Prevalence and risk factors for Leptospira spp. in cattle herds in the south central region of Paraná state

The aim of this study was to determine the prevalence of anti-Leptospira spp. antibodies and the risk factors for Leptospira spp. infection in breeding cattle herds in the south central region of Paraná state. It was based on the statistic delineation/serological samples and information regarding the selected farms employed in the study of bovine brucellosis for Paraná state in the context of National Program for Control and Eradication of Brucellosis and Tuberculosis. A total of 1.880 females aged >24 months from 274 non vaccinated herds were studied. Serum samples were tested for antibodies against Leptospira spp. using microscopic agglutination test (MAT) with 22 Leptospira serovars. The epidemiological questionnaire was applied on all the selected farms and aimed to obtain epidemiological data. Hundred eighty one of 274 herds were positive for Leptospira spp./presenting prevalence of positive herds of 66.06% (IC95%=60.12-71,65%). Presence of >43 cattle (OR=3.120; IC=1.418-6.867)/animal purchase (OR=2.010; IC=1.154-3.500)/rent of pastures (OR=2.925; IC=1.060-8.068) and presence of maternity paddock (OR=1.981; IC=1,068-3,676) were identified as risk factors for leptospirosis due to any serovar in the multivariate logistic regression. Risk factors for leptospirosis due to serovar Hardjo were presence of >43 cattle (OR=3.622; IC=1.512-8,677)/animal purchase (OR=3.143; IC=1.557-6.342)/rent of pastures (OR=4.070; IC=1.370-12.087) and presence of horses (OR=2.981; IC=1.321-6.726). These results indicate that Leptospira spp. infection is widespread in the south central region of Paraná state and that factors related to the herd characteristic and management are associated with the infection.

Frequency of Leptospira spp. in sheep from Brazilian slaughterhouses and its association with epidemiological variables

Leptospirosis is a worldwide anthropozoonosis that infects livestock, including sheep as the carriers to other animals and humans. The present study aimed to determine the prevalence of Leptospira spp. in sheep from two slaughterhouses in the state of São Paulo, Brazil and its association with epidemiological variables. Serum samples from 182 sheep were evaluated for Leptospira spp. antibodies by microscopic agglutination test (MAT). Results indicated 34/182 (18.68%; CI95% 13.70-24.98%) positive serum samples, mainly to the serovar Copenhageni (17/34; 50%; CI95% 33.99-66.01%). Bacterial growth in the Fletcher medium was detected for 13/34 (38.24%; CI95% 23.87-55.08%) animals, and confirmed by Polymerase Chain Reaction (PCR) and sequencing for only two kidney samples from two animals. Thus, treatment and vaccination of sheep, besides rodent control, can be useful to prevent the infection in the studied region since sheep are important Leptospira spp. carriers, and its transmission to slaughterhouse workers is mainly through the manipulation of visceral tissues.

Controle da leptospirose em bovinos de leite com vacina autógena em Santo Antônio do Monte, Minas Gerais

Um surto de leptospirose foi observado em bovinos leiteiros em Santo Antônio do Monte, Minas Gerais. O rebanho apresentava reações positivas anti-leptospira sorovar Hardjo no teste de microaglutinação (MAT) e havia sido vacinado anteriormente com vacina experimental contendo a sorovariedade Hardjo. O MAT revelou 48,06% dos bovinos positivos para sorovariedade Hardjo genótipo Hardjobovis, 36,82% para sorovariedade Hardjo genótipo Hardjoprajitno. Os animais apresentavam aborto e mastite com presença de sangue no leite. A presente pesquisa teve como objetivos isolar as sorovariedades existentes a partir da urina de vacas sorologicamente positivas, elaborar uma vacina experimental com as sorovariedades isoladas no rebanho, avaliar a eficiência do programa de vacinação por um período de dois anos por meio da sorologia do rebanho. Foi isolada Leptospira spp. a partir da urina de duas vacas com sinais sugestivos da doença. As amostras isoladas foram identificadas pela sorologia com anticorpos monoclonais e seqüenciamento do gene 16S rRNA como pertencentes à espécie Leptospira interrogans, sorogrupo Sejroe, sorovariedade Hardjo e genótipo Hardjoprajitno. O uso da vacina autógena foi eficaz no controle da leptospirose no rebanho no período de dois anos. Os resultados da sorologia revelaram ausência de animais positivos na última prova realizada no rebanho.

