Presence of Leptospira spp. in a Mosaic of Wetlands Used for Livestock Raising under Differing Hydroclimatic Conditions.

Knowledge about the life cycle and survival mechanisms of leptospires in the environment is scarce, particularly regarding the environmental factors associated with their presence in ecosystems subject to livestock farming, where precipitation, seasonal floods, and river overflows could act as facilitators of leptospire dispersion. This study aimed to identify and study the presence of Leptospira spp. in the Lower Delta of the Paraná River and describe the physical, chemical, and hydrometeorological conditions associated with their presence in wetland ecosystems impaired by livestock raising intensification. Here, we show that the presence of Leptospira was determined mainly by water availability. We detected the species Leptospira kmetyi, L. mayottensis, and L. fainei and successfully cultured the saprophytic species L. meyeri from bottom sediment, suggesting the association of leptospires with microbial communities of the sediment's biofilm to enhance its survival and persistence in aquatic environments and adapt to changing environmental conditions. Knowledge of Leptospira sp. diversity in wetlands and the impact of climate variability on the transmission of these organisms is crucial for predicting and preventing leptospirosis outbreaks in the context of human health. IMPORTANCE Wetlands are environments that are often conducive to the survival and transmission of Leptospira because they provide a suitable habitat for the bacteria and are often home to many animal species that can act as reservoirs for leptospirosis. Bringing humans and animals into closer contact with contaminated water and soil and increased frequency and intensity of extreme weather events may further exacerbate the risk of leptospirosis outbreaks, which is mostly relevant in the context of climate change and a widespread intensification of productive activities, particularly in the Lower Delta of the Paraná River. The detection of leptospiral species in wetland ecosystems impaired by livestock raising intensification can help to identify propitious environmental factors and potential sources of infection, develop preventive measures, and plan for appropriate responses to outbreaks, ultimately improving public health outcomes.

Acute phase proteins and total antioxidant capacity in free-roaming cats infected by pathogenic leptospires.

BACKGROUND: Leptospirosis is a neglected but widespread zoonotic disease throughout the world. Most mammals are hosts of Leptospira spp., including domestic cats, species in which no consensus has been reached on the clinical presentation or diagnosis of the disease. The study of acute-phase proteins (APPs) and biomarkers of oxidative status would contribute to knowledge about the disease in cats. This report evaluated four APPs: Serum amyloid A-SAA, Haptoglobin-Hp, albumin and Paraoxonase 1-PON1 and the antioxidant response through Total Antioxidant Capacity-TAC, in 32 free-roaming cats. Cats were classified as seroreactive for anti-leptospiral antibodies (group 1, n = 8), infected with Leptospira spp (group 2, n = 5) and leptospires-free cats (group 3, n = 19). RESULTS: SAA differences were observed between groups 1 and 2 (p-value = 0.01) and between groups 2 and 3 (p-value = 0.0001). Hp concentration differences were only detected between groups 2 and 3 (p-value = 0.001). Albumin concentrations only differed between groups 1 and 3 (p-value = 0.017) and 2 and 3 (p-value < 0.005). Cats in groups 1 (p-value < 0.005) and 2 (p-value < 0.005) had lower PON1 concentrations than group 3. No statistically significant differences between pairs of groups were detected for TAC concentrations. The principal component analysis (PCA) retained two principal components, (PC1 and PC2), explaining 60.1% of the observed variability of the inflammatory proteins and the antioxidant TAC. CONCLUSIONS: Increases in Serum SAA, Hp, and decreases in PON1 activity may indicate an active inflammatory state in infected cats (currently or recently infected).

Risk factors for Leptospira seropositivity in rural Northern Germany, 2019.

