Industrial wastewaters and dewatered sludge: rich nutrient source for production and formulation of biocontrol agent, <Emphasis Type="Italic">Trichoderma viride</Emphasis>

Axenic cultivation of biocontrol fungus Trichoderma viride was conducted on a synthetic medium and different wastewaters and wastewater sludges in shake flasks to search for a suitable raw material resulting in higher biocontrol activity. Soluble starch based synthetic medium, dewatered municipal sludge, cheese industry wastewater sludge, pre-treated and untreated pulp and paper industry wastewater and slaughter house wastewater (SHW) were tested for T. viride conidia and protease enzyme production. The maximum conidia production followed the order, soluble starch medium (>10⁹ c.f.u./mL), untreated pulp and paper industry wastewater (4.9 × 10⁷ c.f.u./mL) > cheese industry wastewater (1.88 × 10⁷ c.f.u./mL) ≈ SHW (1.63 × 10⁷ c.f.u./mL) > dewatered municipal sludge (3.5 × 10⁶ c.f.u./mL) > pre-treated pulp and paper industry wastewater (1.55 × 10⁶ c.f.u./mL). The protease activity of T. viride was particularly higher in slaughterhouse wastewater (2.14 IU/mL) and dewatered municipal sludge (1.94 IU/mL). The entomotoxicity of soluble starch based synthetic medium was lower (≈6090 SBU/μL) in contrast to other raw materials. The entomotoxicity inversely decreased with carbon to nitrogen ratio in the growth medium and the conidia concentration and protease activity also contributed to the entomotoxicity. The residual c.f.u./g formulation of T. viride conidia were up to approximately, 90% after 1 month at 4 ± 1 °C and about 70% after 6 months at 25 ± 1 °C. Thus, production of T. viride conidia would help in marketability of low cost biopesticide from the sludge and safe reduction of pollution load.     

Measurement of particle diameter of <Emphasis Type="Italic">Lactobacillus acidophilus</Emphasis> microcapsule by spray drying and analysis on its microstructure

Lactobacillus acidophilus, as a probiotic, is widely used in many functional food products. Microencapsulation not only increases the survival rate of L. acidophilus during storage and extends the shelf-life of its products, but also optimal size microcapsule makes L. acidophilus have an excellent dispersability in final products. In this paper, L. acidophilus was microencapsulated using spray drying (inlet air temperature of 170°C; outlet air temperature of 85–90°C). The wall materials used in this study were β-cyclodextrin and acacia gum in the proportion of 9:1 (w/w), and microcapsules were prepared at four levels of wall materials (15, 20, 25 and 30% [w/v]) with a core material concentration of 6% (v/v). The microcapsule diameters were measured by Malvern’s Mastersizer-2000 particle size analyzer. The results showed that the particle diameters of microcapsule were mostly within 6.607 μm and 60.256 μm and varied with 2.884–120.226 μm (the standard smaller microcapsule designated as <350 μm). Through comparison of microcapsule size and uniformity with different concentration of wall materials, we concluded that the optimal concentration of wall material was 20% (w/v), which gave microcapsule with a relatively uniform size (averaging 22.153 μm), and the number of surviving encapsulated L. acidophilus was 1.50 × 10⁹ c.f.u./ml. After 8 weeks storage at 4°C, the live bacterial number was above 10⁷ c.f.u./ml, compared with unencapsulated L. acidophilus, 10⁴–10⁵ c.f.u./ml. Through the observation of scanning electron microscopy, we found that the shapes of microcapsule were round and oval, and L. acidophilus cells located in the centre of microcapsule.     

