Potencial bioterapêutico dos probióticos nas parasitoses intestinais

Probióticos são microrganismos vivos que, se administrados em quantidades adequadas, promovem benefícios à saúde do homem e dos animais. O crescente interesse nos probióticos fundamenta-se em estudos clínicos nos quais a administração desses organismos foi avaliada na prevenção e no tratamento de desordens intestinais e sistêmicas. Os potenciais mecanismos de ação desses microrganismos incluem a exclusão competitiva, a produção de metabólitos com atividade antimicrobiana e a modulação da resposta imune. em algumas circunstâncias clínicas específicas, os benefícios produzidos por esses microrganismos foram amplamente documentados, enquanto que em outras os resultados são contraditórios. No presente artigo de revisão, os probióticos foram abordados considerando-se o potencial bioterapêutico desses microrganismos nas parasitoses intestinais.

Lactic Acid Bacteria Antimicrobial Compounds: Characteristics and Applications

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeito do teor de soro, açúcar e de frutooligossacarídeos sobre a população de bactérias lácticas probióticas em bebidas fermentadas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of consumption of probiotics on presence of enterobacteria in the oral cavity

Clinical benefits of probiotics have been clearly reported in different gastrointestinal disorders, many of them caused by enterobacteria. The oral cavity is a port of entry and can be an important reservoir of these microorganisms. This work evaluated whether consumption of probiotics was able to influence the presence of enterobacteria in the oral cavity and the specific secretory response against these microorganisms. Saliva samples of healthy individuals were collected and plated in Mac-Conkey agar. Carriers of Gram-negative, rod-shaped microorganisms in the oral cavity were selected and instructed to use the probiotic Yakult LB for 20 days. Saliva was then collected and enterobacteria species were identified using the API 20 E system and by ELISA using anti-enterobacteria IgA. The results showed reduction in the prevalence of enterobacteria, but no significant changes in enterobacterial counts (log CFU/mL; p = 0.3457). The species most frequently isolated were Enterobacter cloacae and Klebsiella oxytoca, both before and after probiotic consumption. No significant changes were observed in anti-enterobacteria IgA levels. In conclusion, probiotic consumption had some influence on enterobacterial presence in the oral cavity, but did not affect enterobacterial counts or the specific immune secretory response against them.

Probiotics for broiler chickens in Brazil: systematic review and meta-analysis

This study evaluated the efficacy of probiotic utilization as growth promoters in broiler chicken feeding using systematic literature review and meta-analysis. Thirty-five studies were recovered by the systematic review, 27 of which met the following criteria to be included in the meta-analysis: (1) Brazilian studies published between 1995 and 2005; (2) probiotics administered in the diet without growth promoter; (3) results included performance data with the respective coefficient of variation. Meta-analysis have shown that the probiotics promoted better weight gain and feed conversion than the negative control (no antimicrobial) in the initial phase (1 to 20-28 days); nevertheless, results were similar in the total period (1 to 35-48 days). Weight gain and feed conversion were similar between probiotics and the positive control (with antimicrobial) both in the initial and in the total periods. Viability in the total period improved with the use of probiotics in comparison to the negative or positive controls. Sensitivity analysis showed that the results of meta-analysis were coherent. The funnel plots and the Egger regression method evidenced that the studies published in Brazil do not present biased results. It is possible to conclude that the probiotics are a technically viable alternative to antimicrobial growth promoters in broiler feeding. Nevertheless, further studies are necessary to identify eventual differences among the probiotics commercially available in Brazil.

Efeitos de probióticos sobre a digestibilidade, escore fecal e características hematológicas em cães

Avaliaram-se os efeitos da suplementação de dois tipos de probióticos para cães filhotes, que receberam dois tipos de dieta - de alta e de baixa qualidade -, sobre a digestibilidade dos nutrientes, escore fecal e parâmetros sanguíneos. Foram utilizados 18 animais, distribuídos em três tratamentos. No tratamento 1, controle, os cães receberam somente a ração; no tratamento 2, ração com probiótico 1 (Bifidobacterium) e, no tratamento 3, ração com probiótico 2 (Lactobacillus). O experimento foi dividido em duas fases. Verificaram-se que os valores médios do coeficiente de metabolizabilidade da energia bruta (CMEB) na fase 1, caracterizada pela troca da dieta Super Premium para a dieta Standard, apresentaram resultados significativos (P<0,05), sendo os melhores resultados obtidos nos animais que receberam o probiótico 2. Não houve diferenças significativas (P>0,05) para o escore fecal e para as análises hematológicas.

