Molecular and Serologic Detection of Ehrlichia spp. in Endangered Brazilian Wild Captive Felids

Ehrlichiosis, an emergent tickborne disease that affects both humans and animals, may represent a threat to the survival and preservation of wild felids in Brazil There are few studies of ehrlichiosis in wild felids in Brazil, but Ehrlichia spp are present in domestic cats Antibodies to Ehrlichia canis have been reported in a puma (Puma concolor) In this study we assessed the presence of these hemoparasites in the blood of Brazilian wild captive felids of the 72 animals tested, 5 (7%) were seropositive for the E cams antigen, and L1 (15%) were positive for E emirs DNA sequences We also performed sequence alignment to establish the identity of the parasite species infecting these animals using 16S rRNA and omp-1 genes Sequences based on 16S rRNA were similar to those found in dogs and cats from Thailand, Brazil, China, and Taiwan and with E canis obtained from a single individual (human) in Venezuela Ehrlichia sp sequence from sampled felines based on omp-1. gene was similar to the p28 and p30 multigene family of E canis To our knowledge, this is the first study of molecular detection of Ehrlichia sp in Brazilian wild feline species

Molecular characterization of Cryptosporidium spp. in dairy calves from the state of So Paulo, Brazil

Cryptosporidiosis is a common protozoan disease observed in a wide range of vertebrate hosts, including ruminants. Cattle can be a potential reservoir of Cryptosporidium spp., leading to environmental contamination with oocysts of zoonotic species. The molecular characterization of Cryptosporidium spp. isolated from cattle from the state of So Paulo, Brazil, was accomplished using nested polymerase chain reaction for amplification of fragments of the 18S rRNA gene and the glycoprotein GP60 gene, following sequencing of amplified fragments. Positivity for Cryptosporidium was found in 10.7% (21/196) of the samples. Four species of Cryptosporidium were identified: C. andersoni, C. bovis, C. parvum subtype IIaA15G2R1, and C. ryanae. To the best of our knowledge, this is the first report of infection by C. ryanae and C. parvum IIaA15G2R1 in cattle from Brazil.

Potential role for dog fleas in the cycle of Leishmania spp.

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

Structural and ultrastructural characterization of the embryonic development of Pseudoplatystoma spp. hybrids

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

Molecular detection of Cytauxzoon spp. in asymptomatic Brazilian wild captive felids

Cytauxzoon. spp. DNA was detected for the first time in blood samples from asymptomatic Brazilian wild captive felids. In 2606, 72 EDTA blood samples from seven wild felids species: Puma concolor (pinna), Leopardus pardalis (ocelot), Puma yagouaroundi (jaguarundi), Leopardus wiedii (margay), Leopardus tigrinus (little spotted cat), Oncifelis colocolo (pampas cat) and Panthera. onca. (jaguar) were analyzed using polymerase chain reaction to amplify the 18S rRNA gene segment in order to verify the presence of Cytauxzoon spp. DNA. Nine samples were positive: six ocelots, two pumas, and one jaguar. In Brazil, wild felids may be natural reservoirs for Cytauxzoon spp.

Detection of T lymphocytes in intestine of broiler chicks treated with Lactobacillus spp. and challenged with Salmonella enterica serovar enteritidis

The expression of immune response in the form of leukocytic infiltrate by CD3+, CD4+, and CD8+ cells in the epithelium and in the intestinal lamina propria of chicks was studied in the present work by means of immunohistochemical reaction. The chicks were treated with Lactobacillus spp. or cecal microflora (CM) and experimentally challenged or not with Salmonella enterica serovar Enteritidis. The 320 birds utilized were divided into 4 groups containing 80 chicks each and submitted to treatments with Lactobacillus reuteri, Lactobacillus salivarius, Lactobacillus acidophilus, and CM. Each group was subdivided into 4 subgroups of 20 birds each and classified into a subgroup that did not receive treatment (negative control), subgroup treated, subgroup treated and challenged with Salmonella Enteritidis, and subgroup only challenged with Salmonella Enteritidis (positive control). The results obtained show that the treatment with L. reuteri, L. salivarius, L. acidophilus, or CM and challenged or not with Salmonella Enteritidis determine immune response in the form of leukocytic infiltrate by CD3+ and CD8+ lymphocytes followed by CD4+ in the epithelium and in the lamina propria of the duodenum, jejunum, and cecum of chicks up to 12 d of age. The quantity of CD3+ lymphocytes was significantly higher (P < 0.05) in the intestine of chicks treated with L. acidophilus or CM and challenged or not with Salmonella Enteritidis; however, the higher quantity of CD8+ lymphocytes was in the intestine of chicks treated with CM and challenged with Salmonella Enteritidis. The duodenum was the segment in which the immune response by T cells (CD3+, CD4+, and CD8+) was stimulated with the greatest intensity, followed by, respectively, the jejunum and cecum. The quantity of CD3+ lymphocytes present in the duodenum, jejunum, and cecum increases with the age of chicks, independent of the stimulus determined by treatments or challenge.

Molecular detection of Hepatozoon spp. in Brazilian and exotic wild carnivores

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

Antioxidant content in guava (Psidium guajava) and araca (Psidium spp.) germplasm from different Brazilian regions

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Atividade antagônica in vitro de isolados de Trichoderma spp. ao fungo Phytophthora citrophthora

Gummosis is among the main fungal diseases of the citrus. It is caused by Phytophthora sp. and usually shows up in the lap of the plant, provoking rottenness and gum exudation, and expands causing the plant death for constrictions in the cambium or phloem which interrupts the descending flow of sap. The objective of this work was to evaluate the antagonistic in vitro activity of Trichoderma spp. to the fungi Phytophthora citrophthora. Phytophthora citrophthora was exposed to five environments of antagonism (without antagonist and with four strains of Trichoderma viride, T. virens, T. harzianu and T stromaticum), The in vitro essay was accomplished through the method of paired cultures. A completely randomized desing was used with five treatments and three replications, and each plot was represented by three petri dishes. The isolates of Trichoderma demonstrated significant effect in the inhibition of the mycelial growth of the fungi Phytophthora citrophthora, and the fungi Trichoderma stromaticum presented larger antagonism to the fungi P. citrophthora while the T harzianum presented antagonism smaller.

