Five samples of Airag and 20 of Tarag (both in Mongolia) were collected from scattered households. One hundred strains of lactic acid bacteria (LAB) were isolated and identified from these samples according to phenotypic characterization and 16S rRNA gene sequence analysis. Eighty-five isolates belonged to the genus Lactobacillus, 15 being classified as coccoid LAB. All isolates belonged to 5 genera and 11 to different species and subspecies. Lactobacillus (Lb.) helveticus was predominant population in Airag samples, Lb. fermentum and Lb. helveticus were the major LAB microflora in Tarag.

Abstract Aim: The aim was to isolate and characterize Lactococcus strains with new properties compared to those of usual Lactococcus dairy starters derived from cow's milk. Methods and Results: Algerian dromedary's milk was screened for proteolytic isolates able to grow rapidly on agar milk medium. PCR experiments revealed that 74 proteolytic isolates belonged to the genus Lactococcus and harboured the prtP gene encoding the lactococcal cell-surface proteinase. Among these, 85% were able to ferment citrate (Cit(+) phenotype) and were classified as Lactococcus lactis ssp. lactis biovar. diacetylactis. This classification was confirmed after sequencing of the 16S rDNA gene of five Cit(+) isolates. In contrast to dairy lactococci described in the literature, several Cit(+) isolates exhibited a tolerance to 50 degrees C (Ther(+)) and alkaline pH. Two genetic approaches allowed to show the presence of four independent plasmids (so-called pTher, pPrt, pLac, pCit) associated with the four respective phenotypes: Ther(+), cell-surface proteinase activity PrtP (PrtP(+)), lactose catabolism (Lac(+)) and citrate utilization (Cit(+)). Two types of pCit plasmid were amplified by inverse PCR: class 1 was characterized by a 9-kb plasmid harbouring the expected lactococcal citQRP operon and class 2 by a 23-kb plasmid harbouring the Leuconostoc cit cluster (citI-CitMCDEFGRP). Conclusions: This work enlarges knowledge of the biovariety diacetylactis by far mainly limited to the citrate-fermenting ability and suggests that the cit plasmid system of some lactococcal strains could have been acquired from another lactic acid bacteria (Leuconostoc spp.). Significance and Impact of the Study: This study reveals new potential dairy lactococci starters of the biovariety diacetylactis able to grow rapidly in milk at a higher temperature in addition to their casein, lactose and citrate-utilizing abilities.

Abstract Aims: Genotypic and technological characterization of wild lactococci isolated from artisanal Manchego cheese during the ripening process for selection of suitable starter cultures. Methods and Results: A total of 114 isolates of lactococci were typed using randomly amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). Sixteen distinct RAPD-PCR patterns, at a similarity level of 73%, were obtained. On the basis of species-specific PCR reaction, the isolates were assigned to the species Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris with L. lactis subsp. lactis being predominant at both dairies. Twenty-six isolates were technologically characterized to select those with the best properties. Most of them showed good technological properties although some could produce tyramine. Conclusions: The presence of coincident genotypes at both dairies has been demonstrated, which would suggest that they are well adapted to the Manchego cheese environment. Interesting differences were found in the technological characterization and the potential role of autochthonous lactococci strains as starter culture has been displayed. Significance and Impact of the Study: The great economic importance of Manchego cheese encouraged a deeper knowledge of its microbiota, to select strains with the best properties to use as starter cultures in industrial Manchego cheeses, preserving the autochthonous characteristics.

The Beni Mellal province has one of the highest incidences of scorpion envenomation in Morocco. The purpose of this retrospective study was to report and analyze data recorded in scorpion envenomation victims admitted to the intensive care department of the Beni Mellal Provincial Hospital. A total of 63 patients were admitted in 2005. Most stings occurred during summer mainly in June and July. Most (60.3%) occurred at night between 6 p.m. and 6 a.m. Most patients were children younger than 16 years (mean, 10.2 +/- 12.9 years). At the time of admission, general manifestations of envenomation were present in 72.6% of patients and life-threatening symptoms were observed in 27.4%. Therapy consisted of symptomatic treatment and in-hospital mortality was 28.6%. There was not a significant difference in mortality in function of age or severity of manifestations at the time of admission.