Evaluation of Azotobacter and Azospirillum as Biofertilizers in Aquaculture

Summary A preliminary comparative evaluation of the two commonly encountered free-living nitrogen fixers in the aquatic system, Azotobacter and Azospirillum was carried out in the laboratory for use as biofertilizers in aquaculture considering the importance of eco-friendly and econo-friendly productivity optimization of freshwater aquaculture. The ammonium–nitrogen levels in water media in Azotobacter treatment varied in the range 2.59–34.34 μg-N/l and was found to be significantly different from that of Azospirillum treatment (p < 0.05). The viable population of the respective nitrogen fixers as colony forming units (c.f.u.) in water media in charcoal-immobilized Azotobacter treatment ranged from 0.39 to 2.48 × 10³ c.f.u./ml and were significantly higher (p < 0.05) than the counterparts. The nitrogenase activity in the same treatment similarly remained higher, at 8.3–12.15 nmol of ethylene/ml water/h followed by the alginate-immobilized Azotobacter treatment which was at 7.2–11.40 nmol of ethylene/ml water/h compared to 5.8–7.8 and 4.65–4.83 in the respective Azospirillum-treated counterparts. Hence, better performance of Azotobacter sp. over Azospirillum sp., and of charcoal immobilization over alginate immobilization were observed.     

Microbial population dynamics, especially stress tolerant Bacillus thuringiensis, in partially anaerobic rice field soils during post-harvest period of the Himalayan, island, brackish water and coastal habitats of India

Population ( × 10⁷ c.f.u./g dr. soil) of the aerobic (30.5–154.1) and anaerobic (5.9–91.4) heterotrophic, aerobic (24.0–56.0) and anaerobic (2.4–4.2) spore forming, Gram (-)ve (1.6–2.9), phosphate solubilizing (10–20), asymbiotic N₂-fixing (0.5–0.9), sulfur oxidizing (1.1–2.0), nitrifying (1.0–5.8) and denitrifying (12.1–18.7) bacteria; as well as, the actinomycetes (about 10⁴ c.f.u./g dr. soil) and fungi (about 10⁵ c.f.u./g dr. soil) were variable in the partially anaerobic, saline and drain out flooded rice soils during the post harvest period of the Himalayan, brackish water flooded, island and coastal habitats of India. The aerobic heterotrophic and spore forming bacteria were more than the anaerobic counterparts in the soils. Population (0.51–3.51 × 10⁶ c.f.u./g dr. soil) and crystal morphotype (spherical, bipyramidal and polymorphic) of the Bacillus thuringiensis (Bt) isolates of different soils were variable. Bt index was 0.002 at Mahe but 0.006 in other soils. The Bt isolates tolerated 5–12% NaCl. The osmolytes (mg/g dr. wt.) like the amino acids (0.38–99.45) and proline (0.38–0.80); and the antioxidative enzymes (units (U)/mg protein/min) viz. the catalase (0.17–5.59) and superoxide dismutase (0.35–74.46) were related with intrinsic osmotic stress tolerance of the Bt but they formed spores to overcome anoxic stress. Two Bt isolates were potent tolerant to both osmotic and anoxic stresses.     

Biological control of root rot fungus <Emphasis Type="Italic">Macrophomina phaseolina</Emphasis> and growth enhancement of <Emphasis Type="Italic">Pinus roxburghii</Emphasis> (Sarg.) by rhizosphere competent <Emphasis Type="Italic">Bacillus subtilis</Emphasis> BN1

Bacterial isolates having antifungal and good plant growth-promoting attributes were isolated from chir-pine (Pinus roxburghii) rhizosphere. An isolate, Bacillus subtilis BN1 exhibited strong antagonistic activity against Macrophomina phaseolina, and other phytopathogens including Fusarium oxysporum and Rhizoctonia solani. It was characterized and selected for the present studies. BN1 resulted in vacuolation, hyphal squeezing, swelling, abnormal branching and lysis of mycelia. The cell-free culture filtrate of BN1 inhibited the growth of M. phaseolina. Pot trial study resulted in statistically significant increase in seedling biomass besides reduction in root rot symptoms in chir-pine seedlings. BN1 treatment resulted in 43.6% and 93.54% increases in root and shoot dry weights respectively, as compared to control. Also, 80–85% seed viability was recorded in treatments receiving BN1 either alone or in the presence of M. phaseolina, compared to 54.5% with M. phaseolina. Bioinoculant formulation study suggested that maximum viability of bacteria was in a sawdust-based carrier. B. subtilis BN1 produced lytic enzymes, chitinase and β-1,3-glucanase, which are known to cause hyphal degradation and digestion of the cell wall component of M. phaseolina. In the presence of M. phaseolina, population of B1 was 1.5 × 10⁴ c.f.u. g⁻¹ root after one month, which increased to 4.5 × 10⁴ c.f.u. g⁻¹ root in three months. Positive root colonization capability of B. subtilis BN1 proved it as a potent biocontrol agent.     

