Dayana Firmino de Morais

Toxoplasma gondii and Neospora caninum in slaughtered pigs from Northeast, Brazil

The aims of this study were to evaluate the presence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies, to identify associated risk factors and to isolate T. gondii from slaughtered pigs in Paraíba State, Brazil. A total of 190 pigs from public slaughterhouses were used in the study. An indirect immunofluorescence test was used to detect antibodies, and the isolation of T. gondii was performed with a bioassay in mice based on tissues from seropositive animals. A total of 50 g of brain, heart and tongue tissue from 37 positive pigs with titres ≥ 1:64 was ground, digested with acidic pepsin and inoculated into mice. The prevalence of anti-T. gondii antibodies was 19.5% (95% CI: 14.1%; 25.8%) (37/190), with titres ranging from 1:64 to 1:2048. Viable T. gondii parasites were isolated from 13 of the 37 (35.1%; 95% CI: 20.2%; 52.5%) seropositive pigs. We found that the virulence of the isolates varied; three of these isolates were able to kill all of the inoculated mice. The risk factors for infection were extensive husbandry and feeding with leftovers. The prevalence of antibodies to N. caninum in pigs was 3.2% (95% CI: 1.2%; 6.7%) (6/190), with titres ranging from 1:100 to 1:3200. None of the variables studied were considered to be risk factors for N. caninum. These results show that the prevalence of anti-T. gondii antibodies in swine is high in Paraíba State, that the percentage of viable T. gondii parasites recovered from these animals is also high and that infection by this parasite is closely related to the management of swine. Despite the low prevalence of N. caninum in pigs, further studies are necessary to determine the importance of this parasite for the species.

Coadministration of nematophagous fungi for biological control over gastrointestinal helminths in sheep in the semiarid region of northeastern Brazil.

This study aimed to evaluate coadministration of Duddingtonia flagrans and Monacrosporium thaumasium in a sodium alginate matrix for controlling gastrointestinal helminths in young and adult sheep in the semiarid region of northeastern Brazil. An area of 1ha was divided into two paddocks, in which two experimental groups (fungus and control) were formed, each consisting of six adult females and ten young males. In each group, two subgroups were formed in accordance with the animal category (adult or young). In the fungus group, each animal received 3g of pellets containing 0.6g of fungal mycelium, with 0.3g of D. flagrans and 0.3g of M. thaumasium for each 10 kg of body weight, in their feed twice a week, for six months. In the control group, each animal received 3g of pellets without fungus for each 10 kg of body weight, in their feed twice a week, for six months, serving as a witness group. Reductions in numbers of eggs per gram of feces of 76% among the adult sheep in the fungus group and 83% among the young sheep in the fungus group were observed, in comparison with their respective control subgroups. The groups that received these fungi needed less salvage deworming and presented better packed cell volume percentages, better weight gain and lower levels of L3/kg dry matter in their paddock than the control groups. Thus, it was concluded that coadministration of D. flagrans and M. thaumasium was effective in controlling gastrointestinal helminths of adults and young sheep in the semiarid region of northeastern Brazil.

High genetic diversity in Toxoplasma gondii isolates from pigs at slaughterhouses in Paraíba state, northeastern Brazil: Circulation of new genotypes and Brazilian clonal lineages

The consumption of raw or undercooked pig meat containing Toxoplasma gondii cysts is an important transmission route of this protozoon to animals and humans. This study aimed to serologically diagnose, isolate and genotype T. gondii from pigs slaughtered for human consumption in the state of Paraíba, northeastern Brazil. Blood and tissue samples (heart, tongue and brain) were collected from 120 pigs at slaughterhouses in the state of Paraíba. Serological examinations were performed with an indirect immunofluorescent antibody test (IFAT) with a cut-off point of 1:64. Tissues from positive animals were subjected to bioassays in mice to isolate the parasite. A total of 12.5% (15/120) of the animals were positive according to the IFAT, with titres ranging from 64 to 2048. Viable parasites were isolated in 80% (12/15) of the bioassays. The twelve T. gondii isolates obtained in this study and an additional 13 previously described isolates were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using 11 genetic markers. Additionally, microsatellite (MS) analysis was performed using 15 markers. Nineteen of the 25 isolates completely genotyped using PCR-RFLP had 12 different genotypes, six of which were newly identified. One isolate had a mixed infection. The same 18 non-mixed isolates had 16 different genotypes based on the MS analysis. Genotype #13 (Caribbean 1), which is commonly encountered in northeastern Brazil and is probably a clonal lineage circulating in this region, was the most frequent genotype detected through both the PCR-RFLP and MS analyses. These results demonstrate that T. gondii is widespread among pigs slaughtered in the state of Paraíba. The results also confirm that this parasite has high genetic diversity in this region and that non-archetypal genotypes commonly circulate between different hosts and across different regions of Brazil.

First report of typical Brazilian Toxoplasma gondii genotypes from isolates of free-range chickens ( Gallus gallus domesticus ) circulating in the state of Paraíba, Northeast Brazil

This study evaluated, for the first time, the genetic diversity of Toxoplasma gondii isolates from free-range chickens from the state of Paraíba, Northeast Brazil. Tissue samples from 33 chickens from properties in five municipalities of Paraíba (Esperança, Olho d’Água, Malta, Monteiro, and Patos) were bioassayed in mice. The brains of mice infected with T. gondii cysts were used for DNA extraction and genotyping. Genotyping was performed using 11 PCR-RFLP markers and 15 microsatellite (MS) markers. Complete genotyping results were obtained for 29 isolates, with nine genotypes detected by RFLP and 15 genotypes identified by MS. Three genotypes (#273, #274, and #277) have only been recently identified from pigs in the region. Brazilian clonal types BrII and BrIII were identified from one isolate each. Clonal types I, II, and III were not detected by RFLP. Genotype #13 (Caribbean 1), detected in 48.3% (14/29) of isolates from four of the five municipalities investigated, was the most prevalent genotype in the state of Paraíba. However, the MS analysis showed that of these 14 isolates, only four were unique genotypes, and considering the distance between the municipalities from where they were collected, it is possible that only seven are independent isolates while the others are clones. The other genotypes were restricted to different microregions. The results indicate that the Caribbean 1 lineage of T. gondii is circulating widely in Northeast Brazil. The genotypic diversity of T. gondii in the state of Paraíba is high, and microsatellite analysis revealed this diversity with higher resolution than PCR-RFLP.