RESUMO

Leptospira spp. infection is a worldwide zoonosis that causes economic losses to goat rearing, mainly due to reproductive disorders. Hence, the objective of this research was to determine the seroprevalence and associated risk factors of leptospirosis in a goat milk-producing region in the states of Paraiba and Pernambuco, Northeast Brazil. The microscopic agglutination test was used as serological method and risk factor analysis was carried out using univariable and multivariable analyses. Out of the 937 animals sampled, 102 (10.9 %; 95 % CI = 8.9-12.9 %) were seropositive for Leptospira spp. and the most frequent serogroups were Ballum (41.2 %; 95 % CI = 31.6-50.7 %), Icterohaemorrhagiae (25.5 %, 95 % CI = 17-33.9 %) and Semaranga (23.5 %, 95 % CI = 15.3-31.8 %), with antibody titres ranging from 1:50 to 1:200. Thirty-four of 51 herds (66.7 %; 95 % CI = 53.7-79.6 %) had at least one seropositive animal. The risk factor identified in the multivariable two-level random effect binary logistic regression was the animal being an adult (odds ratio = 4.2; 95 % CI = 1.93-9.13; P < 0.001). Our results provide important information on the epidemiology and risk factors associated with goat leptospirosis seroprevalence in one of the main Brazilian goat milk-producing regions. Furthermore, the need for adopting sanitary control measures, especially those involving sanitary management practices, is highlighted.

RESUMO

Leptospirosis and toxoplasmosis are re-emerging zoonosis caused by infection with pathogenic spirochaetes of Leptospira and the protozoa Toxoplasma gondii, respectively. Wild boars (Sus scrofa), an exotic invasive species in Brazil, could play a role in the diseases' epidemiological cycles, but this issue is still unexplored. This study aimed to evaluate the Leptospira spp. and T. gondii seropositivity in wild boars in Rio Grande do Sul state, south Brazil. Of evaluated animals, 16% (13/80) and 85% (52/61) had antibodies to T. gondii and Leptospira spp., respectively. Sex, weight, age, hunt location and season of hunt were evaluated by their association with seropositivity for both pathogens, but none of them had statistical significance. This study revealed that wild boars should be considered as a potential source of Leptospira spp. and T. gondii dissemination for humans and animal species in shared environments in Rio Grande do Sul state.

Assuntos

Anticorpos Antiprotozoários , Leptospira , Leptospirose , Sus scrofa , Doenças dos Suínos , Toxoplasma , Toxoplasmose Animal , Animais , Leptospirose/epidemiologia , Leptospirose/veterinária , Brasil/epidemiologia , Toxoplasmose Animal/epidemiologia , Toxoplasmose Animal/imunologia , Toxoplasma/imunologia , Leptospira/imunologia , Sus scrofa/parasitologia , Suínos , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/parasitologia , Doenças dos Suínos/microbiologia , Anticorpos Antiprotozoários/sangue , Masculino , Feminino , Estudos Soroepidemiológicos , Anticorpos Antibacterianos/sangue , Zoonoses/epidemiologia , Zoonoses/parasitologia , Humanos , Saúde Pública

RESUMO

Bovine Genital Leptospirosis (BGL) is a chronic reproductive syndrome characterized by genital infection by Leptospira spp. An accurate diagnosis of BGL is crucial to implementing proper control measures in field conditions. This study aimed to evaluate the reliability of serology by Microscopic Agglutination Test (MAT) for diagnosing leptospirosis in subfertile cows with genital infection. Of three herds, 93 non-pregnant cows with reproductive failures were submitted to the blood sampling (serology by MAT) and genital samples (lipL32-PCR). A total of 62/93 (66.6%) cows presented seroreactive to cutoff 100, while 45/93 (48.4%) cows were positive to cutoff 200, mainly against the Sejroe serogroup. In PCR analysis, 55/93 (59.1%) were positive. MAT results were compared with PCR (considered the standard), and test parameters and Cohen's kappa (ƙ) were calculated for the cut-offs 100 and 200. A ROC curve was performed for each cut-off of titers 100 to 1,600. The sensitivity and specificity of MAT100 were calculated at 66.6% and 33.3%, while for MAT200 the sensitivity was estimated as 35% and specificity as 54.5%. The accuracy of MAT was poor, being 54.8% in MAT100 and 42% in MAT200. Furthermore, the area under the curve of ROC analysis was low for all titers, and the correlation was poor for MAT100 and MAT200 (ƙ < 0). The results demonstrated that MAT is a limited technique to diagnose bovine genital carriers individually, and if only MAT is applied, genital carriers may pass undetected, impairing the control programs.

