Ocorrência de anticorpos contra Leptospira spp. em eqüinos de área urbana de Londrina, Paraná, Brasil

A total of 320 horses were studied in this paper, both male and female, between two and 17 years of age, which were used for traction of wagons in the urban area of the municipality of Londrina (PR). These animals were kept, after their daily work, in abandoned areas or plots, in the outskirts of the urban area of the city. When these animals were attended by the veterinarians, between 1996 and 2005, none of them presented symptoms suggesting leptospirosis. The most frequent reasons for the visit were loss of weight, unwillingness for work, parasitism, laminess, and wounds. Microscopic Seroagglutination Test (SAM), with 22 Leptospira serovars, was performed in sera sample from all these animals. The aim of this study was to investigate the occurrence of antibodies against Leptospira spp. in horses from the urban area of Londrina (PR). From the samples tested, 214 (66.88%) were considered positive, with titers between 100 and 3200, being that 49 (22.90%) presented antibodies against a single serovar of Leptospira, and 165 (77.10%) samples presented antibodies against two or more serovars simultaneously, where in 88 (53.33%) it was possible to characterize the most likely probable serovar. Antibodies against the serovar Icterohaemorrhagiae were detected in 32 (23.36%) animals.Foram estudados 320 eqüinos, machos e fêmeas com idade entre dois a 17 anos, utilizados para tração de carroças na área urbana do município de Londrina (PR). Estes animais eram mantidos, após a jornada diária de serviço, em áreas ou terrenos baldios, localizados na periferia da área urbana do município. Por ocasião do atendimento, realizado entre 1996 e 2005, no Hospital Veterinário da Universidade Estadual de Londrina, nenhum dos animais apresentou sintomas sugestivos de leptospirose, sendo os motivos mais freqüentes, o emagrecimento, a indisposição para o trabalho, a verminose, problemas no sistema locomotor e feridas. A prova de soroaglutinação microscópica (SAM), com 22 sorovares de Leptospira, foi realizada em amostra de soro de todos os animais. O objetivo deste trabalho foi investigar a ocorrência de anticorpos contra Leptospira spp. em eqüinos da área urbana do município de Londrina. Foram consideradas positivas 214 (66,88%) amostras de soros com títulos entre 100 e 3200, sendo em 49 (22,90%) detectadas anticorpos contra um único sorovar de Leptospira e em 165 (77,10%) amostras com anticorpos contra dois ou mais sorovares simultâneamente, sendo em 88 (53,33%) destes caracterizado o sorovar mais provável. Anticorpos contra o sorovar Icterohaemorrhagiae foram detectados em 32 (23,36%) animais

Acompanhamento sorológico e molecular do sorovar hardjo no diagnóstico de leptospirose em um rebanho bovino leiteiro