We investigated seroprevalence and factors associated with Leptospira spp. infections in humans in rural Northern Germany. Sera of 450 participants were tested for leptospira-reactive IgG antibodies by two enzyme-linked immunosorbent assays (ELISA). A narrow (specific) and a broad (sensitive) case definition were applied and results compared in the analysis. Personal data were collected via questionnaire and associations with the serostatus were investigated by multivariable logistic regression. The seroprevalence estimates were 1.6% (95%-confidence interval (CI) = 0.63-3.2) under the narrow and 4.2% (95%-CI = 2.6-6.5%) under the broad case definition. Few (14%) participants knew about the pathogen. No seropositive participant recalled a prior leptospirosis diagnosis. Spending more than two hours a week in the forest was significantly associated with anti-leptospira IgG in both models (broad case definition: adjusted odds ratio (aOR) = 2.8, 95%-CI = 1.2-9.1; narrow case definition: aOR = 11.1, 95%-CI = 1.3-97.1). Regular cleaning of storage rooms was negatively associated in the broad (aOR = 0.17, 95%-CI = 0.03-0.98) and touching a dead rodent in the past 10 years in the narrow case definition model (aOR = 0.23, 95%-CI = 0.05-1.04). Our findings support risk factors identified in previous investigations. To counter the low awareness for the pathogen, we recommend that health authorities communicate risks and preventive measures to the public by using target-group specific channels.

Survey of selected pathogens in free-ranging pinnipeds in Uruguay.

Marine mammals, regarded as sentinels of aquatic ecosystem health, are exposed to different pathogens and parasites under natural conditions. We surveyed live South American fur seals Arctocephalus australis and South American sea lions Otaria flavescens in Uruguay for Leptospira spp., canine distemper virus (CDV), Mycobacterium spp., Toxoplasma gondii, and Neospora caninum. Samples were collected from 2007 to 2013. The seroprevalence of Leptospira spp. was 37.6% positive, 50.9% negative, and 11.5% suspect for A. australis (n = 61) while for O. flavescens (n = 12) it was 67% positive, 25% negative, and 8% suspect. CDV RNA was not detected in any of the analyzed samples. Most animals tested seropositive to tuberculosis antigens by WiZo ELISA (A. australis: 29/30; O. flavescens: 20/20); reactivity varied with a novel ELISA test (antigens MPB70, MPB83, ESAT6 and MPB59). Seroprevalence against N. caninum and T. gondii was 6.7 and 13.3% positive for O. flavescens and 0 and 2.2% positive for A. australis respectively. To evaluate possible sources of infection for pinnipeds, wild rats Rattus rattus and semi-feral cats Felis catus were also tested for Leptospira spp. and T. gondii respectively. Water samples tested for Leptospira revealed saprofitic L. bioflexa. Pathogenic Leptospira were detected in the kidneys of 2 rats, and cats tested positive for T. gondii (100%). These results represent a substantial contribution to the study of the health status of wild pinnipeds in Uruguay.

Quantitative Real-Time PCR Assays for the Detection of Pathogenic Leptospira Species in Urine and Blood Samples in Canine Vaccine Clinical Studies: a Rapid Alternative to Classical Culture Methods.

Leptospirosis is a vaccine-preventable bacterial zoonotic disease caused by pathogenic Leptospira species. The efficacy of Leptospira canine vaccines is assessed by challenging vaccinated and control dogs with virulent serovars of Leptospira, followed by detection of Leptospira in blood and urine. We assessed the consistency between results obtained for urine and blood samples from clinical studies with species-specific real-time quantitative PCR (qPCR) targeting the lipL32 gene and those obtained with the reference culture method. The specificity of the qPCR assay was confirmed by negative results for nonpathogenic Leptospira and for several canine viruses, bacteria, and parasites. The results from the two methods were compared using McNemar's test, kappa coefficient (κ), and percentage of agreement analyses. The results for numbers of positive and negative dogs were similar, with no false-negative results with the qPCR assay. For both blood and urine, there was strong agreement between the culture method and qPCR results (κ = 0.68 [95% confidence interval (CI), 0.62 to 0.74] and κ = 0.65 [95% CI, 0.59 to 0.71], respectively). However, there was a statistically significant difference between blood samples (P < 0.001) and urine samples (P = 0.028). The negative percentage agreements were 97% and 84% and the positive percentage agreements were 68% and 83% for blood and urine samples, respectively. Although the cell culture method is the recommended gold standard, our results show that qPCR assay is a valid alternative method for the rapid and specific detection of pathogenic Leptospira spp. in urine and blood samples during vaccine efficacy studies, without loss of sensitivity.