Lactic acid bacteria (LAB) strains were assayed for the conservation of fresh sardine "Sardina pilchardus". Lactobacillus delbrueckii subsp. delbrueckii was used for inoculation of sardine fillets in a solution of NaCl (5%, w/w) and glucose (4%, w/w) concentration in water. Microbial counts including Standard Plate Count (SPC), LAB, yeasts, coliforms, Salmonella, staphylococci and Clostridium were followed during two weeks of storage at 30 degrees C. Determinations of chemical parameters including pH, dry matter, fat, ash, total nitrogen (NT), total volatile basic nitrogen (TVBN) and trimethylamine (TMA) were carried out under the same conditions. Chemical determinations showed a net pH decrease from an initial value of 6.05 in raw sardine fillets to 4.3 after 16 days of fermentation. Increases in TMA content and TBVN were observed. Microbiological control showed that LAB counts reached a level up to 3.10(9) cfu/g after 4 days of fermentation. After two weeks, fermented fish was free of coliforms and Salmonella. The inhibition of pathogenic microflora including staphylococci and Clostridium was also observed. The results indicated that controlled LAB fermentation could be used as a successful process for biopreservation of sardines produced in huge quantities in Morocco.

Poisonous stings caused by scorpion constitute a public health problem in Morocco because of their frequency severity and socio-economic consequences that they generate. The province of Khouribga surveyed for this study is situated in a zone of high incidence and high lethality caused by scorpion stings in Morocco. In order to analyze the epidemiological profile and the poisoning risk factors, we are presenting the results of a past study conducted from April to December 2001. It consisted in making an exhaustive follow up from admission to release of all patients stung by scorpion. These patients consulted a sanitary structure (dispensary health centre or hospital) in the province of Khouribga, Morocco. Within a total of 1212 cases of stings, 63% of them occurred at night (between 06 pm and 06 am) particularly during July and August. Youngsters under fifteen represented 36% of these cases. 3.4% of the people who got stung presented typical poisoning and envenomation. The delay expected from sting to transfer to a medical structure was 1.85 +/- 0.11 hours for the patients who recovered against 2.67 +/- 0.11 for those who did not recover. To be noticed as well that 1.3% of the subjects who got stung die. Variance analysis indicates that the type of sanitary structures receiving patients, admission classes, and age of patients influence significantly the recovery rate of people who are stung.

The aim of this study was to determine the prevalence and risk factors for subclinical endometritis (SE) and its effects on fertility in repeat breeder dairy cows. Dairy cows of parity 1 to 5 that were artificially inseminated (AI) 3 or more times (selected cows were artificially inseminated an average of 3.9 times) were examined at 190 +/- 40 days in milk, and clinically normal cows (n = 77) were selected based on the absence of abnormal discharges on external inspection and the absence of abnormal findings on transrectal palpation and ultrasonographic examination. Endometrial samples were collected from the uterus by using the lavage technique in the luteal phase of the estrus cycle. Collected samples were centrifuged and a drop of sediment was streaked on to a clean microscopic slide and stained with Giemsa. The percentage of polymorphonuclear cells (neutrophils) was counted for each specimen. The analysed data showed that the average amount of neutrophils was 3.1%(0-9) in the selected cows. Abnormal calving (dystocia, twin births, and abortion), retained placenta, and postpartum uterine infections were associated with an increase in prevalence of SE. Subsequently, SE was significantly associated with a decrease in conception rate in the next AI. Conception rate in the next AI was 5% for cows (n = 38) with SE (>/= 3% neutrophil), and 47% for cows (n = 34) without SE (< 3% neutrophil)(P = 0.001). The prevalence of cytologically diagnosed SE (>/= 3% neutrophil) was 52.7%(n = 38). In conclusion, abnormal calving, retained placenta, and postpartum uterine infections may be associated with an increase in prevalence of SE and subsequently, SE may decrease reproductive performance and increase the incidence of repeat breeder syndrome.