The cattle are considered hosts of the Hardjo serovar, causing economic damages due to the reproductive failures like abortions and infertility. The serovar Hardjo usually remains in the reproductive tract and also in the renal tubules where it is eliminated intermittently in the urine for months. Placental remnants, the aborted fetus and contaminated urine promote the permanence of this bacterium within the herd for years. Thus, the objective of this study was to monitor for prolonged period, cows naturally infected with Leptospira ssp. through microbiological culture, serological examination and DNA detection of the pathogen in the urine. The dairy herd was composed of 50 breeding cows with a history of abortion and infertility, without leptospirosis vaccine and located in the northern region of Paraná. Blood and urine samples were collected and laboratorial diagnosis were performed five times at intervals of four months. Blood samples were collected from the all 50 animals and the serum was submitted to the microscopic agglutination test (MAT) for the detection of anti-leptospira antibodies. Of the total cows, 20 showed antibody titres ? 1: 100 in MAT and urine samples were collected from only those animals with higher titers to perform nested-PCR (n-PCR) and bacterial isolation per culture. In addition, two urine samples from five animals with antibody titers < 1: 100 were collected in MAT for n-PCR. Serovar Hardjo was considered the most frequent during the serological monitoring of the animals evaluated. The n-PCR technique was able to detect leptospiral DNA in the urine of animals with MAT ? 1: 100 antibody titers and urine from animals whose titers were < 1: 100. Sequencing of the leptospiral amplicons shared 100% nucleotide sequence identity with the Leptospira interrogans species. Positive n-PCR results from animals with titers of < 1: 100 suggest that the cut-off of MAT is could be not sufficient to detect renal carriers, so it is also important to use n-PCR as an additional diagnostic tool for identify infected animals with Hardjo serovar and whose serology was negative.Os bovinos são considerados hospedeiros do sorovar Hardjo, causando prejuízos econômicos devido aos problemas reprodutivos como abortos e infertilidade. O sorovar Hardjo costuma permanecer no trato reprodutivo e também nos túbulos renais onde é eliminado de forma intermitente na urina por meses. Restos placentários, o feto abortado e a urina contaminada favorecem a permanência dessa bactéria dentro do rebanho por anos. Assim, o objetivo deste estudo foi monitorar por período prolongado, vacas naturalmente infectadas por Leptospira ssp. através de cultura microbiológica, exame sorológico e detecção de DNA do patógeno na urina. O rebanho leiteiro estudado era constituído por 50 vacas reprodutoras com histórico de abortos e infertilidade, sem uso de vacina contra a leptospirose e localizada na região Norte do Paraná. As amostras de sangue e urina foram coletadas e submetidas a análises laboratoriais em cinco vezes em intervalos espaçados de quatro meses. As amostras de sangue foram coletadas dos 50 animais e o soro foi submetido ao teste de soroaglutinação microscópica (SAM) para detecção de anticorpos anti-leptospira. Do total vacas, 20 demonstraram títulos de anticorpos ? 1: 100 na SAM e foram coletadas amostras de urina apenas destes animais com maiores título para realizar a nested-PCR (n-PCR) e isolamento bacteriano por cultura. Adicionalmente, foram coletadas duas amostras de urina de cinco animais com títulos de anticorpos < 1: 100 na SAM para n-PCR. O Sorovar Hardjo foi considerado como sendo o mais frequente durante o monitoramento sorológico dos animais avaliados. A técnica de n-PCR foi capaz de detectar DNA leptospiral na urina de animais com títulos de anticorpos ? 1: 100 e na urina de animais cujos títulos eram < 1:100. O sequenciamento dos amplicons leptospíricos compartilhou 100% de identidade da sequência de nucleotídeos com a espécie Leptospira interrogans. Os resultados positivos da n-PCR dos animais com títulos < 1: 100 sugerem que a diminuição do ponto de corte da SAM pode não ser suficiente para detectar os portadores renais, por isso também é importante usar a n-PCR como uma ferramenta diagnóstica adicional para identificar os animais infectados com serovar Hardjo e cuja sorologia foi negativa

