Spatial Distribution of Seropositive Dogs to Leptospira spp. and Evaluation of Leptospirosis Risk Factors Using a Decision Tree

Background: Leptospirosis is an important zoonotic disease associated with poor areas of urban settings of developing countries and early diagnosis and prompt treatment may prevent disease. Although rodents are reportedly considered the main reservoirs of leptospirosis, dogs may develop the disease, may become asymptomatic carriers and may be used as sentinels for disease epidemiology. The use of Geographical Information Systems (GIS) combined with spatial analysis techniques allows the mapping of the disease and the identification and assessment of health risk factors. Besides the use of GIS and spatial analysis, the technique of data mining, decision tree, can provide a great potential to find a pattern in the behavior of the variables that determine the occurrence of leptospirosis. The objective of the present study was to apply Geographical Information Systems and data prospection (decision tree) to evaluate the risk factors for canine leptospirosis in an area of Curitiba, PR.Materials, Methods & Results: The present study was performed on the Vila Pantanal, a urban poor community in the city of Curitiba. A total of 287 dog blood samples were randomly obtained house-by-house in a two-day sampling on January 2010. In addition, a questionnaire was applied to owners at the time of sampling. Geographical coordinates related to each household of tested dog were obtained using a Global Positioning System (GPS) for mapping the spatial distribution of reagent and non-reagent dogs to leptospirosis. For the decision tree, risk factors included results of microagglutination test (MAT) from the serum of dogs, previous disease on the household, contact with rats or other dogs, dog breed, outdoors access, feeding, trash around house or backyard, open sewer proximity and flooding. A total of 189 samples (about 2/3 of overall samples) were randomly selected for the training file and consequent decision rules. The remained 98 samples were used for the testing file. The seroprevalence showed a pattern of spatial distribution that involved all the Pantanal area, without agglomeration of reagent animals. In relation to data mining, from 189 samples used in decision tree, a total of 165 (87.3%) animal samples were correctly classified, generating a Kappa index of 0.413. A total of 154 out of 159 (96.8%) samples were considered non-reagent and were correctly classified and only 5/159 (3.2%) were wrongly identified. on the other hand, only 11 (36.7%) reagent samples were correctly classified, with 19 (63.3%) samples failing diagnosis.Discussion: The spatial distribution that involved all the Pantanal area showed that all the animals in the area are at risk of contamination by Leptospira spp. Although most samples had been classified correctly by the decision tree, a degree of difficulty of separability related to seropositive animals was observed, with only 36.7% of the samples classified correctly. This can occur due to the fact of seronegative animals number is superior to the number of seropositive ones, taking the differences in the pattern of variable behavior. The data mining helped to evaluate the most important risk factors for leptospirosis in an urban poor community of Curitiba. The variables selected by decision tree reflected the important factors about the existence of the disease (default of sewer, presence of rats and rubbish and dogs with free access to street). The analyses showed the multifactorial character of the epidemiology of canine leptospirosis.

Epidemiological aspects in leptospirosis. Research of anti-Leptospira spp antibodies, isolation and biomolecular research in bovines, rodents and workers in rural properties from Botucatu, SP, Brazil.

Leptospirosis is a worldwide infection, transmitted between man and animals that causes a decrease in the production of bovine flocks, and offer risks for public health, as an important zoonosis. The rodents are the main reservoirs of leptospires. It was studied 27 dairy farm properties located in or near from Botucatu-SP, Brazil. In these farms were collected blood and kidney samples from rodents, blood and urine samples from bovines and blood samples from the workers. The serology was performed with microscopic agglutination test (MAT). Samples of bovine urine and rodent kidneys were cultivated searching for leptospires isolation. The polymerase chain reaction (PCR) of the kidneys of the rodents was performed. In MAT, 46/ 140 (32.85%) bovine and 8/34 (23.53%) human sera samples were positive, respectively. In human samples, the serovar Brastilava (37.51%) presented the highest occurrence, while in bovines, the serovars Hardjo and Castellonis were most frequent, with 26.08% each one. All of the rodents were negatives in serology. No leptospire was isolated, and kidney samples were negative in PCR. In bovines, the dam water and the bad hygiene quality of milking process were considered important risks of infection in the affected properties (p<0.05), where other reproductive problems, except abortion, can be related. In other side, to human beings the drainage system was the most important risk factor in the studied properties. Thus, it was verified the necessity of an improvement in zoosanitary handling of the properties, mainly of water supply.

PYOMETRA IN A FEMALE LION (Panthera leo): REPORT of CASE.

Pyometra is a common disease of intact bitches that may also occur in large wild cats. The present study describes a case of pyometra in female lion (Panthera leo). The animal was referred with a vulvar discharge; physical examination and complementary exams did not show abnormalities. An ovariohysterectomy was performed, and the uterus reveals a thick endometrium with purulent/sanguinolent fluid. The bacterial culture resulted in Escherichia coli. The animal had an excellent recovery.