Hantavirus-Leptospira coinfections in small mammals from central Germany.

European orthohantaviruses (Puumala orthohantavirus (PUUV); Dobrava-Belgrade orthohantavirus (DOBV), genotype Kurkino; Tula orthohantavirus (TULV)), and Leptospira spp. are small mammal-associated zoonotic pathogens that cause diseases with potentially similar symptoms in humans. We investigated the frequency of Leptospira spp. and hantavirus single and double infections in small mammals from 22 sites in Thuringia, central Germany, during 2017. TULV infections were detected at 18 of 22 sites (mean prevalence 13.8%, 93/674). PUUV infections were detected at four of 22 sites (mean prevalence 1.5%, 7/471), and respective PUUV sequences formed a novel phylogenetic clade, but DOBV infections were not detected at all. Leptospira infections were detected at 21 of 22 sites with the highest overall prevalence in field voles (Microtus agrestis) with 54.5% (6/11) and common voles (Microtus arvalis) with 30.3% (205/676). Leptospira-hantavirus coinfections were found in 6.6% (44/671) of common voles but only in two of 395 bank voles. TULV and Leptospira coinfection probability in common voles was driven by individual (age) and population-level factors. Coinfections seemed to be particularly associated with sites where Leptospira spp. prevalence exceeded 35%. Future investigations should evaluate public health consequences of this strong spatial clustering of coinfections.

First report of pathogenic Leptospira spp. in Tadarida brasiliensis bats (family Molossidae) and Eptesicus furinalis (family Vespertilionidae) of Argentina. New host species in this country?

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

Cross-sectional study of Leptospira spp. in commercial pig farms in the state of Goiás, Brazil.

Leptospirosis is an infectious, contagious disease highly important to the world pig industry, which causes reproductive loss in breeding herds. Endemic infections in a herd may produce little evidence of clinical disease despite resulting in economic losses. However, some epidemiological features of leptospirosis in midwestern Brazil, such as risk factors and prevalence of the disease, remain unclear. Therefore, this study focused on assessing the prevalence of the Leptospira spp. in intensive pig herds and associating its risk factors. A set of 900 blood samples, equally distributed between nursery, growing, and finishing pigs of 30 intensive farrow-to-finish farms, were analyzed using the microagglutination test (MAT), in order to detect anti-Leptospira spp. antibodies for 24 different Leptospira spp. serovars. An occurrence of 4.67% (55/342) seropositive samples were detected in fattening pigs. The variables associated with the disease occurrence were animals per square meter at fattening (OR 0.006, CI 95% 0.004-0.42, p = 0.0105) and pen division between growing and fattening pigs (OR 3.56, CI 95% 0.563-22.541, p = 0.185). Thus, the variables semi-hollow floor in the maternity (OR 16.66; CI 95%: 2.17-128.2 and p = 0.006) and animals per trough at fattening (OR: 0.08, CI 95% 0.009-0.87 and p = 0.025), observed in this study, highlight the importance of the fattening phase in the epidemiology of the disease, bringing information on risk factors involved in the occurrence and dissemination of leptospirosis in intensive pig herds.

New insights on the infection of pathogenic Leptospira species in American mink (Neovison vison) in southern Chile.

