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.

Novos Registros sobre Meloidogyne mayaguensis no Brasil e Estudo Morfológico Comparativo com M. incognita

Almeida, E.J., P.L.M. Soares, A.R. Silva & J.M. Santos. 2008. New records on Meloidogyne mayaguensis in Brazil and comparative study with M. incognita. Meloidogyne mayaguensis is the plant-parasitic nematode responsible for a great impact on guava production in Brazil, since it has been the cause for eradication of thousand of hectares of guava plantations in the Northeastern region. In the present study, this species was detected in soybean fields of Ituverava municipality, São Paulo state, and in different vegetable crops (lettuce, cucumber, pepper and cherry tomato) in Chapada dos Guimaraes municipality, Mato Grosso state, causing root galls and other symptoms. The species was identified on the basis of the perineal pattern of females, and on the morphology and morphometry of anterior region of males. Isozyme phenotype for esterase was used for confirmation. This constitutes the first report on the occurrence of M. mayaguensis on lettuce, cucumber, pepper and cherry tomato cultures in Mato Grosso state and the first one in soybean in São Paulo state. It was found that morphological features of male anterior region and female perineal pattern are enough for the safe distinction between M. mayaguensis and M. incognita.

Avaliação Comparativa da Agressividade de Meloidogyne javanica e M. incognita à Variedade SP 911049 de Cana-de-açúcar

Nematodes cause extensive losses to sugarcane in Brazil and also in other producing regions. Meloidogyne incognita, M. javanica and Pratylenchus zeae are the key species for this culture worldwide. In the present study, the aggressiveness of M. javanica and M. incognita to sugarcane variety SP 911049 was evaluated comparatively,. The following parameters were compared: reproduction factor (RF) of these nematodes, effect of nematodes in the natural incidence of pests, and the influence on the development and technological characteristics of sugarcane. Considering the data of RF, biometrics, natural infestation of pests, mortality of plants, and technological variables, it was concluded that M. javanica was more aggressive to sugarcane, although its rate of multiplication was much smaller than the one of M. incognita.

Flutuação populacional de Meloidogyne enterolobii em pomar de goiabeira (Psidium guajava)

The objective of this work was to study the population fluctuation of Meloidogyne enterolobii in a commercial orchard of 'Paluma' guava with five years of age, under micro sprinkler irrigation system and usual practices for this crop. In the orchard, located in the municipality of Vista Alegre do Alto (SP) Brazil and infested by M. enterolobii, five plants exhibiting nematode symptoms were selected for monthly sampling from July 2007 to June 2008. Two single samples of rhizosphere soil and guava roots were collected from each plant, with the aid of hoe to a depth of 20 cm on the projection of the trees, forming a composite sample per plant. In nematology laboratory of Faculdade de Ciencias Agrarias e Veterinarias Universidade Estadual Paulista, eggs and second stage juveniles (J2) of M. enterolobii were extracted from the samples and quantified. There was not a marked seasonal variation in the nematode population in soil or roots, possibly as a result of the guava vegetation stimulated by irrigation or the presence of invasive plants during the experimental period. However, the soil density reached a peak in January, corresponding to the month of elevated rainfall and temperatures. In roots, there were two significant peaks, in February and a major peak in August. There was a negative correlation between nematode populations in soil and roots.

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.