In this work, a batch chemiluminescence (CL) method has been proposed for the simultaneous determination of two structurally similar alkaloids, noscapine and thebaine. The method is based on the kinetic distinction of the CL reactions of noscapine and thebaine with Ru(bipy)(3)(2+) and Ce(IV) system in a sulfuric acid medium. The least squared support vector machine (LS-SVM) regression was applied for relating the concentrations of both compounds to their CL profiles. The parameters of the model consisting of sigma(2) and gamma were optimized by constructing LS-SVM models with all possible combinations of these two parameters to select the model with the minimum root mean squared error of cross validation (RMSECV) as the best. The parameters of this model were then selected as optimized values. Under the optimized experimental conditions for both compounds, the detection limits obtained using the LS-SVM regression were 0.08 and 0.1 micromo lL(-1) for noscapine and thebaine, respectively. The proposed method was utilized for the simultaneous determination of the compounds in pharmaceutical formulations and plasma samples with satisfactory results.

In this study, changes in viability, biomass production, essential oil yield and essential oil composition of Mentha spicata L.(spearmint) exposed to olive mill wastewater (OMW) were investigated. Spearmint cuttings were sensitive to OMW and, after 6h of incubation in raw or diluted OMW, their viability was null. The short contact of raw OMW with mint cuttings caused an irreversible damage in rhizogenesis and shoots development. Roots were more sensitive to phytotoxicity than shoots. In a field essay, spearmint showed a good capability to recover when OMW was spread at 8lm(-2) at the vegetative phase of growth (45 days after plantation). At this dose, a slight increase of mostly of the mint essential oil constituents was obtained. When the dose applied was 16lm(-2), phytotoxicity was manifested by a high reduction of biomass and essential oil yield. The essential oil composition was also affected and a disappearance of many of mint essential oil constituents was observed with an increase of 59% for carvone, the major compound of spearmint essential oil. As far as we know, this is the first report on the effect of field application of OMW on an aromatic plant essential oil yield and composition.

1. The aim of this study was to determine the effects of added dietary vitamin C on the intestinal mucosa morphology in pulmonary hypertensive broiler chickens. 2. One-day-old chickens were reared at high altitude and treated with 400, 800 or 1200 mg vitamin C/l of drinking water for 48 d. Segments from the duodenum, jejunum and ileum were collected for morphometric analysis. 3. The duodenum and jejunum villus height, width, surface area and lamina propria thickness were greater in the highest vitamin C-treated groups compared to controls. 4. There were significant variations only in villus types of the jejunum and ileum: increased proportions of leaf + tongue-like villi and decreased proportions of convoluted + ridge-like villi at day 36 at the groups with 400 and 800 mg vitamin C/l. 5. It was concluded that vitamin C improves the gut morphology of the pulmonary hypertensive broiler chickens.

A new chemiluminescence (CL) method using flow injection has been described for the rapid and sensitive determination of promazine hydrochloride (PMH). The method is based on the CL reaction of PMH with tris(1,10 phenanthroline)ruthenium(II),[Ru(phen)(3)(2+)] and Ce(IV) in sulfuric acid medium. Effects of chemical variables were investigated employing central composite design and response surface methodology. Under the optimum conditions, the CL intensity was proportional to the concentration of the drug in solution over the ranges 0.020-0.32 and 0.32-32 microg/mL. The limit of detection (signal-to-noise ratio = 3) was 0.012 microg/mL. The method was applied successfully to the determination of PMH in drug formulations and human serum (recovery percentages between 96.7 and 105.0%). The relative standard deviation for 11 replicate determinations of 1.5 microg/mL of PMH was 1.7%. The minimum sampling rate was 100 samples per hour. Copyright (c) 2009 John Wiley & Sons, Ltd.

