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[Methods for the study of microbial communities in fermented foods].

The importance of ecological concepts to understand the presence and growth of microorganisms in foods is presently recognized. The production of fermented foods under controlled conditions and its safety assurance depend on the knowledge and control of their microbiota. Traditional fermented foods are obtained by natural fermentations (in which no inoculum is added) and contain complex microbiotas, which are difficult to describe through the use of traditional microbiological methods. The microbial structure of these foods can be studied with different approaches. One of them consists on the typification of isolated microorganisms with methods based on DNA as RFLP, ribotyping, AFLP, ARDRA and RAPD. In order to detect non-culturable or not yet cultured microorganisms, nucleic acids are directly extracted from foods and the microbial diversity is determined from them. Examples of these techniques are the construction of 16S rDNA clone libraries and fingerprinting techniques, such as DGGE and TGGE. Recent advances on the application of these techniques on the study of fermented foods are presented.

Bacteria↗

Food fermentation: a safety and nutritional assessment. Joint FAO/WHO Workshop on Assessment of Fermentation as a Household Technology for Improving Food Safety.

An assessment of the food-safety and nutritional aspects of lactic acid fermentation for the preparation of weaning food at the household level was carried out during a Joint FAO/WHO Workshop held in Pretoria, South Africa, in December 1995. In particular, lactic acid fermentation was evaluated as a part of food preparation processes involving other operations such as soaking, cooking, and the germination of cereal grains. The use of germinated cereals is of particular interest since they can be used to prepare semi-liquid porridges of high nutrient density. After reviewing the present state of knowledge concerning the antimicrobial effects of the lactic acid in fermented foods, and the nutritional benefits of fermentation and the use of germinated cereals, the Workshop made an inventory of gaps in current knowledge and priorities for further research. High priority areas for research include the following: the effect of lactic acid fermentation on viruses, parasites, certain bacteria, and mycotoxins; certain physiological and nutritional effects of the consumption of fermented foods; the characterization and optimization of fermentation processes and the development of appropriate fermentation starters; and risk mitigation using the Hazard Analysis Critical Control Point system, the health education of food handlers, and efforts to change the consumer perception of fermented foods.

Bacteria↗

Survival of bacterial enteric pathogens in traditional fermented foods.

The survival of strains of bacterial enteric pathogens was investigated in two traditional fermented foods (mahewu and sour porridge) and in unfermented porridge. The foods were inoculated with cell suspensions of Salmonella, Shigella, Campylobacter, Aeromonas species and pathogenic Escherichia coli which had a final concentration of 10(6)-10(7) cfu/ml of food. None of the strains of Aeromonas and Campylobacter were detected in mahewu and sour porridge 20 min after inoculation. The salmonellas were not found 4 h after inoculation in either fermented foods but the shigellas and pathogenic E. coli strains were more tolerant to the low pH of the fermented foods. Some of the shigellas and pathogenic E. coli strains survived for 24 h after inoculation but showed a sharp decrease in numbers. All the strains of the enteric pathogens survived for 24 h in the unfermented porridge and increased in the numbers except for campylobacters, the numbers of which declined. These results suggest that the traditional fermented foods have bacteriostatic and bactericidal properties and are unlikely to play a major role in the transmission of bacterial enteric pathogens.

Aeromonas↗

Occurrence and function of yeasts in Asian indigenous fermented foods.

In the Asian region, indigenous fermented foods are important in daily life. In many of these foods, yeasts are predominant and functional during the fermentation. The diversity of foods in which yeasts predominate ranges from leavened bread-like products such as nan and idli, to alcoholic beverages such as rice and palm wines, and condiments such as papads and soy sauce. Although several products are obtained by natural fermentation, the use of traditional starter cultures is widespread. This minireview focuses on the diversity and functionality of yeasts in these products, and on opportunities for research and development.

Asia↗

A review of traditional fermented foods and beverages of Zimbabwe.

