Mast cell and eosinophils in the wall of the gut and eosinophils in the blood stream during Toxocara vitulorum infection of the water buffalo calves (Bubalus bubalis)

Toxocara vitulorum is a pathogenic nematode from the small intestine of very young buffalo calves. To understand the development of the inflammatory responses in the wall of the gut, samples of tissues were removed from the duodenum, jejunum and ileum of buffalo calves naturally infected with T. vitulorum during the beginning of the infection, at the peak of egg output, as well as during the periods of rejection of the worms and post-rejection. Two additional control groups of uninfected calves (by anti-helminthic therapy of their mothers and after the birth) were also necropsied on days 30 and 50 after birth. Blood samples were fortnightly collected from birth to 174 days post-birth. Blood smears were prepared and stained with Giemsa for eosinophils. The parasitological status of buffalo calves was evaluated through weekly fecal egg counts (EPG) from 1 to 106 days after birth, which revealed that T. vitulorum egg shedding started on day 11, reached the peak of the infection on day 49 and finally expelled the parasites between days 50 and 85 after birth. In the infected buffalo calves, the mast cell population increased significantly, by two-fold in the mucosa (villus-crypt unit (VCU)) of the duodenum and four-fold in the proximal jejunum; but these increases were statistically significant only at the peak of the infection. Although mast cell numbers increased in the mucosa of the ileum as well as in both the submucosal and muscle tissues of the duodenum, proximal jejunum and ileum, the data was not significantly different from the controls. Eosinophil numbers increased in the mucosa of the duodenum (two-five times higher than the control) and proximal jejunum (three-five-fold) during the period of the infection (beginning, peak and rejection). The relative numbers of eosinophils increased in the blood stream from the second to the seventh week. In conclusion, T. vitulorum infection elicited mastocytosis and tissue eosinophilia in the duodenum and proximal jejunum, as well as eosinophilia in the blood stream, during the beginning, at the peak and during the rejection of the worm. After the rejection of the worms, the numbers of these cells returned to normal levels suggesting that these cells may have a role in the process of rejection of T. vitulorum by the host. (C) 2003 Elsevier B.V. B.V. All rights reserved.

Eimeria Spp. in Brazilian Water Buffalo

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

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

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

Geographic analysis on the occurrence of human and canine leptospirosis in the city of Maringá, state of Paraná, Brazil

INTRODUÇÃO: Leptospirose é uma zoonose que tem como hospedeiros primários os animais silvestres, sinantrópicos e domésticos. Os humanos comportam-se como hospedeiros terminais e acidentais. Sua prevalência depende dos animais portadores que disseminam o agente, de sua sobrevivência ambiental e do contato de pessoas susceptíveis. Cada sorovar tem um ou mais hospedeiros com diferentes níveis de adaptação. Os focos de leptospirose devem-se aos animais infectados, doentes e assintomáticos, considerados como fontes de infecção ambiental. O objetivo deste estudo foi determinar áreas de risco da infecção leptospírica em cães errantes e pacientes com diagnóstico de leptospirose nos anos de 2006 a 2008, em Maringá, Estado do Paraná, Brasil. MÉTODOS: Foram estudados 335 cães errantes e 25 pacientes. Os soros, tanto dos animais como dos pacientes, foram examinados pela prova de soroaglutinação microscópica (SAM), para pesquisa de anticorpos antileptospíricos. Para determinar áreas de risco e a distribuição espacial da doença foram elaborados mapas temáticos. RESULTADOS: Foram observados 41(12,2%) cães positivos para um ou mais sorovares de leptospiras, e os mais frequentes foram: Pyrogenes (43,9%), Canícola (21,9%) e Copennhageni (19,5%). Nos humanos, a positividade foi de 2 (8%) para os sorovares, Pyrogenes e Hardjo Prajitno e, Pyrogenes e Cynopteri. CONCLUSÕES: A análise espacial revelou que o risco de cães e humanos, no município de Maringá, se infectar com leptospiras está presente tanto em áreas centrais como periféricas, fato que reforça a relevância deste estudo e de ações contínuas de vigilância epidemiológica e ambiental para o controle da doença tanto nos animais como no homem.

A nonsense mutation in the tyrosinase gene causes albinism in water buffalo

Background: Oculocutaneous albinism (OCA) is an autosomal recessive hereditary pigmentation disorder affecting humans and several other animal species. Oculocutaneous albinism was studied in a herd of Murrah buffalo to determine the clinical presentation and genetic basis of albinism in this species.Results: Clinical examinations and pedigree analysis were performed in an affected herd, and wild-type and OCA tyrosinase mRNA sequences were obtained. The main clinical findings were photophobia and a lack of pigmentation of the hair, skin, horns, hooves, mucosa, and iris. The results of segregation analysis suggest that this disease is acquired through recessive inheritance. In the OCA buffalo, a single-base substitution was detected at nucleotide 1,431 (G to A), which leads to the conversion of tryptophan into a stop codon at residue 477.Conclusion: This premature stop codon produces an inactive protein, which is responsible for the OCA buffalo phenotype. These findings will be useful for future studies of albinism in buffalo and as a possible model to study diseases caused by a premature stop codon.

