First isolation and molecular characterization of Toxoplasma gondii from finishing pigs from São Paulo State, Brazil

Toxoplasma gondii infection is widely prevalent in humans in Brazil. Among the food animals, pigs are considered the most important meat source of T. gondii for infection in humans. In the present study, we report the first isolation of viable T. gondii from finishing pigs in Brazil. Antibodies to T. gondii were found in 49 (17%) of 286 pigs prior slaughter using the modified agglutination test (MAT) at a serum dilution of 1:25. Attempts were made to isolate T. gondii from 28 seropositive pigs. Samples of heart, brain, and tongue from each pig were pooled, digested in acid pepsin, and bioassayed in five mice per pig. Viable T. gondii was isolated from seven pigs; all isolates were lethal for mice. Restriction fragment length polymorphism on products of SAG2 locus amplified by PCR revealed that two isolates were Type I and five were Type III. The results indicate that phenotypically and genetically T. gondii isolates from pigs from Brazil are distinct from isolates of T. gondii from pigs in the USA.     

Serological profile of Toxoplasma gondii and Neospora caninum infection in commercial sheep from São Paulo State, Brazil

Neosporosis and toxoplasmosis are two important infections in young and adult sheep, leading to low production and abortion. This study aimed to determine the frequency of antibodies to Toxoplasma gondii and Neospora caninum in sheep from the eastern region of S?o Paulo State, Brazil. Serum samples (382) were collected from the sheep and assayed for T. gondii through modified agglutination test (MAT) and indirect fluorescent antibody test (IFAT), and for N. caninum antibodies, through IFAT, with cut-off titers equal to 16 (T. gondii) and 25 (N. caninum). All frozen samples were sent to the Center for Zoonoses Research (NUPEZO), Department of Veterinary Hygiene and Public Health (DHSVP), FMVZ, UNESP, for serological tests. A total of 71/382 (18.6%) samples reacted to T. gondii, especially at titers 16 (28; 39.4%), 64 (15; 21.1%), 256 (21; 29.6%) and 1024 (6; 8.5%) by MAT, and 16 (34; 47.9%), 64 (18; 25.4%), 256 (14; 19.7%) and 1024 (5; 7%) by IFAT. As regards N. caninum, 49/382 (12.8%) samples reacted at titers 25 (17; 34.7%), 50 (11; 22.5%), 100 (11; 22.5%), and ≥ 200 (10; 20.4%). These animals presented infection but no clinical signs. Six and ten animals had high titers for toxoplasmosis and neosporosis. No significant association was observed between antibodies for both parasites (P=0.535) according to Fisher's exact test, and no correlation was found between T. gondii (MAT) and N. caninum antibody titers (r=-0.0068; P=0.895), T. gondii (IFAT) and N. caninum antibody titers (r=-0.0025; P=0.961). Thus, T. gondii and N. caninum infections were observed in farms located in S?o Paulo State, where sheep play an important economical role for the national and regional business.     

Rabies seroprevalence of street dogs from São Paulo City, Brazil

A sample of 602 street dogs captured by the São Paulo zoonosis-control municipal service was analyzed for the presence of serum antibodies against the rabies virus and compared to a control sample of 37 dogs from the kennel of the police of São Paulo. Data were stratified into 4 age intervals and the age from the street-dog sample was estimated by general aspects and dental observation according to standard techniques. We assumed that seropositivity was due to previous vaccination (no sub-clinical infection). From the prevalence data, we estimated the age-dependent incidence-density rate of vaccination by a simple mathematical model, which also allowed the estimation of the period of antibody protection given by the supposed vaccination. Crude seroprevalence in the street-dog sample was 16.5% and in the sample from the police-kennel dogs was 80%. The average age of first vaccination calculated for the street-dog sample was 5 years of age (if we assumed an average period of vaccine protection of 3 years) — but 5 months of age for the police-kennel sample.     

