Leptospira Infection Research Articles

Low Prevalence of Leptospira Carriage in Rodents in Leptospirosis-Endemic Northeastern Thailand.

Leptospirosis is a neglected zoonotic disease affecting mostly the world’s tropical regions. The rural people of northeastern Thailand suffer from a large number of leptospirosis infections, and their abundant rice fields are optimal rodent habitats. To evaluate the contribution of diversity and carriage rate of pathogenic Leptospira in rodent reservoirs to leptospirosis incidence, we surveyed rodents, between 2011 and 2012, in four provinces in northeastern Thailand with the highest incidence rates of human leptospirosis cases. We used lipL32 real-time PCR to detect pathogenic Leptospira in rodent kidneys, partial 16S rRNA gene sequencing to classify the infecting Leptospira species, and whole 16S rDNA sequencing to classify species of isolated Leptospira. Overall prevalence of Leptospira infection was 3.6% (18/495). Among infected rodents, Bandicota indica (14.3%), Rattus exulans (3.6%), and R. rattus (3.2%) had renal carriage. We identified two pathogenic Leptospira species: L. interrogans (n = 15) and L. borgpetersenii (n = 3). In addition, an L. wolffii (LS0914U) isolate was recovered from the urine of B. indica. Leptospira infection was more prevalent in low density rodent populations, such as B. indica. In contrast, there was a lower prevalence of Leptospira infection in high density rodent populations of R. exulans and R. rattus.

Rodent Control a Tool for Prevention of Leptospirosis - A Success Story on Human Leptospirosis with Gujarat State

Leptospirosis is a debilitating zoonosis for humans as well as domestic animals worldwide and widespread in tropical and sub tropical areas with high rainfall. India being a tropical country is potential for this debilitating disease. Among mammals, rodents are most important and widely distributed reservoirs of leptospiral infection. Four species of rodents i.e., Rattus norvegicus, Rattus rattus, Bandicota indica and Bandicota bengalensis, are known vectors for this disease and transmit the infection in rural and urban areas. Hence, rodent control in larger areas is recommended for preventing the spread of this disease. Gujarat is one of the States having established leptospirosis endemic pockets, particularly in Valsad, Navsari, Tapi and Surat districts. Hence, the Departments of Health and Agriculture, Gujarat conducted coordinated rodent vector control campaigns to prevent the disease spread. All activities were guided by National Institute of Plant Health Management, Hyderabad and funded under National Plan on Rodent Pest Management of Department of Agriculture and Cooperation, Ministry of Agriculture, Government of India. The initial pilot scale rodent control campaigns conducted in 114 villages covering 5,127 hectares jointly by Agriculture and Health Departments of the state in 2009 brought a reduction of 61% leptospirosis disease prevalence. Following this success, large scale community based rodent control campaigns were organized during 2012 in these districts in 1822 villages covering 4,70,782 hectares and obtained 82% reduction in the disease prevalence. It was observed that the inter-departmental coordination of Agriculture and Health Departments brought these significant results, which can be replicated and adopted elsewhere to prevent leptospirosis in the country.

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

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

An historical view of the experimental leptospiral infection in ruminants

The first experimental infections with Leptospira in ruminants were conducted in the 1950s, primarily assessed the pathogenesis caused by serovar Pomona in cows. Throughout the decades, experimental infections have also demonstrated the clinical aspects of the infection by other strains, mainly Hardjo. Despite the important outcomes observed in experimental infections in ruminants, there is still a large discrepancy regarding the ideal dose, route, strain, model species or animal age that should be used to reproduce the acute and chronic leptospirosis in ruminants. In this context, the present study aimed to review the historical processes involved on the experimental leptospiral infection in ruminants. The inclusion criteria were papers that clearly described inoculation route, strain, dose, clinical signs and animal age. Overall, 37 experiments were noted. The most frequently reported clinical signs were fever, prostration, hematuria and death, with the majority of them occurring in young animals infected by incidental strains. Regarding reproductive problems, they occurred in the majority of the experiments and were also more related to incidental strains. In this context, abortions, retained placenta and weak fetuses were the most frequent symptoms. Noteworthy that although the mechanisms of the clinical acute disease either systemic or reproductive, is reasonably well understood, the physiopathology involved on reproductive problems due to the silent chronic infection is less discussed and remains to be elucidated. In this context, it is evident the need for studies focused on the genital infection and reproductive aspects of leptospiral infection in ruminants.

