Biocontrol of Listeria monocytogenes by lactic acid bacteria isolated from brewer's grains used as feedstuff in Argentina

Brewer's grains are the most important by-product of the brewery industry and it is mostly used as a protein and energy source in animal nutrition. Listeria monocytogenes is a food-borne pathogen can cause invasive diseases such as meningoencephalitis, sepsis, abortion, and gastroenteritis in humans and several animal species. The aim of this work was to study the antilisterial activity of lactic acid bacteria (LAB) isolated from brewer's grains. The incidence of Listeria spp. in brewer's grains was 3.12%. Twenty-one LAB inhibited the growth of the eight strains of L. monocytogenes. The mean inhibition halo of cell free supernatants of LAB ranged between 11.5 and 24.5 mm. The isolation of lactic acid bacteria with antilisterial activity from brewer's grains is promising based on their capacity to produce antimicrobial compounds. The production of antimicrobial metabolites by LAB in the substrate would generate an unfavorable environment for the growth of the pathogenic bacterium under study.     

Application of essential oils in maize grain: impact on Aspergillus section Flavi growth parameters and aflatoxin accumulation

The antifungal activity of Pimpinella anisum L. (anise), Pëumus boldus Mol (boldus), Hedeoma multiflora Benth (mountain thyme), Syzygium aromaticum L. (clove), and Lippia turbinate var. integrifolia (griseb) (poleo) essential oils (EOs) against Aspergillus section Flavi was evaluated in sterile maize grain under different water activity (a_w) condition (0.982, 0.955, and 0.90). The effect of EOs added to maize grains on growth rate, lag phase, and aflatoxin B₁ (AFB₁) accumulation of Aspergillus section Flavi were evaluated at different water activity conditions. The five EOs analyzed have been shown to influence lag phase and growth rate. Their efficacy depended mainly on the essential oil concentrations and substrate water activity conditions. All EOs showed significant impact on AFB₁ accumulation. This effect was closely dependent on the water activity, concentration, and incubation periods. Important reduction of AFB₁ accumulation was observed in the majority of EO treatments at 11 days of incubation. Boldus, poleo, and mountain thyme EO completely inhibited AFB₁ at 2000 and 3000 μg g⁻¹. Inhibition of AFB₁ accumulation was also observed when aflatoxigenic isolates grew with different concentration of EOs during 35 days.     

Potential use of phenolic antioxidants on peanut to control growth and aflatoxin B1 accumulation by Aspergillus flavus and Aspergillus parasiticus

Food‐grade antioxidants: butylated hydroxyanisole (BHA), propyl paraben (PP) and butylated hydroxytoluene (BHT) (10 and 20 mmol g^?1) and all the mixtures of these chemicals were tested for inhibitory activity on the growth of and aflatoxin B₁ (AFB₁) accumulation by Aspergillus parasiticus and A. flavus on irradiated (7 kGy) peanut grains. Also, the influence of these treatments was evaluated in different water conditions (0.982, 0.955, 0.937a_w) at 11 and 35 days of incubation at 28 °C. Water activity (a_w) affected the fungal growth, no fungal development was observed at the highest stress water condition (0.937a_w). Butylated hydroxyanisole at 10 mmol g^?1 level and all the mixtures with PP and/or BHT were significantly effective (P = 0.05) in increasing lag phase and reducing growth rate and colony forming units per gram of peanut of both Aspergillus section Flavi strains and AFB₁ accumulation. The application of BHA at concentrations of 20 mmol g^?1 alone or with PP and/or BHT totally inhibited fungal growth at 11 and 35 days of incubation. The results suggest that the addition of these chemical mixtures on peanut grains at low levels has potential to impact synergically on the control of Aspergillus section Flavi. Copyright © 2007 Society of Chemical Industry     

Evaluation of food grade antioxidant formulation for sustained antifungal,antiaflatoxigenic and insecticidal activities on peanut conditioned at different water activities

