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Production of an exopolysaccharide-containing whey protein concentrate by fermentation of whey.

Using whey as a fermentation medium presents the opportunity to create value-added products. Conditions were developed to partially hydrolyze whey proteins and then ferment partially hydrolyzed whey with Lactobacillus delbrueckii ssp. bulgaricus RR (RR; an EPS-producing bacterium). In preliminary experiments, pasteurized Cheddar cheese whey was treated with Flavourzyme to partially hydrolyze the protein (2 to 13% hydrolyzed). Fermentation (2 L, 38 degrees C, pH 5.0) with RR resulted in EPS levels ranging from 95 to 110 mg of EPS per liter of hydrolyzed whey. There were no significant differences in the amount of EPS produced during fermentations of whey hydrolyzed to varying degrees. Since a high level of hydrolysis was not necessary for increased EPS production, a low level of hydrolysis (2 to 4%) was selected for future work. In scale up experiments, whey was separated and pasteurized, then treated with Flavourzyme to hydrolyze 2 to 4% of the protein. Following protease inactivation, 60 L of partially hydrolyzed whey was fermented at 38 degrees C and pH 5.0. After fermentation, the broth was pasteurized, and bacterial cells were removed using a Sharples continuous centrifuge. The whey was then ultrafiltered and diafiltered to remove lactose and salts, freeze-dried, and milled to a powder. Unfermented hydrolyzed and unhydrolyzed whey controls were processed in the same manner. The EPS-WPC ingredients contained approximately 72% protein and 6% EPS, but they exhibited low protein solubility (65%, pH 7.0; 58%, pH 3.0).

Electrophoresis↗

Effects of BioChlor and Fermenten on microbial protein synthesis in continuous culture fermenters.

Meta analysis models were constructed from a data-set of 15 continuous culture fermenter trials and 118 observations on studies with either BioChlor (n = 23 observations) or Fermenten (n = 95) included at 10 and 3%, respectively, of dietary dry matter (DM) to evaluate effects of the ingredients BioChlor and Fermenten (B/F) on rumen function. Digestibility of crude protein was significantly increased by 11% with B/F treatment. This was reflected in significant increases in digestibility of DM and organic matter (OM) by 3.6 and 7.9%, respectively. Increased amounts of sugar in the diet in the presence of B/F tended to reduce digestibility of non-structural carbohydrates (NSC); however, the net effect on NSC digestion was small. There was no effect of treatment on most individual volatile fatty acids (VFA) or total VFA production. Propionate production, however, was significantly reduced in treated fermenters. The main effect of B/F as well as of starch and soluble fiber when combined with the treatment was to increase propionate production; however, the interaction between B/F treatment and sugar decreased propionate production markedly, resulting in a net decrease. The acetate-to-propionate ratio increased by 6% with B/F, largely as a result of the decrease in propionate. Production of nonammonia nitrogen was 1% less in B/F-treated fermenters, and interactions between treatment and starch, sugar, or soluble fiber were significant. Treated fermenters produced 15.7% more microbial nitrogen, in association with a significant 37% increase in rumen protein digestion. Interactions between treatment and starch, soluble fiber, or sugar influenced these results. The interaction of B/F and sugar resulted in a decrease in undegradable protein N and an increase in microbial nitrogen production. Ammonia nitrogen concentrations were increased by 24.6% in treated fermenters. Efficiency of microbial nitrogen production from DM, OM, or carbohydrate was significantly increased by B/F. Sugar content increased efficiency of microbial protein production/kg of OM digested or carbohydrate digested in the presence of treatment by >10 times the increase that was attributable to the interaction of treatment with starch. Treatment with B/F reduced moles of VFA produced/kg of microbial nitrogen produced by 16%. This effect was also substantially influenced by interactions between B/F and sugar. If the fermenter results are representative of those in vivo, milk production responses to treatment with B/F will depend on amounts of starch, soluble fiber, and, particularly, sugar in diets. Milk production responses will also depend on the quality of protein in the diet and the comparative benefit that increased flux of microbial nitrogen provides. Increased digestibility of OM should allow additional ruminant production benefits.

Amino Acids↗

Ingestion of milk fermented by genetically modified Lactococcus lactis improves the riboflavin status of deficient rats.

