Leptospira Sp Research Articles

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.

KAJIAN LOKASI TERCEMAR LEPTOSPIRA Sp PADA KASUS FOKUS DI RW 02 KELURAHAN BANDARHARJO KOTA SEMARANG TAHUN 2019

Data in RW 02 Bandarharjo Semarang regency shows that every year since 2017-2019 there are always deaths due to leptospirosis. The purpose of this study is to mapping of leptospirosis and the mount of Rattus sp infected with Leptospira sp, describe the environmental conditions and hygiene behavior. A cross sectional method was used in this study. The study population included Rattus sp. and all houses in RW 02 Bandarharjo. Rattus sp. samples were all captured at the study site, while home samples totaled 48 houses around the house of leptospirosis patients within a radius of 150 meters. Data collection is done through environmental observations, interviews, collecting coordinate points, trapping, and laboratory tests. Examination of rat kidney blood samples was done by gram negative staining. A total of 13 rats were caught Rattus sp, 15.4% of them were indicated to be strongly positive containing Leptospira sp. All conditions of landfills are in an open condition, 89.6% of them are in front of the house, 45.8% of sewers are flooded with water that does not flow. There are 20.8% of respondents do not have the habit of washing hands / feet with soap, more than 50% of respondents ignore the use of protective equipment when cleaning sewers, warehouses, and rat droppings. There are 2 Rattus sp which are indicated to be strongly positive containing Leptospira sp. so that the mobility of Rattus sp can spread leptospirosis at the location in the buffer region. Keywords: leptospirosis, mapping, environment, health behaviour

Use of serological and molecular techniques for detection of Leptospira sp. carrier sheep under semiarid conditions and the importance of genital transmission route

It is possible that there are peculiarities in the epidemiology of leptospirosis in regions with a semiarid climate, where the environment is often adverse, allowing the occurrence of alternative transmission routes. The objective of the work was to generate contributions to the diagnosis and epidemiology of Leptospira sp. infection in sheep reared in semiarid conditions, using serological, molecular and microbiological techniques for diagnosis in dry and rainy seasons. Samples of blood, vaginal fluid, urine, bladder, kidney, uterus, uterine tube, ovary and placenta were collected from 104 sheep (52 animals per season – dry and rainy) slaughtered in the Brazilian semiarid. Diagnostic tests performed were microscopic agglutination test (MAT), polymerase chain reaction (PCR) and bacterial isolation. Anti-Leptospira sp. antibodies were found in 26 (25%) of the animals analyzed by MAT at 1:50 dilution (cut-off 50), while 69 (66.3%) animals had at least one organ/fluid with the presence of Leptospira sp. DNA. Overall, PCR was performed on 758 fragments of organs/fluids from the genital and urinary tracts, and 519 (68.5%) samples tested positive. PCR-positivity was statistically different in dry (46.2%) and rainy (11.5%) seasons for vaginal fluid. It was possible to perform the DNA sequencing in nine samples with 99% similarity to L. interrogans and recovery of viable strains in three samples of vaginal fluid. Regardless of the biological material used in PCR to detect carrier animals and the season, the highest MAT sensitivity values were obtained with cut-off 50 compared to 100. The results obtained indicate that, even in the adverse environmental conditions of the semiarid region, leptospires may survive and propagate by alternative routes of transmission, such as sexual, and the presence of PCR-positive genital tracts in ewes suggests that sexual transmission may play an important role in the epidemiology of the disease in sheep in Brazilian semiarid. In addition, it is suggested the use of titer 50 as cut-off point at serology in semiarid conditions.

Leptospira interrogans infection in tegu lizard (Tupinambis merianae), Brazil

ABSTRACT: We used 12 tegu lizards (Tupinambis merianae) from northeastern Brazil, and we reported nine (75%) Leptospira sp. PCR-positive animals and six (50%) seropositive. Leptospira sp. DNA sequencing revealed 99% similarity with L. interrogans. Our findings indicated that this species may play a role in the transmission of human leptospirosis.

Influence of breed on the clinical and hemato-biochemical parameters in sheep experimentally infected with Leptospira sp.

