Cinética ruminal de dietas contendo farelo de mamona destoxificado / Ruminal kinetics of diets containing detoxicated castor bean meal

The kinetics of gas production and in vitro ruminal degradation of diets contained replacement of soybean meal (SBM) by detoxicated castorbean meal (DCBM) (0, 15, 30 and 45% of substitution) were evaluated in this study. Values of gas production of non-fibrous carbohydrates (Vf1), fibrous carbohydrates (Vf2) and total carbohydrates (V) showed quadratic behavior, and by observation of equations can be obtained inclusions of 25% the V and Vf1 obtained maximum values of 174.7 and 237.8 ml gas / g DM, respectively. The potential degradability (PD) and effective degradability (ED) of dry matter showed quadratic behavior and by observation of equations can be perceive that up to 18.0% there was an increase in the values of PD and ED that obtained maximum values of 93.7% and 55.7%, respectively. The replacement of SBM by DCBM from 15 to 30% increased the kinetics of gas production and rumen degradation.(AU)

Isolation, enumeration, molecular identification and probiotic potential evaluation of lactic acid bacteria isolated from sheep milk

Lactic acid bacteria species were molecularly identified in milk from Lacaune, Santa Inês and crossbred sheep breeds and their in vitro probiotic potential was evaluated. The species identified were Enterococcus faecium (56.25%), E. durans (31.25%) and E. casseliflavus (12.5%). No other lactic acid bacteria species, such as lactobacilli, was identified. Most of the isolated enterococci were resistant to gastric pH (2.0) and to 0.3% oxgall. All tested enterococci were resistant to ceftazidime, oxacillin and streptomycin and sensible to clindamycin, erythromycin and penicillin. The resistance to ciprofloxacin, gentamicin, tetracycline and vancomycin varied among tested species. All tested enterococci strongly inhibited (P<0.05) Escherichia coli and Listeria monocytogenes, moderately inhibited E. faecalis and Staphylococcus aureus and did not inhibit Pseudomonas aeruginosa, Salmonella enterica var. Typhimurium and also one E. durans sample isolated from sheep milk. Four samples of E. faecium, one of E. durans and one of E. casseliflavus presented the best probiotic potential...

Espécies de bactérias ácido-lácticas foram identificadas em nível molecular em leite das raças ovinas Lacaune, Santa Inês e suas mestiças, e o seu potencial probiótico in vitro foi avaliado. As espécies identificadas foram Enterococcus faecium (56,25%), E. durans (31,25%) e E. casseliflavus (12,5%). Nenhuma outra espécie de bactéria ácido-láctica, como Lactobacillus sp., foi identificada. A maioria dos enterococos isolados foi resistente ao pH gástrico (2.0) e a 0,3% de oxgall. Todos os enterococos testados foram resistentes à ceftazidima, oxacilina e estreptomicina e sensíveis à clindamicina, eritromicina e penicilina. A resistência à ciprofloxacina, gentamicina, tetraciclina e vancomicina variou entre as amostras. Todos os enterococos testados inibiram fortemente (P<0,05) Escherichia coli e Listeria monocytogenes, inibiram moderadamente E. faecalis e Staphylococcus aureus e não inibiram Pseudomonas aeruginosa, Salmonella enterica var. Typhimurium e uma amostra de E. durans isolada de leite de ovelha. Quatro amostras de E. faecium, uma de E. durans e uma de E. casseliflavus apresentaram o melhor potencial probiótico...

Probiotic technological and functional characteristics of Lactobacillus strains isolated from chicken gut / Características tecnológicas e funcionais de linhagens de Lactobacillus isoladas do trato gastrointestinal de frangos

Lactobacillus spp. isolated from different portions of chickens' gastrointestinal tract were evaluated concerning their ability to survive in a water-in-oil (W/0) emulsion containing sesame and sunflower oil. After sixty days of emulsion storage under refrigeration, three of five strains tested survived in number equal to or higher than 10(6)cfu/g. Lactobacillus reuteri 2M14C, which presented the highest survival in W/O emulsion (10(7)cfu/g), was tested for its capacity to resist throughout the passage through gnotobiotic mice gastrointestinal tract and for the ability to stimulate murine peritoneal macrophages phagocytosis. This strain remained at a number above 10(9)cfu/g feces during ten days of monoassociation, and monoassociated mice showed phagocytic activity significantly greater than the germ-free controls (P<0.05). The results suggest that the formulation can be used to incorporate viable Lactobacillus spp. cells in animal feed. Moreover, the results suggest that L. reuteri 2M14C is a strong candidate to be incorporated in probiotic formulations for use in chicken.