Multidrug and multispecies resistance in sheep flocks from São Paulo state, Brazil

The economic importance of sheep production is increasing worldwide simultaneously with the emergence of parasitic resistance. This study aimed to survey the current situation of management practices and parasite resistance in sheep flocks in S?o Paulo state, Brazil. A questionnaire was given to 35 sheep farmers to obtain information related to flock management practices. Of these flocks, 30 were submitted to the fecal egg count reduction test (FECRT) with at least one of the five following anthelmintics: albendazole, closantel, ivermectin, levamisole, and moxidectin, for comparison against an untreated control group. In the survey, the median number animals per flock was 301, mainly of the Santa Ines breed (in 75.8% of the flocks) and crossbred animals (in 54.5% of the flocks). The predominant farming system was semi-intensive (82.9%), using rotational grazing (80%). Selective treatment was based on FAMACHA grade (47.1%) and in clinical signs (41.2%). The most often applied anthelmintics were macrocyclic lactones (42.9-54.2% in the last three applications). Considering the anthelmintics employed in this study, 10.7% of the farms' flocks were resistant to three, 35.7% to four, and 53.6% to all five anthelmintics. The main helminth genera observed before and after treatments were Haemonchus sp. (75.8%) and Trichostrongylus sp. (19.1%), but all observed genera (Cooperia sp., Oesophagostomum sp., and Strongyloides sp.) were detected by the FECRT. Considering efficacy values less than or equal to 90% in the FECRT as resistant, 100% of flocks were resistant to albendazole and ivermectin, 96.6% to moxidectin, 92.9% to closantel, and 53.6% to levamisole. It is thus possible to conclude that multidrug resistance is widespread in sheep flocks in S?o Paulo state, Brazil, and this involves all prevalent helminth genera.     

Prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in ovine from São Paulo State, Brazil

Serum samples from 597 sheep from S?o Paulo State, in the southeastern region of Brazil, were tested to determine the prevalence of antibodies directed against Toxoplasma gondii (> or = 1:64) and Neospora caninum (> or = 1:50) using the indirect fluorescent antibody test (IFAT). The animals were divided into three groups based on their age: < or = 1 year, 1-4 years, and > or = 4 years. Antibodies to T. gondii were observed in 34.7% of the samples with titers ranging from 64 to 16,384 and IgG antibodies directed against N. caninum were observed in 9.2%, with titers ranging from 50 to 3200. Only 3.5% of the sheep were positive for both agents. All farms had at least one positive animal for T. gondii, and 26 of the 30 farms had at least one positive animal for N. caninum. An association between seroprevalence and age was observed for T. gondii (P = 0.001), but not to N. caninum (P = 0.343). It was not possible to associate seroprevalence to T. gondii and the presence of domestic or feral cats, since in all farms there was at least one positive sheep. There was no association between seropositivity to N. caninum and the presence of domestic (P = 1.000) and feral dogs (P = 0.550).     

Leptospirosis seroprevalence and risk factors for sheep in Maranhão state,Brazil

This study was conducted to determine leptospirosis seroprevalence in sheep and their spatial distribution as well as identify risk factors associated with seropositivity in sheep from 37 herds and 11 municipalities in the Presidente Dutra microregion, Maranhão state, Brazil. We analyzed 379 blood serum samples using a Microscopic Agglutination Test (MAT). The individual seroprevalence was 32 %. Of the 37 herds studied, 30 (81 %, 95 % CI 69–94 %) had at least one seropositive animal. In seven municipalities, we observed infection in 100 % of the herds. The serovars recorded were Grippotyphosa (67 %), Wollfi with Hardjo (9 %), Bratislava (9 %), Hardjo (5 %), Icterohaemorrhagiae (5 %), Pomona (2 %), Castellonis (2 %) and Copenhageni (0.8 %). We concluded that the Leptospira spp. in sheep is widespread in the area of sheep farms in Maranhão state, and a risk factor is the animals' water source.     

Seroprevalence and isolation of Toxoplasma gondii from free-range chickens from Espírito Santo state, southeastern Brazil