Leptospirosis is a zoonosis of global distribution, caused by the infection of pathogenic Leptospira, a group of bacteria capable of infecting both domestic and wild animals. Mink (Neovison vison) in southern Chile is recognized as a wild and synanthropic rodent predator (among various other prey), and Leptospira infection in them can be acquired through contact with the pathogen in the environment or by eating infected prey. Thus, the aim of this study was to provide more specifics regarding the source of the infection for the American mink under the conditions of Southern Chile. Minks were captured in the Los Ríos region, southern Chile, in an area with well-developed dairy farming. Two areas were selected for mink trapping, one with a high degree of dairy farming and a second with a low degree of dairy farming. Within them, 16 study sites were visited, and 45 American mink were trapped and euthanized to obtain kidney tissue and blood serum samples for bacteria isolation and determination of antibodies titers, respectively. Molecular characterization of the isolated strains was performed. Three minks from sites of high-dairy farming industry and only one from sites with low-degree dairy farming were detected as infected through molecular confirmation. This study shows evidence that confirms previous findings made in southern Chile, regarding mink as host of Leptospira interrogans serovar Hardjo-prajitno associated to cattle-farming areas. However, typing information ( Leptospira interrogans Copenhageni and Icterohaemorrhagiae ) suggests that the consumption of rodents may also be a potential source of infection.

Leptospira spp. and Toxoplasma gondii in stranded representatives of wild cetaceans in the Philippines.

BACKGROUND: The stranding events of cetaceans in the Philippines provide opportunities for gathering biological information and specimens, especially from the pelagic forms. As part of an effort to monitor the health of wild cetaceans, this study detected Leptospira spp. and Toxoplasma gondii, causative agents of the emerging zoonotic diseases leptospirosis and toxoplasmosis respectively, in their stranded representatives. From October 2016-August 2018, 40 cetaceans (representing 14 species) that stranded nationwide were sampled for brain, cardiac muscle, skeletal muscle, kidney, and blood tissues, urine, and sera. These were subjected to molecular, serological, culture, and histopathological analyses to detect the target pathogens. RESULTS: T. gondii was detected in 20 (71%) of the 28 cetaceans with biological samples subjected to either molecular detection through RE gene amplification or IgG antibodies detection through agglutination-based serological assay. On the other hand, Leptospira was detected in 18 (64%) of 28 cetaceans with biological samples subjected to bacterial culture, molecular detection through 16S rDNA amplification, or IgM antibodies detection through ELISA-based serological assay. CONCLUSIONS: There is the plausibility of toxoplasmosis and leptospirosis in cetacean populations found in the Philippines, however, acute or chronic phases of infections in sampled stranded individuals cannot be confirmed in the absence of supporting pathological observations and corroborating detection tests. Further studies should look for more evidences of pathogenicity, and explore the specific mechanisms by which pelagic cetacean species become infected by Leptospira spp. and T. gondii. As there is growing evidence on the role of cetaceans as sentinels of land-sea movement of emerging pathogens and the diseases they cause, any opportunity, such as their stranding events, should be maximized to investigate the health of their populations. Moreover, the role of leptospirosis or toxoplasmosis in these stranding events must be considered.

Determination of the seroprevalence of Leptospira spp. and the main serovars circulating in cattle in the province of Manabí, Ecuador. / Determinación de la seroprevalencia de Leptospira spp. y los principales serovares circulantes en el ganado bovino en la provincia de Manabí, Ecuador.

Leptospirosis is a transmissible zoonotic disease caused by pathogenic strains of the genus Leptospira. Clinical signs in cattle are fever, haematuria, haemoglobinuria, meningitis, abortion, birth of weak calves and infertility; in the most severe cases, it can cause death to the animal. The few studies conducted in Ecuador, and in particular the province of Manabí, have revealed varying prevalence rates, ranging from 35.8% to 75%. The objective of this study is to determine the seroprevalence of leptospirosis in cattle and to ascertain the main serovars circulating in the province of Manabí. A cross-sectional epidemiological study was conducted from November 2015 to March 2016, for which seven cantons were selected at random and a total of 854 animals from 67 herds were investigated. The samples were processed in the laboratories of the Animal Diagnostics Directorate of the Ecuadorian Agency for Agriculture Quality Assurance (AGROCALIDAD) in Tumbaco, using the microscopic agglutination test. The sera were analysed to check whether they contained any of the eight serovars of Leptospira interrogans circulating most frequently in the country: Canicola, Hardjo, Pomona, Icterohaemorrhagiae, Grippotyphosa, Wolffi, Bratislava and Copenhageni. Overall seroprevalence at herd level was 97.01%, with the most common serovars being Pomona, Icterohaemorrhagiae, Grippotyphosa, Bratislava and Canicola. It was concluded that there is high seroprevalence at herd level in the province of Manabí.