A Comparison of Intramuscular and Subcutaneous Administration of LigA Subunit Vaccine Adjuvanted with Neutral Liposomal Formulation Containing Monophosphoryl Lipid A and QS21.

Leptospirosis vaccines with higher potency and reduced adverse effects are needed for human use. The carboxyl terminal domain of leptospiral immunoglobulin like protein A (LigAc) is currently the most promising candidate antigen for leptospirosis subunit vaccine. However, LigAc-based vaccines were unable to confer sterilizing immunity against Leptospira infection in animal models. Several factors including antigen properties, adjuvant, delivery system, and administration route need optimization to maximize vaccine efficacy. Our previous report demonstrated protective effects of the recombinant LigAc (rLigAc) formulated with liposome-based adjuvant, called LMQ (neutral liposome combined with monophosphoryl lipid A and Quillaja saponaria fraction 21) in hamsters. This study aimed to evaluate the impact of two commonly used administration routes, intramuscular (IM) and subcutaneous (SC), on immunogenicity and protective efficacy of rLigAc-LMQ administrated three times at 2-week interval. Two IM vaccinations triggered significantly higher levels of total anti-rLigAc IgG than two SC injections. However, comparable IgG titers and IgG2/IgG1 ratio was observed for both routes after the third immunization. The route of vaccine administration did not influence the survival rate (60%) and renal colonization against lethal Leptospira challenge. Importantly, the kidneys of IM group showed no pathological lesions while the SC group showed mild damage. In conclusion, IM vaccination with rLigAc-LMQ not only elicited faster antibody production but also protected from kidney damage following leptospiral infection better than SC immunization. However, both tested routes did not influence protective efficacy in terms of survival rate and the level of renal colonization.

Potential use of a canine whole blood culture system to evaluate the immune response to Leptospira

In susceptible hosts, protection from Leptospira infection is mediated by the innate immune response at the point of entry and humoral immunity. Thus, identifying and segregating the initial host response at the representative host-pathogen interface is needed to understand the typical outcomes of Leptospira infection, clearance, persistence, or disease. An in vitro whole blood culture system to study the overall immune response using pathogenic and non-pathogenic Leptospira strains was explored in this study. Using an ELISA, increased IL-8, TNF alpha, and IL-1 in blood samples stimulated with pathogenic and nonpathogenic Leptospira compared to unstimulated controls were detected. In RT2 Profiler PCR Array assays, consistent upregulation of 22 genes and downregulation of 25 genes were observed. Few of the notable upregulated genes included BPI, CCL3, CXCL2, IL-6, IL-8, TLR1, TLR2, TLR6, and TNF and downregulated genes included, LBP, LYZ, MPO, MYD88. IFNβ was upregulated in samples treated with pathogenic Leptospira and IL-1β was upregulated in samples treated with nonpathogenic Leptospira. Toll- like Receptor signaling and expression of pattern recognition receptors were two of the five prominent canonical pathways observed. Individual deconvolution of each of the specific and significant pathways observed in this study may improve the understanding of the pathogenesis of this important zoonotic agent. The use of this system in conjunction with whole transcriptome analysis in a larger population, may unveil the robust nature of host/Leptospira interaction.

Survival time of Leptospira kirschneri on strawberries.