Prevalence of Toxoplasma gondii infection in 510 free-range (FR) chickens (380 from 33 small farms, and 130 from a slaughter house for FR chickens) from Espírito Santo state, southeastern Brazil, was investigated. Antibodies to T. gondii were sought using commercial indirect haemagglutination (IHAT, Imuno-HAI Toxo(?), Wama Diagnóstica, S?o Paulo, Brazil, cut-off 1:16) and the modified agglutination test (MAT, cut-off 1:25) tests. Attempts were made to isolate viable T. gondii from seropositive chickens by bioassay in mice. Pooled samples of brain, heart and quadriceps muscle of one thigh (total 40 g) from 64 chickens with IHAT titers of ≥ 1:16 were minced, digested in pepsin and bioassayed in mice. Antibodies to T. gondii were found in 40.4% (206/510) FR chickens by IHAT (titer ≥ 1:16) and 38.8% (198/510) by MAT (titer ≥ 1:25); concordance between IHAT and MAT was 81.6% (kappa index=0.614). Viable T. gondii was isolated (designated TgCkBr234-281) from 48 of 64 (75%) seropositive (IHAT titers ≥ 1:32) FR chickens. Most isolates of T. gondii were virulent for mice; 100% of mice inoculated with 44 of 48 isolates died of toxoplasmosis within 30 days post inoculation (p.i). An epidemiological investigation revealed that people living in rural areas have little knowledge about the parasite and about the risk of acquiring it from raw meat. Results indicated that the locally available IHAT was useful for screening of chicken sera for T. gondii antibodies.     

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.     

Molecular detection of Leishmania sp. in cats (Felis catus) from Andradina Municipality, São Paulo State, Brazil

The aim of this work was to molecularly detect Leishmania species in 52 cats from Andradina Municipality, S?o Paulo State, Brazil. The direct parasitological test was performed by using imprints of poplited lymph node, bone marrow and spleen to verify amastigote forms of Leishmania spp. The samples that were positive parasitological tests were subjected to molecular analysis (PCR) and sequencing. Infection was detected for 5.76% (3/52) of the examined cats and two had presence of amastigote forms of Leishmania spp. in lymph nodes. Polymerase chain reaction (PCR) of kinetoplast minicircle DNA, indicated positive amplification for samples of spleen and lymph nodes and the sequencing resulted in 97% similarity with Leishmania (L.) chagasi. This study proved the occurrence of infection with Leishmania (L.) chagasi in felines from Andradina municipality, S?o Paulo State.     

Anthelmintic resistance in cattle nematodes in northwestern São Paulo State, Brazil

The present study was carried out in the northwestern region of S?o Paulo State, Brazil, to determine the anthelmintic resistance status in cattle naturally infected with gastrointestinal nematodes. The anthelmintics tested were levamisole phosphate (Ripercol, Fort Dodge), albendazole sulphoxide (Ricobendazole, Fort Dodge), ivermectin (Ivomec, Merial) and moxidectin (Cydectin, Fort Dodge), administered at the doses recommended by the manufacturers. From April 2002 to May 2004, 25 cattle farms were evaluated. On each farm, steers were divided into treatment and control (not treated) groups based on fecal egg counts (FEC). Between 7 and 10 days after the anthelmintics administration, fecal samples were collected from each animal for post-treatment FEC. Fecal cultures from each group were also prepared for larval identification. After treatment, mean FEC reduction (FECR) in treatment groups (compared with control groups) was assessed on each farm. FECR was lower than 90% on 23 farms after ivermectin treatment. On 19 farms, FECR of 100% was recorded following moxidectin treatment; on the remaining 6, FECR ranged from 90% to 97.2%. After albendazole treatment, FECR was higher than 90% on 20 farms and ranged from 47.4% to 84.6% on other 5. After levamisole treatment, FECR was higher than 90% on 23 farms and equal to 47.4% and 73.7% on other 2 farms. Results indicated the presence of resistant Cooperia spp. and Haemonchus spp., especially to ivermectin; on some farms, resistance to albendazole and levamisole was also observed.     

Risk factors to tick infestations and their occurrence on horses in the state of São Paulo, Brazil

