Elongated cells of Listeria monocytogenes in biofilms in the presence of sucrose and bacteriocin-producing Leuconostoc mesenteroides A11

Listeria monocytogenes is a foodborne pathogen which may survive in biofilms and persist in food processing plants. In this study, the ability of Leuconostoc mesenteroides (bac+ and bac-) to inhibit biofilm formation by L. monocytogenes ATCC 19115 was studied with stainless steel coupons immersed in BHI broth and BHI broth plus sucrose in combination with the Lactic Acid Bacteria (LAB). Adhered cells were collected with swabs and enumerated on selective agars (Oxford for listeria and MRS for leuconostoc). Leuconostoc mesenteroides bac+ in co-culture with L. monocytogenes was effective to inhibit biofilm formation by listeria for up to 3 hours of incubation, but at 24 hours, biofilm was present in all conditions tested, as confirmed by observations of stainless steel coupons under Scanning Electron Microscopy (SEM). It was also observed that in the presence of L. mesenteroides bac+ in BHI plus sucrose, a high number of elongated cells of L. monocytogenes was present, which may indicate an adaptation response of the pathogen to stress conditions with important implications for food safety.

Partial purification and characterization of a bacteriocin produced by Enterococcus faecium 130 isolated from mozzarella cheese

Lactic acid bacteria are important in foods as potential probiotics and also due to the ability to produce antimicrobial compounds that can contribute for biopreservation. In this work, the bacteriocin produced by the food isolate Enterococcus faecium 130 was partially purified and characterized. The compound was active against Gram-positive bacteria, including Listeria monocytogenes. It was produced after 4 days of storage at a broad temperature range (4 to 37 °C); it was stable at pH ranging from 2 to 10 with no loss of activity after heating at 100 °C for 15 minutes. Bacteriocin was partially purified by the adsorption-desorption technique, and the analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed a molecular mass of 3.5 to 6.5 kDa. These data encourage studies on application of this bacteriocin in food systems as an additional hurdle to microbial growth.

"Petit suisse" cheese from kefir: an alternative dessert with microorganisms of probiotic activity

"Petit Suisse" is a creamy cheese. Kefir is a symbiotic mixture of lactic acid bacteria and yeasts with probiotic activity including immunomodulation and balance of intestinal microflora. The present study aims to develop "Petit Suisse" cheese from kefir. Kefir grains were grown in pasteurized cow milk, and after the separation of kefir the serum was discarded and the "Petit Suisse" cheese was prepared using strawberry, mangaba, herbs, and dried tomatoes. The acceptance of the different preparations was evaluated using a nine-point hedonic scale followed by ANOVA. The sweet and salty products were compared by the Student's t-test. Purchase intent was evaluated by the means test and frequency distribution. All products were well accepted by the judges. The product was characterized by low yield, but it can be prepared at home at low cost. The nutritional composition analyses and the variety of flavors as well as the range of age of the judges are alternatives for further studies.

Microbiota of sausages obtained by spontaneous fermentation produced in the South of Brazil

The study of the ecology of fermented sausage is fundamental to understand the physical and chemical changes that happen during fermentation and maturation. The aim of the present study was to determine the microbiological characteristics of sausages produced by spontaneous fermentation. Fifty samples of sausages produced in the South of Brazil by different small manufacturers were analyzed for the following microbiota: aerobic mesophilic bacteria; Micrococcaceae; mold and yeast; lactic acid bacteria; total and fecal coliforms; coagulase-positive Staphylococcus, and Salmonella. In most samples (72%), the count of lactic bacteria was higher than 6 log10 cfu.g-1, and the samples with the highest counts were above 8 log10 cfu.g-1. The counts of Micrococcaceae in most samples were between 5 log10 and 7 log10 cfu.g-1. With respect to the presence of molds and yeasts, there was a significant variation among the samples with counts ranging from 2 log10 cfu.g-1 and 6 log10 cfu.g-1. From the data obtained, it was possible to conclude that 24% of the analyzed samples did not comply with the current law in Brazil since the levels of fecal coliforms or coagulase-positive Staphylococcus exceeded the maximum limit allowed.

