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Propionic acid bacteria (PAB) is an umbrella term for a group of bacteria with the ability to produce propionic acid. In the past, due to this common feature and other phenotypic similarities, genetically heterogeneous bacteria were considered as a single genus, Propionibacterium. Members of this genus ranged from "dairy propionibacteria," which are widely known for their role in eye and flavor formation in cheese production, to "cutaneous propionibacteria," which are primarily associated with human skin. In 2016, the introduction of two new genera based on genotypic data facilitated a clear separation of cutaneous (Cutibacterium spp.) from dairy PAB (Propionibacterium spp., Acidipropionibacterium spp.). In light of these taxonomic changes, but with particular emphasis on dairy PAB, this review describes the current state of knowledge about metabolic pathways and other characteristics such as antibiotic resistance and virulence factors. In addition, the relevance of dairy PAB for the food industry and cheese production in particular is highlighted. Furthermore, methods for cultivation, detection, and enumeration are reviewed, incorporating the current taxonomy as well as the potential for routine applications.

As bread is a very important staple food, its spoilage threatens global food security. Ropy bread spoilage manifests in sticky and stringy degradation of the crumb, slime formation, discoloration, and an odor reminiscent of rotting fruit. Increasing consumer demand for preservative-free products and global warming may increase the occurrence of ropy spoilage. Bacillus amyloliquefaciens, B. subtilis, B. licheniformis, the B. cereus group, B. pumilus, B. sonorensis, Cytobacillus firmus, Niallia circulans, Paenibacillus polymyxa, and Priestia megaterium were reported to cause ropiness in bread. Process hygiene does not prevent ropy spoilage, as contamination of flour with these Bacillus species is unavoidable due to their occurrence as a part of the endophytic commensal microbiota of wheat and the formation of heat-stable endospores that are not inactivated during processing, baking, or storage. To date, the underlying mechanisms behind ropy bread spoilage remain unclear, high-throughput screening tools to identify rope-forming bacteria are missing, and only a limited number of strategies to reduce rope spoilage were described. This review provides a current overview on (i) routes of entry of Bacillus endospores into bread, (ii) bacterial species implicated in rope spoilage, (iii) factors influencing rope development, and (iv) methods used to assess bacterial rope-forming potential. Finally, we pinpoint key gaps in knowledge and related challenges, as well as future research questions.

The importance of microorganisms in the beet sugar industry came up in 1930. Since then, several approaches have been made to describe these bacteria. For this purpose, mainly cultivation-based methods were applied. However, the majority of the microorganisms cannot be cultivated or are in the viable-but-non-culturable state. In addition, these methods are time-consuming and costly. Progress in molecular biology allows a cheaper, faster and more precise identification of the microbiota. This study evaluates the application of an 16S rDNA-based metagenomic sequencing approach based on Illumina MiSeq technology to identify the microbiota in raw juice and juice of mid-tower in a beet sugar production plant and compares the results with those obtained by cultivation-based techniques. All bacteria orders detected with cultivation-based methods could be also found with the applied metagenomic approach. In raw juice, mainly mesophilic bacteria such as Lactobacillus spp. and Leuconostoc spp. species were identified. Additionally, a partly large proportion of gram-negative bacteria belonging to the order Enterobacterales were detected by the metagenomic approach. The diversity in juice of mid-tower was much lower and predominated by mainly thermophilic genera such as Geobacillus, Caldanaerobius and Thermoanaerobacter. The last two mentioned genera belong to the class of Clostridia. Surprisingly, in the juice of mid-tower Lactobacillus species could be verified by cultivation-based methods as well as by the metagenomic approach. As a consequence, it can be presumed that lactobacilli can survive in this very specific environment at 70 °C occurring in the central part of the extraction tower.

Insects have the potential to become an efficient and reliable food source for humans in the future and could contribute to solving problems with the current food chain. Analytical methods to verify the authenticity of foods are essential for consumer acceptance. We present a DNA metabarcoding method that enables the identification and differentiation of insects in food. The method, developed on Illumina platforms, is targeting a 200 bp mitochondrial 16S rDNA fragment, which we found to be suitable for distinguishing more than 1000 insect species. We designed a novel universal primer pair for a singleplex PCR assay. Individual DNA extracts from reference samples, DNA extracts from model foods and food products commercially available were investigated. In all of the samples investigated, the insect species were correctly identified. The developed DNA metabarcoding method has a high potential to identify and differentiate insect DNA in the context of food authentication in routine analysis.

This study aimed to simulate the impact of an African swine fever (ASF) outbreak in Austria. ASF is one of the most significant and critical diseases for the global domestic pig population. Hence, the authors evaluated control strategies and identified bottlenecks during an ASF outbreak. A hybrid approach was selected, including discrete-event and agent-based simulation. An extended Susceptible-Exposed-Infectious-Recovered (SEIR) model (within a pig farm) and a standard SEIR model (between pig farms) were used to simulate the chain of infection. A total of 576 scenarios with several parameter variations were calculated to identify the influence of external factors on key performance indicators. The main results show a comparison between two control strategies anchored in law: a standard strategy (SS) and a preventive culling strategy (SC). The calculated scenarios show a difference between these strategies and indicate that with SC during an outbreak, fewer farms would be infected, and fewer pigs would be culled. Furthermore, specific geographical areas were identified, which—due to their density of pigs and farms—would be severely affected in case of an ASF outbreak. The analysis of bottlenecks in rendering plants (RPs) showed an increase in the number of days RPs were overutilized as the transmission rate increased. In addition, SS caused more days of overutilized RPs than SC.

