Optimization of Growth Parameters for Enhancing the Production of Biosurfactants from Pediococcus pentosaceus S-2

Numerous Lactic acid bacteria (LAB) have been found to be capable of synthesizing surface-active compounds i.e biosurfactants. These are amphiphilic compounds produced by microorganisms on their cell surface or secreted extracellularly that have a tendency to reduce surface and interfacial tension. In the present study, different process parameters including nitrogen and carbon source, pH, temperature, aeration and agitation were optimized to maximize the production of biosurfactants from Pediococcus pentosaceus S-2. Xylose (1.5%) and yeast extract (1.5%) act as better carbon and nitrogen sources respectively for the production of biosurfactants. Maximum biosurfactant yield was observed at pH 6, a temperature of 35o C, an agitation rate of 200 rpm and with inoculum size of 3%. The high yield of biosurfactants produced from Pediococcus pentosaceus S-2 by utilizing media supplemented with whey under optimized conditions.

Performance of lactating cows fed sugarcane silage treated with microbial additives

Background: No studies have been conducted evaluating sugarcane silage associated with both Lactobacillus plantarum and Pediococcus pentosaceus for lactating dairy cows. Objective: To evaluate diets containing different roughages with and without microbial inoculants on intake, digestibility and milk yield of medium-producing cows. Methods: A total of 15 Holstein cows distributed into a randomized block design were used. Dietary treatments were: 1) a corn silage-based diet (CS), 2) a fresh sugarcane-based diet (SC), 3) a sugarcane silage ensiled without inoculant (SS), 4) sugarcane silage ensiled with Lactobacillus buchneri (SSLB), and 5) sugarcane silage ensiled with Lactobacillus plantarum and Pediococcus pentosaceus (SSLP). Results: Digestible organic matter intake (DOMI) was lower (p<0.05) in cows fed SSLB (9.77 kg day-1) when compared with cows fed CS (13.29 kg day-1) and SSLP (12.42 kg day-1). Ensiling of sugarcane increased intake of neutral detergent fiber (NDF) compared to SC (mean of 6.00 kg day-1 versus 4.97 kg day-1; p<0.05). Dry matter digestibility was greater (p<0.05) in CS (77.80%) compared with diets based on sugarcane silage, whereas NDF digestibility was lower for treatments with sugarcane (p<0.05). Milk yield was similar among CS (27.99 kg), SC (25.59 kg), and silages with additives (25.47 and 27.07 for SSLB and SSLP, respectively). Cows fed CS produced more fat-corrected milk (25.89 kg) than those fed sugarcane-based diets (p<0.05). Conclusions: Fresh sugarcane or sugarcane silage with additives can be used as a roughage source for dairy cows producing up to 23.43 kg d-1 fat-corrected milk, considering the total diet is properly balanced.

Antecedentes: No existen estudios que evalúen la adición de Lactobacillus plantarum y Pediococcus pentosaceus en el ensilaje de caña de azúcar para vacas lactantes. Objetivo: Evaluar dietas con diferentes forrajes y ensilajes con y sin inoculantes microbianos sobre el consumo, digestibilidad y producción lactea de vacas de mediana producción. Metodología: Quince vacas Holstein se distribuyeron en un diseño de bloques al azar. Los tratamientos fueron: 1) dieta con ensilaje de maíz (CS), 2) dieta con caña de azúcar fresca (SC), 3) dieta con ensilaje de caña sin inoculantes (SS), 4) dieta con ensilaje de caña de azúcar ensilada con Lactobacillus buchneri (SSLB), y 5) dieta con ensilaje de caña de azúcar ensilada con Lactobacillus plantarum y Pediococcus pentosaceus (SSLP). Resultados: El consumo de materia orgánica digestible fue menor (p<0,05) en la dieta SSLB (9,77 kg day-1) en comparación con CS (13,29 kg dia-1) y SSLP (12,42 kg dia-1). El ensilaje de caña de azúcar promovió un mayor consumo de fibra detergente neutra (NDF) en comparación con SC (promedio de 6,00 kg dia-1 versus 4,97 kg dia-1; p<0,05). La digestibilidad de la materia seca fue mayor (p<0,05) para la dieta CS (77,80%) en comparación con las dietas con ensilaje de caña de azúcar, mientras que la digestibilidad de la NDF fue menor para las dietas a base de caña de azúcar (p<0,05). La producción de leche fue similar entre CS (27,99 kg), SC (25,59 kg) y ensilajes con aditivos (25,47 y 27,07 para SSLB y SSLP, respectivamente). Las vacas alimentadas con ensilaje de maíz produjeron más leche corregida por grasa (25,89 kg) que las alimentadas con dietas a base de caña de azúcar (p<0,05). Conclusión: La caña de azúcar fresca o ensilada con aditivos se puede utilizar como fuente de forraje para vacas que producen hasta 23,43 kg d-1 leche corregida por grasa, siempre que la dieta total esté equilibrada adecuadamente.

