Polymerase Chain Reaction and Dot-Blot Hybridization for Leptospira Detection in Water Samples.

The dot-blot is a simple, fast, sensitive, and versatile technique that enables the identification of minimal quantities of DNA specifically targeted by probe hybridization in the presence of carrier DNA. It is based on the transfer of a known amount of DNA onto an inert solid support, such as a nylon membrane, utilizing the dot-blot apparatus and without electrophoretic separation. Nylon membranes have the advantage of high nucleic acid binding capacity (400 µg/cm2), high strength, and are positively or neutrally charged. The probe used is a highly specific ssDNA fragment of 18 to 20 bases long labeled with digoxigenin (DIG). The probe will conjugate with the Leptospira DNA. Once the probe has hybridized with the target DNA, it is detected by an anti-digoxigenin antibody, allowing its easy detection through its emissions revealed in an X-ray film. The dots with an emission will correspond to the DNA fragments of interest. This method employs the non-isotopic labeling of the probe, which may have a very long half-life. The drawback of this standard immuno-label is a lower sensitivity than isotopic probes. Nevertheless, it is mitigated by coupling polymerase chain reaction (PCR) and dot-blot assays. This approach enables the enrichment of the target sequence and its detection. Additionally, it may be used as a quantitative application when compared against a serial dilution of a well-known standard. A dot-blot application to detect Leptospira from the three main clades in water samples is presented here. This methodology can be applied to large amounts of water once they have been concentrated by centrifugation to provide evidence of the presence of Leptospiral DNA. This is a valuable and satisfactory tool for general screening purposes, and may be used for other non-culturable bacteria that may be present in water, enhancing the comprehension of the ecosystem.

Risk practices and awareness of leptospirosis amongst residents of Zaria, Nigeria.

This study evaluated the level of risk practices and awareness of leptospirosis among residents of Zaria, Nigeria. A pre-tested questionnaires were administered via face-to-face interview to 100 residents. The data was analyzed using chi-square and multivariate analysis to identify risk factors for leptospirosis. The demography showed that the majority of the respondents were male, aged 21-40 years, and majorly crop farmers. The risk factors identified showed that males were 4.14 times more likely to be affected by leptospirosis (OR 4.14, 95% CI [1.93-5.37], p = 0.02) and the source of animal's water was 5.56 times more likely to be contaminated by Leptospira spp. (OR 4.14, 95% CI [2.88-8.03], p = 0.01) and these relationships were significant. The majority of respondents were not aware of the disease (OR 1.87, 95% CI [1.22-4.57], p = 0.01) with 78% of the respondents not sure of which of the animal species leptospirosis affected (OR 1.67, 95% CI [1.07-2.62], p = 0.02). This study has demonstrated the existence of risk behaviors, and paucity of knowledge about leptospirosis in the study area. It is therefore recommended to organize an enlightenment program and the need for protective clothing for individuals occupationally at risk of infection by Leptospira spp.

Molecular and serological prevalence of Leptospira spp. among slaughtered cattle and associated risk factors in the Bahr El Ghazal region of South Sudan.

