RESUMO O objetivo deste estudo foi isolar Clostridium perfringens e C. difficile em cachorro-do-mato (Cerdocyon thous) da região Nordeste do Brasil. Amostras de fezes de 18 cachorros-do-mato mantidos em cativeiro e oriundos de quatro estados da região Nordeste do Brasil (Alagoas, Bahia, Paraíba e Pernambuco) foram coletadas e submetidas a isolamento de C. perfringens e C. difficile. As colônias sugestivas de C. perfringens foram analisadas para os genes que codificam as principais toxinas de C. perfringens (alfa, beta, épsilon e iota), toxina beta-2 (cpb2), enterotoxina (cpe) e NetB- (netB) e NetF- (netF). As cepas de C. difficile foram analisadas por PCR-multiplex para o gene tpi, toxinas A (tcdA) e B (tcdB) e um gene de toxina binária (cdtB). Alíquotas de amostras de fezes positivas para C. difficile toxigênico foram submetidas a um ELISA comercial para avaliar a presença de toxinas A/B. Clostridium perfringens (tipo A) foi isolado de cinco (27%) amostras, e apenas uma amostra foi positiva para o gene da toxina beta-2 (cpb2). Duas (11%) amostras de fezes foram positivas para C. difficile, mas negativas para toxinas A/B. Estes dois canídeos silvestres também foram positivos para C. perfringens tipo A. Este é o primeiro relato de C. difficile em cachorro-do-mato.

ABSTRACT The aim of the present study was to isolate Clostridium perfringens and C. difficile in crab-eating fox (Cerdocyon thous) from Northeastern Brazil. Stool samples of 18 captive crab-eating foxes from four states of Northeastern Brazil (Alagoas, Bahia, Paraíba e Pernambuco) were collected and subjected to C. perfringens and C. difficile isolation. Suggestive colonies of C. perfringens were then analyzed for genes encoding the major C. perfringens toxins (alpha, beta, epsilon and iota), beta-2 toxin (cpb2), enterotoxin (cpe), and NetB- (netB) and NetF- (netF) encoding genes. C. difficile strains were analyzed by multiplex-PCR for a housekeeping gene (tpi), toxins A (tcdA) and B (tcdB) and a binary toxin gene (cdtB). Unthawed aliquots of stool samples positive for toxigenic C. difficile were subjected to a commercial ELISA to evaluate the presence of A/B toxins. Clostridium perfringens (type A) was isolated from five (27%) samples, and only one sample was positive for beta-2 enconding gene (cpb2). Two (11%) stool samples were positive for C. difficile, but negative for A/B toxins. These two wild canids were also positive for C. perfringens type A. This is the first report of C. difficile in crab-eating fox.

Objetivou-se com este estudo investigar a ocorrência de Mycoplasma spp., Mycoplasma galissepticum (MG) e Mycoplasma synoviae (MS) em psitacídeos de cativeiro localizado no estado de Pernambuco, Brasil. Foram estudadas 85 aves provenientes do Parque Estadual Dois Irmãos, localizado no estado do Pernambuco, Brasil. De cada psitacídeo analisado foram obtidas três amostras por meio de swabs da cloaca, palato e conjuntiva totalizando 255 amostras. As amostras coletadas foram submetidas à extração de DNA e à reação em cadeia da polimerase (PCR), sendo as positivas submetidas ao isolamento em ágar Frey. O DNA de Mycoplasma spp. foi detectado em 16,47% (14/85) dos psitacídeos estudados. Das 255 amostras analisadas, 6,66% (17/255) foram positivas para a presença de Mycoplasma spp., sendo 41,18% (7/17) provenientes da conjuntiva, 35,29% (6/17) do palato e 23,53% (4/17) da cloaca. Nenhuma amostra foi positiva para MG ou MS na PCR. Os resultados obtidos permitem confirmar a presença do DNA de Mycoplasma spp. em conjuntiva, palato e cloaca nas aves estudadas. Foram detectadas colônias semelhantes a membros da classe Mollicutes em 17,64% das amostras (3/17). Esse é o primeiro relato da presença de Mycoplasma spp. em psitacídeos de cativeiro no Nordeste do Brasil.