Detection of Viable Toxigenic Vibrio cholerae from Environmental Water Sources by Direct Cell Duplex PCR Assay

Summary Environmental monitoring is important to enable effective resource management and public health protection as well as rapid and accurate identification of Vibrio cholerae in drinking-water sources. Traditional methods employed in identification are laborious, time-consuming and practically not viable for screening of a large number of samples. In this study, a direct cell duplex PCR assay for the detection of viable toxigenic V. cholerae in environmental water samples was developed. In the PCR assay, two gene sequences were amplified together, one of outer membrane protein (ompW), which is species-specific and another of cholera toxin (ctxAB). The detection limit of duplex PCR was 5 × 10⁴ V. cholerae/reaction. Different environmental water samples were artificially spiked with V. cholerae O1 cells and filtered through a 0.22 μm membrane, and the filters enriched in alkaline peptone water for 6 h and then used directly in the duplex PCR assay. The PCR procedure coupled with enrichment could detect as few as 1.2 c.f.u./ml in ground water, 1.2 × 10² c.f.u. ml⁻¹ in sewer water and 1.2 × 10³c.f.u. ml⁻¹ in tap water. The assay was successfully applied directly for screening of environmental potable water samples collected from a cholera-affected area. The proposed method is simple and can be used for environmental monitoring of toxigenic as well as non-toxigenic V. cholerae.     

Production,formulation and antagonistic activity of the biocontrol like-yeast <Emphasis Type="Italic">Aureobasidium pullulans</Emphasis> against <Emphasis Type="Italic">Penicillium expansum</Emphasis>

Aureobasidium pullulans (de Bary) Arnaud (Ach 1-1) was grown in a glucose fed-batch fermentor to 106 g dry wt l⁻¹ in 48 h. The cells were dried in a fluidized bed dryer with a final viability of 62%. After 7 months at 4°C, the viability was 28% of the initial value (= 2.3 × 10¹⁰ c.f.u. g⁻¹ dry matter). A protection level of 89% was achieved with the biomass preparation at 1 × 10⁸ c.f.u. ml⁻¹ after 28 and 7 days for apples stored respectively at 5 and 25°C against Penicillium expansum. Our process is suitable to produce large quantities of the strain Ach 1-1 as biological control agent for apple preservation.     

Water-soluble granules containing <Emphasis Type="Italic">Bacillus megaterium</Emphasis> for biological control of rice sheath blight: formulation,bacterial viability and efficacy testing

Endospores of B. megaterium were formulated in granule formulations with sodium alginate, lactose and polyvinylpyrrolidone (PVP K-30) by the wet granulation technique. The granule formulation exhibited good physical characteristics, such as high-water solubility and optimal viscosity, that would be suitable for spray application. The bacteria remained viable in the dry granule formulation at 10⁹ c.f.u./g after 24 months storage at room temperature. Under laboratory conditions, aqueous solutions of the formulation showed high activity against mycelial growth of R. solani (99.64 ± 0.14% mycelial inhibition). High viability of the bacterial antagonist on leaf sheath and leaf blade at day 7 after spraying with the formulation was observed (approximately 10⁶ c.f.u./g of plant). Application of an equivalent number of un-formulated endospores resulted in much loss of the bacterial endospores even 1 day after application. In a small pilot field study, an aqueous solution of the formulation (3%w/v) applied by spraying at days 1, 5 and 10 after pathogen inoculation of the rice plants was more effective in suppressing rice sheath blight disease than one application of a fungicide (Iprodione) at day 1. Additionally, rice plants sprayed with the aqueous solution of the granule formulation had higher panicle and whole kernel weights than those of fungicide-treated and control (untreated) plants.     