A flow injection chemiluminescent (FI-CL) method has been developed for the simultaneous determination of codeine and noscapine using N-PLS regression. The method is based on the fact that kinetic characteristics of codeine and noscapine are different in the Ru(phen)(3)(2+)-Ce(IV) CL system. In flow injection mode, codeine gives broad peak with the highest CL intensity at 4.4s, whereas the maximum CL intensity of the noscapine appears at about 2.6s. Moreover, the effect of increasing H(2)SO(4) concentration was different on the CL intensity of the compounds. An experimental design, central composite design (CCD), was used to realize the optimized variables such as Ru(II) and Ce(IV) concentrations for the both compounds. At the optimized condition, a three-way data structure (samples, H(2)SO(4) concentration, time) was constructed and followed by N-PLS regression. The number of factors for the N-PLS regression was selected based on the minimum values for the root mean squared error of cross validation (RMSECV). The proposed method is applied to the simultaneous quantification of codeine and noscapine in the pharmaceutical preparations.

AIM: The contribution of host genetic factors in oropharyngeal mucositis is not fully understood. Therefore, we conducted this study to determine possible associations of age, sex, underlying disease, type of chemotherapy and ABO blood group antigens with the risk of chemotherapy-induced oropharyngeal mucositis. METHODS: A total of 641 patients (395 boys and 246 girls; mean age 6.82+/-4.08 years) treated by standard chemotherapy for different type of malignancies were enrolled in the study. Mucositis was scored using the WHO scale. RESULTS: Oropharyngeal mucositis was found in 65.4% of our population. Patients with hematological malignancies (RR=1.87; 95% CI 1.33-2.67; P<0.0001) and under antimetabolities drugs (RR=1.88; 95% CI 1.33-2.63; P<0.0001) were associated with increased risk of oropharyngeal mucositis. Also, patients with blood group O were at higher risk (RR=2.86; 95% CI 2.03-4.02; P<0.0001) compared to patients with blood type A (RR= 0.47; 95% CI 0.33-0.66; P<0.0001) and blood type B (RR=0.59; 95% CI 0.38-0.91; P= 0.01). No relationship was found between oropharyngeal mucositis and age or sex. CONCLUSIONS: To our knowledge this is the first report demonstrating an association between ABO blood group and oropharyngeal mucositis. Further investigations are needed for a better understanding of this relationship.

The diversity, dynamics and activity of Castelmagno PDO cheese microbiota were studied in three batches produced in a floor valley farm, in the Grana Valley (northwest Italy), during the wintertime. Samples of milk, curd and cheese (core and subsurface) at different ripening time were submitted to both culture-dependent and -independent analysis. In particular, DNA and RNA directly extracted from the matrices were studied by PCR-Denaturing gradient gel electrophoresis (DGGE) and reverse transcription (RT)-PCR-DGGE. Culture-dependent methods highlighted the initial dominance of a thermophilic streptococcal population with the species Streptococcus thermophilus and S. agalactiae. Then, mesophilic lactococci occurred among isolates during manufacturing, with Lactococcus lactis which was also well represented in the first month of Castelmagno PDO ripening. At this point and throughout the ripening, lactobacilli prevailed in cheese samples, represented from Lactobacillus plantarum and Lb. casei. Culture-independent analysis underlined the undoubted role of L. lactis, actively involved in both Castelmagno PDO manufacturing and ripening. Despite Lb. helveticus was never isolated on selective media, a DGGE band referred to this microorganism was detected, at RNA level, in samples from ripened cheeses. On the other hand, Lb. plantarum was widely isolated from the plates, among lactobacilli, but never detected by direct analysis. Due to the importance of microbiota in the sensory richness and properties of traditional cheeses, new information have been added, in this work, on microbial diversity of Castelmagno PDO cheese.