Antecedentes: Há carência de estudos avaliando a adição de Lactobacillus plantarum e Pediococcus pentosaceus na silagem de cana-de-açúcar para vacas em lactação. Objetivo: Avaliar dietas contendo diferentes forragens e silagens com e sem inoculantes microbianos sobre o consumo, digestibilidade e produção de leite de vacas de média produção de leite. Métodos: Quinze vacas Holandesas foram distribuídas em um delineamento em blocos casualizados. Os tratamentos foram: 1) dieta com silagem de milho (CS), 2) dieta com cana-de-açúcar fresca (SC), 3) dieta com silagem de cana ensilada sem inoculantes (SS), 4) dieta com silagem de cana-de-açúcar ensilada com Lactobacillus buchneri (SSLB), ou 5) dieta com silagem de cana- de-açúcar ensilada com Lactobacillus plantarum, and Pediococcus pentosaceus (SSLP). Resultados: O consumo de matéria orgânica digestível foi menor (p<0,05) na dieta SSLB (9,77 kg day-1) comparada com CS (13,29 kg dia-1) e SSLP (12,42 kg dia-1). A ensilagem da cana-de-açúcar promoveu maior consumo de fibra em detergente neutro (FDN) em comparação com SC (média de 6,00 kg dia-1 versus 4.97 kg dia-1; p<0.05). A digestibilidade da matéria seca foi maior (p<0,05) para a dieta CS (77,80%) comparada com as dietas com silagem de cana-de-açúcar, enquanto que a digestibilidade da FDN foi menor para as dietas baseadas em cana-de-açúcar (p<0,05). A produção de leite foi similar entre CS (27,99 kg), SC (25,59 kg) e silagens com aditivos (25,47 e 27,07 para SSLB e SSLP, respectivamente). Vacas alimentadas com silagem de milho produziram mais leite corrigido para gordura (25,89 kg) que aquelas alimentadas com dietas baseadas em cana-de-açúcar (p<0,05). Conclusão: A cana-de-açúcar fresca ou ensilada com aditivos pode ser utilizada como fonte volumosa para vacas produzindo até 23,43 kg d-1 leite corrigido para gordura, desde que a dieta total esteja apropriadamente balanceada.

Probiotic potential of lactic acid bacteria isolated from Vietnamese sour-fermented fish product

Aims@#To isolate and characterize the lactic acid bacteria (LAB) strains from the “mam chua ca ro” (sour fermented fish) in the South of Vietnam and investigate their potential anti-bacterial properties.@*Methodology and results@#Four LAB strains (MCR1, MCR2, MCR3 and MCR4) were isolated from the "mam chua ca ro" product and their anti-bacterial activity was determined using the spot assay and the paper disc diffusion method. The isolated LABs can inhibit Escherichia coli ATCC 25922, Staphyloccocus aureus ATCC 25923 and Vibrio parahaemolyticus BV016 and produce bacteriocin to control the growth of E. coli ATCC 25922 and S. aureus ATCC 25923, except V. parahaemolyticus. MCR2 was chosen to sequence 16S rRNA of Pediococcus acidilactic.@*Conclusion, significance and impact of study@#On the basis of their prominent anti-pathogenic bacteria activity, LAB strains isolated from Vietnamese sour-fermented fish products were verified as prospective probiotics.

Enhanced killing of multidrug-resistant Pseudomonas aeruginosa ATCC10145 through a combined action of antibiotics and bacteriocin from Pediococcus pentosaceus TU2

Aims@#Due to its rapid development of resistance against most conventional antibiotics, there is an urgent need to develop new antimicrobial agents and strategies to overcome the challenges in combating multidrug-resistant Pseudomonas aeruginosa infections. This study aimed to determine the antipseudomonal potency of bacteriocin produced by Pediococcus pentosaceus TU2 when combined with conventional antibiotics.@*Methodology and results@#The checkerboard method and time-kill assay were conducted to investigate the antagonism interaction and kinetics of the bacteriocin TU2 and selected antibiotics against Pseudomonas aeruginosa ATCC10145. The scanning electron microscope (SEM) was used to observe the cell surface morphological changes of the treated P. aeruginosa ATCC10145. The combination of bacteriocin TU2 with ciprofloxacin and tetracycline resulted in a 4-fold reduction in minimum inhibitory concentration (MIC) and a fractional inhibitory concentration index (ΣFICI) of 0.5, indicating a synergistic interaction against P. aeruginosa ATCC10145. Similarly, the time-kill assay showed that the combination of bacteriocins TU2 respectively with chloramphenicol and tetracycline exerted enhanced bactericidal effect at 8 h and 10 h of treatments compared to treatment with antimicrobial agents alone. Results from SEM suggested that bacteriocin TU2 might cause pore formation on cells and thus enhanced the membrane permeability of antibiotics and intensified the membrane leakage that led to cell death of P. aeruginosa ATCC10145.@*Conclusion, significance and impact of study@#The combined antagonistic effect of bacteriocin TU2 and antibiotics could be a promising strategy in combating P. aeruginosa infections and may be applied in therapeutic industries.

Dynamic changes of total acid and bacterial communities during the traditional fermentation of Hong Qu glutinous rice wine

Background: Hong Qu glutinous rice wine (HQGRW) is brewed under non-aseptic fermentation conditions, so it usually has a relatively high total acid content. The aim of this study was to investigate the dynamics of the bacterial communities and total acid during the fermentation of HQGRW and elucidate the correlation between total acid and bacterial communities. Results: The results showed that the period of rapid acid increase during fermentation occurred at the early stage of fermentation. There was a negative response between total acid increase and the rate of increase in alcohol during the early fermentation stage. Bacterial community analysis using high-throughput sequencing technology was found that the dominant bacterial communities changed during the traditional fermentation of HQGRW. Both principal component analysis (PCA) and hierarchical clustering analysis revealed that there was a great difference between the bacterial communities of Hong Qu starter and those identified during the fermentation process. Furthermore, the key bacteria likely to be associated with total acid were identified by Spearman's correlation analysis. Lactobacillus, unclassified Lactobacillaceae, and Pediococcus were found, which can make significant contributions to the total acid development (| r| N 0.6 with FDR adjusted P b 0.05), establishing that these bacteria can associate closely with the total acid of rice wine. Conclusions: This was the first study to investigate the correlation between bacterial communities and total acid during the fermentation of HQGRW. These findings may be helpful in the development of a set of fermentation techniques for controlling total acid.

