Identification of microorganisms in wet coffee fermentation Coffea arabica Var Catimor and Castillo in Jardín, Antioquia-Colombia, using culture-dependent methods / Identificación de microorganismos en fermentación húmeda de café Coffea arabica Var Catimor y Castillo en Jardín, Antioquia-Colombia, usando métodos cultivo-dependientes

Background: Mild Colombian coffees are recognized worldwide for their high-quality coffee cup. However, there have been some failures in post-harvest practices, such as coffee grain fermentation. These failures could occasionally lead to defects and inconsistencies in quality products and economic losses for coffee farmers. In Colombia, one of the fermentation methods most used by coffee growers is wet fermentation, conducted by submerging the de-pulped coffee beans for enough time in water tanks to remove the mucilage. Objectives: We evaluated the effect of the water (g)/de-pulped coffee (g) ratio (I: 0/25, II: 10/25, III: 20/25) and final fermentation time (24, 48, and 72 hours) on the total number of microbial groups. We also identified microorganisms of interest as starter cultures. Methods: We used a completely randomized experimental design with two factors; the effect of the water (g)/de-pulped coffee (g) ratio (I: 0/25, II: 10/25, III: 20/25) and final fermentation time (24, 48, and 72 hours), for 9 treatments with two replicates. During the coffee fermentation (1,950 g), the pH and °Brix were monitored. Total counts of different microbial groups (mesophiles, coliforms, lactic-acid bacteria, acetic-acid bacteria, and yeasts) were performed. Various isolates of microorganisms of interest as starter cultures (lactic-acid bacteria and yeasts) were identified using molecular sequencing techniques. Results: 21 lactic-acid bacteria (LAB) isolates and 22 yeasts were obtained from the different mini-batch fermentation systems. The most abundant lactic-acid bacteria species found were Lactiplantibacillus plantarum (46%) and Levilactobacillus brevis (31%). Pichia kluivery (39%) and Torulaspora delbrueckii (22%) were the most abundant yeast species. Conclusion The studied factors did not have effect over the microorganism's development. The identified bacterial and yeasts species have potential as starter cultures for better-quality coffees and in fermentation-related applications.

Antecedentes: Los cafés suaves lavados colombianos son reconocidos a nivel mundial por su buena puntuación sensorial; sin embargo, se han detectado fallas en las prácticas de postcosecha, como lo es la fermentación de los granos de café. Dichas fallas pueden causar defectos y carecer de consistencia en la calidad del producto, ocasionando pérdidas económicas para los caficultores. En Colombia, uno de los métodos más usados por los caficultores es la fermentación húmeda, la cual consiste en sumergir los granos de café despulpado en tanques con agua por un período de tiempo que permita la remoción del mucílago. Objetivos: La presente investigación evaluó la incidencia que tienen la proporción agua/granos despulpados de café (I: 0/25, II: 10/25, III: 20/25) y el tiempo final de fermentación (24, 48 y 72 horas) en el recuento final de grupos microbianos. Por otra parte, se identificaron taxonómicamente microorganismos de interés para su uso como cultivos iniciadores. Métodos: Mini-lotes consistieron en café despulpado (1950 g) puesto en recipientes de plástico abiertos y sumergidos en agua. Se aplicó un diseño experimental completamente aleatorizado de dos factores (proporción agua/ granos de café despulpado y tiempo) a tres niveles, para un total de nueve tratamientos con dos replicas. Durante las fermentaciones de café (1,950 g), el pH y los grados ºBrix, fueron monitoreados. Se realizaron los recuentos totales de los diferentes grupos microbianos: mesófilos, coliformes, bacterias ácido-lácticas, bacterias ácido-acéticas y levaduras. Se identificaron molecularmente diferentes aislados con potencial para ser usados como cultivos iniciadores (bacterias ácido-lácticas y levaduras). Resultados: Los resultados obtenidos mostraron que no hubo diferencia estádisticamente significativa entre los tratamientos aplicados y el recuento final de microorganismos. Un total de 21 aislados de bacterias ácido-lácticas (BAL) y 22 levaduras lograron obtenerse a partir de los diferentes sistemas de fermentación en mini-lote. Las especies de bacterias ácido-lácticas con mayor porcentaje acorde a su identificación taxonómica, corresponden a Lactiplantibacillus plantarum (46%), Levilactobacillus brevis (31%). Las especies de levaduras con mayor porcentaje acorde a su identificación taxonómica corresponden a Pichia kluivery (39%) y Torulaspora delbrueckii (22%). Conclusión Los factores estudiados no afectaron el crecimiento de ninguno de los grupos microbianos presentes en la fermentacion del café. Las especies de microorganismos identificados tienen potencial para se usados como cultivos starter o en aplicaciones dentro de las ciencias de fermentación.

[Agave fructanos promote in vitro biofilm formation with a probiotic consortium Lactobacillus delbrueckii ssp. lactis, L. delbrueckii ssp. bulgaricus and Streptococcus thermophilus]. / Los fructanos de agave promueven la formación de biopelícula in vitro en el consorcio probiótico Lactobacillus delbrueckii ssp. lactis, L. delbrueckii ssp. bulgaricus y Streptococcus thermophilus.

In recent years the relationship between the intestinal microbiota, the host and chronic non-communicable diseases has brought interest into the study of its formation and maintenance in the host. Lactic acid bacteria (BAL) are Gram-positive bacteria with probiotic activity, which have been associated with many health benefits, such as decreased body fat mass and lower risk of type II diabetes mellitus. One of the main colonization mechanisms and bacteria survival strategies is the production of biofilms and the use of prebiotics as substrates to achieve a balance within intestinal microbiota. However, there is not enough evidence to demonstrate the biofilm formation in the presence of agave fructans (AF). This study aimed to evaluate in vitro the biofilm formation in a consortium of lactic acid bacteria: Lactobacillus delbrueckii ssp. lactis, Lactobacillus delbrueckii ssp. bulgaricus y Streptococcus thermophilus in the presence of AF at different concentrations: 0%, 0,1%, 4%, 8% y 16%. The addition of 0,1% of AF correlates with the best capacity for biofilm formation. The findings imply the possibility of modulating the biofilm formation of lactic acid bacteria with AF. These results can contribute positively to the host, by generating intestinal homeostasis, colonization resistance, stability to food digestion and chemical modifications of drugs and carry out beneficial functions to the health.

Silage quality and bacterial diversity of silages inoculated with Listeria monocytogenes and Lacticaseibacillus paracasei_6714

Introduction: King grass (Cenchrus purpureus (Schumach.) Morrone, syn. Pennisetum purpuphoides) and pineapple peel (Ananas comosus) silages are food alternatives for livestock in conditions of feed shortage. Objective: To describe the dynamics of the microbiota present in king grass and pineapple silage during the fermentation process using next generation sequencing (NGS) and to evaluate the protective effect of Lacticaseibacillus paracasei_6714 as a silage inoculum against Listeria monocytogenes. Methods: We used an unrestricted randomized design to characterize the microbiota present in silages made from king grass harvested 70 days after regrowth and pineapple peel. We inoculated mixtures of grass and peel with L. paracasei_6714 or L. monocytogenes, or both, with a non-inoculated treatment as control. The nutritional and fermentative profile was evaluated after 30 days. After 15 and 30 days of fermentation, we used 16S rRNA analysis to determine the dynamics and diversity of the microbiota in the inoculated and control silages. Result: Dry matter content and digestibility did not differ significantly; however, there were differences in crude protein, pH and organic acids. We obtained 4432 amplicon sequence variants of Proteobacteria, Firmicutes, Bacterioidetes, Actinobacteria, Verrucomicrobia, Planctomycetes and Patescibacteria. The relative abundance of each phylum varied depending on the material and fermentation period. Phylum similarity was over 70 % (but not greater than 50 % with Bray-Curtis at the species level). Conclusion: These bacterial communities seem to have an important role during silage fermentation. Proper management of silage processing can reduce or eliminate pathogenic bacteria.