Riboflavin deficiency is common in many parts of the world, particularly in developing countries. The use of riboflavin-producing strains in the production of dairy products such as fermented milks, yogurts, and cheeses is feasible and economically attractive because it would decrease the costs involved during conventional vitamin fortification and satisfy consumer demands for healthier foods. The present study was conducted to assess in a rat bioassay the response of administration of milk fermented by modified Lactococcus lactis on the riboflavin status of deficient rats. Rats were fed a riboflavin-deficient diet during 21 d after which this same diet was supplemented with milk fermented by Lactoccus lactis pNZGBAH, a strain that overproduces riboflavin during fermentation. The novel fermented product, with increased levels of riboflavin, was able to eliminate most physiological manifestations of ariboflavinosis, such as stunted growth, elevated erythrocyte glutathione reductase activation coefficient values and hepatomegaly, that were observed using a riboflavin depletion-repletion model, whereas a product fermented with a nonriboflavin-producing strain did not show similar results. A safety assessment of this modified strain was performed by feeding rodents with the modified strain daily for 4 wk. This strain caused no detectable secondary effects. These results pave the way for analyzing the effect of similar riboflavin-overproducing lactic acid bacteria in human trials. The regular consumption of products with increased levels of riboflavin could help prevent deficiencies of this essential vitamin.

Animals↗

Effect of chestnut tannin on fermentation quality, proteolysis, and protein rumen degradability of alfalfa silage.

Two experiments were conducted on alfalfa to investigate the effects of the addition of commercial chestnut hydrolyzable tannin at ensiling on 1) silage fermentation quality in lab-scale silos and protein degradation in the rumen, and 2) silage fermentation quality and proteolysis in bale silages. Wilted alfalfa was prepared with 4 tannin levels (0, 2, 4, and 6% on a dry matter (DM) basis; T0, T1, T2, T3, respectively) and ensiled in lab-scale silos. Silages (33% DM) were analyzed for fermentation quality, protein rumen degradability in situ, and organic matter digestibility in vitro through gas production after 120 d of conservation. Wilted alfalfa containing 0 and 4% tannin (T0 and T2) was harvested at 40% DM (wilting level I) and 53% DM (wilting level II) for bale (600 mm diameter) silage. Silages were analyzed for fermentation quality after 78 d of conservation. All the silages were well fermented with no butyric acid. Lab-scale silages showed reductions in ammonia, nonprotein nitrogen (NPN) and DM losses in T2 and T3 treatments, while the fermentation acid profiles were unaffected. In experiment 1, the untreated silage (T0) had the highest protein degradability after being incubated in the rumen. The addition of tannin reduced crude protein ruminal disappearance in a dose-dependent manner. However, the tannin reduced the organic matter digestibility by 5.1% for all of the tannin addition levels. The tannin positively affected the silage quality in the round bale silages, in particular reducing ammonia and NPN in the lowest wilting level. In both experiments, T2 treatment reduced proteolysis without any influence of DM on the binding reaction and reduced the NPN by 15% in comparison to the control.

Ammonia↗

Absorption of colostral proteins by newborn calves fed unfermented, fermented, or buffered colostrum.

Unfermented, frozen colostrum from the first three postpartum milkings of 10 cows was thawed, pooled, and treated to produce three diets: 1) unfermented, 2) fermented (7 days at 25 to 27 C), and 3) fermented (as in 2) with pH adjusted to match that of unfermented colostrum. Eighteen newborn, unsuckled Holstein calves were assigned randomly to one of the three diets. Colostrum diets were thawed and fed at 0, 8, 16, 24, and 36 h. Blood was sampled at 0, 4, 8, 16, 24, and 48 h. Minimal breakdown of colostral gamma-globulin and immunoglobulin G (IgG) occurred during fermentation. Protein breakdown during fermentation was associated primarily with the casein fraction. Concentrations of gamma-globulin in serum of calves receiving unfermented colostrum were higher than those of calves fed fermented colostrum at all sampling times beyond 0 h. Concentrations of gamma-globulin in serum of calves fed buffered colostrum were intermediate. Concentrations of IgG followed a similar trend. Health problems were not encountered, indicating potential for passive immunization of newborn calves via fermented, buffered colostrum in emergency situations.

Absorption↗

Feeding value of fermented waste milk with or without sodium bicarbonate for dairy calves.