Microbiological evaluation of milk samples positive to California Mastitis Test in dairy buffalo cows (Buballus bubalis)

In order to observe the microbiological status of CMT positive samples, 734 apparently health mammary quarters from buffalo cows were submitted to physical evaluation, strip cup test and CMT. After milk samples inoculation in 10% ovine blood agar base media and in MacConkey agar and incubation under aerobic condition for 72 hours at 37 degrees C, identification was proceeded. According to CMT, 227 quarters (30,93%) were positive, among them 73 (32,16%) presented 1+ reaction, 53 (23,35%) were 2+ and 101 (44,49%) were 3+. Microbiological exams of such samples were positive in 147 (64,76%) out of 227 CMT positive samples and among the remaining 72 (31,72%) were negative and 8 (3,52) were contaminated. In the 147 microbiological positive samples 204 bacteria were found in pure or associated growth and the most frequent agents were: Corynebacterium sp (59,25%); Staphylococcus sp (17,65%) among which 86,11% were coagulase negative and 13,89% were coagulase positive; and Micrococcus sp (6,37%). The results revealed that, excluding the eight contaminated samples, 147 (67,12%) quarters out of 219 CMT positive could be considered as bacteria-carrier and that even in a smaller percentage false-positive results can cause problems in a sanitary program for mastitis control in dairy buffalo cows.

Distribution of contagious and environmental mastitis agents isolated from milk samples collected from clinically health buffalo cows between brazilian dry and rainy seasons of the year

The present study was performed to evaluate the microbiological characteristics of clinically health quarters submitted to milking and also to observe the distribution of contagious and environmental agents between brazilian dry and rainy seasons of the year. During nine months 734 quarters from 37 buffalo cows were submitted monthly to udder inspection, palpation and strip cup test before milking. 734 asseptic milk samples were inoculated in 10% ovine blood agar and in MacConkey agar media, then incubated for 72 hours at 37 C. Among the 580 isolated microrganisms, 182 (31,38%) were recovered from samples collected during the rainy season and 398 (68,62%) from the dry season. In the rainy period the most prevalent agents were: bacteria from the genus Corynebacterium sp (53,30%), Staphylococcus sp (19,78%) and Rhodococcus equi (13,74%). In the dry period, the commonest ones were: Corynebacterium sp (44,97%), Staphylococcus sp (18,84%) and Micrococcus sp (9,55%). The results demonstrated that the methods used to select health quarters in brazilian dairy buffalo farms allow the transmission of contagious bacteria during both seasons of the year, maintaining agents known to cause mainly subclinical inflammatory reactions that compromise cronically the physiology and production of the mammary gland.

Early embryonic death in mares: clinical and hormonal aspects

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estudo do leite de éguas sadias ou portadoras de mastite subclínica, pelo teste de Whiteside, análise microbiológica e contagem de células somáticas

Com o objetivo de identificar a microbiota existente no leite de éguas normais ou portadoras de mastite subclínica, coletaram-se amostras (10 a 15 ml) de leite de 38 animais, que foram examinadas pelo teste de Whiteside, após a homogeneização das amostras dos dois tetos de cada animal. Os resultados foram negativos. Os exames microbiológicos realizados em meios de ágar sangue bovino 10% e ágar MacConkey revelaram presença de bactérias do gênero Staphylococcus spp. e Streptococcus spp. A contagem de células somáticas de amostras individuais de cada teto revelou números superiores a 500.000 células/ml de leite em somente 9,3% das 73 amostras de leite examinadas. Os resultados sugerem a realização de novos estudos objetivando-se a padronização do número de células somáticas no leite de éguas.

Experimental infection of pregnant queens with two major Brazilian clonal lineages of Toxoplasma gondii

Toxoplasma gondii isolates from Brazil are biologically and genetically different from European and North America isolates. Recently, four genotypes were considered the common clonal lineages in Brazil and were designated as types BrI, BrII, BrIII, and BrIV. The pathogenicity of two major Brazilian lineages was investigated after oral inoculation of queens in the middle third of their pregnancies with T. gondii cysts. Twelve pregnant queens without T. gondii antibodies were distributed in group A (infected with a type BrI isolate); group 2 (infected with type BrIII isolate), and group 3 (non-infected control). Infection with type BrI isolate caused toxoplasmosis manifestations and abortion from one litter. Toxoplasmosis manifestations besides premature stillbirth of one litter were observed in queens infected with type BrIII isolate. Indirect fluorescence antibody test showed T. gondii antibodies in all eight infected queens at 30 days after inoculation. In two 10-day-old kittens of the same litter (group 1), titers of 16 and 64 were detected. At the same time, titers of 16, 32, and 32 were detected in three kittens from the same litter (group 2). Experimental infection with tissue cysts from a type BrI and type BrIII isolates of T. gondii developed similar reproductive disturbance in primary infected pregnant queens.