INTRODUCTION: Leptospirosis is a neglected emerging and zoonotic disease reported worldwide. This study sought to determine the molecular and serological prevalence of Leptospira spp. and the associated risk factors in slaughtered cattle from the Bahr El Ghazal region of South Sudan. MATERIALS AND METHODS: Between January 16th and February 25th, 2023, blood and urine samples were collected from 402 cattle at the Lokoloko Municipal Slaughterhouse in Western Bahr El-Ghazal State. Serum samples were tested using the microscopic agglutination test (MAT), with a panel of 12 serovars (sv) from 12 serogroups (sg) and 4 species (spp) of Leptospira spp. These serovars had been previously identified in Sudan and the East African region. Simultaneously, 400 corresponding urine samples were screened using qualitative real-time polymerase chain reaction (PCR) to detect the shedding of Leptospira spp. in urine. To identify the associated risk factors, the age, sex, breed and body condition score of each sampled cattle was noted at the time of sampling and subsequently analysed using logistic regression models. RESULTS: Among the 402 serum samples screened, a substantial 81.8% (329/402, 95% CI 77.9-85.3) displayed seropositivity for Leptospira spp. with a MAT titre ≥ 100. The prevalence of urine shedding determined by PCR was 6% (23/400, 95% CI 3.8-8.4), while probable recent leptospirosis with a MAT ≥ 1:800 was observed in 33.1% (133/402, 95% CI 28.6-37.8) of the cattle. Multiple reactions were detected in 34.8% (140/402, 95% CI 30.6-39.5) serum samples. The seropositivity was against L. borgpetersenii sg. Tarassovi (78.6%; 316/402, 95% CI 74.4-82.3), followed by L. borgpetersenii sg. Ballum at 20.4% (82/402, 95% CI, 16.7-24.4%), L. kirschneri sg. Autumnalis At 8.7% (35/402, 95% CI 5.7-11.7), L. interrogans sg. of Pomona at 7.0% (28/402, 95% CI 4.5-9.5), and L. interrogans sg. Hebdomadis was 5.0% (20/402, 95% CI 2.8-7.2). Several risk factors are associated with seropositivity. Older animals (≥ 2 years) had 2.0 times greater odds (95% CI 1.14-3.5) of being seropositive than younger animals (< 2 years), P-value = 0.016. Female animals demonstrated 2.1 times greater odds (95% CI 1.2-3.6) of seropositivity than males did (P-value = 0.008). Additionally, Felata/Mbororo cattle exhibited 2.4 times greater odds (95% CI 1.3-4.5) of being seropositive than did local Nilotic cattle (P-value = 0.005). The agreement between the MAT and PCR results was poor, as indicated by a kappa statistic value of 0.001 and a P-value of 0.913. But there was a moderate agreement between MAT high titres ≥ 800 and PCR positivity with a kappa statistic value = 0.501 and a P-value < 0.001. CONCLUSION: In addition to the high seroprevalence, Leptospira spp. were found in the urine of slaughtered cattle, suggesting that leptospirosis is endemic to the study area. This finding underscores the significance of cattle as potential sources of infection for slaughterhouse workers, the general public, and other animal species. To address this issue effectively in the Bahr El Ghazal Region and South Sudan, a comprehensive strategy involving a multidisciplinary approach is essential to minimize disease among animals, hence reducing potential zoonotic risks to humans.

Prevalence and transmission of the most relevant zoonotic and vector-borne pathogens in the Yucatan peninsula: A review.

BACKGROUND: Habitat modification and land use changes impact ecological interactions and alter the relationships between humans and nature. Mexico has experienced significant landscape modifications at the local and regional scales, with negative effects on forest cover and biological biodiversity, especially in the Yucatan peninsula in southeastern Mexico. Given the close relationship between landscape modification and the transmission of zoonotic and vector-borne diseases, it is essential to develop criteria for identifying priority zoonoses in the south of the country. METHODOLOGY/PRINCIPAL FINDINGS: We reviewed 165 published studies on zoonotic and vector-borne diseases in the region (2015-2024). We identified the most frequent vectors, reservoirs, and hosts, the most prevalent infections, and the factors associated with transmission risk and the anthropogenic landscape modification in urban, rural, ecotone, and sylvatic habitats. The most relevant pathogens of zoonotic risk included Trypanosoma cruzi, arboviruses, Leishmania, Rickettsia, Leptospira, and Toxoplasma gondii. Trypanosoma cruzi was the vector-borne agent with the largest number of infected vertebrate species across habitats, while Leishmania and arboviruses were the ones that affected the greatest number of people. Dogs, cats, backyard animals, and their hematophagous ectoparasites are the most likely species maintaining the transmission cycles in human settlements, while rodents, opossums, bats, and other synanthropic animals facilitate connection and transmission cycles between forested habitats with human-modified landscapes. Pathogens displayed different prevalences between the landscapes, T. cruzi, arbovirus, and Leptospira infections were the most prevalent in urban and rural settlements, whereas Leishmania and Rickettsia had similar prevalence across habitats, likely due to the diversity and abundance of the infected vectors involved. The prevalence of T. gondii and Leptospira spp. may reflect poor hygiene conditions. Additionally, results suggest that prevalence of zoonotic and vector-borne diseases is higher in deforested areas and agricultural aggregates, and in sites with precarious health and infrastructure services. CONCLUSIONS: Some hosts, vectors, and transmission trends of zoonotic and vector-borne diseases in the YP are well known but others remain poorly recognized. It is imperative to reinforce practices aimed at increasing the knowledge, monitoring, prevention, and control of these diseases at the regional level. We also emphasize the need to perform studies on a larger spatio-temporal scale under the socio-ecosystem perspective, to better elucidate the interactions between pathogens, hosts, vectors, environment, and sociocultural and economic aspects in this and many other tropical regions.