Pediococcus pentosaceus as probiotic with cholesterol-lowering ability

Aims@#Hypercholesterolemia which is an elevated blood cholesterol level that considered as a major risk factor for cardiovascular disease, which is the leading cause of death in many countries. Therefore, lowering the cholesterol level is important to prevent the disease. Lactic acid bacteria (LAB) group are often used as probiotics for their healthpromotion which include cholesterol-lowering effect. The purpose of this study was to evaluate the potency of Pediococcus pentosaceus as probiotic that could reduce cholesterol. @*Methodology and results@#All P. pentosaceus strains were able to survive in acid conditions and in the presence of 0.3% bile salts. These strains had antimicrobial activity against Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, and Salmonella typhimurium ATCC 14028. The LAB were also sensitive to chloramphenicol and showed autoaggregation and coaggregation ability. Pediococcus pentosaceus E5, E7, and E8 were able to remove cholesterol with the highest activity showed by P. pentosaceus E7 (49.00 ± 2.83%). Dead cells and resting cells of P. pentosaceus E5, E7, and E8 (6-22%) also able to reduce the cholesterol but not as effectively as growing cells. Cholesterol lowering is often associated with bile salt hydrolase (BSH) enzyme activity, however none of the isolates were found BSH positive in this study. @*Conclusion, significance and impact of study@#The present study suggests that P. pentosaceus E7 has beneficial probiotic properties which can be exploited for probiotic product with cholesterol-lowering effect.

Effect of microbial immunostimulants on WSSV infection percentage and the expression of immune-related genes in white shrimp (Litopenaeus vannamei) / Efecto de los inmunoestimulantes microbianos en el porcentaje de infección con WSSV y la expresión de genes relacionados con el sistema inmune del camarón blanco (Litopenaeus vannamei) / Efeito de imunoestimulantes microbianos sobre a porcentagem de infecção por WSSV e a expressão de genes relacionados ao sistema imune em camarões brancos (Litopenaeus vannamei)

Abstract Background: The white spot syndrome virus (WSSV) causes high mortalities in aquaculture. The use of immunostimulants increases animal resistance. Objective: To evaluate the WSSV infection percentage and the immunostimulant effect of lactic acid bacteria and yeast (MI= microbial immunostimulants) on WSSV infected Litopenaeus vannamei. Methods: A bioassay was performed for 33 d, with treatments in triplicate. The MI was added to the feed at 8.5 mg/kg feed and offered to shrimp (9.9 ± 3.1 g) daily, every 2 days, or every 3 days. Shrimp were infected with WSSV at 9 and 19 days. The expression of four immune system-related genes was studied using qRT-PCR. Results: No significant differences were observed in growth and survival among treatments. At the end of the bioassay, WSSV infection percentage (low viral load) decreased 8.3 and 25% in treatments III and IV as compared to the control group. Treatments with MI showed significant differences in the relative expression of LvToll, transglutaminase, and prophenoloxidase genes when MI was offered daily as compared to the control group. The MI did not regulate the expression of the superoxide dismutase gene. The WSSV infection percentage decreased when feed with MI was offered every 3 d. Conclusion: The MI decrease WSSV infection percentage in L. vannamei infected with low viral load when it is offered every three days. The MI up-regulates LvToll, Tgase, and proPO genes when it is offered daily. Further research is needed regarding prophylactic treatment with microbial immunostimulants against WSSV in commercial shrimp farms.

Resumen Antecedentes: El virus del síndrome de la mancha blanca (WSSV) ocasiona alta mortalidad en acuacultura. El empleo de inmunoestimulantes incrementa la resistencia de los animales. Objetivo: Evaluar el porcentaje de infección de WSSV y el efecto inmunoestimulante de bacterias ácido lácticas y levadura (MI= inmunoestimulantes microbianos) en Litopenaeus vannamei infectado con WSSV. Métodos: Se realizó un bioensayo durante 33 días con tratamientos por triplicado. Se agregó MI al alimento (8,5 mg/kg de alimento), suministrandolo al camarón (9,9 ± 3,1 g) diariamente, cada 2, o cada 3 días. Los camarones se infectaron con WSSV a los 9 y 19 días. Se estudió la expresión de cuatro genes relacionados con el sistema inmune utilizando qRT-PCR. Resultados: No hubo diferencias significativas en el crecimiento y la sobrevivencia entre los tratamientos. Al final del bioensayo, el porcentaje de infección de WSSV (baja carga viral) disminuyó un 8,3 y un 25% en los tratamientos III y IV en comparación con el grupo control. Los tratamientos con MI mostraron diferencias significativas en la expresión relativa de los genes LvToll, transglutaminasa, y profenoloxidasa cuando se suministró MI diariamente en comparación con el grupo control. Los MI no regularon la expresión del gen de la superóxido dismutasa. El porcentaje de infección de WSSV disminuyó cuando el alimento con MI se aplicó cada 3 d. Conclusiones: Los MI disminuyen el porcentaje de infección de WSSV en L. vannamei infectado con baja carga viral cuando se aplican cada tres días. Además, los MI causan una sobre-expresión de los genes LvToll, Tgase y proPO cuando se administran diariamente. Se necesita más investigación sobre los inmunoestimulantes microbianos como tratamiento profiláctico contra WSSV en granjas comerciales de camarón.

Resumo Antecedentes: O vírus da mancha branca (WSSV) causa alta mortalidade na aqüicultura e o uso de imunoestimulantes aumenta a resistência em animais. Objetivo: Este trabalho avaliou a prevalência do WSSV e o efeito imunoestimulante de bactérias ácido lácticas e levadura (MI= inmunoestimulantes microbianos) em Litopenaeus vannamei infectado con WSSV. Métodos: Se realizou um bioensayo durante 33 dias com tratamientos por triplicado. Se agregou MI al alimento (8,5 mg/kg feed) e se o fornecimento de camarão (9,9 ± 3,1 g) diariamente, cada 2 e cada 3 d. Los camarones foram infectados com WSSV a los 9 y 19 d. Se estudió a expressão de quatro genes relacionados com o sistema imune, use qRT-PCR. Resultados: No hubo diferencias significativas no crescimento e na supervisão entre os tratamientos. Al final del bioensayo, a prevalência do WSSV diminuiu um 8.3 e não 25% nos tratamentos III e IV em comparação com o grupo de controle. Os tratamentos com MI mostraram diferenças significativas na expressão relativa dos genes. LvToll, transglutaminasa e prophenoloxidase quando se administraron em diários de MI em comparação com o controle de grupo. Los MI não regularam a expressão do genótipo da superóxido dismutasa. A prevalência do WSSV diminuiu quando o alimento com MI se aplicó cada 3 d. Conclusões: Los IM diminuir a prevalência de WSSV em L. vannamei infectado com baixa carga viral quando aplicável a cada três dias. Além disso, os IM causaram uma sobre-expressão dos genes LvToll, Tgase e proPO quando se administran diariamente. Há mais informações sobre imunoestimulantes microbianos como tratamento profiláctico contra WSSV en granjas comerciales de camarón.