Serologic and molecular diagnostic and bioassay in mice for detection of Toxoplasma gondii in free ranges chickens from Pantanal of Mato Grosso do Sul

The aim of this study was to investigate the occurrence of Toxoplasma gondii and compare the results obtained in the Modified Agglutination Test (MAT), Polimerase Chain Reaction (PCR) and bioassay in mice. In order to accomplish this, 40 free-range chickens from eight farms in neighboring areas to the Pantanal in Nhecolândia, Mato Grosso do Sul, were euthanized and blood samples, brain and heart were collected. The occurrence of anti-T. gondii antibodies found in chickens was 67.5% (27 samples), considering as a cutoff point the dilution 1:5. Among the samples analyzed, 7 (25.9%) were positive in the dilution 1:5, 3 (11.1%) in 1:10, 2 (7.4%) in 1:20, 3 (11.1%) in 1:320, 1 ( 3.7%) in 1:640, 3 (11.1%) in 1:1280, 2 (7.4%) in 1:2560, 4 (14.8%) in 1:5120 and 2 (7.4%) in 1:10.240. From the mixture of tissue samples (brain and heart) from the chickens analyzed, 16 (40%) presented electrophoretic bands compatible with T. gondii by PCR (gene B1). In the comparison of techniques, 59.26% positivity in PCR was revealed among animals that were seropositive in MAT (cutoff 1:5). From 141 inoculated mice, six (4.44%) died of acute toxoplasmosis between 15 and 23 days after inoculation. Surviving mice were sacrificed at 74 days after inoculation, and a total of 28 cysts were found in the brains of 10 distinct groups. From the seropositive hens, 27 bioassays were performed and 11 (40.7%) isolates were obtained. A greater number of isolations happened in mice that were inoculated with tissues from chickens that had high titers for anti-T. gondii antibodies. Chronic infection in mice was observed in nine groups (33.3%) from five different properties. Among the surviving mice, 25.6% were positive for T. gondii in MAT (1:25). From mice positive in PCR, 87.5% were also positive in MAT. Among the PCR-negative mice, 5.2% were positive for T. gondii in MAT. It can be concluded through this study that the occurrence of infecton by T. gondii in the rural properties studied was high, that PCR directed to gene B1 does not confirm the viability of the parasite, but it can be used as a screening method for the selection of chickens infected by T. gondii, that the animals with titer greater than 10 must be prioritized for the selection of animals for bioassay, since for them, the chances of isolating the parasite are greater and that seroconversion in experimentally infected mice is not a good indicator for isolating the agent.

Epidemiology of leptospirosis at Sorocaba Zoo, São Paulo state, Southeastern Brazil

Leptospirosis is considered a worldwide distributed zoonosis, caused by the bacteria Leptospira spp. Since several species of wildlife animals are reportedly reservoirs, the aim of the present study was to know the epidemiology of leptospirosis at the Sorocaba Zoo, Southern Brazil. Serum samples of wild mammals from Artiodactyla, Carnivora, Didelphimorphia, Diprotodontia, Perissodactyla, Pilosa, Primates, Proboscidea and Rodentia orders, kept in captivity as well as from zoological staff were assayed by microscopic agglutination test (MAT). Whole blood, urine and tissue samples from wild mammals and synanthropic animals were assayed by polymerase chain reaction (PCR). An epidemiological survey was applied to evaluate the risk factors for animal infection and staff level of knowledge on leptospirosis. A total of 13/229 (5.68%; CI95% 3.37-9.47%) serum samples from wild mammals were reagent on MAT. Serology from synanthropic animals, zoo staff and molecular analysis of animal samples were all negative. Leptospirosis knowledge of zoo park staff was considered medium. In conclusion, leptospiral infection occurs at the studied zoo but due to the low occurrence found, the lowest reported in literature, wild captive mammals do not act as source of infection of leptospirosis to other animals and human beings.