Natural populations of lactic acid bacteria (LAB) and silage fermentation of vegetable residues were studied. Fifty-two strains of LAB isolated from cabbage, Chinese cabbage, and lettuce residues were identified and characterized. The LAB strains were gram-positive and catalase-negative bacteria, which were divided into 6 groups (A to F) according to morphological and biochemical characteristics. The strains in group A were rods that did not produce gas from glucose and formed the d and l isomers of lactate. Groups B and C were homofermentative cocci that formed l-lactic acid. Groups D, E, and F were heterofermentative cocci that formed d-lactic acid. Based on 16S rDNA gene sequence analysis, group A to F strains were identified as Lactobacillus plantarum, Lactococcus piscium, Lactococcus lactis, Leuconostoc citreum, Weissella soli and Leuconostoc gelidum, respectively. The prevalent LAB, predominantly homofermentative lactobacilli, consisted of Lactobacillus plantarum (34.6%), Weissella soli (19.2%), Leuconostoc gelidum (15.4%), Leuconostoc citreum (13.5%), Lactococcus lactis (9.6%), and Lactococcus piscium (7.7%). Lactobacillus plantarum was the dominant member of the LAB population in 3 types of vegetable residues. These vegetable residues contained a high level of crude protein (20.2 to 28.4% of dry matter). These silages prepared by using a small-scale fermentation system were well preserved, with low pH and a relatively high content of lactate. This study suggests that the vegetable residues contain abundant LAB species and nutrients, and that they could be well preserved by making silage, which is a potentially good vegetable protein source for livestock diets.

Sixty-four exopolysaccharide-producing thermophilic lactic acid bacteria (LAB) were isolated from traditionally made Indian fermented milk products. On the basis of morphological and biochemical tests, these isolates were identified as the species of Lactobacillus, Streptococcus, and Enterococcus genera. Initial screening for technological attributes revealed that Streptococcus thermophilus IG16 was a promising isolate, and produced both capsular and ropy polysaccharides at the concentration of 211mg/L. Exopolysaccharide produced by IG16 was a heteropolysaccharide containing rhamnose and galactose in a ratio of 5.3:1 and had a molecular weight of 3.3 x 10(4) Da. Use of IG16 as a starter culture controlled whey separation and improved viscosity, flavor, consistency, and color and appearance of lassi. Use of IG16 resulted in lassi having optimal acidity, less syneresis, high viscosity, and better scores for flavor, consistency, and color and appearance.

The microflora of four batches of traditional Greek Graviera cheese was studied at 5 weeks of ripening, and 200 lactic acid bacteria (LAB) isolates were phenotypically characterized and screened for antilisterial bacteriocins. The cheeses were also analyzed for organic acids by high-performance liquid chromatography and for the potential presence of 25 known LAB bacteriocin genes directly in cheese and their microbial consortia by PCR. All batches were safe according to the European Union regulatory criteria for Listeria monocytogenes, Salmonella, enterobacteria, and coagulase-positive staphylococci. The cheese flora was dominated by nonstarter Lactobacillus casei/paracasei (67.5%) and Lactobacillus plantarum (16.3%) strains, whereas few Streptococcus thermophilus (3.8%), Lactococcus lactis subsp. lactis (0.6%), and Leuconostoc (1.9%) organisms were present. Enterococcus faecium (9.4%) and Enterococcus durans (0.6%) were isolated among the dominant LAB from two batches; however, enterococci were present in all batches at 10- to 100-fold lower populations than mesophilic lactobacilli. Sixteen E. faecium isolates produced antilisterial enterocins. In accordance, enterocin B gene was detectable in all cheeses and enterocin P gene was present in one cheese, whereas the consortia of all cheeses contained at least two of the enterocin A, B, P, 31, L50A, and L50B genes. Plantaricin A gene was also amplified from all cheeses. Mean concentrations of lactic, acetic, citric, and propionic acids in the ripened cheeses exceeded 1.5% in total, of which approximately 0.9% was lactate. Thus, organic acid contents constitute an important hurdle factor for inhibiting growth of pathogens in traditional Graviera cheese products, with LAB bacteriocins, mainly enterocins, potentially contributing to increased cheese safety.