Heterogeneity of macrolide-lincosamide-streptogramin phenotype & conjugal transfer of erm(B) in Pediococcus pentosaceus

Background & objectives: Pediococcus pentosaceus has been reported to cause clinical infections while it is being promoted as probiotic in food formulations. Antibiotic resistance (AR) genes in this species are a matter of concern for treating clinical infections. The present study was aimed at understanding the phenotypic resistance of P. pentosaceus to macrolide-lincosamide-streptogramin B (MLSB) antibiotics and the transfer of AR to pathogens. Methods: P. pentosacues isolates (n=15) recovered from fermented foods were screened for phenotypic resistance to MLSBantibiotics using disc diffusion and microbroth dilution methods. Localization and transferability of the identified resistance genes, erm(B) and msr(C) were evaluated through Southern hybridization and in vitro conjugation methods. Results: Four different phenotypes; sensitive (S) (n=5), macrolide (M) (n=7), lincosamide (L) (n=2) and constitutive (cMLSB) (n=1) were observed among the 15 P. pentosaceus isolates. High-level resistance (>256 ?g/ml) to MLSBwas observed with one cMLSBphenotypic isolate IB6-2A. Intermediate resistance (8-16 ?g/ml) to macrolides and lincosamides was observed among M and L phenotype isolates, respectively. Cultures with S phenotype were susceptible to all other antibiotics but showed unusual minimum inhibitory concentration (MIC) values of 8-16 ?g/ml for azithromycin. Southern hybridization studies revealed that resistance genes localized on the plasmids could be conjugally transferred to Enterococcus faecalis JH2-2. Interpretation & conclusions: The study provides insights into the emerging novel resistance patterns in P. pentosaceus and their ability to disseminate AR. Monitoring their resistance phenotypes before use of MLS antibiotics can help in successful treatment of Pediococcal infections in humans.

Bioprodução de pediocina por Pediococcus pentosaceus ET34 em resíduo agroindustrial hidrolisado bagaço de cana-de-açúcar / Bioproduction of Pediocin by Pediococcus pentosaceus ET34 in hydrolyzed sugar cane bagasse agroindustrial residue

Um dos grandes desafios nas indústrias alimentícia, farmacêutica e agropecuária é a busca por novos compostos para substituir os antibióticos. Como possíveis candidatos estão as bacteriocinas para serem utilizados paralelamente aos antibióticos ou até substituí-los. As bactérias ácido-láticas podem produzir substâncias inibitórias semelhantes às bacteriocinas (BLIS - bacteriocin-like inhibitory substances) que possuem efeito bacteriostático ou bactericida contra diferentes grupos de bactéria sendo largamente utilizadas como bioconservantes alimentares. Neste contexto, o objetivo deste trabalho foi o emprego de um resíduo agroindustrial, o hidrolisado de bagaço de cana-de-açúcar, como fonte de carbono em cultivos fermentativos para produção do BLIS pela cepa Pediococcus pentosaceus ET 34. Resultados revelaram que as células de ET34 foram capazes de crescer utilizando este resíduo agroindustrial como fonte de carbono e ensaios utilizando planejamento fatorial demonstraram que a agitação não influencia na produção de BLIS. Ao escalonar o cultivo em biorreatores, foi verificado que tanto o crescimento como a atividade antimicrobiana foram semelhantes aos obtidos em bancada com exceção da condição de 25% (v/v) de HBC (hidrolisado de bagaço de cana) que devido a maior viscosidade do meio, resultou em uma diminuição no crescimento e de produção de BLIS. O BLIS produzido por células de ET34 utilizando o HBC como fonte de carbono foi parcialmente purificado por sulfato de amônio e demonstrou atividade contra Listeria monocytogenes ATCC 934 e Salmonella enterica CECT 724. Desta maneira, pode-se concluir que Pediococcus pentosaceus ET34 foi capaz de crescer em meio contendo HBC como fonte de carbono produzindo BLIS em seu sobrenadante com ação frente a diferentes bactérias patogênicas. A possibilidade de utilizar uma fonte alternativa de carbono pode diminuir o custo de processo consideravelmente. Além disso, ensaios de planejamento fatorial, superfície de resposta e escalonamento em biorreator de bancada indicaram que concentrações baixas de HBC (5-15%, v/v) a 35 °C resultaram na maior produção de BLIS