Introducción: Los ensilajes del pasto king grass (Cenchrus purpureus (Schumach.) Morrone, syn. Pennisetum purpuphoides) y cáscaras de piña (Ananas comosus) son alternativas de alimento para ganado en condiciones de escasez alimentaria. Objetivo: Describir las dinámicas de la microbiota presente en los ensilajes de king grass y piña durante el proceso de fermentación usando secuenciación de próxima generación (NGS) y evaluar el efecto de protección de Lacticaseibacillus paracasei_6714 como inoculante de ensilaje ante Listeria monocytogenes. Métodos: Usamos un diseño aleatorio no restringido para caracterizar la microbiota presente en ensilajes de king Grass cosechados 70 días después de rebrote y de cáscaras de piña. Inoculamos mezclas de pasto y cáscara con L. paracasei_6714 o L. monocytogenes, o ambos, con un tratamiento control sin inocular. El perfil nutricional y de fermentación fue evaluado luego de 30 días. Después de 15 y 30 días de fermentación, usamos un análisis de para determinar la dinámicas y diversidad de la microbiota en los ensilajes inoculados y control. Resultados: Los contenidos de materia seca y digestibilidad, no difirieron significativamente; sin embargo, hubo diferencias en proteína cruda, pH y ácidos orgánicos. Obtuvimos 4 432 secuencias variantes de amplicon de Proteobacteria, Firmicutes, Bacterioidetes, Actinobacteria, Verrucomicrobia, Planctomycetes y de Patescibacteria. La abundancia relativa de cada filo vario dependiendo del material y periodo de fermentación. Similitudes de filo fueron mayores al 70 % (pero no mayor que 50 % con Bray-Curtis a nivel de especie). Conclusión: Estas comunidades bacterianas parecen cumplir un papel importante durante la fermentación del ensilaje. Un manejo apropiado del proceso de ensilaje puede reducir o eliminar baterías patogénicas.

Biological control of foodborne pathogens by lacticacid bacteria: A focus on juice processing industries

Abstract The use of lactic acid bacteria (LAB) in foods as biocontrol agents against foodborne pathogens has become increasingly known. Under the premise that controlling the adhesion of microorganisms to food contact surfaces is an essential step for meeting the goals of food processing, the aim of this work was to investigate the inhibitory and anti-biofilm effectiveness of Lactobacillus rhamnosus GG (ATCC 53103) and Lactobacillus casei (ATCC 393) against Escherichia coli O157:H7, Salmonella enterica and Listeria monocytogenes. Lactobacillus strains (108UFCCFU/ml) and pathogens (104UFCCFU/ml) were evaluated to monitor LAB anti-adhesive and antibiofilm effect, in two main scenarios: (i) co-adhesion and (ii) pathogen incorporation to stainless steel surfaces with a protective biofilm of Lactobacillus cells. In (i) the predominant effect was observed in L. rhamnosus against S. enterica and L. monocytogenes, whereas in (ii) both LAB significantly reduced the number of pathogenic adherent cells. The effect of pre-established LAB biofilms was more successful in displacing the three pathogens than when they were evaluated under co-adhesion. These findings show that both LAB can be considered good candidates to prevent or inhibit the adhesion and colonization of L. monocytogenes, S. enterica and E. coli O157:H7 on surfaces and conditions of relevance for juice processing industries, offering alternatives for improving the safety and quality of fruit-based products.

Resumen Existe un creciente interés en el uso de bacterias ácido lácticas (BAL) como agentes de biocontrol frente a patógenos de transmisión alimentaria. Bajo la premisa de que el control de la adhesión de microorganismos a superficies de contacto con alimentos es el paso esencial para evitar su contaminación, el objetivo de este trabajo fue investigar la efectividad inhibitoria y antibiofilm de Lactobacillus rhamnosus GG (ATCC 53103) y Lactobacillus casei (ATCC 393) frente a Escherichia coli O157:H7, Salmonella enterica y Listeria monocytogenes. A fin de cumplir con el objetivo propuesto, las cepas de Lactobacillus (108UFCUFC/ml) y los patógenos (104UFCUFC/ml) se ensayaron en 2 escenarios: (1) coadhesión, y (2) incorporación de los patógenos a las superficies de acero inoxidable con un biofilm preformado de Lactobacillus. En (1), el efecto predominante se observó con L. rhamnosus frente a S. enterica y L. monocytogenes, mientras que en (2), ambas BAL redujeron significativamente el número de células patógenas adheridas. En función de estos resultados, concluimos que el efecto de un biofilm preformado de ambas BAL fue más exitoso en el desplazamiento de los 3 patógenos que en coadhesión. Ambas BAL pueden considerarse buenas candidatas para mitigar la adhesión y colonización de L. monocytogenes, S. enterica y E. coli O157:H7 en superficies en condiciones de relevancia para la industria procesadora de jugos, y, de esta manera, ofrecer alternativas para mejorar la seguridad y calidad de los alimentos a base de frutas.

Calidad microbiológica de embutidos crudos: estudio del caso en Latinoamérica / Microbiology of raw sausages: a case of study in Latin America

Introducción. Los embutidos crudos se componen de carne fragmentada y otros ingredientes no cárnicos (sal, especias, fosfatos, nitritos) pero cuya formulación varía según el país; son productos altamente perecederos y podrían representar un riesgo para el consumidor. Objetivo. Los embutidos frescos son de alto consumo en varios países de Latinoamérica, por ello, el objetivo de esta revisión bibliográfica es compilar la información disponible sobre la calidad microbiológica de este tipo de productos en la región. Materiales y métodos. Se realizó una búsqueda de literatura (desde el 2006 a la fecha) en las principales bases de datos. Resultados. Se determinó que la calidad microbiológica de los embutidos crudos latinoamericanos no es adecuada según la reglamentación. Las bacterias más estudiadas son los microorganismos totales aerobios mesófilos (MTAM), y las bacterias ácido-lácticas (BAL); estos dos grupos son los referentes para determinar la vida útil. Los patógenos más analizados son Salmonella spp. y Listeria monocytogenes y llama la atención que Staphyloccoccus aureus no se utiliza como indicador de malas prácticas de higiene o de inocuidad. Conclusiones. En general se confirma que los embutidos frescos podrían ser un riesgo para la salud pública ya que presentan recuentos microbiológicos altos, en ocasiones no regulados. Algunos agentes antimicrobianos como los compuestos etanólicos de propóleos (EEP), compuestos fenólicos y bacteriófagos han sido estudiados. Sin embargo, no está claro si a nivel artesanal este tipo de ingredientes son utilizados del todo. Finalmente, destaca la necesidad de armonizar las metodologías de estudio y la normativa vigente en los distintos países(AU)

Introduction. Raw sausages are products composed of comminuted meat and other non- meat ingredients (salt, spices, phosphates, nitrites) but the formulation varies in each country. Given this nature, raw sausages are highly perishable and may represent an important risk for consumers. Aim. As raw sausages are highly consumed in many Latin-American countries, the objective of this literature review was to compile the available information about studies of the microbial quality of these products in the region. Materials and methods. For that purpose, a literature search was performed on main data bases to compile studies from 2006 to nowadays. Results. In general, it was found that microbiological quality of Latin-American raw sausages is not adequate according to current regulation. Total aerobic mesophilic microorganisms (TAMM) and Lactic Acid Bacteria (LAB) were the most studied indicators; these two groups are the main reference to establish shelf life. Salmonella spp. and Listeria monocytogenes were the most studied pathogens, and it is noticeable that Staphyloccoccus aureus is not used as an indicator for safety or manipulation. Conclusions. It is perceived that raw sausages in the region could represent a public health risk as they frequently present high microbiological counts, not regulated in many cases. For conservation, antimicrobial agents as propolic ethanoplic extracts (PEE), phenolyc compounds, and bacteriophages have been studied. However, it is not clear if these ingredients are used at the artisanal level, even though it can be assumed that they are not given the high microbial numbers that are reported. Finally, it stands out the need of harmonization of methodologies and current regulation in the countries(AU)

Biological control of foodborne pathogens by lactic acid bacteria: A focus on juice processing industries.