Several traditional fermented foods and beverages are produced at household level in Zimbabwe. These include fermented maize porridges (mutwiwa and ilambazi lokubilisa) fermented milk products (mukaka wakakoralamasi and hodzeko) non-alcoholic cereal-based beverages (mahewu, tobwa and mangisi) alcoholic beverages from sorghum or millet malt (doroluthwala and chikokivana) distilled spirits (kachasu) and fermented fruit mashes (makumbi). There are many regional variations to the preparation of each fermented product. Research into the processing technologies of these foods is still in its infancy. It is, therefore, important that the microbiology and biochemistry of these products, as well as their technologies be studied and documented in order to preserve them for future generations. This article reviews the available information regarding traditional fermented foods in Zimbabwe and makes recommendations for potential research areas.

Animals↗

Usage patterns and contribution of fermented foods to the nutrient intakes of low income households in Emene, Nigeria.

This study investigated the usage consumption pattern and chemical composition of fermented foods consumed in 191 rural households (1030 individuals) in Emene. The result showed that fermented foods were widely used and consumed by most age groups (under 2 years to adults) because of poor socioeconomic status. Fermentation period varied with type of food and was mostly carried out as a means of detoxifying certain foods. Generally, fermented foods contributed substantially to the daily caloric (46.3 to 79.9% for males and 57.5 to 78% for females); calcium (33.8 to 63.5% for males and 48.3 to 55.4% for females); iron (34.4 to 58.6% for males and 47.4 to 74.6% for females); and thiamin (23 to 58.5% for males and 37.5 to 60% for females) intakes. The contributions of fermented foods to protein (10 to 40.7%) and ascorbic acid (1.9 to 18.7%) intakes were however, low. When compared with the FAO recommendations, the daily intakes of protein, calcium, riboflavin, niacin and ascorbic acid by the subjects were low due to large consumption of starchy root crops. Poor financial status was the most limiting factor to adequate nutrient intake. Such results point out the need for nutrition education related to improved methods of preparation and food selection.

Adolescent↗

Commercial bacterial starter cultures for fermented foods of the future.

Starter cultures for fermented foods are today developed mainly by design rather than by screening. The design principles are based on knowledge of bacterial metabolism and physiology as well as on the interaction with the food product. In the genomics era, we will obtain a wealth of data making design on a rational basis even simpler. The design tools available are food grade tools for genetic, metabolic and protein engineering and an increased use of laboratory automation and high throughput screening methods. The large body of new data will influence the future patterns of regulation. It is currently difficult to predict in what direction the future regulatory requirements will influence innovation in the food industry. It can either become a promoting force for the practical use of biotechnology to make better and safer products, or it can be limiting the use of starter cultures to a few strains with official approval. Successful cultures based on modern technology is expected to be launched in the areas of: probiotics, bioprotection, general improvement of yield and performance for the existing culture market and probably the introduction of cultures for fermenting other food products. A scientific basis for dramatic innovations that could transform the culture industry is currently being established.

Bacteria↗

Tempeh: a mold-modified indigenous fermented food made from soybeans and/or cereal grains.

A variety of indigenous fermented foods exist today; however, tempeh has been one of the most widely accepted and researched mold-modified fermented products. Tempeh is a traditional fermented food made from soaked and cooked soybeans inoculated with a mold, usually of the genus Rhizopus. After fermentation has occurred, the soybeans are bound together into a compact cake by dense cottony mycelium. An important function of the mold in the fermentation process is the synthesis of enzymes, which hydrolyze soybean constituents and contribute to the development of a desirable texture, flavor, and aroma of the product. Enzymatic hydrolysis also may decrease or eliminate antinutritional constituents; consequently, the nutritional quality of the fermented product may be improved. Current technology and new scientific advancements have enabled researchers to examine specific strains of Rhizopus and new substrates such as cereal grains. Because Kansas produces numerous cereal grains, production of a fermented tempeh-like product using wheat, sorghum (milo), oats, rye, barley, corn, and triticale is a definite possibility for generating a Kansas Value-Added Product. In this study, several different tempeh-like products were produced using various cereal grains inoculated with Rhizopus oligosporus NRRL 2549 or R. oligosporus NRRL 2710. Grains used included hard red winter wheat, triticale, yellow sorghum (milo), and red sorghum (milo). The grain source as well as the strain of R. oligosporus used influenced the product's appearance, flavor, and patty integrity. Results showed that R. oligosporus NRRL 2549 produced more mycelium at a more rapid rate than did the R. oligosporus NRRL 2710 strain. The combination of red sorghum and R. oligosporus NRRL 2549 yielded a product with good patty texture, aroma, and appearance. Furthermore, the red sorghum fermented product was well suited for slicing. On the other hand, yellow sorghum inoculated with either R. oligosporus NRRL 2549 or R. oligosporus NRRL 2710 failed to produce an organoleptically suitable product. Triticale also was found to be an unacceptable substrate for the production of a tempeh-like product. Although the fermented wheat product had a desirable aroma and flavor, it lacked patty integrity and crumbled when sliced. Further research is needed to evaluate the economic significance and industrial applications of these tempeh-like products.