Effects of feeding waste milk from antibiotic-treated cows on growth, feed efficiency, and incidence of scours of dairy calves were studied. Twenty-four newborn Holstein heifer calves were assigned at random to one of the following treatments: 1) fresh normal milk, 2) fresh waste milk, 3) fermented waste milk, or 4) fermented waste milk plus sodium bicarbonate. Means for fat, crude protein, and total solids in normal milk (3.25, 3.05, and 11.84%) were lower than the same components for fresh waste milk (3.82, 3.42, and 12.59%) and fermented waste milk (4.02, 3.42, and 12.74%). Mean pH's for normal milk, fresh waste milk, and fermented waste milk were 6.6, 6.6, and 5.1. Calves were fed colostrum the first 3 days of life, and their respective treatment milk at 10% of body weight for 42 days. Dry calf feed was offered ad libitum beginning on day 4, and water was available at all times. Mean weight gains (kg) and ratios of average dry feed to gain (kg/kg) for the 42-day treatments were: 1) 19.2, .6; 2) 17.6, .6; 3) 19.6, .7; and 4) 20.1, .6. Incidence of scours was measured as number of days that scours were present per calf during the 42 days. Mean scour days for each group were: 1) 2.0, 2) 1.8, 3) 3.0, and 4) 4.8. There were no detrimental effects on calves fed fresh or fermented milk from cows treated with antibiotics. Addition of sodium bicarbonate did not affect acceptance of fermented milk by calves.

Animals↗

In vitro fermentation of sugars, grains, and by-product feeds in relation to initiation of ruminal lactate production.

In vitro fermentations of various sugars, grains, and by-products were conducted to investigate the relationships between soluble carbohydrates and initiation of ruminal lactate production. Fermentation of hexose sugars, both monosaccharides and disaccharides, resulted in greater accumulation of lactate than did fermentation of pentoses. Results of fermentation of grains and by-products, in order of greatest to least potential to produce lactic acid, were steam-flaked barley = barley = wheat greater than moisture corn = sorghum grain. Water-soluble fractions of the grains and by-products were more rapidly fermented to lactate than the insoluble fractions. Combining 10% soluble fractions from wheat, barley, and steam-flaked barley with 90% insoluble fractions from corn resulted in significant increases in lactate concentration. Analysis of sugar composition of these water soluble materials indicated that monosaccharides and disaccharides constitute 23 to 46% by weight. Data suggest that water-soluble materials play a role in initiation of lactate production from grains, and further production is dependent on starch fermentability.

Animal Feed↗

Response to various amounts of Aspergillus oryzae fermentation extract on ruminal metabolism in cattle.

The objective of this study was to determine whether Aspergillus oryzae fermentation extract stimulated or inhibited ruminal fermentation when fed at higher than recommended doses (3 g/d). Four dietary treatments of A. oryzae fermentation extract were fed daily to six cows fitted with ruminal cannulas. For each of four periods, bromegrass hay (6% CP) with and without extract was fed for 28 d. Dacron bags containing bromegrass cell walls were ruminally incubated to determine ruminal fiber degradation. The A. oryzae fermentation extract did not affect degradation of cell walls, cellulose, or hemicellulose. Total ruminal anaerobic or cellulolytic bacteria were not different among treatments; neither were the proportions of cellulolytic species, Butyrivibrio sp., Ruminococcus albus, or Ruminococcus flavefaciens. Ruminal ammonia was not different; however, total VFA were higher, and pH tended to be lower, when 27 g/d of A. oryzae fermentation extract was fed. The proportion of VFA was not different among treatments. The A. oryzae fermentation extract fed at nine times the recommended dosage did not produce any stimulatory effects, except for total VFA, and was not inhibitory or toxic to ruminal metabolism and forage fiber degradation.

Ammonia↗

Antimutagenic effects of milk fermented by Lactobacillus helveticus L89 and a protease-deficient derivative.

The antimutagenic effects of whey, acetone extracts, and protein fractions isolated from milk that had been fermented by Lactobacillus helveticus L89 were investigated using the mutagen 4-nitroquinoline-N'-oxide in the Ames test (Salmonella typhimurium TA 100). Fermented milk significantly inhibited mutagenesis induced by 4-nitroquinoline-N'-oxide. However, milk fermented by a nonproteolytic variant of the same strain showed no inhibitory effects. Results were similar for the whey fractions and acetone extracts of the fermented milks. After fermentation, milk proteins were fractionated by size-exclusion HPLC and were tested for antimutagenicity. The fraction showing the greatest activity was further analyzed by reverse-phase HPLC. Our results indicate that antimutagenic compounds are produced in milk during fermentation by L. helveticus, and the release of peptides is one possible contributing mechanism.