Epidemiological status of bovine leptospirosis in the State of Paraná, Brazil

The objective of this study was to determine the prevalence of antibodies against Leptospira spp and their geographic distribution and to identify the risk factors associated with this disease in cattle herds with reproductive activity in the state of Paraná. A total of 14 163 females aged ?24 months originating from 1 926 herds that were not vaccinated against Leptospira spp were evaluated. To detect the Leptospira spp antibodies, all serum samples were submitted for a microscopic serum-agglutination test (MAT). An epidemiological questionnaire was performed at each farm visited to characterize the management practices and study the risk factors associated with the presence or absence of Leptospira spp. To perform geoprocessing, the geographical coordinates of each farm were recorded with a Global Positioning System (GPS), which were then used to create a map. The apparent prevalence of bovine leptospirosis in the herds was the parameter used to generate the final risk map. The apparent prevalences of seropositivity in the herds and animals in Paraná state were 54.28%(95%CI:52.05- 56.51) and 37.70%(95%CI:34.54-40.86), respectively. Multiple logistic regression analysis revealed that the risk factors associated with leptospirosis due to any serovar were the rental of grazing areas [OR=1.91(1.36-2.68)], presence of horses [OR=1.59(1.28-1.97)], presence of > 20 females ? 24 months of age [OR=2.25(1.46-3.49)], presence of > 49 bovines [OR=2.78(1.82-4.26)], purchase of animals for reproduction [OR=1.96(1.59-2.41)] and presence of calving pens [OR=1.32(1.04-1.68)]. Risk factors for leptospirosis due specifically to serovar Hardjo were the presence of > 49 bovines [OR=3.05(1.94-4.80)], presence of > 20 females ? 24 months [OR=2.38(1.50-3.79)], presence of horses [OR=1.87(1.45-2.43)], purchase of animals for reproduction [OR=2.14(1.68-2.72)] and rental of grazing areas [OR=2.22(1.54-3.21)]. Geographically, seropositivity to Leptospira spp by MAT identified the regions North/Northwest and Southwest in the state as the areas with a higher risk of disease occurrence

Leptospires detection in kidney, liver and uterus of cows slaughtered in Paraná State, Brazil Detecção de leptospiras em rim, fígado e útero de fêmeas bovinas abatidas no estado do Paraná, Brasil

The objective of this trial was to detect leptospires in fragments of kidney, liver and uterus of 96 cows with unknown sanitary status, randomly chosen at slaughter in Paraná, Brazil. All 96 urine samples were submitted to direct examination using dark field microscopy. Positive samples in the direct examination and all kidney, liver and uterus fragments were cultured in EMJH modified medium and Tween 80/40/LH. Sections cut of kidney, liver and uterus were stained by Hematoxylin-Eosin (HE) and indirect immunoperoxidase with hyperimmune serum against serovar Hardjo (Hardjoprajitno) and strain Londrina 14 (LO14), from serovar Canicola, which was isolated in the northern region of Paraná state, Brazil, used as primary antibodies. Direct examination detected leptospires in the urine of four animals. All attempts to isolate leptospires from urine and kidney, liver and uterus fragments were negative after 16 weeks of incubation. In the HE stain, focal infiltrate of mononuclear inflammatory cells was observed in the renal interstitial area of most animals. In the indirect immunoperoxidase assay using hyperimmune serum against LO14 strain, the kidney of only one animal presented positive results. All fragments of kidney, liver and uterus tested with hyperimmune serum against serovar Hardjo were negative.<br>O objetivo deste trabalho foi detectar leptospiras em fragmentos de rim, fígado e útero de 96 fêmeas bovinas com histórico sanitário desconhecido, escolhidas aleatoriamente durante o abate em um frigorífico no Estado do Paraná, Brasil. Todas as 96 amostras de urina foram submetidas ao exame direto em microscópio de campo escuro. As amostras positivas neste exame e todos os fragmentos de rim, fígado e útero foram semeados nos meios de cultura EMJH modificado e Tween 80/40/LH. Os cortes histológicos de rim, fígado e útero foram submetidos à coloração de Hematoxilina-Eosina (HE) e a prova de imunoperoxidase indireta com soros hiperimunes contra o sorovar Hardjo (Hardjoprajitno) e contra a estirpe Londrina (LO14), do sorovar Canicola, isolada no norte do Paraná, Brasil, utilizados como anticorpo primário. O exame direto detectou leptospiras na urina de quatro animais. Todas as tentativas de isolamento das urinas e fragmentos de rim, fígado e útero foram negativas após 16 semanas de incubação. No exame histopatológico foi observado infiltrado focal de células mononucleares no interstício renal da maioria dos animais. Na prova de imunoperoxidase indireta com o soro hiperimune contra a estirpe LO14, o rim de um animal foi o único a apresentar resultado positivo. Todos os fragmentos de rim, fígado e útero testados com o soro hiperimune contra o sorovar Hardjo foram negativos

