CARACTERÍSTICAS FÍSICAS E QUÍMICAS DE BEBIDAS LÁCTEAS FERMENTADAS E PREPARADAS COM SORO DE QUEIJO MINAS FRESCAL

Nesta pesquisa procurou-se verificar as características físicas e químicas de bebidas lácteas preparadas com três concentrações de soro de queijo Minas Frescal (30, 40 e 50%), empregando-se dois tipos de culturas lácticas: uma tradicional para iogurte (YC-180) contendo cepas mistas de Streptococcus salivarus subsp. thermophilus, Lactobacillus delbrueckii subsp. lactis e Lactobacillus delbrueckii subsp. bulgaricus e outra (ABY-1) contendo cepas mistas de Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus acidophillus, Bifidobacteria e Streptococcus salivarius subsp. thermophilus. Constatou-se que as bebidas lácteas apresentaram diferença estatística no tempo zero para os teores de gordura e de extrato seco. À medida em que se elevou a proporção de soro em relação ao leite, os teores de gordura e de extrato seco diminuíram. O teor de proteína também diminuiu à medida em que se aumentou o teor de soro nas bebidas lácteas, embora a diferença não tenha sido tão acentuada quanto as observadas para os teores de gordura e de extrato seco. Em relação à lactose, não se constatou diferença entre os tratamentos. Os teores de soro não influenciaram o índice de proteólise das bebidas lácteas. Verificou-se todavia que as bebidas elaboradas com a cultura probiótica ABY-1 apresentaram valores superiores para proteólise quando comparadas às bebidas elaboradas com as culturas YC-180. As bebidas lácteas elaboradas com 30% de soro apresentaram maiores valores para viscosidade. As bebidas elaboradas com a cultura YC-180 apresentaram valores superiores para viscosidade durante o período de armazenamento.

Efeito do teor de sólidos e da concentração de sacarose na acidificação, firmeza e viabilidade de bactérias do iogurte e probióticas em leite fermentado

Doze lotes de leites fermentados foram preparados a 42ºC nos quais as variáveis estudadas foram o teor de sólidos totais (12 e 15%), o teor de sacarose (0% e 8%) e o tipo de co-cultura (Streptococcus thermophilus e Lactobacillus delbrueckii subsp. bulgaricus ; Streptococcus thermophilus e Lactobacillus acidophilus ; Streptococcus thermophilus e Lactobacillus rhamnosus). Parâmetros cinéticos para a diminuição do pH até 4,5 foram calculados. Determinações físico-químicas e microbiológicas foram realizadas após um e sete dias de armazenamento dos produtos a 4ºC. Com o aumento do teor de sólidos totais e adição de sacarose, a atividade de água do leite diminuiu e o tempo para atingir pH 4,5 variou conforme a co-cultura empregada. Os leites fermentados por S. thermophilus e L. acidphilus (STLA) apresentaram pós-acidificação mais acentuada. Aqueles fermentados por S. thermophilus e L. rhamnosus (STLR) foram mais estáveis. Os leites contendo maiores teores de sólidos totais foram aqueles com maior acidez total independente da co-cultura usada. Com o aumento do teor de sacarose e de sólidos solúveis houve um aumento da firmeza usando-se as co-culturas STLR e STLA. Após sete dias, o número de bactérias do iogurte e as probióticas não variou significativamente. Em todos os lotes, o número de bactérias probióticas ficou acima do sugerido pela literatura.

Characterization of goat milk and potentially symbiotic non-fat yogurt

Combining prebiotics and probiotic microorganisms improve quality in the formulation of foods. In this paper, the characteristics of goat milk and symbiotic yogurt were studied. Raw goat milk was analyzed and the skimming process was optimized. For the formulation of a potentially non-fat symbiotic yogurt made with skimmed goat milk, inulin, gelatin, sugar, and Streptococcus salivarius subsp. thermophilus, Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus casei subsp. rhamnoshus. Chemical characteristics, acceptability, and viability of lactic acid bacteria and probiotic culture were assessed. The protein and fat content of the raw milk was 2.90 and 3.56 g/100 mL, respectively. The optimum skimming process was obtained at 9,800 rpm and 4 °C for 15 minutes. The product formulated had a protein and fat content of 4.04 to 0.04 g/100 mL, good sensory properties, and acceptability of 95%. The lactic bacteria count was 9 × 10(7) CFU mL- 1, and probiotic culture count was higher than 1 × 10(6) CFU mL- 1, which guarantees their effect and capacity to survive in the digestive tract and spread in the intestine. The yogurt was stable during the 21 days of storage. Therefore, this study shows that goat milk yogurt is an adequate delivery vehicle of the probiotic culture L. casei and inulin.