Use of lipopolysaccharides to characterize Bradyrhizobium spp.

Three methods of extraction of lipopolysaccharide (LPS) were compared-the conventional hot phenol-water method with 40% phenol, a modified form of this method using 10% phenol, and the hot saline method. Good recovery of LPS was achieved by each of the three methods, with the LPS found in the aqueous phase with the two phenol-based procedures. The application of SDS-PAGE to the LPS extracts, followed by silver staining, showed similar banding with all three methods of extraction. When the hot saline extraction LPS fraction from eight strains of Bradyrhizobium spp. and eight strains of Bradyrhizobium japonicum was compared with SDS-PAGE, characteristic profiles were achieved. Serological analysis of eight strains of Bradyrhizobium spp., using antisera prepared against whole cells in agglutination reactions, showed extensive sharing of antigens. When antisera was prepared using outer membrane LPS, extracted by the hot saline method, the amount of cross-reaction was reduced greatly. The results indicated that LPS provide an efficient means of obtaining monospecific antisera to be used for serological identification of strains of Bradyrhizobium spp. and that the hot saline extraction method is recommended for a fast, simple and efficient way to obtain LPS and characterize this bacterium.

Wasmannia spp. (Hym., Formicidae) and insect damages to cocoa in Brazilian farms

Diagnostic insect damage to cocoa tree leaves and fruits were compared in situations with and without the presence of the little fire ants, Wasmannia auropunctata and the closely related and sympatric W. cf rochai. No significant differences in thrips, lepidopteran, or chrysomelid beetle damage to fruits, or to young and old leaves, were associated with these ants. However, significant increases of pseudococcids Planococcus citrii, associated with areas dominated by W. auropunctata, and to a lesser degree with W. cf rochai, were present. W. auropunctata has been reported to be a canopy mosaic dominant in cocoa farms, but the lack of reduced herbivore incidences and its lack of spatial permanence do not support favoring its populations for the management of phytophagous insect control.

Prevalence and in vitro antifungal susceptibility of Candida spp isolated from clinical specimens in São Paulo, Brazil

We examined the prevalence and the in vitro susceptibility to antifungal drugs of Candida spp isolated from clinical specimens at our university hospital in São Paulo, Brazil. Among 6,417 samples studied, positive cultures, were obtained from 222 (3.5%) most of them (68%) from the pediatric unit and nursery. Candida albicans and Candida parapsilosis were the most frequent species and the susceptibility patterns of a panel of 130 isolates to amphotericin B, ketoconazole and fluconazole. showed that the order of antifungal efficacy was amphotericin B > ketoconazole > fluconazole.

Padrão de sensibilidade de cepas de Enterobacter spp isoladas na região de Presidente Prudente - SP

Enterobacter spp. are considered important causing agents of infection, specially in hospitalized individuals. The natural resistance of these microorganisms and the great facility to develop resistance to new antimicrobial agents make this genus an important object of study. In this work, 176 strains isolated from various clinical samples were used from hospitalized patients (University Hospital Domingos Leonardo Cerávolo) and from clinic patients (Clinical Laboratory from Unoeste), both situated in Presidente Prudente - SP. E. cloacae (78.9%), E. aerogenes (7.9%) and E. (pantoea) agglomerans (3.9%) were the ones more frequently isolated. Eleven antimicrobial agents were tested by the disk diffusion method and around 90% of the strains presented resistance to the cephalotin, ampicillin and cefaclor. Strains of E. (pantoea) agglomerans presented wide profile of sensibility However one strain of E. cloacae presented resistance to all the antimicrobial agents. The antimicrobial agents with greater inhibitory activity were imipenem and cefepime, for this reason these antimicrobial agents could be the treatment of choice in emergencial therapeutic. This emergencial therapy can be applied with relative security, whereas the data obtained in this study show homogeneity in the profile of sensibility to these antimicrobial agents, independent of the infection site and from the isolated species. The ESBL enzyme could not be detected in no one of the strains by the double diffusion test.

Genetic diversity in Colletotrichum gloeosporioides from Stylosanthes spp. at Centers of origin and utilization

Using molecular markers, this work compares the genetic diversity in Colletotrichum gloeosporioides infecting species of the tropical forage legume Stylosanthes at the center of origin in Brazil and Colombia with that of Australia, China, and India, where Srylosanthes spp. have been introduced for commercial use. There was extensive diversity in the pathogen population from Brazil, Colombia, China, and India. The Australian pathogen population was least diverse probably due to its geographical isolation and effective quarantine. The extensive diversity in China and India means that threats from exotic pathogen races to Stylosanthes pastures can potentially come from countries outside the South American center of origin. In Brazil and India, both with native Stylosanthes populations, a high level of genetic differentiation in the pathogen population was associated with sites where native or naturalized host population was widely distributed. There was limited genetic diversity at germplasm evaluation sites, with a large proportion of isolates having identical haplotypes. This contrasts recent pathogenicity results for 78 of the Brazilian isolates that show hot spots of complex races are more common around research stations where host germplasm are tested, but few are found at sites containing wild host populations. For a pathogen in which the same races arise convergently from different genetic backgrounds, this study highlights the importance of using both virulence and selectively neutral markers to understand pathogen population structure.