Microbiological quality of salmon (Salmo salar) sold in cities of the state of São Paulo, Brazil

The present paper evaluated the microbiology of salmon by quantifying mesophilic heterotrophic microorganisms, total and thermotolerant coliforms, and the presence of Vibrio parahaemolyticus, Staphylococcus aureus, Salmonella sp., Escherichia coli and Aeromonas sp. in the meat. This study can provide technical support for the suggestion of a new regulation of a Brazilian legislation through specific microbiological standards concerning the consumption of raw fish. A number of 31 (16 cooled and 15 frozen) samples of salmon were collected in the retail market network of a few cities in the State of São Paulo, Brazil. Results presented populations of mesophilic heterotrophic microorganisms ranging from 1.0 x 10 and 3.9 x 10(6) CFU/g, total and thermotolerant coliforms in 32.25% and 19.35% of the samples, respectively, and Aeromonas sp. in 41.95% of the samples with a populational variation ranging from 2.0 x 10² to 8.0 x 10³ CFU/g. Staphylococcus aureus was found in one sample whereas Vibrio parahaemolyticus, Salmonella sp. and Escherichia coli were not found. These results demonstrated the presence of potencially pathogenic microorganisms in fresh fish consumed in Brazil, highlighting the necessity of control measures to avoid public health problems related to the consumption of raw fish.

Validação de radioimunoensaio para quantificação de leptina plasmática bovina / Imunoradioassay validation to plasmatic bovine leptin quantification

Devido à necessidade de compreender melhor as interações entre leptina e reprodução, um ria específico para a leptina bovina foi validado. Primeiro, um protocolo para produção de anticorpos foi desenvolvido por meio da inoculação de leptina recombinante equina em um coelho, que resultou em 28,05% de ligação máxima (MB) 105 dias após o inicio do protocolo. Os testes de validação verificaram paralelismo entre a curva padrão e as diluições dos controles alto e baixo (p<0,01). O anticorpo contra leptina equina mostrou especificidade para a leptina bovina (p<0,01). A taxa de recuperação da leptina bovina pelo anticorpo contra leptina recombinante equina foi de 98,4 a 101,6% (p < 0, 01). Quando as amostras foram armazenadas em temperatura ambiente ou refrigeradas à 4°c, foi verificado estabilidade de ligação (p > 0,2), no entanto temperaturas acima de 37°C interferiram negativamente na recuperação da leptina bovina. O uso do tampão de ensaio com ou sem a adição de plasma não apresentou diferenças (p > 0,3). Esses resultados demonstraram que o anticorpo produzido em coelho contra leptina equina foi capaz de detectar a leptina plasmática bovina, e que o ria para a quantificação da leptina bovina apresentou características adequadas para o desenvolvimento de um ensaio válido.

Due necessity of better understanding leptin and reproduction relations, a specific radioimmunoassay (RIA) to bovine leptin was validated. First, an antibody production protocol was developed using recombinant equine leptin inoculated in a rabbit, that results in 28,05% of maximum binding (MB) 105 days after the protocol beginning. The tests of validations verified parallelism between standard curve and dilutions of high and low controls (P < 0,01). Antibody against equine leptin showed specificity to bovine leptin (P < 0,01). The recuperation tax of bovine leptin by antibody against recombinant equine leptin was from 98,4 to 101,6% (P < 0, 01). When the samples were stored in ambient temperature or refrigerated to 4°C, ligation stability was verified (P > 0,2), howether, temperatures above 37°C impaired the bovine leptin recuperation. The use of assay buffer with or without bovine plasma did not show difference (P > 0,3). These results showed that the antibody produced in rabbit against equine leptin were able to detect plasmatic bovine leptin, and that the RIA to bovine leptin quantification had adequate characteristics to the development of a valid assay.

Proteinogramas séricos de ratos Wistar experimentalmente infectados com Trypanosoma evansi / Serum protein concentrations in Wistar rats experimentally infected with Trypanosoma evansi

Estabeleceu-se o perfil eletroforético de proteínas séricas de ratos Wistar experimentalmente infectados com Tripanosoma evansi, utilizando-se 40 ratos, distribuídos em oito grupos de cinco animais cada. Um grupo foi mantido como testemunho (G1), e os demais (G2 a G8) foram inoculados, via intraperitoneal, com cerca de 10³tripomastigota de T. evansi. Amostras de sangue para obtenção de soro foram coletadas no quinto (G2), 10º (G3), 15º (G4), 30º (G5), 45º (G6), 60º (G7) e 75º (G1 e G8) dia após as inoculações. O fracionamento das proteínas foi realizado pela técnica SDS-PAGE. Foram identificadas 31 proteínas, sendo sete de fase aguda: ceruloplasmina (101KD), hemopexina (83KD), transferrina (75KD), albumina (66KD), antitripsina (60KD), haptoglobina (44KD) e glicoproteína ácida (38KD). As proteínas com pesos moleculares 12KD; 22KD; 25KD; 28KD; 32,5KD; 35KD; 53,5KD; 63KD e 72KD apareceram apenas nos ratos inoculados com T. evansi.