Comparison of conventional PCR, quantitative PCR, bacteriological culture and the Warthin Starry technique to detect Leptospira spp. in kidney and liver samples from naturally infected sheep from Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Prevalence of anti-Toxoplasma gondii antibodies in dairy cattle, dogs, and humans from the Jauru micro-region, Mato Grosso state, Brazil

In order to determine the prevalence of anti-Toxoplasma gondii antibodies, 2000 serum samples from female dairy cattle belonging to 50 farms in the southwest of Mato Grosso state were analyzed by the indirect fluorescent antibody test (IFAT >= 64). Serum samples from 61 dogs (IFAT >= 40) and 116 humans (IFAT = 40), all from the same farm, were also tested. Among these samples, 1420 (71.0%) cattle, 54 (88.5%) dogs, and 113 (97.4%) humans were seropositive for the infection. No significant differences (P >= 0.05) were observed for risk factors associated with the occurrence of toxoplasmosis in humans due to contaminated sources such as fresh milk, cheese/sausage, and contact with felines or other animals. The presence of felines can indicate the likelihood of a contaminated environment, posing a risk to the human population and other animals. The work presented herein is the first report to evaluate the seroprevalence of T gondii in bovines from the Southwest region of the Mato Grosso state, Brazil. (C) 2009 Elsevier B.V. All rights reserved.

Níveis de alumínio e o desenvolvimento de porta-enxertos cítricos em cultivo hidropônico: I. Parâmetros biométricos

O presente trabalho teve como objetivo avaliar a influência de diferentes níveis de alumínio nos parâmetros biométricos dos porta-enxertos cítricos limoeiro Cravo e citrumeleiro Swingle em cultivo hidropônico. O experimento foi instalado em casa de vegetação em Botucatu, SP. Os tratamentos seguiram o delineamento estatístico inteiramente casualizado, com 3 repetições, em parcelas subdivididas. Os níveis de alumínio utilizados foram: 0; 7,5; 15; 22,5 e 30 mg L-1, na forma de AlCl3 6.H2O em solução nutritiva. Foram avaliados os parâmetros biométricos altura, número de folhas, diâmetro do caule, área foliar, comprimento e volume de raízes. de acordo com os resultados obtidos, conclui-se que o citrumeleiro Swingle apresentou-se mais sensível ao alumínio, sendo que, a partir de 7,5 mg L-1 houve redução nos valores destes parâmetros, enquanto que o limoeiro Cravo mostrou-se mais tolerante aos níveis de alumínio.

Níveis de alumínio e acúmulo de macronutrientes em porta-enxertos cítricos em cultivo hidropônico

O presente trabalho teve como objetivo avaliar a influência de diferentes níveis de alumínio no acúmulo de macronutrientes pelos porta-enxertos cítricos limoeiro Cravo e citrumeleiro Swingle, sob cultivo hidropônico. Os tratamentos utilizados em condições de casa de vegetação seguiram o delineamento estatístico inteiramente casualizado, com 3 repetições, em parcelas subdivididas. Os níveis de alumínio utilizados foram: 0, 7,5, 15, 22,5 e 30 mg L-1, na forma de AlCl3.6 H2O. Determinaram-se o acúmulo de N, P, K, Ca, Mg, S e do Al da matéria seca vegetal, relacionando-os com os pesos da matéria seca da parte aérea (folhas e caule) e raízes durante o período de 84 dias, com coletas de plantas a cada 14 dias. Conclui-se que, com adição a partir de 7,5 mg L-1 de alumínio em solução nutritiva ocorreram alterações fisiológicas e reduções significativas no acúmulo de macronutrientes associado à redução nos pesos da matéria seca das folhas, caule e raízes das plantas de citrumelo Swingle.

Influence of different levels of aluminum on the development of citrus rootstock swingle citrumelo (Citrus paradisi mcf. x Poncirus trifoliata raf.) in nutrient solution

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Expressão diferencial dos genes VuUCP1a e VuUCP1b em caupi sob estresse salino

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Study of Toxoplasma infection in Brazilian wild mammals: Serological evidence in Dasypus novemcinctus Linnaeus, 1758 and Euphractus sexcinctus Wagler, 1830

Toxoplasmosis is a worldwide distributed zoonosis that affects man and most warm-blooded animals, with a great economic impact in animal and public health. Serum samples from nine 9-banded armadillos, three 6-banded armadillos, three coatimundis, two opossums and one nutria were submitted for anti-Toxoplasma gondii antibody detection by means of a modified direct agglutination method. Encephalic tissue of three 6-banded armadillos, one 9-banded armadillo, one coatimundi and one nutria were digested in acid pepsin solution and inoculated into Swiss mice for parasite isolation. Only one serum sample from a nine-banded armadillo and two from six-banded armadillos reacted producing titers equal to 256, 512 and 512, respectively. T gondii was isolated in two 6-banded armadillos, one of which was not positive in the serological test. (c) 2005 Elsevier B.V. All rights reserved.