Early diagnosis of leptospirosis may aid in a favorable prognosis in infected animals, but there are few reports of clinical and hematochemical changes in the ovine species, nor whether the breed exerts any influence on the response to infection. Thus, the objective of the present study was to evaluate the clinical, biochemical and hematological alterations in Santa Inês and crossbred ewes challenged with Leptospira interrogans serogroup Pomona serovar Kennewicki. Twenty-four sheep were used in this experiment, 12 crossbred (group A) and 12 Santa Inês (group B). In each group, sheep were conjunctivally and intraperitoneally challenged. During 60 days post-infection the ewes were evaluated for the presentation of clinical signs and the blood was collected for hemogram and serum biochemistry. Concentration of urea and creatinine; serum aspartate aminotransferase activity (AST); gamma-glutamyltransferase (GGT); total protein and albumin; total bilirubin, direct and indirect were analyzed. The urine of these sheep was collected for urinalysis. Only two Santa Inês sheep showed blood in the urine. Clinical signs implicated in Leptospira sp. infection were not identified. Some sheep had anemia, especially crossbred. However, anemia may be attributed to the more effective cellular response that has been identified in crossbred animals. Only one animal presented leukocytosis with neutrophilia, while 11 presented atypical leukopenia, especially those of the Santa Inês breed. There was a decrease in total protein and albumin, as well as the increase in gammaglutamyltranspeptidase (GGT), especially in Santa Inês sheep. The level of aspartate aminotransferase (AST) remained within the normal range for the species. A significant increase (p < 0.05) in conjugated bilirubin levels in challenged animals was detected. Only one sheep intraperitoneally challenged presented a high level of urea in the blood, but the creatinine level remained within the normal range. The intraperitoneal route was responsible for more significant changes (p < 0.05) in the hemogram and biochemistry when compared to the conjunctival route. The results indicate that crossbred sheep have a more efficient cellular response than Santa Inês sheep, which may confer a greater resistance to infection. Clinical signs are not good parameters to follow the development of leptospirosis in crossbred and Santa Inês breed. Hematological and biochemical analyzes were useful in the detection of anemia and possible liver changes caused by leptospirosis. The intraperitoneal route was able to cause more conclusive alterations of the analyzed parameters, however, it is possible that the alterations caused by the conjunctival route reproduce in a more faithful way what happens in a natural situation of infection.

LeptospiraSeroprevalence Detection and Rabies Virus Absence in an Urban Raccoon (Procyon lotor) Population in a Highly Populated Area, Costa Rica

Leptospirosis and rabies are zoonotic diseases of public health importance and endemic diseases in tropical countries such as Costa Rica. Peridomestic wild animals such as raccoons (Procyon lotor) have been implicated as competent hosts of Leptospira spirochetes and rabies virus. This study focused on understanding the role of urban raccoons in the dynamics of leptospirosis and rabies in a tropical environment. A total of 97 specimens of the common raccoon were captured within the Greater Metropolitan Area of Costa Rica; 32.6% (31/95) of raccoons presented evidence of antibodies (> 1: 100) against Leptospira sp. Attempts to cultivate Leptospira failed, but 19 serovars were identified, which are also responsible for causing disease in humans in Costa Rica. Detected titers ranged from 1: 100 to 1: 6400. Lymphoid hyperplasia in kidneys and spirochetes were demonstrated in 3 of 20 necropsied cases (15%). Twenty brain samples were analyzed by hematoxylin and eosin stain for evidence of encephalitis and Negri body detection and simultaneously frozen brain material was employed to perform a rapid immunoassay test for rabies antigen. All tested samples were negative. This study is the first report of Leptospira seroprevalence in raccoons that cohabit urban areas in Costa Rica. We also highlight the importance of the raccoon as one of their natural competent host and sentinel animals within highly populated urban environments in tropical cities.