Avaliações ultrassonográfica e radiográfica dos efeitos da suplementação com dois tipos de probióticos sobre o intestino de cães filhotes

Avaliaram-se os efeitos da suplementação de dois tipos de probióticos para cães filhotes da raça Beagle, que receberam dois tipos de dieta de alta e baixa qualidade, sobre os achados em exames de diagnóstico por imagem. Foram utilizados 18 cães filhotes da raça Beagle, distribuídos em três tratamentos inteiramente ao acaso: tratamento 1, controle (ração); tratamento 2, ração com probiótico 1 (Bifidobacterium); e tratamento 3, ração com probiótico 2 (Lactobacillus). O experimento foi dividido em duas fases, cada uma caracterizada pela mudança da dieta de qualidades diferentes. Ao exame ultrassonográfico, foi avaliada a espessura da parede gastrintestinal, e ao radiográfico, a presença de gás em alças intestinais e o espessamento da parede. Observou-se aumento da espessura da parede das alças intestinais (P<0,05), nos animais dos grupos tratados com probiótico 1 e 2 (5,74mm e 6,01mm, respectivamente) em relação aos do grupo-controle (5,03mm). O uso de probióticos não influenciou a produção de gás intestinal. Não houve diferença (P>0,05) entre os tratamentos quanto ao escore de gás intestinal. A avaliação quantitativa permitiu a determinação de um escore de gás intestinal, de forma não invasiva, por meio do software Image J®. Concluiu-se que o diagnóstico por imagem sustenta a recomendação dessa metodologia como ferramenta auxiliar na avaliação nutricional de dietas para cães.

Utilização de probiótico e de lactulose no controle de hiperamonemia causada por desvio vascular portossistêmico congênito em um cão

O desvio vascular portossistêmico pode ser congênito ou adquirido nos cães. A enfermidade pode levar a alterações neurológicas decorrentes de encefalopatia hepática e a hiperamonenia é um dos mecanismos implicados na fisiopatologia deste quadro. O tratamento clínico visa a reduzir os níveis séricos de amônia com o uso de antibióticos e lactulose. em humanos com hepatopatias, os probióticos podem ser utilizados para reduzir a hiperamonemia. A resposta clínica e laboratorial de um cão com desvio vascular portossistêmico foi demonstrada com a utilização de lactulose e de probiótico, isoladamente e associados, sendo que a melhor evolução foi obtida na terapia conjunta.

Atividade antimicrobiana de Lactobacillus acidophilus, contra microrganismos patogênicos veiculados por alimentos

The antimicrobial activity of a commercial probiotic culture, Lactobacillus acidophilus (La5), was tested against two foodborne pathogens, Escherichia coli and Staphylococcus aureus. The antagonistic effect of the probiotic culture in vitro was performed by applying both Multilayer Agar Plate and Agar Well Diffusion methods. The results indicated that the inhibitory substance present on 72 hours culture broth supernatant was extracellular and diffusible. The incubation period of the lactic acid bacteria on MRS Broth, at 3 7 degrees C in aerobic conditions, for the highest lactic acid production (1,08 g/%) was 72 hours, which gave a minimum pH value of the supernatant (3,90) and the best inhibition results by the Well Diffusion Agar Assay, showing inhibition zone diameters of 14,75mm and 15,0mm for E. coli and S. aureus, respectively. The inhibitor compound was not sensitive to proteolytic enzyme and freezing, but was totally inactivated when the supernatant was neutralized with NaOH 1 N solution. The results suggest that the inhibitory activity was due to the lactic acid concentration and the low pH of the probiotic culture broth.