The aim of this study was to investigate the occurrence of Mycoplasma spp., Mycoplasma galissepticum (MG) and Mycoplasma synoviae (MS) in captive psittacines. Eighty-five wild birds from Parque Estadual Dois Irmãos, Pernambuco state, northeastern Brazil, were used. From each psittacid analyzed three samples were obtained through cloaca, palate and conjunctiva swabs, totaling 255 samples. Samples collected were submitted to DNA extraction and Polimerase Chain Reaction (PCR). Mycoplasma spp. DNA was detected in 16.47% (14/85) of psittacines studied. From 255 samples, 6.66% (17/255) were positive for Mycoplasma spp.: 41.18% (7/17) of positivity in conjunctiva, 35.29% (6/17) in palate and 23.53% (4/17) in cloaca. There was no positive sample for MG or MS in PCR. Similar colonies were found for members of the Mollicutes Class in 17.64% of the samples (3/17). The results confirmed Mycoplasma spp. DNA in conjunctiva, palate and cloaca from the wild birds analyzed. This is the first record of Mycoplasma spp. in captive psittacines from northeastern Brazil.

5-Bromo-2’-deoxyuridine (BrdUrd) has long been known to interfere with cell differentiation. We found that treatment ofBradysia hygida larvae with BrdUrd during DNA puff anlage formation in the polytene chromosomes of the salivary gland S1 region noticeably affects anlage morphology. However, it does not affect subsequent metamorphosis to the adult stage. The chromatin of the chromosomal sites that would normally form DNA puffs remains very compact and DNA puff expansion does not occur with administration of 4 to 8 mM BrdUrd. Injection of BrdUrd at different ages provoked a gradient of compaction of the DNA puff chromatin, leading to the formation of very small to almost normal puffs. By immunodetection, we show that the analogue is preferentially incorporated into the DNA puff anlages. When BrdUrd is injected in a mixture with thymidine, it is not incorporated into the DNA, and normal DNA puffs form. Therefore, incorporation of this analogue into the amplified DNA seems to be the cause of this extreme compaction. Autoradiographic experiments and silver grains counting showed that this treatment decreases the efficiency of RNA synthesis at DNA puff anlages.

RESUMO Beauveria bassiana (Bals.) Vuill. (URM 3447) isolado de Castnia licus (Drury) e reisolado de ovos, larvas e adultos de Anthonomusgrandis (bicudo-do-algodoeiro), artificialmente infectado, foi avaliado sob condições (temperatura: 27 ± 2º C, UR: 60 ± 10% e fotofase: 14h). Estimou-se a viabilidade de B. bassiana (germinação, crescimento colonial e conidiogênese) em meio de cultura Batata-Dextrose-Ágar (BDA) e virulência ao inseto adulto. Conídios em suspensão aquosa com Tween 80 (0,05%), nas concentrações (0,62 x 106; 0,24 x 106 e 0,43 x 106 conídios/mL), constituíram os inóculos para os testes de viabilidade e virulência. Determinou-se a taxa de germinação para o fungo reisolado e não reisolado, 18h após a inoculação. Crescimento das colônias e conidiogênese foram estimados nos intervalos de 3, 6, 9, 12 e 15 dias. As suspensões de conídios nas concentrações aplicadas causaram mortalidade de 96,7, 83,4 e 91,1% em TL de 5,4, 5,9 e 5,3 dias, respectivamente. Verificou-se que B. bassiana (URM-3447), reisolado de ovos, larvas e adultos de A. grandis , apresentou elevada viabilidade de germinação, crescimento colonial e conidiogênese em meio BDA, podendo ser produzido e empregado como agente de controle do inseto.