The use of lactic acid bacteria (LAB) in foods as biocontrol agents against foodborne pathogens has become increasingly known. Under the premise that controlling the adhesion of microorganisms to food contact surfaces is an essential step for meeting the goals of food processing, the aim of this work was to investigate the inhibitory and anti-biofilm effectiveness of Lactobacillus rhamnosus GG (ATCC 53103) and Lactobacillus casei (ATCC 393) against Escherichia coli O157:H7, Salmonella enterica and Listeria monocytogenes. Lactobacillus strains (108CFU/ml) and pathogens (104CFU/ml) were evaluated to monitor LAB anti-adhesive and antibiofilm effect, in two main scenarios: (i) co-adhesion and (ii) pathogen incorporation to stainless steel surfaces with a protective biofilm of Lactobacillus cells. In (i) the predominant effect was observed in L. rhamnosus against S. enterica and L. monocytogenes, whereas in (ii) both LAB significantly reduced the number of pathogenic adherent cells. The effect of pre-established LAB biofilms was more successful in displacing the three pathogens than when they were evaluated under co-adhesion. These findings show that both LAB can be considered good candidates to prevent or inhibit the adhesion and colonization of L. monocytogenes, S. enterica and E. coli O157:H7 on surfaces and conditions of relevance for juice processing industries, offering alternatives for improving the safety and quality of fruit-based products.

Cultivo associado de Leuconostoc mesenteroides lb10. 4 e Lactococcus lactis l4a8: propriedades antimicrobianas e potencial aplicação / Associated culture of Leuconostoc mesenteroides lb10. 4 and Lactococcus lactis l4a8: antimicrobial properties and potential application / Cultivo asociado de Leuconostoc mesenteroides lb10. 4 y Lactococcus lactis l4a8: propiedades antimicrobianas y aplicación potencial

O interesse por produtos lácteos funcionais tem motivado o estudo e a prospecção de novas bactérias lácticas. Neste contexto o objetivo deste trabalho foi explorar a potencial associação de Leuconostoc mesenteroides LB10.4 e Lactococcus lactis L4A8 isolados de leite de búfala e identificar aplicações em matriz alimentar. Foram realizados testes de atividade antimicrobiana, influência das bacteriocinas e avaliação da eficiência das bactérias ácido lácticas (BAL) individualmente e associadas frente a espécie de Listeria monocytogenes ATCC 7644 aplicadas em caldo Tryptic Soy Broth e em matriz láctea. Na avaliação da atividade antimicrobiana, Leuconostoc mesenteroides LB 10.4 e Lactococcus lactis L4A8 foram capazes de inibir Staphylococcus aureus ATCC 25923 e Listeria monocytogenes ATCC 7644 com halos de inibição variando de 8 a 16 mm e 6 a 18 mm, respectivamente, pelos dois métodos testados. Na avaliação do efeito das bacteriocinas, os resultados demonstraram melhor controle inibitório dos patógenos pela nisina nas concentrações de 1% e 2%, com halos de inibição entre 14 a 24 mm. A avaliação da eficiência das BAL individualmente e associadas frente a espécie de Listeria monocytogenes ATCC 7644, demonstrou que os isolados em associação são capazes de inibir com mais efetividade a bactéria patogênica, sendo observada uma redução na contagem de L. monocytogenes de 2,67x107 UFC/g para 1,35x104 UFC/g após 240 horas, em matriz alimentar. As bactérias lácticas Leuconostoc mesenteroides LB10.4 e Lactococcus lactis L4A8 apresentaram características promissoras quanto ao seu potencial inibitório, não sendo inibidas pela pediocina. Destaca-se com esses resultados, a importância de estudar o leite de búfala como fonte de novos candidatos de bactérias lácticas autóctones para aplicação em alimentos.(AU)

The interest in functional dairy products has motivated the study and prospection of new lactic bacteria. In this context, the aim of this study was to explore the potential association of Leuconostoc mesenteroides LB10.4 and Lactococcus lactis L4A8 isolated from buffalo milk and to identify applications in food matrix. Tests of antimicrobial activity, influence of bacteriocins and evaluation of the efficiency of lactic acid bacteria (LAB) individually and associated against the species of Listeria monocytogenes ATCC 7644 applied in Tryptic Soy Broth and in milk matrix were performed. In the evaluation of antimicrobial activity, Leuconostoc mesenteroides LB 10.4 and Lactococcus lactis L4A8 were able to inhibit Staphylococcus aureus ATCC 25923 and Listeria monocytogenes ATCC 7644 with inhibition halos ranging from 8 to 16 mm and 6 to 18 mm, respectively, by the two methods tested. In evaluating the effect of bacteriocins, the results showed better inhibitory control of pathogens by nisin at concentrations of 1% and 2%, with inhibition halos between 14 and 24 mm. The evaluation of the efficiency of LAB individually and associated against the species of Listeria monocytogenes ATCC 7644, showed that the isolates in association are able to more effectively inhibit the pathogenic bacteria, with a reduction in the L. monocytogenes count of 2.67x107 CFU /g to 1.35x104 CFU/g after 240 hours, in food matrix. The lactic acid bacteria Leuconostoc mesenteroides LB10.4 and Lactococcus lactis L4A8 showed promising characteristics regarding their inhibitory potential, not being inhibited by pediocin. With these results, attract attention the importance of studying buffalo milk as a source of new candid ates of autochthonous lactic acid bacteria for application in food.(AU)

El interés por los productos lácteos funcionales ha motivado el estudio y prospección de nuevas bacterias ácido lácticas. En este contexto, el objetivo de este trabajo fue explorar la asociación potencial de Leuconostoc mesenteroides LB10.4 y Lactococcus lactis L4A8 aislados de leche de búfala e identificar aplicaciones en matriz alimentaria. Se realizaron pruebas de actividad antimicrobiana, influencia de bacteriocinas y evaluación de la eficiencia de bacterias ácido lácticas (BAL) individualmente y asociadas frente a las especies de Listeria monocytogenes ATCC 7644 aplicadas en caldo Tryptic Soy Broth y en matriz de leche. En la evaluación de la actividad antimicrobiana, Leuconostoc mesenteroides LB10.4 y Lactococcus lactis L4A8 fueron capaces de inhibir Staphylococcus aureus ATCC 25923 y Listeria monocytogenes ATCC 7644 con halos de inhibición de 8 a 16 mm y de 6 a 18 mm, respectivamente, por los dos métodos probados. Al evaluar el efecto de las bacteriocinas, los resultados mostraron un mejor control inhibitorio de patógenos por parte de la nisina a concentraciones de 1% y 2%, con halos de inhibición entre 14 y 24 mm. La evaluación de la eficiencia de las BAL de forma individual y asociada frente a la especie de Listeria monocytogenes ATCC 7644, mostró que los aislados en asociación son capaces de inhibir más eficazmente las bacterias patógenas, con una reducción del recuento de L. monocytogenes de 2,67x107 UFC/g a 1,35x104 UFC/g después de 240 horas, en matriz alimentaria. Las bacterias del ácido láctico Leuconostoc mesenteroides LB10.4 y Lactococcus lactis L4A8 mostraron características promisorias en cuanto a su potencial inhibidor, no siendo inhibidas por la pediocina. Con estos resultados, se destaca la importancia de estudiar la leche de búfala como fuente de nuevos candidatos de bacterias ácido lácticas autóctonas para su aplicación en alimentos.(AU)