The great challenge in the food, pharmaceutical and agricultural industries is the search for new compounds to replace antibiotics. Bacteriocins are possible candidates that can be used in parallel with the antibiotics or even to replace them. Lactic-acid bacteria can produce bacteriocin inhibitory substances (BLIS - bacteriocin-like inhibitory substances) that have a bacteriostatic or bactericidal effect against different bacterial species and are widely used as food bioconservatives. In this context, the aim of this work was to use of an agroindustrial waste, hydrolyzed sugarcane bagasse, as a carbon source in fermentative cultures for the production of BLIS by Pediococcus pentosaceus ET 34 strain. Results revealed that ET34 cells were able to grow using this agroindustrial residue as a carbon source, and trials using factorial design showed that agitation did not influence on the production of BLIS. When it was perform cultivation scale up in bioreactors, it was verified that both the growth and the antimicrobial activity were similar to those obtained in the workbench with the exception of the condition of 25% (v/v) of HBC (sugarcane bagasse hydrolyzate) that due to its higher viscosity, resulted in a decrease in growth and BLIS production. BLIS produced by ET34 cells using HBC as a carbon source that was partially purified by ammonium sulfate showed activity against Listeria monocytogenes ATCC 934 and Salmonella enterica CECT 724. Thus, it can be concluded that Pediococcus pentosaceus ET34 was able to grow in medium containing HBC as carbon source producing BLIS in its supernatant with action against different pathogenic bacteria. The possibility of using an alternative carbon source can greatly reduce the process cost. In addition, factorial design, response surface and scale up trials in bench bioreactors indicated that low concentrations of HBC (5-15% v/v) at 35 ºC resulted in higher BLIS production

Optimization of the yield of bacteriocin-like substance (BLIS) produced by Pediococcus pentosaceus and its application as food bioconservative / Otimização do rendimento de substância semelhante a bacteriocina (BLIS) produzido por Pediococcus pentosaceus e sua aplicação como bioconservante de alimentos

Bacteriocins are peptides produced by various species of bacteria, especially lactic acid bacteria (LABs), which exhibit a large spectrum of action against spoilage bacteria and foodborne pathogens. However, when this bacteriocin has not been completely characterized regarding its amino acid and the nucleotide sequences of the corresponding gene, the qualified term bacteriocin-like inhibitory substance (BLIS) is recommended. In order to increase the antimicrobial activity of bacteriocins, the ability of probiotics LABs, such as Pediococcus pentosaceus, to ferment different carbon and nitrogen sources has been studied. For the development of an improved culture medium, carbon and nitrogen sources must be considered as nutrients responsible for cell growth and bacteriocin production. The best condition, after 48 h of cultivation, for growth (3.420 g/L) and for BLIS production by Pediococcus pentosaceus ATCC 43200 was in Man, Rogosa and Sharp (MRS) culture medium supplemented with 1.5% peptone, initial pH 6.0 and under the following culture conditions: anaerobiosis, 30oC and agitation of 200 rpm. Compared with control (MRS without supplement), the growth of Pediococcus was significantly lower (1.995 g/L) as well as it reduced significantly its generation time from 2.05 h (control) to 1.28 h (MRS supplemented), a reduction of approximately 62.5%. Moreover, addiction of peptone to MRS medium promoted reduction of 4 h to the Pediococcus exponential phase onset. Regarding BLIS antimicrobial activity, addition of nitrogen source to MRS medium was also quite significant. Through the agar diffusion method, BLIS showed inhibition halos between 12.50 and 19.50 mm against LABs strains (Lactobacillus sakei ATCC 15521, Lactobacillus plantarum CECT 221 and Carnobacterium piscicola CECT 4020). Against Listeria strains (Listeria innocua NCTC 111288 and Listeria seeligeri NCTC 11289), their antimicrobial activity was better detected in liquid medium assay, evaluating the minimal inhibitory concentration of 50%. BLIS was able to inhibit 60 and 100% of L. seeligeri and L. innocua, respectively, as well as, diluted 1x (v/v) in water was able to inhibit 100% growth of both Listeria. BLIS 17 showed also good results as food preservative when applied in ready-to-eat pork ham artificially contaminated with L. seeligeri in vacuum-package at 4oC during shelf life of 10 days. BLIS was able to maintain low Listeria multiplication, lower samples weight loss, low lipid peroxidation and good color parameters during samples storage. Results demonstrated the importance of optimizing the culture medium to increase microbial mass, to produce and to improve the activity of this antimicrobial molecule. Moreover, results also suggest the possible application of BLIS as a natural food preservative

Bacteriocinas são peptídeos produzidos por várias espécies de bactérias, especialmente bactérias ácido-láticas (BALs) e apresentam um amplo espectro de ação contra bactérias deteriorantes e patógenos de origem alimentar. Entretanto, quando estas bacteriocinas não foram completamente caracterizadas quanto a sequência de seus nucleotídeos e do seu gene correspondente, é recomendada a denominação de substância semelhante a bacteriocina (BLIS). Para aumentar a atividade antimicrobiana de bacteriocinas, a habilidade de BALs probióticas, como Pediococcus pentosaceus, em fermentar diferentes fontes de carbono e nitrogênio tem sido estudado. Para o desenvolvimento de um meio de cultura melhorado, fontes de carbono e nitrogênio devem ser consideradas como nutrientes responsáveis pelo crescimento celular e pela produção de bacteriocina. A melhor condição, após 48 h de cultivo, para o crescimento (3,420 g/L) e para a produção de BLIS por P. pentosaceus ATCC 43200 foi em meio de cultivo Man, Rogosa e Sharp (MRS) suplementado com 1,5% de peptona, pH inicial 6,0 e sob as seguintes condições de cultivo: anaerobiose, 30oC e agitação de 200 rpm. Comparado ao controle (MRS sem suplementação), o crescimento de Pediococcus foi significativamente menor (1,995 g/L) assim, como também, reduziu significativamente o tempo de geração de 2,05 h (controle) para 1,28 h (MRS suplementado), uma redução de aproximadamente 62,5%. Além disso, a adição de peptona ao meio MRS promoveu redução de 4 h para o início da fase exponencial de Pediococcus. Quanto a atividade antimicrobiana de BLIS, a adição de fonte de nitrogênio ao meio MRS também foi bastante significativa. Através do método ágar difusão, BLIS apresentou halos de inibição entre 12,50 a 19,50 mm contra cepas de BALs (Lactobacillus sakei ATCC 15521, Lactobacillus plantarum CECT 221 e Carnobacterium piscicola CECT 4020). Contra cepas de Listeria (Listeria innocua NCTC 11288 e Listeria seeligeri NCTC 11289), a sua atividade inibitória foi melhor detectada em meio líquido, através da determinação da concentração mínima inibitória de 50%. BLIS sem diluição foi capaz de inibir 60 e 100% de L. seeligeri e L. innocua, 15 respectivamente, assim como, diluído 1x (v/v) em água foi capaz de inibir 100% o crescimento de ambas Listeria. BLIS também apresentou bons resultados como conservante de alimento quando aplicado em presunto contaminado artificialmente com L. seeligeri e armazenado a 4oC a vácuo por 10 dias. BLIS foi capaz de manter baixa a multiplicação de Listeria, menor perda de peso das amostras, baixa peroxidação lipídica e bons parâmetros de cor durante o armazenamento das amostras. Os resultados demonstraram a importância de se otimizar meio de cultivo tanto para o aumento da massa microbiana como para a produção e melhoramento da atividade desta molécula antimicrobiana. Além disso, os resultados também sugerem a possível aplicação de BLIS como conservante natural de alimentos