Frequency of Toxoplasma gondii antibodies in tufted capuchin monkeys (Cebus apella nigritus) from an ecological station in the State of São Paulo, Brazil

Toxoplasmosis is a worldwide zoonosis caused by Toxoplasma gondii, an obligate intracellular parasite protozoan. A large percentage of animals presents specific antibodies caused by a previous exposition, resulting in a chronic infection. Felides are the definitive hosts and the other warm-blooded animals, including primates, are the intermediate hosts. This study was aimed to determine the prevalence of T. gondii infection in free-living tufted capuchin monkeys (Cebus apella nigritus) from an ecological station located on Mata de Santa Teresa, Ribeirão Preto, SP, Brazil. T. gondii antibodies were analyzed by modified agglutination test (MAT) in serum samples of 36 tufted capuchin monkeys, considering eight as cut-off titer. From the studied animals, 3/36 (8.33%; CI95% 3.0-21.9%) presented T. gondii antibodies, all with titer 32. No significative difference was observed relating to the sex (1/3 male and 2/3 female), and to the age (1/3 young and 2/3 adult) (P>0.05). Thus, these results demonstrate the presence of T. gondii antibodies in primates from São Paulo state.

The importance of Leptospira interrogans serovars Icterohaemorrhagiae and Canicola in coastal zone and in southern fields of Rio Grande do Sul, Brazil

This study aimed to describe the occurrence of Leptospira interrogans serovars Icterohaemorrhagiae and Canicola, in coastal zone and in southern grasslands of Rio Grande do Sul, Brazil. In each one of the four analyzed farms blood samples were collected from free-living wild animals, domestic animals and humans to perform serological testing for leptospirosis. The presence of antibodies was verified by microscopic agglutination test (MAT). The criterion adopted to consider a serum as agglutination reactant was at least 50% of leptospira for a microscopic field of 100x. From 17 blood samples collected at Chuí, five (29.41%) were positive, three (60.00%) for serovar Icterohaemorrhagiae and two (40.00%) for Canicola. From 21 samples collected in the County of Santana da Boa Vista, six (28.57%) were positive, four (66.67%) for serovar Canicola and two (33.33%) for serovar Icterohaemorrhagiae. From 32 samples collected at Alegrete, 10 (31.25%) were positive, seven (70.00%) for serovar Icterohaemorrhagiae and three (30.00%) foro serovar Canicola. From 17 blood samples collected in Cruz Alta, three (17.64%) were positive, two (66.67%) for serovar Icterohaemorrhagiae and one (33.33%) for Canicola. It is necessary to improve sanitary practices on farms in the state of Rio Grande do Sul, in order to achieve success in leptospirosis control programs.

The role of alveolar type II cells in swine leptospirosis

Abstract: This study aimed to investigate a possible relationship between alveolar type II cells and the inflammatory response to infection with Leptospira spp., and thus comprise a further element that can be involved in the pathogenesis of lung injury in naturally infected pigs. The study group consisted of 73 adult pigs that were extensively reared and slaughtered in Teresina, Piauí state, and Timon, Maranhão state, Brazil. The diagnosis of leptospirosis was made using the microscopic agglutination test (MAT) aided by immunohistochemistry and polymerase chain reaction. The MAT registered the occurrence of anti-Leptospira antibodies in 10.96% (8/73) of the pigs. Immunohistochemistry allowed for the visualization of the Leptospira spp. antigen in the lungs of 87.67% (64/73) of the pigs. There was hyperplasia of bronchus-associated lymphoid tissue and circulatory changes, such as congestion of alveolar septa, parenchymal hemorrhage and edema within the alveoli. Lung inflammation was more intense (p = 0.0312) in infected animals, which also showed increased thickening of the alveolar septa (p = 0.0006). Evaluation of alveolar type II (ATII) cells using an anti-TTF-1 (Thyroid Transcription Factor-1) antibody showed that there were more immunostained cells in the non-infected pigs (53.8%) than in the infected animals (46.2%) and that there was an inverse correlation between TTF-1 positive cells and the inflammatory infiltrate. There was no amplification of Leptospira DNA in the lung samples, but leptospiral DNA amplification was observed in the kidneys. The results of this study showed that a relationship exists between a decrease in alveolar type II cells and a leptospire infection. Thus, this work points to the importance of studying the ATII cells as a potential marker of the level of lung innate immune response during leptospirosis in pigs.