Leptospirosis Occurrence in Agricultural Communities in Setiu, Terengganu

Leptospirosis is a re-emerging infectious disease of public health importance. Infection can occur through occupational and environmental exposure of the causative agent through contaminated water in the agricultural field. In this study, soil and water samples were screened for the presence of Leptospira sp. In addition, the study aimed to explore the knowledge, attitudes and practice (KAP) on leptospirosis among agricultural communities in Setiu, Terengganu. A total of 40 environmental samples were randomly collected from rice fields, rubber plantation and oil palm plantation. All samples were examined for the presence of Leptospira sp by quantitative Real Time Polymerase Chain Reaction (qPCR) assay. KAP survey was carried out among 248 respondents using standardized modified translated questionnaires. Findings from this study revealed that only 5% of water collected from rice field area showed the presence of Leptospira sp. Similarly, only 5% of soil collected from oil palm plantation showed the presence of Leptospira sp DNA. Majority of respondents (52.58%) had moderate knowledge and unsatisfactory practice score (72.51%) but showed 75.21% of satisfactory attitude score. The findings also showed there is a significant difference in knowledge with regards to the signs and symptoms of leptospirosis (0.004). Furthermore, the practice score on rat control measures estimated 0.011 between three groups of respondents. The presence of Leptospira sp in the agricultural area and medium knowledge and practice among agricultural communities possesses environmental risks of contracting to leptospirosis. Hence, necessary interventions should be implemented by relevant authorities to prevent future transmission of leptospirosis in agricultural areas

Plasma proteome profiling reveals differentially expressed lipopolysaccharide-binding protein among leptospirosis patients.

Human leptospirosis, or commonly known as "rat urine disease" is a zoonotic disease that is caused by the bacteria called Leptospira sp. The incidence rate of leptospirosis has been under-reported due to its unspecific clinical symptoms and the limitations of current laboratory diagnostic methods. Leptospirosis can be effectively treated with antibiotics in the early stage, and it is a curable disease but the accuracy to diagnose the infection is rarely achieved. The present pilot study investigated plasma protein profiles of leptospirosis patients and compared them against two control groups which consisted of dengue patients and healthy individuals. The plasma protein digests were analyzed using shotgun approach by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Protein abundances were estimated from the exponentially modified protein abundance index (emPAI) values. Plasma proteins in leptospirosis patients with at least two-fold differential expression compared to dengue and healthy control groups (p<0.05, ANOVA) were identified. Lipopolysaccharide (LPS)-binding protein (LBP) was found to be the only protein that has significant different expression between leptospirosis and the two control groups. The expression levels of leucine-rich alpha-2-glycoprotein (LRG1) and alpha-1-antichymotrypsin (ACT) were different significantly between leptospirosis and healthy group but not to the dengue control group. This is the first plasma proteome-based study on leptospirosis that reports the differential expression of LBP compared to both dengue and healthy controls, which has not been previously reported in the context of leptospirosis.

IDENTIFIKASI DNA LEPTOSPIRA SP PADA SAMPEL AIR DAN TANAH DI KOTA MAKASSAR

Leptospirosis is a zoonotic disease that can be found all over the world, caused by a pathogenic leptospira genus. This study aims to identify the presence of Leptospira bacterial DNA in water and soil at several locations in Makassar. This research was conducted in microbiology laboratory Faculty of Medicine Hasanuddin University. A total of 16 samples were taken consisting of water and soil samples taken from four locations in Makassar City namely eggplant market, slums, TPA (final landfill) tamangappa and slaughterhouse (RPH). Samples were then tested using the nested RT-PCR method . The results of this study showed that from 16 samples tested showed one sample with positive to the presence of Leptospira sp bacteria obtained from the soil found in slums located in makassar.

High frequency of genital carriers of Leptospira sp. in sheep slaughtered in the semi-arid region of northeastern Brazil.