The aim of this study was to investigate antifungal and insecticidal activity of two microencapsulated antioxidants: 2(3)-tert-butyl-4 hydroxyanisole (BHA) and 2,6-di(tert-butyl)-p-cresol (BHT) against Aspergillus section Flavi and Oryzaephilus surinamensis (L.), a vector carrier of aflatoxigenic fungi on stored peanuts. Susceptibility of Aspergillus section Flavi, insects, and aflatoxin B₁ accumulation in sterile peanut kernels conditioned at two different water activities (a_w) (0.83 a_w and 0.95 a_w) was determined with different doses of antioxidant formulations (10, 20 and 30 mM) during 45 days. Moreover, Aspergillus section Flavi isolation frequency from live and dead insects was evaluated. The BHA formulation completely inhibited Aspergillus section Flavi development regardless of a_w and doses assayed. Antifungal effect of microencapsulated BHT was highly dependent on a_w, with 86–100% fungal inhibition at 20 and 30 mM, at the lowest a_w (0.83 a_w) and at the end of the experiment. No aflatoxin accumulation was detected in samples treated with the BHA formulation. In general, low levels of Aspergillus section Flavi were detected in dead insects. Our results show efficacy for 45 days, in addition microencapsulated BHT could be an alternative to control peanut pests in dry kernels.     

Predominant mycobiota and aflatoxin content in Brazil nuts

The Brazil nut (Bertholletia excelsa) is an economically important product to the Brazilian Amazon. Currently, its marketing is being affected by the high incidence of aflatoxins (AF) produced by potentially aflatoxigenic fungi associated with its seeds. In this context, the purpose of this study was to determine which part of the nut contributes to contamination by aflatoxins and to identify the mycobiota in Brazil nut samples. Unshelled and shelled nuts were analyzed by measuring the total count of filamentous fungi (Aspergillus sections Flavi, Nigri and Circumdati) in sanitised and non-sanitised treatments. The isolates identified as Aspergillus section Flavi, the major producers of AF, were plated for determination of their aflatoxigenic potential. To perform the AF analysis, samples of Brazil nuts were treated separately. The AF from the shell and kernel were extracted by chloroform and analysed by the HPLC-FD system in isocratic mode. The Aspergillus section Flavi count was 21.67% lower. The production of AF by the isolated fungi was 30% for sanitised and 23.8% for non-sanitised samples. The concentrations obtained of AFB₁ and AFG₁ were higher than those of AFB₂ and AFG₂. The AFB₁ concentrations of shelled nuts and shell samples were 35.0 and 1.78???g/kg, respectively. AFB₂ and AFG₂ were detected only in shelled nut samples. The HPLC-FD analysis presented limits of detection (LOD) and quantification (LOQ) of 0.2 and 0.4???g/kg, respectively.     

A survey on distribution and toxigenicity of Aspergillus section Flavi in poultry feeds

Thirty-five samples of poultry feeds and corresponding raw materials (maize, soybean and meat meal) from a processing plant were analyzed to evaluate the distribution and toxigenicity of Aspergillus section Flavi isolates. Mycological analysis of the samples indicated the presence of five fungal genera (Aspergillus, Penicillium, Fusarium, Cladosporium, and Eurotium). Aspergillus flavus was the predominant species being present in 48.5% of the analyzed samples. Ninety-one isolates belonging to Aspergillus section Flavi were isolated; ninety were identified as A. flavus and only one as A. parasiticus. Fifty-seven isolates were capable of producing sclerotia, 41 were identified as L-type strains and 16 as type S. Fifty-seven percent of the isolates produced AFB₁ levels ranging from 0.05 μg/kg to 27.7 μg/kg whereas 86.8% produced CPA from 1.5 μg/kg to 137.8 μg/kg. L-strains produced from 0.05 to 14.8 μg/kg of aflatoxin and type S produced levels from 0.05 to 1.65 μg/kg. No significant differences in CPA production among S- and L-strains were observed. Sclerotial isolates produced AFB₁ levels ranging between 0.05 and 27.7 μg/kg and CPA levels from 3.8 to 47.3 μg/kg. More than half of the A. flavus isolates were able to produce AFB and CPA simultaneously. Twenty percent of the 35 samples were contaminated with aflatoxin B₁ whereas 34.3% were contaminated with CPA. The high rate of CPA producing isolates represents a potential risk of contamination with this toxin in poultry feeds.     