Quantum dots as a fluorescent labeling tool for live-cell imaging of Leptospira.

Leptospirosis is a global public health problem caused by Gram-negative pathogenic bacteria belonging to the genus Leptospira. The disease is transmitted through the urine of infected animals, which contaminates water and soil, leading to the infection of other animals and humans. Currently, several approaches exist to detect these bacteria; however, a new sensitive method for the live-cell imaging of Leptospira is required. In this study, we report the green synthesis of cadmium telluride quantum dots (CdTe QDs) which are unique fluorescent nanocrystals with a high fluorescence quantum yield capable of modifying cell surfaces and are biocompatible with cells. The fabrication of QDs with concanavalin A (ConA), a carbohydrate-binding lectin and known biological probe for Gram-negative bacteria, produced ConA-QDs which can effectively bind on Leptospira and exhibit strong fluorescence under simple fluorescence microscopy, allowing the live-cell imaging of the bacteria. Overall, we performed the simple synthesis of ConA-QDs and demonstrated their potential use as versatile fluorescent probes for the live-cell imaging of Leptospira. This technique could be further applied to track leptospiral cells and study the infection mechanism, contributing to a more thorough understanding of leptospirosis and how to control it in the future.

Chimeric lipoproteins for leptospirosis vaccine: immunogenicity and protective potential.

Leptospirosis, a neglected zoonotic disease, is caused by pathogenic spirochetes belonging to the genus Leptospira and has one of the highest morbidity and mortality rates worldwide. Vaccination stands out as one of the most effective preventive measures for susceptible populations. Within the outer membrane of Leptospira spp., we find the LIC12287, LIC11711, and LIC13259 lipoproteins. These are of interest due to their surface location and potential immunogenicity. Thorough examination revealed the conservation of these proteins among pathogenic Leptospira spp.; we mapped the distribution of T- and B-cell epitopes along their sequences and assessed the 3D structures of each protein. This information aided in selecting immunodominant regions for the development of a chimeric protein. Through gene synthesis, we successfully constructed a chimeric protein, which was subsequently expressed, purified, and characterized. Hamsters were immunized with the chimeric lipoprotein, formulated with adjuvants aluminum hydroxide, EMULSIGEN®-D, Sigma Adjuvant System®, and Montanide™ ISA206VG. Another group was vaccinated with an inactivated Escherichia coli bacterin expressing the chimeric protein. Following vaccination, hamsters were challenged with a virulent L. interrogans strain. Our evaluation of the humoral immune response revealed the production of IgG antibodies, detectable 28 days after the second dose, in contrast to pre-immune samples and control groups. This demonstrates the potential of the chimeric protein to elicit a robust humoral immune response; however, no protection against challenge was achieved. While this study provides valuable insights into the subject, further research is warranted to identify protective antigens that could be utilized in the development of a leptospirosis vaccine. KEY POINTS: • Several T- and B-cell epitopes were identified in all the three proteins. • Four different adjuvants were used in vaccine formulations. • Immunization stimulated significant levels of IgG2/3 in vaccinated animals.

Surveillance of human leptospirosis infections in Ukraine between 2018 and 2023.

Leptospirosis is a bacterial disease that affects both humans and animals worldwide. Currently, a positional war is ongoing in Ukraine, and the military is encountering a significant number of rodents in trenches and dugouts, which are known reservoirs for Leptospira, the causative agent of leptospirosis-a potentially dangerous infectious disease with a high mortality rate. The civilian population is also at potential risk of leptospirosis. The destruction of the Kakhovka Dam on June 6, 2023, has led to widespread devastation and human suffering. In the short term, there is a significant risk of rodent-borne diseases such as leptospirosis. We utilized data from the Ukrainian Centre for Disease Prevention Control and observed a substantial increase in prevalence in 2023. The notification rate in Ukraine in 2023 was 1.06 per 100,000 persons, which is higher than that of other countries in the European Union. Particular attention is being given to Zakarpattia Oblast, located on the western border of Ukraine, which shares boundaries with Romania, Hungary, Poland, and Slovakia, with an extremely high incidence rate of 12.08 per 100,000 persons. Based on these findings, we recommend education and awareness campaigns, vaccination, personal protective measures, and improved surveillance to address the increasing incidence of leptospirosis in Ukraine.