Association between microscopic kidney´s lesions and presence of antibodies against Leptospira spp in apparently healthy swine, originated from slaughterhouse in northern Paraná state

O objetivo deste trabalho foi demonstrar a associação entre lesões renais microscópicas e a presença de anticorpos contra Leptospira spp em suínos aparentemente sadios, abatidos em frigorífico da região norte do Estado do Paraná. Foram colhidas amostras de rim e coágulo sanguíneo de 120 animais com lesões renais macroscópicas tipo white spots e de 120 sem lesões renais macroscópicas tipo White spots, utilizados como controle. As amostras de soro foram submetidas à prova de soroaglutinação microscópica (SAM) e as amostras de rim, ao exame histopatológico corado pela Hematoxilina-Eosina (HE). Embora nenhuma associação (p=0,27) entre as lesões renais macroscópicas tipo white spots e a presença de anticorpos contra Leptospira spp tenha sido observada, verificou-se uma significativa associação (p=0,015) entre a nefrite intersticial e a soropositividade para leptospirose.The aim of this work was to demonstrate the association between microscopic kidney´s lesions and presence of antibodies against Leptospira spp in apparently healthy swine from slaughterhouses located in northern Paraná State. Samples of kidney and blood coagulum were collected from 120 animals with macroscopic kidney´s lesions type white spots and from120 animals without macroscopic lesions, used as control. Samples of serum, were submitted to microscopic agglutination test (MAT) and samples of kidney for histopathological evaluation stained with Hematoxylin and Eosin (HE). Although no association (p=0,27) between macroscopic kidney’s lesions type white spots and presence of antibodies against Leptospira spp has been observed, there was a significant association (p=0,015) between interstitial nephritis and seropositivity to leptospirosis

Empagliflozin in Patients with Chronic Kidney Disease

Background The effects of empagliflozin in patients with chronic kidney disease who are at risk for disease progression are not well understood. The EMPA-KIDNEY trial was designed to assess the effects of treatment with empagliflozin in a broad range of such patients. Methods We enrolled patients with chronic kidney disease who had an estimated glomerular filtration rate (eGFR) of at least 20 but less than 45 ml per minute per 1.73 m(2) of body-surface area, or who had an eGFR of at least 45 but less than 90 ml per minute per 1.73 m(2) with a urinary albumin-to-creatinine ratio (with albumin measured in milligrams and creatinine measured in grams) of at least 200. Patients were randomly assigned to receive empagliflozin (10 mg once daily) or matching placebo. The primary outcome was a composite of progression of kidney disease (defined as end-stage kidney disease, a sustained decrease in eGFR to &lt; 10 ml per minute per 1.73 m(2), a sustained decrease in eGFR of &amp; GE;40% from baseline, or death from renal causes) or death from cardiovascular causes. Results A total of 6609 patients underwent randomization. During a median of 2.0 years of follow-up, progression of kidney disease or death from cardiovascular causes occurred in 432 of 3304 patients (13.1%) in the empagliflozin group and in 558 of 3305 patients (16.9%) in the placebo group (hazard ratio, 0.72; 95% confidence interval [CI], 0.64 to 0.82; P &lt; 0.001). Results were consistent among patients with or without diabetes and across subgroups defined according to eGFR ranges. The rate of hospitalization from any cause was lower in the empagliflozin group than in the placebo group (hazard ratio, 0.86; 95% CI, 0.78 to 0.95; P=0.003), but there were no significant between-group differences with respect to the composite outcome of hospitalization for heart failure or death from cardiovascular causes (which occurred in 4.0% in the empagliflozin group and 4.6% in the placebo group) or death from any cause (in 4.5% and 5.1%, respectively). The rates of serious adverse events were similar in the two groups. Conclusions Among a wide range of patients with chronic kidney disease who were at risk for disease progression, empagliflozin therapy led to a lower risk of progression of kidney disease or death from cardiovascular causes than placebo