"Multiplex PCR" identification of the atypical and monophasic Salmonella enterica subsp. enterica serotype 1,4,[5],12:i:- in São Paulo State, Brazil: frequency and antibiotic resistance patterns

Salmonella spp. are the etiologic agents of salmonellosis, a worldwide spread zoonoses causing foodborne outbreaks and clinical diseases. By serological identification, Salmonella enterica subsp. enterica serotype 1,4,[5],12:i:- accounted for 8.8% of human and 1.6% of nonhuman Salmonella strains isolated in São Paulo State, during 1991-2000. A total of 28.6% of them amplified a fragment corresponding to H:1,2 (flagellar phase two) through PCR analysis and were further assigned as S. Typhimurium. Antimicrobial resistance was detected in 36.3% of the 369 PCR-negative strains tested, including the multiresistance to ampicillin, chloramphenicol, sulfonamides, tetracycline, and streptomycin.

Salmonella enterica subsp houtenae serogroup O:16 in a HIV positive patient: case report

We described a case of salmonellosis in a 33-year old HIV-infected patient. The patient presented oral and esophageal candidiasis, intense epigastric and retrosternal pain. During the physical examination he was hypochloraemic, acyanotic, hypohydrated, anicteric and afebrile. Admittance laboratorial tests indicated: red cells 3.6 millions/mm³; hemoglobin, 10.1 g/dL; leukocyte count, 3,000/mm³, with 1% of eosinophils, 14% of non-segmented and 53% of segmented neutrophils and 31% of lymphocytes. The blood culture was positive for Salmonella enterica subsp houtenae serogroup O:16. This is probably the first human report of bacteremia due to Salmonella enterica subsp houtenae in Brazil associated to HIV-infected patient.

CD19 LYMPHOCYTE PROLIFERATION INDUCED BY Bifidobacterium animalis subsp. lactis IN C57BL/6 MICE EXPERIMENTALLY INFECTED WITH Toxoplasma gondii

Toxoplasmosis is frequently acquired through the oral route by the ingestion of cysts or oocysts of Toxoplasma gondii. Once ingested, the parasites penetrate the intestinal epithelial cells and rapidly disseminate to all organs in the host. During T. gondii infection, the intestinal microbiota plays an important role in stimulating a protective immune response against the parasite. In this sense the use of probiotics is worthy of note since they are live microorganisms that have beneficial effects on the host through stimulation of the immune response that can be important in the control of T. gondii proliferation and dissemination in the host. In the present study, the action of the probiotic Bifidobacterium animalis subsp. lactis was investigated in C57BL/6 mice infected with oocysts of ME49 strain of T. gondii. The probiotic had an immunomodulatory action, inducing CD19 lymphocyte proliferation and consequently increasing anti-T. gondii antibody level.Bifidobacterium animalis subsp. lactisprovided protection in supplemented mice, compared to the control group. In addition, supplemented animals had milder inflammatory process in the small intestine, indicating that the probiotic protects the intestinal mucosa during infection with T. gondii. It was concluded that the probioticB. animalis subsp. lactis induces humoral immune response capable of providing protection against T. gondii infection.

Meiose em Tripsacum australe Cutler e Anderson (T. dactyhides subsp. hispidum Hitchcock)