In the past decade, two leptospirosis outbreaks occurred among strawberry harvesters in Germany, with 13, and 45 reported cases respectively. In both outbreaks, common voles (Microtus arvalis) infected with Leptospira kischneri serovar Grippotyphosa were identified as the most likely outbreak source. In an univariate analysis, eating unwashed strawberries was identified as one of the risk factors associated with Leptospira infection. The aim of this study was to evaluate the survival time of L. kirschneri serovar Grippotyphosa on strawberries under varying conditions. Strawberries were spiked with 5x109 of both a laboratory reference strain (strain Moskva V) and an outbreak field strain (94-6/2007) of L. kirschneri serovar Grippotyphosa sequence type 110. Survival times were investigated in a fully crossed design with three incubation times (2h, 4h, 6h and 8h) and three temperatures (15°C, 21°C and 25°C) with three replicated for each condition. A wash protocol was developed and recovered Leptospira were determined by qPCR, dark field microscopy and culturing. Viable L. kirschneri of both the reference strain and the field strain were identified in all samples at 25°C and an incubation time of 2h, but only 1/9 (11%) and 4/9 (44%) of the samples incubated at 15°C were positive, respectively. Both reference and field strain were viable only in 2/9 (22%) at 25° after 6h. After an 8h incubation, viable Leptospira could not be identified on the surface of the strawberries or within the fruit for any of the tested conditions. Based on these results, the exposure risk of consumers to viable Leptospira spp. through the consumption of strawberries bought at the retail level is most likely very low. However, there is a potential risk of Leptospira infection by consumption of strawberries on pick-your-own farms.

Leptospiral protein LIC11334 display an immunogenic peptide KNSMP01

Leptospirosis is considered as a neglected tropical disease which is caused by pathogenic Leptospira spp. The precise mechanisms of leptospirosis pathogenesis are unclear and hence, the progress in development of treatment modalities has been dismal. The present study aimed to identify novel virulent factors of leptospires to understand the disease pathogenesis and to develop treatment modalities. Leptospira interrogans contains two chromosomes and encodes for ~3703 genes, but the functions of several open reading frames have not yet been explored. Among them, novel virulent associated leptospiral proteins (LIC11334, LIC11542, LIC11436, LIC11120 and LIC12539) were identified using VirulentPredict and the antigenicity of these targets was explored by VaxiJen server. Domain architecture of the pathogen specific proteins revealed that LIC11334 had potential to evoke significant immune response against leptospiral infection and LIC11436 contains four folds of immunoglobulin-like domain and plays a vital role in pathogenesis. Therefore, B-cell epitopes were predicted and the epitope of high virulence (and VaxiJen score from LIC11334) was chemically synthesized as peptide (KNSMP01) and labeled with Biotin (Biotin-SGSGEVENPDPKVAQEC). Binding affinity of KNSMP01 with MHC molecules was predicted and the molecule was discovered to have potential to elicit both humoral and cell mediated immune responses and found to interact with host components via hydrophobic interaction, hydrogen bonding and salt bridges. Rabbit antisera was raised against KNSMP01 and found to elicit antigenicity using Western, ELISA and dot blot assays. In silico and in vitro experiments show KNSMP01 to be a promising immunogen and may be a better vaccine candidate for leptospirosis.

Bacteriostatic and Bactericidal Effect of Tigecycline on Leptospira spp.

Tigecycline is a relatively new antimicrobial, belonging to glycylcyclines with antimicrobial activity against a large spectrum of bacteria. Very few data are available on its effect on Leptospira spp., which consist in a bacteriostatic mechanism. The aim of this investigation was to evaluate the bacteriostatic and bactericidal effect of tigecycline on reference Leptospira strains belonging to 16 serovars. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined through the microdilutions method, and tetracycline was used as the control. Results showed that tigecycline had higher MIC and MBC values than tetracycline. Obtained MIC values were between 4 and 32 µg/mL, while MBC values between 16 and >128 µg/mL. Patoc (MIC: 4 µg/mL; MBC: 16 µg/mL) resulted in the most susceptible serovar, while the most resistant were Bataviae (MIC: 32 µg/mL; MBC: 64 µg/mL), Bratislava (MIC: 8 µg/mL; MBC 128 µg/mL), and Tarassovi (MIC: 8 µg/mL; MBC: >128 µg/mL). This is the first investigation focused on the effect of tigecycline against Leptospira spp. reference strains. Since tigecycline is used as a treatment for bacteremia and urinary tract disease, and these symptoms could be linked to Leptospira infection, the possibility of using this antibiotic as a treatment for leptospirosis should be evaluated. Further studies are needed to explore the possibility to use tigecycline for in vivo application against Leptospira.

Leptospira interrogans Retains Direct Virulence After Long Starvation in Water.

