Comparison of two oral probiotic preparations in a randomized crossover trial highlights a potentially beneficial effect of Lactobacillus paracasei NCC2461 in patients with allergic rhinitis.

BACKGROUND: There is promising but conflicting evidence to recommend the addition of probiotics to foods for prevention and treatment of allergy. Based on previous studies with fermented milk containing Lactobacillus paracasei NCC2461, we aimed to compare the effect of a powder form of the latter probiotic with the effect of a blend of Lactobacillus acidophilus ATCC SD5221 and Bifidobacterium lactis ATCC SD5219 in patients with allergic rhinitis. METHODS: A double-blind, randomized, cross-over study, involving 31 adults with allergic rhinitis to grass pollen, was performed outside the grass pollen season (registration number: NCT01233154). Subjects received each product for 4-weeks in two phases separated by a wash-out period of 6 to 8 weeks. A nasal provocation test was performed before and after each 4-week product intake period, and outcome parameters (objective and subjective clinical symptoms; immune parameters) were measured during and/or 24 hours after the test. RESULTS: Out of the 31 subject enrolled, 28 completed the study. While no effect was observed on nasal congestion (primary outcome), treatment with NCC2461 significantly decreased nasal pruritus (determined by VAS), and leukocytes in nasal fluid samples, enhanced IL-5, IL-13 and IL-10 production by peripheral blood mononuclear cells in an allergen specific manner and tended to decrease IL-5 secretion in nasal fluid, in contrast to treatment with the blend of L. acidophilus and B. lactis. CONCLUSIONS: Despite short-term consumption, NCC2461 was able to reduce subjective nasal pruritus while not affecting nasal congestion in adults suffering from grass pollen allergic rhinitis. The associated decrease in nasal fluid leukocytes and IL-5 secretion, and the enhanced IL-10 secretion in an allergen specific manner may partly explain the decrease in nasal pruritus. However, somewhat unexpected systemic immune changes were also noted. These data support the study of NCC2461 consumption in a seasonal clinical trial to further demonstrate its potentially beneficial effect.

IgA production, coliforms analysis and intestinal mucosa morphology of piglets that received probiotics with viable or inactivated cells

Two types of probiotics were used in piglets. One product is a mixed culture of viable Lactobacillus acidophilus, Enterococcus faecium e Bifidobacterium bifidum. The second product is composed of inactivated Lactobacillus acidophilus cells. The piglets received two weekly oral doses for 30 days while a control group did not receive probiotics. All piglets were euthanized at the 30th day of life and the mesenteric lymph nodes, the small intestine, and blood samples were collected. The tissue samples were studied by light microscopy and the blood serum was analyzed by ELISA method. The treatment with the probiotic with viable cells produced higher serum levels of IgA (P<0.05) and more IgA expressing cells were found in the mesenteric lymph nodes than observed in the inactivated cells treatment or control groups (P<0.05). Also, intestinal villi were longer, crypts were deeper (P<0.05) and fecal coliform count was lower than found in the inactivated product (P<0.05). These results suggest that viable probiotics are more efficient than inactivated probiotics to induce immunostimulation and intestinal modifications in piglets, thus improving their health and development.

Uso de probiótico sobre a ativação de células T e controle de Salmonella Minnesota em frangos de corte

Para avaliar o efeito do probiótico sobre a resposta imunológica de frangos de corte desafiados com Salmonella Minnesota (SM), 60 frangos foram divididos em três grupos: CN- (controle negativo) aves que não foram inoculadas com SM, CP- (controle positivo) aves inoculadas com SM e Probiótico- aves suplementadas na ração com probiótico composto de Lactobacillus acidophilus, L. plantarium, L. rhamnosus, L. bulgaricus, Enterococcus faecium, Streptococcus thermophilus e Bifidobacterium bifidum e desafiadas com SM. Aos 14 dias foi realizada a inoculação com SM e aos 7 e 35 dias foram quantificadas células caliciformes, CD4+ e CD8+ na mucosa intestinal do íleo e ceco. Aves suplementadas com probióticos aos 7 dias de idade apresentaram aumento significativo (P≤0,05) de células caliciformes e CD4+ no íleo e de células CD8+ no ceco. Aos 35 dias houve aumento significativo (P≤0,05) das células CD8+ nas aves inoculadas do CN e Probiótico. A utilização de probióticos proporcionou redução significativa (P≤0,05) da contagem de Salmonella sp.