BACKGROUND During the manufacture of sucrose from sugar beet, different microorganisms originating from the plant material as well as from the soil enter the process. Due to the formation of polysaccharide-based slimes, these contaminants may induce several adverse effects such as filtration problems during juice purification. Certain microorganisms also metabolize sucrose, leading to product losses with financial consequences. To better understand and to prevent these negative effects, the aim of the study was to investigate the evolution of relevant bacterial groups, including their metabolites appearing during the extraction process. For this purpose, one production cycle was monitored to identify the major contamination steps and to clarify how they relate to the processing conditions. Traditionally, different antimicrobial agents such as formaldehyde, sulfur dioxide, hypochlorous acid, sodium hypochlorite, and chlorine dioxide have been added to inhibit microbial growth. In the present study, a rosin-based product derived from pine trees was applied as an alternative to those substances. RESULTS Press water, raw juice, and mid-tower juice were identified as being highly contaminated with bacteria, and processing conditions such as time, temperature and pH level significantly influenced bacterial levels and the corresponding metabolites. Among the contaminants identified, lactic acid bacteria, and mesophilic and thermophilic aerobic bacteria played a dominant role, whereas lactic acid, acetic acid, butyric acid, and ethanol were identified as typical metabolites. CONCLUSION Bacterial growth during production could be reduced by shock dosing of the rosin-based material in the extraction area. © 2020 Society of Chemical Industry.

Wheat bran (WB) is an important side product of the milling industry and can serve as dietary fiber compound for monogastric animals. The aim of this study was to evaluate the influence of native or fermented WB on the gut physiology and microbiology of laying hens. To accomplish this, 24 laying hens were fed the following diets: conventional diet without WB; 15% native WB in the diet; 15% WB fermented with Pleurotus eryngii; and 15% WB fermented with P. eryngii and a lactic acid bacterial culture. Immediately after slaughtering, digesta samples were taken from the jejunum, ileum and cecum, respectively. Total DNA was extracted and subsequently investigated with 16S DNA amplicon sequencing. Neither native nor fermented WB supplementations negatively affected the feed conversion ratio, laying performance or the relative abundances and alpha-diversity of microbiota in the intestine. Effects of WB-based diets on gut morphology were only recognized in the jejunum (reduced villum height and mucosa thickness). Likewise, WB supplementation decreased the digestibility of DM and starch. Based on these findings, it was demonstrated that different WB variants are applicable without exerting practically negative consequences on performance or on gut microbiota. Fermentation improved the digestibility/retention of dietary fat and phosphorus. However, no further beneficial effects were observed. This study also allowed a more in-depth view on the laying hens' gut microbiome and its variation within the gut segments.

Clostridium tyrobutyricum is recognized as the main causative agent of late blowing defect—severe spoilage of hard and semihard cheeses. In this work, we present the draft genome sequences of 12 C. tyrobutyricum strains isolated from raw milk and cheese.

The aim of this study was to evaluate the effect of ampicillin, an organic acid-based feed additive and a synbiotic preparation on the prevalence of antibiotic-resistant E. coli in the ceca of broilers. A total of 2000 broiler chickens (Ross 708) were randomly assigned to 5 groups with 8 replicates. The negative control group was the only group that was not subjected to avian pathogenic E. coli challenge, while all the other 4 groups received a multi-resistant E. coli strain that was resistant to ampicillin, cephalexin, and nalidixic acid as an oral challenge. The second group served as a challenge control, and the third group received the antibiotic ampicillin via water for 5 d. The fourth group received a feed additive based on organic acids and cinnamaldehyde, and the fifth group received a synbiotic preparation via feed and water. On day 17 and 38 of the trial, cecal samples from 3 birds from each of the 40 pens were obtained, and the E. coli counts and abundances of antibiotic-resistant E. coli were determined. Oral challenge with an avian pathogenic E. coli strain did not influence the performance, and there was no significant difference in growth performance between groups. The total E. coli count was lower (P < 0.05) in the group supplemented with the synbiotic than in the challenge control group on day 38 of the trial. Administration of an antibiotic for 5 d led to a significant increase in the abundance of E. coli strains resistant to ampicillin, amoxicillin-clavulanic acid, cefoxitin, and ceftriaxone. There was no increase in the abundance of antibiotic-resistant E. coli observed in the groups that received feed supplemented with an organic acid/cinnamaldehyde-based feed additive or a synbiotic. Moreover, the effects of the tested feed additives on the prevalence of resistant E. coli are demonstrated by the lower ceftriaxone minimal inhibitory concentration values for this group than for the antibiotic group. Additionally, the synbiotic group exhibited lower ceftriaxone minimal inhibitory concentration values than the antibiotic group.