Detection of living <Emphasis Type="Italic">Salmonella</Emphasis> cells using bioluminescence

ATP-based bioluminescence using mutant firefly luciferase was combined with an immunochromatographic lateral flow test strip assay for Salmonella enteritidis detection. In this combination method, the Salmonella-antibody–gold complex captured at the test line on the test strip was lysed by heat-treatment, and the ATP released from the cells was measured using mutant luciferase. This method resulted in approximately 1,000 times higher sensitivity in the detection of Salmonella (i.e. 10³ c.f.u./ml) compared to immunochromatographic lateral flow assay.     

Microbial flora and some chemical properties of ersho,a starter for teff (Eragrostis tef) fermentation

Ersho is a clear, yellow liquid that accumulates on the surface of fermenting teff-flour batter and is collected to serve as an inoculum for the next fermentation. The pH of ersho samples was about 3.5 and titratable acidity ranged between 3.1% and 5.7%. The mean aerobic mesophilic counts from four households varied between 6.9×10⁶ and 1.3×10⁸ c.f.u./ml and the aerobic bacterial flora consisted of Bacillus spp. Mean yeast counts ranged between 5.2×10⁵ and 1.8×10⁶ c.f.u./ml and comprised, in order of abundance, Candida milleri, Rhodotorula mucilaginosa, Kluyveromyces marxianus, Pichia naganishii and Debaromyces hansenii. Candida milleri was the most dominant isolate in all samples. About 90% of the teff flour samples had aerobic mesophilic counts 10⁵ c.f.u./g and Gram-positive bacteria constituted about 71% of the total isolates. About 80% of samples had Enterobacteriaceae counts of 10⁴ c.f.u./g.M. Ashenafi is with the Department of Basic Sciences, Awassa College of Agriculture, Addis Ababa University, PO Box 5, Awassa, Sidamo, Ethiopia.     

Bioreactor for solid-state cultivation of <Emphasis Type="Italic">Phlebiopsis gigantea</Emphasis>

Phlebiopsis gigantea fungus used in biological control of root rot is currently cultivated commercially in disposable, sterilizable plastic bags. A novel packed bed bioreactor was designed for cultivating P. gigantea and compared to the plastic bag method and to a tray bioreactor. The spore viability of 5.4 × 10⁶ c.f.u./g obtained with the packed bed bioreactor was of the same order of magnitude as the viabilities obtained with the other cultivation methods. Furthermore, the packed bed bioreactor was less time and space consuming and easier to operate than the tray bioreactor.     

Selection of rhizosphere-competent Pseudomonas strains as biocontrol agents in tropical soils

Pseudomonas strains were isolated from the rhizosphere of maize grown in yellow-red latosol from Rio de Janeiro, Brazil, to serve as a delivery system for heterologous genes and for risk assessment studies in tropical soils. Selected strains were modified by insertion of the cryIVB gene from Bacillus thuringiensis and tested for pathogenicity gene expression against larvae of a susceptible model species, Anopheles aquasalis. Modified strains Br8 and Br12 showed similar survival performance to their parental strains, and presented a viable density of 10⁷ c.f.u./g dry soil 30 days after release. A strain of P. fluorescens (Br12) that presented positive results for gene expression and the best survival performance, was selected for risk assessment studies in soil microcosms.     