La leptospirose est une maladie zoonotique infecto-contagieuse causée par des souches pathogènes du genre Leptospira. Chez les bovins, le tableau clinique de la maladie se caractérise par de la fièvre, une hématurie, une hémoglobinurie, une méningite, des cas d'avortement ou la naissance de veaux faibles et une infertilité ; dans les cas les plus graves, la maladie peut provoquer la mort de l'animal. En Équateur et plus précisément dans la province de Manabí, les rares études effectuées jusqu'à ce jour ont rapporté des taux de prévalence allant de 35,8 % à 75 %. Les auteurs présentent les résultats d'une étude visant à déterminer la prévalence sérologique de la leptospirose dans le cheptel bovin de la province de Manabí ainsi que les principaux sérovars présents. Une enquête épidémiologique transversale a été conduite de novembre 2015 à mars 2016 dans sept cantons choisis de manière aléatoire. Au total, 854 animaux appartenant à 67 élevages ont été testés. Les prélèvement ont été préparés par les laboratoires de la Direction du diagnostic vétérinaire de l'Agence équatorienne de la qualité agricole (AGROCALIDAD) de Tumbaco et soumis à une épreuve d'agglutination microscopique afin de déterminer s'ils contenaient l'un ou plusieurs des huit sérovars de Leptospira interrogans présents dans le pays, à savoir Canicola, Hardjo, Pomona, Icterohaemorrhagiae, Grippotyphosa, Wolffi, Bratislava et Copenhageni. La séroprévalence globale au niveau des troupeaux était de 97,01 %, les sérovars les plus fréquents étant Pomona, Icterohaemorrhagiae, Grippotyphosa, Bratislava et Canicola. Les auteurs en concluent que la séroprévalence au niveau des troupeaux est élevée dans la province de Manabí.

La leptospirosis es una enfermedad zoonótica infectocontagiosa causada por cepas patógenas del género Leptospira. En el ganado bovino, clínicamente se caracteriza por fiebre, hematuria, hemoglobinuria, meningitis, abortos y/o nacimiento de animales débiles e infertilidad, y, en los casos más graves, puede provocar la muerte de los animales. En Ecuador, y en concreto en la provincia de Manabí, los escasos estudios realizados denotan prevalencias variables, que van del 35,8% al 75%. El objetivo del presente estudio es determinar la seroprevalencia de la leptospirosis en el ganado bovino y conocer los principales serovares circulantes en la provincia de Manabí. Se realizó un estudio epidemiológico de tipo transversal en el período comprendido entre noviembre de 2015 y marzo de 2016, para el cual se seleccionaron siete cantones al azar y se investigaron un total 854 animales pertenecientes a 67 hatos. Las muestras se procesaron en los Laboratorios de la Dirección de Diagnóstico Animal de la Agencia Ecuatoriana para la Calidad del Agro (AGROCALIDAD) de Tumbaco utilizando la técnica de aglutinación microscópica, y los sueros se analizaron para comprobar si contenían cada uno de los ocho serovares de Leptospira interrogans de mayor circulación en el país: Canicola, Hardjo, Pomona, Icterohaemorrhagiae, Grippotyphosa, Wolffi, Bratislava y Copenhageni. La seroprevalencia general a nivel de hatos fue del 97,01%, siendo los más frecuentes los serovares Pomona, Icterohaemorrhagiae, Grippotyphosa, Bratislava y Canicola. Se llegó a la conclusión de que en la provincia de Manabí existe una alta seroprevalencia a nivel de hatos.

Pharmacokinetics of enrofloxacin HCl-2H2 O (ENRO-C), PK/PD, and Monte Carlo modeling vs. Leptospira spp. in cows.