Cronobacter spp. are opportunistic human pathogens that cause serious diseases in neonates and immunocompromised people. Owing to their biofilm formation on various surfaces, both their detection and their removal from production plants constitute a major challenge. In this study, food samples were randomly collected in Austria and examined for the presence of Cronobacter spp. Presumptive isolates were identified by a polyphasic approach. Five percent of the samples were positive for C. sakazakii and 2.4% for C. dublinensis. Individual growth of the isolates was characterized based on lag time, growth rate, and generation time. During an incubation period of 6 to 72 h, biofilm formation of 11 selected isolates was quantified under model conditions by a crystal violet staining assay with 96-well plates with different carbon sources (lactose, glucose, maltose, sucrose, and sodium acetate) and NaCl levels and under variable temperature and pH conditions. Biofilm formation was more pronounced at lactose concentrations between 0.25 and 3% compared with 5% lactose, which lead to thinner layers. C. sakazakii isolate C7, isolated from infant milk powder, was the strongest biofilm producer at 10 mM Mg2+ and 5 mM Mn2+, 0.5% sodium acetate, at pH levels between 7 and 9 at 37°C for 24 h. C. sakazakii strain C6 isolated from a plant air filter was identified as a moderate biofilm former and C. sakazakii strain DSM 4485, a clinical isolate, as a weak biofilm former. Based on PCR detection, genes bcsA, bcsB, and bcsG encoding for cellulose could be identified as markers for biofilm formation. Isolates carrying bcsA and bcsB showed significantly stronger biofilm formation than isolates without these genes ( P < 0.05), in strong correlation with the results obtained in the crystal violet assay. Further investigations using confocal laser scanning microscopy revealed that extracellular polymeric substances and glycocalyx secretions were the dominating components of the biofilms and that the viable fraction of bacteria in the biofilm decreased over time.

In this study, the influence of meat batter composition and sausage diameter on the development of microbiota and sensory traits of traditional, spontaneously fermented wild boar meat sausages are evaluated. This research also demonstrates how principal component analysis (PCA) can be used to relate product sensory properties to particular microbial genotype and to select potential starter or adjunct culture. Generally, similar microbiological results were obtained in all types of products. The undesirable microbiota was either not detected at any sausage production stage or its number decreased below the detection limit in ripened sausages. The low growth rate of lactic acid bacteria (LAB) was consistent with the obtained pH and slow acidification rate. Although no differences in the composition of LAB species were noticed between sausage types (50S=50 % wild boar meat in small casing, 50L=50 % wild boar meat in large casing, 100S=100 % wild boar meat in small casing), a clear separation based on LAB genotypes could be observed. Upon quantitative descriptive analysis, significant differences in sensory attributes between sausage types were established. According to the PCA, the overall acceptability traits of sausages are closely linked to one Leuconostoc mesenteroides genotype (LM_4). Of all tested technological properties, LM_4 strains showed remarkable acidification ability, lowering the pH from pH=5.41 to 3.74, and pronounced proteolytic activity on skimmed milk as well as antagonistic activity against Staphylococcus aureus (DSM 20231) and Brochothrix thermosphacta (LMG 17208). Lipolytic and haemolytic activities were not detected, and all analyzed strains were susceptible to tested antibiotics and possessed no biogenic amine genes.

Food fermentation technologies have developed through years of experience rather than scientific findings. Therefore, many small-scale manufacturers are unwilling to accept changes and to modify fermentation processes. Still, traditional fermented foods have had an essential role in human nutrition for thousands of years. A wide range of diverse cereals, legumes, vegetables, fruits, fish, seafood, and meat are used as raw materials for the production of these fermented foods by back-slopping (a process where material from a previous successful batch is added to facilitate the initiation of a new batch) or naturally occurring microorganisms. Various microorganisms that influence the quality, safety, sensory properties, acceptability, and consistency of these products are present. Lactic acid bacteria with their beneficial characteristics are generally a significant contributor. Identification and profiling of microorganisms in fermented foods are of particular interest. But so far such information has not been explored in detail for some fermented products from the lower Mekong subregion. Improving the safety, quality, and acceptability of fermented foods, while reducing their production costs and maintaining their authenticity and uniqueness, is important. This can be achieved through starter culture application and greater attention to food safety, such as HACCP approaches. Recent scientific advances have led to the development of defined microbial starters, leading to the transfer of artisanal food production to more controlled and industrialized fermentation. The aim of this review is to determine the gaps in the knowledge of traditional fermented foods of the lower Mekong subregion and to present current information on the diversity of involved microorganisms. These data will be helpful for defining future strategies toward the sustainable production of safe fermented foods particularly those from southeast Asia.