Optimization of a fermentation process for bioinsecticide production by <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis>

In an attempt to develop a cost-effective process for bioinsecticide production by B. thuringiensis, the feeding regime during aerobic cultivation of the bacterium was investigated and optimized. The process was designed as a two-stage process; a first stage of active growth, where glucose and other nutrients were adequately supplied to the growing cells over 12 h, followed by a second stage of 2 h for spore formation and toxin release. In order to maximize spore and toxin yield and productivity, different quantities of glucose and nutrients were fed separately to the growing cells in four different fermentation runs. In all runs, glucose was converted to bacterial biomass during the first stage and subsequently to spores and crystal protein during the second phase. The best results were obtained with a fermentation run supplied with 190 g glucose in 1500 ml. Up to 20.1 g of bacterial insecticides/l were recovered from fermentation broth with a glucose to toxin conversion yield of 0.159 g/g. Also, a markedly high spore concentration of 2.31 × 10¹² c.f.u./ml was obtained. The spore–crystal protein mixture obtained was tested for its insecticidal activity against three of the most agronomically important pests. Among the bioinsecticide-treated insect pests, Egyptian cotton leafworm, Spodoptera littoralis was the most susceptible pest with the lowest LC₅₀ of the bioinsecticides against its larval instar and the highest virulence against adults emerged later on from the surviving larvae.     

Fate of Salmonella typhimurium DT 104 during the Fermentation of ‘Siljo’, a Traditional Ethiopian Fermented Legume Condiment, and during Product Storage at Ambient and Refrigeration Temperatures

Summary The survival of Salmonella spp. in fermenting foods and their elimination during prolonged fermentation is documented. This prompted the study to evaluate the fate of Salmonella typhimurium DT 104 during the fermentation of ‘Siljo’, a traditional fermented legume condiment, and during the storage of the fermented product at ambient and cold temperatures. ‘Siljo‘ was made to ferment naturally and the count of lactic acid bacteria reached 9.9 log c.f.u./ml on day 5. The pH dropped from an initial value of 5.8–4.65 during this time. The lactic acid flora was dominated by Leuconostoc spp. At ambient temperature storage (18–22 °C), the product spoiled on day 16. The spoilage was caused by Bacillus spp. At refrigerated storage (4 °C), however, the count of Bacillus spp. was below detectable limits (<1 log c.f.u./ml) until the end of experiment on day 16. When Salmonella typhimurium DT 104 was inoculated into the fermenting gruel at low initial levels (2.8 log c.f.u./ml), the count decreased steadily and the test strain was not detected by enrichment on day 5. At higher initial inoculum level (5.5 log c.f.u./ml), complete elimination was observed on day 7. In a non-fermenting control gruel, count of the test strain increased by about 3 log units on day 7. In another experiment, the fermented product was inoculated with Salmonella typhimurium DT 104 at low or high inoculum levels and stored at ambient and refrigeration temperatures. Under ambient temperature storage, the test strain was not detectable by enrichment on days 3 and 6 at low and high initial inoculum levels, respectively. The count of the test strain did not decrease markedly under refrigerated storage. The results indicated that, at normal contamination levels in the kitchen environment, fermentation for five days would completely eliminate Salmonella typhimurium DT 104 from the fermenting gruel; ambient temperature storage of the product would completely eliminate the pathogen after three days; and its survival is markedly prolonged by cold storage.     

Ubiquitous presence of normally non-culturable endophytic bacteria in field shoot-tips of banana and their gradual activation to quiescent cultivable form in tissue cultures