Whole genome sequence of lactic acid bacterium Pediococcus acidilactici strain S1

Abstract Pediococcus acidilactici strain S1, a lactic acid-fermenting bacterium, was isolated from makgeolli-a Korean traditional fermented alcoholic beverage. Here we report the 1,980,172 bp (G + C content, 42%) genome sequence of Pediococcus acidilactici strain S1 with 1,525 protein-coding sequences (CDS), of which 47% could be assigned to recognized functional genes. The genome sequence of the strain S1 might provide insights into the genetic basis of the lactic acid bacterium with alcohol-tolerant.

Draft genome sequence of alcohol-tolerant bacteria Pediococcus acidilactici strain K3

Abstract Pediococcus acidilactici strain K3 is an alcohol-tolerant lactic acid bacterium isolated from nuruk, which is a traditional Korean fermentation starter for makgeolli brewing. Draft genome of this strain was approximately 1,991,399 bp (G+C content, 42.1%) with 1525 protein-coding sequences (CDS), of which 44% were assigned to recognized functional genes. This draft genome sequence data of the strain K3 will provide insights into the genetic basis of its alcohol-tolerance.

Survivability of freeze-dried probiotic Pediococcus pentosaceus strains GS4,GS17 and Lactobacillus gasseri(ATCC 19992)during storage with commonly used pharmaceutical excipients within a period of 120 days

Objective: To examine the survivability and stability of probiotic strains in presence and absence of pharmaceutical excipients for a long period of time at (4 ± 1) ℃. Methods: The survival rates of probiotic strains, Pediococcus pentosaceus GS4 (MTCC12683)(NCBI HM044322),GS17(NCBI KJ608061)and Lactobacillus gasseri (ATCC 19992), were evaluated. Probiotic strains were lyophilized individually and in combination of excipients (sorbitol, ascorbic acid, fructose and skim milk). The prepa-ration was monitored for 120 d storing at(4 ± 1) ℃.During storage,all the preparations were evaluated for viability and stability of probiotic properties like lactic acid produc-tion, antimicrobial effect,water activity,and adherence to epithelial cells. Results: Sorbitol,ascorbic acid and skim milk favoured the viability of freeze-dried cells and sustained probiotic properties during storage. Without excipients (control group), strains showed percentage of survivability not more than 70% while strains with excip-ients survived for 73%–93% for a long period of time. Conclusions: Commonly used excipients can be considered as a vehicle for delivering active principle in probiotic formulation and for sustaining the viability and stability of probiotic strains for a period of 120 d.

Survivability of freeze-dried probiotic Pediococcus pentosaceus strains GS4, GS17 and Lactobacillus gasseri (ATCC 19992) during storage with commonly used pharmaceutical excipients within a period of 120 days

Objective To examine the survivability and stability of probiotic strains in presence and absence of pharmaceutical excipients for a long period of time at (4 ± 1) °C. Methods The survival rates of probiotic strains, Pediococcus pentosaceus GS4 (MTCC12683) (NCBI HM044322), GS17 (NCBI KJ608061) and Lactobacillus gasseri (ATCC 19992), were evaluated. Probiotic strains were lyophilized individually and in combination of excipients (sorbitol, ascorbic acid, fructose and skim milk). The preparation was monitored for 120 d storing at (4 ± 1) °C. During storage, all the preparations were evaluated for viability and stability of probiotic properties like lactic acid production, antimicrobial effect, water activity, and adherence to epithelial cells. Results Sorbitol, ascorbic acid and skim milk favoured the viability of freeze-dried cells and sustained probiotic properties during storage. Without excipients (control group), strains showed percentage of survivability not more than 70% while strains with excipients survived for 73%–93% for a long period of time. Conclusions Commonly used excipients can be considered as a vehicle for delivering active principle in probiotic formulation and for sustaining the viability and stability of probiotic strains for a period of 120 d.

Identification and characterization of Lactobacillus bacterial genera most prevalent used to improve silage digestibility of important forage species for livestock sector.

The objective of this study is to isolate and characterize lactic acid bacteria and their effect as a microbial inoculants in silage digestibility of Lolium perenne –Trifolium pratense (Rye grass - Red clover) (RG-RC), Avena sativa – Vicia sativa (Oat-Vetch) (O-V) and Corn Stover Zea mays (Corn) (C). The lactic acid bacteria (LAB) isolated from three mixtures were identified at 40 day of evolution of micro silages. Morphological, physiological, biochemical and molecular techniques were used to characterize the isolates lactic acid bacteria. The following species were identified from the mixture namely Lactobacillus buchneri, Lactobacillus plantarum, Lactobacillus brevis and Pediococcus acidilactici. 54 micro silages with each feed material were produced, 27 micro silos were inoculated with bacteria’s and the rest was used as a control. The nutritional value of protein, ether extract (EE), ash, energy, neutral detergent fiber (NDF), acid detergent fiber (ADF) at 20, 30 and 40 days of ensilage was compared. The percentage of each in vitro digestibility of treatments performed on day 40 was obtained that corresponded best A-V and RG-TR inoculated with bacteria as they were 35% and 41% more digestible than the control treatments appropriate, concluding that the inoculation of lactic acid bacteria facilitated improved digestibility of silage obtaining good nutritional quality, with optimal values. These results will enable future research on the relationship between LAB species and silage fermentation quality. Use of lactic acid bacteria is recommended as an additive to improve the nutritional quality of food animals as alternative in times of scarcity of fodder or as a supplement to improve the nutritional status of livestock herd.