Detection of Leptospira kirschneri in a short-beaked common dolphin (Delphinus delphis delphis) stranded off the coast of southern California, USA.

BACKGROUND: Pathogenic Leptospira species are globally important zoonotic pathogens capable of infecting a wide range of host species. In marine mammals, reports of Leptospira have predominantly been in pinnipeds, with isolated reports of infections in cetaceans. CASE PRESENTATION: On 28 June 2021, a 150.5 cm long female, short-beaked common dolphin (Delphinus delphis delphis) stranded alive on the coast of southern California and subsequently died. Gross necropsy revealed multifocal cortical pallor within the reniculi of the kidney, and lymphoplasmacytic tubulointerstitial nephritis was observed histologically. Immunohistochemistry confirmed Leptospira infection, and PCR followed by lfb1 gene amplicon sequencing suggested that the infecting organism was L.kirschneri. Leptospira DNA capture and enrichment allowed for whole-genome sequencing to be conducted. Phylogenetic analyses confirmed the causative agent was a previously undescribed, divergent lineage of L.kirschneri. CONCLUSIONS: We report the first detection of pathogenic Leptospira in a short-beaked common dolphin, and the first detection in any cetacean in the northeastern Pacific Ocean. Renal lesions were consistent with leptospirosis in other host species, including marine mammals, and were the most significant lesions detected overall, suggesting leptospirosis as the likely cause of death. We identified the cause of the infection as L.kirschneri, a species detected only once before in a marine mammal - a northern elephant seal (Mirounga angustirostris) of the northeastern Pacific. These findings raise questions about the mechanism of transmission, given the obligate marine lifestyle of cetaceans (in contrast to pinnipeds, which spend time on land) and the commonly accepted view that Leptospira are quickly killed by salt water. They also raise important questions regarding the source of infection, and whether it arose from transmission among marine mammals or from terrestrial-to-marine spillover. Moving forward, surveillance and sampling must be expanded to better understand the extent to which Leptospira infections occur in the marine ecosystem and possible epidemiological linkages between and among marine and terrestrial host species. Generating Leptospira genomes from different host species will yield crucial information about possible transmission links, and our study highlights the power of new techniques such as DNA enrichment to illuminate the complex ecology of this important zoonotic pathogen.

Leptospira spp. and Rickettsia spp. as pathogens with zoonotic potential causing acute undifferentiated febrile illness in a central-eastern region of Peru.

OBJETIVE: this study was to determine the relationship between acute febrile illness and bacterial pathogens with zoonotic potential that cause emerging and re-emerging diseases in a central-eastern region of Peru. RESULTS: Out of the 279 samples analyzed, 23 (8.2%) tested positive for infection by Rickettsia spp., while a total of 15 (5.4%) tested positive for Leptospira spp. Women had a higher frequency of infection by Rickettsia spp., with 13 cases (53.3%), while men had a higher frequency of infection by Leptospira spp., with 10 cases (66.7%). The most frequently reported general symptom was headache, with 100.0% (n = 23) of patients with Rickettsia (+) and 86.7% (n = 13) of patients with Leptospira (+) experiencing it. Arthralgia was the second most frequent symptom, reported by 95.6% (n = 22) and 60% (n = 9) of patients with Rickettsia (+) and Leptospira (+), respectively. Myalgia was reported by 91.3% (n = 21) and 66.7% (n = 10) of patients with Rickettsia (+) and Leptospira (+), respectively. Retroocular pain, low back pain, and skin rash were also present, but less frequently. Among the positives, no manifestation of bleeding was recorded, although only one positive case for Leptospira spp. presented a decrease in the number of platelets.

Spatiotemporal assessment of pathogenic Leptospira in subtropical coastal watersheds.

The World Health Organization classifies leptospirosis as a significant public health concern, predominantly affecting impoverished and unsanitary regions. By using the Pensacola Bay System as a case study, this study examines the underappreciated susceptibility of developed subtropical coastal ecosystems such as the Pensacola Bay System to neglected zoonotic pathogens such as Leptospira. We analyzed 132 water samples collected over 12 months from 44 distinct locations with high levels of Escherichia coli (>410 most probable number/100 mL). Fecal indicator bacteria (FIB) concentrations were assessed using IDEXX Colilert-18 and Enterolert-18, and an analysis of water physiochemical characteristics and rainfall intensity was conducted. The LipL32 gene was used as a quantitative polymerase chain reaction (qPCR) indicator to identify the distribution of Leptospira interrogans. The results revealed 12 instances of the presence of L. interrogans at sites with high FIB over various land cover and aquatic ecosystem types. Independent of specific rainfall events, a seasonal relationship between precipitation and elevated rates of fecal bacteria and leptospirosis was found. These findings highlight qPCR's utility in identifying pathogens in aquatic environments and the widespread conditions where it can be found in natural and developed areas.