Exploring the source of quiescent bacteria in tissue-cultured bananas (Musa sp.) we demonstrate here through a combination of bacterial 16S rDNA-based molecular technique, light microscopy and cultivation-based approaches the ubiquitous presence of endophytic bacteria in the field shoots of different genotypes (Grand Naine, Robusta, Dwarf Cavendish, Ney Poovan and exotic accessions) and their widespread prevalence in apparently clean tissue cultures. A portion of field shoot-tips (10–60%) showed cultivable endophytes, especially during rainy season, yielding 10²–10⁵ colony forming units g⁻¹ fresh tissue in ‘Grand Naine’, which overtly expressed on tissue culture medium as well. The rest showed no colony development on diverse bacteriological media but proved PCR^+ve to bacterial primers indicating the presence of normally non-culturable organisms, which was endorsed by microscopic observations. Such endophytes gradually turned cultivable rendering all visibly clean cultures as quiescent bacteria-harboring after a few (2–4) to several (8–20) passages, resulting in as much as 1.7 × 10⁵ – 4.0 × 10⁷ colony forming units g⁻¹ tissue of ‘Grand Naine’ after ten passages, yielding different organisms. This study has thus exposed the ubiquitous and intense association existing between endophytes and bananas, including their quiescent survival in suspension cultures. The effect due to quiescent bacteria in micropropagated stocks could not be generalized. The observations question the fundamental principle of asepsis in plant tissue cultures and bring in new information on plant-endophtye association in vitro with implications in micropropagation, germplasm conservation, cell culture studies and molecular profiling. The possible involvement of unsuspected endophytic bacteria in tissue-culture associated phenomena like habituation and epigenetic and somaclonal variations are discussed.     

Bacterial Antagonist as Seed Treatment to Control Leaf Blight Disease of Bambara Groundnut (Vigna subterranea)

Summay Soil samples were taken from 48 fields in the southern part of Thailand in which either bambara groundnut (Vigna subterranea) or groundnut (Arachis hypogeae) had been planted. Bacillus spp. were isolated using soil dilution plates and heat treatment to screen for endospore-producing bacteria. Among 342 Bacillus spp. isolates tested, 168 isolates were not antagonistic to Bradyrhizobium sp. strain NC-92 using dual culture technique. Further testing found 16 isolates of Bacillus spp. had the ability to inhibit mycelial growth of Rhizoctonia solani, a causal agent of leaf blight of bambara groundnut. Among these isolates, Bacillus spp. isolate TRV 9-5-2 had the greatest activity in anti-microbial tests against R. solani. This isolate was later identified as B. firmus. A powder formulation of B. firmus was developed by mixing bacterial endospores, talcum, sodium carboxymethylcellulose (SCMC) and polyvinylpyrolidone (PVP). The formulations contained bacterial levels ranging from 10⁸ to 10¹⁰ c.f.u./g and the viability of bacteria in all formulations remained high after 1 year storage at room temperature (26–32 °C). All formulations showed satisfactory effectiveness in vitro in suppressing mycelial growth of R. solani using dual culture technique. The application of formulations as seed treatment showed that these formulations did not cause abnormality of seedling shape and had no effect on the germination of bambara groundnut seeds.     

Isolation,characterization and colonization of 1-aminocyclopropane-1-carboxylate deaminase-producing bacteria XG32 and DP24

Two 1-aminocyclopropane-1-carboxylate deaminase-producing bacterial strains (DP24 and XG32) were isolated from surface-sterilized tomato roots and rizhospere soil. The strains were identified as Pseudomonas fluorescens biovar. IV (XG2) and Erwinia herbicola (DP24) by physiological and biochemical tests, and 16S rRNA gene analysis. Both strains showed positive plant growth-promoting activity when inoculated into cucumber (Cucumis sativus), tomato (Lycopersicon esculentum), pepper (Capsicum annuum) and rapeseed (Brassica napus L.). Colonization ability and behavior of these two strains were determined by treating mutant strains with rifampicin and fluorescence in situ hybridization (FISH) assay with rRNA targeted probes, respectively. Both strains were endophytic colonizers of pepper plants. The behavior of the two strains was not identical. Strain XG32 only colonized the root and reached the max level of 27.7 × 10⁷ c.f.u./g (fresh weight), after 12 days postinoculation, while strain DP24 was able to colonize the roots, stems and leaves. The max level was reached at 40.87 × 10⁷ c.f.u./g (fresh weight) in the roots, 17 × 10⁷ c.f.u./g in the stems after 7 days postinoculation and 44.84 × 10⁷ c.f.u./g in the leaves after 12 days postinoculation.     