Effect of starter culture and inulin addition on microbial viability, texture, and chemical characteristics of whole or skim milk Kefir

The effect of inulin addition and starters (Kefir grains or commercial starter culture) on the microbial viability, texture, and chemical characteristics of Kefir beverages prepared with whole or skim milk was evaluated during refrigerated storage. The type of starter did not influence microbial viability during the storage of the beverages, but the chemical and textural changes (decreases in pH, lactose concentration, and inulin and increased acidity, firmness, and syneresis) were more pronounced in the formulations fermented with grains than those fermented with the starter culture. The addition of inulin did not influence acidity or viability of lactic acid bacteria, but in general, its effect on the survival of acetic acid bacteria, Lactococcus and yeasts, firmness, and syneresis depended on the type of milk and starter culture used. Generally, the yeast, acetic acid bacteria, and Leuconostoc counts increased or remained unchanged, while the total population of lactic acid bacteria and Lactococcus were either reduced by 1 to 2 logs or remained unchanged during storage.

Physicochemical and microbiological evaluation of corrientes artisanal cheese during ripening

The aim of this study was to evaluate some physical and chemical parameters (total solids, pH, acidity, fat, acid degree value of fat, salt, protein and nitrogen fractions) and their effects on the beneficial (lactic acid bacteria: LAB) and undesirable microbial populations (coliforms, Escherichia coli, Staphylococcus aureus, moulds, and yeast) during ripening of Artisanal Corrientes Cheese, an Argentinian cow's milk variety, to determine whether a longer ripening period than usual improve its hygienic-sanitary quality. The protein content was much higher than that of other cow's milk cheeses with similar values of fat. The larger peptides showed values three times higher in the 30 day-old cheese than those obtained in the beginning of the process. Staphylococcus aureus and Escherichia coli were detected (3.04 ± 1.48 log10 cfu/g of cheese, 2.21 ± 0.84 log10 MPN/g of cheese) even at 15 and 30 days of ripening, respectively. The distribution of three hundred LAB strains classified to the genus level (lactococci:lactobacilli:leuconostocs) was maintained during the ripening period. The high number of LAB in rennet may have contributed to the fermentation as a natural whey starter, unknown source of LAB for this specific cheese so far. The physicochemical changes that occur during ripening were not big enough to inhibit the growth of undesirable microorganisms.

Antioxidant activities of kombucha prepared from three different substrates and changes in content of probiotics during storage

Kombucha is a health-promoting fermented beverage worldwide. The present study compared the free-radical scavenging abilities and total reducing power (TRP) of kombucha prepared from low-cost green tea (LGTK), black tea (BTK), and tea powder (TPK). LGTK had the highest scavenging abilities against 2,2-diphenyl-1-picrylhydrazyl (DPPH), superoxide anion and hydroxyl radicals, while BTK showed the highest TRP. Changes in content of probiotics in LGTK were investigated during storage as well. The number of acetic acid bacteria decreased moderately up to 10 days of storage. The number of lactic acid bacteria (LAB) decreased significantly, and their survival rate was only 0.98% at the 8th day of storage.

Starter cultures used in probiotic dairy product preparation and popular probiotic dairy drinks

Dairy products that contain probiotic bacteria are those that are produced with various fermentation methods, especially lactic acid fermentation, by using starter cultures and those that have various textures and aromas. Fermented dairy products are popular due to their differences in taste and their favourable physiological effects. Today, fermented dairy beverages in general are produced locally by using traditional methods. Recently, due to the increased demand for natural nutrients and probiotic products, fermented dairy beverages have reached a different position and are considered to have an important impact on human health and nutrition. In this article, probiotic bacteria and functional dairy products that are produced by using probiotic bacteria are discussed.