RESUMO

Bacterial ubiquitous Toxin-Antitoxin (TA) systems are considered to be important survival mechanisms during stress conditions. In regular environmental conditions, the antitoxin blocks the toxin, whereas during imbalanced conditions, the antitoxin concentration decreases, exposing the bacteria cell to a range of toxic events. The most evident consequence of this disequilibrium is cell growth arrest, which is the reason why TAs are generally described as active in the function of bacterial growth kinetics. Virulence-associated proteins B and C (VapBC) are a family of type II TA system, in which VapC is predicted to display the toxic ribonuclease activity while VapB counteracts this activity. Previously, using in silico data, we designated four VapBC TA modules in Leptospira interrogans serovar Copenhageni, the main etiological agent of human leptospirosis in Brazil. The present study aimed to obtain the proteins and functionally characterize the VapBC-1 module. The expression of the toxin gene vapC in E. coli did not decrease the cell growth rate in broth culture, as was expected to happen within active TA modules. However, interestingly, when the expression of the toxin was compared to that of the complexed toxin and antitoxin, cell viability was strongly affected, with a decrease of three orders of magnitude in colony forming unity (CFU). The assumption of the affinity between the toxin and the antitoxin was confirmed in vivo through the observation of their co-purification from cultivation of E. coli co-expressing vapB-vapC genes. RNAse activity assays showed that VapC-1 cleaves MS2 RNA and ribosomal RNA from L. interrogans. Our results indicate that the VapBC-1 module is a potentially functional TA system acting on targets that involve specific functions. It is very important to emphasize that the common attribution of the functionality of TA modules cannot be defined based merely on their ability to inhibit bacterial growth in a liquid medium.

RESUMO

Leptospira is a bacterial genus that includes several pathogenic species related to leptospirosis. In Colombia, leptospirosis is a mandatorily reported disease, widely distributed across the country. In the Villeta municipality, leptospirosis has been identified as an important cause of febrile illness; however, to date, no studies have been performed to identify the circulating species. A genus-specific qualitative qPCR was performed on DNA extracted from febrile patients' acute-phase whole-blood samples targeting a fragment of the rrs gene. Positive qPCR samples were further amplified for the adk, icdA, LipL32, LipL41, rrs, and secY genes through conventional PCR for sequencing. All high-quality obtained sequences were further assessed through concatenated phylogenetic analysis. A total of 25% (14/56) of febrile patients' acute blood samples were positive for Leptospira spp. High-quality sequences were obtained for only five genes, and analysis through concatenated phylogeny identified that all sequences clustered within the P1/pathogenic clade; some of them formed a robustly supported clade with Leptospira santarosai, and others were closely related with other Leptospira species but exhibited considerable genetic divergence. We describe the presence of pathogenic Leptospira species among febrile patients from the Villeta municipality and identify L. santarosai and other Leptospira species as causative agents of leptospirosis in the region.

RESUMO

Leptospirosis is a zoonotic disease whose transmission is linked to multiple factors involving the interface between animals, humans, and the environment. This disease is of great importance for public health, as it profoundly affects the health aspects of the population and animals. Considering the importance of non-human primates in this epidemiological chain, the objective of this research was to conduct a systematic literature review with meta-analysis, providing information on leptospirosis in non-human primates (NHPs) and an update on the infection situation in Brazil and other countries. Thus, a search was performed in five databases, initially finding 3332 studies, of which 32 met the eligibility criteria and were used for the systematic review. According to them, the most prevalent serogroup in non-human primates was Icterohaemorrhagiae, which is adapted to rodents as primary hosts. A wide distribution of the infection was found in the regions of both wild and captive animals. Through meta-analysis, the seroprevalence rate of leptospirosis in non-human primates was 27.21% (CI 17.97-38.95%). Cochran's Q test (p < 0.01) identified heterogeneity between studies, classified as high by the Higgins and Thompson test (I2 = 92.4%). Therefore, seroepidemiological and Leptospira isolation studies in non-human primates are important to investigate and monitor the suspected impact of these species as maintainers or transmitters of the pathogen to humans and other wild and domestic animals, in addition to demonstrating the need for standardization related to control and prevention measures.