In the present study, we isolated and screened thirty strains of GABA (&gamma, aminobutyric acid)-producing lactic acid bacteria (LAB) from traditional Indonesian fermented foods. Two strains were able to convert monosodium glutamate (MSG) to GABA after 24 h of cultivation at 37 &deg, C based on thin layer chromatography (TLC) screening. Proteomic identification and 16S rDNA sequencing using MALDI-TOF MS identified the strain as Lactobacillus plantarum designated as L. plantarum FNCC 260 and FNCC 343. The highest yield of GABA production obtained from the fermentation of L. plantarum FNCC 260 was 809.2 mg/L of culture medium after 60 h of cultivation. The supplementation of 0.6 mM pyridoxal 5&rsquo, phosphate (PLP) and 0.1 mM pyridoxine led to the increase in GABA production to 945.3 mg/L and 969.5 mg/L, respectively. The highest GABA production of 1226.5 mg/L of the culture medium was obtained with 100 mM initial concentration of MSG added in the cultivation medium. The open reading frame (ORF) of 1410 bp of the gadB gene from L. plantarum FNCC 260 encodes 469 amino acids with a calculated molecular mass of 53.57 kDa. The production of GABA via enzymatic conversion of monosodium glutamate (MSG) using purified recombinant glutamate decarboxylase (GAD) from L. plantarum FNCC 260 expressed in Escherichia coli was found to be more efficient (5-fold higher within 6 h) than the production obtained from fermentation. L. plantarum FNCC 260 could be of interest for the synthesis of GABA.

In the present study, we isolated and screened thirty strains of GABA-producing lactic acid bacteria (LAB) from Indonesian traditional fermented foods. Two strains were able to convert monosodium glutamate (MSG) to GABA after 24 h of cultivation at 37oC based on thin layer chromatography (TLC) screening. 16S rDNA sequencing and proteomic identification using MALDI-TOF MS identified these two strains as Lactobacillus plantarum designated as L. plantarum FNCC 260 and L. plantarum FNCC 343. The highest yield of GABA production obtained from the fermentation of L. plantarum FNCC 260 was 809.2 mg/l of culture medium after 60 h of cultivation. Supplementation of 0.6 mM pyridoxal 5’-phosphate (PLP) and 0.1 mM pyridoxine led to the increase in GABA production to 945.3 mg/l and 969.5 mg/l, respectively. The highest GABA production of 1226.5 mg/l of culture medium was obtained with 100 mM initial concentration of MSG added in the cultivation medium. The open reading frame (ORF) of 1410 bp of the gadB gene from L. plantarum FNCC 260 encodes 469 amino acids with a calculated molecular mass of 53.57 kDa. The production of GABA via enzymatic conversion of monosodium glutamate (MSG) using purified recombinant glutamate decarboxylase (GAD) from L. plantarum FNCC 260 expressed in Escherichia coli was found to be more efficient (5-fold higher within 6 h) than the production obtained from fermentation. L. plantarum FNCC 260 could be of interest for the synthesis of GABA.

Clostridium tyrobutyricum, a Gram-positive, anaerobic, spore-forming bacterium, is considered as one of the main causative agents for spoilage of hard and semihard cheeses. Growth of C. tyrobutyricum in cheese is critically influenced by ripening temperature and time, pH, salt and lactic acid concentration, moisture and fat content, and the presence of other microorganisms. Previous studies revealed high intraspecies diversity of C. tyrobutyricum strains and variable tolerance toward pH, temperatures, and salt concentrations. These findings indicate that strain-dependent characteristics may be relevant to assess the risk for cheese spoilage if clostridial contamination occurs. In this study, we aimed to compare the phenotypes of 12 C. tyrobutyricum strains which were selected from 157 strains on the basis of genotypic and proteotypic variability. The phenotypic analysis comprised the assessment of gas production and organic acid concentrations in an experimental cheese broth incubated at different temperatures (37, 20, and 14 &deg, C). For all tested strains, delayed gas production at lower incubation temperatures and a strong correlation between gas production and the change in organic acid concentrations were observed. However, considering the time until gas production was visible at different incubation temperatures, a high degree of heterogeneity was found among the tested strains. In addition, variation among replicates of the same strain and differences due to different inoculum levels became evident. This study shows, that, among other factors, strain-specific germination and growth characteristics should be considered to evaluate the risk of cheese spoilage by C. tyrobutyricum.

Preventing food spoilage without the addition of chemical food additives, while increasing functional properties of wheat-based bakery products, is an increasing demand by the consumers and a challenge for the food industry. Within this study, lactic acid bacteria (LAB) isolated from sourdough were screened in vitro for the ability to utilize the typical wheat carbohydrates, for their antimicrobial and functional properties. The dual culture overlay assay revealed varying levels of inhibition against the examined fungi, with Lactiplantibacillus plantarum S4.2 and Lentilactobacillusparabuchneri S2.9 exhibiting the highest suppression against the indicator strains Fusarium graminearum MUCL43764, Aspergillus fumigatus, A. flavus MUCL11945, A. brasiliensis DSM1988, and Penicillium roqueforti DSM1079. Furthermore, the antifungal activity was shown to be attributed mainly to the activity of acids produced by LAB. The antibacillus activity was evaluated by the spot-on-the-lawn method revealing a high inhibition potential of the majority of LAB isolated from sourdough against Bacillus cereus DSM31, B. licheniformis DSM13, B. subtilis LMG7135, and B. subtilis S15.20. Furthermore, evaluating the presence of the glutamate decarboxylase gen in LAB isolates by means of PCR showed a strain dependency of a potential GABA production. Finally, due to improved functional activities, LAB isolated from sourdoughs exhibit promising characteristics for the application as natural preservatives in wheat-based bakery products.