Toxigenic molds in Tunisian and Egyptian sorghum for human consumption

The objective of this study was to characterize the mycoflora of sorghum grains commercialized in the Tunisian retail market and to identify aflatoxins (AFs), ochratoxin A (OTA) and zearalenone (ZEA) producing species. Sixty four samples of sorghum (37 samples of Tunisian sorghum and 27 samples of Egyptian sorghum) were analyzed. Dilution plating (CFU, colony forming units) was used for fungal enumeration. The isolation of mycobiota was carried out by plating of grains on PDA and malachite green medium. Aspergillus section Flavi and section Nigri and Fusarium isolates were sub-cultured in CYA to test their ability to produce AFs, OTA and ZEA, respectively. The selected Aspergillus section Flavi and section Nigri, Penicillium and Fusarium isolates were subjected to specific PCR assays using published species-specific primers. The results revealed the dominance of Fusarium (95.3%), followed by Aspergillus (87.2%) and Alternaria (81.2%) species. The fungal counts ranged from 100 to 1.3·10⁴ CFU/g for Tunisian sorghum and from 100 to 5.7·10³ CFU/g for Egyptian sorghum. Among Aspergillus section Flavi isolates identified by molecular biology, Aspergillus flavus was the most dominant (90.1%) while Aspergillus parasiticus represent 9.9% only. About Aspergillus section Nigri, results showed the dominance of Aspergillus niger aggregate species, including Aspergillus niger, Aspergillus tubingensis and other species. Among Fusarium species, Fusarium incarnatum was the most dominant in both Tunisian and Egyptian sorghum. Penicillium citrinum was the dominant Penicillium species in the studied samples. More than 890 isolates belonging to the genus Aspergillus and Fusarium were tested in order to test their capacity to produce AFs, OTA and ZEA. The percentage of mycotoxin producing isolates in Aspergillus section Flavi, A. section Nigri, and Fusarium was 30.0%, 4.6% and 11.1%, respectively.     

Predominant mycobiota and aflatoxin content in Brazil nuts

The Brazil nut (Bertholletia excelsa) is an economically important product to the Brazilian Amazon. Currently, its marketing is being affected by the high incidence of aflatoxins (AF) produced by potentially aflatoxigenic fungi associated with its seeds. In this context, the purpose of this study was to determine which part of the nut contributes to contamination by aflatoxins and to identify the mycobiota in Brazil nut samples. Unshelled and shelled nuts were analyzed by measuring the total count of filamentous fungi (Aspergillus sections Flavi, Nigri and Circumdati) in sanitised and non-sanitised treatments. The isolates identified as Aspergillus section Flavi, the major producers of AF, were plated for determination of their aflatoxigenic potential. To perform the AF analysis, samples of Brazil nuts were treated separately. The AF from the shell and kernel were extracted by chloroform and analysed by the HPLC-FD system in isocratic mode. The Aspergillus section Flavi count was 21.67% lower. The production of AF by the isolated fungi was 30% for sanitised and 23.8% for non-sanitised samples. The concentrations obtained of AFB₁ and AFG₁ were higher than those of AFB₂ and AFG₂. The AFB₁ concentrations of shelled nuts and shell samples were 35.0 and 1.78 μg/kg, respectively. AFB₂ and AFG₂ were detected only in shelled nut samples. The HPLC-FD analysis presented limits of detection (LOD) and quantification (LOQ) of 0.2 and 0.4 μg/kg, respectively.     

Effect of synthetic antioxidants on stored maize grain mycoflora in situ

BACKGROUND: The influence of a mixture of butylated hydroxyanisole (BHA) and propyl paraben (PP) (each at a concentration of 20 mmol L^?1) on mycoflora and Aspergillus section Flavi populations in stored maize grain was evaluated. A survey of 120 maize samples was carried out from June to November 2005. RESULTS: The predominant populations in non‐treated (control) maize between the first and sixth sampling periods were Aspergillus section Flavi and Penicillium. Aspergillus flavus was the fungus most frequently isolated from both control and antioxidant‐treated kernels. All samples of control and antioxidant‐treated maize kernels were negative for aflatoxins during the 6 month storage period. Aspergillus flavus and A. parasiticus strains showed a variable ability to produce aflatoxins. The contribution of the strains to silo community toxigenicity was higher for A. flavus L (large) and S (small) strains in the fourth sampling period. CONCLUSION: Antioxidant treatment negatively affected natural maize mycoflora and Aspergillus section Flavi populations between the second and sixth months of storage. Copyright © 2007 Society of Chemical Industry     

Polyphasic characterization of Aspergillus section Flavi isolated from animal feeds in Algeria

In Algeria, little information is available on the population structure of Aspergillus section Flavi in raw materials and resultant animal feeds. A total of 172 isolates belonging to Aspergillus section Flavi were recovered from 57 animal feeds and identified on the basis of macro and micro-morphological characters, mycotoxin production and genetic relatedness. For the molecular analysis, sequencing of the calmodulin gene (CaM) and the internal transcribed spacer (ITS) regions were performed for representative isolates. Four distinct morphotypes were distinguished: Aspergillus flavus (78.5%), Aspergillus tamarii (19.2%), Aspergillus parasiticus (1.7%), and Aspergillus alliaceus (0.6%). All A. flavus isolates were of the L type and no correlation between sclerotia production and aflatoxigenicity was observed. Our results showed that 68% of the A. flavus strains produced aflatoxins B (AFB), and 72.7% were cyclopiazonic acid (CPA) producers. The three isolates of A. parasiticus were able to produce AFB and aflatoxins G but not CPA whereas, all the strains of A. tamarii produced only CPA. The obtained results revealed the presence of different species of Aspergillus section Flavi, among which were aflatoxin producers. This study provides evidence useful for considerations in aflatoxin control strategies.     