Ability of a Lactobacillus rhamnosus strain cultured in milk whey based medium to bind aflatoxin B1

This study aimed to compare Lactobacillus rhamnosus growth in MRS (de Man, Rogosa and Sharpe) broth and a culture medium containing milk whey (MMW) and to evaluate aflatoxin B1 (AFB1) adsorption capacity by bacterial cells produced in both culture media. L. rhamnosus cells were cultivated in MRS broth and MMW (37 °C, 24 hours), and bacterial cell concentration was determined spectrophotometrically at 600 nm. AFB1 (1 µg/ml) adsorption assays were conducted using 1 x 10(10) non-viable L. rhamnosus cells (121 °C, 15 minutes) at pHs 3.0 and 6.0 and contact time of 60 minutes. AFB1 quantification was performed by High Performance Liquid Chromatography. Bacterial cell concentration in MMW was higher (9.84 log CFU/ml) than that in MRS broth (9.63 log CFU/ml). There were no significant differences between AFB1 binding results at the same pH value (3.0 or 6.0) for the cells cultivated in MRS broth (46.0% and 35.8%, respectively) and in MMW (43.7% and 25.8%, respectively), showing that MMW can adequately replace the MRS broth. Therefore, it can be concluded that the use of L. rhamnosus cells cultivated in MMW offers advantages such as reduction in large scale production costs, improvement of environmental sustainability, and being a practicable alternative for decontamination of food products susceptible to aflatoxin contamination.

Characterization of goat milk and potentially symbiotic non-fat yogurt

Combining prebiotics and probiotic microorganisms improve quality in the formulation of foods. In this paper, the characteristics of goat milk and symbiotic yogurt were studied. Raw goat milk was analyzed and the skimming process was optimized. For the formulation of a potentially non-fat symbiotic yogurt made with skimmed goat milk, inulin, gelatin, sugar, and Streptococcus salivarius subsp. thermophilus, Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus casei subsp. rhamnoshus. Chemical characteristics, acceptability, and viability of lactic acid bacteria and probiotic culture were assessed. The protein and fat content of the raw milk was 2.90 and 3.56 g/100 mL, respectively. The optimum skimming process was obtained at 9,800 rpm and 4 °C for 15 minutes. The product formulated had a protein and fat content of 4.04 to 0.04 g/100 mL, good sensory properties, and acceptability of 95%. The lactic bacteria count was 9 × 10(7) CFU mL- 1, and probiotic culture count was higher than 1 × 10(6) CFU mL- 1, which guarantees their effect and capacity to survive in the digestive tract and spread in the intestine. The yogurt was stable during the 21 days of storage. Therefore, this study shows that goat milk yogurt is an adequate delivery vehicle of the probiotic culture L. casei and inulin.

Peptide separation of commercial fermented milk during refrigerated storage

Milk is an important source of bioactive compounds. Many of these compounds are released during fermentation and refrigerated storage. The aim of this study was to determine the release of peptides by lactic acid bacteria in commercial fermented milk during refrigerated storage. The size and profile of peptides were analyzed by polyacrylamide gel electrophoresis and sizeexclusion HPLC. During electrophoresis, it was observed that the peptides were released from caseins, whereas β-lactoglobulin was the whey protein with the highest degradation. HPLC analysis confirmed the pattern of peptide formation observed in electrophoresis. Two fractions lower than 2 kDa with aromatic amino acids in their structure were separated. These results were consistent with those reported for structures of peptides with antihypertensive activity. Therefore, the presence of aromatic amino acids in the peptide fractions obtained increases the likelihood of finding peptides with such activity in refrigerated commercial fermented milk. In conclusion, during cold storage, peptides with different molecular weights are released and accumulated. This could be due to the action of proteinases and peptidases of the proteolytic system in lactic acid bacteria.