The pharmacokinetics, PK/PD ratios, and Monte Carlo modeling of enrofloxacin HCl-2H2 O (Enro-C) and its reference preparation (Enro-R) were determined in cows. Fifty-four Jersey cows were randomly assigned to six groups receiving a single IM dose of 10, 15, or 20 mg/kg of Enro-C (Enro-C10 , Enro-C15 , Enro-C20 ) or Enro-R. Serial serum samples were collected and enrofloxacin concentrations quantified. A composite set of minimum inhibitory concentrations (MIC) of Leptospira spp. was utilized to calculate PK/PD ratios: maximum serum concentration/MIC (Cmax /MIC90 ) and area under the serum vs. time concentration of enrofloxacin/MIC (AUC0-24 /MIC90 ). Monte Carlo simulations targeted Cmax /MIC = 10 and AUC0-24 /MIC = 125. Mean Cmax obtained were 6.17 and 2.46 µg/ml; 8.75 and 3.54 µg/ml; and 13.89 and 4.25 µg/ml, respectively for Enro-C and Enro-R. Cmax /MIC90 ratios were 6.17 and 2.46, 8.75 and 3.54, and 13.89 and 4.25 for Enro-C and Enro-R, respectively. Monte Carlo simulations based on Cmax /MIC90  = 10 indicate that only Enro-C15 and Enro-C20 may be useful to treat leptospirosis in cows, predicting a success rate ≥95% when MIC50  = 0.5 µg/ml, and ≥80% when MIC90  = 1.0 µg/ml. Although Enro-C15 and Enro-C20 may be useful to treat leptospirosis in cattle, clinical trials are necessary to confirm this proposal.

The prevalence of pathogenic forms of Leptospira in natural populations of small wild mammals in Serbia.

The greatest epidemiological significance of leptospirosis in Europe comes from the fact that it is the most widespread zoonosis in the world. However, epizootiological data, especially information on maintenance hosts such as small wild mammals, are largely missing. To fill this gap in data in Serbia, we used RT-PCR for the detection of pathogenic Leptospira species and analysed 107 animals belonging to six species of small wild mammals (Apodemus agrarius, Apodemus flavicollis, Microtus arvalis, Myodes glareolus, Microtus subterraneus and Sorex araneus) collected from two localities. The animals from the first locality that was situated in a tourist area, were collected for four consecutive years (2014-2017). We found persistent incidence of infection from year to year ranging from 6.67% to 78.57%. The average frequency of infected animals was 33.3% with the highest frequency in 2014, the year characterised by a very high number of flooding days. All animals proved to be infected with pathogenic Leptospira species that were collected from the second locality situated in an agricultural area in a single year, 2014. The findings show a variable but constant presence of pathogenic Leptospira species in populations of small wild mammals in the studied areas, which indicates the need for constant monitoring.

Isolation of saprophytic Leptospira spp. from a selected environmental water source of Argentina.

Ten Leptospira spp. strains were isolated from water samples from Nievas stream, Olavarría, Buenos Aires province (Argentina). The isolates showed the typical motility and morphology of the genus Leptospira under dark field microscopy, developing in liquid EMJH medium after eight days of incubation at 13°C and 30°C. All isolates were negative by the Multiple Locus Variable Number Tandem Repeat Analysis (MLVA). Molecular identification by 16S rRNA gene sequencing identified all isolates as nonpathogenic leptospires. Four isolates showed a genetic profile identical to that of the reference strain Leptospira biflexa serovar Patoc, and six isolates revealed sequence similarities within the 97-98% range, closely related to Leptospira yanagawae and Leptospira meyeri, respectively. Strains ScialfaASA42, ScialfaASA45, ScialfaASA44, ScialfaASA47, ScialfaASA49, ScialfaASA50 and ScialfaASA51 possibly represent a novel species of the genus Leptospira.

[Differentiation of pathogenic leptospires spp by PCR of ligB gene and sequencing]. / Diferenciación de serovares de leptospiras patógenas mediante PCR del gen ligB y secuenciación.