This study established the electrophoretic profile of serum proteins of Wistar rats experimentally infected with Tripanosoma evansi. For such, 40 rats were allocated into eight groups of five animals. A group was kept as control (G1) and the others (G2 to G8) were intraperitoneally inoculated with 1.0 x 10³ tripomastigote of T. evansi. Blood samples were collected at 5th (G2), 10th (G3), 15th (G4), 30th (G5), 45th (G6), 60th (G7), and 75th (G1 and G8) days after inoculation (DAI). The serum protein concentrations were determined by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Thirty-one distinct proteins were identified, seven of these were identified as acute phase proteins: ceruloplasmin (110KD), hemopexin (83KD), transferrin (75KD), albumin (66KD), antitrypsin (60KD), haptoglobin (44KD), and acid glycoprotein (38KD). The proteins with molecular weights 12KD; 22KD; 25KD; 28KD; 32,5KD; 35KD; 53,5KD; 63KD, and 72KD were found only in infected rats.

Experiments on intramuscular inoculation and feeding domestic cats (Felis catus) with brains of mice previously infected by rabies viruses

Nineteen kittens divided into four groups were fed with brains of mice infected with rabies viruses. Each four kittens (group I) received four brains infected with the PV fixed strain; nine kittens (group II) ingested 4-5 brains infected with the field isolate T-9/95, isolated from the Desmodus rotundus vampire bat; two kittens (group III) fedten T-9/95-infected brains, and four cats consumed 32-37 PV strain infected brains. One adult male, inoculated into masseter muscle with a 20% T-9/95-infected brain suspension, presented rabies after an incubation period of six days, followed with 8 days of clinical evolution, and died there after and this cat was considered as the rabies “positive standard”. After observing for 20-230 days, all the cats feeding the rabid brains were submitted to euthanasia, by using Acepran®, Zoletil®,and T-61®. At necropsy, samples of brain, heart, lung, kidney, submaxillary salivary gland, and cervical medulla were collected from all the cats and further submitted to the direct fluorescence antibodytest (dFA), mouse inoculation test (MIT) and to the reverse transcriptase-polymerase chain reaction (RT-PCR) technique. Brain, cervical medulla, and the submaxillary salivary gland of the positive standard cat were dFA-positive, and brain and cervical medulla were positive for MIT. All specimens of this cat tested by the RT-PCR were found positive. No animals ingesting PV or T-9/95 virus-infectedbrains developed clinical signs and all materials tested were negativeby dFA and MIT. Several specimens, however, showed positive reactions by the RT-PCR technique, but cats were resistant to rabies through the viruses administered orally.

Dezenove gatos, divididos em quatro grupos, foram alimentadoscom cérebros de camundongos infectados com vírus de raiva. Cada um dos quatro gatos (grupo I) receberam quatro cérebros infectados com vírus fixo PV; nove gatos (grupo II) ingeriram 4-5 cérebros infectados com uma amostra de campo T-9/95, isolada do morcego Desmodus rotundus; dois gatos (grupo III) ingeriram 10 cérebros infectados com T-9/95 e quatro gatos (grupo IV) ingeriram 32-37 cérebros infectados com vírus PV. Um macho adulto, inoculado no músculo masséter, com uma suspensão cerebral a 20% da amostra T-9/95, desenvolveu raiva após período de incubação de seis dias,seguidos por oito dias de evolução clínica, morrendo em seguida. Este gato foi denominado de “padrão positivo”. Após observação por um período de 20-230 dias, todos os gatos que receberam cérebros foram submetidos à eutanásia, utilizando Acepran®, Zoletil® e T-61®. À necropsia, foram colhidas amostras do cérebro, coração, pulmão,rim, glândula salivar submaxilar e medula cervical e submetidas à prova de imunofluorescência direta (IFD), inoculação em camundongos (IC), e reação em cadeia pela polimerase-transcriptase reversa (RT-PCR). No “padrão positivo”, cérebro, medula cervical eglândula salivar foram positivos à IFD e à IC, cérebro e medula cervical foram os positivos. Todos os espécimes do “padrão positivo” foram positivos à RT-PCR. Nenhum animal que ingeriu cérebros contendo amostras de vírus PV ou T-9/95 apresentou sinais clínicos e todos osespécimes testados foram negativos à IFD e IC, no entanto, alguns espécimes reagiram positivamente à RT-PCR, porém, os gatos foram resistentes à raiva com vírus administrados oralmente. Master thesis submitted to the Faculty of Veterinary Medicine and Zootechny of the University of São Paulo, on June 24th, 2003. Fellowship from the Fundação de Amparo à Pesquisa do Estado de São Paulo-FAPESP, process No. 01/07188-8.