Assuntos

Leptospirose , Primatas , Animais , Leptospira/imunologia , Leptospira/isolamento & purificação , Leptospirose/sangue , Leptospirose/epidemiologia , Leptospirose/microbiologia , Leptospirose/veterinária , Doenças dos Macacos/sangue , Doenças dos Macacos/epidemiologia , Doenças dos Macacos/microbiologia , Doenças dos Macacos/transmissão , Primatas/microbiologia , Estudos Soroepidemiológicos , Zoonoses/sangue , Zoonoses/epidemiologia , Zoonoses/microbiologia , Zoonoses/transmissão , Humanos

RESUMO

Leptospirosis, classified by the World Health Organization as an emerging and neglected disease, is caused by the zoonotic pathogen Leptospira interrogans. This review aims to outline the Mexican epidemic of L. interrogans in dogs, including diagnosis and prevention methods. This review article searched articles from the publishers Wiley, Springer, PubMed, Redalyc, SciElo, and Elsevier. Among the 200 Mexican articles concerning Leptospira epidemiology, diagnosis, treatment, and vaccination, those that failed to meet the set inclusion criteria were excluded. The worldwide study of L. interrogans has focused on this bacterium. In Mexico, up-to-date information on canine prevalence, diagnosis, and vaccine use is scarce. Flow cytometrically detected Salmonella serovars differ from those in current vaccines, emphasizing the importance of broadening vaccine serovar coverage.

RESUMO

Leptospirosis is a global zoonotic disease caused by spirochete bacteria of the genus Leptospira. The disease exhibits a notable incidence in tropical and developing countries, and in Colombia, environmental, economic, social, and cultural conditions favor disease transmission, directly impacting both mortality and morbidity rates. Our objective was to establish the pooled lagged effect of runoff on leptospirosis cases in Colombia. For our study, we included the top 20 Colombian municipalities with the highest number of leptospirosis cases. Monthly cases of leptospirosis, confirmed by laboratory tests and spanning from 2007 to 2022, were obtained from the National Public Health Surveillance System. Additionally, we collected monthly runoff and atmospheric and oceanic data from remote sensors. Multidimensional poverty index values for each municipality were sourced from the Terridata repository. We employed causal inference and distributed lag nonlinear models to estimate the lagged effect of runoff on leptospirosis cases. Municipality-specific estimates were combined through meta-analysis to derive a single estimate for all municipalities under study. The pooled results for the 20 municipalities suggest a lagged effect for the 0 to 2, and 0-3 months of runoff on leptospirosis when the runoff is < 120 g/m2. No effect was identified for longer lagged periods (0-1, 0 to 4, 0 to 5, and 0-6 months) or higher runoff values. Incorporation of the multidimensional poverty index into the meta-analysis of runoff contributed to the models for the lagged periods of 0-3, and 0-4 months.

RESUMO

Leptospirosis is a common but underdiagnosed zoonosis. We conducted a 1-year prospective study in La Guaira State, Venezuela, analyzing 71 hospitalized patients who had possible leptospirosis and sampling local rodents and dairy cows. Leptospira rrs gene PCR test results were positive in blood or urine samples from 37/71 patients. Leptospira spp. were isolated from cultured blood or urine samples of 36/71 patients; 29 had L. interrogans, 3 L. noguchii, and 4 L. venezuelensis. Conjunctival suffusion was the most distinguishing clinical sign, many patients had liver involvement, and 8/30 patients with L. interrogans infections died. The Leptospira spp. found in humans were also isolated from local rodents; L. interrogans and L. venezuelensis were isolated from cows on a nearby, rodent-infested farm. Phylogenetic clustering of L. venezuelensis isolates suggested a recently expanded outbreak strain spread by rodents. Increased awareness of leptospirosis prevalence and rapid diagnostic tests are needed to improve patient outcomes.