Leptospirosis is a zoonosis having worldwide distribution. The objective of this work was to develop a molecular technique to differentiate pathogenic Leptospira spp. A region of adhesin ligB, present only in the pathogenic species was amplified by PCR and sequenced. ligBRpet and ligBFpet primers were used, which amplified the target DNA from pathogenic L. interrogans reference strains serovars Pomona strain Pomona, Canicola strain Hond Utrecht IV, Copenhageni strain M 20, Wolffi strain 3705, Pyrogenes strain Salinem, Hardjo strain Hardjoprajitmo, L. borgpetersenii serovar Castellonis strain Castellon 3 and 4 pathogenic strains isolated from bovines, pigs, rats and opossums. L. biflexa serovars Patoc strain Patoc I and Andamana strain Andamana were not amplified. Sequencing of the amplified products exhibited sufficient variation among serovars, which differentiates them.

Leptospirosis in Poland in the years 2014-2017 ­ characteristics of infections and epidemiological surveillance data

OBJECTIVES: The aim of the study was the analysis of current Leptospira spp. infections in Poland on the basis of blood serum samples tests results and clinical data collected from clinicians in the Laboratory NIPH-NIH. METHODS: Clinical materials from 48 patients with clinical symptoms suggesting Leptospira spp suspected of leptospirosis from the years 2014-2017 were included to the study. Blood serum samples collected from patients were tested in Laboratory of Rickettsiae, Chlamydiae and Spirochaetes (currently Laboratory of Vector-borne Diseases) of NIPH-NIH. Levels of specific IgM and IgG antibodies to Leptospira spp. antigens were detected with the enzyme-linked immunosorbent assay (ELISA). RESULTS: Specific antibodies to Leptospira spp. were detected in 18 patients (37.5%). IgM antibodies were found in 6 patients (12.5%) and IgG antibodies were identified in 7 patients (14.6%). Both classes of antibodies of were detected in 5 patients (10.4%). The most samples for study were sent to laboratory from Masovian (13 samples) and Kuyavian-Pomeranian (11 samples) Voivodeships. Not any samples from the Lower Silesia, Lublin, Lódz, Podlaskie and Warmian-Masurian Voivodeships were received. In these patients the most common symptoms of disease were: fever, hepatitis with jaundice and renal failure. CONCLUSIONS: The number of diagnosed human leptospirosis in Poland is low in comparison to the number of cases in other countries, although the Leptospira spp. spirochetes occur in animals in the environment.

Neglected zoonotic agents in cattle abortion: tackling the difficult to grow bacteria.

BACKGROUND: Coxiella burnetii, Chlamydia abortus and Leptospira spp. are difficult to grow bacteria that play a role in bovine abortion, but their diagnosis is hampered by their obligate intracellular lifestyle (C. burnetii, C. abortus) or their lability (Leptospira spp.). Their importance is based on the contagious spread in food-producing animals, but also as zoonotic agents. In Switzerland, first-line routine bacteriological diagnostics in cattle abortions is regulated by national law and includes only basic screening by staining for C. burnetii due to the high costs associated with extended spectrum analysis. The aim of this study was to assess the true occurrence of these zoonotic pathogens in 249 cases of bovine abortion in Switzerland by serology (ELISA for anti-C. burnetii and C. abortus antibodies and microscopic agglutination test for anti-Leptospira spp. antibodies), molecular methods (real-time PCR and sequencing of PCR products of Chlamydiales-positive cases), Stamp's modification of the Ziehl-Neelsen (mod-ZN) stain and, upon availability of material, by histology and immunohistochemistry (IHC). RESULTS: After seroanalysis the prevalence was 15.9% for C. burnetii, 38.5% for C. abortus and 21.4% for Leptospira spp. By real-time PCR 12.1% and 16.9% of the cases were positive for C. burnetii and Chlamydiales, respectively, but only 2.4% were positive for C. burnetii or Chlamydiales by mod-ZN stain. Sequencing of PCR products of Chlamydiales-positive cases revealed C. abortus in 10% of cases and the presence of a mix of Chlamydiales-related bacteria in 5.2% of cases. Pathogenic Leptospira spp. were detected in 5.6% of cases. Inflammatory lesions were present histologically in all available samples which were real-time PCR-positive for Chlamydiales and Leptospira spp. One of 12 real-time PCR-positive cases for C. burnetii was devoid of histological lesions. None of the pathogens could be detected by IHC. CONCLUSION: Molecular detection by real-time PCR complemented by histopathological analysis is recommended to improve definitive diagnosis of bovine abortion cases and determine a more accurate prevalence of these zoonotic pathogens.