Occurrence of antibodies against Leptospira spp. In free-ranging wild canids from the Brazilian savanna

Abstract: The Brazilian savanna, also known as Cerrado, is one of the world's richest and most ecologically invaluable tropical savanna regions. There are few studies in Brazil about the diseases that affect the wild canids of this biome, which may be harmful to wildlife populations and public health. The aim of this study was to evaluate the occurrence of antibodies against Leptospira spp. in three Cerrado wild canids species using the microscopic agglutination test (MAT). Serum samples were tested from 19 crab-eating foxes (Cerdocyon thous), 14 maned wolves (Chrysocyon brachyurus), and seven hoary foxes (Lycalopex vetulus), all free-ranging animals found in the municipalities of Araguari and Uberlândia, Minas Gerais State, and Cumari, Goiás State, Brazil. Fourteen (35%) of these samples were seropositive. The most frequent serovars detected in the samples were Copenhageni and Hardjo, but reactions to the serovars Autumnalis, Grippotyphosa, Hebdomadis, Wolffi, and Icterohaemorrhagiae also occurred. Notwithstanding other reported results, this study is the first to report the presence of antibodies against Leptospira spp. in L. vetulus. The three species of wild canids examined may act as potential hosts for several serovars of leptospira in Brazil's savanna environment.

Ocorrência de anticorpos anti-Toxoplasma gondii em aves silvestres de três Unidades de Conservação Federais da Paraíba e da Bahia

Resumo: A vigilância e monitoramento de doenças em animais silvestres são imprescindíveis no contexto ambiental e de saúde pública, pois estes animais agem como sentinelas, refletindo alterações ambientais precocemente, o que proporciona maior eficácia no monitoramento ambiental e permite o acesso rápido a informações sobre as condições da área. Neste contexto, as aves são importantes no ciclo biológico do Toxoplasma gondii e na epidemiologia da toxoplasmose, principalmente porque seus tecidos representam importantes fontes de proteína na alimentação de felídeos e humanos. Objetivou-se detectar anticorpos anti-T. gondii, por meio do teste de aglutinação modificada em aves silvestres de três Unidades de Conservação (UC) Federais dos Estados da Paraíba e Bahia. No período de dezembro de 2011 a outubro de 2013 foram capturadas com redes de neblina 222 aves silvestres pertencentes a 67 espécies, 27 famílias e 12 ordens. Após a captura, foi colhido sangue de cada animal e separado o soro, que foi submetido ao Teste de Aglutinação Modificada (MAT≥1:25) utilizando taquizoítos inativados na formalina e 2-mercaptoetanol. Dentre as 222 amostras analisadas, três (1,3%) foram sororreagentes: 1 de 16 (6,2%) pipira-preta Tachyphonus rufus (título 50), 1 de 5 (20%) juriti-gemedeira Leptotila rufaxilla (título 50) e 1 de 1 (100%) caneleiro-enxofre Casiornis fuscus (título 25). Este é o primeiro relato da ocorrência de anticorpos anti-T. gondii nas referidas espécies de aves silvestres de vida livre nas duas UC Federais estudadas.

Seletividade de herbicidas para híbridos simples de milho (Zea mays L.)