THREE CASES OF CLINICAL LEPTOSPIROSIS IN PATAGONIAN MARAS (DOLICHOTIS PATAGONUM).

Rodents are typically viewed as asymptomatic reservoirs for leptospirosis infection, as clinical disease in rodents is rarely described. This report includes three separate cases of leptospirosis in Patagonian maras (Dolichotis patagonum) over a 3-yr period in multiple locations within a single zoo. All three cases presented with varying clinical signs including lethargy, conjunctival hyperemia, hyperbilirubinemia, and presumed renal azotemia. Infection with Leptospira spp. was diagnosed antemortem by PCR on whole blood (n = 1, Case 1) or urine (n = 2, Cases 2 and 3). Leptospira antibody titers measured by serum microagglutination testing (n = 3) were elevated or increased in all three animals over a 1-3-wk period for Leptospira serovars Bratislava and Hardjo (Case 1) and Grippotyphosa (Case 2 and 3). Two of the three animals responded to treatment with penicillin and doxycycline and supportive care, whereas one animal did not respond to treatment. Postmortem findings in this individual included conjunctivitis, chemosis, dehydration, icterus, tricavitary serosanguinous effusions, necrotizing hepatitis, diffuse pulmonary congestion, and edema. Immunohistochemical examination identified scattered Leptospira organisms within hepatocytes and renal tubular epithelial cells. A wild raccoon (Procyon lotor) at the institution tested positive by PCR on kidney tissue for the same Leptospira spp. serovar and was the suspected source of infection. This case series highlights the clinical importance of leptospirosis as a differential for Patagonian maras presenting with lethargy, ocular signs, acute hepatic disease, and azotemia.

Seroprevalence of Leptospira antibodies in dogs and cats from Santa Fe, a city in East-Central Argentina endemic for leptospirosis.

This study examines household pets as potential epidemiological links between environments contaminated with pathogenic leptospires and humans in Santa Fe, Argentina. The aims of our study were: (a) to characterize the habits and exposure to environmental sources of leptospirosis in the population of dogs and cats attending to municipal spay and neutering campaigns in Santa Fe, Argentina, (b) to assess the seroprevalence of anti-Leptospira antibodies in asymptomatic dogs and cats, (c) to evaluate factors that could increase seropositivity, and (d) to identify spatial clusters of seropositive dogs and cats in the capital city of Santa Fe. From May to November 2022, a cross-sectional serosurvey was conducted during municipal spaying/neutering campaigns. Eligible household dogs and cats were over 6 months old, apparently healthy, and not vaccinated against leptospirosis in the past 6 months. We used microagglutination test (MAT) to assess anti-Leptospira antibodies using a panel of 10 reference strains. We used generalized linear mixed effects models (GLMM) to examine individual and census tract-level risk factors for seropositivity, and local Moran's I statistic for spatial clusters. Results showed higher leptospiral antibody prevalence in dogs (18.2 %) than cats (3.6 %, p = 0.002). Dogs with street access had higher likelihood of being seropositive (OR: 3.8, 95 % CI: 1.2; 11.9), and areas with chronic poverty showed an elevated risk of presenting seropositive animals (RR: 4.0, 95 % CI: 1.1; 14.4). Spatial analysis didn't reveal significant seropositivity clusters among census tracts. These findings shed light on widespread Leptospira seropositivity in pets in this endemic region. Understanding seroprevalence and risk factors can guide public and veterinary health strategies, emphasizing increased leptospirosis vaccination for dogs in vulnerable areas and promoting responsible pet care.

Estimation of economic losses due to leptospirosis in dairy cattle.