Mostly studied as a zoonosis, leptospirosis is also an environment-borne infection and most human cases originate from soil or water contaminations. Yet, only few studies have been interested in the survival of pathogenic Leptospira in freshwater. In this study, water microcosms were designed to evaluate the survival and virulence of Leptospira spp. for 2years. Four commercial bottled drinking waters and a non-ionized water, all previously filter-sterilized, were studied. Either one of two Leptospira interrogans strains, one Leptospira borgpetersenii strain, or a saprophytic Leptospira biflexa was inoculated in these waters under nutrient-deprived conditions. Molecular, microscopic and cultural approaches were used to study Leptospira survival. Direct virulence of the pathogens was assessed using animal challenge without re-culturing. Our results confirmed the capacity of pathogenic Leptospira to survive for more than a year in water. In addition, we showed the ability of L. interrogans in nutrient-deprived conditions to directly cause systemic infection in susceptible animals. To our knowledge, this is the first report of direct infection of a susceptible host with Leptospira following a long starvation and survival period in nutrient-deprived water. Our results also suggest that Leptospira turned into a physiological "survival" state in harsh freshwater conditions. These data are of prime importance considering that freshwater is a major source of Leptospira infections. Environmental survival and virulence of pathogenic Leptospira spp. are becoming a crucial challenge to determine the environmental risk and adopt relevant prevention and control strategies.

MicroRNAs Regulated by the LPS/TLR2 Immune Axis as Bona Fide Biomarkers for Diagnosis of Acute Leptospirosis.

Leptospirosis remains a significant human health issue due to its systemic complications. Therefore, biomarkers that are more effective are urgently needed for the early diagnosis of leptospirosis. MicroRNAs (miRNAs) are evolutionarily conserved regulatory RNAs that have shown the potential to be used as biomarkers for diagnosis, prognosis, and therapy of infectious diseases. In this study, we performed an unbiased screen using the miRNome miRNA array to identify circulating miRNAs with the potential to serve as authentic biomarkers for early diagnosis of leptospirosis. Because leptospiral lipopolysaccharide (LPS) is the predominant leptospiral antigen and plays a vital role in immunological and biological activities, we used LPS treated and untreated in vitro (THP1 cells) and in vivo (BALB/c mice) surrogate models to identify the LPS-specific miRNAs. Differential expression analysis revealed 18 miRNAs to be associated strongly with LPS stimulation in THP1 cells. Of these, three (miR-let-7b-5p, miR-144-3p, and miR-21-5p) were observed to be present at increased levels in vivo The identified miRNAs were validated for their biomarker potential using serum samples from leptospirosis-negative patients and patients with confirmed cases of leptospirosis. Identified miRNAs were able to discriminate the acute leptospiral infection from other febrile diseases with a test sensitivity and specificity of 93.2% and 88.19%, respectively. Gene functional enrichment and protein-protein interaction (PPI) network analysis revealed that the identified miRNAs play important roles in disease signal transduction, signaling by interleukins, the stress-activated protein kinase signaling cascade, the mitogen-activated protein kinase (MAPK) signaling pathway, and the cellular response to a transforming growth factor β (TGF-β) stimulus with a notable interconnection between these biological processes.IMPORTANCE Here, we used miRNAs that are differentially regulated by the LPS/TLR2 immune axis to devise a miRNA-based diagnosis for leptospirosis. The study established the role of the circulating stable miRNAs (miR-21-5p, miR-144-3p, and miR-let-7b-5p) as an early diagnostic marker for leptospirosis. These miRNAs can be used to diagnose acute leptospirosis and also to differentiate leptospiral infection from other bacterial and spirochetal infections, as proved by the use of human clinical samples. Thus, our findings indicate that miRNAs can play a crucial role in the diagnosis of infectious diseases, like leptospirosis, that are generally misdiagnosed.