Although some studies in sheep have indicated leptospire colonization of the genital tract, further studies are needed to clarify the role of genital carriers in this species. Thus, this study aimed to evaluate the colonization of pathogenic leptospires in the genital and urinary tract of slaughtered sheep. Fifty-seven adult, female woolless sheep destined for slaughter were used. Renal (n = 57), bladder (n = 57), ovary (n = 34), uterine tube (n = 44), and uterus (n = 33) samples were collected for molecular detection of Leptospira sp. DNA, and blood samples (n = 57) for serological testing. The molecular testing was performed using polymerase chain reaction (PCR), and the serological testing was performed using microscopic serum agglutination test (MAT). Samples with amplifying DNA were subjected to genetic sequencing. In total, leptospiral DNA was found in the tissues of 44 (77.2%) sheep, whereas only nine animals were positive on both PCR and MAT; there was slight agreement between PCR and MAT techniques (k = 0.0268; p = 0.684). In 61 (54.9%) genital tract and in five (4.4%) urinary tract samples, the leptospiral DNA was detected, with significant difference (p < 0.001). The genes of one sample from the uterine tube and another from the bladder were sequenced and demonstrated 99% similarity to Leptospira interrogans. Anti-Leptospira antibodies were detected in 11 (19.3%) of the tested animals. The results reinforce the importance of the genital tract as an extra-renal site of colonization, suggesting the possibility of venereal transmission in sheep.

Strategies of the control of an outbreak of leptospiral infection in dairy cattle in Northeastern Brazil.

The aim of the present study was to describe the strategies of the control of an outbreak of leptospiral infection in dairy cattle in Maranhão State, Northeastern Brazil. In the period from January to July 2015, 18 (17%) out of 106 cows presented abortion, six (5.7%) stillbirth, and 12 (11.3%) repeated estrus, totaling 24 animals with reproductive problems. The diagnosis of leptospirosis was based on serology (microscopic agglutination test-MAT), bacteriological culture, and polymerase chain reaction (PCR). Antibiotic therapy, vaccination protocols, and changes in management practices were suggested as control measures. Of all animals on the farm (n = 280), 136 (48.6%) were seropositive for at least one serovar of Leptospira sp. No pure leptospiral culture was obtained. Eight of the animals with reproductive problems yielded positive PCR results (vaginal fluid of seven animals and urine and vaginal fluid of one animal). Genetic sequencing of a vaginal fluid/urine PCR-positive sample revealed Leptospira borgpetersenii. One year after the adoption of control measures, no reproductive problems were observed. Thus, leptospirosis probably caused the reproductive failures in the herd, and the control and prevention measures implemented were efficient in controlling the disease.

Susceptibility among breeds of sheep experimentally infected with Leptospira interrogans Pomona serogroup

Leptospirosis is a disease that negatively affects the productive and reproductive indices of ruminants. Sheep are considered highly resistant to infection, although susceptibility may vary among breeds. Thus, the aim of the present study was to analyze the susceptibility between sheep breeds to the experimental infection by leptospires of the Pomona serogroup. Pomona serogroup, Kennewicki serovar strain (1 × 107 bacteria) was inoculated via the conjunctival route in 12 sheep divided into two groups, one comprising Santa Inês ewes and the other comprising crossbred sheep. In each group, five ewes were challenged with the bacterial strain and one was used as control. All sheep were monitored for 60 days, during which blood samples were collected for serological diagnosis and urine and vaginal fluid samples for molecular and microbiological analyses. Finally, as ewes were submitted to euthanasia and necropsy, some tissues of interest were collected for microbiological, molecular, and histopathological diagnoses. The groups were compared regarding the number of positive reactions according to diagnostic tests. All sheep in each group presented antibodies to Leptospira in all serological analyses, except animals of the control group. However the Santa Inês sheep presented higher concentration and duration of the titers, and their positive reactions were detected earlier than those in crossbred sheep. The antibody titers in group A (median 200, geometric mean 317.48) were significantly different from the group B (median 800, geometric mean 918.96) at D60 post-infection (P = 0,032). The Santa Inês sheep presented a higher number of positive reactions than did the crossbred sheep in the molecular diagnostic tests. According to the molecular diagnosis, the Santa Inês sheep presented more reactions (urine and vaginal fluid) compared to crossbred ewes, but there was no predominance in the detection of leptospiral DNA when comparing urine and vaginal fluid results, nor even between the number of positive kidneys and uterus. The Santa Inês sheep presented a higher number of positive bacteriological cultures. No sheep in either group presented alterations in anatomopathological and histopathological findings. Pure-bred sheep may be more susceptible than crossbred sheep to infection by Leptospira sp. Our findings emphasize the importance of the genital tract as a site of extraurinary infection and indicate the possibility of venereal transmission in the species.