Impact of volatile compounds generated by essential oils on Aspergillus section Flavi growth parameters and aflatoxin accumulation

BACKGROUND: The advantage of essential oils is their bioactivity in the vapour phase, a characteristic that makes them attractive as possible fumigants for stored grain protection. In this study the antifungal and antiaflatoxigenic effects of the volatile fractions of five essential oils (EOs) were evaluated by vapour contact on Aspergillus section Flavi isolates. RESULTS: In maize meal extract agar the volatile fractions of Pimpinella anisum L. (anise), Pëumus boldus Mol. (boldus), Hedeoma multiflora Benth. (mountain thyme), Lippia turbinata var. integrifolia (Griseb.) (poleo) and Syzygium aromaticum L. (clove) were able to decrease the growth rate and lag phase of aflatoxigenic isolates. Boldus EO showed the best antifungal effect on Aspergillus section Flavi growth rate. In sterilised maize grains, boldus and poleo EOs showed antifungal effects on growth rate and aflatoxin accumulation. The volatile fraction of boldus EO completely inhibited the growth of isolates at water activity (a_w) levels of 0.955, 0.930 and 0.900, while poleo EO showed this effect only at the lower a_w levels (0.930 and 0.900). All aflatoxigenic isolates showed reduced total aflatoxin accumulation in the presence of boldus EO under all a_w conditions. CONCLUSION: These findings clearly indicate that the volatile fraction of boldus EO could be used to control aflatoxigenic fungi in stored maize. Copyright © 2009 Society of Chemical Industry     

Diversity and functional characteristics of lactic acid bacteria from traditional kefir grains

Traditional kefir grains were collected from distinct parts of Turkey, and their microbial profile was determined. A wide bacterial biota was observed formed by distinct lactic acid bacteria (LAB) in which Lactococcus lactis strains appeared to be dominant. Yeast species were also identified in kefir grains. Significant levels of antifungal and antibacterial activities were monitored in kefir isolates. All tested LAB produced an exopolysaccharide (EPS) containing glucose and galactose, and some strains formed a fructan‐type EPS. Importantly, low levels of antibiotic resistance were observed among the kefir isolates.     

Pediococcus acidolactici and Pediococcus pentosaceus isolated from a rainbow trout ecosystem have probiotic and ABF1 adsorbing/degrading abilities in vitro

ABSTRACT

Probiotics are being used in biological control of bacterial pathogens, as an alternative to antibiotics, to improve health and production parameters in fish farming. Fish farming production is severely affected by aflatoxins (AFs), which are a significant problem in aquaculture systems. Aflatoxins exert substantial impact on production, causing disease with high mortality and a gradual decline of reared fish stock quality. Some aspects of aflatoxicosis in fish, particularly its effects on the gastrointestinal tract, have not been well documented. The aim of the present study was to evaluate probiotic properties of lactic acid bacterial (LAB) strains isolated from rainbow trout intestine and feed. Moreover, AFB₁-binding and/or degrading abilities were also evaluated to assess their use in the formulation of feed additives. Growth at pH 2, the ability to co-aggregate with bacterial pathogens, inhibition of bacterial pathogens, and determination of the inhibitory mechanism were tested. Aflatoxin B₁ (AFB₁) adsorption and degradation ability were also tested. All strains were able to maintain viable (10⁷ cells ml^–1) at pH 2. Pediococcus acidilactici RC001 and RC008 showed the strongest antimicrobial activity, inhibiting all the pathogens tested. The strains produced antimicrobial compounds of different nature, being affected by different treatments (catalase, NaOH and heating), which indicated that they could be H₂O_2, organic acids or proteins. All LAB strains tested showed the ability to coaggregate pathogenic bacteria, showing inhibition percentages above 40%. Pediococcus acidilactici RC003 was the one with the highest adsorption capacity and all LAB strains were able to degrade AFB₁ with percentages higher than 15%, showing significant differences with respect to the control. The ability of some of the LAB strains isolated in the present work to compete with pathogens, together with stability against bile and gastric pH, reduction of bioavailability and degradation of AFB₁, may indicate the potential of LAB for use in rainbow trout culture.     