The quantification of nitrate and nitrite in molasses is an emerging requirement for the sugar industry. The establishment of a validated analytical methodology is necessary to comply with the EU-guidelines on maximum permissible levels currently under discussion. In the present study, validation of an anion exchange chromatographic method to determine nitrate and nitrite in molasses was performed determining key validation parameters via several stages of an interlaboratory comparison for the first time. Since a stable, homogeneous sample material was necessary for establishing quantitative accuracy, a nitrite-spiked molasses material was prepared and evaluated independently across two laboratories in a parallel study. This preliminary study revealed that quantification of nitrate and nitrite content in molasses could be achieved with excellent precision under repeatability conditions of measurement (1.21% RSD for nitrate; 1.26% RSD for nitrite) in independent laboratories but requires preservation of samples at −20 °C for long-term (>several days) storage prior the analysis. A subsequent stage involving five different participating laboratories was then undertaken to evaluate anion exchange chromatography with conductivity detection for this application within a round robin study. The first phase focused on the analysis of nitrite-spiked molasses to optimize the individual analytical practices of laboratories and to elaborate a harmonized sample preparation protocol. In the second phase the participating laboratories analyzed seven molasses samples with different pH-value, dry substance, factory origin, with nitrate levels ranging from 2931 mg/kg – 6436 mg/kg and nitrite levels ranging from 25.3 mg/kg – 109 mg/kg. The determination of precision of reproducibility conditions of measurement revealed average relative interlaboratory standard deviations of 2.5% for nitrate and of 13% for nitrite. The present study clearly demonstrates the necessity of harmonization and standardization regarding measurement and data evaluation procedures applied for controlling nitrate and nitrite in molasses.

Short-chain cello-oligosaccharides (COS; degree of polymerization, DP≤6) are promising water-soluble dietary fibers. An efficient approach to their bottom-up synthesis is from sucrose and glucose using glycoside phosphorylases. Here, we show intensification and scale up (20 mL; gram scale) of COS production to 93 g/L product and in 82 mol % yield from sucrose (0.5 M). The COS were comprised of DP 3 (33 wt %), DP 4 (34 wt %), DP 5 (24 wt %) and DP 6 (9 wt %) and involved minimum loss (≤10 mol %) to insoluble fractions. After isolation (≥95% purity; ≥90% yield), the COS were examined for growth promotion of probiotic strains. Benchmarked against inulin, trans-galacto-oligosaccharides and cellobiose, COS showed up to 4.1-fold stimulation of cell density forClostridium butyricum,Lactococcus lactissubsp.lactis,Lactobacillus paracaseisubsp.paracaseiandLb. rhamnosusbut were less efficient withBifidobacteriumsp. This study shows the COS as selectively functional carbohydrates with prebiotic potential and demonstrates their efficient enzymatic production.

Bacterial communities associated with the ripening process in artisanal wild boar and deer meat sausages were investigated by molecular barcoding using the 16S rRNA gene as a marker. A core microbiota shared by 83.54% of the samples indicated remarkable level of Lactobacillus sakei and Lactobacillus curvatus, accounting for 20.55% in initial and 70.48% in final products as well as spoilage-associated bacteria including Stenotrophomonas, Bacillus, Pseudomonas, Carnobacterium and Brochothrix, with an average abundance 44.15% at the beginning and 13.98% at the end of the production. Of selected LAB isolates (n = 555), 43.83% were not suitable for food application due to the antibiotic resistance or the presence of the tdc gene. Most of the strains designated as safe were able to grow at 25 °C even in the presence of 3.0 and 6.0% of NaCl or pH 4.5, but exposure to the same stressors resulted in growth reduction at 12 °C. Acidification and antimicrobial activity were found in 65.62% and 37.50% of strains, respectively. Most of the strains showed lipolytic and proteolytic activity, but only 9.37% were able to degrade sarcoplasmic proteins. These results give important information for the development of new starter formulation for the production of high quality game meat sausages.