Early detection of leptospirosis using Anti-LipL32 carbon nanotube immunofluorescence probe

Leptospirosis is a widespread zoonosis and an emerging public health problem. Leptospirosis symptoms are often confused or misdiagnosed with other febrile illness like malaria, viral hepatitis, influenza, dengue, typhoid, melioidosis, and scrub typhus as the clinical manifestations are almost similar. Therefore, early and accurate diagnosis of leptospirosis is indeed critical for proper and prompt treatment. Herein, we report the development of single-walled carbon nanotubes based immunofluorescence probe (Carbo-Lip) for the detection of leptospirosis at an early phase by utilising major outer membrane protein, LipL32 of Leptospira. The Carbo-Lip probe was fabricated through immuno recognition method with fluorescent dye functionalized LipL32 monoclonal antibodies (mAbs), secondary antibody and Leptospira. Surface characterization studies such as Fourier transform infrared spectroscopy with the attenuated total reflectance, scanning electron microscopy, transmission electron microscopy, Zeta potential, and X-ray photoelectron spectroscopy techniques were used to demonstrate the successful fabrication of Carbo-Lip probe. The sensor probe was capable of detecting the presence of leptospires at a lower concentration of 103/ml, and could detect 102 leptospires in 100μL of sample within 3h of the test conditions, and was stable up to 2 weeks. This Carbo-Lip probe was further tested and validated for its capacity to detect Leptospira in clinical samples, which exhibited high selectivity and specificity towards Leptospira even in the presence of malaria and dengue. Our results were consistent with microscopic agglutination test, which is known as gold standard, immunoglobulin M (IgM) enzyme-linked immunoassay (ELISA), IgM spot test, and culture tests for the diagnosis of Leptospira infection.

Serological survey on Leptospirosis of cattle in Cu Chi, Ho Chi Minh City

Leptospirosis is a zoonotic disease of global importance, especially in tropical countries. The purpose of this study to evaluate the seroprevalence of Leptospira in cattle in Ho Chi Minh city. Serum samples from 446 animals were tested by microscopic agglutination test (MAT) using a panel of 24 Leptospira serovars kits provided by Ho Chi Minh City Pasteur Institute. Results showed that seroprevalence of Leptospira at herd-level was 61.54%; and at individual-level was 31.17%. In which, the percentage of beef cattle (29.77%) infected with Leptospira was lower than that of dairy cattle (31.75%). Cattle from 2 to 3 years of age had the highest rate of Leptospira infection in was 37. 72%. According to herd size, the results found that medium-scale farming (from 25 to 50 animals) had the lowest seroprevalence rate with Leptospira spp. (28.65%). No significant differences were found among aged groups of cattle. There were a totally 7 serovars that were detected in cattle, in which, the two most popular serovars were L. Hardjo bovis (31.37%) and L. Hebdomadis (30.26%). There were twelve animals infected with four Leptospira serovars (8.63%); meanwhile, the majority of animals infected with one serovar were 42.45%. The ratio of positive animals at antibody titer of 1:200 was 53.70%. These results indicated a very high exposure of Cu Chi cattle to Leptospira spp. which consequently posed a definite risk for people working with cattle acquiring this zoonotic infection

Acute Kidney Injury and Acute Liver Failure in Leptospira Infection and Weil’s Syndrome

Leptospirosis is considered a zoonosis acquired predominantly from contaminated surfaces and water, more commonly in emerging countries with limited sanitary conditions. Leptospira in the host unleashes an immune response that explains the symptoms and clinical signs; once it reaches the kidney and liver tissue, it can manifest with alterations that lead to acute and chronic diseases in both organs. Weil’s syndrome is the best known clinical manifestation with jaundice and acute kidney injury that could lead to multiple organ failure and death. For its diagnosis, there are simplified scores such as the SPiRO score, the microbiological criteria by microscopy or serological tests; the treatment focuses on antibiotics and, if necessary, provides organic support until the infection is curtailed. The purpose of this review was to address the impact of Lep-tospira infection on the kidney and liver, the mechanisms of organ damage, the clinical presentation, and diagnosis and management of this disease.

Leptospira Infection in African Green Monkeys in an Endemic Area: An Opportunity for Comparative Studies in a Natural Environment.