Lactobacillus spp. isolados de diferentes porções do trato gastrintestinal de frangos foram testados quanto à capacidade de se manterem viáveis em uma emulsão água/óleo (A/O) contendo óleos de gergelim e de girassol. Após sessenta dias de estocagem sob refrigeração, três de cinco linhagens testadas sobreviveram em concentração igual ou superior a 10 6 UFC/g. Lactobacillus reuteri 2M14C, que apresentou maior capacidade de sobrevivência na emulsão desenvolvida (10 7 UFC/g), foi testado quanto à sua capacidade de sobreviver às condições do trato gastrintestinal in vivo em camundongos gnotobióticos. Após dez dias de monoassociação com L. reuteri 2M14C, foi testada também a capacidade de estimulação da atividade fagocítica de macrófagos peritoneais. A linhagem permaneceu em número superior a 10 9 UFC/g de conteúdo fecal durante os dez dias de monoassociação, e os camundongos monoassociados apresentaram atividade fagocítica maior (P<0,05) que a do grupo controle isento de germe. Os resultados sugerem que a formulação proposta é capaz de manter a viabilidade de células de lactobacilos para adição em ração animal, necessitando, no entanto, de um acompanhamento dessa viabilidade por tempo maior de estocagem. Além disso, os resultados demonstram que Lactobacillus reuteri 2M14C é um forte candidato a ser adicionado em formulações probióticas para uso em frangos.

Potencial probiótico in vitro de bactérias ácido-láticas isoladas de queijo-de-minas artesanal da Serra da Canastra, MG / Probiotic potential of lactic acid bacteria isolated from minas artisanal cheese from Serra da Canastra, MG

O potencial probiótico in vitro de 12 amostras de bactérias ácido-láticas (11 Lactobacillus spp. e uma Weissella paramesenteroides), isoladas de queijo-de-minas artesanal da Serra da Canastra, foi investigado. Essas amostras foram caracterizadas in vitro quanto à susceptibilidade a antimicrobianos, antagonismo contra microrganismos de referência patogênicos e não patogênicos e sensibilidade a pH gástrico e sais biliares. Apenas L. rhamnosus B4, W. paramesenteroides C10 e L. rhamnosus D1 apresentaram resistência a menor número de antimicrobianos (ceftazidima, oxacilina, estreptomicina e vancomicina). Todas as amostras apresentaram atividade antagonista frente a todas as bactérias patogênicas testadas e não inibiram as bactérias não patogênicas. Todas as amostras foram resistentes ao pH gástrico (2.0). Alguns microrganismos mostraram pouca inibição do crescimento em presença de sais biliares, enquanto outros foram moderadamente ou altamente inibidos. L. rhamnosus B4, W. paramesenteroides C10 e L.rhamnosus D1 apresentaram o melhor potencial probiótico de acordo com os testes in vitro realizados.

The in vitro probiotic potential of twelve samples of lactic acid bacteria (eleven Lactobacillus spp. and one Weissella paramesenteroides) isolated from Minas artisanal cheese from Serra da Canastra was evaluated for their antimicrobial susceptibility, antagonistic activity against pathogenic and lactic acid bacteria, as well as for their tolerance to gastric pH and biliary salts. Only L. rhamnosus B4, W. paramesenteroides C10, and L. rhamnosus D1 showed resistance to tested antimicrobials (ceftazidime, oxacillin, streptomycin, and vancomycin). All samples showed antagonism against all pathogenic bacteria tested and did not inhibit the growth of nonpathogenic samples. Regarding gastric pH tolerance, all samples showed low inhibition at pH 2. The tolerance to biliary salts varied from low to great among tested samples. L. rhamnosus B4, W. paramesenteroides C10, and L. rhamnosus D1 showed the best probiotic potential revealed by their in vitro test results.

Anemia mielotísica por osteopetrose em um cão: relato de caso / Mielotisic anemia caused by osteopetrosis in a dog: case report

A osteopetrose é uma doença rara, caracterizada pelo aumento generalizado da densidade óssea. Tem como característica principal, a reabsorção osteoclástica defeituosa, resultando no acúmulo de massa óssea. Além disso, pode ocorrer retardo do crescimento, desnutrição progressiva, anemia e caquexia. O presente relato descreve o caso de uma cadela, com aproximadamente nove meses de idade, sem raça definida, com histórico de apatia e disorexia. Hemogramas seriados demonstraram pancitopenia persistente. Vários exames laboratoriais foram realizados para excluir doenças como erliquiose, leishmaniose e cinomose, porém todos foram negativos. O mielograma constatou hipocelularidade relativa por provável aplasia/hipoplasia medular. Exames radiográficos evidenciaram o aumento da radiopacidade óssea e hipertrofia da região cortical, e a necropsia confirmou a redução do canal medular. O exame histopatológico confirmou a osteopetrose. Conclui-se que a osteopetrose pode causar comprometimento na produção de células sanguíneas, sendo uma causa rara de anemia mielotísica, além de alterações neurológicas secundárias à má-formação dos ossos do crânio.