Microbial diversity in artisanal cheese produced and commercialized in Vale do Taquari in southern Brazil / Diversidade microbiana encontrada em queijos artesanais produzidos e comercializados no Vale do Taquari no sul do Brasil

Researchers have been utilizing matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to identify bacteria and fungi directly from isolates obtained on culture plates; the resulting mass spectrum is then compared with spectra stored in the instrument software. Hence, a fast analytical response is obtained, and the more spectra are known to compare, the safer and more reliable the interpretation of the method will be. Thus, this study sought to identify the diversity of strains found in 10 samples of artisan cheese produced and commercialized in Vale do Taquari (Rio Grande do Sul State, Brazil) using MALDI-TOF MS. From the analyzed cheese, 22 strains were identified at the species level; one sample presented the pathogen Staphylococcus aureus, two showed the presence of lactic acid bacteria (Lactococcus lactis), and the vast majority (68.18%) of strains were composed of species of the Enterobacteriaceae family, with the prevalence of the genera Escherichia, Enterobacter, and Klebsiella. Escherichia coli was present in 50% of the samples analyzed. This demonstrates the need for greater control during all stages of artisanal cheese production and evaluation of the raw material, including safe practices during milking, so that the product meets the identity and quality parameters suitable for human consumption.

MALDI-TOF MS vendo sendo utilizado em laboratórios para identificar bactérias e fungos diretamente de isolados obtidos em placas de cultura. O espectro de massa resultante é então comparado com espectros armazenados no software do equipamento. Assim, obtém-se uma resposta analítica rápida, sendo que, quanto mais espectros forem conhecidos para comparar, mais seguro e confiável será a interpretação do método. Desta maneira, o presente trabalho teve por objetivo, identificar por MALDI-TOF MS, a diversidade de cepas encontrados em 10 amostras de queijos artesanais produzidos e comercializados no Vale do Taquari, Rio Grande do Sul, Brasil. Dos queijos analisados, 22 cepas foram identificadas a nível de espécie, sendo uma (1) amostra apresentou o patógeno Staphylococcus aureus, duas a presença da bactéria ácido láctica (Lactococcus lactis) e a grande maioria (68,18%) das cepas era composta por espécies da família Enterobacteriaceae, com prevalência dos gêneros Escherichia, Enterobacter e Klebsiella. E. coli estava presente em 50% das amostras analisadas. Isso demonstra a necessidade de um maior controle durante todas as etapas de produção do queijo artesanal, bem como a avaliação da matéria-prima, incluindo práticas seguras durante a ordenha para que o produto atenda aos parâmetros de identidade e qualidade, sendo apto ao consumo humano.

Digestibilidad aparente de materia seca, proteína y carbohidratos de la dieta con ensilado biológico de harina de maíz en Cryphiops (C.) caementarius / Apparent digestibility of dry matter, protein and carbohydrate of the diet with biological corn meal silage in Cryphiops (C.) caementarius

Resumen El objetivo del estudio fue determinar la digestibilidad aparente de la materia seca, proteína y carbohidratos de la dieta con ensilado biológico de harina de maíz en Cryphiops (C.) caementarius. Camarones adultos (4.50 a 7.20 cm de longitud total), se colectaron del río Pativilca (Lima, Perú) y en laboratorio se mantuvieron en recipientes individuales instalados dentro de seis acuarios (cuatro camarones por acuario). La dieta control fue sin ensilado y para la dieta experimental se reemplazó el 75% de la harina de maíz por el ensilado biológico de harina de maíz; ambas dietas tuvieron 1% de óxido crómico como marcador inerte. Los camarones fueron alimentados dos veces por día (40% a las 08:00 h y 60% a las 18:00 h) con el 6% del peso húmedo, durante 10 días. Para la colección de heces, los camarones fueron trasladados a un sistema de caja cribada (12 compartimentos) instalada dentro de un acuario. La digestibilidad aparente se determinó en base seca. Los camarones alimentados con la dieta con ensilado biológico de harina de maíz tuvieron altos coeficientes de digestibilidad aparente de carbohidratos (95.69%), proteína cruda (80.02%) y materia seca (76.19%), que aquellos alimentados con la dieta sin ensilado (p<0.05) que fue de 93.48, 78.21 y 74.36%, respectivamente. Los camarones adultos digieren con eficiencia los carbohidratos de la dieta con ensilado biológico de harina de maíz, en vez de la proteína de dicha dieta.

Abstract The aim of this study was to determine the apparent digestibility of dry matter, protein and carbohydrates of the diet with biological corn meal silage in Cryphiops (C.) caementarius. Adult prawn (4.50 to 7.20 cm total length) was collected from the Pativilca river (Lima, Peru) and in the laboratory each shrimp was kept in individual containers set up in six aquaria (four prawn per aquarium). The experimental diet was supplemented with corn meal biological silage and the control diet was without silage; both diets had 1% chromic oxide. Each diet had three replicates. Prawn was fed twice a day (40% at 08:00 h and 60% at 18:00 h) at 6% of wet weight for 10 days. For feces collection, the prawn was transferred to a system of sieved boxes (12 compartments) installed inside an aquarium. Apparent digestibility was determined on a dry basis. Prawn fed the diet with corn meal biological silage had higher apparent digestibility coefficient for carbohydrates (95.69%), crude protein (80.02%) and dry matter (76.19%) than those fed the diet without silage (p<0.05), which was 93.48, 78.21 and 74.36%, respectively. The adult prawn digested efficiently the carbohydrates of the corn meal biological silage diet, instead of the protein of this diet.

Nisin and -poly-L-lysine as natural antimicrobials towards spoilage-associated Lactobacillus plantarum / Combinação de nisina e -poly-L-lysina como antimicrobianos naturais contra Lactobacillus plantarum associado à deterioração

ABSTRACT: Sausages are highly susceptible to microbial spoilage. Lactic acid bacteria (LAB) is the main group of spoilage bacteria in vacuum packed cooked sausages. To control microbial growth natural antimicrobials have been used as food preservatives. The aim of this study was to identify strains of lactic acid bacteria isolated from spoiled commercial Calabresa sausages and use them in an in vitro challenge with the natural antimicrobials, nisin (NI) and -poly-L-lysine (-PL). Mass spectrometry identification of LAB isolated from sausages using MALDI-TOF revealed a predominance of L. plantarum in the LAB population. RAPD-PCR of L. plantarum strains showed four different genetic profiles. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of NI and -PL, alone and in combination, against a pool of different profiles L. plantarum were determined. MIC of NI and -PL were 0.468 mg/ L and 75 mg/ L; respectively, whereas MBC of NI and -PL were 12.48 mg/L and 150 mg/L, respectively. The combined effect of NI and -PL was determined using concentrations at 1/4 and 1/8 of individual MICs. Synergistic effect was confirmed at both concentrations showing a fractional inhibitory concentration index of 0.5 and 0.2, respectively. The combination of NI and -PL at a small concentration of 0.05 mg/L and 9.375 mg/L, respectively, showed inhibitory effect towards spoilage L. plantarum Results show the potential of the combined use of NI and -PL to control sausage spoilage-associated with lactobacilli.