Microbiological changes in mawè during natural fermentation

Lactic acid bacteria increased from 3.2 × 10⁶ and 1.6 × 10⁷ c.f.u./g (wet wt) to 2 × 10⁹ and 1.6 × 10⁹ c.f.u./g after 12 to 24 h of fermentation of home-produced mawè (a dough produced from dehulled maize) and commercial mawè, respectively. In commercial mawè, the yeast count increased from 1.3 × 10⁵ to 2.5 × 10⁷ c.f.u./g after 48 h of fermentation before decreasing, whereas in the home-produced mawè it increased from 2.5 × 10⁴ to 3.2 × 10⁷ c.f.u./g after 72 h of fermentation; the dominant yeasts were mainly Candida krusei, although C. kefyr, C. glabrata and Saccharomyces cerevisiae were also present. Enterobacteriaceae counts increased slightly during the initial stage ofthe fermentation, but decreased below the detection level after 24 to 48 h. Enterobacter cloacae was mostly found in commercial mawè and Escherichia coli mostly in homeproduced mawè.D.J. Hounhouigan and C.M. Nago are with the Université Nationale du Bénin, Faculté des Sciences Agronomiques, Département de Nutrition et de Sciences Alimentaires, BP 526, Cotonou, Benin; M.J.R. Nout and F.M. Rombouts are with the Agricultural University, Department of Food Science, Bomenweg 2, 6703 HD Wageningen, The Netherlands. J.H. Houben is with Utrecht University, Department of the Science of Foods of Animal Origin, Yalelaan 2, 3508 TD Utrecht, The Netherlands.     

Flavour profile of idli batter prepared from defined microbial starter cultures

Idli is a traditional cereal/legume-based naturally fermented steamed product with a soft and spongy texture which is highly popular and widely consumed as a snack food item in India. The predominant fermentation microflora comprises lactic acid bacteria and yeasts and causes an improvement in the nutritional, textural and flavour characteristics of the final product. The flavour profile of idli batter prepared with initial levels of 2 × 10⁴ c.f.u. g⁻¹ of Candida versatilis CFR 505 and 2 × 10¹ c.f.u. g⁻¹ of Pediococcus pentosaceus CFR 2123 in 500 g idli batter, packed in polyester polylaminate pouches and stored at 30 ± 2 °C was periodically analysed by GC-MS. The desirable flavour compounds such as ketones, diols and acids were found to be present upto 8 days of storage, whereas undesirable flavours like sulphurous and oxazolidone compounds, ethanone and thiazole appeared in the batter subsequent to 6 days of storage. The sensory attributes of idlis (final product) prepared from the stored batter related well to the determined flavour profile. The present study appeared to indicate that the flavour profile of traditional fermented foods can be a reliable qualitative and quantitative parameter for objective assessment. This revised version was published online in July 2006 with corrections to the Cover Date.     

Antibacterial effect of honey on the in vitro and in vivo growth of Escherichia coli

The study investigated the antibacterial effect of honey against pathogenic Escherichia coli. Honey showed inhibitory activity against the growth of E. coli (ATCC 25922) in agar plate assay. In liquid culture (48 h, 37 °C) the growth rate of bacterial cells decreased in the presence of honey (9.6 × 10⁵ c.f.u./ml) compared with sucrose (2.87 × 10⁸ c.f.u./ml). Rats fed with honey and orally inoculated with E. coli excreted significantly (P < 0.05) less bacterial cells in faeces compared to controls. Animals acclimatized to feeding of honey prior to E. coli inoculation showed a significant decrease in excreted bacterial load compared with the group provided with honey after bacterial inoculation. Consumption of honey also enhanced the concentration of short chain fatty acids in the intestine of rats (83 mM) compared with the control group (44.5 mM). The results show that honey possessed significant antibacterial activity against E. coli under in vitro and in vivo conditions, and indicate the potential benefit of consumption of honey regularly on the microbiological constitution of animals feeding on it.