Prevalence of renal lesions in slaughtered cattle in Shiraz, Iran, and detection of Leptospira in them by nested PCR-RFLP.

Renal diseases in cattle are frequently not recognized due to the subclinical conditions. Some species of Leptospira are the main cause of infectious agents that damage the kidneys and lead to abortion and economic losses in cattle and are also of major concern in the public health. This study was aimed to assess the prevalence of renal lesions of slaughtered cattle in the Shiraz abattoir and to determine the correlation between rejected kidneys and infection with Leptospira using nested PCR-restriction fragment length polymorphism (RFLP) techniques. Out of 1000 inspected animals, 205 (20.5 %) revealed the renal lesions. Chronic nephritis (7.5 %), white-spotted kidney (7.3 %), and petechial hemorrhage (3.5 %) were the most prevalent forms of the lesions. A direct correlation between increasing the age and significant increase in the rate of lesions was also observed (P = 0.03). Using nested PCR-RFLP assay, 40.8 % of the tested kidneys were turned to be infected to the pathogenic species of Leptospira. The risk of infection of the kidneys with white spot to pathogenic species of Leptospira (53.8 %) was more than that of the kidneys with other lesions (25.0 %) (P = 0.014). The odd ratio indicates that the kidneys with white spot lesions are likely to be infected with pathogenic species of Leptospira, five times greater than other lesions. This study showed that renal lesions especially white-spotted kidney, which were considerably associated with Leptospira in slaughtered cattle in Shiraz, were very high. This is important in terms of public health and in particular, increases the risk of transmission of disease to human specially in the high-risk careers including farmers, veterinarians, and abattoir workers.

High Prevalence of Intermediate Leptospira spp. DNA in Febrile Humans from Urban and Rural Ecuador.

Leptospira spp., which comprise 3 clusters (pathogenic, saprophytic, and intermediate) that vary in pathogenicity, infect >1 million persons worldwide each year. The disease burden of the intermediate leptospires is unclear. To increase knowledge of this cluster, we used new molecular approaches to characterize Leptospira spp. in 464 samples from febrile patients in rural, semiurban, and urban communities in Ecuador; in 20 samples from nonfebrile persons in the rural community; and in 206 samples from animals in the semiurban community. We observed a higher percentage of leptospiral DNA-positive samples from febrile persons in rural (64%) versus urban (21%) and semiurban (25%) communities; no leptospires were detected in nonfebrile persons. The percentage of intermediate cluster strains in humans (96%) was higher than that of pathogenic cluster strains (4%); strains in animal samples belonged to intermediate (49%) and pathogenic (51%) clusters. Intermediate cluster strains may be causing a substantial amount of fever in coastal Ecuador.

Sero-epidemiological survey for brucellosis, leptospirosis, and toxoplasmosis in free-ranging Alouatta caraya and Callithrix penicillata from São Paulo State, Brazil.

BACKGROUND AND METHODS: Sera were tested for Brucella spp., Leptospira spp. and Toxoplasma gondii antibodies in 68 free-ranging New World monkeys from a forest fragment of the Brazilian Cerrado. RESULTS AND CONCLUSION: All animals were negative for Brucella spp. and Leptospira spp. However, 75% of Alouatta caraya and 16.6% of Callithrix penicillata were positive for T. gondii. The implications for conservation and health management are discussed.