Edible Grain↗

Fermentation, fermented foods and lactose intolerance.

Lactose (milk sugar) is a fermentable substrate. It can be fermented outside of the body to produce cheeses, yoghurts and acidified milks. It can be fermented within the large intestine in those people who have insufficient expression of lactase enzyme on the intestinal mucosa to ferment this disaccharide to its absorbable, simple hexose sugars: glucose and galactose. In this way, the issues of lactose intolerance and of fermented foods are joined. It is only at the extremes of life, in infancy and old age, in which severe and life-threatening consequences from lactose maldigestion may occur. Fermentation as part of food processing can be used for preservation, for liberation of pre-digested nutrients, or to create ethanolic beverages. Almost all cultures and ethnic groups have developed some typical forms of fermented foods. Lessons from fermentation of non-dairy items may be applicable to fermentation of milk, and vice versa.

Dairy Products↗

Studies on bacteriocinogenic Lactobacillus isolates from selected Nigerian fermented foods.

Ten bacteriocin-producing (bacteriocinogenic) Lactobacillus isolates obtained from three Nigerian fermented foods namely: kenkey, ogi and wara were tested against the following indicator organisms: Lactobacillus plantarum and food borne pathogens comprising enterotoxigenic Escherichia coli, enterohaemorrhagic Escherichia coli, Serratia, Pseudomonas, Vibrio cholerae, Aeromonas sobria, Aeromonas cavice, Salmonella typhimurium, Plesiomonas shigelloides and Staphylococcus aureus. All the bacteriocinogenic Lactobacillus were found to inhibit L. plantarum while some inhibited some of the food borne pathogens listed above. The antimicrobial activities of bacteriocins from L. plantarum KKY12 and L. casei OGM12 were caused by proteins detectable in the culture liquids. They are designated Plantacin N and Caseicin A and they have narrow antimicrobial spectra. Plantacin N from L. plantarum KKY12 was active against L. plantarum, Pseudomonas, Aeromonas sobria and Aeromonas cavice whereas Caseicin A from L. casei OGM12 inhibited L. plantarum, enterotoxigenic Escherichia coli and Vibrio cholerae. Caseicin A is stable at 121 degrees C/15 mins, and both are inactivated by proteolytic enzymes. The bacteriocinogenic properties of the local isolates of Lactobacillus can help to reduce hygienic risk and the spoilage of fermented foods.

Bacteriocins↗

Binding activity of natto (a fermented food) and Bacillus natto isolates to mutagenic-carcinogenic heterocyclic amines.

