Association between productive parameters and intestinal histomorphological findings in broilers supplemented with probiotics (Saccharomyces cerevisiae, Lactobacillus acidophilus and Bacillus subtilis) / Asociación entre parámetros productivos y hallazgos histomorfológicos en pollos de engorde suplementados con probióticos (Saccharomyces cerevisiae, Lactobacillus acidophilus y Bacillus subtilis)

SUMMARY: This study evaluates the effect of probiotics Saccharomyces cerevisiae, Lactobacillus acidophilus and Bacillus subtilis on production parameters and intestinal histomorphology of broilers of 45 days of age. Eleven 45-day-old Ross 500 broilers were used and classified as control group (CG) (n = 5) or supplemented with probiotics group (n = 8). Histopathological evaluation of duodenum, ileum, and jejunum was performed. The area of the villi height, base and apex were evaluated as well as the size and number of crypts. In addition, mucus production was quantified in different portions of the small intestine. The villi present duodenum of broilers supplemented with probiotics had a greater area (p = 0.0127), a greater basal width (p = 0.0049) and a greater apical width (p = 0.0024), as well as a greater crypt area (p = 0.0189). Significantly higher levels of mucus were noted in the duodenum (p = 0.0480) and jejunum (p = 0.0480) of broilers supplemented with probiotics. We suggest that probiotic supplementation improve the intestinal nutrients absorption.

RESUMEN: Este estudio evalúa el efecto del uso de probióticos como: Saccharomyces cerevisiae, Lactobacillus acidophilus, Bacillus subtilis en parámetros productivos e histomorfologia intestinal de pollos de engorde de 45 días de edad. Fueron usados 11, los cuales fueron clasificados en grupo control (CG) (n = 5) y grupo suplementado con probióticos (PG) (n = 8). Fue realizado análisis histopatológico de secciones de duodeno, íleon y yeyuno. Fue evaluado ancho, altura y área del ápice de la vellosidad, área y número de criptas. Además, fue estimada la producción de moco en los diferentes segmentos del intestino delgado. Fue observada mayor área de la vellosidad en duodeno, PG (p = 0.0127), ancho basal mayor en PG (p = 0.0049) ancho apical mayor en PG (p = 0.0024), así como mayor área de criptas en PG (p = 0.0189). No fueron encontradas diferencias significativas respecto a los segmentos de yeyuno e íleon. PG presentó mayor producción de moco en duodeno (p = 0.0480) y en yeyuno (p = 0.0480). Concluimos que la suplementación con probióticos en pollos de engorde genera cambios en la histomorfologia intestinal, evidenciables en áreas apicales y basales de las vellosidades intestinales. Soporte financiero: Dirección General de Investigaciones - Universidad de los Llanos.

Microencapsulation of Bifidobacterium animalis subsp. lactis and Lactobacillus acidophilus in cocoa butter using spray chilling technology

In the present study, the cells of Bifidobacterium animalis subsp. lactis (BI-01) and Lactobacillus acidophilus (LAC-04) were encapsulated in cocoa butter using spray-chilling technology. Survival assays were conducted to evaluate the resistance of the probiotics to the spray-chilling process, their resistance to the simulated gastric and intestinal fluids (SGF and SIF), and their stability during 90 days of storage. The viability of the cells was not affected by microencapsulation. The free and encapsulated cells of B. animalis subsp. lactis were resistant to both SGF and SIF. The micro-encapsulated cells of L. acidophilus were more resistant to SGF and SIF than the free cells; the viability of the encapsulated cells was enhanced by 67%, while the free cells reached the detection limit of the method (10³ CFU/g). The encapsulated probiotics were unstable when they were stored at 20 °C. The population of encapsulated L. acidophilus decreased drastically when they were stored at 7 °C; only 20% of cells were viable after 90 days of storage. The percentage of viable cells of the encapsulated B. animalis subsp.lactis, however, was 72% after the same period of storage. Promising results were obtained when the microparticles were stored at -18 °C; the freeze granted 90 days of shelf life to the encapsulated cells. These results suggest that the spray-chilling process using cocoa butter as carrier protects L. acidophilus from gastrointestinal fluids. However, the viability of the cells during storage must be improved.

Rapid restoration of colonic goblet cells induced by a hydrolyzed diet containing probiotics in experimental malnutrition

PURPOSE: To investigate the effects of the addition of probiotic bacteria to a hydrolyzed diet on the recovery of goblet cells during renutrition in an animal model of malnutrition. METHODS: Twenty-six male Wistar rats (200-250g) were included in the study. Six were kept under normal conditions (sham group) while twenty received an aproteic diet for 15 days, and were randomized thereafter to receive a hydrolyzed diet containing (n=6; probiotic group) or not (n=6; hydrolyzed group) probiotics (10(6) cfu/g of Streptococcus thermophilus e Lactobacillus helveticus); or immediately killed (n=8; aproteic group). Histological slides containing cecal and sigmoid biopsies were used to counting the number of goblet cells and the goblet cells/colonocytes ratio. RESULTS: Malnutrition diminished the population of goblet cells in all sites. Goblet cells/colonocytes ratio of the probiotic group was significantly greater than hydrolyzed group at the ceccum (0.39 ± 0.03 vs. 0.34 ± 0.02; p=0.02). Only rats fed with probiotics showed complete restoration of the normal goblet cells/colonocytes ratio at the sigmoid (0.37 ± 0,02 vs. 0.22 ± 0,03; p<0,001). CONCLUSION: Streptococcus thermophilus and Lactobacillus helveticus added to a renutrition diet enhance the recovery of mucocal atrophy induced by malnutrition and especially induce a rapid restoration of goblet cells population in the malnourished colonic mucosa.