Depois de uma breve introdução, mostrando a importância que o gênero Tripsacum desempenha hoje nos problemas da origem do milho, fizemos um estudo detalhado da meiose na nova espécie Tripsacum australe, descrita recentemente por CUTLER e ANDERSON (3) e espécie esta encontrada em estado selvagem na América do Sul. Todas as fases da meiose mostraram-se normais e o número cte cromosômios, facilmente determinado nas fases diacinese, metáfase I, metáfase II, é de 18 para a fase haplóide. Esta espécie não difere, quanto ao número de cromosômios, da forma diplóide Tripsacum dactyloide e da espécie Tripsacum floridanum, estudadas por LONGLEY (5). Segundo MANGELSDORF e REEVES (10) as formas de Tripsacum encontradas na América Central têm 72 cromosômios e são consideradas como autotetraplóides. Entretanto, no que se refere à presença de "knobs" nos cromosômios, esta espécie parece diferir da espécie estudada por LONGLEY (5). Tripsacum australe não apresenta "knobs" nas extremidades dos cromosômios e provavelmente também nas outras regiões pois as figuras que puderam ser examinadas não mostraram essa estrutura. Segundo MANGELSDORF e REEVES (10) os "knobs" presentes no milho teriam vindo de Tripsacum, por meio de cruzamento entre estes dois gêneros. Assim sendo, os tipos de milho cultivados próximos ao centro de distribuição das espécies de Tripsacum até então conhecidas, e que é a região da América Central, principalmente o México, deveriam se apresentar bastante contaminados por este gênero e apresentariam mais "knobs" do que aqueles tipos de milho cultivados ionge da referida região. Observações de vários autores (6, 7, 9, 20, 11 e 12) confirmam esta hipótese, inclusive aquelas realizadas por um dos autores deste trabalho (Graner, não publicado) em material sul-americano. Tendo sido encontrada agora esta nova espécie de Tripsacum na América do Sul, aparentemente sem "knobs", torna-se interessante verificar se ela não poderia ter contribuido para o estabelecimento das formas de milho sem "knobs" encontradas na América do Sul. Cruzamentos entre milho e Tripsacum australe foram realizados por um dos autores (Addison), não lendo porém produzido sementes. Outras pesuisas tornam-se então necessárias afim de que se possa tirar conclusões a respeito de tão importante assunto.

Biochemical, immunological and toxicological characteristics of the crystal proteins of Bacillus thuringiensis subsp. medellin

Characterization of the insecticidal and hemolytic activity of solubilized crystal proteins of Bacillus thuringiensis (Bt) subsp. medellin (Btmed) was performed and compared to solubilized crystal proteins of isolates 1884 of B. thuringiensis subsp. israelensis (Bti) and isolate PG-14 of B. thuringiensis subsp. morrisoni (Btm). In general, at acid pH values solubilization of the Bt crystalline parasporal inclusions (CPI) was lower than at alkaline pH. The larvicidal activity demonstrated by the CPI of Btmed indicated that optimal solubilization of CPI takes place at a pH value of 11.3, in Bti at pH values from 5.03 to 11.3 and in Btm at pH values from 9.05 to 11.3. Hemolytic activity against sheep red blood cells was mainly found following extraction at pH 11.3 in all Bt strains tested. Polyacrylamide gel electrophoresis under denaturing conditions revealed that optimal solubilization of the CPI in all Bt strains takes place at the alkaline pH values from 9.05 to 11.3. An enriched preparation of Btmed crystals was obtained, solubilized and crystal proteins were separated on a size exclusion column (Sephacryl S-200). Three main protein peaks were observed on the chromatogram. The first peak had two main proteins that migrate between 90 to 100 kDa. These proteins are apparently not common to other Bt strains isolated to date. The second and third peaks obtained from the size exclusion column yielded polypeptides of 68 and 28-30 kDa, respectively. Each peak independently, showed toxicity against 1st instar Culex quinquefasciatus larvae. Interestingly, combinations of the fractions corresponding to the 68 and 30 kDa protein showed an increased toxicity. These results suggest that the 94 kDa protein is an important component of the Btmed toxins with the highest potency to kill mosquito larvae. When crystal proteins of Bti were probed with antisera raised independently against the three main protein fractions of Btmed, the only crystal protein that showed cross reaction was the 28 kDa protein. These data suggest that Btmed could be an alternative bacterium for mosquito control programs in case mosquito larval resistance emerges to Bti toxic proteins.

Cloning, Expression and Toxicity of a Mosquitocidal Toxin Gene of Bacillus thuringiensis subsp. medellin

Bacillus thuringiensis (Bt) subsp. medellin (Btmed) produces parasporal crystalline inclusions which are toxic to mosquito larvae. It has been shown that the inclusions of this bacterium contain mainly proteins of 94, 68 and 28-30 kDa. EcoRI partially digested total DNA of Btmed was cloned by using the Lambda Zap II cloning kit. Recombinant plaques were screened with a mouse policlonal antibody raised against the 94 kDa crystal protein of Btmed. One of the positive plaques was selected, and by in vivo excision, a recombinant pBluescript SK(-) was obtained. The gene encoding the 94 kDa toxin of Btmed DNA was cloned in a 4.4 kb DNA fragment. Btmed DNA was then subcloned as a EcoRI/EcoRI fragment into the shuttle vector pBU4 producing the recombinant plasmid pBTM3 and used to transform by electroporation Bt subsp. israelensis (Bti) crystal negative strain 4Q2-81. Toxicity to mosquito larvae was estimated by using first instar laboratory reared Aedes aegypti, and Culex quinquefasciatus larvae challenged with whole crystals. Toxicity results indicate that the purified inclusions from the recombinant Bti strain were toxic to all mosquito species tested, although the toxicity was not as high as the one produced by the crystal of the Btmed wild type strain. Poliacrylamide gel electrophoresis indicate that the inclusions produced by the recombinant strain Bti (pBTM3) were mainly composed of the 94 kDa protein of Btmed, as it was determined by Western blot