RESUMO: Linguiças são altamente susceptíveis à deterioração microbiana. As bactérias ácido lácticas (BAL) são o principal grupo de bactérias deteriorantes em linguiças cozidas embaladas a vácuo. Para controlar o crescimento microbiano, antimicrobianos naturais têm sido utilizados como conservantes de alimentos. O objetivo deste estudo foi identificar cepas de bactérias ácido lácticas isoladas de linguiças Calabresa em deterioração e utilizá-las como modelo in vitro com os antimicrobianos naturais nisina (NI) e -poly-L-lysina (-PL). A identificação por espectrometria de massa das BAL isoladas das linguiças usando a técnica de MALDI-TOF revelou predominância de L. plantarum. A PCR-RAPD revelou quatro perfis genéticos diferentes. A concentração inibitória mínima (CIM) e a concentração bactericida mínima (CBM) de NI e -PL, individualmente e em combinação, contra o pool de L. plantarum foram determinadas. O valor de CIM de NI e -PL foi 0,468 mg/L e 75 mg/L, respectivamente, e de CBM de NI e -PL foi 12,48 mg/L e 150 mg/L, respectivamente. O efeito da combinação de NI e -PL foi determinada usando concentrações de 1/4 e 1/8 dos CIMs individuais. O efeito sinérgico foi confirmado em ambas concentrações mostrando um índice de concentração inibitório fracionado de 0,5 e 0,2, respectivamente. A combinação de NI e -PL em baixa concentração, 0,05 mg/L e 9,375 mg/L, respectivamente, mostrou efeito inibitório contra L. plantarum. Os resultados demonstram o potencial do uso combinado de NI e -PL para o controle de lactobacilos associados à deterioração em linguiça.

Nisin and ε-poly-L-lysine as natural antimicrobials towards spoilage-associated Lactobacillus plantarum / Combinação de nisina e ε-poly-L-lysina como antimicrobianos naturais contra Lactobacillus plantarum associado à deterioração

ABSTRACT: Sausages are highly susceptible to microbial spoilage. Lactic acid bacteria (LAB) is the main group of spoilage bacteria in vacuum packed cooked sausages. To control microbial growth natural antimicrobials have been used as food preservatives. The aim of this study was to identify strains of lactic acid bacteria isolated from spoiled commercial Calabresa sausages and use them in an in vitro challenge with the natural antimicrobials, nisin (NI) and ε-poly-L-lysine (ε-PL). Mass spectrometry identification of LAB isolated from sausages using MALDI-TOF revealed a predominance of L. plantarum in the LAB population. RAPD-PCR of L. plantarum strains showed four different genetic profiles. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of NI and ε-PL, alone and in combination, against a pool of different profiles L. plantarum were determined. MIC of NI and ε-PL were 0.468 mg/ L and 75 mg/ L; respectively, whereas MBC of NI and ε-PL were 12.48 mg/L and 150 mg/L, respectively. The combined effect of NI and ε-PL was determined using concentrations at 1/4 and 1/8 of individual MICs. Synergistic effect was confirmed at both concentrations showing a fractional inhibitory concentration index of 0.5 and 0.2, respectively. The combination of NI and ε-PL at a small concentration of 0.05 mg/L and 9.375 mg/L, respectively, showed inhibitory effect towards spoilage L. plantarum Results show the potential of the combined use of NI and ε-PL to control sausage spoilage-associated with lactobacilli.

RESUMO: Linguiças são altamente susceptíveis à deterioração microbiana. As bactérias ácido lácticas (BAL) são o principal grupo de bactérias deteriorantes em linguiças cozidas embaladas a vácuo. Para controlar o crescimento microbiano, antimicrobianos naturais têm sido utilizados como conservantes de alimentos. O objetivo deste estudo foi identificar cepas de bactérias ácido lácticas isoladas de linguiças Calabresa em deterioração e utilizá-las como modelo in vitro com os antimicrobianos naturais nisina (NI) e ε-poly-L-lysina (ε-PL). A identificação por espectrometria de massa das BAL isoladas das linguiças usando a técnica de MALDI-TOF revelou predominância de L. plantarum. A PCR-RAPD revelou quatro perfis genéticos diferentes. A concentração inibitória mínima (CIM) e a concentração bactericida mínima (CBM) de NI e ε-PL, individualmente e em combinação, contra o pool de L. plantarum foram determinadas. O valor de CIM de NI e ε-PL foi 0,468 mg/L e 75 mg/L, respectivamente, e de CBM de NI e ε-PL foi 12,48 mg/L e 150 mg/L, respectivamente. O efeito da combinação de NI e ε-PL foi determinada usando concentrações de 1/4 e 1/8 dos CIMs individuais. O efeito sinérgico foi confirmado em ambas concentrações mostrando um índice de concentração inibitório fracionado de 0,5 e 0,2, respectivamente. A combinação de NI e ε-PL em baixa concentração, 0,05 mg/L e 9,375 mg/L, respectivamente, mostrou efeito inibitório contra L. plantarum. Os resultados demonstram o potencial do uso combinado de NI e ε-PL para o controle de lactobacilos associados à deterioração em linguiça.

[Influence of the diet of Mexican women on the nutritional quality and the presence of beneficial microorganisms in human milk]. / Influencia de la dieta de mujeres mexicanas sobre la calidad nutricional y la presencia de microorganismos benéficos en la leche humana.

INTRODUCTION: Introduction: breast milk is the first fundamental food of newborns and it provides all the sources of energy, nutrients and the immunological protection they need during their first months of life. Unfortunately, there are specific circumstances that imply that the mother cannot feed her baby correctly, since the mother's nutritional needs differ to a certain extent during different periods of life. Especially in breastfeeding, since nutritional needs are increased, due to the loss of nutrients, first by colostrum and then by breast milk. Objective: to demonstrate the influence of the diet of Mexican women on the nutritional quality and the presence of beneficial microorganisms in human milk. Methods: seventy descriptive surveys of nutrition and sampling of milk to women in nursing state were carried out. The milks were subjected to various bromatological and microbiological analyzes to evaluate their nutritional quality and possible probiotic activity. Results: it was shown that the mother's food intake influences the nutritional quality of the milk. Likewise, it affects the development and growth of lactic acid bacteria. Several strains were isolated and identified in human milk of the genus Lactobacillus, as well as pathogenic bacteria such as Lodderomyces elongisporus among others. Conclusions: the mothers' nutrition is directly reflected in the nutritional quality of the milk. It was observed that the amount of essential nutrients of milk such as carbohydrates, lipids and proteins vary according to the diet and life rhythm of the mothers, as well as the significant decrease of lactic bacteria with probiotic potential.

INTRODUCCIÓN: Introducción: la leche materna es el primer alimento fundamental de los neonatos y proporciona todas las fuentes de energía, nutrientes y protección inmunológica que necesitan durante sus primeros meses de vida. Lamentablemente, existen circunstancias específicas que implican que la madre no pueda alimentar correctamente a su bebé, ya que las necesidades nutricionales de la madre difieren en cierta medida durante los diversos periodos de la vida, especialmente en la lactancia, ya que se aumentan las necesidades nutricionales debido a la pérdida de nutrientes, primero por el calostro y luego a través de la leche materna. Objetivo: demostrar la influencia de la dieta de mujeres mexicanas sobre la calidad nutricional y la presencia de microorganismos benéficos en la leche humana. Métodos: se llevaron a cabo 70 encuestas descriptivas de nutrición y toma de muestras de leche a mujeres en estado lactante. Las leches fueron sometidas a diversos análisis bromatológicos y microbiológicos para evaluar su calidad nutricional y posible actividad probiótica. Resultados: se demostró que la ingesta de alimentos de la madre influye en la calidad nutricional de la leche. Asimismo, afecta el desarrollo y crecimiento de las bacterias lácticas. Se aislaron e identificaron diversas cepas en leche humana del género Lactobacillus, además de bacterias patógenas como el caso de Lodderomyces elongisporus, entre otros. Conclusiones: la alimentación de las madres se refleja directamente en la calidad nutricional de la leche. Se observó que la cantidad de nutrientes esenciales de la leche, como son los hidratos de carbono, lípidos y proteínas, varían conforme a la alimentación y el ritmo de vida de las madres, así como la disminución significativa de bacterias lácticas con potencial probiótico.