Increase in aflatoxins due to Aspergillus section Flavi multiplication during the aerobic deterioration of corn silage treated with different bacteria inocula

The growth of Aspergillus flavus and the production of aflatoxins (AF) during the aerobic deterioration of corn silage represent a problem for animal and human health. This experiment was conducted to evaluate whether the presence of A. flavus and AF production originate from the field or additional AF are produced during the fermentation phase or during aerobic deterioration of corn silage. The trial was carried out in northern Italy on corn at a dry matter (DM) level of 34%. The fresh herbage was either not treated (C) or treated with a Lactobacillus buchneri (LB) NCIMB 40788 [(at 3 × 10⁵ cfu/g of fresh matter (FM)], Lactobacillus hilgardii (LH) CNCM I-4785 (at 3 × 10⁵ cfu/g of FM), or their combination (LB+LH; at 1.5 × 10⁵ cfu/g of FM of each strain) ensiled in 20-L silos and opened after 250 d of ensiling. After silo opening, the aerobic stability was evaluated and samples were taken after 7 and 14 d of air exposure. The pre-ensiled material, the silages at silo opening, and the aerobically exposed silages were analyzed for DM content, fermentative profiles, microbial count, nutritive characteristics, DM losses, and AFB₁, AFB₂, AFG₁, and AFG₂ contents. Furthermore, a subsample of colonies with macromorphological features of Aspergillus section Flavi was selected for AF gene pattern characterization and in vitro AF production. The presence of A. flavus was below the detection limit (<1.00 log₁₀ cfu/g) in the fresh forage before ensiling, whereas it was found in 1 out of 16 silage samples at silo opening at a level of 1.24 log₁₀ cfu/g. The AF were found in both the fresh forage and at opening in all the samples, with a predominance of AFB₂ (mean value of 1.71 μg/kg of DM). The inoculation of lactic acid bacteria determined a reduction in the lactic-to-acetic ratio compared with the control. A larger amount of acetic acid resulted in a lower yeast count and higher aerobic stability in the treated silages than in the control ones. At the beginning of aerobic deterioration, the yeasts increased to over 5 log₁₀ cfu/g, whereas the molds were close to the value observed at silo opening. When the inhibiting conditions were depleted (pH and temperature higher than 5 and 35°C, respectively), both the total molds and A. flavus reached higher values than 8.00 and 4.00 log₁₀ cfu/g, respectively, thus determining the ex novo production of AFB₁ during aerobic deterioration, regardless of treatments. The analysis of gene pattern showed that 64% of the selected colonies of A. flavus showed the presence of all 4 AF gene patterns, and 43% of the selected colonies were able to produce AF in vitro. During air exposure, after 1,000°C·h have been cumulated, starch content decreased (below 10% DM) and concentration of neutral detergent fiber, acid detergent fiber, hemicelluloses, crude protein, and ash increased. The inoculation with LB and LB+LH increased the aerobic stability of the silages and delayed the onset of aerobic microbial degradation, which in turn indirectly reduced the risk of A. flavus outgrowth and AFB₁ production after silage opening.     

西藏高原粮油作物曲霉菌污染及菌株产毒力研究

为探明西藏高原粮油作物曲霉菌污染状况及黄曲霉菌产毒能力,连续5年对西藏青稞、小麦、花生3种作物中曲霉菌污染情况进行分析,并对其分离到的黄曲霉菌株开展产毒力研究,结果表明,204份样品中,共分离出15种曲霉菌,曲霉菌污染率呈花生>青稞>小麦。青稞、小麦中曲霉属优势种均为黑曲霉(Aspergillus niger),真菌毒素主要为杂色曲霉毒素和赭曲霉毒素;花生优势种为黄曲霉(A.flavus);仅受黄曲霉毒素污染。来源于不同作物的黄曲霉菌,其产毒类型也有差异,麦类作物产毒菌株以产黄曲霉毒素B₁(AFB₁)、黄曲霉毒素B₂(AFB₂)为主;花生产毒菌株以产AFB₁、AFB₂、AFG₁、AFG₂为主。     

In vitro activity of the Brazil nut (Bertholletia excelsa H.B.K.) oil in aflatoxigenic strains of Aspergillus parasiticus