4-Nitroquinoline-1-oxide↗

Effect of diacetyl on controlling Escherichia coli O157:H7 and Salmonella Typhimurium in the presence of starter culture in a laboratory medium and during meat fermentation.

Diacetyl is a flavor compound that possesses antimicrobial activity and is found in several dairy products. The effect of diacetyl on controlling the growth of two foodborne pathogens, Escherichia coli O157:H7 and Salmonella Typhimurium, when grown with Pediococcus acidilactici as a meat starter culture was evaluated in a laboratory medium and during salami fermentation. Diacetyl (50 ppm) added to each mixed culture system strongly inhibited the growth of E. coli O157:H7 and Salmonella Typhimurium in the laboratory medium (brain heart infusion, 2.3% of NaCl, 0.75% of dextrose) (P < 0.05). During meat fermentation, the growth of E. coli O157:H7 and Salmonella Typhimurium was inhibited significantly by addition of diacetyl (300 ppm) (P < 0.05) after 24 h fermentation. However, the acid production and growth of P. acidilactici were not affected by the addition of diacetyl (P > 0.05). After 24 h meat fermentation, about a 1.0-log CFU/g difference occurred in numbers of each foodborne pathogen mixed with P. acidilactici (P < 0.05) with and without 300 ppm diacetyl. Diacetyl and the acid produced by the meat starter culture reduced the growth of the two foodborne pathogens during salami fermentation. These results suggest that diacetyl can be used as a food ingredient during meat fermentation to control E. coli O157:H7 and Salmonella Typhimurium without harmful effects on the growth and acid production of P. acidilactici.

Animals↗

Reduction of fumonisin B1 and zearalenone by lactic acid bacteria in fermented maize meal.

Fusarium species are fungi that infect maize products worldwide and elaborate mycotoxins, which have been associated with cancer. This study was carried out to investigate the potential of lactic acid bacteria fermentation in reducing mycotoxin concentration and toxicity in maize meal products. Maize meal was spiked separately with fumonisin B1 and zearalenone and then allowed to ferment for 4 days. The potential cytotoxicity of the mycotoxin-spiked fermented extracts was also investigated using the SNO human esophageal carcinoma cell line (the SNO cell line was explanted from a cancer patient, S.N., a 62-year-old Zulu man, in July 1972). A significant decrease (P < 0.05) in the concentration of the two mycotoxins was observed, with a 56 to 67% and a 68 to 75% reduction in the third and fourth days, respectively. The two mycotoxins were not detectable in commercially fermented maize meal (amahewu) samples. After fermentation, mycotoxin-spiked maize meal samples containing lactic acid bacteria culture were comparatively less toxic to SNO cells than were samples without lactic acid bacteria. However, this difference in toxicity was not significant (P > 0.05). These results indicate that lactic acid bacteria fermentation can significantly reduce the concentration of mycotoxins in maize. However, such a reduction may not significantly alter the possible toxic effects of such toxins. The exact mechanism of toxin reduction warrants further investigation.

Consumer Product Safety↗

Survival of Listeria monocytogenes and Escherichia coli O157:H7 during sauerkraut fermentation.

Sauerkraut was produced from shredded cabbage, as is typical in the United States, and from whole head cabbages, which is a traditional process in parts of Eastern Europe. The sauerkraut was inoculated with five strain mixtures of Escherichia coli O157:H7 and Listeria monocytogenes, and the populations of these bacteria, as well as lactic acid bacteria, pH, and titratable acidity, were monitored over the course of fermentation. Fermentation variables were temperature (18 and 22 degrees C) and salt concentration (1.8, 2.25, and 3%). For most of the analyses, the type of cabbage processing was a significant factor, although within cabbage type, neither salt nor fermentation temperature had significant effects. The final pH of the whole-head sauerkraut was lower than the shredded sauerkraut, but the titratable acidity was significantly higher in the shredded sauerkraut. E. coli O157:H7 and L. monocytogenes persisted in the brines for most of the fermentation, although at the end of the fermentations (15 days for shredded, 28 days for whole head), neither pathogen had detectable populations. E. coli populations decreased more rapidly in the shredded sauerkraut even though the pH was higher because of the higher total acidity in the shredded sauerkraut. Acid-tolerant strains of E. coli and L. monocytogenes were isolated from both shredded and whole-head sauerkraut at different salt concentrations and temperatures after 15 days of fermentation and could be detected at 35 days in the wholehead sauerkraut.