Ação in vivo de Lactococcus lactis subsp. lactis com potencial probiótico na estabilização de células cancerígenas no epitélio colorretal / In vivo action of Lactococcus lactis subsp. lactis with probiotic potential in the stabilization of cancer cells in the colorectal epithelium

Na última década, a ciência contribuiu significativamente para inúmeros avanços em relação ao tratamento e prevenção do câncer colorretal (CCR), porém, a prevalência global e a taxa de mortalidade permanecem elevadas. Há relatos sobre efeitos benéficos de espécies de Bifidobacterium e Lactobacillus com potencial probiótico na prevenção de CCR. No entanto, a bactéria probiótica Lactococcus lactis subps. lactis é comumente utilizada para fins industriais, não havendo comprovações in vivo sobre seu potencial anticarcinogênico. Visto o interesse emergente dos efeitos benéficos dos probióticos a fim de prevenir ou tratar o CCR, o presente estudo objetivou explorar os efeitos de L. lactis subsp. lactis sobre o CCR. Ratos Wistar receberam doses subcutâneas de 1,2 dimetilhidrazina (DMH) e suspensão de L. lactis subsp. lactis por via oral. Após 20 semanas, os tecidos intestinais foram analisados e de acordo com o resultado, o isolado demonstrou potencial anticarcinogênico contra CCR.(AU)

Ação in vivo de Lactococcus lactis subsp. lactis com potencial probiótico na estabilização de células cancerígenas no epitélio colorretal / In vivo action of Lactococcus lactis subsp. lactis with probiotic potential in the stabilization of cancer cells in the colorectal epithelium

Na última década, a ciência contribuiu significativamente para inúmeros avanços em relação ao tratamento e prevenção do câncer colorretal (CCR), porém, a prevalência global e a taxa de mortalidade permanecem elevadas. Há relatos sobre efeitos benéficos de espécies de Bifidobacterium e Lactobacillus com potencial probiótico na prevenção de CCR. No entanto, a bactéria probiótica Lactococcus lactis subps. lactis é comumente utilizada para fins industriais, não havendo comprovações in vivo sobre seu potencial anticarcinogênico. Visto o interesse emergente dos efeitos benéficos dos probióticos a fim de prevenir ou tratar o CCR, o presente estudo objetivou explorar os efeitos de L. lactis subsp. lactis sobre o CCR. Ratos Wistar receberam doses subcutâneas de 1,2 dimetilhidrazina (DMH) e suspensão de L. lactis subsp. lactis por via oral. Após 20 semanas, os tecidos intestinais foram analisados e de acordo com o resultado, o isolado demonstrou potencial anticarcinogênico contra CCR.

Desenvolvimento de sorvete de kefir com polpa de manga: avaliação sensorial, físico-química e de bactérias ácido láticas / Development of kefir ice cream with mango pulp: sensory evaluation, physico-chemistry and lactic acid bacteria

O kefir é uma bebida probiótica que contribui com benefícios à saúde. A sua utilização em preparações como o sorvete contribui para incorporação de micro-organismos probióticos na formulação podendo estimular seu consumo com objetivo funcional, por meio de uma sobremesa apreciada entre os consumidores. O trabalho objetivou analisar as características sensoriais, físico-químicas e microbiológicas de sorvetes caseiros desenvolvidos com kefir e polpa de manga. A formulação 1 do sorvete foi elaborada com kefir à 5% e a formulação 2 com kefir a 10% (m/v) e avaliadas quanto a aceitação, pH, acidez titulável e contagem de BAL. As análises microbiológicas foram realizadas no leite integral, kefir à 5% e 10 % m/v e nos sorvetes antes do congelamento (T0), após o congelamento final (T29 h) e após uma semana do congelamento (T168 h). O pH do kefir a 5 % diferiu (p < 0,05) daquele encontrado no de 10%. A análise sensorial mostrou que as formulações foram bem aceitas (p > 0,05) em todos os atributos. Após T168 h os sorvetes apresentaram contagem BAL igual a 2,7 x107 para a formulação 1 e 1,1 x108 UFC/mL para a formulação 2. Conclui-se que o kefir apresentou características de um leite fermentado e que o sorvete de kefir com polpa de manga à 5% e 10% apresentaram após T168 h contagem mínima de bactérias ácido lácticas prevista na legislação (AU)

Kefir is a probiotic drink that contributes health benefits. Its use in preparations such as ice cream can stimulate its consumption through a dessert appreciated among consumers, in addition to allowing the ingestion of lactic acid bacteria. The objective of this work was to analyze the sensorial, physicalchemistry and microbiological characteristics of homemade ice creams developed with kefir and mango pulp. Formulation 1 of the ice cream was made with kefir at 5% and formulation 2 with kefir at 10%, (w/v) and was evaluated for acceptance, pH, titratable acidity and lactic acid bacteria count. Microbiological analyzes were performed in whole milk, kefir at 5% and 10%, and in ice cream before freezing (T0), after the final freezing (T29 h) and after one week of freezing (T168 h). The pH of the kefir at 5% differed (p <0.05) from that found in 10%. Sensory analysis showed that the formulations were accepted (P> 0.05) in all attributes. After T168 h, the ice creams had a lactic acid bacteria count (AU)

Effect of protein source and probiotic on the intestinal tract of pacific white shrimp Litopenaeus vannamei / Efeito da fonte de proteína e probiotico sobre trato intestinal do camarão branco pacífico Litopenaeus vannamei

This study aimed to evaluate the viability of supplementing two diets for the shrimp Litopenaeus vannamei with Lactobacillus plantarum. One using fish meal as a protein source and another using soy protein concentrate, as well as the effect of these formulations on shrimp intestinal microbiota. To assay probiotic viability in the formulated diets, the number of CFU g-1 was observed weekly over the course of four weeks. The viability of Lactobacillus plantarum in relation to the physical parameters of the diets, including stability, buoyancy, and expansion, was quantified. The effect of the diets on microbiota and intestinal tract morphology was determined by performing a 2x2 factorial experiment (two diets, with or without supplementation) in triplicate, totaling 12 experimental units, with five animals per unit, fed with 3.5% of biomass for 17 days. The concentration of lactic acid bacteria decreased over time, irrespective of protein source. The diet with fishmeal as a protein source, independent of probiotic supplementation, presented good stability and did not disintegrate after four hours. In contrast, the diet with soy protein concentrate, supplemented or not, disintegrated between 2.5 and 3 hours, presenting low stability. All diets presented 0% buoyancy. The expansion rate was higher in diets with soy protein concentrate, but without the influence probiotic supplementation or interaction between the factors. In the in vivo assay, both supplemented diets showed greater total heterotrophic bacteria count than without probiotic; however, no difference in count was noted in diets with different protein source. Lactic acid bacteria were only observed in the shrimp fed diets supplemented with probiotic. Histology of the intestinal tract showed that all intestines had intact, well-developed and well-organized cells, irrespective of diet. Thus, L. plantarum, when combined with different protein sources, produced similar effects on...

