Isolation and Characterization of Probiotic LAB from Kimchi and Spontaneously Fermented Teff (Eragrostis tef (Zucc.) Trotter) Batter: Their Effects on Phenolic Content of Teff during Fermentation.

Microbial fermentation is proven to induce molecular transformations and produce bioactive compounds thereby enhancing sensory and nutritional quality of flour-based fermented foods. In this study, lactic acid bacteria (LAB) were isolated from Korean kimchi and Ethiopian fermented teff (Eragrostis tef (Zucc.) Trotter) flour batter. Isolates were identified using 16S rRNA gene sequencing and characterized for various probiotic properties. Few trains were selected for further teff flour batter fermentation and evaluating their effects on phenolic contents and compositions. Out of 200 bacterial isolates, 44 of them showed considerable acid and bile tolerance and 22 were tested positive for protease activity. A large number of the isolates showed antimicrobial activities against Salmonella gallinarium indicator strains. Majority of these probiotic strains belonged to Lactobacillus plantarum and Lactobacillus brevis species. All the strains used for fermentation of teff were able to significantly increase total phenolic contents (TPC). An increase in TPC of up to 7-fold was observed in some strains.

Traditional fermentation of tef injera: Impact on in vitro iron and zinc dialysability.

Tef [Eragrostis tef (Zucc.) Trotter], an ancient cereal mainly produced in Ethiopia, is increasingly getting higher acceptance in the global market because it is gluten free and has high iron content. The aim of this study was to evaluate the in vitro dialysability of Fe and Zn in a backslop fermented gluten free flat bread known as injera. The traditional fermentation caused up to 49-66% reduction of phytic acid (PA). Molar ratios of PA:Fe and PA:Zn decreased from 14 to 1 and from 63 to 19, respectively, after 120h of fermentation. The total soluble fractions of Fe and Zn ranged between 11 and 38% and between 11 and 29%, respectively, after 120h of fermentation. The dialyzable Fe content of the white varieties ranged between 3 and 9% after 120h fermentation while no effect was observed for the brown varieties. The dialyzable Zn ranged between 2 and 11%, with only a clear effect of fermentation in one white variety. Consumption of tef could be a good source of Fe and Zn, but may not provide the absolute recommended daily Fe and Zn intakes.

Influence of sourdough on in vitro starch digestibility and predicted glycemic indices of gluten-free breads.

Gluten-free flours (buckwheat, quinoa, sorghum and teff) were fermented using obligate heterofermentative strain Weissella cibaria MG1 (Wc) and facultative heterofermentative Lactobacillus plantarum FST1.7 (Lp). Starch hydrolysis of breads with and without sourdough (controls) was analyzed in vitro using enzymatic digestion followed by dialysis (10-11 kDa). Hydrolysis indices as well as predicted glycemic indices (pGI) were calculated from reducing sugars released into the dialysate. Amounts of resistant starch (RS; % of total starch) were determined by enzymatic digestion. Upon sourdough addition, RS significantly decreased in buckwheat (Wc 1.28%, Lp 1.44%) and teff sourdough breads (Wc 0.87%, Lp 0.98%) in comparison to their controls (2.01% and 1.92%, respectively). However, no correlation was found with starch hydrolysis. Predicted GIs were reduced upon sourdough addition in wheat (ctrl 100; Wc 85; Lp 76) in comparison to control breads. This was not the case in most gluten-free breads with the exception of sorghum (ctrl 72; Lp 69) and teff sourdough breads (ctrl 74; Lp 68). In contrast, increased pGIs were found in quinoa (ctrl 95; Wc 106; Lp 103) and buckwheat sourdough breads (ctrl 80; Wc 89; Lp 86).

Influence of dextran-producing Weissella cibaria on baking properties and sensory profile of gluten-free and wheat breads.

Breads based on gluten-free buckwheat, quinoa, sorghum and teff flours were produced with addition of 20% sourdough fermented with exopolysaccharide (EPS) producing Weissella cibaria MG1. Wheat bread was baked as a reference. Dough rheology, bread quality parameters and sensory properties of the sourdough-containing breads were compared to sourdough non-containing control breads of the respective flour. The specific volume remained unaffected by sourdough application. In buckwheat, sorghum, teff and wheat sourdough breads acidification increased crumb porosity compared to control breads. Crumb hardness was significantly reduced in buckwheat (-122%), teff (-29%), quinoa (-21%) and wheat sourdough breads (-122%). The staling rate was significantly reduced in buckwheat, teff and wheat sourdough breads. Water activity of the sourdough containing bread crumb was not influenced by the presence of EPS. Due to the presence of exopolysaccharides (EPS) and influence of acidification, the dough strength, AF, as measured by oscillation tests decreased significantly in sourdough-containing buckwheat, sorghum and wheat dough, but increased in sourdough-containing quinoa and teff dough. Microbial shelf-life was significantly prolonged neither for gluten-free sourdough nor for wheat sourdough breads. Scanning electron microscopy of control and sourdough bread crumbs did not show differences concerning structural starch features. In addition, the aroma of most bread was not improved by sourdough addition.

Biodiversity of lactic acid bacteria and yeasts in spontaneously-fermented buckwheat and teff sourdoughs.