From December 1998 to March 1999, 40 stud farms were studied in the state of S?o Paulo, Brazil. During visits to farms, horses reared under grazing conditions were examined for the presence of ticks. On each farm visit, horse pastures were closely inspected and a questionnaire was given to the farm supervisor with the purpose of gaining information about ecological and management variables (independent variables) that could be associated with the presence and infestation levels of ticks on the farm (dependent variables). Three tick species were found during the study. Anocentor nitens, Amblyomma cajennense and Boophilus microplus were present on horses from 38 (95%), 20 (50%) and four (10%) farms, respectively. All farms that had A. cajennense or B. microplus infestations also had A. nitens infestations. Only one of the four farms with B. microplus infestations on the horses also had A. cajennense infestations. Two farms had all horses free of ticks. There was a strong association between the presence of infestation by B. microplus on horses and the simultaneous use of a grazing area by cattle and horses (P = 0.000). There was no statistical association between any of the independent variables and the presence or infestation level of A. nitens on the horses (P > 0.20). The presence of A. cajennense was statistically associated with the presence of at least one mixed overgrowth pasture in the farm (P = 0.001). A mixed overgrowth pasture means the presence of undesired plants such as bushes and shrubs in the pasture. The presence of high levels of A. cajennense on horses was also associated with the presence of at least one mixed overgrowth pasture in the farm (P = 0.026). The regular use of acaricides was statistically associated with the presence of ticks on the horses (P < 0.05), making this procedure a result of the inefficacy of controlling ticks on the farms. The occurrence of human infestation by ticks was statistically associated with the presence of A. cajennense on the horses (P=0.000). The presence of at least one mixed overgrowth pasture on the farm was associated (P = 0.000) to either higher horse densities and to farms that did not mow all the pastures once a year, indicating that mowing all the pastures at least once a year can be considered a protective factor against the presence of mixed overgrowth pastures on the farm, and consequently, against the presence of A. cajennense on the horses.     

Molecular identification of Cryptosporidium spp. from fecal samples of felines, canines and bovines in the state of São Paulo, Brazil

The objective of this study was to obtain information of epidemiological nature through genotypic characterization of Cryptosporidium isolates from dogs, cats and bovines from the state of São Paulo, Brazil. The extraction of DNA from oocysts was carried out and polymerase chain reaction was accomplished using specific primers to 18S rRNA gene. The amplicons were directed sequenced. Seven cat samples, nine dog samples and nine bovine samples were analysed. From the seven cat samples the genotypic analyses revealed Cryptosporidium felis in all. These were the first genotypic characterization of Cryptosporidium from domestic felines in Brazil. In nine sequenced samples from dogs, genotypic identities compatible with Cryptosporidium canis were revealed in all samples. The genotypic analyses in bovines revealed Cryptosporidium parvum in eight samples and Cryptosporidium bovis in another sample, the last one being a non-zoonotic species, not related to clinical symptoms and described for the first time in Brazil.     

Occurrence and molecular diagnosis of Cryptosporidium serpentis in captive snakes in S?o Paulo, Brazil

The present study aimed to determine whether Cryptosporidium oocysts were present in stools from captive snakes at Funda??o Parque Zoológico (Zoological Park Foundation) in S?o Paulo, Brazil. Two collections were performed; the first in July 2008 and the second in February 2009. Fecal samples were collected from 74 enclosures that housed 101 individuals of 23 snake species. The stool specimens collected from 16 out of the 74 enclosures (21.6%) contained Cryptosporidium spp. oocysts; all of them were confirmed as Cryptosporidium serpentis, using molecular techniques. Only in three (18.7%) out of the 16 enclosures with positive samples were there animals with clinical signs compatible with infection by C. serpentis, such as regurgitation and significant progressive weight loss. From the results, it was concluded that diagnostic examinations need to be performed periodically, even on clinically healthy animals, as a preventive measure.     

Seasonal dynamics of ticks (Acari: Ixodidae) on horses in the state of São Paulo, Brazil

Natural tick infestations were assessed every 14 days on horses over a 2-year period. Amblyomma cajennense adult ticks were counted individually, without detachment from the horses. Larvae and nymphs of A. cajennense were collected using a rubber scraper that scratched engorged immature ticks from the host. Adult females of Anocentor nitens larger than 4mm length were counted on the horses. Blood samples were also obtained from the horses every 14 days and macroclimatic data were obtained for the study period. Infestations of A. cajennense demonstrated distinct peaks of activity for each of the three parasitic stages over each 12-month period, showing a 1-year generation pattern. Larvae predominated from April to July and nymphs from June to October. Adults predominated from October to March with a greater number of adult males than females. Although other studies on seasonal dynamics in the states of Rio de Janeiro and Minas Gerais were performed with the free-living stages of A. cajennense on pastures, the present study in the state of S?o Paulo, performed with the parasitic stages of A. cajennense on horses, showed similar results to those observed in other states. Infestations by A. nitens demonstrated distinct peaks of activity of adult females (>4 mm), suggesting different tick generations during the year. Infestation with A. nitens was much higher in the first year than the second year which may have been related to horse nutritional status and stocking rate. Although several climatic variables showed statistical significant correlation (r) with tick counts, the determination coefficients (R(2)) were always lower than 0.40, suggesting that any single significant variable (i.e. mean temperature) would not explain the tick distribution pattern over the year. The highest peaks of A. nitens females (>4 mm) were significantly associated with decrease in horse packed cell volumes (R(2)=0.603). The ears and the perineum, tail and groin region accounted for around 70% of all A. nitens females counted on the horses.     