Molecular identification and evaluation of the probiotic ability of lacticacid bacteria from sow colostrum / Identificación molecular y evaluación de la capacidad probiótica de bacterias ácido lácticas aisladas del calostro de cerdas / Identificação molecular e avaliação da capacidade probiótica de bactérias ácido lácticas isoladas do colostro de porcas

Probiotics have become relevant in animal health, especially because of their ability to replace antibiotic growth promoters. This study assessed the probiotic ability of lactic acid bacteria (LAB) isolated from sow colostrum obtained from pig farms in the southwest of Antioquia, Colombia. Bacteria were identified by molecular methods. Colostrum (1 ml) was collected from 20 sows postpartum. Colostrum (0.1 ml) was plated on Man Rogosa and Sharpe agar (MRS) (Merck), and incubated anaerobically at 37 °C for 48h. Probiotic capacity was determined measuring growth at pH 3,0 and growth on 0.3% w/v ox bile salts. Hemolytic activity, antimicrobial activity against Salmonella typhimurium, and sensitivity to antibiotics commonly used in veterinary were also determined. Molecular identification was made by gen16S Ribosomal analysis method. Of all the strains tested, only BAL 1 and BAL3 showed probiotic ability to grow and withstand an acidic pH (3,0) and 0.3% w/v of ox bile. They had γ-hemolytic activity, were catalase-negative, and inhibited growth of Salmonella typhymurium with halos larger than 11 mm. They were sensitive to amoxicillin, penicillin, chloramphenicol and erythromycin, and were identified as Pediococcus pentosaceus. The existence of Pediococcus pentosaceus in sow colostrum and its probiotic ability was demonstrated.

Los probióticos han cobrado importancia en la salud animal, especialmente por la capacidad de reemplazar los antibióticos promotores de crecimiento. En este trabajo se evaluó la capacidad probiótica de algunas bacterias ácido lácticas (BAL) aisladas de leche calostro de cerdas en granjas del Valle de Aburrá (zona sur); las cuales fueron identificadas por métodos moleculares. Materiales y métodos: Se recolectó 1mL de calostro de 20. Se inocularon 0.1 mL de cada una de las muestras para su aislamiento sobre el agar Man Rogosa Sharpe (MRS) (Merck), incubándolas a 37 °C durante 48h en anaerobiosis. Se determinó la capacidad probiótica realizando las siguiente pruebas: tolerancia a un pH ácido de 3,0, crecimiento en bilis de buey 0,3%P/v, actividad hemolítica, actividad antimicrobiana frente a Salmonella tiphymurium, y sensibilidad a los antibióticos de uso común en veterinaria; y su identificación molecular se realizó por el método análisis del gen16S Ribosomal. De todas las cepas evaluadas solo dos mostraron tener capacidad probiótica al crecer y resistir a un pH ácido de 3,0 y 0,3% P/v de bilis de buey, con actividad γ-hemolitica, catalasa negativa, que inhibieron el crecimiento de Salmonella tiphymurium con halos mayores a 11mm sensibles a Amoxicilina, Penicilina, Cloranfenicol y Eritromicina, e identificadas como Pediococcus pentosaceus. Se demostró la existencia de Pediococcus pentosaceus con características probióticas en el calostro de cerdas.

Os probióticos tem aumentado sua importância na saúde animal, especialmente pela sua capacidade de ser utilizados ao invés dos antibióticos promotores de crescimento. Neste trabalho, avaliou-se a capacidade probiotica de algumas bactérias ácido lácticas (BAL) isoladas de leite de colostro de fêmeas suínas, as amostras foram obtidas de granjas localizadas em Sudeste antioqueño: as bactérias identificaram-se por métodos moleculares. Materiais e métodos: Coletou-se 1ml de colostro em 20 fêmeas suínas recém paridas, das cidades previamente descritas, cultivou-se em superfície 0,1 ml da amostra em ágar Man Rogosa e Sharpe (MRS) (Merck), e foram incubadas a 37 °C/48h em anaerobiose. Determinou-se a capacidade probiótica avaliando o crescimento dos isolados a pH de 3,0 e a sais biliares de boi 0.3%P/v, além, foi determinada a atividade hemolítica, atividade antimicrobiana frente a Salmonella tiphymurium e sensibilidade aos antibióticos de uso comum em veterinária. A identificação molecular realizou-se pelo método de análise do gene 16S Ribosomal. Resultados: de todas as cepas avaliadas só dois, BAL 1 e BAL3 mostraram ter capacidade probiótica ao crescer e resistir a um pH ácido de 3,0 e 0,3% P/v de bílis de boi, com atividade γ-hemolítica, catalase negativa, que inibiram o crescimento de Salmonella tiphymurium com anéis maiores a 11mm, sensíveis a Amoxicilina, Penicilina, Cloranfenicol e Eritromicina, e identificadas como Pediococcus pentosaceus. Conclusão: demostrou-se a existência de Pediococcus pentosaceus com características probióticas no colostro de fêmeas suínas.

Purified dextransucrase from Pediococcus pentosaceus CRAG3 as food additive.