The fermented food, whole meal Natto, viscous polymeric material from Natto, Natto bean, cooked soya bean, and 28 bacterial isolates from Natto were studied for their binding capacity to foodborne mutagenic-carcinogenic heterocyclic amines. The mutagenic heterocyclic amines used were Trp-P-1 (3-amino-1,4-dimethyl-5H-pyrido(4,3-b)indole); Trp-P-2 (3-amino-1-methyl-5H-pyrido(4,3-b)indole); Glu-P-1 (2-amino-6-methyldipyrido(1,2-a:3'2'-d)imidazole); PhIP (2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine); IQ (2-amino-3-methylimidazo(4,5-f)quinoline); MeIQ (2-amino-3,4-dimethylimidazo(4,5-f)quinoxaline); MeIQx (2-amino-3,8-dimethylimidazo(4,5-f)quinoxaline); and MeAalphaC (2-amino-3-methyl-9H-pyrido(2,3)indole). The lyophilized Natto and other fractions of Natto exhibited high binding activity towards Trp-P-1, Trp-P-2, PhIP, and MeAalphaC, while Glu-P-1, IQ, and MeIQ were not effectively bound. The binding capacity of bacterial isolates (Bacillus natto) were isolate-mutagen dependent. Heat treated lyophilized cells, cell wall, and cytoplasmic contents of the bacterial isolate with the highest binding capacity were analyzed for their ability to bind different heterocyclic amines. The results indicate the importance of the cell wall in binding to heterocyclic amines, whereas the cytoplasmic contents were less effective. Heat-treated cells were not much different from that of viable cells in their binding. The impact of different factors, such as pH, incubation time, metal ions, different concentrations of sodium chloride and alcohol, various enzymes, and acetylation of mutagens on binding of Trp-P-1 and IQ, were discussed. The significance of the present results is also discussed from the viewpoint that Natto, a fermented food, is able to scavenge dietary mutagenic heterocyclic amines through binding.

Amines↗

Metabolic engineering of lactic acid bacteria: overview of the approaches and results of pathway rerouting involved in food fermentations.

Lactic acid bacteria such as Lactococcus lactis are the microorganisms of choice for performing metabolic engineering in relation to food fermentation. These bacteria are used extensively in food fermentations, they have a simple and therefore controllable metabolism and the molecular genetics of these food bacteria is well-developed. There have been recent successes in metabolic engineering in these lactic acid bacteria, including examples of changes in both primary metabolism (diacetyl and alanine) and secondary metabolism (exopolysaccharides and flavour).

Alanine↗

Studying the dynamics of microbial populations during food fermentation.

The dynamics of growth, survival and biochemical activity of microorganisms in food are the result of stress reactions in response to the changing of the physical and chemical conditions into the food microenvironment, the ability to colonise the food matrix and to growth into a spatial heterogeneity, and the in situ cell-to-cell ecological interactions which often happen in a solid phase. In food, ecological approaches to study the evolution of microbial flora would be useful to comprehend better the microbiological processes involved in food processing and ripening, to improve microbiological safety by monitoring in situ pathogenic bacteria, and to evaluate the effective compositions of the microbial populations. This paper gives a general overview of biotechnological approaches to study microbial populations in food fermentation.

Bacteria↗

Plasmid profiles of bacteriocin-producing Lactobacillus isolates from African fermented foods.

Fourteen of 200 Lactobacillus isolates from African fermented foods, viz. 'wara', 'kenkey', 'ugba', 'ogi', 'kunuzarki', 'fufu' and 'iru' were found to produce bacteriocins against L. plantarum and only three bacteriocinogenic isolates inhibited some of the food pathogens. Plasmid analysis of the 14 bacteriocin-producing lactobacilli showed that only 5 isolates harbored plasmids ranging in size from 3.1 to 55.5 kb.

Africa↗

Anti-allergy properties of fermented foods: an important immunoregulatory mechanism of lactic acid bacteria?

Clinical reports have suggested that dietary consumption of fermented foods, such as yogurt, can alleviate some of the symptoms of atopy and might also reduce the development of allergies, possibly via a mechanism of immune regulation. Controlled studies have indicated that consumption of fermented milk cultures containing lactic acid bacteria (LAB) can enhance production of Type I and Type II interferons at the systemic level. In animal models, LAB have been shown to promote interferon expression, and to reduce allergen-stimulated production of IL-4 and IL-5 in some cases. Recent results have shown that LAB are potent inducers of pro-interferon monokines (IL-12 and IL-18), and that cytokine secretion is stimulated by the interaction of Gram-positive cell wall components with surface receptors of mononuclear phagocytes, via NF-kappa B and STAT signalling pathways. However, it is clear that the extent and quality of LAB-induced immunoregulation is strain-dependent. This review discusses the clinical and laboratory evidence for anti-allergy properties of fermented foods, and proposes a model for the mechanism by which some well-defined strains of immunoregulatory LAB might down-regulate a Th2 allergic phenotype.

Animals↗