Prevalence of dog intestinal parasites and risk perception of zoonotic infection by dog owners in São Paulo State, Brazil

Coprological examination was used to estimate the prevalence of gastrointestinal parasites in stray and domiciled dogs from Botucatu, S?o Paulo State, Brazil. Risk factors for dog infection were assessed in relation to demographic, husbandry and management data. The dog owners completed a questionnaire survey on some aspects of dog parasitism such as parasite species, mechanisms of infection, awareness of zoonotic diseases and history of anthelmintic usage. Parasites were found in the faeces of 138 dogs, with an overall prevalence of 54.3%. Dogs harbouring one parasite were more common (31.4%) than those harbouring two (18.5%), three (3.2%) or four (1.2%). The following parasites and their respective frequencies were detected: Ancylostoma (37.8%), Giardia (16.9%), Toxocara canis (8.7%), Trichuris vulpis (7.1%), Dipylidium caninum (2.4%), Isospora (3.5%), Cryptosporidium (3.1%) and Sarcocystis (2.7%). Stray dogs were found more likely to be poliparasitized (P<0.01) and presented higher prevalence of Ancylostoma, T. canis and Giardia (P<0.01) than domiciled ones. Toxocara canis was detected more frequently in dogs with <6 months of age (P<0.05) and no effect of sex or breed could be observed (P>0.05). Except for Ancylostoma, that showed a significantly higher prevalence in dogs living in a multi-dog household (P<0.01), parasite prevalences were similar in single- and multi-dog household. The answers of dog owners to the questionnaire showed that the majority does not know the species of dog intestinal parasites, the mechanisms of transmission, the risk factors for zoonotic infections, and specific prophylactic measures. The predominance of zoonotic species in dogs in the studied region, associated with the elevated degree of misinformation of the owners, indicates that the risk of zoonotic infection by canine intestinal parasite may be high, even in one of the most developed regions of Brazil.     

Occurrence of Neospora caninum antibodies in sera from dogs of the city of São Paulo,Brazil

Neospora caninum is an important cause of abortion in dairy cattle worldwide. Dogs are important in the epidemiology of this parasite because they are the only hosts known to excrete N. caninum oocysts. In order to understand the prevalence of N. caninum in dogs, sera from 500 owned dogs and from over 600 feral street dogs from the city of S?o Paulo, Brazil were assayed for antibodies to N. caninum. Sera were examined by the Neospora agglutination test (NAT) using mouse-derived tachyzoites. Antibodies (> or =1:25) to N. caninum were found in nearly 10% (49/500) of owned dogs and in 25% (151/611) of stray dogs. NAT titers for owned dogs were 1:25 in 28 (5.6%) dogs, 1:50 in 20 (4%) dogs, and > or =1:500 in 1 (0.2%) dog. NAT titers for stray dogs were 1:25 in 79 (12.9%) dogs, 1:50 in 68 (11.1%) dogs, and > or =1:500 in 4 (0.6%) dogs. These data indicate that feral dogs may be important in the epidemiology of N. caninum infection.     