The Presence Of Rat And House Sanitation Associated With Leptospira sp. Bacterial Infection In Rats (A Cross Sectional Study In Semarang, Central Java Province, Indonesia)

The Gajah Mungkur sub-district in Semarang, Indonesia had highest leptospirosis cases (reported in human with seven infected and one dead) in 2015. The purpose of this study was to analyze the association between house sanitation and density of rats with Leptospira sp. infection in rats. The study design was cross sectional observational analytic. The number of 308 trapswere placed in study sites over three consecutive nights afterwards. Every houses were placed with four traps, inside and outside. Trapped rats were anesthetized with atropine dose from 0.02 to 0.05 mg/kg body weight of rats continued with Ketamine HCL dose of 50-100 mg/kg body weight of rats by injecting in the thick thigh muscle of it. After that, identification of rats by species and gender then continues with surgery in which a kidney sample was taken to confirm the presence of bacteria Leptospirasp using PCR techniques. The trap installed in 77 houses which later had further observation on house sanitation which includes the existence of a pile of used goods, food storage, garbage can, and the presence of the ceiling, windows and other ventilation.Data was analyzed using distribution frequency and bivariate chi-square test. We had 100 rats captured with live traps as the samples.The proportion of Rattusnorvegicuswas 27% (14.8% positive Leptospira sp.infection) and Rattustanezumi 73% (11%positive Leptospira sp.infection). The proportion of male and female rats were almost equal. The statistic test result was significant between the density of rats (p = 0.0001, OR 12.833, 95%CI: 1.565-105.261) and sex of rats (p = 0.019, OR 0.095, 95%CI: 0.012–0.769) with Leptospira sp. infection in rats. The number of rats may increase the infection of Leptospirasp., especially female rats and poor condition of house sanitation. It is recommended to improve house sanitation and regularly trapping rats.

Reproductive Disorders and Leptospirosis: A Case Study in a Mixed-Species Farm (Cattle and Swine).

Animal leptospirosis, exempt in rodents, manifests as peculiar biology where the animal can function, simultaneously or not, as a susceptible host or reservoir. In the first case, clinical symptoms are likely. In the second case, infection is subclinical and manifestations are mild or absent. Mild clinical symptoms encompass reproductive failure in production animals for host-adapted Leptospira sp. serovars. This work presents a study on Leptospira sp. infection in a mixed-species (bovine and swine) farm with documented reproductive disorders in the cattle unit. A long calving interval (above 450 days) was the hallmark observed in cows. Some cows (2/26 tested) presented a high titre of antibodies against Leptospira sp. serogroup Sejroe, but the overall within-herd prevalence was low (11.5% and 7.7% for cut-off titres of 1:30 and 1:100, respectively). The in-herd prevalence of leptospirosis in the sow unit (determined for 113/140 animals) was high when using a lowered cut-off threshold (32.7% vs. 1.8% for cut-off titre of 1:30 and 1:100, respectively). In this unit, the most prevalent serogroup was Autumnalis. The final diagnostic confirmation of Leptospira sp. maintenance within the farm was obtained through detection by PCR of Leptospira sp. DNA in an aborted swine litter. Despite the fact that a common causative infective agent was diagnosed in both species, the direct link between the two animal units was not found. Factors such as drinking from the same water source and the use of manure prepared with the swine slurry might raise suspicion of a possible cross-contamination between the two units. In conclusion, this work suggests that leptospirosis be included in the differential diagnosis of reproductive disorders and spontaneous abortions in production animals and provides data that justify the use of a lowered threshold cut-off for herd diagnosis.

Serosurvey of anti- Leptospira sp. and anti- Toxoplasma gondii antibodies in capybaras and collared and white-lipped peccaries.