Demonstration of the cellular viability and safety of Enterococcus faecium CRL 183 in long-term experiments

Enterococcus faecium CRL 183, a strain isolated from NSLAB cheese starter, has been the focus of much research on its potential probiotic capacity, although its survival through the gastrointestinal tract has not been demonstrated so far. In order to determine the capacity of E. faecium CRL 183 to survive such conditions, this strain was administered daily to rats for 30 weeks. The experimental animals were divided into Group I: those that did not receive E. faecium, Group II: those that received a pure culture of E. faecium CRL 183 and Group III: animals that received E. faecium CRL 183 in the form of a fermented soy-based product. Faecal samples were collected at the beginning and at the 50%, 75% and 100% stages of the experimental period. Isolation and counts of Enterococcus were carried out on KF selective media. To distinguish the various Enterococcus species in the faeces, biochemical (API Strep 20) and molecular (PCR) tests were performed. Initially, E. faecium was absent from the intestinal flora of the rats; however, after 15 weeks of administration, E. faecium could be recovered from the faeces of Groups II and III, demonstrating that E. faecium CRL 183 was able to survive gastrointestinal transit under the study conditions. This is further evidence of the probiotic qualities of this strain. The safety of the strain was also investigated with regard to body weight and serum biochemical analysis.

Ability of bacteriophages isolated from different sources to reduce Salmonella enterica serovar enteritidis in vitro and in vivo

Salmonella enterica serovar Enteritidis-lysing bacteriophages isolated from poultry or human sewage sources were used to reduce Salmonella Enteritidis in vitro and in experimentally infected chicks. Cocktails of 4 different bacteriophages obtained from commercial broiler houses (CB4O) and 45 bacteriophages from a municipal wastewater treatment plant (WT45O) were evaluated. In experiment 1, an in vitro crop assay was conducted with selected bacteriophage concentrations (105 to 101 pfu/mL) to determine ability to reduce Salmonella Enteritidis in the simulated crop environment. Following 2 h at 37 degrees C, CB40 or WT45O reduced Salmonella Enteritidis recovery by 1.5 or 5 log, respectively, as compared with control. However, CB40 did not affect total SE recovery after 6 h, whereas WT45O resulted in up to a 6-log reduction of Salmonella Enteritidis. In experiment 2, day-of-hatch chicks were challenged orally with 3 x 103 cfu /chick Salmonella Enteritidis and treated cloacally with 1 X 109 WT45O pfu/chick I h postchallenge. One hour later, chicks were treated or not with a commercially available probiotic (Floramax-B11). Both treatments significantly reduced Salmonella Enteritidis recovery from cecal tonsils at 24 h following vent lip application as compared with controls, but no additive effect was observed with the combination of bacteriophages and probiotic. In experiment 3, day-of-hatch chicks were challenged orally with 9 x 103 cfu/chick Salmonella Enteritidis and treated via oral gavage with I X 108 CB40 pfu/chick, 1.2 x 108 WT45O pfu/chick, or a combination of both, I h postchallenge. All treatments significantly reduced Salmonella Enteritidis recovered from cecal tonsils at 24 h as compared with untreated controls, but no significant differences were observed at 48 h following treatment. These data suggest that some bacteriophages can be efficacious in reducing SE colonization in poultry during a short period, but with the bacteriophages and methods presently tested, persistent reductions were not observed.

Ileal and caecal microbial populations in broilers given specific essential oil blends and probiotics in two consecutive grow-outs