This research effort aimed at isolating and phenotypically characterizing lactic acid bacteria (LAB) isolates from a spontaneous rye sourdough manufactured following traditional protocols, as well as at evaluating their antimicrobial and antifungal properties as key features for future industrial applications. Thirteen LAB strains of potential industrial interest were isolated and identified to species-level via PCR. Most of the sourdough isolates showed versatile carbohydrate metabolisms. The Leuconostoc mesenteroides No. 242 and Lactobacillus brevis No. 173 demonstrated to be gas producers, thus, revealing their heterofermenter or facultative homofermenter features. Viable counts higher than 7.0 log10 (CFU/mL) were observed for Lactobacillus paracasei No. 244, Lactobacillus casei No. 210, L. brevis No. 173, Lactobacillus farraginis No. 206, Pediococcus pentosaceus No. 183, Lactobacillus uvarum No. 245 and Lactobacillus plantarum No. 135 strains, after exposure at pH 2.5 for 2 h. Moreover, L. plantarum No. 122, L. casei No. 210, Lactobacillus curvatus No. 51, L. paracasei No. 244, and L. coryniformins No. 71 showed growth inhibition properties against all the tested fifteen pathogenic strains. Finally, all LAB isolates showed antifungal activities against Aspergillus nidulans, Penicillium funiculosum, and Fusarium poae. These results unveiled the exceptionality of spontaneous sourdough as a source of LAB with effective potential to be considered in the design of novel commercial microbial single/mixed starter cultures, intended for application in a wide range of agri-food industries, where the antimicrobial and antifungal properties are often sought and necessary. In addition, metabolites therefrom may also be considered as important functional and bioactive compounds with high potential to be employed in food and feed, as well as cosmetic and pharmaceutical applications.

Screening for various types of lactic acid bacteria (LAB) that form the biological agent &gamma, amino-butyric acid (GABA) is important to produce different kinds of GABA-containing fermented foods. So far, no GABA-producing LAB have been reported from Cambodian fermented foods. Most small-scale fermentations and even some industrial processes in this country still rely on indigenous LAB. The application of GABA-producing autochthonous starters would allow the production of Cambodian fermented foods with an additional nutritional value that meet the population&rsquo, s dietary habits and that are also more attractive for the international food market. Matrix-assisted laser desorption/ionizing time-of-flight mass spectrometry (MALDI-TOF MS) and partial 16S rDNA sequencing were used to identify 68 LAB isolates from Cambodian fermented foods. These isolates were classified and grouped with (GTG)5 rep-PCR, resulting in 50 strains. Subsequently, all strains were investigated for their ability to produce GABA by thin layer chromatography. GABA-positive strains were further analyzed by the GABase assay. Of the six GABA-positive LAB strains&mdash, one Lactobacillus futsaii, two Lactobacillus namurensis, and three Lactobacillus plantarum strains&mdash, two Lactobacillus plantarum strains produced high amounts of GABA (20.34 mM, 16.47 mM). These strains should be further investigated for their potential application as GABA-producing starter cultures in the food applications.

The deterioration of fresh food by spoilage microorganisms remains a significant problem even though a diversity of preservation approaches has been proposed. The consumer's choice is demanding for chemical and antimicrobial-free products, which has prompted the development of natural alternatives to retard food spoilage. This study was performed to evaluate the effect of the combination of essential oils (EOs) (orange, tea tree, citronella grass and cinnamon) and enterocin KT2W2G against spoilage microorganisms isolated from the surface of spoiled banana (Musa ABB cv. Kluai “Namwa”) peel by using the agar well diffusion assay. Eighteen spoilage microorganisms (9 strains of bacteria and 9 strains of yeast) were selected as indicator strains. They were identified based on partial 16S rDNA and 18S rDNA gene sequencing for bacteria and yeast, respectively. Selected spoilage bacteria were identified as Lactococcus lactis subsp. lactis, Enterococcus faecalis, Klebsiella pneumonia, Klebsiella variicola and Serratia marcescens, whereas selected spoilage yeast were identified as Kodamaea ohmeri, Pichia aff. fermentans, Pichia kudriavzevii, Hanseniaspora opuntiae and Candida metapsilosis. The agar well diffusion assays resulted in cinnamon as being the most effective EO against selected spoilage indicator. The combination of cinnamon oil and enterocin KT2W2G at 6:4 ratios exhibited the best synergistic effect against selected spoilage microorganisms, whereas enterocin KT2W2G applied as single substance displayed no activity. The results obtained indicate that the combination of cinnamon oil with enterocin KT2W2G harbours the potential to be used as a natural biocontrol agent to improve food quality and to extend the shelf life of harvested bananas.

Clostridial contamination of raw milk causes late-blowing, a severe quality defect in cheese. Consequently, milk containing high numbers of cheese-damaging clostridial spores should not be used for the production of certain types of hard and semi-hard cheese. Currently, there is no officially standardised method available to monitor clostridial spore levels in milk, and major drawbacks like long analysis time, labour intensity, uncertainty of results and insufficient selectivity for clostridia exist for usually used conventional MPN (most probable number) techniques. Therefore, an optimised medium in combination with a semi-automated application for the enumeration of clostridia in milk was developed. The aim of this study was to evaluate this new methodology in comparison with a conventional method (using Bryant and Burkey broth) based on the analysis of 84 milk samples. Method inclusivity was further tested using pure clostridial cultures, and selectivity was assessed by molecular identification of isolates obtained from the new assay. The novel approach proved to be suitable for the detection of clostridia in both suppliers’ and processed milk, also indicating that it is superior in selectivity, sensitivity and analysis time compared to conventional techniques.