Este trabalho objetivou avaliar viabilidade da suplementação de duas dietas ao camarão Litopenaeus vannamei com Lactobacillus plantarum. Uma usando farinha de peixe como fonte proteica e outra usando concentrado de proteico de soja, bem como o efeito destas formulações na microbiota intestinal de camarão. Para testar a viabilidade probiótica nas dietas, o número de UFC g-1 foi observado semanalmente ao longo de quatro semanas. A viabilidade de Lactobacillus plantarum em relação aos parâmetros físicos das dietas, incluindo estabilidade, flutuabilidade e expansão, foram testados. O efeito das dietas na microbiota e na morfologia do trato intestinal foi determinado por meio de um experimento fatorial 2x2 (duas dietas, com ou sem suplementação), em triplicata, totalizando 12 unidades experimentais, com cinco animais por unidade, alimentados com 3,5% de biomassa por 17 dias. A concentração de bactérias do ácido láctico diminuiu ao longo do tempo, independentemente da fonte de proteína. A dieta com farinha de peixe como fonte proteica, independente da suplementação probiótica, apresentou boa estabilidade e não se desintegrou após quatro horas. Em contrapartida, dieta com concentrado proteico de soja, suplementado ou não, desintegrou-se entre 2,5 e 3 horas, apresentando baixa estabilidade. Todas as dietas apresentaram flutuabilidade de 0%. A taxa de expansão foi maior nas dietas com concentrado proteico de soja, mas sem influência da suplementação probiótica ou interação entre os fatores. No ensaio in vivo, ambas as dietas suplementadas apresentaram maior contagem de bactérias heterotróficas totais; no entanto, nenhuma diferença na contagem foi observada em dietas com diferentes fontes de proteína. Bactérias ácido-lácticas só foram observadas em dietas suplementadas com probiótico. A histologia do trato intestinal mostrou que todos intestinos tinham células intactas, bem desenvolvidas e organizadas, independentemente da dieta. Assim, L. plantarum, quando...

Effect of protein source and probiotic on the intestinal tract of pacific white shrimp Litopenaeus vannamei / Efeito da fonte de proteína e probiotico sobre trato intestinal do camarão branco pacífico Litopenaeus vannamei

This study aimed to evaluate the viability of supplementing two diets for the shrimp Litopenaeus vannamei with Lactobacillus plantarum. One using fish meal as a protein source and another using soy protein concentrate, as well as the effect of these formulations on shrimp intestinal microbiota. To assay probiotic viability in the formulated diets, the number of CFU g-1 was observed weekly over the course of four weeks. The viability of Lactobacillus plantarum in relation to the physical parameters of the diets, including stability, buoyancy, and expansion, was quantified. The effect of the diets on microbiota and intestinal tract morphology was determined by performing a 2x2 factorial experiment (two diets, with or without supplementation) in triplicate, totaling 12 experimental units, with five animals per unit, fed with 3.5% of biomass for 17 days. The concentration of lactic acid bacteria decreased over time, irrespective of protein source. The diet with fishmeal as a protein source, independent of probiotic supplementation, presented good stability and did not disintegrate after four hours. In contrast, the diet with soy protein concentrate, supplemented or not, disintegrated between 2.5 and 3 hours, presenting low stability. All diets presented 0% buoyancy. The expansion rate was higher in diets with soy protein concentrate, but without the influence probiotic supplementation or interaction between the factors. In the in vivo assay, both supplemented diets showed greater total heterotrophic bacteria count than without probiotic; however, no difference in count was noted in diets with different protein source. Lactic acid bacteria were only observed in the shrimp fed diets supplemented with probiotic. Histology of the intestinal tract showed that all intestines had intact, well-developed and well-organized cells, irrespective of diet. Thus, L. plantarum, when combined with different protein sources, produced similar effects on...(AU)

Este trabalho objetivou avaliar viabilidade da suplementação de duas dietas ao camarão Litopenaeus vannamei com Lactobacillus plantarum. Uma usando farinha de peixe como fonte proteica e outra usando concentrado de proteico de soja, bem como o efeito destas formulações na microbiota intestinal de camarão. Para testar a viabilidade probiótica nas dietas, o número de UFC g-1 foi observado semanalmente ao longo de quatro semanas. A viabilidade de Lactobacillus plantarum em relação aos parâmetros físicos das dietas, incluindo estabilidade, flutuabilidade e expansão, foram testados. O efeito das dietas na microbiota e na morfologia do trato intestinal foi determinado por meio de um experimento fatorial 2x2 (duas dietas, com ou sem suplementação), em triplicata, totalizando 12 unidades experimentais, com cinco animais por unidade, alimentados com 3,5% de biomassa por 17 dias. A concentração de bactérias do ácido láctico diminuiu ao longo do tempo, independentemente da fonte de proteína. A dieta com farinha de peixe como fonte proteica, independente da suplementação probiótica, apresentou boa estabilidade e não se desintegrou após quatro horas. Em contrapartida, dieta com concentrado proteico de soja, suplementado ou não, desintegrou-se entre 2,5 e 3 horas, apresentando baixa estabilidade. Todas as dietas apresentaram flutuabilidade de 0%. A taxa de expansão foi maior nas dietas com concentrado proteico de soja, mas sem influência da suplementação probiótica ou interação entre os fatores. No ensaio in vivo, ambas as dietas suplementadas apresentaram maior contagem de bactérias heterotróficas totais; no entanto, nenhuma diferença na contagem foi observada em dietas com diferentes fontes de proteína. Bactérias ácido-lácticas só foram observadas em dietas suplementadas com probiótico. A histologia do trato intestinal mostrou que todos intestinos tinham células intactas, bem desenvolvidas e organizadas, independentemente da dieta. Assim, L. plantarum, quando...(AU)

Development and storage studies of high density macrocapsules containing Lactobacillus spp. strains as nutritional supplement in young calves / Desarrollo de macrocápsulas con cepas de Lactobacillus spp. en alta densidad como suplemento nutricional de terneros jóvenes y análisis de su viabilidad durante el almacenamiento

The aim of this study was to evaluate different production methodologies of probiotic macrocapsules with high bacterial densities destined to lactating calves. Three types of capsules containing Lactobacillus casei DSPV318T and Lactobacillus plantarum DSPV354T were prepared from an overnight culture in whey medium: (1) mixing the culture with calcium alginate and then, reincubating the capsules in whey (RC); (2) concentrating the biomass by centrifugation and mixing the pellet with calcium alginate (CC) at different concentrations with respect to the initial culture (5X and 12.5X); (3) CC with cryoprotectants: whey permeate (Per) and glycerol (Gly). Chitosan coating was evaluated. Capsules were freeze-dried and viability was assessed before freezing, after freeze-drying and every two weeks for 84 days of storage at room temperature, 4°C and-20°C. CC showed higher cell densities than RC. Storage temperature affected viability: greater viability at lower temperature. Moreover, the effect of temperature was influenced by other factors, such as capsule coating, culture neutralization and cryoprotectants. Coating improved viability at room temperature; however no effect was observed at 4°C. Culture neutralization allowed greater survival during storage. Cryoprotectants improved viability during freezing, but they also generated a positive or negative effect depending on storage temperature. The best results were: at refrigeration Gly12.5X exhibited counts above 10(9) CFU/capsule until day 70 and Per12.5X until day 56 of storage and at-20°C Gly12.5X showed counts above 10(9) CFU/capsule until the end of the study (84 days). A 10(9) CFU capsule is the daily dose per calf which would facilitate the administration of this probiotic inoculum to field animals.