This study was performed to investigate the potential asymptomatic Leptospira reservoir status among African green monkeys (AGMs) in the Caribbean island of Saint Kitts, and whether there is any renal pathology associated with Leptospira exposure. Forty-eight percent of AGMs tested were positive for Leptospira antibodies by the microscopic agglutination test. Leptospira DNA was detected in 4% of kidney samples tested using a lipl32 gene based PCR. We observed minimal to severe microscopic renal lesions in 85% of the AGM kidneys evaluated. The majority of the AGMs (n = 26) had only minimal to mild interstitial nephritis and a few (n = 3) had moderate to severe lesions. The presence of interstitial nephritis was not significantly associated with Leptospira exposure. The presence of infected AGMs in a small surface limited geographic region may pose zoonotic threat to humans and animals. The impact of Leptospira infection in renal pathology in AGMs warrants further investigation. AGMs residing in a natural setting in an insular, surface limited Leptospira endemic geographic region may offer opportunities for comparative studies to advance the field of leptospirosis. Due to their similarity to humans, such studies in AGMs may also provide translational opportunities to advance Leptospira research.

Meta-analysis of risk factors for canine leptospirosis

Leptospirosis is one of the most widespread zoonotic diseases, with more than one million human cases reported worldwide every year. Dogs could develop infections that range from asymptomatic to severe, and shed leptospires with their urine. Given their close contact with humans, dogs may act both as epidemiological links or as sentinels of pathogenic leptospires in the environment. The aims of our study were to quantitatively summarize the overall prevalence of leptospiral antibodies and to identify factors associated with the probabilities of infection. We searched the electronic databases Scopus, PubMed, PMC and ScienceDirect for observational studies on canine leptospirosis published between 1989 and December 2019 and written in English, Spanish or Portuguese. We fitted a series of multilevel random effects meta-analysis models to estimate the prevalence of antibodies against Leptospira for different types of dogs, health statuses, diagnostic tests, geographic regions and income categories of the countries. We also fitted a number of random effects meta-analysis models to estimate the pooled odds-ratio of factors associated with canine leptospirosis. After removing duplicates and articles not meeting selection criteria, a total of 130 studies in 91 articles were included in this work. We found lower seroprevalence estimates in North America countries (P<0.001) and other high income countries (P<0.001). We also found higher probabilities of leptospiral infection in adult (P=0.017), male dogs with access to the streets (P<0.001). Identifying the profile of dogs that are more exposed to leptospirosis could be useful in the design of public health strategies for the prevention and control of leptospirosis.

Results of seroepidemiological examination of healthy population of Lviv region and representatives of the proffesional risk group for leptospirosis

The objective – a seroepidemiological investigation of population and professional risk group with detection of IgG antibodies to leptospira.Material and methods. Using of enzyme-linked immunosorbent assay (ELISA) for detection of IgG antibodies to leptospira was made. Blood samples of healthy donors and Vodokanal workers were analyzed. Statistical analysis was made by with using of Epitools software.Results. The blood samples of 124 people were examined. 90 samples were collected from healthy donors of Lviv Regional Blood Service Center and 34 samples - from employees of Lviv Vodokanal different units. Among the blood donors, 70 (77.8%) were residents of the regions of Lvivska oblast and 20 (22.2%) residents of Lviv. Among the residents of Lvivska oblast (excluding Lviv city), the number of seropositive persons was 8.57%. The highest percentage of seropositive persons was detected in the Starosambirsky district (20%). Against our expectations, no IgG antibodies were detected in any staff member of Lviv Vodokanal and any residents of Lviv city.Conclusion. The number of people with positive IgG titres to leptospira in the Lviv region is 8.57% (excluding Lviv city), which may indicate past Leptospira infection. The informativeness of ELISA test systems for the detection of anti-leptospirosis IgG antibodies for use in seroepidemiological studies needs further investigation.

Laboratory Diagnosis of Bovine Abortions Caused by Non-Maintenance Pathogenic Leptospira spp.: Necropsy, Serology and Molecular Study Out of a Belgian Experience.

Bovine leptospirosis is a bacterial zoonotic disease caused by pathogenic Leptospira spp. The pathology and epidemiology of this infection are influenced by the numerous existing serovars and their adaptation to specific hosts. Infections by host-maintained serovars such as Hardjo are well documented, unlike those from the incidental ones. In July 2014, an emerging phenomenon of an increased incidence of icteric abortions associated with leptospiral infection occurred in southern Belgium. First-line serological analyses targeting cattle-adapted serovars failed at initial diagnosis. This study provides a comprehensive description of laboratory findings—at the level of necropsy, serology and molecular diagnosis—regarding icteric and non-icteric abortions (n = 116) recorded during this time (years 2014–2015) and associated with incidental infection by serovars such as Grippotyphosa, Australis and Icterohaemorrhagiae. Based on these tests, a diagnostic pathway is proposed for these types of infection in cattle to establish an affordable but accurate diagnosis in the future. These investigations add insights into the understanding of the pathogenesis of bovine leptospirosis associated with serovars classically described as non-maintenance.