Brazilian native species are reemerging as increasingly free-ranging populations. Sera from 31 capybaras (Hydrochoerus hydrochaeris) and 28 peccaries (Pecari tajacu and Tayassu pecari) were tested for anti-Leptospira and anti-Toxoplasma gondii antibodies using microscopic seroagglutination test. Nineteen percent of free-ranging and 10.0% of captive capybaras, along with 31.8% of collared peccaries, were seropositive for T. gondii. None was seropositive for Leptospira sp. The present findings indicated low risk of disease, particularly among capybaras and white-lipped peccaries; however, active surveillance programs are important for monitoring wildlife health and public health once they are in public parks around cities.

Polymorphisms of TLR4 Asp299Gly and TNF-α -308G/A in Leptospirosis

AbstractBackground : TLR4 Asp299Gly and TNF-α -308G/A polymorphisms have been shown to be associated with increased susceptibility and severity of infection. TLR4 Asp299Gly polymorphism could affect the host’s ability to respond to leptospira sp. TNF-α -308G/A polymorphism, is associated with the high producer of TNF-α.Methods : Total of 36 leptospirosis patients (IgM anti leptospira and MAT positive) and healthy individual with the equal number were included. The polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using site spesific restriction enzyme.Results : Distribution of homozygous wild-type TLR4 Asp299Gly polymorphism was higher in both of groups ( 94.5:97.2%.) and homozygous mutant allele was absent. There was not significantly difference of TLR4 Asp299Gly in leptospirosis patients and healthy group ( ρ=1.00; OR 0.5; 95%CI, 0.04-5.6) and between mild and severe leptospirosis (ρ=0.54; OR 1.54 ; 95% CI, 1.20-1.98). The presence of homozygous wild-type TNF-α -308G/A polymorphism was higher between leptospirosis patients and healthy group (100:94.5%) andhomozygous mutant allele was not found in both of the groups. No significantly different of TNF-α -308G&gt;A polymorphism between leptospirosis patient and healthy group (ρ=0.49).Conclusions : In this study, the polymorphisms of TLR4 Asp299Gly and TNF-α -308G/A were not associated with the susceptibility and severity of leptospirosis. Keywords : Leptospirosis, TLR4 Asp299Gly polymorphism, TNF-α -308G/A polymorphism

P05.03 Performance of two enzyme and one strip immune assay for the detection of igm anti-treponema pallidumantibodies

Background The detection of anti- Treponema pallidum (Tp) IgM may be useful in the diagnosis of very early syphilis, re-infection and in the assessment of the newborn. We aimed to evaluate two enzyme and one strip immune assay for the detection of anti-Tp IgM in blood specimens. Methods A total of 33 blood specimens collected from 16 patients presenting with a syphilitic ulcer were analysed. The specimens were tested employing: Rapid Plasma Reagin; Treponema Pallidum Particle Agglutination (TPPA); recomLine Treponema IgM (Mikrogen Diagnostik); recomWell Treponema IgM (Mikrogen Diagnostik); Anti- Treponema pallidum ELISA -IgM (Euroimmun). Ulcer specimens were tested by qPCR. A specimen was considered positive for anti-Tp IgM when both TPPA and IgM, obtained with any of the assays, were positive. Specimens containing antibodies against Epstein Barr Virus, Leptospira sp, Borrelia sp, Plasmodium sp, Herpes Simplex Virus 1 and 2 and specimens from pregnant women were tested with the three IgM assays to determine cross reactivity. Results Anti-Tp IgM was found in 22/33 specimens. The recomLine assay detected 17/22 (77%), the recomWell 15/22 (68%) and the Euroimmun 14/22 (64%) positive specimens. None of the three assays provided false positive results. Borderline results were obtained with the recomLine (N = 6) and the recomWell (N = 2) assay. Considering the borderline results as positive increased the detection rate to 95% for the recomLine and to 72% for the recomWell. However, two and one false positive result was then obtained with the recomLine and recomWell assay, respectively. Tp DNA was detected in 13/16 (81%) ulcer specimens. The Euroimmun assay cross reacted with malaria antibodies in one sample. Conclusions None of the three assays showed to be highly sensitive. Surprisingly, the highest sensitivity was obtained with the recomLine assay. The sensitivity improved by defining the borderline results as positive but decreased the specificity. Disclosure of interest The IgM assays were kindly provided by the two companies, Euroimmun and Mikrogen Diagnostik. No other funding was received for this work.