BACKGROUND: The main whey proteins alpha-lactalbumin (alpha-LA) and beta-lactoglobulin (beta-LG) are considered as the major allergens in cow's milk. Microbial fermentation can produce some proteolytic enzymes, which can induce the degradation of milk protein allergens. In this study, the effects of fermentation by lactic acid bacteria on the antigenicity of alpha-LA and beta-LG were investigated using indirect competitive ELISA. Meanwhile, the proteolysis of milk proteins was detected by TNBS assay and SDS-PAGE electrophoresis.RESULTS: Fermentation by lactic acid bacteria could significantly reduce the antigenicity of alpha-LA and beta-LG in skim milk. Combined strains of Lactobacillus helveticus and Streptococcus thermophilus were the most effective in reducing the antigenicity of both whey proteins. In addition, alpha-LA and beta-LG antigenicity decreased to a lower value at 6 h of fermentation and at 0.5 d of cold storage by fermentation with the combined strains. The results of TNBS assay and SDS-PAGE electrophoresis showed that lactic acid bacteria strains used in this study hydrolysed whey proteins only to a limited extent.CONCLUSION: The fermentation with lactic acid bacteria is an effective way to reduce whey proteins antigenicity. Copyright (c) 2010 Society of Chemical Industry.

Five samples of Airag and 20 of Tarag (both in Mongolia) were collected from scattered households. One hundred strains of lactic acid bacteria (LAB) were isolated and identified from these samples according to phenotypic characterization and 16S rRNA gene sequence analysis. Eighty-five isolates belonged to the genus Lactobacillus, 15 being classified as coccoid LAB. All isolates belonged to 5 genera and 11 to different species and subspecies. Lactobacillus (Lb.) helveticus was predominant population in Airag samples, Lb. fermentum and Lb. helveticus were the major LAB microflora in Tarag.

Twenty-four strains of lactic acid bacteria (LAB) isolated from a traditional Spanish cheese (Genestoso cheese) were evaluated for their enzymatic activities (acidifying and proteolytic abilities and carboxypeptidase, aminopeptidase, dipeptidase, caseinolytic and esterase activities), in order to select indigenous strains of technical interest for the manufacture of cheese. These strains were selected on the basis of their antimicrobial activity relative to five reference strains and were identified as Lactococcus lactis subsp. lactis (thirteen strains), Leuconostoc mesenteroides (two strains), Leuconostoc pseudomesenteroides (one strain), Lactobacillus paracasei (two strains), Lactobacillus plantarum (one strain) and Enterococcus faecalis (five strains). Lactococcus strains were those that showed the greatest degree of acidifying and proteolytic activity. The cell-free extracts (CFE) of L. paracasei exhibited the highest level of aminopeptidase activity. The highest level of caseinolytic activity was shown by the CFE of one strain of L. lactis. High values were also obtained with the CFE of Lactobacillus and of several Leuconostoc. The highest level of dipeptidase activity was found amongst the strains of L. lactis. Carboxypeptidase activity was generally very low or undetectable for the majority of strains. The greatest degree of esterolytic activity was detected for Enterococcus.

