel the role of dogs in the epidemiology of particular leptospiral strains. The aim of the present work was to describe the first Leptospira santarosai strain isolated from a dog. The dog was kept in a public shelter in São Paulo city, Brazil, and presented asymptomatic urinary shedding detected by PCR. Prospective evaluation was performed to fully characterize its chronic carrier state. The dog did not present anti- Leptospira titles or clinical/laboratorial abnormalities during the evaluations; nevertheless long-term urinary shedding was confirmed by PCR and leptospires were recovered from two occasions. The isolated strain was molecularly characterized by partial 16S rRNA and secY gene sequencing and MLST analysis. Serogroup identification was performed using polyclonal antibodies. The strain was identified as Leptospira santarosai , serogroup Sejroe. This is the first evidence in the literature of the isolation of L. santarosai in dogs. Our findings show that dogs can persistently harbor leptospires other than L. interrogans . [ABSTRACT FROM AUTHOR]

Leptospirosis is a worldwide spread zoonosis that can affect all groups of vertebrates, including reptiles. Because it has been little studied in snakes, this study focused on determining the occurrence of anti- Leptospira spp. antibodies in 64 Crotalus durissus collilineatus kept in captivity and on identifying the most common serovars in these animals, using the microscopic agglutination test. Of these, almost 90% were positive and there were reactions to the 22 serovars used in the study. The most common serovar in these snakes was Javanica, Andamana and Patoc. Most frequent titers were 25 and 50, although high titers (such as 1600) were also recorded, despite the absence of clinical symptoms. The possibility should be considered of captive snakes serving as a serious source of leptospiral infection in humans, which is why it is essential to study, prevent and control the disease in breeding centers and serpentariums. [ABSTRACT FROM AUTHOR]

Leptospirosis is an emerging disease, especially in countries with a tropical climate such as Malaysia. A dramatic increase in the number of cases has been reported over the last decade; however, information on the epidemiological trends of this disease is lacking. The objective of this study is to provide an epidemiological description of human leptospirosis cases over a 9-year period (2004–2012) and disease relationship with meteorological, geographical, and demographical information. A retrospective study was undertaken to describe the patterns of human leptospirosis cases and their association with intrinsic (sex, age, and ethnicity) and extrinsic (location, rainfall, and temperature) factors. Data was grouped according to age, sex, ethnicity, seasonality and geographical distribution, and analyzed using statistical tools to understand the influence of all the different factors on disease incidence. A total of 12,325 cases of leptospirosis were reported between 2004 and 2012 with an upward trend in disease incidence, with the highest in 2012. Three hundred thirty-eight deaths were reported with an overall case fatality rate of 2.74%, with higher incidence in males (9696; 78.7%) compared with female patients (2629; 21.3%), and overall male to female ratio of 3.69:1. Patients aged cohorts between 30–39 years old (16.22 per 100,000 population) had the highest disease incidence while the lowest incidence occurred between <1 to 9 years old (3.44 per 100,000 population). The average incidence was highest amongst Malays (10.97 per 100,000 population), followed by Indians (7.95 per 100,000 population). Stratification according to geographical distribution showed that the state of Malacca had the highest average disease incidence (11.12 per 100,000 population) followed by Pahang (10.08 per 100,000 population). The states of Terengganu, Kelantan, and Perak recorded similar rates of incidence (≈8.00 per 100,000 population), while Johor with the least number of reported cases (1.80 per 100,000 population). Positive relationships were recorded between the number of reported cases with the number of raining days per month and monthly average temperature ( p -value < 0.05). However, no significant association was noted between rainfall volume and number of reported Leptospirosis cases. This collaborative efforts between medical, academic and governmental institutions has enabled the construction of this comprehensive database that is essential to understand the disease trends in Malaysia and add insights into the prevention and control of this disease. [ABSTRACT FROM AUTHOR]