Assuntos

Surtos de Doenças , Leptospira , Leptospirose , Filogenia , Roedores , Animais , Leptospirose/epidemiologia , Leptospirose/veterinária , Leptospirose/microbiologia , Leptospirose/diagnóstico , Humanos , Venezuela/epidemiologia , Bovinos , Leptospira/genética , Leptospira/isolamento & purificação , Leptospira/classificação , Feminino , Roedores/microbiologia , Adulto , Masculino , Pessoa de Meia-Idade , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/epidemiologia , Adolescente , Leptospira interrogans/genética , Leptospira interrogans/isolamento & purificação , Leptospira interrogans/classificação , Adulto Jovem , Estudos Prospectivos , Criança , Idoso , Doenças Endêmicas , Zoonoses/epidemiologia , Zoonoses/microbiologia , Pré-Escolar

RESUMO

AIMS: Currently, immunoinformatic approaches have shown promise in rapidly and cost-effectively identifying new antigens from the Leptospira proteome. Chimeric multiepitope proteins offer a strategy with significant potential for implementation in diagnosis and vaccines development. METHODS AND RESULTS: In this study, we detail the immunoinformatic analyses and design of a new recombinant chimeric protein constructed with epitopes identified from the sequences of ErpY-like and LemA proteins, previously identified as potential antigens for controlling leptospirosis. We expressed the chimeric protein using Escherichia coli heterologous systems, evaluated its antigenicity using serum from naturally infected patients, and its immunogenicity in mice as an animal model, with Freund as an adjuvant. The resulting recombinant chimeric protein, named rErpY-LemA, was successfully expressed and purified using a prokaryotic system, with an expected mass of 35 kDa. Serologic assays using serum samples from naturally infected patients demonstrated recognition of the chimera protein by antibodies present in sera. Animals immunized with the chimera exhibited a significant IgG antibody response from the 7th day (P < 0.001), persisting until day 49 of experimentation, with a titer of 1:12,800 (P < 0.05). Notably, significant production of IgA, IgM, and IgG subclasses was observed in animals immunized with the chimera. CONCLUSIONS: These results highlight the promising role of immunoinformatics in rapidly identifying antigens and the potential of chimeric multiepitope proteins in developing effective strategies for leptospirosis control.

Assuntos

Antígenos de Bactérias , Leptospirose , Proteínas Recombinantes de Fusão , Leptospirose/imunologia , Leptospirose/prevenção & controle , Animais , Camundongos , Antígenos de Bactérias/imunologia , Antígenos de Bactérias/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/genética , Humanos , Anticorpos Antibacterianos/sangue , Leptospira/imunologia , Leptospira/genética , Biologia Computacional , Epitopos/imunologia , Epitopos/genética , Vacinas Bacterianas/imunologia , Vacinas Bacterianas/genética , Escherichia coli/genética , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/genética , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/genética

RESUMO

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.

Assuntos

Anticorpos Antibacterianos , Vacinas Bacterianas , Leptospirose , Lipoproteínas , Animais , Leptospirose/prevenção & controle , Leptospirose/imunologia , Lipoproteínas/imunologia , Lipoproteínas/genética , Vacinas Bacterianas/imunologia , Vacinas Bacterianas/genética , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Cricetinae , Epitopos de Linfócito B/imunologia , Epitopos de Linfócito B/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/genética , Adjuvantes Imunológicos/administração & dosagem , Imunoglobulina G/sangue , Epitopos de Linfócito T/imunologia , Epitopos de Linfócito T/genética , Leptospira interrogans/imunologia , Leptospira interrogans/genética , Proteínas da Membrana Bacteriana Externa/imunologia , Proteínas da Membrana Bacteriana Externa/genética , Vacinação , Imunidade Humoral , Leptospira/imunologia , Leptospira/genética , Imunogenicidade da Vacina

RESUMO

Background: Leptospira is a genus of bacteria that causes the zoonotic disease known as leptospirosis, which mainly affects countries with tropical and subtropical climates. Its prevalence may be underestimated because the initial stage of the infection is characterized by presenting a febrile condition that is easily confused with other diseases, such as dengue. This work reports the frequency of leptospirosis in the blood of patients with febrile symptoms of unknown origin. Materials and Methods: A total of 218 peripheral blood samples were analyzed from volunteer participants from Culiacan Sinaloa in June 2019, one half corresponded to patients with undiagnosed febrile symptoms and the other half to asymptomatic volunteers. Data collected included the age and sex of the participants. Leptospira was detected by qPCR using a fragment of the lipL32 gene from the bacteria's genome as a target. Fisher's exact test was used as a statistical method to estimate the relationship between the infection and the data collected. Results: The study group comprised 134 female and 84 male patients ranging from ages 1 to 92 years, averaging 41 years. In this study, Leptospira infection was identified in the blood of 22/218 participating volunteers (10.09%), of which 20/109 (18.34%) presented febrile symptoms, whereas 2/109 (1.83%) were asymptomatic. The most affected participants were women with ages between 27 and 59 years. However, the analysis of the relationship between infection and the variables studied did not show statistical significance. Conclusions: Leptospirosis was detected in blood samples from patients with undiagnosed febrile illness and asymptomatic symptoms in Sinaloa. The lipL32 gene is useful as a target in identifying Leptospira in human blood in the acute phase of the disease.