There are few studies that effectively quantify the economic losses resulting from problems caused by leptospirosis in naturally infected dairy cattle. Given this gap, the objective of this study was to propose and apply a method to quantify the economic losses resulting from productive and reproductive problems in a commercial dairy herd naturally infected by Leptospira spp. For this study, the zootechnical and economic indicators at a property with Jersey cattle were analyzed during the period from 2014 to 2017. The leptospirosis outbreak occurred in 2014, and the therapeutic approach was carried out between 2015 and 2017, with the latter considered the year of control of the outbreak. The adopted integrated control strategy consisted of dividing the herd according to the serological results obtained through the microscopic agglutination test, the treatment of reagents with streptomycin, and vaccination against leptospirosis of non-reagent heifers and cows. The method used to evaluate the economic indicators of the property was the calculation of the gross margin by taking into account the implicit and explicit cost parameters associated with the manifestation of leptospirosis. The prevalence rate of leptospirosis decreased from 49.4 % in 2015 to 21.6 % in 2017. There was a reduction in the abortion rate (from 40.00 % in 2014 to 9.00 % in 2017), in the stillborn rate (from 2.63 % in 2014 to 1.69 % in 2017) and an increase in the calving rate (from 65.00 % in 2014 to 86.00 % in 2017). In addition, there were increases in the number of lactating cows (from 38 in 2014-57 in 2017) and the mean times of lactation duration, which increased from 275 days in 2014-295 days in 2017. As a result, the average annual production of milk increased from 164,655 liters in 2014-248,521 liters in 2017. In 2014, when treatment hadn't yet started, the gross margin per liter of milk sold, considering implicit and explicit costs, was US$0.00. In 2015 and 2016, US$0.27 and US$0.30 were obtained, respectively, for this variable. In 2017, with the disease under control on the property, the gross margin per liter of milk reached US$0.36. The gross margin per liter of milk sold was higher in the period when the disease was controlled, showing losses of up to 84 % of the gross margin during the outbreak. Immediate treatment of positive cows and preventive measures had a significant impact on improving the productive and economic efficiency of the property.

Antibodies against Leptospira spp. in Free-living and Captive Broad-snouted Caiman (Caiman latirostris) and Free-living Yacare Caiman (Caiman yacare) in Brazil.

We evaluated antibodies against Leptospira spp. in both free-living and captive Caiman latirostris from Atlantic Forest, and free-living Caiman yacare from Pantanal, Brazil, by using a microscopic agglutination test. Overall seropositivity was 17%, with rates of 36% in captive C. latirostris (n=4/11) and 18% in free-living C. yacare (n=4/22).

Molecular and serological diagnosis of multiple bacterial zoonoses in febrile outpatients in Garissa County, north-eastern Kenya.

Bacterial zoonoses are diseases caused by bacterial pathogens that can be naturally transmitted between humans and vertebrate animals. They are important causes of non-malarial fevers in Kenya, yet their epidemiology remains unclear. We investigated brucellosis, Q-fever and leptospirosis in the venous blood of 216 malaria-negative febrile patients recruited in two health centres (98 from Ijara and 118 from Sangailu health centres) in Garissa County in north-eastern Kenya. We determined exposure to the three zoonoses using serological (Rose Bengal test for Brucella spp., ELISA for C. burnetti and microscopic agglutination test for Leptospira spp.) and real-time PCR testing and identified risk factors for exposure. We also used non-targeted metagenomic sequencing on nine selected patients to assess the presence of other possible bacterial causes of non-malarial fevers. Considerable PCR positivity was found for Brucella (19.4%, 95% confidence intervals [CI] 14.2-25.5) and Leptospira spp. (1.7%, 95% CI 0.4-4.9), and high endpoint titres were observed against leptospiral serovar Grippotyphosa from the serological testing. Patients aged 5-17 years old had 4.02 (95% CI 1.18-13.70, p-value = 0.03) and 2.42 (95% CI 1.09-5.34, p-value = 0.03) times higher odds of infection with Brucella spp. and Coxiella burnetii than those of ages 35-80. Additionally, patients who sourced water from dams/springs, and other sources (protected wells, boreholes, bottled water, and water pans) had 2.39 (95% CI 1.22-4.68, p-value = 0.01) and 2.24 (1.15-4.35, p-value = 0.02) times higher odds of exposure to C. burnetii than those who used unprotected wells. Streptococcus and Moraxella spp. were determined using metagenomic sequencing. Brucellosis, leptospirosis, Streptococcus and Moraxella infections are potentially important causes of non-malarial fevers in Garissa. This knowledge can guide routine diagnosis, thus helping lower the disease burden and ensure better health outcomes, especially in younger populations.

