Immobilization of pectinmethylesterase from acerola (Malpighia glabra L.) in porous silica

The total and partially purified enzyme pectinmethylesterase from acerola fruit was covalently immobilized on porous silica particles. These efficiency values were 114% for the total PME and 351% for the partially purified PME. In both forms the immobilization resulted in compounds with high thermal stability.

Prevalence of Sarcocystis neurona and Neospora spp. infection in horses from Brazil based on presence of serum antibodies to parasite surface antigen

Sera from 961 horses from Brazil were tested for antibodies against the major surface antigens SnSAG4 and NhSAG1 to determine the seroprevalence of Sarcocystis neurona and Neospora hughesi, respectively. Antibodies against SnSAG4 were detected in 669 (69.6%) of the horses, while antibodies against NhSAG1 were detected in only 24 (2.5%) of the horses. These serologic results suggest that there is a high concentration of S. neurona in the environment of Brazil, which results in marked exposure of horses to this parasite. Additionally, the data further confirm that infection with Neospora spp. is relatively uncommon in horses. (c) 2005 Elsevier B.V. All tights reserved.

Screening of supports for the immobilization of pectinmethylesterase from acerola (Malpighia glabra L)

A partially purified extract of pectinmethylesterase (PME) from acerola fruit was immobilized on various supports: glass, celite, chrysotile, agarose, concanavalin A Sepharose 4B, egg shell, polyacrylamide and gelatin. In addition, reticulation with glutaraldehyde was assessed, as well as the use of gelatin in the presence of celite, glass and silica. The highest immobilization yields were obtained when the pectinmethylesterase was immobilized in concanavalin A Sepharose 4B (81.7%) and in gelatin-water (78.0%). (C) 2004 Society of Chemical Industry.

Morphological and molecular characterization of Colletotrichum spp. from citrus orchards affected by postbloom fruit drop in Brazil

Brazilian isolates of Colletotrichum spp. from citrus orchards affected by postbloom fruit drop were examined for colony colour, mycelial growth, benomyl-resistance, pathogenicity, and genetic variability by random amplified polymorphic DNA (RAPD) analysis. All isolates were obtained from flowers and persistent calyxes from different citrus hosts from São Paulo, Brazil. DNA polymorphisms detected after amplification with random 10-mer primers were used to classify the isolates into two groups. Group I isolates grew rapidly on potato-dextrose agar (PDA) and were sensitive to benomyl, and group II isolates grew slowly on PDA and were benomyl-resistant. Colletotrichum acutatum was analyzed by RAPD and had high genetic similarity with group II isolates of Colletotrichum from citrus. Probably, the group I is C, gloeosporioides and group II is C. acutatum.

Partial purification and characterization of pectin methylesterase from acerola (Malpighia glabra L.)

The enzyme pectin methylesterase (PME) is present in acerola fruit and was partially purified by gel filtration on Sephadex G-100. The results of gel filtration showed different PME isoforms. The total PME (precipitated by 70% salt saturation) and one of these isoforms (fraction from Sephadex G-100 elution) that showed a molecular mass of 15.5 +/- 1.0 kDa were studied. The optimum pH values of both forms were 9.0. The total and the partially purified PME showed that PME specific activity increases with temperature, the total acerola PME retained 13.5% of its specific activity after 90 min of incubation at 98 degreesC. The partially purified acerola (PME isoform) showed 125.5% of its specific activity after 90 min of incubation at 98 degreesC. The K-m values of the total PME and the partially purified PME isoform were 0.081 and 0.12 mg/mL, respectively. The V-max values of the total PME and the partially purified PME were 2.92 and 6.21 mumol/min/mL/mg of protein, respectively.

Evaluation in vitro of the antagonistic substances produced by Lactobacillus spp. isolated from chickens

To determine the inhibitory capacity of lactic acid bacteria due to the action of antagonistic substances, we tested 474 isolates of Lactobacillus from the crop and cecum of chickens against gram-positive and gram-negative indicator microorganisms by the spot-on-the-lawn and well-diffusion antagonism methods. of the 474 isolates, 265 demonstrated antimicrobial activity against the indicator microorganisms. Isolates identified as L. reuteri, L. salivarius, or Lactobacillus spp. inhibited Enterococcus faecalis, E. faecium, Listeria monocytogenes, and Salmonella spp. but not L. casei, L. delbrueckii, L. fermentum, or L. helveticus by the well-diffusion simultaneous antagonism method under anaerobic incubation conditions. The antagonistic substances produced by some of the Lactobacillus isolates were inactivated after treatment by proteolytic enzymes, which suggested that the substances could be antimicrobial peptides or bacteriocins.