RESUMO

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.

Assuntos

Leptospira , Leptospirose , Infecções por Rickettsia , Rickettsia , Humanos , Peru/epidemiologia , Rickettsia/isolamento & purificação , Feminino , Masculino , Leptospira/isolamento & purificação , Leptospira/patogenicidade , Leptospirose/epidemiologia , Leptospirose/microbiologia , Leptospirose/complicações , Leptospirose/diagnóstico , Infecções por Rickettsia/epidemiologia , Infecções por Rickettsia/microbiologia , Infecções por Rickettsia/diagnóstico , Adulto , Animais , Febre/microbiologia , Zoonoses/microbiologia , Zoonoses/diagnóstico , Zoonoses/epidemiologia , Mialgia/microbiologia , Mialgia/epidemiologia , Pessoa de Meia-Idade , Adulto Jovem , Adolescente , Cefaleia/microbiologia , Cefaleia/etiologia , Cefaleia/epidemiologia , Artralgia/microbiologia , Artralgia/etiologia

RESUMO

Background and Aim: Leptospirosis is a re-emerging zoonosis that is under-reported in tropical countries, and canines can be a potential reservoir of the disease. The objective of this study was to diagnose Leptospira spp. that is actively infected and re-infected in stray dogs and cats from Bogota, D.C., Colombia. Materials and Methods: A sample of 200 animals, including dogs and cats from the animal protection programs of Bogota, Colombia, were used in this study. Blood was collected from these animals for serum and DNA analysis. Conventional polymerase chain reaction (PCR) was performed using the 16s rRNA primer set, and higher-quality amplification products were sequenced by Sanger. For serodiagnosis, a group of PCR-positive samples was tested using the microagglutination test (MAT). Results: The overall PCR positivity of stray dogs and cats was 56%, 52.9%, and 65.3% in dogs and cats, respectively. The MAT seropositivity was 77.3%, and only dogs showed titers higher than 1:400. Canicola, Icterohaemorrhagiae, Pomona, Hardjo Prajitno, and Canicola and Hardjo prajitno were the serogroups associated with dogs and cats, respectively. Phylogenetic analysis revealed that the strains belonging to Leptospira interrogans serovars related to isolated samples of American, European, and Asian bats (Myotis myotis), dogs, and bovines of American origin. Conclusion: These results showed that stray dogs and cats were previously exposed to different serovars of Leptospira spp. and re-infected with other serovars that actively participated in the transmission cycle. These findings highlight the importance of actively diagnosing infectious animals to design effective intervention strategies.

RESUMO

Leptospirosis is an infectious disease that affects domestic animals, wild animals, and humans. It represents a public health problem and has an important economic impact on livestock. This study aims to investigate the importance of genital and transplacental infection in the epidemiology of leptospirosis in cows maintained in Caatinga biome conditions, Northeastern Brazil, as well as reporting organs colonized by Leptospira spp. in embryos and fetuses. Blood, urinary tract (urine, bladder, and kidney), and reproductive tract (vaginal fluid, uterus, uterine tube, ovary, and placenta) samples were collected from 15 slaughtered pregnant cows. Two embryos and 13 fetuses were sampled. Central nervous system and choroid ovoid samples were collected from embryos. Blood, central nervous system, lung, peritoneal liquid, abomasal content, liver, spleen, urine, bladder, kidney, and reproductive system samples were collected from fetuses. Diagnostic methods included the microscopic agglutination test (MAT) using a collection of 24 serovars belonging to 17 different pathogenic serogroups of five species as antigens, as well as polymerase chain reaction (PCR). Anti-Leptospira spp. antibodies were found in 9 cows (60%), while 13 cows (86.67%) had at least one organ or urine with leptospiral DNA. No fetus was seroreactive. Among the embryos and fetuses, 13 (86.67%) presented leptospiral DNA, proving a high frequency of transplacental infection (100%). For cows, the most frequent biological materials regarding Leptospira spp. DNA detection were placenta (13 out of 15 samples; 86.7%), uterus (10 out of 15 samples; 66.7%), and vaginal fluid (5 out of 15 samples; 33.3%), while, for fetuses/embryos, the most frequent PCR-positive samples were choroid ovoid (1/2; 50%), spleen (6/13; 46.2%), kidney (5/13; 38.5%), and central nervous system (5/15; 33.3%). Sequenced samples based on the LipL32 gene presented 99% similarity with L. borgpetersenii. The results indicate that transplacental infection is an efficient way of spreading Leptospira spp. in cows maintained in Caatinga biome conditions. Therefore, prevention and control strategies must include actions that interrupt transmission through this alternative route.