Does the Ingestion of Isoxaflutole Herbicide Affect the Midgut and Salivary Glands of Pentatomidae Predators?

Podisus nigrispinus is a generalist predator naturally occurring in agricultural and forestry systems that effectively contributes to the population balance of phytophagous insects, especially defoliating caterpillars. Histological changes were evaluated in the salivary glands and midgut of P. nigrispinus caused by ingestion of systemic herbicide isoxaflutole. These predator females were fed with leaves of eucalyptus plants, Tenebrio molitor pupae or water, contaminated or not by herbicide. Salivary glands and midguts were dissected, processed and analyzed under a light microscope. Activity level and cell morphology of the salivary glands and midgut showed differences among insects fed on plants, contaminated water or pupae. The epithelia of the salivary gland and midgut of individuals which had no contact with the herbicide showed homogeneous cytoplasm, nucleus with predominance of decondensed chromatin and evident nucleoli, intense cell activity features. As for the insects in contact with contaminated food, they presented undeveloped nucleus and condensed chromatin. The luminal contents of the salivary glands in the contaminated insects had become more acidophilus than in insects without poisoning, as well as having heterogeneous and granular secretion, being more evident in the bioassay in which the insects fed on contaminated water. There was a marked morphological change in the midgut cells in contaminated insects. High degree of apoptosis, disorganization and secretory vacuoles in the epithelial cytoplasm were observed. The apical portion of the midgut cells proved undeveloped, irregular and partially destroyed. It is concluded that isoxaflutole causes morphological changes in the digestive system of the predator P. nigrispinus.

Efeito de probiótico na modulação dos níveis de colesterol sérico e no peso do fígado de ratos alimentados com dieta rica em colesterol e ácido cólico

Alimentos contendo probióticos são conhecidos como funcionais e têm sido recomendados para indivíduos com hipercolesterolemia. Com o objetivo de avaliar seu efeito na modulação dos níveis de colesterol sérico, foi conduzido um estudo de 28 dias, utilizando-se 90 ratos machos Wistar, distribuídos entre os tratamentos: Padrão, com 30 animais e Controle, LDR (Leite Desnatado Reconstituído) e P (Probiótico), com 20 animais cada. O grupo Padrão recebeu a dieta AIN-93G durante todo o período experimental. Os demais grupos receberam a mesma dieta acrescida de 1% de colesterol e 0,1% de ácido cólico. Do 15º ao 28º dia, o grupo LDR recebeu 0,1mL/dia/animal de leite desnatado reconstituído a 10% de sólidos não gordurosos e o grupo P, recebeu 0,1mL/dia/animal de probiótico contendo 10(10) UFC/mL de Lactobacillus acidophilus NCFM. Avaliou-se o colesterol total, LDL-colesterol e HDL-colesterol e o peso do fígado, imediatamente após o término da administração do probiótico, onde os ratos tiveram o fígado retirado e o sangue coletado por punção cardíaca. A adição de colesterol e ácido cólico à dieta não elevou (p>0,05) os níveis de colesterol do grupo Controle, LDR e P em relação ao grupo Padrão, no entanto, promoveu aumento significativo de peso (p<0,05) do fígado e alteração da coloração normal do mesmo (de vermelho vivo para vermelho pálido) devido à deposição de gordura neste órgão. Assim, a dieta rica em colesterol e ácido cólico não promoveu hipercolesterolemia nos animais, mas induziu fígado gorduroso nos mesmos. Em conseqüência, o consumo de L. acidophilus não alterou (p>0,05) os níveis de colesterol sérico dos animais, uma vez que estes níveis se mantiveram normais durante todo o período experimental. Observou-se também que a administração de probiótico não impediu o acúmulo de gordura no fígado dos animais que receberam dieta rica em colesterol e ácido cólico.