Osteopetrosis is a rare disease characterized by generalized increase in bone density. The defective osteoclastic resorption results in the accumulation of bone mass. Furthermore, there may be growth delay, progressive malnutrition, anemia and cachexy. This report describes the case of a nine month old, mixed breed, female dog presented with apathy and disorexia. Laboratory tests were performed to rule out Ehrlichiosis, Leishmaniasis, and Canine distemper, but all were negative. Hemogram showed persistent pancytopenia and myelogram showed relative hypocellularity, probably due to spinal cord aplasia/hypoplasia. Radiographs showed increased bone opacity and hypertrophy of the cortical region, and the necropsy confirmed the reduction of the medullary canal. Histopathological examination confirmed osteopetrosis. In conclusion, osteopetrosis can compromise blood cell production, and is a rare cause of mielotisic anemia. It can also cause neurological deficits due to cranium bone deformation.

Molecular identification of Lactobacillus spp. associated with puba, a Brazilian fermented cassava food

Puba or carimã is a Brazilian staple food obtained by spontaneous submerged fermentation of cassava roots. A total of 116 lactobacilli and three cocci isolates from 20 commercial puba samples were recovered on de Man, Rogosa and Sharpe agar (MRS); they were characterized for their antagonistic activity against foodborne pathogens and identified taxonomically by classical and molecular methods. In all samples, lactic acid bacteria were recovered as the dominant microbiota (7.86 ± 0.41 log10 CFU/g). 16S-23S rRNA ARDRA pattern assigned 116 isolates to the Lactobacillus genus, represented by the species Lactobacillus fermentum (59 isolates), Lactobacillus delbrueckii (18 isolates), Lactobacillus casei (9 isolates), Lactobacillus reuteri (6 isolates), Lactobacillus brevis (3 isolates), Lactobacillus gasseri (2 isolates), Lactobacillus nagelii (1 isolate), and Lactobacillus plantarum group (18 isolates). recA gene-multiplex PCR analysis revealed that L. plantarum group isolates belonged to Lactobacillus plantarum (15 isolates) and Lactobacillus paraplantarum (3 isolates). Genomic diversity was investigated by molecular typing with rep (repetitive sequence)-based PCR using the primer ERIC2 (enterobacterial repetitive intergenic consensus). The Lactobacillus isolates exhibited genetic heterogeneity and species-specific fingerprint patterns. All the isolates showed antagonistic activity against the foodborne pathogenic bacteria tested. This antibacterial effect was attributed to acid production, except in the cases of three isolates that apparently produced bacteriocin-like inhibitory substances. This study provides the first insight into the genetic diversity of Lactobacillus spp. of puba.

Queijo de minas artesanal da Serra da Canastra: influência da altitude das queijarias nas populações de bactérias acidolácticas / Influence of altitute on lactic acid bacteria population of minas artisanal cheese from Serra da Canastra

Amostras de queijo de minas artesanal foram coletadas em 18 queijarias localizadas em propriedades rurais da região da Serra da Canastra, Minas Gerais, com o objetivo de avaliar a influência da altitude sobre a população de bactérias acidolácticas. As queijarias estavam distribuídas nas altitudes de 600 a 900m, 900 a 1000m e mais de 1000m. Observaram-se populações mais elevadas de bactérias acidolácticas nas amostras de queijo da altitude de 600 a 900m. Lactobacillus rhamnosus, Lactobacillus casei e Lactobacillus plantarum foram os principais microrganismos isolados e identificados por PCR ARDRA 16S-23S rDNA, além de Enterococcus spp., Lactococcus spp. e outras espécies de Lactobacillus. Sugere-se que estas espécies estejam adaptadas ao ambiente de produção do queijo de minas artesanal produzido na região, o que resultaria em características sensoriais próprias do produto.

Samples of minas artisanal cheese were collected in 18 small-scale producer properties located in the rural region of Serra da Canastra, Minas Gerais state, aiming to evaluate the influence of three altitudes, from 600 to 900m, 900 to 1000m, and higher than 1000m, on the lactic acid bacteria (LAB) population. High populations of LAB were observed in the cheese samples, mainly in the lowest altitude. Lactobacillus rhamnosus, Lactobacillus casei, and Lactobacillus plantarum were the major LAB isolated from the cheese samples and identified according to PCR ARDRA 16S-23S rDNA. Enterococcus spp., Lactococcus spp., and other species of Lactobacillus genus were also found. It is suggested that these microorganisms are adapted to the production environment of the minas artisanal cheese which result in the unique sensorial properties of the product.