The extracellular crude dextransucrase (0.67 U/mg) from P. pentosaceus CRAG3 (GenBank accession number JX679020) after PEG-1500 fractionation gave specific activity, 20.0 U/mg which by gel filtration resulted in 46.0 U/mg. The purified dextransucrase displayed molecular size of approximately, 224 kDa. The optimum assay conditions for dextransucrase activity were 5% sucrose in 20 mM sodium acetate buffer (pH 5.4) and 30 oC. The dextransucrase was stable up to 40 oC and at pH range of 5.4-7.0. The metal ions such as Co2+, Ca2+, Mg2+ and Zn2+ stimulated the dextransucrase activity by 56, 44, 14 and 12%, respectively. It was most stable at -20 oC with half-life of 307 days. Amongst various additives used, glycerol and Tween 80 provided significant stability to the enzyme with half-life 15.5 and 85.5 h, respectively as compared to control (6.9 h). The solidification of sucrose supplemented milk by purified dextransucrase due to dextran synthesis displayed its application as additive for improving the texture of dairy products.

Evaluation of probiotic properties of Pediococcus acidilactici B14 in association with Lactobacillus acidophilus ATCC 4356 for application in a soy based aerated symbiotic dessert

The aim of this study was to evaluate the probiotic properties of Pediococcus acidilactici B14 and to study its resistance in the gastrointestinal system when combined with Lactobacillus acidophilus ATCC 4356 and used in a potentially symbiotic aerated soy based dessert. P. acidilactici B14 showed some important probiotic characteristics such as survival rate of 45.9% at pH 2.5; 72.4% in 0.3% bile salts and 95.8% after gastrointestinal transit at pH 4.0. Tolerance against the antibiotics cephalexin, neomycin, vancomycin, cefotaxime and penicillin G was also observed. The strain inhibited antagonism against the following cultures: Escherichia coli ATCC 25922, Bacillus cereus ATCC 33018, Staphylococcus aureus ATCC 6538P and Salmonella sp. The mixed culture of P. acidilactici B14 with L. acidophilus ATCC 4356 showed a survival rate of 92.4% after the passage through the gastrointestinal system at pH 4.0. Furthermore, in the presence of the food matrix, an average increase in cell viability, after being subjected to the gastrointestinal system of 9.9% at pH 2.0 and 6.1% at pH 4.0, was observed. This characterized the adequacy of the associated culture as probiotic in the development of a functional food such as soy based aerated symbiotic dessert.

Characterization of Antimicrobial Activities of Pediococcus pentosaceus Vtcc-B-601.

In this study, Pediococcus pentosaceus VTCC-B-601 was investigated and characterized for bacteriocin production. The antimicrobial activities were produced strongly at the late exponential phase (5x108 CFU/ml), corresponding to the activity of cephalosporin (13.3μg/ml) against Salmonella typhimurium ATCC 19430, Pseudomonas aeruginosa ATCC 27853, Staphylococus aureus ATCC 25923, and Micrococcus luteus ATCC 10240. The bacteriocin activity was reduced after proteinase K treatment while the activity was still stable in high temperature. This work supplied a Pediococcus pentosaceus bacteriocin identification that was useful in food preservation, clinical use, and agriculture.

Silagens de milho inoculadas microbiologicamente em diferentes estádios de maturidade: perdas fermentativas, composição bromatológica e digestibilidade in vitro / Corn silage inoculated with lactic acid bacteria in different maturity stages: fermentative losses, chemical composition and in vitro digestibility

Objetivou-se com este estudo avaliar as perdas fermentativas, composição bromatológica e digestibilidade in vitro da matéria orgânica de silagens de milho produzidas em diferentes estádios de maturidade, inoculadas microbiologicamente. Aplicaram-se dois inoculantes comerciais nas silagens produzidas nos estádios SLL, 1/3 LL, 1/2 LL, 2/3 LL e CN, permanecendo ainda um tratamento sem inoculação (silagem controle), configurando um esquema fatorial 3x5. A inoculação com BAL resultou em menores perdas fermentativas (P=0,0348), ao passo que silagens produzidas com plantas mais secas também apresentam menores perdas de MS (P<0,01). A inoculação das silagens resultou em maiores concentrações de PB nas silagens produzidas nos estádios SLL, 2/3 LL e CN (P=0,0033). O uso do inoculante Maize All® resultou em menor concentração de FDN (P=0,0140) no estádio CN e acréscimo dos coeficientes de DIVMO quando as plantas foram colhidas com 2/3 LL e CN (P=0,0006). As perdas fermentativas diminuem devido à utilização dos inoculantes bacterianos e também em silagens produzidas com plantas mais secas. A aplicação de bactérias ácido-láticas (inoculante Maize All®) em silagens de milho produzidas com plantas em estádio de maturidade mais avançado melhora a composição química e digestibilidade in vitro.

The aim of this study was to evaluate the fermentative losses, chemical composition and in vitro organic matter digestibility of corn silages produced in different maturity stages microbiologically inoculated. Two commercial inoculants were applied in the silages produced in stages SLL, 1/3 LL, 1/2 LL, 2/3 LL and CN, remaining an uninoculated treatment (control silage), illustrating a factorial scheme 3 x 5. The lactic acid bacteria (LAB) inoculation resulted in lower fermentative losses (P=0.0348), whereas silages produced with dried plants present lower DM losses (P<0.01). Because of inoculant application, there was higher concentrations of CP in the silages produced in stages SLL, 2/3 LL and CN (P=0.0033) and concentration lower of NDF (P=0.0140) in CN stage because of the Maize All® inoculant. This inoculant provided increase in the IVOMD coefficients when the plants were harvested with 2/3 LL and CN (P= 0.0006). Fermentative losses decreased because of the use of microbial inoculants, and this fact it is also observed in silages produced with dried plants. Application of lactic acid bacteria (Maize All® inoculant) in corn silages produced with plants harvested in more advanced stages of maturity improve the chemical composition and in vitro digestibility.