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.

Textura instrumental de queijo petit-suisse potencialmente probiótico: influência de diferentes combinações de gomas

O efeito de diferentes combinações de gomas sobre os parâmetros de textura instrumental de queijo tipo petit-suisse probiótico foi avaliado. Queijos petit-suisse foram produzidos utilizando-se massa-base de queijo quark preparada com o starter Streptococcus thermophilus e os probióticos Lactobacillus acidophilus e Bifidobacterium longum. Três formulações de queijo petit-suisse foram preparadas, a partir de massa-base de queijo quark adicionada de 0,75% do produto final da mistura de gomas - xantana (X), carragena (C), guar (G), pectina (P): F1 = 2,5X:2,5C:5G; F2 = 2X:3C:5P; F3 = 5C:5G. Os parâmetros avaliados após 1, 7, 14 e 21 dias de armazenamento do produto a 4±1ºC incluíram a contagem de probióticos, a textura instrumental, o pH e a umidade. As populações de probióticos mantiveram-se acima de 6,40 log UFC/g para L. acidophilus e de 7,30 log UFC/g para B. longum. Houve diferença significativa entre as formulações (p<0,05) para todos os parâmetros de textura, exceto a firmeza de F1. O pH e a umidade apresentaram comportamento semelhante para as três formulações. Dentre as formulações estudadas, F1 foi considerada a melhor, por sua firmeza mais estável durante o período de armazenamento.

Efeito prebiótico do mel sobre o crescimento e viabilidade de Bifidobacterium spp. e Lactobacillus spp. em leite

Para ser considerado prebiótico, um microrganismo deve atender a uma série de requisitos, sendo a manutenção da viabilidade, um dos principais. Culturas probióticas de Lactobacillus spp. e Bifidobacterium spp. foram cultivadas em leite em pó desnatado reconstituído 12% adicionado de 3% (p/v) de mel pasteurizado. Foram preparados controles sem mel. Todos os cultivos mantiveram-se viáveis por 46 dias a 7 °C atendendo o número mínimo exigido pela legislação. O maior número de células viáveis de L. casei-01 e L. casei Shirota (>9,0 log10 UFC.mL-1) foi observado nos cultivos contendo mel. A acidez titulável produzida por estas culturas foi de 1,44%. O número de células viáveis de L. acidophilus Sacco® aos 46 dias em cultivos com mel foi significativamente maior (p < 0,05) que nos controles. Considerando todo o período de estocagem, o mel exerceu efeito positivo significativo (p < 0,05) apenas para as culturas de Bifidobacterium. O menor crescimento e a menor acidificação no 46º dia foram observados nos cultivos de Bf. lactis Bb-12, sendo de 7,63 log10 UFC.mL-1 e 0,61% de acidez em presença de mel e 6,11 log10 UFC.mL-1 e 0,30% de acidez nos controles. Diferentemente, os cultivos de Bf. lactis Sacco® atingiram contagens de 9,11 log10 UFC.mL-1 e produziram 1,11% de acidez.

The influence of different combinations of probiotic bacteria and fermentation temperatures on the microbiological and physicochemical characteristics of fermented lactic beverages containing soybean hydrosoluble extract during refrigerated storage

Lactic beverages containing probiotics were prepared with whole UHT milk, whey of Mozzarella cheese, soybean hydrosoluble extract and sugar. Three formulations were studied, each one containing a different combination of probiotic/starter bacteria, fermented at two different temperatures (37 and 45 °C). The aim of this work was to verify the influence of these variables on the viability of probiotic microorganisms and on the physicochemical stability of lactic beverages during storage under refrigeration (21 days at 7 °C). The results indicated that the fermentation temperature had a significant effect on the viability of probiotic bacteria. Counts for Lactobacillus acidophilus were affected by storage time, resulting appropriate after 21 days only for the beverage fermented at 37 °C. Physicochemical parameters did not exhibit drastic variations - proving the stability of formulations during storage. Cells of Bifidobacterium spp. showed high survival ability, probably due to the presence of growth promoters from soybean and cheese whey. The fermentation temperature of 37 °C allowed counts above the minimum limit for all the studied microorganisms, being preferred to the temperature of 45 °C. The inclusion of soybean hydrosoluble extract, a prebiotics source, resulted in a symbiotic product with more benefits to the health of consumers.