The aim of this investigation was to determine the influence of seven different Lactobacillus spp. (Lb.) strains compared with a commercial starter culture (CS) on the functional properties of gluten-free (GF) sourdough-breads. The sourdough stability of selected strains was also evaluated upon back-slopping. Results showed that the bread properties were greatly affected by the Lb. strains. Millet breads achieved lower specific volumes (1.80–2.19 cm3/g), higher crumb firmness (19.01–42.19 N) and lower relative elasticities (21.5–43.4%) than buckwheat breads. Compared with the CS, Lactobacillus pentosus and Lb. hammesii positively influenced the crumb firmness of buckwheat and millet breads, respectively, while Lb. paralimentarius enhanced this property in both breads. Only one of the two Lactobacillus sanfranciscencis strains was able to improve all functional properties in both GF breads. Back-slopping of the sourdoughs revealed stable properties in case of buckwheat, while maturity of the millet sourdough could not be reached. These observations were supported by the microbial count, metabolite production and carbohydrate consumption. Mature sourdough significantly improved the crumb firmness and porosity of the GF breads. These results highlighted the importance of selecting the appropriate lactic acid bacteria strains, to maximize the quality of GF bread.

In cheese production, knowledge of clostridial spore levels in milk is crucial to avoid economic losses due to late-blowing, a characteristic quality defect observed especially in hard cheeses. However, no international standard method is available to quantify clostridial spores in milk. Since performance data on the currently applied enumeration methods have not been systematically investigated, in this study we assessed four routine media and most probable number (MPN) methods for the enumeration of cheese-damaging clostridia in milk. Among the four evaluated methods, statistically significant differences became evident, while moderate to optimal correlations were observed. Bacterial isolates from positive reactions were identified using molecular methods. All four methods assessed showed a severe lack of selectivity due to the incorporation of non-clostridial spore-formers in the counts produced. The results illustrate major drawbacks of currently used methods and stress the necessity for improving method selectivity, sensitivity and reliability before standardisation can be envisaged.

Lactic acid bacteria (LAB) from spontaneous rye sourdough were isolated, identified, and characterized by their growth, acidification rate, and carbohydrate metabolism. The isolated LAB were used for production of rye sourdough, and the influence of sourdough on mixed rye - wheat bread quality and acrylamide formation was evaluated. In addition, comparative studies by using acidification with L-(+)-lactic acid for mixed rye – wheat bread production were performed. Isolated LAB (P. acidilactici, L. plantarum, L. curvatus) demonstrated versatile carbohydrate metabolism, grown at 30 °C and 37 °C, and acidic tolerance. When the isolated strains were used for rye sourdough production, they showed good growth, acidification rates while excreting amylolytic and proteolytic enzymes. Rye sourdoughs delay bread staling, and there was a significant effect of type of dough acidification (with LAB or L-(+)-lactic acid) and the quantity of acidification agent used (5% or 15%) on most of the analyzed bread quality parameters. L. plantarum sourdough (added in the amount of 5% and 15%) decreased the acrylamide content in bread samples (p

A Lactobacillus plantarum strain was used for the production of rye, wheat, barley, and oat sourdoughs, and the influence of different sourdoughs on mixed rye–wheat bread quality parameters and acrylamide formation was evaluated. L. plantarum LUHS135 demonstrated versatile carbohydrate metabolism, good growth and acidification rates, and the ability to excrete amylolytic and proteolytic enzymes in various cereal sourdoughs. The same starter and different cereal substrates allow to produce sourdoughs showing different characteristics. The type of sourdough and its quantity had significant influence on acrylamide content in bread (P ≤ 0.0001), and using 5% or 10% of wheat sourdough, 5%, 15%, or 20% of barley sourdough, and 5% or 15% of oat sourdough, it is possible to reduce acrylamide content in bread. Thus, manufacturers need to take into account application of apparent technological approaches for acrylamide in bread reducing.

One of the most severe quality defects in hard and semi-hard cheese, the late blowing defect, is caused by endospore-forming bacteria of the genus Clostridium. To minimize financial losses and waste of resources due to cheese spoilage, raw milk with elevated clostridial spore counts should not be used for the production of certain cheese types. In this context, threshold values of clostridial spore concentrations that cause quality defects in cheese are still under debate. To improve our understanding about late blowing defects, further information on the correlation between clostridial spore concentrations in milk and cheese quality is indispensable. Thus, the aim of this study was to monitor the microbiological quality of milk used for Alpine cheese production regarding clostridial endospore levels to facilitate the establishment of threshold spore concentrations that guarantee the absence of quality defects in Austrian cheese. For this purpose, we monitored clostridial endospore levels in vat milk of 4 Alpine dairies throughout the summer grazing period in 2018. Surprisingly, we observed almost complete absence of butyric acid-producing clostridia in milk and no blowing defects in cheese. Hence, critical clostridial spore concentrations could not be verified. Moreover, the observed low spore levels reveal that the prohibition of silage feeding and good farming practices effectively minimize clostridial endospore counts in milk and ensure the manufacture of high-quality cheese even if technological possibilities are limited.

