Identification of pathogens and virulence profile of Rhodococcus equi and Escherichia coli strains obtained from sand of parks.

The identification of pathogens of viral (Rotavirus, Coronavirus), parasitic (Toxocara spp.) and bacterial (Escherichia coli, Salmonella spp., Rhodococcus equi) origin shed in feces, and the virulence profile of R. equi and E. coli isolates were investigated in 200 samples of sand obtained from 40 parks, located in central region of state of Sao Paulo, Brazil, using different diagnostic methods. From 200 samples analyzed, 23 (11.5%) strains of R. equi were isolated. None of the R. equi isolates showed a virulent (vapA gene) or intermediately virulent (vapB gene) profiles. Sixty-three (31.5%) strains of E. coli were identified. The following genes encoding virulence factors were identified in E. coli: eae, bfp, saa, iucD, papGI, sfa and hly. Phylogenetic classification showed that 63 E. coli isolates belonged to groups B1 (52.4%), A (25.4%) and B2 (22.2%). No E. coli serotype O157:H7 was identified. Eggs of Toxocara sp. were found in three parks and genetic material of bovine Coronavirus was identified in one sample of one park. No Salmonella spp. and Rotavirus isolates were identified in the samples of sand. The presence of R. equi, Toxocara sp, bovine Coronavirus and virulent E. coli isolates in the environment of parks indicates that the sanitary conditions of the sand should be improved in order to reduce the risks of fecal transmission of pathogens of zoonotic potential to humans in these places.

Genetic diversity of Ehrlichia canis in Brazil.

Canine monocytic ehrlichiosis is a highly prevalent disease in Brazil, where the genetic diversity of Ehrlichia canis remains undefined. In this study, we used the TRP36 gene to examine the genetic diversity of E. canis strains from naturally infected dogs residing in five distinct geographic regions in Brazil. E. canis DNA was detected in 82/126 (65%) dogs by dsb-specific PCR and E. canis was isolated in cell culture from 13 dogs. Sequences obtained from dsb genes amplified from the isolates were identical to the US E. canis strain. An extended molecular characterization based on the TRP36 gene identified two major genogroups based on differences among eight isolates. Isolates with tandem repeat amino acid sequence (TEDSVSAPA) identical to the previously reported TRP36 sequence were found in the midwest, northeast and southeast regions of Brazil, and classified into the US genogroup. A novel Brazilian genotype with a different tandem repeat sequence (ASVVPEAE) was also identified in midwest, northern and southern regions. Similarity in the N-terminal sequence of a US genogroup member with the Brazilian genogroup suggested that genomic recombination between the two genogroups may have occurred. Other subtypes within the Brazilian genogroup were also identified using C-terminal amino acid divergence. We identified two distinct major Brazilian genogroups and several subtypes based on analysis of TRP36, and such information will be useful for further genotyping and possible associations with disease severity, understanding of the genetic and antigenic variability of E. canis, and for developing strain-specific vaccines and diagnostic methods based on TRP36.

Identification of pathogens and virulence profile of Rhodococcus equi and Escherichia coli strains obtained from sand of parks

The identification of pathogens of viral (Rotavirus, Coronavirus), parasitic (Toxocara spp.) and bacterial (Escherichia coli, Salmonella spp., Rhodococcus equi) origin shed in feces, and the virulence profile of R. equi and E. coli isolates were investigated in 200 samples of sand obtained from 40 parks, located in central region of state of Sao Paulo, Brazil, using different diagnostic methods. From 200 samples analyzed, 23 (11.5%) strains of R. equi were isolated. None of the R. equi isolates showed a virulent (vapA gene) or intermediately virulent (vapB gene) profiles. Sixty-three (31.5%) strains of E. coli were identified. The following genes encoding virulence factors were identified in E. coli: eae, bfp, saa, iucD, papGI, sfa and hly. Phylogenetic classification showed that 63 E. coli isolates belonged to groups B1 (52.4%), A (25.4%) and B2 (22.2%). No E. coli serotype O157:H7 was identified. Eggs of Toxocara sp. were found in three parks and genetic material of bovine Coronavirus was identified in one sample of one park. No Salmonella spp. and Rotavirus isolates were identified in the samples of sand. The presence of R. equi, Toxocara sp, bovine Coronavirus and virulent E. coli isolates in the environment of parks indicates that the sanitary conditions of the sand should be improved in order to reduce the risks of fecal transmission of pathogens of zoonotic potential to humans in these places.