Specific essential oil (EO) blends and probiotics used as feed additives have been shown to promote healthy digestive microbials resulting in improved poultry production. Two consecutive experiments were conducted with broilers fed corn-soybean meal diets to determine comparative effects of feed additives on ileal and caecal microbial populations (MP). Ross 708 broilers were placed in 84 pens with previously used litter and treatments maintained in the same pens for both experiments. Eight treatment groups were fed diets containing: Bacitracin methylene disalicylate (BMD) as positive control (PC); no additives as negative control (NC); three probiotics: BC-30; BioPlus 2B (B2B); and Calsporin; and the essential oil blends Crina Poultry Plus (CPP) at 300 or 150 ppm in the first experiment; and CPP at 300 ppm and Crina Poultry AF at 100 ppm in experiment 2. Starter and grower diets contained the ionophore (Coban). Ileal and caecal samples were collected at 43 days of age from male broilers. The DNA of microbial populations was isolated from digesta samples and analysed by denaturing gradient gel electrophoresis to generate percentage similarity coefficients (%SC) from band pattern dendrograms. Differences were observed in ileal and caecal populations depending on treatment, respectively, and especially between experiments. Broilers fed diets with probiotics had very similar MP. The EO CPP at 300 ppm resulted in ilea! MP similar to those observed in chickens fed probiotics. We concluded that antibiotic treatment affected ileal, but no caecal MP. More pronounced changes in ileal and caecal MP were seen in broilers at 43 days of age following probiotic and essential oil treatments.

Using Efinol (R) L during transportation of marbled hatchetfish, Carnegiella strigata (Gunther)

The objective of this experiment was to test the efficacy of a probiotic (Efinol (R) L) during transportation of marbled hatchetfish, Carnegiella strigata. Wild specimens were captured from a small stream and transported for 24 h in plastic fish boxes with a probiotic (10 mg L-1) and probiotic-free water. The boxes were sampled at 3. 12 and 24 h of transport. At the end of the experiment, the survival rate was close to.100%) in both treatments. Dissolved oxygen diminished with time in both treatments, but the probiotic group had significantly higher levels. Conductivity. pH and ammonia increased significantly during the transport. demonstrating higher levels in the probiotic-free group. Fish from both treatments presented very high net Na+ and K+ effluxes after 3 h of transport. At 24 h, net K+ effluxes in fish of the probiotic treatment reached values close to zero and a significantly lower Na+ efflux was observed. Cortisol levels in both treatments at 3 and 12 h were significantly higher than that in control samples. Higher body cortisol levels were observed in the probiotic-frec group than that in the probiotic group at 3 and 12 h. The results demonstrate that addition of a probiotic during fish transport improves water quality and leads to fish presenting a lower stress response intensity.

Detection of T lymphocytes in intestine of broiler chicks treated with Lactobacillus spp. and challenged with Salmonella enterica serovar enteritidis

The expression of immune response in the form of leukocytic infiltrate by CD3+, CD4+, and CD8+ cells in the epithelium and in the intestinal lamina propria of chicks was studied in the present work by means of immunohistochemical reaction. The chicks were treated with Lactobacillus spp. or cecal microflora (CM) and experimentally challenged or not with Salmonella enterica serovar Enteritidis. The 320 birds utilized were divided into 4 groups containing 80 chicks each and submitted to treatments with Lactobacillus reuteri, Lactobacillus salivarius, Lactobacillus acidophilus, and CM. Each group was subdivided into 4 subgroups of 20 birds each and classified into a subgroup that did not receive treatment (negative control), subgroup treated, subgroup treated and challenged with Salmonella Enteritidis, and subgroup only challenged with Salmonella Enteritidis (positive control). The results obtained show that the treatment with L. reuteri, L. salivarius, L. acidophilus, or CM and challenged or not with Salmonella Enteritidis determine immune response in the form of leukocytic infiltrate by CD3+ and CD8+ lymphocytes followed by CD4+ in the epithelium and in the lamina propria of the duodenum, jejunum, and cecum of chicks up to 12 d of age. The quantity of CD3+ lymphocytes was significantly higher (P < 0.05) in the intestine of chicks treated with L. acidophilus or CM and challenged or not with Salmonella Enteritidis; however, the higher quantity of CD8+ lymphocytes was in the intestine of chicks treated with CM and challenged with Salmonella Enteritidis. The duodenum was the segment in which the immune response by T cells (CD3+, CD4+, and CD8+) was stimulated with the greatest intensity, followed by, respectively, the jejunum and cecum. The quantity of CD3+ lymphocytes present in the duodenum, jejunum, and cecum increases with the age of chicks, independent of the stimulus determined by treatments or challenge.