In this study, four different laboratory scale gluten-free (GF) sourdoughs were developed from buckwheat or teff flours. The fermentations were initiated by the spontaneous biota of the flours and developed under two technological conditions (A and B). Sourdoughs were propagated by continuous back-slopping until the stability was reached. The composition of the stable biota occurring in each sourdough was assessed using both culture-dependent and -independent techniques. Overall, a broad spectrum of lactic acid bacteria (LAB) and yeasts species, belonging mainly to the genera Lactobacillus, Pediococcus, Leuconostoc, Kazachstania and Candida, were identified in the stable sourdoughs. Buckwheat and teff sourdoughs were dominated mainly by obligate or facultative heterofermentative LAB, which are commonly associated with traditional wheat or rye sourdoughs. However, the spontaneous fermentation of the GF flours resulted also in the selection of species which are not consider endemic to traditional sourdoughs, i.e. Pediococcus pentosaceus, Leuconostoc holzapfelii, Lactobacillus gallinarum, Lactobacillus vaginalis, Lactobacillus sakei, Lactobacillus graminis and Weissella cibaria. In general, the composition of the stable biota was strongly affected by the fermentation conditions, whilst Lactobacillus plantarum dominated in all buckwheat sourdoughs. Lactobacillus pontis is described for the first time as dominant species in teff sourdough. Among yeasts, Saccharomyces cerevisiae and Candida glabrata dominated teff sourdoughs, whereas the solely Kazachstania barnetti was isolated in buckwheat sourdough developed under condition A. This study allowed the identification and isolation of LAB and yeasts species which are highly competitive during fermentation of buckwheat or teff flours. Representatives of these species can be selected as starters for the production of sourdough destined to GF bread production.

Development of buckwheat and teff sourdoughs with the use of commercial starters.

In this study we investigated the suitability of commercial starters for the production of gluten free sourdoughs. For this purpose, four different laboratory scale sourdoughs were developed from the flours buckwheat or teff. Two different starters (SA, SB) were used to start the fermentations, which were carried out under two technological conditions. Sourdoughs were propagated by back-slopping until the stability was reached. The composition of the stable sourdoughs was investigated by culture dependent techniques and the development of the dominant biota was monitored by PCR-DGGE. Unique and complex LAB and yeasts communities were detected in each sourdough, comprising strains which originated from the flours. The competitiveness of the starter LAB varied according to the substrate and the fermentation conditions applied. Among the LAB present in both SA and SB, L. helveticus and L. paracasei strains did not persist in buckwheat or teff sourdoughs. Lc. argentinum was competitive only in buckwheat sourdoughs, whereas L. reuteri persisted only in teff sourdough. L. fermentum and L. helveticus present in both starters dominated only the sourdoughs fermented at the higher temperature. Remarkably, the starter yeasts were outcompeted by spontaneous yeast strains, i.e. Kazachstania barnetti and Saccharomyces cerevisiae in teff sourdoughs, whereas no yeasts were isolated from buckwheat sourdoughs. The isolation of autochthonous LAB and yeasts from the stable teff and buckwheat sourdoughs indicates that both flours represent an important reservoir for the isolation of novel and competitive starters for the production of gluten free sourdough bread.

Claviceps cyperi, a new cause of severe ergotism in dairy cattle consuming maize silage and teff hay contaminated with ergotised Cyperus esculentus (nut sedge) on the Highveld of South Africa.

During December/January 1996/97 typical summer syndrome (hyperthermia and a 30% drop in milk yield) occurred in succession in two Holstein dairy herds (n=240 and n=150 milking cows, respectively) on the South African Highveld. These farms are situated in the midst of the prime maize and dairy farming areas of South Africa where this condition had never been diagnosed before. The individual components of the concentrate on both farms were negative for ergot alkaloids. Endophytic fungi and/or ergot infestation of teff and other grasses fed to the cows were then suspected of being involved, but neither endophytes nor ergot alkaloids could be implicated from these sources. By measuring the serum prolactin levels of groups of sheep (n=5) fed the first farm's total mixed ration (TMR) or its three individual fibre components for a period of 11 days, the source of the ergot alkaloids was identified. A statistically significant decrease in the level of this hormone occurred only in the group on maize silage (which constituted 28% on dry matter base of the TMR). The involvement of the maize silage was further chemically confirmed by the high levels of total ergot alkaloids, predominantly ergocryptine, found by LC-MS in the silage as well as in the TMR (115-975 ppb and 65-300 ppb, respectively). The ergot alkaloid content (mainly ergocryptine) of the maize silage on the second affected farm was 875 ppb. Withdrawal of contaminated silage resulted in gradual recovery of stock on both farms. Nut sedge (Cyperus esculentus and Cyperus rotundus of the family Cyperaceae) has a world-wide distribution and is a common weed in annual crops, and can be parasitized by Claviceps cyperi. Careful examination of the maize silage from both farms revealed that it was heavily contaminated with nut sedge and that it contained minute sclerotia, identified as those of Claviceps cyperi, originating from the latter. Nut sedge was abundant on both farms and it is believed that late seasonal rain had resulted in mature, heavily ergotised nut sedge being cut with the silage. Claviceps cyperi sclerotia, collected on the affected fields in the following autumn contained 3600-4000 ppm ergocryptine. That the dominant alkaloid produced by this particular fungus was indeed ergocryptine, was confirmed by negative ion chemical ionization MS/MS. In one further outbreak in another Holstein herd, teff hay contaminated with ergotised nut sedge and containing 1200 ppb alkaloids, was incriminated as the cause of the condition. This is the first report of bovine ergotism not associated with the Poaceae infected with Claviceps purpureum or endophytes but with the family Cyperaceae and this particular fungal phytopathogen.