Detection and differentiation of Leptospira spp. serovars in bovine semen by polymerase chain reaction and restriction fragment length polymorphism

In view of the importance of venereal transmission of bovine leptospirosis, the objective of the present study was to apply the polymerase chain reaction (PCR) to 26 serovars of Leptospira interrogans, L. borgpetersenii, L. santarosai, L. noguchii and L. biflexa, to determine the detection threshold in semen samples and to evaluate the possibility of differentiation among serovars using 19 restriction endonucleases. The results showed that all serovars were amplified and the detection threshold in semen samples of a bull was 100 bacteria/ml. Using endonucleases we could classify the 26 serovars into eight groups. The present results show that PCR is a method of great potential for the detection of Leptospira spp, at bovine artificial insemination centers. (C) 2000 Elsevier B.V. B.V. All rights reserved.

Identification and characterization of Colletotrichum spp. affecting fruit after harvest in Brazil

Colletotrichum spp. cause anthracnose in various fruits post-harvest and are a particularly important problem in tropical and subtropical fruits. The disease in fruits of avocado, guava, papaya, mango and passion fruit has been reported to be caused by C. gloeosporioides, and in banana by C. musae. In subtropical and temperate crops such apple, grape, peach and kiwi, the disease is caused by C. acutatum. The variation in pathogenic, morphological, cultural and molecular characteristics of Brazilian isolates of Colletotrichum acutatum Simmonds and isolates from post-harvest decays of avocado, banana, guava, papaya, mango and passion fruit was evaluated. The fruits were inoculated with mycelium of C. acutatum, Colletotrichum spp. and C. musae on a disc of potato dextrose agar. The morphological, cultural and molecular characteristics studied were conidia morphology, colony growth at different temperatures, colony coloration and PCR with primers CaInt2 and ITS4 for C. acutatum and CgInt and ITS4 for C. gloeosporioides. C. acutatum was pathogenic to avocado, guava, papaya, mango and passion fruit, but it was not pathogenic to banana. The morphological, cultural and molecular studies indicated that the avocado, papaya, mango and passion fruit isolates were C. gloeosporioides. The natural guava isolate was identified as C. acutatum, which had not been found previously to produce anthracnose symptoms on guava in Brazil.

The first report of Hepatozoon spp. (Apicomplexa, Hepatozoidae) in domestic cats from São Paulo state, Brazil

Hepatozoon sp. was diagnosed in three naturally infected cats from São Paulo state, Brazil. The first animal was admitted to the veterinary clinic with renal failure. During the hematological examination, gamonts of Hepatozoon sp. were observed within polymorphonuclear cells. Another two cats, which lived in the same house as the first cat, were also positive for this hemoparasite. This is the first report of a Hepatozoon sp. infection in domestic cats from Brazil.

rDNA-based characterization of a new binucleate Rhizoctonia spp. causing root rot on kale in Brazil

In this paper we present the first report of the occurrence of a binucleate Rhizoctonia spp. causing hypocotyl and root rot in kale in Brazil. Rhizoctonia spp. were isolated from kale (Brassica oleracea var. acephala) with symptoms of hypocotyl and root rot. The isolates, characterized as binucleate Rhizoctonia spp., did not show an anastomosis reaction with any of the binucleate Rhizoctonia spp. testers used. The pathogenicity of the isolates was tested under greenhouse conditions; all isolates were pathogenic and showed different symptom severities on kale. The ITS-5.8S rDNA sequences of kale isolates and 50 testers (25 binucleate Rhizoctonia spp. and 25 Rhizoctonia solani) were compared in order to characterize the genetic identity of Rhizoctonia spp. infecting kale. The kale isolates showed genetic identities ranging from 99.3 to 99.8% and were phylogenetically closely related to CAG 7 (AF354084), with identities of 98.5 and 98.7%. It is suggested that the binucleate Rhizoctonia spp. causing hypocotyl and root rot on kale Brazil comprises a new AG not yet described.

Simultaneous Utilization of Galactose and Glucose by Saccharomyces spp.

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)