RESUMO

Leptospira spp. are bacteria responsible for leptospirosis, a zoonotic disease with considerable impacts on the economy, animal health, and public health. This disease has a global distribution and is particularly prevalent in Brazil. Both rural and urban environments are habitats for Leptospira spp., which are primarily transmitted through contact with the urine of infected animals. Consequently, domestic and wild species can harbor these prokaryotes and serve as infection sources for other hosts. In the context of wild animals, there is a dearth of molecular studies elucidating the roles of various animal and bacterial species in the epidemiology of leptospirosis. Therefore, this study aimed to evaluate the presence of Leptospira spp. DNA in different species of free-living and captive wild animals and to assess the phylogenetic relationships of the identified microorganisms in Rio Grande do Sul, Brazil. The samples were evaluated for the presence of the gene lipL32 by polymerase chain reaction (PCR) and sequencing of the amplified fragment after which phylogenetic analyzes were carried out. DNA from Leptospira spp. was extracted from kidney tissue from wild animals (Mammalia class). Pathogenic Leptospira spp. DNA was detected in 9.6% (11/114) of the samples, originating from nine species of wild animals, including the white-eared opossum (Didelphis albiventris), skunk (Conepatus chinga), geoffroy's cat (Leopardus geoffroyi), margay (Leopardus wiedii), pampas fox (Lycalopex gymnocercus), capybara (Hydrochoerus hydrochaeris), common marmoset (Callithrix jacchus), neotropical river otter (Lontra longicaudis), and european hare (Lepus europaeus). Phylogenetic analysis revealed the presence of Leptospira borgpetersenii and Leptospira interrogans in these animals. This research is the first study contributing to the epidemiology of leptospirosis by identifying L. borgpetersenii and L. interrogans in free-living and captive wild animals in Rio Grande do Sul, Brazil, potentially acting as bacterial reservoirs. Additionally, our findings can inform sanitary measures for controlling and preventing the disease, thereby safeguarding public health.

Assuntos

Animais Selvagens , Leptospira interrogans , Leptospira , Leptospirose , Filogenia , Animais , Brasil/epidemiologia , Leptospirose/microbiologia , Leptospirose/veterinária , Leptospirose/epidemiologia , Animais Selvagens/microbiologia , Leptospira/genética , Leptospira/isolamento & purificação , Leptospira/classificação , Leptospira interrogans/genética , Leptospira interrogans/classificação , Leptospira interrogans/isolamento & purificação , Mamíferos/microbiologia , DNA Bacteriano/genética