• The performance of different diagnostic techniques for leptospirosis in ewes raised in field semiarid conditions made it possible to identify carrier animals. • Genital route may be important for efficient transmission and without dependence on environmental factors in ewes from semiarid. • Titer 1:25 in serology was more efficient, indicating its use in ewes in field semiarid conditions. Leptospirosis is poorly studied in small ruminants raised in field semiarid conditions. In this study we compared serological, bacteriological and molecular diagnostic methods in ewes maintained in field Brazilian semiarid conditions. Blood, vaginal fluid and urine samples were collected from 60 Morada Nova ewes raised in a semi-intensive system 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 eight (13.33%) animals analyzed by MAT at reciprocal titer 25 (cut-off 25), while Leptospira sp. DNA was detected in urine or vaginal fluid of 56 animals (93.33%). There was growth of leptospires in 10 urine cultures and in 11 vaginal fluid cultures, however, two of urine (2/60–3.33%) and eight cultures of vaginal fluid (8/60–13.33%) were confirmed by PCR. Two samples of vaginal fluid (one of each animal) were submitted to sequencing demonstrating 99% similarity with L. santarosai and L. interrogans. The highest MAT sensitivities were obtained with reciprocal titer 25 (cut-off 25) compared to 50 and 100. The performance of different diagnostic techniques for leptospirosis in ewes raised in field semiarid conditions allowed a better evaluation of the herd, as well as made it possible to identify carrier animals. Genital route may be important for efficient transmission and without dependence on environmental factors in ewes from semiarid, as well as it's highlighted that titer 1:25 in serology was more efficient, indicating its use in ewes in field semiarid conditions. [ABSTRACT FROM AUTHOR]

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 (MAT 21 ). Additionally, 12 local strains (MAT 33 ) were included as antigens. PCR was performed with the urine samples and it was positive on 71 out of 222 samples (31.9%). MAT 21 identified as seroreactive 173 (55.1%) out of the 314 animals studied, with Sejroe the most common (38.1%). In MAT 33 , 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. [ABSTRACT FROM AUTHOR]

Toll-like receptor 2 (TLR2), a member of the Toll-like receptor family, plays an important role in the recognition of and subsequent immune response activation against leptospirosis in humans. The genetic polymorphism in TLR2 of an arginine to glutamine substitution at residue 753 (Arg753Gln) has been associated with a negative influence on TLR2 function, which may, in turn, determine the innate host response to Leptospira spp. This bacterium signals through TLR2/TLR1 heterodimers in human cells. The aim of the present study was to investigate the Arg753Gln single-nucleotide polymorphism (SNP) of the TLR2 gene, and the isoleucine to serine transversion at position 602 (Ile602Ser) of the TLR1 gene (previously associated with Lyme disease), in leptospirosis patients compared to healthy controls, carrying out a retrospective case/control study. The TLR2 polymorphism adenine (A) allele was observed in 7.3% of leptospirosis patients but was not found in the control group, whereas the guanine (G) allele of the TLR1 polymorphism was found in 63.6% of patients and 41.6% of controls. Susceptibility to leptospirosis disease was increased 10.57-fold for carriers of the TLR2 G/A genotype ( P = 0.0493) and 3.85-fold for carriers of the TLR1 G/G genotype ( P = 0.0428). Furthermore, the risk of developing hepatic insufficiency and jaundice was increased 18.86- and 27.60-fold for TLR2 G/A carriers, respectively. Similarly, the risk of developing jaundice was increased 12.67-fold for TLR1 G allele carriers (G/G and T/G genotypes). In conclusion, the present data suggest that the TLR2 Arg753Gln and TLR1 Ile602Ser SNPs influence the risk of developing leptospirosis and its severity. [ABSTRACT FROM AUTHOR]

Evolving land use practices have led to an increase in interactions at the human/wildlife interface. The presence and poor knowledge of zoonotic pathogens in India's wildlife and the occurrence of enormous human populations interfacing with, and critically linked to, forest ecosystems warrant attention. Factors such as diverse migratory bird populations, climate change, expanding human population and shrinking wildlife habitats play a significant role in the emergence and re-emergence of zoonotic pathogens from India's wildlife. The introduction of a novel Kyasanur forest disease virus (family flaviviridae) into human populations in 1957 and subsequent occurrence of seasonal outbreaks illustrate the key role that India's wild animals play in the emergence and reemergence of zoonotic pathogens. Other high priority zoonotic diseases of wildlife origin which could affect both livestock and humans include influenza, Nipah, Japanese encephalitis, rabies, plague, leptospirosis, anthrax and leishmaniasis. Continuous monitoring of India's extensively diverse and dispersed wildlife is challenging, but their use as indicators should facilitate efficient and rapid disease-outbreak response across the region and occasionally the globe. Defining and prioritizing research on zoonotic pathogens in wildlife are essential, particularly in a multidisciplinary one-world one-health approach which includes human and veterinary medical studies at the wildlife-livestock-human interfaces. This review indicates that wild animals play an important role in the emergence and re-emergence of zoonotic pathogens and provides brief summaries of the zoonotic diseases that have occurred in wild animals in India. [ABSTRACT FROM AUTHOR]