Green and brown propolis: efficient natural biocides for the control of bacterial contamination of alcoholic fermentation of distilled beverage

This study aimed to evaluate the efficiency of natural biocides, brown and green propolis, for the control of bacterial contamination in the production of sugarcane spirit. The treatments consisted of brown and green propolis extracts, ampicillin, and a control and were assessed at the beginning and end of harvest season in ten fermentation cycles. In the microbiological analyses, the lactic acid bacteria were quantified in the inoculum before and after the treatment with biocides, and the viability of yeast cells during fermentation was evaluated. The levels of acids, glycerol, total residual reducing sugars, and ethanol were analyzed for the wine resulting from each fermentation cycle. A reduction in the number of bacterial contaminants in the inoculum in the treatments with the natural biocides was observed, but it did not affect the viability of yeast cells. The control of the contaminants led to the production of higher levels of ethanol and reduced acidity in the wine produced. The results of the use of brown and green propolis to control the growth microorganisms in the fermentation of sugarcane spirit can be of great importance for using alternative strategies to synthetic antibacterials in fermentation processes including other distilled beverage or spirits.

Use of starter cultures isolated from native microbiota of artisanal sausage in the production of Italian sausage

The most promising microorganisms for use as starter cultures are those isolated from the native microbiota of traditional fermented products. The aim of this study was to evaluate the use of lactic acid bacteria selected from the native microbiota of sausages produced by spontaneous fermentation as starter cultures for the production of sausage. Strains of Lactobacillus plantarum 503 and 341 have the potential for use as starter cultures in the manufacture of Italian sausage type. The population of lactic acid bacteria in sausages was >8 log CFU.g-1 during fermentation, which caused the pH to decrease to <4.5. This decrease in pH and the water activity of < 0.90 of sausages ensures the safety and preservation of this product. Sausages produced with these lactic cultures fulfill the requirements for microbiological quality and composition of Italian sausage type. Our results suggest the possibility of using these starter cultures for the production of sausages with peculiar characteristics that contribute to the identity of the product.

Evaluation of Petrifilm™ system compared with traditional methodology in count of indicators of sanitary-hygienic quality and pathogenic microorganisms in sheep milk

The objective of this study was to evaluate the applicability of the Petrifilm™ plates to enumerate microbial groups in sheep milk. Samples of sheep milk (n = 30) were plated simultaneously, to enumerate mesophilic aerobes, total coliforms, lactic acid bacteria, Staphylococcus aureus and Escherichia coli, using convencional reference protocols and Petrifilm™ plates. The results were compared using McNemar's test, linear regression and ANOVA (p < 0,05). The results demonstrated good significant between conventional methodologies and Petrifilm™ plates. Further, the Petrifim™ STX for counting S. aureus had higher recoverability of bacteria compared with the conventional methodology. Based on the results obtained and in view of the ease and rapidity procedures results, Petrifim ™ plates may be considered as alternatives for microbiological testing in sheep milk.

Enrichment of nutritional value of Phyllanthus emblica fruit juice using the probiotic bacterium, Lactobacillus paracasei HII01 mediated fermentation

The fermented herbal juices are capable of curing and preventing diseases and reducing the aging progress. The present study was performed to investigate the fermentation of Phyllanthus emblica fruit by Lactobacillus paracasei HII01 with respect to carbon sources, polyphenols, and antioxidant properties. The physical changes, for instance, color, odor, taste, turbidity and gas formation, throughout the fermentation process was manually monitored. The fermented product was rich in polyphenolic content. The acid content and pH of the product were under the norms of Thai community product standards. Antioxidant properties of the fermented product were proved using ABTS, and FRAP assays. Chelation based study suggested that fermented P. emblica fruit juices are healthy enough to stabilize the oxidized form of the metal ion. The optimum fermentation period was 15 days. All the results supported that studied carbon sources did not interfere with the quality of the product. This report is the prelude study on the use of probiotic starter culture for the production of P. emblica fruit based lactic acid bacteria fermented beverages (LAFB) enriched with bioactive compounds. Further research on the impact of different carbon sources and upstream processes on the quality of LAFB is currently in progress.