Seroprevalence of Leptospirosis among High-Risk Individuals in Morocco.

Background Leptospirosis is an anthropozoonotic reemerging neglected infectious disease underreported in most developing countries. A cross-sectional study was performed between 17 and 23 February 2014 to estimate the seroprevalence of leptospirosis among high-risk populations in Casablanca (Morocco). Methods A total of 490 human serum samples (97.6% males) were collected in 3 high-risk occupational sites including the biggest meat slaughterhouse (n = 208), a poultry market (n = 121), and the fish market (n = 161). A total of 125 human blood samples were also collected from the general population and used in this study as a control group. To detect the presence of anti-Leptospira, sera were screened with in-house IgG and IgM enzyme-linked immunosorbent assay (ELISA). Positive samples were tested by Microscopic Agglutination Technique (MAT) using a panel of 24 serovar cultures and cut point of 1 : 25. Results Seroprevalence of leptospirosis among the control group was 10.4% (13/125). A high seropositivity among the overall seroprevalence of 24.1% (118/490) was observed in the high-risk groups of which 7.3% (36/490), 13.7% (67/490), and 3.1% (15/490) were for anti-Leptospira IgM, IgG, and both IgG and IgM antibodies, respectively. Most of the positive individuals were occupationally involved in poultry (37.2%), followed by the market fish (26.1%) and the meat slaughterhouse (14.9%) workers. Among all ELISA-positive serum samples, 20.3% (n = 24) had positive MAT responses, of which the Icterohaemorrhagiae (n = 7) is the most common infecting serogroup followed by Javanica (4), Australis (2), and Sejroe, Mini, and Panama (one in each). In the remaining 8 MAT-positive sera, MAT showed equal titers against more than one serogroup. Conclusion Individuals engaged in risk activities are often exposed to leptospiral infection. Therefore, control and prevention policies toward these populations are necessary.

New strategies for Leptospira vaccine development based on LPS removal.

Pathogenic spirochetes from genus Leptospira are etiologic agents of leptospirosis. Cellular vaccines against Leptospira infection often elicit mainly response against the LPS antigen of the serovars present in the formulation. There is no suitable protein candidate capable of replacing whole-cell vaccines, thus requiring new approaches on vaccine development to improve leptospirosis prevention. Our goal was to develop a whole-cell vaccine sorovar-independent based on LPS removal and conservation of protein antigens exposure, to evaluate the protective capacity of monovalent or bivalent vaccines against homologous and heterologous virulent Leptospira in hamster. Leptospire were subjected to heat inactivation, or to LPS extraction with butanol and in some cases further inactivation with formaldehyde. Hamsters were immunized and challenged with homologous or heterologous virulent serovars, blood and organs were collected from the survivors for bacterial quantification, chemokine evaluation, and analysis of sera antibody reactivity and cross-reactivity by Western blot. Immunization with either heated or low LPS vaccines with serovar Copenhageni or Canicola resulted in 100% protection of the animals challenged with homologous virulent bacteria. Notably, different from the whole-cell vaccine, the low LPS vaccines produced with serovar Canicola provided only partial protection in heterologous challenge with the virulent Copenhageni serovar. Immunization with bivalent formulation results in 100% protection of immunized animals challenged with virulent serovar Canicola. All vaccines produced were able to eliminate bacteria from the kidney of challenged animals. All the vaccines raised antibodies capable to recognize antigens of serovars not present in the vaccine formulation. Transcripts of IFNγ, CXCL16, CCL5, CXCL10, CXCR6, and CCR5, increased in all immunized animals. Conclusion: Our results showed that bivalent vaccines with reduced LPS may be an interesting strategy for protection against heterologous virulent serovars. Besides the desirable multivalent protection, the low LPS vaccines are specially promising due to the expected lower reatogenicity.