Recovery threshold of Toxocara canis eggs from soil.

The purpose of this study was to evaluate the threshold of Toxocara canis eggs form soil samples through utilisation of a centrifuge-flotation technique (CFT). Aliquots of soil (1g each) were artificially contaminated with known numbers of T. canis eggs (1, 10, 25, 50, 100, and 200 eggs). The threshold was evaluated based on a CFT using zinc sulphate (Zn(2)SO(4)) and sodium nitrate (Na(2)NO(3)) solutions at a specific gravity of 1.20. The number of eggs recovered was directly proportional to the number of eggs employed to seed the soil. Both solutions enabled full recovery of samples containing merely three eggs; only Zn(2)SO(4) demonstrated efficiency in soil contaminated with a single egg. A recovery rate of 100% was obtained for all tests with samples containing 10 and 25 eggs for Zn(2)SO(4) and Na(2)NO(3), respectively(.) There was no difference in the mean number of recovered eggs regarding either the efficacy of the solutions or the repetition of evaluations in the same trial (p>0.05). Therefore, the CFT is efficient for the detection of Toxocara eggs, even in samples containing low egg numbers.

Toxocariasis/cysticercosis seroprevalence in a long-term rural settlement, São Paulo, Brazil.

Seroprevalence of Toxocara and Taenia solium and risk factors for infection with these parasites were explored in a long-term rural settlement in São Paulo state, Brazil. An ELISA for the detection of anti-Toxocara IgG and IgE and anti-T. solium cysticerci was standardized using Toxocara excretory-secretory antigens (TES) obtained from the cultured second-stage larvae of T. canis and by vesicular fluid antigen from Taenia crassiceps cysticerci (VF). For cysticercosis, the reactive ELISA samples were assayed by Western blot using 18 kDa and 14 kDa proteins purified from VF. Out of 182 subjects, 25 (13.7%) presented anti-Toxocara IgG and a positive correlation between total IgE and the reactive index of specific anti-TES IgE (P=0.0265) was found amongst the subjects found seropositive for anti-Toxocara IgG. In these individuals 38.0% showed ocular manifestations. The frequency of anti-T. solium cysticerci confirmed by Western blot was 0.6%. Seropositivity for Toxocara was correlated with low educational levels and the owning of dogs. Embryonated eggs of Toxocara spp. were found in 43.3% of the analysed areas.

Sero-epidemiology of toxocariasis in a rural settlement in São Paulo state, Brazil.

The seroprevalence of Toxocara canis and risk factors for infection with this parasite were explored in a rural settlement in São Paulo state, Brazil. Total IgA and IgE levels in 79 subjects were determined by turbidimetry and chemiluminescence, respectively. Total counts of leucocytes and erythrocytes and differential counts of leucocytes were made by flow cytometry. ELISA for the detection of anti-Toxocara IgG, IgA and IgE were standardized using Toxocara excretory-secretory antigens (TES) obtained from the cultured second-stage larvae of T. canis. Seventeen (21.5%) of the subjects were found positive for anti-Toxocara IgG, with no significant differences in such seropositivity with age or gender. Thirty (38%) of the subjects showed eosinophilia and 70 (89%) had elevated levels of total IgE. Among the 17 subjects found seropositive for anti-Toxocara IgG, the percentage of leucocytes represented by eosinophils (P=0.0069) and total levels of IgE (P=0.0452) were positively correlated with the levels of anti-TES IgE. Although anti-TES IgA was detected in 10 (59%) of the subjects, there was no significant correlation between the levels of total IgA and those of Toxocara-specific IgA. Only one of the 17 subjects found positive for anti-Toxocara IgG had attended a secondary school and all but two belonged to households with monthly incomes of

Demodectic mange in fattening pigs in São Paulo, Brazil.