Highlights: [•] Leptospira carriage rate and circulating serovars investigated in small mammals. [•] Very high carriage rate detected (29.9%). [•] Three genomic species and five serovars detected. [•] Serogroups in small mammals correlate with serogroups in humans in that area. [ABSTRACT FROM AUTHOR]

Abstract: In 2009, a survey was conducted in Reunion Island to determine the renal leptospiral load in black rats trapped in the field. The concentration of leptospires in kidney tissue was calculated using qPCR. Our results showed high inter-individual variations of renal bacterial load in naturally infected black rats (mean=8.27±4.72 log-genome copies per mg kidney tissue). The objective of this study was to model the renal leptospiral load in 50 naturally infected black rats as a function of sex, age, and weight. Statistical analysis by sex showed that, in naturally infected males, the renal leptospiral load was correlated with weight (p-value=0.032). Moreover, our model showed that weight and sex were significant explanatory variables for the renal leptospiral load in naturally infected young black rats (R 2 =0.953). Laboratory experimentation could not replicate naturally acquired infection, but field studies also present many limitations. Our study is the first attempt to explain individual variations in the renal leptospiral load in naturally infected reservoir animals but further research is needed. [Copyright &y& Elsevier]

Abstract: Leptospirosis is a zoonotic disease that occurs all over the world, caused by bacteria of the genus Leptospira. Marsupial and didelphidae families are considered susceptible to infection caused by a wide range of Leptospira serovars for which they serve as reservoirs. Thirty-three free-living white-eared opossums (Didelphis albiventris) were captured in Southern Brazil and bodily fluids were collected. From the urine samples it was possible to obtain an isolate identified as Leptospira borgpetersenii by rpoB gene sequencing and belonging to serovar Castellonis by Multilocus Variable-Number Tandem-Repeat Analysis. This is the first report of the isolation of Leptospira spp. from the white-eared opossum in Brazil. In addition, the new strain was also virulent in the hamster model of lethal leptospirosis. The microscopic agglutination test (MAT) was used for detecting the presence of antibodies against Leptospira spp. in white-eared opossum, human, cattle and canine sera using a panel of 59 Leptospira strains that included the new isolate. The inclusion of the new strain in the MAT battery increased the MAT sensitivity for canine sera. These findings suggest that the white-eared opossum is an important reservoir of pathogenic Leptospira spp. [Copyright &y& Elsevier]

Abstract: Climate change, world population growth, and poverty have led to an increase in the incidence of leptospirosis. Leptospirosis is caused by pathogenic spirochaete bacteria that belong to the genus Leptospira. The bacteria are maintained in the renal tubules of the reservoir hosts (typically a rodent), then shed into the environment via the urine. Water is key for environmental survival and transmission, as leptospires can survive for several weeks in a moist environment. Therefore, environmental epidemiological studies are needed to study the contamination of environmental water sources. However, few such studies have been performed using cultivation of the isolates and PCR assays. But, leptospira cultivation can be easily contaminated by other organisms and takes usually several weeks. Moreover, PCR is a complex and costly analysis for the underdeveloped countries that have the highest incidence of leptospirosis. In this study, we describe two modifications of a fluorescence microscopy assay based on immuno-magnetic separation (IMS) to detect leptospires in environmental water samples that mainly differ in fluorescent dye staining. The first type uses acridine orange fluorescent dye staining combined with multiplexed IMS for sample screening. The detection limit ranged from 102 to 103 organisms per mL and largely depended on the capture efficiency (CE) of the immuno-magnetic particles. The second type uses serogroup-specific immuno-particles and direct fluorescence antibody staining (DFA) to detect leptospires; the detection limit of this second assay was approximately 101 cells per mL. Both assay types were applied to natural and experimentally infected water samples, which were also analysed with DFM and real-time PCR. Our data show that the fluorescent microscopy immunoassay successfully identified experimental leptospire contamination and was as sensitive as PCR. This modified immune-fluorescence assay may therefore enable epidemiological studies of leptospirosis. [Copyright &y& Elsevier]