El objetivo de este estudio fue evaluar diferentes metodologías de producción de macrocápsulas probióticas con altas densidades bacterianas, destinadas a terneros lactantes. Se prepararon cápsulas con Lactobacillus casei DSPV318T y L. plantarum DSPV354T a partir de cultivos overnight en suero de queso, de 3 maneras: 1) mezclando el cultivo con alginato de calcio y luego reincubando las cápsulas en suero (RC); 2) concentrando la biomasa por centrifugación y mezclando el sedimento con alginato de calcio (CC) en diferentes concentraciones con respecto al cultivo inicial (5X y 12,5X), y 3) CC con crioprotectores: permeado de suero (Per) o glicerol (Gly). Se evaluó el recubrimiento con quitosano. Las cápsulas se liofilizaron y se evaluó la viabilidad antes de la congelación, después de la liofilización y cada 2 semanas durante 84 días de almacenamiento a temperatura ambiente a 4 o a-20°C. Con el sistema CC se alcanzaron mayores densidades celulares que con el RC. La temperatura de almacenamiento afectó la viabilidad: se observó una mayor viabilidad a menor temperatura. Además, otros factores incidieron en el efecto de la temperatura: el recubrimiento de las cápsulas, la neutralización del cultivo y la adición de crioprotectores. El recubrimiento mejoró la viabilidad a temperatura ambiente, pero no se observó ningún efecto a 4°C. La neutralización del cultivo permitió una mayor supervivencia durante el almacenamiento. Los crioprotectores mejoraron la viabilidad durante la congelación, pero también generaron un efecto positivo o negativo dependiendo de la temperatura de almacenamiento. Con el almacenamiento en refrigeración se obtuvieron recuentos por encima de 10(9) UFC/cápsula hasta el día 70 y con Gly12,5X y hasta el día 56 con Per12,5X. Con el almacenamiento a-20 °C se obtuvieron cápsulas Gly12,5X con más de 10(9) UFC/cápsula hasta el final del estudio (84 días). Una cápsula de 10(9) UFC es la dosis diaria por ternero que facilitaría la administración de este inóculo probiótico a los animales en el campo.

Development and storage studies of high density macrocapsules containing Lactobacillus spp. strains as nutritional supplement in young calves.

The aim of this study was to evaluate different production methodologies of probiotic macrocapsules with high bacterial densities destined to lactating calves. Three types of capsules containing Lactobacillus casei DSPV318T and Lactobacillus plantarum DSPV354T were prepared from an overnight culture in whey medium: (1) mixing the culture with calcium alginate and then, reincubating the capsules in whey (RC); (2) concentrating the biomass by centrifugation and mixing the pellet with calcium alginate (CC) at different concentrations with respect to the initial culture (5X and 12.5X); (3) CC with cryoprotectants: whey permeate (Per) and glycerol (Gly). Chitosan coating was evaluated. Capsules were freeze-dried and viability was assessed before freezing, after freeze-drying and every two weeks for 84 days of storage at room temperature, 4°C and -20°C. CC showed higher cell densities than RC. Storage temperature affected viability: greater viability at lower temperature. Moreover, the effect of temperature was influenced by other factors, such as capsule coating, culture neutralization and cryoprotectants. Coating improved viability at room temperature; however no effect was observed at 4°C. Culture neutralization allowed greater survival during storage. Cryoprotectants improved viability during freezing, but they also generated a positive or negative effect depending on storage temperature. The best results were: at refrigeration Gly12.5X exhibited counts above 109CFU/capsule until day 70 and Per12.5X until day 56 of storage and at -20°C Gly12.5X showed counts above 109CFU/capsule until the end of the study (84 days). A 109CFU capsule is the daily dose per calf which would facilitate the administration of this probiotic inoculum to field animals.

Control del deterioro microbiológico de filetes de tilapia mediante la aplicación de bacterias lácticas / Control Microbiological Deterioration of Tilapia Fillets by the Application of Lactic Acid Bacteria / Controle de Deterioração Microbiológica da Tilápia Arquiva Através da Aplicação de Bactérias Láticas

Resumen En los últimos años se ha reconocido el potencial de las bacterias ácido lácticas (BAL) como agentes bioconservantes de productos cárnicos, incluido el pescado, al reducir el deterioro microbiológico y químico. El objetivo de este trabajo fue determinar el efecto bioconservante de dos cepas de BAL y el tiempo de impregnación en la calidad microbiológica y química de filetes de tilapia. Se evaluaron dos tiempos de impregnación (1h y 2h) y las BAL Lactobacillus plantarum JCM 1149 y Lactobacillus acidophilus ATCC4356. Los filetes bioconservados (FB) se almacenaron a 5°C y se evaluaron los cambios en el recuento de BAL, mesófilos, psicrófilos y coliformes totales durante 30 días. Los recuentos en los FB fueron de 5.94 de Log UFC/g. de BAL, y <2.7 log UFC/g de coliformes y psicrófilos. Los filetes control presentaron recuentos de 1.2 log UFC/g de BAL mientras los coliformes totales y psicrófilos superaron el límite permitido para consumo humano luego de 10 días. Luego de 10 días. El tratamiento que mantuvo la calidad microbiológica y química de los FB por más tiempo fue L. plantarum con inmersión de una hora. Las BAL estudiadas mostraron un efecto inhibitorio de la microflora deteriorante del pescado y una reducción en la formación de nitrógeno volátil, siendo un método viable para la conservación del pescado.

Abstract In recent years, it has recognized the potential of lactic acid bacteria (LAB) as biopreservatives agents for meat products, including fish, by reducing the microbiological and chemical spoilage. The aim of this study was to determine the effect of two strains biopreservative LAB and impregnation time on the microbiological and chemical quality of tilapia fillets. Two impregnation time (1h and 2h) and two LAB (Lactobacillus plantarum JCM 1149 and Lactobacillus acidophilus ATCC4356 were evaluated. The biopreserved fillets (BF) were stored at 5 ° C and the changes of the LAB count, mesophilic, psychrophilic and total coliforms., were evaluated for 30 days. The microbiological analysis show for the BF 5.94 log CFU / g. of LAB and <2.7 log CFU / g of coliform and psychrophilic bacteria. The control Fillets showed 1.2 log CFU / g of LAB while the psychrophile and total coliforms exceeded the allowed limit for human consumption after 10 days. The treatment kept the microbiological and chemical quality of the BF longer was L. plantarum with one hour of impregnation. The LAB studied showed an inhibitory effect of the spoilage microflora of fish and a reduction in the formation of volatile nitrogen compounds, being a viable fish preservation method.

Resumo Nos últimos anos tem se reconhecido o potencial das bactérias ácido lácticas (BAL) como agentes bioconservantes de pro dutos carnicos, incluído o peixe ao diminuir o deterioro microbiológico e químico. O propósito deste estudo foi determinar o efeito bioconservante de duas cepas de BAL e a avaliação do tempo de impregnação requerido para obter uma boa qua lidade microbiológica e química de filetes de tilapia. Foram avaliados dois tempos de impregnação (1h e 2h) com as BAL Lactobacillus plantarum JMC 1149 e Lactobacillus acidophilus ATCC4356. Os filetes biopreservados (FB) foram estocados a 5oC e valoradas as modificações na contagem de microorganismos BAL, mesófilos, psicrófilos, coliformes totais durante 30 dias. As contagens microbianas nos FB foram de 5.94 de log UFC/g e <2.7 log UFC/g de coliformes e psicrofilos. Os filetes controle apresentaram valores de 1.2 log UFC/g de BAL, não entanto os coliformes totais e psicrófilos superaram o limite permitido para consumo humano após 10 dias de estocagem. O tratamento que manteve a qualidade microbiológica e química dos FB por maior tempo foi de imersão com Lactobacillus plantarum por uma hora. As BAL estudadas amostraram um efeito inibitório da microflora deteriorante dos peixes e uma redução na formação de nitrogênio volátil se apresentan do como um método viável para a preservação de filete de peixe