Perfil de aglutininas anti-Leptospira em bezerras búfalas vacinadas com bacterina pentavalente comercial contra leptospirose

Avaliou-se o perfil sorológico de 17 bezerras búfalas vacinadas nos dias zero, 30 e 210 com bacterina pentavalente comercial contra leptospirose, empregando-se a prova de soroaglutinação microscópica com antígenos vivos, durante o período de 360 dias. Utilizaram-se quatro bezerras-controle, e todos os animais foram negativos no dia zero. Nos animais vacinados, foram observadas reações aos 60 dias para todos os sorovares, associadas a títulos máximos para Canicola, Grippothyphosa e Pomona. No 90° dia, houve declínio acentuado dos títulos para todos os sorovares e ausência de títulos para Hardjo. Nos dias 120º, 150º, 180º e 210º, ocorreram oscilações de títulos para todos os sorovares. No 240º dia, foram encontrados títulos máximos para Hardjo, elevação para os outros sorovares e ausência de título para Canicola. Nos dias 270º, 300º e 330º, foi observado declínio gradativo dos títulos. No 360º dia, foram encontrados títulos residuais para Icterohaemorrhagiae, Grippothyphosa e Pomona e ausência de reação para os outros sorovares. Inferiram-se a reduzida persistência dos títulos vacinais mesmo após o reforço e revacinação e a baixa interferência da vacinação no diagnóstico sorológico.

Occurrence and risk factors associated with canine leptospirosis

Leptospirosis is a globally distributed emerging zoonosis. Dogs are commonly affected and although other serovars can cause canine leptospirosis, Leptospira interrogans serovar Canicola is primary found in these animals. A retrospective study was conducted using a database of 1195 dogs tested for Leptospira infection from 2003 to 2010 at the Laboratory of Zoonosis Diagnosis at the Veterinary Hospital of São Paulo State University (UNESP) in Botucatu, São Paulo state, Brazil. The seroprevalence of infected dogs was 20.08% (240/1195), and the most prevalent serovars were Canicola (6.7%), Copenhageni (5.0%), Icterohaemorrhagiae (2.9%), Autumnalis (2.9%), Pyrogenes (2.8%), Pomona (2.0%), Hardjo (2.0%), Australis (1.8%), Bratislava (1.6%), Cynopteri (1.4%), Grippotyphosa (1.3%) and Djasiman (1.0%). By univariate analysis, the variables age and breed were not statistically related to the infection, while gender and season were. The effects of gender were also noticeable related to serovars Australis, Canicola and Hardjo. In multivariate analysis, the level of significance (p-value) of season was suppressed by gender, indicating possible collinearity between those two variables.

Interactions between environment, wild animals and human leptospirosis

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

An in vitro growth inhibition test for measuring the potency of Leptospira spp. Sejroe group vaccine in buffaloes

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

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.

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.

Parturition intervals and distributions of parturitions by months of buffalo in Southern and South-eastern Brazil

Reproductive rate is an important component of economic success in livestock production. Parturition interval (IEP) is a direct measure of the productivity of the animal. Long IEP reduce the number of calves produced per year. The objective this study was to determine the distribution of parturitions across month and to evaluate factors affecting IEP. The data included 7,588 parturitions of Murrah, Mediterranean and Carabobo buffalo from 10 herds in Southern and South-eastern Brazil. The analysis of distribution of parturitions evaluated the effects of month, year and their interaction on birth date of calves by using a Chi-Square test in SAS PROC FREQ (SAS Institute, Cary, NC, USA). Parturition intervals (n = 2,630) were evaluated using analysis of variance in SAS PROC GLM. The model for IEP included the fixed effects of season (December to May = 1, June to November = 2), year, season x year, sex of the preceding parturition, age of weaning of the previous calf, and herd. All sources of variation were significant (P<0.0001) except sex of the preceding parturition (P <0.85). The mean IEP was 446.7 +/- 10.4 days, for seasons 1 and 2 IEP were 419.8 +/- 11.3 and 473.6 +/- 40.7 days, respectively, a difference of 54 days. As weaning age increased there was a lengthening of IEP. Buffalo in Brazil showed seasonal parturition with calving concentrated from January to April, although the frequency by month differed across years (P<0.0001). These months also had the lowest calving interval.

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.

Evaluation of water beef buffalo from birth to two years using different growth curves

The buffalo is a domestic animal species of growing world-wide importance. Research to improve genetic improvement programs is important to maintain the productivity of buffalo. The objective this research was to evaluate the growth of Brazilian buffalo to two years of age with different growth curves. Growth curves consolidate the information contained in the weight-age data into three or four biologically meaningful parameters. The data included 31,452 weights at birth and 120, 205, 365, 550 and 730 days of buffalo (n = 5,178) raised on pasture without supplementation. Logistic, Gompertz, quadratic logarithmic, and linear hyperbolic curves (designated L, G, QL, and LH, respectively) were fitted to the data by using proc NUN of SAS (SAS Institute, Inc., Cary, NC, USA). The parameters estimates for L [WT= A * (((1 + exp (-k * AGE)))**-m)] were A = 865.1 +/- 5.42; k= 0.0028 +/- 0.00002; M= 3.808 +/- 0.007; R(2) = 0.95. For G [WT= A * exp (-b * exp (-k * age)] the parameters estimates were A= 967.6 +/- 7.23; k = 0.00217 +/- 0.000015; b = -2.8152 +/- 0.00532. For QL [WT= A + b*age + k*(age*age) + m*log (age)] parameters estimates were A= 37.41 +/- 0.48; k= 0.00019 +/- 6.4E(-6); b= 0.539 +/- 0.006; m= 2.32 +/- 0.23; R(2)=0.96. For LH [WT= A + b*AGE + k*(1/AGE)] the parameters estimates were A= 23.15 +/- 0.44; k=15.16 +/- 0.66; b= 0.707 +/- 0.001; R(2)= 0.96. Each of these curves fit these data equally well and could be used for characterizing growth to two years in beef buffalo.