Persistence of viable bacilli in highly bacilliferous multibacillary leprosy patients after the twelve doses of multidrug therapy WHO/MDT

The present study evaluate efficacy of twelve doses of multidrug therapy, as suggested by World Health Organization, in ten lepromatous leprosy patients with bacilloscopic index equal or higher than 4. The patients have taken the doses in twelve month-doses regularly. At the end od treatment a specimen was collected by punch biopsy and inoculated into mouse hind footpad acoording to Shepard's technique. Persistence of viable bacilli was demonstrated in three patients. In spite of the small number of patients, same results have been found by other authors, showing that in highly bacilliferous leprosy patients, at the beginning of treatment more doses may be required to prevent relapses.

Persistencia de bacilos viaveis em pacientes de hanseniase multibacilar altamente baciliferos apos doze doses do esquema poliquimioterapico (PQT/OMS) / Persistence of viable bacilli in highly bacilliferous multibacillary leprosy patients after the twelve doses of multidrug therapy WHO/MDT

Trabalho de investigacao, com o obejtivo de avaliar a eficacia do tratamento de doze doses com o esquema poliquimioterapico proposto pela Organizacao Mundial da Saude em 10 pacientes de hanseniase virchoviana em indice baciloscopico igual ou maior que 4. Os pacientes foram submetidos ao tratamento por 12 meses, regularmente. Apos o final do tratamento, foi feita retirada de material e inoculacao em coxim plantar de camundongos, conforme a tecnica de Shepard. Os resultados mostraram persistencia de bacilos viaveis em tres pacientes. Os autores, apesar da amostra pequena, observam que os mesmos resultados tem sido encontrados por outros autores surgerindo que talves seja necessario tratar os pacientes com altos indices baciloscopico no inicio do tratamento, por mais tempo, na tentativa de evitar possiveis recidivas.

Growth and nutritional status of Brazilian wood species Cedrella fissilis and Anadenanthera peregrina in bauxite spoil in response to arbuscular mycorrhizal inoculation and substrate amendment

The growth of Cedrella fissilis Vell. (Cedro Rosa) and of Anadenanthera peregrina Benth (Angico Vermelho) in bauxite spoil was studied to evaluate their response to substrate amendment or to inoculation with arbuscular mycorrhizal fungi (AMF). The plants were grown in bauxite spoil, topsoil or spoil amended with either topsoil or compost, and inoculated with the AMF Acaulospora scrobiculata, Gigaspora margarita or Glomus etunicatum. Root colonization was highly dependent on the interaction plant-fungus-substrate. In C. fissilis, root colonization by Gigaspora margarita dropped from 75(per cent) in bauxite spoil to only 4(per cent) in topsoil. Contrarily, root colonization of A. peregrina by the same fungus increased from 48(per cent) in spoil to 60(per cent) in topsoil. Root colonization of C. fissilis in topsoil was lower than in the three other substrates. The opposite was observed for A. peregrina. Inoculation of the plants with Acaulospora scrobiculata or Glomus etunicatum was very effective in promoting plant growth. Plants of both C. fissilis and A. peregrina did not respond to amendments of bauxite spoil unless they were mycorrhizal. Also, a preferential partitioning of photosynthates to the shoots of A. peregrina inoculated with G. etunicatum or A. scrobiculata, and of C. fissilis inoculated with any of the three species of AMF was observed. C. fissilis showed a greater response to mycorrhizal inoculation than A. peregrina. The mean mycorrhizal efficiency (ME) for dry matter production by C. fissilis was 1,847(per cent) for A. scrobiculata, 1,922(per cent) for G. etunicatum, and 119(per cent) for G. margarita. In A. peregrina, the ME was 249(per cent) for A. scrobiculata, 540(per cent) for G. etunicatum, and 50(per cent) for G. margarita. The effect of mycorrhizal inoculation on plant growth seems to be related in part to an enhanced phosphorus absorption by inoculated plants. Moreover, the efficiency with which the absorbed nutrients were used to produce plant biomass was much greater in plants inoculated with A. scrobiculata or G. etunicatum.