Presence of leptospires on genital tract of mares with reproductive problems

Leptospirosis is a zoonotic disease of global importance, and has a worldwide distribution. Equine leptospirosis is commonly manifested by recurrent uveitis, reproductive disorders, as abortions, embryonic absorption, stillbirth and the birth of weak foals. The aim of this study was to verify the presence of Leptospira sp or its DNA in genital tract of mares with reproductive problems. A total of 38 mares with reproductive problems were studied. All the mares were sampled for blood (for serology), urine (for culturing and qPCR), vaginal fluid-VF and endometrial biopsy-EB (for culturing, qPCR and indirect immunofluorescence). PCRs products were sequenced for secY gene. Seventeen (44.7%) serum samples were reactive, predominantly against serogroups Australis (76.4%) and Pomona (23.6%). No positive culture was obtained, but DNA was detected by qPCR on urine samples (26.3%), VF (44.7%) and EB (18.4%) collected 2 months or longer following diagnosis of early fetal death and endometritis. Leptospira cell aggregations were visible by indirect immunofluorescence on 57.1% (4/7) EBs and 17.6% (3/17) VFs. A total of 18 amplicons showed interpretable sequences. Out of those 18 amplicons, 15 presented 100% of identity with the species L. interrogans (sv Bratislava and Pomona), while three were L. borgpertersenii. This study suggests the presence of leptospires in the uterus of mares with reproductive problems. Moreover, serology was shown not to be indicated for the diagnosis of presumptive Leptospira infection in early gestation. The most common agent of the genital infection in those mares was L. interrogans, most probably sg Australis.

Presence of antibodies against Leptospira serovars in Chaetophractus villosus (Mammalia, Dasypodidae), La Pampa province, Argentina

Leptospirosis is a zoonosis of worldwide distribution. The aim of this study was to examine the presence of antibodies against 21 Leptospira reactive serovars in Chaetophractus villosus in La Pampa province, Argentina, using the microscopic agglutination test (MAT). Pathologic changes compatible with leptospirosis and in situ detection of the agent by immunohistochemistry were studied in 24 and 3 individuals respectively. Only 35/150 (23.3%) serum samples had antibodies against Leptospira sp. Six percent of the samples reacted with serovar Canicola, 4.7% with serovar Castellonis, 1.3% with serovar Icterohemorrhagieae and 0.7% with serovar Hardjo. Sixteen (10.6%) serum samples agglutinated with Castellonis–Icterohemorrhagiae and Canicola–Castellonis serovars, both with 4.7%, and Canicola–Hardjo and Castellonis–Canicola–Icterohemorrhagiae both with 0.6%. Fourteen animals had variable degrees of lesions, which were more severe in animals with higher serological titers (3200), and Leptospira sp. was detected in 3 animals by immunohistochemistry. These results represent the first record of the presence of Leptospira in C. villosus in La Pampa.

Microscopic changes in the kidneys of cows infected with Leptospira sp.

Abstract The article presents data on histopathological studies of the kidneys of cows, which either recovered or died from leptospirosis. Fragments of seven kidneys from slaughtered cows, positive for Leptospira antibodies in the microscopic agglutination test (MAT) (titres of 50 and higher) were used in the study. The MAT was conducted with eight serological groups of Leptospira: Canicola, Grippotyphosa, Hebdomadis, Icterohaemorrhagiae, Pomona, Sejroe, Tarassovi, and Australis. Microscopic changes in all morphological structures of the kidneys were presented. Micromorphological criteria, which can be used for post-mortem diagnosis of leptospirosis were established. They included: serous glomerulonephritis with granular dystrophy of podocytes, necrosis and collapse of the inner layer of Bowman's capsule, partial destruction of capsule and vascular glomeruli, granular and vacuolar degeneration and destruction of tubular epithelial cells, foci of interstitial oedema, and infiltrations predominantly with monocytes and isolated neutrophils. Microscopic changes in the kidneys suggest that the cows died from leptospirosis as a result of toxic shock syndrome.