RESUMO

Leptospirosis is a neglected zoonotic disease that commonly affects cattle, pigs, horses, and dogs in many countries. Infection in dogs is usually subclinical, but acute cases of leptospirosis may occur along with systemic failure, which may become fatal. After recovery from an acute infection, dogs may become asymptomatic carriers and shed pathogenic leptospires through urine for long periods of time. Here, a study of ten different cases of leptospirosis is presented, showing the relevance of dogs as asymptomatic carriers of pathogenic Leptospira. The diagnosis was confirmed via isolation and further serological and genetic identification. Four Leptospira isolates (LOCaS28, 31, 34, and 46) were obtained from the kidneys and urine samples of 58 dogs destined for destruction (6.89%) at a Canine Control Center in Mexico City. No spirochetes were observed in the urine samples of those Leptospira-positive dogs examined under dark-field microscopy, and no clinical signs of disease were observed either. Six additional isolates were obtained: two came from asymptomatic carrier dogs (CEL60 and UADY22); another isolate came from an asymptomatic dog that was a pack companion of a clinically ill dog with fatal leptospirosis (AGFA24); and finally, three isolates were taken from dogs that died of leptospirosis (LOCaS59, Citlalli, and Nayar1). Nine out of the ten isolates were identified as being from the serogroup Canicola via cross-absorption MAT using reference strains and specific antisera, and their identity was genetically confirmed as Canicola ST34 via multi-locus sequencing typing (MLST). In contrast, the isolate Nayar1 was identified as serovar Copenhageni ST2. Interestingly, the asymptomatic dogs from which Leptospira isolates were recovered consistently showed high antibody titers in the microscopic agglutination test (MAT), revealing values of at least 1:3200 against serogroup Canicola and lower titer values against other serogroups. Isolates showed different virulence levels in the hamster model. Taken as a whole, all these findings confirmed that dogs may act as asymptomatic carriers of pathogenic leptospires and possibly spread them out to the environment, thus representing an active public health risk. The results also showed that the Canicola ST34 clone is the most prevalent Leptospira serovar in dogs in Mexico, and finally that the old-fashioned MAT is a good alternative for the detection of presumptive Leptospira asymptomatic carrier dogs.

RESUMO

Introducción: La leptospirosis es una zoonosis que cons-tituye un problema emergente de salud pública. La insufi-ciencia renal, plaquetopenia y compromiso respiratorio se describen como predictores de mortalidad.Objetivos: Describir características clínicas, radiológicas y de laboratorio de individuos hospitalizados por leptos-pirosis y evaluar los predictores de mala evolución clínica (MEC).Materiales y métodos: Estudio de cohorte de inclusión ambispectiva de pacientes con leptospirosis internados en un hospital de la ciudad de Santa Fe entre 1997 y 2022. Se definió MEC como la admisión a Unidad de Cuidados Intensivos (UCI), requerimiento de asistencia respiratoria mecánica (ARM) y/o muerte. Se utilizaron las pruebas de Chi2, test T de Student o la U de Mann-Whitney, según co-rrespondiera. Se construyó una regresión logística binaria con las variables con p<0,05.Resultados: 101 pacientes, 87,1% (n=88) hombres, media-na de edad de 29 (RIC 20-44) años. La fiebre fue el síntoma más frecuente [83,2% (n=84)], seguido del compromiso di-gestivo [62,4% (n=63)]. Las alteraciones de laboratorio más frecuentes fueron: eritrosedimentación elevada [91,9% (n=79)] y leucocitosis [61% (n=61)]. Se observó MEC en el 25,7% (n=26). El 25,7% (n=26) fue admitido en UCI, el 13,9% (n=14) requirió ARM y el 5% (n=5) falleció. La presencia de plaquetopenia (OR=13,3, IC95% 2-80), las alteraciones en la radiografía de tórax (OR=33,5, IC95% 5-225) y la ausencia de cefalea (OR=6,8, IC95% 1-32) fueron predictores inde-pendientes de MEC.Conclusiones: En concordancia con la bibliografía, la afec-tación pulmonar y plaquetopenia son factores de riesgo para la mala evolución clínica. En nuestra serie, la cefalea constituyó un síntoma protector

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Introduction: Leptospirosis is an emerging zoonotic di-sease that poses a public health problem. Renal failu-re, thrombocytopenia, and respiratory involvement have been described as predictors of mortality.Objectives: To describe the clinical, radiological, and la-boratory characteristics of hospitalized individuals with leptospirosis and evaluate predictors of poor clinical outcomes (PCO).Materials and methods: A prospective cohort study was conducted including patients with leptospirosis admit-ted to a hospital in the city of Santa Fe between 1997 and 2022. PCO was defined as admission to the Intensive Care Unit (ICU), requirement for mechanical respiratory assistance (MRA), and/or death. The chi-square test, Student>s t-test, or Mann-Whitney U test were used as appropriate. A binary logistic regression was performed with variables having p<0.05.Results: Out of the 101 patients included, 87.1% (n=88) were male, with a median age of 29 (IQR 20-44) years. Fever was the most common symptom [83.2% (n=84)], followed by digestive involvement [62.4% (n=63)]. The most frequent laboratory abnormalities were elevated erythrocyte sedimentation rate [91.9% (n=79)] and leuko-cytosis [61% (n=61)]. PCO was observed in 25.7% (n=26) of patients, with 25.7% (n=26) admitted to the ICU, 13.9% (n=14) requiring MRA, and 5% (n=5) resulting in death. The presence of thrombocytopenia (OR=13.3, 95% CI 2-80), abnormalities in chest X-rays (OR=33.5, 95% CI 5-225), and absence of headache (OR=6.8, 95% CI 1-32) were predictors of PCO. Conclusions: Consistent with the literature, pulmonary involvement and thrombocytopenia are independent risk factors for poor clinical outcomes. In our series, the pre-sence of headache was a protective symptom