The growing demand for safe foods without preservatives has fostered research to investigate the effects of natural compounds on microorganisms. Accordingly, an in vitro study was carried out to determine physico-chemical characterization and investigate the antimicrobial effects of Brazil nut oil on toxigenic strains of Aspergillus. The antifungal activity testing was carried out isolating A. parasiticus by transferring the strains to the center of the plates containing treatments with different concentrations of Brazil nut oil. The plates were incubated for 8 days at 28 °C with subsequent extraction of toxins. The aflatoxins AFB₁ AFB₂, AFG₁, and AFG₂ were determined by thin-layer chromatography. The values found for acidity, refractive index, and density ranged from 0.3512 to 0.7092, 1.4670 to 1.470, and 9.2 × 10² to 16.7 × 10², respectively. The results obtained showed that the effect of Brazil nut oil on the growth diameter of the fungal colony was time and concentration dependent. After 8 days of incubation, all treatments showed better fungal growth inhibition compared with that of the control. Total inhibition of aflatoxin production by the strain (LOQ of the method: 2 g/kg) was also observed in the medium containing the oil, while in the control treatment, AFB₁, AFB₂, and AFG₁ were produced by the inoculated strain A. parasiticus.     

Distribution of fungi and aflatoxins in a stored peanut variety

The objective of the present study was to evaluate the mycoflora and occurrence of aflatoxins in stored peanut samples (hulls and kernels) from Tupã, State of São Paulo, Brazil. The samples were analyzed monthly over a period of one year. The results showed a predominance of Fusarium spp. (67.7% in hulls and 25.8% in kernels) and Aspergillus spp. (10.3% in hulls and 21.8% in kernels), and the presence of five other genera. The growth of Aspergillus flavus was mainly influenced by temperature and relative humidity. Analysis of hulls showed that 6.7% of the samples were contaminated with AFB₁ (mean levels = 15–23.9 μg/kg) and AFB₂ (mean levels = 3.3–5.6 μg/kg); in kernels, 33.3% of the samples were contaminated with AFB₁ (mean levels = 7.0–116 μg/kg) and 28.3% were contaminated with AFB₂ (mean levels = 3.3–45.5 μg/kg). Analysis of the toxigenic potential revealed that 93.8% of the A. flavus strains isolated were producers of AFB₁ and AFB₂.     

Preliminary characterization of bacteriocin-like substances from lactic acid bacteria isolated from organic leafy vegetables

This study presents the characterization of new strains of lactic acid bacteria (LAB) from organic vegetables. Forty-five strains of LAB isolated from vegetables were investigated by its antimicrobial activity against taxonomically related microorganisms. Genetic identification of selected LAB was performed by means of PCR method. These strains were Enterococcus faecium, Lactococcus lactis, Enterococcus hirae and Enterococcus canis. Bacteriocin-like substances were active against Gram-positive bacteria and Gram-negative foodborne pathogens (Listeria monocytogenes and Escherichia coli, respectively). The antimicrobial activity of LAB strains was inactivated by the addition of proteases, thus confirming the proteinaceous nature of the inhibition. In all four strains the bacteriocin activity was stable after extended refrigerated storage and freezing-thawing cycles. This fact suggests that bacteriocin produced by the four LAB strains may find application as biopreservatives in minimally processed vegetables.     

An investigation of the bacteriocinogenic potential of lactic acid bacteria associated with wheat (Triticum durum) kernels and non-conventional flours

One hundred and thirty-seven lactic acid bacteria (LAB), previously isolated from wheat (Triticum durum) grains and non-conventional flour samples, were tested for the production of antibacterial substances. A total of 16 strains (5 Enterococcus faecium, 5 Enterococcus mundtii, 4 Pediococcus pentosaceus, 1 Lactobacillus coryniformis and 1 Lactococcus garvieae) were found to inhibit the growth of Listeria innocua. The antibacterial activities were preliminarily investigated for their general behaviour with proteolytic (proteinase K, protease B and trypsin), amylolytic (α-amylase) and lipolytic (lipase) enzymes, after heat treatment, and exposure to different pHs and ethanol concentrations. Bacteriocin-like inhibitory substances (BLIS) were also characterized for their inhibition spectra against non-pathogenic and pathogenic food-associated and human pathogenic bacteria. LAB showing the best characteristics in terms of inhibition spectrum, inhibition activity and mode of action (bactericidal) belonged to the species Ent. mundtii. The high percentage (11.68%) of BLIS-producing strains detected confirmed previous observations that raw materials may harbour higher numbers of bacteriocinogenic LAB than fermented foods.