Abstract The aim of this study was to evaluate the performance of Caspian Sea-style spontaneous milk fermentation to improve the quality of pasteurized milk containing high levels of Pseudomonas contamination, with a focus on microbiological safety and stability of the final product. Bacterial diversity of pasteurized milk, fermentation process, and after 60 days of storage was analyzed by Illumina-based sequencing, and presence of viable taxa was confirmed by culturing on selective media. Low quality pasteurized milk harbored mainly Gram-negative bacteria, markedly dominated by Pseudomonas. Following fermentation, lactic acid bacteria rapidly became dominant with maximum population of 10.15 log CFU/mL at 18 h, represented mainly by Lactococcus. However, sequences related to Pseudomonas, and to a lesser extent for enterobacteria, remained constant throughout the fermentation process. The cultured-dependent approach confirmed the presence of viable Pseudomonas, with a final population of 5.60 log CFU/mL. Biochemical transformations were further analyzed, indicating lactic acid as the main end-metabolite produced (maximum concentration of 5.93 g/L at 24 h). In addition, the increase of 2-nonanone can be correlated as a volatile biomarker of P. aeruginosa and related species. Altogether, the results demonstrated that natural milk fermentation may often not inhibit the development of pathogens and food spoilage microorganisms.

Abstract Food supplements have been increasingly investigated. Probiotics have several benefits for human and animal health and selenium (Se) is widely recommended against oxidative stress. In this context, the aim of this study was to develop a low-cost bioprocess to produce a functional food product comprising both probiotic and Se accumulation. Yeast cells of Saccharomyces boulardii CCT 4308 were cultivated using sugarcane molasses as substrate. Optimization studies were performed to evaluate the best medium composition for biomass production and Se-accumulation in batch and fed-batch systems. Optimized conditions were defined with a medium composed of 150 g L-1 sugarcane molasses and 12 g L-1 yeast extract, with feeding of 100 g L-1 sugarcane molasses and 100 μg mL-1 of Se incorporation after 4 h and 10 h of fermentation, respectively, during 48 h in STR (stirred tank reactor). Best biomass production reached 14.52 g L-1 with 3.20 mg Se g-1 biomass at 12 h. Process optimization led to 4.82-fold increase in biomass production compared to initial condition. A final Se-enriched S. boulardii CCT 4308 biomass was obtained, which is comparable to commercial products. An alternative probiotic yeast biomass was efficiently produced as a new food-form of Se supplement in a sustainable process using an inexpensive agro-industrial residue.

RESUMEN El objetivo de este estudio fue evaluar varios métodos de ruptura de las esporas de Ganoderma lucidum y extraer sus propiedades bioactivas. Para este propósito se evaluaron diferentes técnicas de rompimiento como: la maceración manual (RM), la maceración con esferas de diversos materiales (BR) y la exposición a microondas junto la maceración de las esporas con esferas de acero/cromo (MBR1). La ruptura de las esporas fue evaluada por espectroscopia UV-Vis, la cual mostró que las vibraciones 2955, 1642, 1240, 1080 y 1746 cm-1 correspondieron a cambios estructurales en las paredes de las esporas. El extracto MBR1 presento el mayor contenido de carbohidratos (19,80 mg.g-1) y polifenoles (2,21 mg.g-1), mientras que el extracto BR tuvo una mayor actividad antioxidante (57,22% Inb DPPH). Los extractos MBR1 y BR también presentaron en el análisis de monosacáridos un 62,2 y 73,5% de contenido glucosa. Como conclusión la mejor metodología para extraer biomoléculas de las paredes de las esporas de G. lucidum fueron el tratamiento térmico con microondas y la ruptura de las paredes con esferas de acero/cromo, porque este proceso permitió la extracción de una mayor cantidad de carbohidratos con posibles propiedades antioxidantes.

SUMMARY This study aimed at evaluating effective methods for breaking the hard and insoluble spores of Ganoderma lucidum to recover functional biomolecules. Rupture techniques were evaluated such as manual maceration (RM), maceration with spheres of various materials (BR), and microwave exposure plus maceration with steel/ chrome spheres (MBR1). Spore rupture was evaluated using UV-Vis spectroscopy, which showed vibrations of 2955, 1642, 1240, 1080 and 1746 cm-1 corresponding to changes in spore walls. The MBR1 extract contained the largest amounts of carbohydrates (19.80 mg.g-1 spores) and polyphenols (2.21 mg.g-1 spores), whereas the BR extract had higher antioxidant activity (57.22%Inb DPPH). The MBR1 and BR extracts contained 62.2 and 73.5% glucose, respectively. Both methods also involved significant extraction of carbohydrates and proteins. The best way to extract biomolecules from spore walls is to perform a microwave heat treatment and break the walls with steel/chrome spheres; this produces large quantities of carbohydrates with antioxidant properties.