Assuntos

Humanos , Masculino , Feminino , Doenças Endêmicas/prevenção & controle , Hospitalização , Leptospira/patogenicidade , Leptospirose/mortalidade

RESUMO

INTRODUCTION: Bats are a diverse group of mammals that have unique features allowing them to act as reservoir hosts for several zoonotic pathogens such as Leptospira. Leptospires have been classified into pathogenic, intermediate, and saprophytic groups and more recently into clades P1, P2, S1, and S2, being all the most important pathogenic species related to leptospirosis included within the P1/pathogenic clade. Leptospira has been detected from bats in several regions worldwide; however, the diversity of leptospires harboured by bats is still unknown. AIM: The aim of the present study was to determine the genetic diversity of Leptospira spp. harboured by bats worldwide. METHODS: A systematic review was conducted on four databases to retrieve studies in which Leptospira was detected from bats. All studies were screened to retrieve all available Leptospira spp. 16S rRNA sequences from the GenBank database and data regarding their origin. Sequences obtained were compared with each other and reference sequences of Leptospira species and analysed through phylogenetic analysis. RESULTS: A total of 418 Leptospira spp. 16S rRNA sequences isolated from 55 bat species from 14 countries were retrieved from 15 selected manuscripts. From these, 417 sequences clustered within the P1/pathogenic group, and only one sequence clustered within the P2/intermediate group. Six major clades of P1/pathogenic Leptospira spp. were identified, three of them composed exclusively of sequences obtained from bats. CONCLUSION: We identified that bats harbour a great genetic diversity of Leptospira spp. that form part of the P1/pathogenic clade, some of which are closely related to leptospirosis-associated species. This finding contributes to the knowledge of the diversity of leptospires hosted by bats worldwide and reinforces the role of bats as reservoirs of P1/pathogenic Leptospira spp.

Assuntos

Quirópteros , Variação Genética , Leptospira , Leptospirose , Filogenia , Animais , Quirópteros/microbiologia , Leptospira/genética , Leptospira/classificação , Leptospira/isolamento & purificação , Leptospirose/veterinária , Leptospirose/microbiologia , Leptospirose/epidemiologia , Reservatórios de Doenças/veterinária , Reservatórios de Doenças/microbiologia , RNA Ribossômico 16S/genética , Zoonoses

RESUMO

Leptospira is a genus of bacteria that includes free-living saprophytic species found in water or soil, and pathogenic species, which are the etiologic agents of leptospirosis. Besides all the efforts, there are only a few proteins described as virulence factors in the pathogenic strain L. interrogans. This work aims to perform L. biflexa serovar Patoc1 strain Paris global proteome and to compare with the proteome database of pathogenic L. interrogans serovar Copenhageni strain Fiocruz L1-130. We identified a total of 2327 expressed proteins of L. biflexa by mass spectrometry. Using the Get Homologues software with the global proteome of L. biflexa and L. interrogans, we found orthologous proteins classified into conserved, low conserved, and specific proteins. Comparative bioinformatic analyses were performed to understand the biological functions of the proteins, subcellular localization, the presence of signal peptide, structural domains, and motifs using public softwares. These results lead to the selection of 182 low conserved within the saprophyte, and 176 specific proteins of L. interrogans. It is anticipated that these findings will indicate further studies to uncover virulence factors in the pathogenic strain. This work presents for the first time the global proteome of saprophytic strain L. biflexa serovar Patoc, strain Patoc1. SIGNIFICANCE: The comparative analysis established an array of specific proteins in pathogenic strain that will narrow down the identification of immune protective proteins that will help fight leptospirosis.

Assuntos

Leptospira interrogans , Leptospira , Leptospirose , Humanos , Proteoma/metabolismo , Fatores de Virulência/metabolismo