OBJETIVO: Investigar os efeitos da adição de probióticos em uma dieta hidrolisada na recuperação de células caliciformes durante a renutriçao em um modelo animal de desnutrição. MÉTODOS: Vinte e seis ratos Wistar (200-250g) foram incluídos no estudo. Seis foram mantidos em condições normais (grupo sham) enquanto que 20 receberam uma dieta aproteica por 15 dias, e foram randomizados para receber uma dieta hidrolisada com (n=6; grupo probiótico) ou sem (n=6; grupo hidrolisado) probióticos (10(6) cfu/g of Streptococcus thermophilus e Lactobacillus helveticus); ou foram sacrificados imediatamente (n=8; grupo aproteico). Cortes histológicos contendo biopsias do ceco e sigmoide foram examinados e o número de células caliciformas e a razão caliciformes/colonócitos foram contados. RESULTADOS: A desnutrição diminuiu o número de células caliciformes em todo o cólon. A razão células caliciformes/colonócitos do grupo probiótico foi significantemente maior que o do grupo hidrolizado no ceco (0.39 ± 0.03 vs. 0.34 ± 0.02; p=0.02). Somente os ratos alimentados com probióticos mostrou restauração completa da relação células caliciformes/colonócitos no sigmóide (0.37 ± 0,02 vs. 0.22 ± 0,03; p<0,001). CONCLUSÃO: Streptococcus thermophilus and Lactobacillus helveticus adicionados a uma dieta de renutrição melhora a recuperação da atrofia mucosa induzida pela desnutrição e especialmente induzem a uma rápida restauração da população de células caliciformes na mucosa colônica desnutrida.

In vitro comparative kinetics of adhesive and haemolytic potential of T. vaginalis isolates from symptomatic and asymptomatic females.

The host parasite relationship and pathogenic mechanisms of the commonly reported sexually transmitted urogenital disease, trichomoniasis, are poorly understood. This study was planned to correlate the adhesion properties of Trichomonas vaginalis isolates from symptomatic and asymptomatic women to vaginal epithelial cells in vitro (in presence and absence of L. acidophilus) and to ascertain the haemolytic activity of the isolates, in order to assess these properties as possible markers of pathogenicity. Cytoadherence assay study shows the significant difference in adhesion only up to first 15 minutes of incubation in symptomatic versus asymptomatic isolates. The presence of L. acidophilus was found to be more effective in enhancing the attachment of T. vaginalis in a time dependent manner mostly operative through its pH lowering effect, whereas the excretory secretory products of L. acidophilus reduced the attachment in case of both symptomatic and asymptomatic isolates. Amount of haemoglobin released by isolates from symptomatic patients was significantly higher than by the isolates from asymptomatic women. This investigation forms the basis for future studies to explore the role of other known virulence factors of T. vaginalis in initiation and persistence of vaginal infection by the parasite.

Inhibitory effect of Lactobacillus acidophilus on Helicobacter pylori in peptic ulcer patients: in vitro study.

Lactobacillus acidophilus (L. acidophilus) have been introduced into many fermented dairy products. The presence of L. acidophilus appears to decrease Helicobacter pylori (H. pylori) density in the human stomach and could enhance antibiotic therapy for H. pylori eradication. This study was designed to determine the optimal density of L. acidophilus that has a maximum inhibitory effect on H. pylori in peptic ulcer patients. To determine whether L. acidophilus has an inhibitory effect on H. pylori isolated from peptic ulcer patients and to determine whether the optimal density of L. acidophilus has a maximum inhibitory effect on H. pylori isolated from peptic ulcer patients. H. pylori was isolated from gastric biopsy specimens of peptic ulcer patients. The suspension of pure H. pylori colonies were inoculated into the broth and adjusted to match the density of No.3 MacFarland standard (approximately 9x10(8) cells/ml). Forty microliters of the suspension were equally spread onto each quadrant of the plate and left to dry. L. acidophilus was prepared from LC-1 (Nestle Research Center, Switzerland). They were grown on blood agar and incubated overnight at 37 degrees C. The suspension of L. acidophilus was inoculated into the broth and adjusted to match the density of No.1, No.2, No.3 and No.4 MacFarland standard (approximately 3x10(8), 6x10(8), 9x10(8) and 12x10(8) cells/ml respectively). Ten microliters of each density of L. acidophilus was dropped onto each quadrant of a previously inoculated H. pylori plate and then the plate was kept under microaerophilic conditions for 72 hours. Inhibition clear halo zone of H. pylori around the colonies of L. acidophilus was interpreted as the inhibitory effect. H. pylori were isolated from gastric biopsy specimens of fifteen peptic ulcer patients (eleven patients with gastric ulcer and four patients with duodenal ulcer). A total of sixty tests of inhibitory effect of L. acidophilus on H. pylori were evaluated in the present study. L. acidophilus had inhibitory effect on H. pylori in 13/15 patients (86.67%) and L. acidophilus in the density of No.3 MacFarland standard had a significantly higher inhibitory effect on H. pylori in the density of No.3 MacFarland than that of No.1 (60% vs 20%; p<0.05), No.2 (60% vs 20%; p<0.05) and No.4 (60% vs 20%; p<0.05) MacFarland standard. In conclusion, L. acidophilus has an inhibitory effect on H. pylori isolated from peptic ulcer patients. Approximately an equal density of L. acidophilus on H. pylori has the most favorable effect. This optimal density of L. acidophilus should have maximum effect on H. pylori clearance and could enhance antibiotic therapy for H. pylori eradication in humans.