The objective of this work was to evaluate the performance of piglets in the nursery phase submitted to activation of immune system with vaccine against Haemophilus parasuis and to different levels of metabolizable energy in the diets. It was used 160 barrows and females at 30 days of age, weaned at 23 days of age, with initial weight of 7.97 ± 0.54 kg, distributed in 2 × 4 factorial arrangement (with vaccination and without vaccination with four levels of metabolizable energy - 3,200; 3,300; 3,400 and 3,500 kcal/kg). Each combination was evaluated with five repetitions of four animals per experimental plot (two males and two females). Energy levels reduced linearly the intake, feed conversion and the total number of leukocytes in blood in the experimental period, regardless of the use of vaccine which in turn reduced daily average weight gain only in the first week after second application and it increased the number of limonocytes and spleen weight after three weeks. There was no difference in the C-reactive protein quantity in the blood in the periods when it was evaluated. The use of vaccine reduces performance immediatelly after its application but after it occurs recovery of performance with changes in the number of monocytes and spleen weight. Dietary energy levels do not smooth this reduction in the performance, but they influence intake, feed conversion and blood parameters in the total period of use.

Microbial ecology and chemical composition of Brazilian kefir beverage was performed. The microorganisms associated with Brazilian kefir were investigated using a combination of phenotypic and genotypic methods. A total of 359 microbial isolates were identified. Lactic acid bacteria (60.5%) were the major isolated group identified, followed by yeasts (30.6%) and acetic acid bacteria (8.9%). Lactobacillus paracasei (89 isolates), Lactobacillus parabuchneri (41 isolates), Lactobacillus casei (32 isolates), Lactobacillus kefiri (31 isolates), Lactococcus lactis (24 isolates), Acetobacter lovaniensis (32 isolates), Kluyveromyces lactis (31 isolates), Kazachstania aerobia (23 isolates), Saccharomyces cerevisiae (41 isolates) and Lachancea meyersii (15 isolates) were the microbial species isolated. Scanning electron microscopy showed that the microbiota was dominated by bacilli (short and curved long) cells growing in close association with lemon-shaped yeasts cells. During the 24 h of fermentation, the protein content increased, while lactose and fat content decreased. The concentration of lactic acid ranged from 1.4 to 17.4 mg/ml, and that of acetic acid increased from 2.1 to 2.73 mg/ml. The production of ethanol was limited, reaching a final mean value of 0.5 mg/ml.

O objetivo neste trabalho foi avaliar o uso de aditivos à base de extratos vegetais associados ou não ao uso de simbióticos como alternativa aos antibióticos como promotores de crescimento para suínos na fase inicial. Foram utilizados 125 leitões de genética comercial, com peso inicial de 7,4 ± 1,1 kg, desmamados aos 20 dias, durante 50 dias. O delineamento experimental foi o de blocos casualizados (peso inicial) com cinco tratamentos e cinco repetições com cinco animais por unidade experimental. As dietas experimentais foram uma ração basal, sem aditivo, e outras quatro, uma contendo antibiótico (colistina + óxido de zinco); outra com 0,3% de uma mistura simbiótica; outra com 0,3% da mistura simbiótica e 0,05% de uma mistura herbal (extrato de plantas e óleos essenciais microencapsulados) e outra com 0,3% da mistura simbiótica e 0,10% da mistura herbal. Foi observado que o uso da mistura simbiótica associada a 0,05% da mistura herbal teve resultados positivos e semelhantes aos obtidos com as dietas contendo antibióticos quanto ao peso final, ganho de peso, consumo médio diário, pH do estômago e incidência de diarreia. O uso de simbióticos promoveu menor relação vilosidade:cripta, exceto quando associado a 0,1% da mistura herbal, que se assemelhou à dieta sem aditivos. Não houve diferenças na conversão alimentar, na contagem de coliformes e lactobacilus no jejuno e no ceco e no pH do ceco. O uso de uma mistura herbal associada a simbióticos é uma alternativa viável para substituição ao uso de antibióticos como promotores de crescimento para suínos na fase inicial. No entanto, a concentração da mistura herbal é um fator importante a ser considerado.

The objective of this work was to evaluate the use of vegetable extract based addictives associated or not to symbiotics as an alternative for antibiotic as growth promoters for piglets at initial phase. It was used 125 pigs of commercial genetic line with initial weight of 7.4 ± 1.1 kg, weaned at 20 days of age, during 50 days. The experimental design used was randomized blocks (initial weight), with five treatments and five replicates with five animals per experimental unit. The experimental diets were a basal diet, without additive, and other four diets, one diet containing antibiotic (colistin + zinc oxide); diet with 0.3% of a symbiotic mixture; diet with 0.3% of a symbiotic mixture plus 0.05% of herbal mix (plant extracts and microencapsulated essential oils); and another diet with 0.3% of symbiotic mixture plus 0.1% of herbal mixture. It was observed that the use of the symbiotic mixture associated to 0.05% of herbal mixture showed positive results, similar to the ones obtained with diets containing antibiotics for the final weight, average weight gain, average daily feed intake and pH of stomach and diarrhea incidence. The use of symbiotics promoted smaller vilous:depth crypt ratio, except when associated to 0.1% of herbal mixture, which was similar to the diet without addictives. There was no differences in the feed conversion, coliform and lactobacillus count in the jejune and in the cecum and in the pH of cecum. The use of herbal mixture associated to symbiotic is a viable alternative for replacing the use of antibiotics as growth promoters for piglets in the initial phase. However, the concentration of the herbal mixture is an important factor to be considered.