Accuracy of rapid lateral flow immunoassays for human leptospirosis diagnosis: A systematic review and meta-analysis.

BACKGROUND: In the last two decades, several rapid lateral flow immunoassays (LFIs) for the diagnosis of human leptospirosis were developed and commercialized. However, the accuracy and reliability of these LFIs are not well understood. In this study, we aimed to evaluate the accuracy of leptospirosis LFIs as well as the factors affecting the test efficiency using systematic review and meta-analysis. METHODS AND RESULTS: Original articles reporting the accuracy of human leptospirosis LFIs against microagglutination tests (MAT) or immunofluorescent assays (IFA) were searched from PubMed, Embase, and Scopus, and selected as per pre-set inclusion and exclusion criteria. A total of 49 data entries extracted from 24 eligible records published between 2003 and 2023 were included for meta-analysis. A meta-analysis was performed using STATA. The quality of the included studies was assessed according to the revised QUADAS-2. Only nine studies (32.1%) were considered to have a low risk of bias and no concern for applicability. Pooled sensitivity and specificity were calculated to be 68% (95% confidence interval, CI: 57-78) and 93% (95% CI: 90-95), respectively. However, the ranges of sensitivity (3.6 - 100%) and specificity (53.5 - 100%) of individual entries are dramatically broad, possibly due to the heterogeneity found in both study designs and LFIs themselves. Subgroup analysis demonstrated that IgM detection has better sensitivity than detection of IgG alone. Moreover, the test performance seems to be unaffected by samples from different phases of infection. CONCLUSIONS: The pooled specificity of LFIs observed is somewhat acceptable, but the pooled sensitivity is low. These results, however, must be interpreted with caution because of substantial heterogeneity. Further evaluations of the LFIs with well-standardized design and reference test will be needed for a greater understanding of the test performance. Additionally, IgM detection type should be employed when leptospirosis LFIs are developed in the future.

Factors associated with differential seropositivity to Leptospira interrogans and Leptospira kirschneri in a high transmission urban setting for leptospirosis in Brazil.

BACKGROUND: Leptospirosis is a zoonosis caused by pathogenic species of bacteria belonging to the genus Leptospira. Most studies infer the epidemiological patterns of a single serogroup or aggregate all serogroups to estimate overall seropositivity, thus not exploring the risks of exposure to distinct serogroups. The present study aims to delineate the demographic, socioeconomic and environmental factors associated with seropositivity of Leptospira serogroup Icterohaemorraghiae and serogroup Cynopteri in an urban high transmission setting for leptospirosis in Brazil. METHODS/PRINCIPAL FINDINGS: We performed a cross-sectional serological study in five informal urban communities in the city of Salvador, Brazil. During the years 2018, 2020 2021, we recruited 2.808 residents and collected blood samples for serological analysis using microagglutination assays. We used a fixed-effect multinomial logistic regression model to identify risk factors associated with seropositivity for each serogroup. Seropositivity to Cynopteri increased with each year of age (OR 1.03; 95% CI 1.01-1.06) and was higher in those living in houses with unplastered walls (exposed brick) (OR 1.68; 95% CI 1.09-2.59) and where cats were present near the household (OR 2.00; 95% CI 1.03-3.88). Seropositivity to Icterohaemorrhagiae also increased with each year of age (OR 1.02; 95% CI 1.01-1.03) and was higher in males (OR 1.51; 95% CI 1.09-2.10), in those with work-related exposures (OR 1.71; 95% CI 1.10-2.66) or who had contact with sewage (OR 1.42; 95% CI 1.00-2.03). Spatial analysis showed differences in distribution of seropositivity to serogroups Icterohaemorrhagiae and Cynopteri within the five districts where study communities were situated. CONCLUSIONS/SIGNIFICANCE: Our data suggest distinct epidemiological patterns associated with the Icterohaemorrhagiae and Cynopteri serogroups in the urban environment at high risk for leptospirosis and with differences in spatial niches. We emphasize the need for studies that accurately identify the different pathogenic serogroups that circulate and infect residents of low-income areas.

Current knowledge on leptospirosis in cats: A systematic review with metanalysis on direct detection, serological response, and clinical data.