Antimicrobial susceptibility of Campylobacter jejuni subsp. jejuni assessed by E-test and double dilution agar method in Southern Chile

The susceptibility patterns of 108 Campylobacter jejuni subsp. jejuni clinical strains, to six antimicrobial agents was determined by using the E-test and the double dilution agar methods. Using both metods, no strain was found to be resistant to ciprofloxacin, erythromycin and gentamicin, but two (1.8%) were resistant to tetracycline and all to aztreonam. Seven (6.5%) strains were resistant to ampicillin by the E-test and five (4.6%) by the double dilution agar method and by both meyhods. No great discrepancies were observed between both methods.

Proteolytic processing of the Cyt1Ab1 toxin produced by Bacillus thuringiensis subsp. medellin

Bacillus thuringiensis produces d-endotoxins that require proteolytic processing to become active. The activation of the B. thuringiensis subsp. medellin 28 kDa (Cyt1Ab1) cytolytic toxin by trypsin, chymotrypsin and gut extract from Culex quinquefasciatus larvae was analyzed. The Cyt1Ab1 toxin of B. thuringiensis subsp. medellin was processed by all proteases tested to fragments between 23 and 25 kDa, while processing of the Cyt1Aa1 toxin produce fragments between 22.5 and 24.5 kDa. The Cyt1Ab1 toxin was preferentially processed at the alkaline pH of 12. The in vitro proteolytic processing of the Cyt1Ab1 toxin by C. quinquefasciatus larvae midgut extract showed a 25 kDa fragment; a similar result was observed when the activation was performed in the in vivo experiments. The solubilized Cyt1Ab1 toxin and the protease resistant cores generated by in vitro processing showed hemolytic activity but not mosquitocidal activity. Amino terminal sequence of the C. quinquefasciatus gut extract resistant fragment indicated that the cutting site was located between Lys31 and Asp32, with a sequence DDPNEKNNHNS; while for the trypsin-resistant fragment the cutting site was determined between Leu29 and Arg30, and for the chymotrypsin-resistant fragment between Arg30 and Lys31.

A theoretical model of the tridimensional structure of Bacillus thuringiensis subsp. medellin Cry 11Bb toxin deduced by homology modelling

Cry11Bb is an insecticidal crystal protein produced by Bacillus thuringiensis subsp. medellin during its stationary phase; this ¶-endotoxin is active against dipteran insects and has great potential for mosquito borne disease control. Here, we report the first theoretical model of the tridimensional structure of a Cry11 toxin. The tridimensional structure of the Cry11Bb toxin was obtained by homology modelling on the structures of the Cry1Aa and Cry3Aa toxins. In this work we give a brief description of our model and hypothesize the residues of the Cry11Bb toxin that could be important in receptor recognition and pore formation. This model will serve as a starting point for the design of mutagenesis experiments aimed to the improvement of toxicity, and to provide a new tool for the elucidation of the mechanism of action of these mosquitocidal proteins.

In vivo binding of the Cry11Bb toxin of Bacillus thuringiensis subsp. medellin to the midgut of mosquito larvae (Diptera: Culicidae)

Bacillus thuringiensis subsp. medellin produces numerous proteins among which 94 kDa known as Cry11Bb, has mosquitocidal activity. The mode of action of the Cry11 proteins has been described as similar to those of the Cry1 toxins, nevertheless, the mechanism of action is still not clear. In this study we investigated the in vivo binding of the Cry11Bb toxin to the midgut of the insect species Anopheles albimanus, Aedes aegypti, and Culex quinquefasciatus by immunohistochemical analysis. Spodoptera frugiperda was included as negative control. The Cry11Bb protein was detected on the apical microvilli of the midgut epithelial cells, mostly on the posterior midgut and gastric caeca of the three mosquito species. Additionally, the toxin was detected in the Malpighian tubules of An. albimanus, Ae. aegypti, Cx. quinquefasciatus, and in the basal membrane of the epithelial cells of Ae. aegypti midgut. No toxin accumulation was observed in the peritrophic membrane of any of the mosquito species studied. These results confirm that the primary site of action of the Cry11 toxins is the apical membrane of the midgut epithelial cells of mosquito larvae.