ABSTRACT Beauveria bassiana (Bals.) Vuill. (URM 3447) isolated from Castnia licus (Drury) and reisolated from eggs, larvae and adults of Anthonomus grandis (boll weevil), infected artificially was evaluated under various conditions (temperature: 27 ± 2º C, HR: 60 ± 10% and photophase: 14h). Estimation was made of the viability of B. bassiana (germination, colonial growth and conidiogenesis) in Potato-Dextrose-Agar (PDA) culture medium and virulence on the adult insect. Conidia in watery suspension with Tween 80 (0.05%), at various concentrations (0.62 x 106; 0.24 x 106 and 0.43 x 106 conidia/mL) constituted the inoculum for the viability and virulence tests. Determination was made of the rate of germination to reisolated and no-reisolated fungus, 18h after the inoculation. Growth of the colonies and conidiogenesis were estimated in the intervals of 3, 6, 9, 12 and 15 days. Suspension of the conidia in concentrations caused mortality of 96.7, 83.4 and 91.1% in LT50 of 5.4, 5.9 and 5.3 days, respectively. The relative pathogen potency (RPP) indicated that the fungus was more aggressive after reisolation. It was verified that B. bassiana (URM-3447), reisolated from eggs, larvae and adults of the A. grandis, presented high viability of germination, colonial growth and conidiogenesis in PDA medium and high degree of virulence on the adult insect, being able to be used as an insect control agent.

O estudo citogenético convencional em uma menina com aniridia esporádica resultou em uma aparente translocação balanceada t(11;13)(p13;q33) de novo. Entretanto, o estudo citogenético pela hibridação in situ fluorescente (FISH) detectou a presença de uma deleção críptica 11p13p14, incluindo a região WAGR e envolvendo aproximadamente 7.5 Mb de DNA, deletando os genes PAX6 e WT1. Estes resultados correlacionam-se com o quadro clínico da paciente e a coloca em alto risco de desenvolver tumor de Wilms. A ausência de retardo mental na paciente indica que a posição distal do ponto de quebra poderá refinar o mapeamento do locus retardo mental na síndrome de genes contíguos WAGR (Wilms, aniridia, anomalias genitais e retardo mental).

Conventional cytogenetic studies on a female infant with sporadic aniridia revealed what appeared to be a balanced de novo t(11;13) (p13;q33) translocation. Fluorescence in situ hybridization (FISH) investigations, however, detected the presence of a cryptic 11p13p14 deletion which included the WAGR region and involved approximately 7.5 Mb of DNA, including the PAX6 and WT1 genes. These results account for the patient's aniridia, and place her at high risk for developing Wilms' tumour. The absence of mental retardation in the patient suggests that the position of the distal breakpoint may also help to refine the mental retardation locus in the WAGR contiguous gene syndrome (Wilms', aniridia, genital anomalies and mental retardation).

When the first group of DNA puffs is active in the salivary gland regions S1 and S3 of Bradysia hygida larvae, there is a large increase in the production and secretion of new salivary proteins demonstrable by [3H]-Leu incorporation. The present study shows that protein separation by SDS-PAGE and detection by fluorography demonstrated that these polypeptides range in molecular mass from about 23 to 100 kDa. Furthermore, these proteins were synthesized mainly in the S1 and S3 salivary gland regions where the DNA puffs C7, C5, C4 and B10 are conspicuous, while in the S2 region protein synthesis was very low. Others have shown that the extent of amplification for DNA sequences that code for mRNA in the DNA puffs C4 and B10 was about 22 and 10 times, respectively. The present data for this group of DNA puffs are consistent with the proposition that gene amplification is necessary to provide some cells with additional gene copies for the production of massive amounts of proteins within a short period of time (Spradling AC and Mahowald AP (1980) Proceedings of the National Academy of Sciences, USA, 77: 1096-1100).

The present paper aimed at testing the action of non-lyophilized venom of Africanized bees Apis mellifera through topical applications on Diatraea saccharalis egg masses. The CL50, DL50 and the most susceptible age of eggs to the venom topic application were also determined. Three-day-old eggs were the most susceptible to the venom action with CL50 equal to 8.6 mg/ml and DL50 equal to 0.173 mg/mass. The venom loses its action after being stored for 15 days.