Seroepidemiology of Toxoplasma gondii and Neospora caninum in dogs from the state of Paraíba, Northeast region of Brazil

A cross-sectional study was conducted to determine the seroprevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies and to investigate the risk factors related to antibodies against T. gondii and N. caninum in dogs of the city of Campina Grande, state of Paraiba, Northeast region of Brazil. For this purpose, 286 blood samples were collected from dogs during the rabies vaccination campaign, in September 2003, and on this occasion questionnaires addressing epidemiological aspects of the infections were given to each dog owner. The sera were analyzed for anti-T. gondii and anti-N. caninum antibodies by indirect fluorescent antibody tests. Of the total of 286 dogs, 129 were positive for T. gondii (titer16) with a prevalence value of 45.1% (95% CI=39.24-51.07%). For N. caninum, 24 dogs were positive (titer50), with a prevalence value of 8.4% (95% CI=5.45-12.23%). Antibodies to T. gondii and N. caninum were found simultaneously in 14 dogs (4.9%; 95% CI=2.7-8.08%). For T. gondii infection, the risk factors associated with seroprevalence was the age of the animals, with dogs older than one year presenting higher values of odds ratio, and co-habitation of cats in the household. For N. caninum infection, dogs that have street contact had higher odds of seropositivity than dogs that remained exclusively in a domestic environment.     

A cross-sectional study of swine influenza in intensive and extensive farms in the northeastern region of the state of São Paulo,Brazil

Swine influenza (SI) is a seasonal infectious disease highly important to the world pig industry. Loss of daily weight gain, increased costs for the prevention and treatment of secondary infections are the main economic losses associated with the presence of this disease. However, some epidemiological features of SI remain quite unclear. This study focused on assessing the prevalence of swine influenza virus (SIV) infection in intensive and extensive pig herds and associating risk factors. A set of 601 blood samples of five intensive farrow-to-finish farms and 361 blood samples from 56 extensive farms were analyzed using an indirect ELISA kit CIVTEST SUIS INFLUENZA®, Hipra (Amer, Spain), in order to detect anti-SIV antibodies. In total, 24.13 % of samples from intensive herds were positive, while no positive samples were detected in extensive rearing herds. Sow and weaning piglets had the highest prevalence values. In the intensive rearing system, occurrence of reproductive disorders and exposure to recently introduced animals were positively associated with the disease occurrence in swine herds. The findings highlight the importance of sows in the epidemiology of the disease and bring information about risk factors involved in the occurrence of swine influenza in intensive herds.     

Prevalence of Neospora caninum and Toxoplasma gondii in sheep and dogs from Guarapuava farms, Paraná State, Brazil

Sheep and dog blood samples were collected from nine farms in the county of Guarapuava, Paraná, Brazil. The indirect fluorescent antibody test (IFAT) was used to detect Neospora caninum and Toxoplasma gondii antibodies. Herein, serum samples from 305 sheep were evaluated, being 29 (9.5%) and 157 (51.5%) seropositives to N. caninum and T. gondii, respectively. Seven (29.1%) and five (20.8%) out of 24 dogs were seropositives to N. caninum and T. gondii, respectively. There were no differences among the sheep serology for N. caninum and reproductive problems, management and animal feeding variables, neurological problems and presence of other animals species on the farm (P>or=0.05). The simultaneous frequency of antibodies between N. caninum and T. gondii was 5.2% in the herds. Age, breed, farm size, semi-intensive activity, mineral salt supplementation, water origin, stage of the pregnancy when reproduction problems occurred, neurological problems in lambs, presence of rodents in the food room and pasture cat access were identified as associated factors for the occurrence of toxoplasmosis in sheep (P<0.05). There were no differences among the seropositivity in dogs for N. caninum and T. gondii and breed, age and sex (P>or=0.05). The present work is the first report on serum prevalence of N. caninum in sheep from the state of Paraná, Brazil.     

An investigation of Leishmania spp. in Didelphis spp. from urban and peri-urban areas in Bauru (São Paulo, Brazil)

Blood and bone marrow samples were taken from 112 Didelphis spp., collected between March 2005 and February 2006, from urban and peri-urban areas of Bauru, São Paulo State, Brazil, to evaluate the hypothesis that these animals might constitute a reservoir of Leishmania spp. Anti-Leishmania ssp. antibodies were screened in the serum samples using an enzyme-linked immuno-sorbent assay (ELISA) and the polymerase chain reaction (PCR). PCR was performed on fragments of DNA samples from Leishmania spp. using primers 13A and 13B, and showed a positive outcome in 91.6% of the 112 samples tested. Of the 107 samples analyzed by ELISA, 71% were positive. Evidence of epidemiological risk factors such as a circulating parasite and freely moving vectors suggests that Didelphis spp. may participate in the transmission cycle of Leishmania spp. in Bauru.