Leptospirosis is a neglected bacterial zoonosis that affects a wide range of mammals, with important implications from a One Health perspective. Over the past years feline leptospirosis has gained increased attention in the scientific community. Here we describe a systematic review with meta-analysis that followed the PRISMA guidelines, with an additional PROSPERO registration. The study provides global seropositivity, urinary shedding rates, global serogroup distribution, descriptive data of leptospires that had been isolated from cats and clinical and laboratory features presented by symptomatic cats with acute disease. The search was carried out in six different databases, with the identification of 79 reports describing leptospiral infection in cats. The pooled frequency of seroreactive cats was 11% (95% CI: 9%-13%), with Javanica and Pomona as the most frequent serogroups found. Frequency for urinary shedding was 8% (95% CI: 5%-10%), with L. interrogans identified in most samples. A total of 16 isolates were isolated from cats, with Bataviae as the most frequent serogroup. Twenty symptomatic cats with confirmed leptospiral infection were identified. Anorexia, lethargy, polydipsia, and bleeding disorders were the clinical signs most frequently reported. The results suggest that cats from some locations are exposed to leptospires and may act as urinary shedders of this pathogen, thus indicating a possible role of this species in disease transmission. Clinical data indicates that acute infection is mostly atypical when compared to dogs, and due to difficulties to define an archetypal clinical presentation in cats, feline leptospirosis is likely to be underdiagnosed disease in this species.

Utility of demographic and clinical signs as diagnostic predictors for leptospiral uveitis: A retrospective study.

PURPOSE: Leptospirosis is a waterborne zoonotic disease prevalent in tropical regions, causing significant morbidity and mortality. It can involve any organ in its primary stage, and uveitis is its late complication. While advanced laboratory diagnosis is available only in tertiary care centers globally, a cost-effective bedside assessment of clinical signs and their scoring could offer a provisional diagnosis. AIM: To analyze the diagnostic potential of demographic and clinical signs in a large cohort of serologically confirmed leptospiral uveitis patients. METHODS: In this retrospective study, demographic and clinical parameters of 876 seropositive leptospiral uveitis patients and 1042 nonleptospiral uveitis controls were studied. Multivariable logistic regression analysis with bootstrap confidence interval (CI) characterized the diagnostic predictors. The performance of the model was evaluated using the area under the receiver operating curve (AUROC). RESULTS: Presence of nongranulomatous uveitis (odds ratio [OR] = 6.9), hypopyon (OR = 4.6), vitreous infiltration with membranous opacities (OR = 4.3), bilateral involvement (OR = 4), panuveitis (OR = 3.3), vasculitis (OR = 1.9), disc hyperemia (OR = 1.6), absence of retinochoroiditis (OR = 15), and absence of cystoid macular edema (OR = 8.9) emerged as predictive parameters. The AUROC value was 0.86 with 95% CI of 0.846-0.874. At a cut-off score of 40, the sensitivity and specificity were 79.5 and 78.4, respectively. CONCLUSION: The study demonstrates that ocular signs can serve as diagnostic predictors for leptospiral uveitis, enabling primary care ophthalmologists to make bedside diagnosis. This can be further confirmed by laboratory methods available at tertiary care centers.

Genital route in experimental infection, a promising approach to study genital leptospirosis in ruminants.

Infection by Leptospira sp., mainly strains from the Sejroe serogroup, impairs the reproductive efficiency of ruminants leading to economic losses. Although the majority of experimental studies use the intraperitoneal route of leptospiral infection, it has been suggested that natural infection occurs frequently by sexual transmission. Thus, we assessed the genital route of infection to study genital leptospirosis in the sheep model. A strain of L. borgpetersenii serogroup Sejroe, serovar Hardjobovis was inoculated in 18 ewes, divided into three groups for inoculation: intraperitoneal (n=6; Gip), cervical superficial (genital) (n=6; Ggen) and conjunctival (n=6; Gconj). Monthly, for 90 days, blood samples were collected for serology (MAT) and PCR was performed on urine, cervical-vaginal mucus, and uterine fragments. All ewes were successfully infected, independently of the infection route. Gip and Ggen did not differ throughout the experiment, either on seroconversion or on PCR positivity on urine or genital samples. In contrast, Gconj presented fewer seroreactive animals (P<0.05) and fewer PCR-pos on genital samples than the other groups. The results obtained demonstrated that, although all groups presented both urinary and genital infections, the genital route was more efficient and did not differ from the traditional intraperitoneal. It indicates that genital via, besides being a naturally occurring transmission via, represents a promising and interesting route regarding future studies related to genital leptospirosis in ruminants, and its use should be encouraged.