Phisicochemical, sensory, and microbiological evaluation and development of symbiotic fermented drink

The goal of this study was to develop a symbiotic lacteous drink, evaluate its physicochemical and sensory characteristics, and verify the viability of Lactobacillus acidophilus in the drink. The milk serum-based drink consisted of 50% milk serum containing 10% saccharose, 25% powdered milk, 15% yacon pulp, and cultures of Lactobacillus acidophilus-La 5E and Bifidobacterium bifidum BB12. It was stored for up to 21 days under refrigeration. The milk serum-based drink was analyzed for protein, ether extract, total dietary fiber, total frutans, carbohydrate content, color, pH, acidity, and contamination by coliforms, and Salmonella sp. Coliforms and Salmonella sp were not detected, and L. acidophilus and B. bifidum provided satisfactory probiotic counts up to 21 days of storage under refrigeration. Lactobacillus acidophilus resistance to gastric acids and bile salts was detected only up to seven days of storage when evaluated in vitro. Sensory analysis and purchase viability were evaluated by consumers at 0, 7, 14, and 21 days of storage. Based on the analytical results and consumer evaluation, the drink was acceptable up to seven days of storage.

Probiotic Effects of Lactic Acid Bacteria Against Vibrio Alginolyticus in Penaeus (Fenneropenaeus) Indicus (H. Milne Edwards)

Cell free extracts of four strains of Lactic acid bacteria (LAB) viz. Lactobacillus. acidophilus, Streptococcus.cremoris, Lactobacillus bulgaricus –56 and Lactobacillus bulgaricus –57 inhibited growth of Vibrio alginolyticus in nutrient broth. The antagonism of LAB to Vibrio alginolyticus was further confirmed by streak plating wherein suppression of growth of Vibrio was obtained. Juveniles of Penaeus indicus (average weight 0.985 ± 0.1 g) on administering orally a moist feed base containing 5 × 106 cells·g of the four LAB probionts for a period of four weeks showed better survival (56 to 72%) when challenged with V. alginolyticus by intra-muscular injection of 0.1 ml containing 3 × 109 cells·ml. Animals maintained on a diet devoid of bacterial biomass exhibited 80% mortality. No external or internal pathological changes were observed in shrimp fed with the LAB incorporated diets. Results showed inhibition of V. alginolyticus by LAB and stimulation of the non-specific immune response resulting in resistance to disease in the shrimp fed on LAB incorporated diets.

In vitro evaluation of the fermentation properties of galactooligosaccharides synthesised by alpha-galactosidase from Lactobacillus reuteri

Stirred, pH-controlled anaerobic batch cultures were used to evaluate the in vitro utilisation by canine gut microflora of novel alpha-galactooligosaccharides synthesised with an enzyme extract from a canine Lactobacillus reuteri strain. Fructooligosaccharides (FOS), melibiose and raffinose were used as reference carbohydrates for the prebiotic properties of the synthesised oligosaccharide (galactosyl melibiose mixture-GMM). Addition of Lactobacillus acidophilus was used as control for the evaluation of the synbiotic properties of the oligosaccharide with L. reuteri. Populations of predominant gut bacterial groups were monitored over 48 h of batch culture by fluorescent in situ hybridisation, and short-chain fatty acid (SCFA) production was measured. GMM showed a higher increase in bifidobacteria and lactobacilli population number and size as well as a higher decrease in clostridia population number and size compared to the commercial prebiotics (FOS, melibiose, raffinose). This prebiotic effect was further increased by the addition of L. reuteri followed by a change in the SCFA production pattern compared to GMM alone or GMM with L. acidophilus. The observed change in SCFA production was in accordance with the fermentation properties of L. reuteri, suggesting that the novel synbiotic had a significant effect on the canine gut microflora fermentation.