Brassica↗

Behavior of enterotoxigenic strains of Staphylococcus aureus in milk fermented with a yogurt starter culture.

The ability of a yogurt starter culture formed by Streptococcus salivarius subsp. thermophilus and Lactobacillus delbrueckii subsp bulgaricus to inhibit the growth of four enterotoxin type A and B producers Staphylococcus aureus strains (ATCC 6538, S6, FRI-100 and a strain isolated from milk) during fermentation of milk and subsequent storage was investigated. Sterile skim milk was inoculated with about 10(6) CFU/ml of S. aureus and with about 10(6) CFU of starter culture, and incubated at 42 degrees C during 8 h, followed by refrigeration at 4 degrees C. Samples were taken every 2 h during fermentation and every 2 days during storage. Viable count of lactic acid bacteria and S. aureus as well as pH, acidity, thermostable deoxyribonuclease (TNase) and staphylococcal enterotoxin A (SEA) production were evaluated. Behavior of four strains was similar; S. aureus survived the 8 h fermentation with LAB, and its population began to decrease from the first day of storage, being completely inhibited at 9-10 days. TNase and SEA production were positive in all samples taken along the study. It was demonstrated that enterotoxigenic strains of S. aureus were able to survive the fermentation of milk with a yogurt starter culture and they were inhibited after several days during storage of the fermented product, contrary to the general belief which considered it very difficult due to the low pH. Even though S. aureus was inhibited, TNase and SEA were demonstrable along the storage. Therefore, fermented milks may play an important role in the transmission of this organism.

Animals↗

Studies related to the scale-up of high-cell-density E. coli fed-batch fermentations using multiparameter flow cytometry: effect of a changing microenvironment with respect to glucose and dissolved oxygen concentration.

Multiparameter flow cytometric techniques developed in our laboratories have been used for the "at-line" study of fed-batch bacterial fermentations. These fermentations were done at two scales, production (20 m(3)) and bench (5 x 10(-3) m(3)). In addition, at the bench scale, experiments were undertaken where the difficulty of achieving good mixing (broth homogeneity), similar to that found at the production scale, was simulated by using a two-compartment model. Flow cytometric analysis of cells in broth samples, based on a dual-staining protocol, has revealed, for the first time, that a progressive change in cell physiological state generally occurs throughout the course of such fermentations. The technique has demonstrated that a changing microenvironment with respect to substrate concentration (glucose and dissolved oxygen tension [DOT]) has a profound effect on cell physiology and hence on viable biomass yield. The relatively poorly mixed conditions in the large-scale fermentor were found to lead to a low biomass yield, but, surprisingly, were associated with a high cell viability (with respect to cytoplasmic membrane permeability) throughout the fermentation. The small-scale fermentation that most clearly mimicked the large-scale heterogeneity (i.e., a region of high glucose concentration and low DOT analogous to a feed zone) gave similar results. On the other hand, the small-scale well-mixed fermentation gave the highest biomass yield, but again, surprisingly, the lowest cell viability. The scaled-down simulations with high DOT throughout and locally low or high glucose gave biomass and viabilities between. Reasons for these results are examined in terms of environmental stress associated with an ever-increasing glucose limitation in the well-mixed case. On the other hand, at the large scale, and to differing degrees in scale-down simulations, cells periodically encounter regions of relatively higher glucose concentration.

Biotechnology↗

A baker's yeast mutant (fil1) with a specific, partially inactivating mutation in adenylate cyclase maintains a high stress resistance during active fermentation and growth.