Abstract Aim: To study the role of beta-glucosidase producing probiotic bacteria and yeast in the biotransformation of isoflavone glycosides to aglycones, mineral bioavailability and vitamin B complex in fermented soymilk. Methods and Results: Five isolates of probiotic lactic acid bacteria (LAB), Lactobacillus acidophilus B4496, Lactobacillus bulgaricus CFR2028, Lactobacillus casei B1922, Lactobacillus plantarum B4495 and Lactobacillus fermentum B4655 with yeast Saccharomyces boulardii were used to ferment soymilk to obtain the bioactive isoflavones, genistein and daidzein. High-performance liquid chromatography was used to analyse the concentration of isoflavones. Bioactive aglycones genistein and daidzein after 24 and 48 h of fermentation ranged from 97.49 to 98.49% and 62.71 to 92.31% respectively with different combinations of LAB with yeast. Increase in bioavailability of minerals and vitamin B complex were also observed in fermented soymilk. Conclusions: LAB in combination with yeast S. boulardii has great potential for the enrichment of bioactive isoflavones, enhancing the viability of LAB strains, decreasing the antinutrient phytic acid and increasing the mineral bioavailability in soymilk fermentation. Significance and Impact of the Study: Fermentation of soymilk with probiotic organisms improves the bioavailability of isoflavones, assists in digestion of protein, provides more soluble calcium, enhances intestinal health and supports immune system. Increased isoflavone aglycone content in fermented soymilk improves the biological functionality of soymilk.

The bacteriocins produced by Lactococcus lactis subsp. lactis C101910 (C101910) and NBRC 12007 (NBRC 12007) were used to prevent the growth of sake spoiling hiochi bacteria (Lactobacillus hilgardii, Lactobacillus fructivorans, and Lactobacillus paracasei) in namazake, which is raw (unpasteurized) sake. The bacteriocin concentrations required for decreasing the viable cell concentrations of L. hilgardii and L. fructivorans below the detection limit (1.0 x 10(2) cells/ml) in 24 h from the initial concentration of 4.0-9.5 x 10(5) cells/ml in the namazake at pH 4.5 and at 4 degrees C, were 18-35 U/ml and 5.6 U/ml for the bacteriocin from C101910 and NBRC 12007, respectively. To decrease the viable cell concentration of L. paracasei from the initial concentration of 7.5 x 10(5) cells/ml to below the detection limit (1.0 x 10(2) cells/ml) in 24 h, 350 U/ml bacteriocin from C101910 and 140 U/ml bacteriocin from NBRC 12007 were required. In experiments using McIlvaine buffer (pH 4.5) with 15% ethanol instead of namazake as the medium, the viable cell concentrations of L. hilgardii and L. paracasei decreased to less than 1.0 x 10(2) cells/ml, whereas those of L. fructivorans decreased to less than 1.0 x 10(3) cells/ml, when bacteriocins were added at the concentrations that had proven effective in namazake. The membrane depolarization assay using a fluorescent probe showed that the presence of ethanol stimulated the collapse of the membrane potential induced by bacteriocins. The ethanol induced collapse of the membrane potential suggests that the application of bacteriocins at the storage stage of namazake is more beneficial than when used in other stages of the sake brewing process.

The lactic acid bacteria community in traditional goat cheese produced in three dairies in Valsesia (Piemonte, Italy) was studied at different steps of the manufacturing process. These cheeses were produced from raw milk without starter bacteria, and no protocol was followed during the manufacturing process. Three hundred thirty-two isolates were characterized and grouped by results of both morphophysiological tests and random amplification of polymorphic DNA plus PCR analysis. Bacteria were identified by partial sequencing of the 16S rRNA gene. Lactococci were the dominant lactic acid bacteria in raw milk. Their initial numbers ranged from 5 to 7 log CFU ml(-1). Their levels increased during manufacturing and decreased during ripening. The growth trend for enterococci was comparable to that of lactococci, although enterococci counts were lower. Lactococcus lactis subsp. cremoris, Lactococcus garviae, and Enterococcus faecalis were the most frequently isolated species during goat cheese manufacturing, whereas the highest numbers of Enterococcus (E. faecium, E. durans, E. gilvus, and E. casseliflavus) were isolated with the greatest frequency from ripened cheese samples. Occasionally, Leuconostoc mesenteroides, Leuconostoc lactis, and Lactobacillus paraplantarum also were isolated.