An investigation was conducted of mange in five Large White adult sows refractory to pyrethroid treatment (cypermethrin 15%). The most important clinical signs consisted of erythema, hyperkeratosis, alopecic macules, micaceous scaling, follicular pluging, and numerous comedones, especially on the facial region. Skin samples were scraped and punched. Elongate mites were found that were classified as Demodex sp., according to the morphological and histopathological characteristics. The protein, vitamin, and mineral levels contained in the rations were considered to be lower than those recommended to attend the requirements of fattening pigs. Adjustment of the rations was adopted, and the treatment of animals with ivermectin premix (450g/tonne of food) daily orally for a further 7 days was prescribed. The success of treatment was observed 15 days after adoption of these measures, when the beginning of remission of signs was observed. No mites were found on scrapings or biopsy punches after the treatment of the sows.

[Contamination, by Toxocara spp eggs, in public parks and squares in Botucatu, São Paulo, Brazil]. / Contaminação, por ovos de Toxocara spp, de parques e praças públicas de botucatu, São Paulo, Brasil.

The frequency of Toxocara spp eggs in public parks was determined from March 1995 to February 1996. One hundred and twelve samples were collected from 10 public parks in Botucatu, São Paulo State, Brazil. Samples were processed by the decinormal sodium hydroxide concentration method. Out of the 120 soil samples analyzed, 21 were contaminated with Toxocara spp eggs, corresponding to a 17.5% rate of infestation. Of the ten squares submitted to analysis during the year, six presented contamination. Most of the ova found presented characteristics of inviability, with a consequent low chance of human infection, although the population is not free from the risk of contracting this zoonosis.

Contaminação, por ovos de Toxocara spp, de parques e praças públicas de botucatu, São Paulo, Brasil / Contamination, by Toxocara spp eggs, in public parks and squares in Botucatu, São Paulo, Brazil

The frequency of Toxocara spp eggs in public parks was determined from March 1995 to February 1996. One hundred and twelve samples were collected from 10 public parks in Botucatu, São Paulo State, Brazil. Samples were processed by the decinormal sodium hydroxide concentration method. Out of the 120 soil samples analyzed, 21 were contaminated with Toxocara spp eggs, corresponding to a 17.5% rate of infestation. Of the ten squares submitted to analysis during the year, six presented contamination. Most of the ova found presented characteristics of inviability, with a consequent low chance of human infection, although the population is not free from the risk of contracting this zoonosis.

A freqüência de contaminação de parques e praças públicas de Botucatu, São Paulo, Brasil, por ovos de Toxocara spp foi estudada durante 12 meses, com colheitas mensais de amostras de solo de dez praças, que foram processadas pela técnica de concentração em solução decinormal de hidróxido de sódio. Das 120 amostras analisadas, 21 estavam contaminadas, correspondendo a 17,5%, em um total de seis praças. A maioria desses ovos porém apresentaram características de inviabilidade infectiva. Embora a chance de aquisição dessa zoonose nas praças estudadas seja pequena, existe o risco de infecção da população.

Hepatozoon canis infection of wild carnivores in Brazil.

Hepatozoon canis was diagnosed in a crab-eating fox (Cerdocyon thous) found on a highway in the region of Botucatu, São Paulo, Brazil, after being hit by a car. The fox had bilateral fractures of the olecranon, which was corrected by osteosynthesis. Hematologic findings included a neutrophilia, eosinophilia, monocytosis and mild anemia. In the Leishman-stained blood film, gametocytes of Hepatozoon canis in neutrophils were identified measuring 9.1 +/- 0.54 x 5.3 +/- 0.46 microns.