Effect of chitosan and Dysphania ambrosioides on the bone regeneration process: A randomized controlled trial in an animal model.

The focus of this triple-blind study was on evaluating the effect of chitosan combined with Dysphania ambrosioides (A) extract on the bone repair process in vivo. In total, 60 male Wistar rats (Rattus norvegicus albinus) weighing between 260 and 270 g were randomly selected for this study and distributed into four groups (n = 15). Group C (chitosan), Group CA5 (chitosan + 5% of D. ambrosioides), Group CA20 (chitosan + 20% of D. ambrosioides), and Group CO (Control-Blood clot). In each animal, bone defects measuring 2 mm in diameter were performed in both tibias for placement of the substances. After 7, 15, and 30 days, the animals were sedated and sacrificed using the cervical dislocation technique and the tissues were analyzed under optical microscope relative to the following events: inflammatory infiltrate, necrosis, osteoclasts, osteoblasts, fibroblasts, periosteal, and endosteal bone formation. The data were evaluated to verify distribution using the Kolmogorov-Smirnov test, and variance, using the Levene test; as distribution was not normal, data were subjected to the Kruskal-Wallis and Dunn nonparametric tests (p < .05). A significant inflammatory infiltrate was observed in Group CA5 (p = .008) in the time interval of 7 days, and in Group C at 15 (p = .009) and 30 (p = .017) days. Osteoblastic activity was more significant in Group CA20 (p = .027) compared with CA5 in the time interval of 7 days. Group CA20 demonstrated a significantly higher endosteal and periosteal bone formation value in the time interval of 7 (p = .013), 15 (p = .004), and 30 days (p = .008) compared with the other groups. The null hypothesis was refuted, bone regeneration was faster in spheres with an association of chitosan and 20% extract, and complete bone repair occurred clinically at 15 days and histologically at 30 days. The spheres proved to be a promising method for the biostimulation of alveolar bone repair and bone fractures.

Histological analysis of biocompatibility of different surgical adhesives in subcutaneous tissue.

The focus of this study was to test the hypothesis that there would be no difference between the biocompatibility of cyanoacrylate-based adhesives in rat subcutaneous tissues. In total, 60 male Wistar rats were used, and divided into four groups (n = 15): Group C (control, PVA-polyvinyl alcohol sponge), Group NO (N-butyl-2-octylcyanoacrylate), Group NH (n-hexyl-cyanoacrylate), and Group EC (Ethyl-cyanoacrylate). The animals were sacrificed after time intervals of 7, 15, and 30 days and tissues were analyzed under optical microscope as regards the events of inflammatory infiltrate, edema, necrosis, granulation tissue, giant cells, young fibroblasts, and collagen formation. The results were statistically analyzed by the Kruskal-Wallis and Dunn tests (p < .05). Significant inflammatory infiltrate was shown for all the adhesives in the time intervals of 7 (p = .004) and 15 days (p = .003). In the time interval of 30 days, moderate inflammatory infiltrate was observed in Groups NH and EC, with significant difference from Control (p = .001). The quantity of collagen fibers in all the experimental groups showed significant difference compared with Control in the time intervals of 7 (p = .002) and 15 days (p = .001), at 30 days only Group EC showed a smaller quantity of collagen fibers in comparison with Control (p = .001). The hypothesis was rejected. The adhesive N-butyl-2-octylcyanoacrylate had less influence on the inflammatory intensity of multinucleated giant cells. Ethyl-cyanoacrylate demonstrated the lowest level of biocompatibility among the adhesives, but its use in clinical practice may be promising for coaptation of smaller edges of superficial tissue. Surgical adhesives were shown to be feasible for clinical use in substitution of conventional suturing. Ethyl-cyanoacrylate should be used with caution due to its greater influence on tissues.

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 study on seroepidemiology and isolation of Toxoplasma gondii in free-range chickens in the semi-arid region of Paraíba state, Brazil.

The aim of this study was to investigate the seroprevalence and isolation of Toxoplasma gondii in free-range chickens in the state of Paraíba, northeastern Brazil. For this, blood samples were collected from 483 chickens in five municipalities in the state of Paraíba. The indirect immunofluorescence assay for anti-T. gondii antibodies was performed. The seropositive birds were slaughtered, and their brains and hearts were collected in order to perform a bioassay in mice. An epidemiological questionnaire was applied on the smallholdings visited, and univariable and multivariable logistic regressions were used to evaluate risk factors. The prevalence of chickens seropositive for T. gondii was found to be 31.5 % (152/483), and 86.1 % (56/65) of the smallholdings were positive. Among the 71 chickens subjected to bioassaying in mice, isolates of T. gondii were obtained from 33 (46.5 %). The isolates were named TgCkBrPB1 to 33. It was observed that the higher the chickens' antibody titer was, the greater the chance of isolating the parasite also was. Sixteen of the 33 isolates (48.5 %) were lethal for all the mice inoculated until 30 days post-inoculation. The risk factors for infection with T. gondii among these free-range chickens were extensive and semi-extensive rearing systems, smallholdings located in urban areas, and presence of cats. The results indicate that the prevalence of T. gondii among chickens in the state of Paraíba is high. Many parasites remained viable in the tissues of the birds studied, and presence of the protozoan was directly related to the management of these birds.