Com o objetivo de estudar a seletividade de herbicidas, aplicados isolados ou em mistura, foram realizados quatro experimentos em vasos, com híbridos simples de milho IAC 7777 (dentado), C-72 (dentado) e IAC 1227 (duro), em solos argiloso e barrento. Em cada experimento foram aplicados dois herbicidas e sua mistura, como segue: I) atrazine a 2,40 kg de i.a./ha, pendimethalm a 1,75 kg e atrazine a 2,00 kg + pendimethalin a 1,25 kg; II) atrazine a 2,40 kg, metolachlor a 3,24 kg e atrazine a 2,00 kg + metolachlor a 2,52 kg; III) cianazine a 2,25 kg, pendimethalin a 1,50 kg e cianazine a 1,75 kg + pendimethalin a 1,00 kg; IV) atrazine a 2,40 kg, alachlor a 2,40 kg e atrazine a 1,60 kg + alachlor a 1,68 kg. Em cada experimento havia um tratamento testemunha. Foram obtidos os pesos de maté ria seca e os comprimentos de raízes e folhas, em quatro épocas, dentro dos períodos iniciais de 36, 49, 27 e 37 dias, respectivamente, para os experimentos I, II, II I e IV. Havia duas repetições para cada trata mento, cada época de amostragem e cada híbrido sendo adotado o delineamento totalmente casualizado. Ós herbicidas pendimethalin e cianazine, isolados ou em mistura, não causaram qualquer efeito no crescimento inicial das plantas de milho. Nos três expe rimentos em que fo i apli cado, o atrazine mostrou efeito estimulante em várias das amostragens, pelo aumento de peso de matéria seca ou de comprimento de raízes. Ó metolachlor e o alachlor apresentaram efeitos fitotóxicos em raízes e folhas, nas primeiras amostragens, com redução de peso e comprimento de folhas e de caules. Dos híbridos estudados, o IAC 7777 e o C-72 aprese ntaram o crescimento mais vigoroso . Ó IAC 1227 apresentou menor tolerância aos herbicidas usados. Ó solo barrento possibilitou melhores condições de crescimentos às plantas de milho que o argiloso, devido à sua fertilidade e textura.

Efeitos dos herbicidas glyphosate e paraquat, aplicados ao solo, sobre a emergência de feijão e soja e de algumas espécies daninhas

Em trabalho conduzido a campo na Estação Experimental Agronômica da UFRGS, Guaíba, RS em 1979/80, usaram-se os herbicidas glyphosate, paraquat e sua combinação, objetivando determinar os efe itos de doses e de épocas de aplicação destes herbicidas em plântulas de feijão e de soja e sobre nutrientes do solo. Pelos resultados obtidos para emergência e peso de maté ria seca da parte aé re a de soja , não houve di ferenças para os tratamentos testados. Também não ocorreram diferenças significativas à população in ic ia l de feijão e sobre os elementos de solo analisados (Ca, Mg e K). Com relação ao peso de matéria seca da parte aérea do feijão, verificou-se que houve interação entre herbicida e doses usadas, tendo glyphosate isolado e glyphosate mais paraquat aplicados seqüencialmente nas doses máximas causado redução significativa naquele parâmetro. Com relação ao paraquat, não foram detectadas diferenças significativas entre as doses testadas.