Identification and in vitro production of Lactobacillus antagonists from women with or without bacterial vaginosis

Lactobacilli isolated from the vaginal tract of women with and without bacterial vaginosis (BV) were identified and characterized for the production of antagonists. Bacterial samples were isolated from healthy women (N = 16), from patients with clinical complaints but without BV (N = 30), and from patients with BV (N = 32). Identification was performed using amplified ribosomal DNA restriction analysis. Production of antagonistic compounds was evaluated by the double-layer diffusion technique using Gram-positive (N = 9) and Gram-negative bacteria (N = 6) as well as yeast (N = 5) as indicator strains. Of a total of 147 isolates, 133 were identified as pertaining to the genus Lactobacillus. Lactobacillus crispatus was the species most frequently recovered, followed by L. johnsonii and L. jensenii. Statistical analysis showed that L. crispatus was more frequent in individuals without BV (P < 0.05). A higher production of antagonists was noted in L. crispatus isolates from healthy women (P < 0.05). More acidic local pH and higher H2O2 production by isolated lactobacilli from healthy women suggest these mechanisms as the possible cause of this antagonism. In conclusion, a significant correlation was detected between the presence and antagonistic properties of certain species of Lactobacillus and the clinical status of the patients.

Atividade antimicrobiana de bactérias ácido-lácticas isoladas de queijos de coalho artesanal e industrial frente a microrganismos indicadores / Antimicrobial activity of lactic acid bacteria isolated from artisanal and industrial

Quatro cepas de Lactobacillus spp. e duas cepas de Lactococcus spp. isoladas de queijos de coalho artesanal e industrial foram testadas quanto às suas atividades antimicrobianas. Observou-se atividade antagonista dessas bactérias ácido-lácticas frente a elas, a outras bactérias ácido-lácticas isoladas de queijo de coalho, aos patógenos isolados dos mesmos queijos e a cepas de patógenos de referência. Verificou-se diferença (P<0,05) entre as atividades antagonistas, exceto quando as próprias bactérias ácido-lácticas foram utilizadas como reveladoras. Lactobacillus spp. apresentaram as atividades antagonistas mais potentes.

The association of genetic markers and malaria infection in the Brazilian Western Amazonian region

Almost all individuals (182) belonging to an Amazonian riverine population (Portuchuelo, RO, Brazil) were investigated for ascertaining data on epidemiological aspects of malaria. Thirteen genetic blood polymorphisms were investigated (ABO, MNSs, Rh, Kell, and Duffy systems, haptoglobins, hemoglobins, and the enzymes glucose-6-phosphate dehydrogenase, glyoxalase, phosphoglucomutase, carbonic anhydrase, red cell acid phosphatase, and esterase D). The results indicated that the Duffy system is associated with susceptibility to malaria, as observed in other endemic areas. Moreover, suggestions also arose indicating that the EsD and Rh loci may be significantly associated with resistance to malaria. If statistical type II errors and sample stratification could be ruled out, hypotheses on the existence of a causal mechanism or an unknown closely linked locus involved in susceptibility to malaria infection may explain the present findings

Complement resistance of Leishmania amazonensis promastigotes is independent of parasite proteases and lysis of sensitive forms is not due to natural antibodies in normal human serum

Leishmania amazonensis promastigotes cultivated in vitro differentiate from complement-sensitive to complement-resistant forms. In order to determine the possible involvement of parasite proteases in this process, L. amazonensis promastigotes were collected daily and their proteolytic enzyme patterns analyzed using polyacrylamide gels copolymerized with gelatin. Although promastigotes at different growth stages showed differences in protease patterns, these changes did not correlate with their susceptibility to complement. The major protease of promastigotes, gp63, was expressed at the same level throughout culture, regardless of the complement resistance of the promastigotes. Furthermore, inhibitors specific for the classes of proteases found in L. amazonensis promastigotes did not interfere with the complement-mediated killing of promastigotes. We also investigated the binding of natural antibodies to promastigotes at different stages of growth using ELISA. Although complement-sensitive promastigotes bound significantly more antibodies from fresh normal human serum than complement-resistant promastigotes, equivalent amounts of C3 were detected on their surfaces following complement activation. Moreover, serum depleted of anti-Leishmania antibodies was as efficient in killing promastigotes as the intact serum. These data suggest that the resistance of L. amazonensis to complement killing involves strategies other than that of the regulated expression endogenous proteases capable of inactivating complement components, or the differential ability to bind natural antibodies that might interfere with complement deposition on the parasite surface.