Abstract: The aim of the study was to investigate the effect of infection with Leptospira spp. in horses on activities of platelet-activating factor acetylhydrolase (PAF-AH) and paraoxonase-1 (PON1) considering the anti-inflammatory/anti-oxidative properties of both enzymes. A total of 63 sport and leisure horses’ sera were chosen among those routinely serologically examined on leptospirosis during the year 2009 in Croatia. Sera were divided into three groups according to the estimated level of antibody titre against Leptospira spp; group 1: sera serologically negative to leptospirosis (antibody titre<50) (n =21); group 2: sera with residual or anamnestic antibody titre (50–200) (n =23); group 3: sera with high antibody titre (≥1600) indicating recent Leptospira infection (n =19). Serum PAF-AH and PON1 activity was not significantly different between investigated groups (p >0.05). There were no statistical differences in lipid status parameters (p >0.05) among study groups as well. However, significant positive correlations (p <0.05) of PAF-AH and PON1 with total cholesterol and HDL–C were found. Results may indicate low levels of systemic inflammatory response and oxidative stress in horses with subclinical leptospirosis. Further studies with clinically manifested disease are needed to elucidate the potential role of PAF-AH and PON1 as prognostic markers of the leptospirosis outcome. [Copyright &y& Elsevier]

Abstract: The frequency of massive pulmonary hemorrhages seems to be increasing in different geographic areas; however, there is no clear explanation for this trend. Although data on the pathogenesis of such complications are scarce, recent research indicates a potential role of autoimmunity and/or multifactorial mechanisms. However, much information is already available on the disturbance of hemostasis and blood vessels in leptospirosis-related literature, even if some contradictory concepts coexist. The purpose of this review is to integrate both new and classical information from human and animal studies on severe pulmonary forms of leptospirosis and disorders of hemostasis and blood vessels. We propose that the involvement of blood vessels in leptospirosis must be understood as a sepsis-like, diffuse process of endothelial activation/damage rather than as a classical systemic vasculitis. Pulmonary hemorrhages are most likely multifactorial and there has recently been evidence against the role of autoimmunity; however, further investigation of strain variations, exposure to hydrocarbons and association with renal dysfunction is required. Thrombocytopenia is a consistent feature of leptospirosis but it is not clear whether it is attributable to sepsis-related mechanisms. In addition, further investigation is required to define whether platelet function is activated or inhibited during severe leptospirosis. [Copyright &y& Elsevier]

Leptospiral infection is widespread in wildlife. In this context, wild ecosystems in tropical countries hold a vast biodiversity, including several species that may act as potential reservoirs of leptospires. The Pantanal biome presents highly favorable environmental conditions for the occurrence of leptospirosis, such as high temperatures, constant flooding, and high biodiversity. The purpose of this study was to detect wild animals as carriers of Leptospira sp. using direct methods (PCR and culture) in the Pantanal biome, Brazil. A total of 35 animals were studied, namely Cerdocyon thous, Nasua nasua , Ozotoceros bezoarticus, and Sus scrofa species. Blood for serology (MAT) and urine for bacteriological culturing and PCR was sampled. The most prevalent serogroups were Javanica and Djasiman. Additionally, 40.6% of these animals presented PCR positive reactions. Seroreactivity associated with the high frequency of leptospiral carriers among the different studied species suggests a high level of exposure of the studied animals to pathogenic Leptospira strains. Our results are still limited and the actual role of the studied animals in the epidemiology of leptospirosis in the Pantanal region remains to be elucidated. [ABSTRACT FROM AUTHOR]

Abstract: A retrospective study to describe the epidemiology of human leptospirosis in Trinidad and Tobago during 1996–2007 was conducted. All confirmed cases of leptospirosis was analyzed according to age, sex, seasonality and geographic distribution. A total of 278 cases were recorded, with an average annual incidence rate of 1.84 per 100,000 population. Seventy-five percent of the cases occurred during the wet season, with the highest number of cases recorded in November. A positive correlation was found between number of cases and rainfall. Males constituted 80% of all cases, and the overall male:female ratio was 4.6:1. The total case fatality rate was 5.8%, with deaths among males four times more common than in females. Clinical leptospirosis was greatest in the 10–19 age group and lowest in the 0–9 age group. The total prevalence was 22 per 100,000 population, with the highest prevalence 41 per 100,000 recorded in the regional corporation of Sangre Grande and the lowest (6 per 100,000) in the city of Port of Spain. The lack of important information and active surveillance showed that the level of awareness of the disease is low in the country. The disease is still under-reported, and is considered to be of significant public health importance. [Copyright &y& Elsevier]