Estudo de amostragem em experimentos com herbicidas

Tendo como um dos objetivos o estudo de amostragem foi instalado no dia 15 de março de 1981, um experimento no Centro Experimental de Campinas. O Delineamento Experimental utilizado foi em Blocos ao Acaso com três repetições, tendo sido testados nove diferentes herbicidas e mais dois tratamentos testemunha, com e sem cobertura com palha de capim-arroz. Os canteiros, ou unidades experimentais, eram de dimensões 1,OOm x 2,OOm e em todos eles foram semeados alho (Allium sativum L. ) em linhas, espaçadas 0,25m entre si, perfazendo quatro linhas por canteiro. Para fins de avaliação dos efeitos de herbicidas cada canteiro foi dividido em três partes no sentido da linha e em 11 partes no sentido transversal, perfazendo um total de 33 pequenas áreas de dimensões 0,15m x 0,30m que constituiram as unidades básicas para fins de amostragem. Posteriormente, dentro de cada linha os pontos amostrais foram agrupados dois a dois, perfazendo ao todo 15 pontos de dimensões 0,30m x 0,30m, os quais serviram de base para compor as quatro áreas de amostragem indicadas a seguir: X - uma área amostral de dimensões 9,30m x 0,30m centrada no terço médio do canteiro; X2 - uma área amostral centrada no terço médio do canteiro de dimensões 0,30m x 0,90m (X está contido em X2); X3 - uma área amostral centrada no terço médio do canteiro de dimensões 0,30m x 1,50m (X2 está contido em X3); X4 - uma área amostral composto de 3 áreas de dimensões 0,30m x 0,30m tomadas em diagonal, um em cada terço (X1 está contido em X4). A aplicação da Função discriminante aos dados de peso de maté ria seca permitiram tirar as seguintes conclusões: a) para avaliação dos efe itos de hebic idas às plantas daninhas da classe dicotiledôneas as áreas amostrais devem ser centradas no terço médio e correspondente a 4,5% do canteiro; b) para as monocotiledôneas as áreas amostaris devem ser também centradas no terço médio do canteiro e correspondente a 22,5% do mesmo.

Estudo sobre o crescimento de fedegoso

O crescimento de Senna obtusifolia foi estudado sob condições de casa-de-vegetação, num delineamento inteiramente casualizado com 4 repetições . As plantas cresceram em vasos de 6 litros preenchidos com areia e irrigados com solução completa de Hoaglan d, duas vezes por dia. O crescimento das plantas foi avaliado a partir de 21 até 161 dias após a emergência, em intervalos de 14 dias. Os resultados evidenciaram que as plantas de S. obtusifolia acumularam o máximo de matéria seca aos 147 dias após à emergência (D.A.E.) com um período de maior intensidade de crescimento compreendido entre 21 e 63 D.A. E. A partir da emergência até 63 dias, o maior acúmulo de maté ria seca foi verificado nas raízes, após esta data até o final do ciclo do fedegoso, maior proporção foi alocada nos caules. Aos 63 D.A. E. todas as plantas tinham florescido. A taxa de crescimento relativo decresceu rapidamente até 63 dias, mantendo-se baixa e constante até o final do período experimental. Este parâmetro de crescimento foi fortemente influenciado pela tal, a qual foi constante após 91 dias, provavelmente devido ao auto -sombreamento da planta . A área foliar cresceu amplamente até os 35 dias, tornando-se estável a partir dos 119 dias após a emergência das plantas. Considerando-se um período correspondente a dois terços do ciclo médio de cultura s anuais, de dez semanas, S. obtusif olia quando comparada a outras espécies daninhas, apresenta taxas de crescimento inicial menores, o que pode caracterizá-la como uma planta infestante tardia.

Optimizing the width of strip weeding in arabica coffee in relation to crop age

The aim of this study was to determine the weed strip control (WSC) required for adequate coffee growth after transplanting. A non-irrigated, field-planted (spaced 3.80 x 0.70 m) crop was used. The experimental design was a randomized block, with four replicates. The treatments were arranged in a 9 x 18 split-plot design to test the WSC of 0, 15, 30, 45, 60, 90, 120, 150, and 190 cm, which involved continuously hand-weeding at each side of the coffee row, and 18 coffee growth measurements. Multiple regression analyses were carried out relating to growth-variables as a function of both WSC and growth-evaluation times. Brachiaria decumbens was the main weed accomplishing 88.5% of the total weed dry mass. The minimum width of the WSC increases as the crop ages after transplanting. Assuming reductions of 2% and 5% in the maximum coffee growth, the recommended WSC was 75 and 52 cm at 4 months after transplanting (MAT), 104 and 85 cm at 6 MAT, 123 and 105 cm at 9 MAT, 134 and 116 cm at 12 MAT, 142 and 124 cm at 15 MAT, and 148 and 131 cm at 18 MAT, respectively. It was concluded that integrated weed management in young coffee crops must focus on the weed control only in a minimum range along coffee rows, which increases with coffee plant age, keeping natural vegetation in the inter-rows.