The initiation of fermentation in the yeast Saccharomyces cerevisiae is associated with a rapid drop in stress resistance. This is disadvantageous for several biotechnological applications, e.g. the preparation of freeze doughs. We have isolated mutants in a laboratory strain which are deficient in fermentation-induced loss of stress resistance ('fil' mutants) using a heat shock selection protocol. We show that the fil1 mutant contains a mutation in the CYR1 gene which encodes adenylate cyclase. It causes a change at position 1682 of glutamate into lysine and results in a tenfold drop in adenylate cyclase activity. The fil1 mutant displays a reduction in the glucose-induced cAMP increase, trehalase activation and loss of heat resistance. Interestingly, the fil1 mutant shows the same growth and fermentation rate as the wild type strain, as opposed to other mutants with reduced activity of the cAMP pathway. Introduction of the fil1 mutation in the vigorous Y55 strain and cultivation of the mutant under pilot scale conditions resulted in a yeast that displayed a higher freeze and drought resistance during active fermentation compared to the wild type Y55 strain. These results show that high stress resistance and high fermentation activity are compatible biological properties. Isolation of fil-type mutations appears a promising avenue for development of industrial yeast strains with improved stress resistance during active fermentation.

Adenylyl Cyclases↗

Effect of highly fermentable dietary fiber on the development of swine dysentery and on pig performance in a "Pure--Culture Challenge Model".

This study tried to evaluate the effect of highly fermentable fiber on the incidence and severity of swine dysentery (SD) after experimental oral infection with pure cultures of Brachyspira (B.) hyodysenteriae. Forty eight growing pigs were allocated to two groups and treated until slaughter as follows: Group 1 (n = 24): infected with B. hyodysenteriae and fed with a food containing 9.6% highly fermentable neutral detergent fiber. Group 2 (n = 24): infected with B. hyodysenteriae and fed with a food containing 6.1% low fermentable neutral detergent fiber. Pigs of each group were intragastrically inoculated on each of three consecutive days with pure culture of 1.8 x 10(10) B. hyodysenteriae. All pigs were monitored daily until slaughter. Faecal shedding of B. hyodysenteriae by polymerase chain reaction, antibody response by IFA, clinical signs, growth performance and extents of gross and microscopical lesions specific for swine dysentery were determined. Faecal shedding of B. hyodysenteriae and antibodies specific for B. hyodysenteriae were detected at day 30 post infectionem. Significant (p < 0.05) milder clinical signs typical for swine dysentery were detected in group 1, fed with 9.6% high fermentable fiber compared to group two fed with a food containing 6.1% low fermentable neutral detergent fiber. Daily weight gain differed significantly (p < 0.05) between the groups (group one 780 g vs. group two 760 g). Food conversion efficiency showed in group one a significant (p < 0.05) better (3.28) result than in group two (3.38). Feed consumption presented significantly (p < 0.001) better results in group one compared to group two (2.38 kg vs. 2.25 kg). From our experimental findings we conclude that in production units suffering of swine dysentery high levels of highly fermentable fiber in diet may increase health and performance.

Animals↗

[Comparison and analysis of hydrogen production capacity with different acidogenic fermentative microflora].

The biogas(hydrogen) production capacity of ethanol type and propionic acid type fermentative microflora were compared, and the movement of hydrogen production rate according to the succession of microflora from propionic acid type to ethanol type was investigated. It was demonstrated that under the same loading rate, microflora of ethanol type fermentation had relatively high hydrogen production rate and specific hydrogen production rate, maximum hydrogen production rate was 14.99 L/d, maximum specific hydrogen production rate was 3586.45 mmol/(kg.d). While, for the microflora of propionic acid type fermentation, hydrogen production rate and specific hydrogen production rate were extremely low, maximum hydrogen production rate was 3.62 L/d, maximum specific hydrogen production rate was 196.46 mmol/(kg.d). In order to obtain high yield of H2 in the fermentation process, ethanol type fermentation was preferred and propionic and type fermentation had to be avoided.

Bacteria↗

Biological utilization of naturally fermented pearl millet flour (Pennisetum typhoideum).

Natural fermentation of pearl millet flour at 20, 25 and 30 degrees for 72 h brought about an improvement in its apparent and true protein digestibility. Utilisable protein, net protein retention and protein retention efficiency values were also enhanced as a result of fermentation. Rats fed on flour fermented at 20 and 25 degrees C had higher food as well as protein efficiency ratios than the flour fermented at 30 degrees C. Feeding of the fermented products did not bring about any histopathological abnormality in rats. Cutlets prepared from the fermented flour were organoleptically acceptable to a panel of judges.

Animals↗