Modulation of anti-pathogenic activity in canine-derived Lactobacillus species by carbohydrate growth substrate

Aims: To investigate the effect of various carbon sources on the production of extracellular antagonistic compounds against two Escherichia coli strains and Salmonella enterica serotype Typhimurium by three canine-derived lactobacilli strains. Methods and Materials: Cell-free preparations, pH neutralized, were used in antibiotic disc experiments as an initial screening. The bacteria/carbohydrate combinations that showed inhibition of the growth of those pathogens, were further investigated in batch co-culture experiments. The cell-free supernatants of the cultures, that decreased the population number of the pathogens in the co-culture experiments to log CFU ml(-1) less than or equal to 4, were tested for inhibition of the pathogens in pure cultures at neutral and acidic pH. Conclusions: The results showed that the substrate seems to affect the production of antimicrobial compounds and this effect could not just be ascribed to the ability of the bacteria to grow in the various carbon sources. L. mucosae, L. acidophilus and L. reuteri, when grown in sugar mixtures consisting of alpha-glucosides (Degree of Polymerization (DP) 1-4) could produce antimicrobial compounds active against all three pathogens in vitro. This effect could not be attributed to a single ingredient of those sugar mixtures and was synergistic. This inhibition had a dose-response characteristic and was more active at acidic pH. Significance and Impact of the Study: Knowledge of the effect that the carbon source has on the production of antimicrobial compounds by gut-associated lactobacilli allows the rational design of prebiotic/probiotic combinations to combat gastrointestinal pathogens.

Probiotics and cancer: from in vitro to human studies

Studies in cell cultures and animal models provide evidence that probiotics can beneficially influence various stages in development of colon cancer including tumor initiation, promotion and metastasis. For example, oral administration of Lactobacillus and Bifidobacterium strains can prevent genotoxic damage to the colonic epithelium (considered to be an early stage of the carcinogenic process). Administration to rats of probiotics reduced the incidence of carcinogen-induced pre-cancerous lesions (aberrant crypt foci) in the colon. Furthermore a combination of Bifidobacterium longum and inulin (a prebiotic) was more effective than either treatment alone. In this latter study, the dietary treatments were given after exposure to the carcinogen, which suggests that the protective effects were being exerted at the promotional phase of carcinogenesis. L. acidophilus feeding has been shown to decrease the incidence of colon tumors in rats challenged with a carcinogen and B. longum reduced the incidence of carcinogeninduced colon, liver and mammary tumors. There is limited evidence from epidemiological studies for protective effects of products containing probiotics in humans, but a number of recent dietary intervention studies in healthy subjects and in polyp and cancer patients have yielded promising results on the basis of biomarkers of cancer risk and grade of colorectal tumors.

Caseinoglycomacropeptide inhibits adhesion of pathogenic Escherichia coli strains to human cells in culture

Caseinoglycomacropeptide (CGMP) derived from kappa-casein was investigated for its ability to inhibit the adhesion of 3 strains of verotoxigenic Escherichia coli (VTEC) and 3 strains of enteropathogenic Escherichia coli (EPEC) to human HT29 tissue cell cultures. Effects on adhesion of Desulfovibrio desulfuricans, Lactobacillus pentosus, Lactobacillus casei, Lactobacillus acidophilus, and Lactobacillus gasseri were also investigated. Generally, CGMP exerted effective anti-adhesive properties at a dose of 2.5 mg/mL, albeit with a high degree of strain specificity. The CGMP reduced adhesion of VTEC strains to < 50% of the control and reduced adhesion of EPEC strains to between 80 and 10% of the control. The CGMP also reduced the adhesion of L. pentosus and L. casei to 44 and 42%, respectively. A slight but significant reduction of L. acidophilus, to 81%, was observed, but no significant effects were detected with either Dsv. desulfuricans or L. gasseri. Further investigation of the dose response relationships with the E. coli strains gave IC50 values ranging between 0.12 and 1.06 mg/mL.