Biofilm characteristics of Microbacterium lacticum D84 (M. lacticum) and Staphylococcus capitis subsp. capitis (S. capitis) on polytetrafluoroethylene and AISI-304 stainless steel at early- (24, 48 h) and late-stage (144, 192 h) biofilm formation were investigated. M. lacticum biofilm structure was more developed compared to S. capitis, representing vastly mature biofilms with a strongly developed amorphous matrix, possibly extracellular polymeric substances (EPSs), at late-stage biofilm formation. S. capitis showed faster growth behavior but still resulted in a relatively flat biofilm structure. Strong correlations were found between several roughness parameters and S. capitis surface coverage (r ≥ 0.98), and between total surface free energy (γs) and S. capitis surface coverage (r = 0.89), while M. lacticum remained mostly unaffected. The pronounced ubiquitous biofilm characteristics make M. lacticum D84 a suitable model for biofilm research. Studying biofilm formation of these bacteria may help one understand bacterial adhesion on interfaces and hence reduce biofilm formation in the food industry.

For decades, microorganisms in beet sugar production have been studied using culture-based methods. However, these methods are not sufficient to describe such a complex bacterial community. In this study, therefore, an amplicon-based sequencing technique (Illumina MiSeq platform) was applied to characterize the bacterial community and its dynamics in the extraction area and juice purification station of an Austrian beet sugar plant. Depending on the process conditions thermophilic bacteria, such as Geobacillus spp., Caenibacillus spp., and Thermus spp., and mesophilic bacteria, such as Leuconostoc spp. and Bacillus spp., were found. Besides these microbiological characteristics, the antimicrobial effect of a rosin acid-based product (Defostab 220) on the bacterial communities was investigated in industrial and laboratory trials. The antimicrobial effect of a given concentration of rosin acid varies from bacteriostatic to bactericidal effects on different occurring groups of bacteria.

Via butyric acid fermentation, clostridia – mainly Clostridium tyrobutyricum – are able to transform lactic acid into butyric acid, acetic acid and gas (H2 and CO2). The presence of clostridial endospores in milk may lead to severe quality defects in semi-hard and hard cheeses. As a consequence of butyric acid fermentation during ripening, cheeses tend to swell and develop undesired slits, irregular eyes and a rancid taste, thus resulting in high economic losses for producers. Several measures regarding stable, milking and feed hygiene have already been partly implemented to minimise the risk of raw milk contamination with clostridial endospores. Contamination, nevertheless, cannot be avoided completely. Moreover, some of the existing procedures to reduce the bacterial and endospore count in milk (e.g. bactofugation, addition of bacteriocins) are not always applicable or even prohibited for the production of certain cheese types. Therefore, cheese producers may benefit from integrating the determination of the initial count of clostridial endospores in milk into their quality control system of primary materials. This review discusses the role of butyric acid clostridia in the cheese processing environment and methods for the detection and enumeration of cheese-damaging clostridia in milk and cheese.

The intestinal microbiota of piglets fed with a Control diet low in dietary fibre and modified wheat bran variants as an additional source of insoluble dietary fibre was characterised. In this context, variances in the microbiota of three different gut segments were assessed. Wheat bran was either included in its native form or modified by fermentation and extrusion before added at 150 g/kg to a basal diet for 48 piglets (12 animals per treatment). Total DNA was extracted from digesta samples from the jejunum, the end of the ileum and the colon ascendens. Samples were prepared accordingly for subsequent sequencing with the Illumina MiSeq. The obtained results revealed distinct location-specific differences in microbial composition. While Firmicutes were most predominant in all three gut segments, Bacteroidetes were additionally found in the colon at high abundance. The parameters of alpha and beta diversity analysis showed significant differences (p

Sourdough processing contributes to better digestible wheat-based bakery products, especially due to the proteolytic activity of lactic acid bacteria (LAB). Therefore, sourdough-related LAB were screened for their capacity to degrade immunogenic proteins like gluten and alpha-amylase-trypsin inhibitors (ATIs). Firstly, the growth of 87 isolates was evaluated on a gluten-based medium. Further, the breakdown capacity of selected isolates was determined for gluten with a focus on gliadins by measuring acidification parameters and MALDI-TOF MS protein profiles. ATI degradation after 72 h of incubation within an ATI-based medium was investigated by means of acidification, HPLC, and competitive ELISA. All isolates exhibited the potential to degrade ATIs to a high degree, whereas the gliadin degradation capacity varied more greatly among tested LAB, with Lacticaseibacillus paracasei Lpa4 exhibiting the strongest alterations of the gliadin pattern, followed by Lactiplantibacillus plantarum Lpl5. ATI degradation capacities ranged from 52.3% to 85.0% by HPLC and 22.2% to 70.2% by ELISA, with Lacticaseibacillus paracasei Lpa4 showing superior breakdown properties. Hence, a selection of specific starter cultures can be used in sourdough processing for wheat-based bakery products with reduced gluten and ATI content and, further, better tolerated products for patients suffering from non-celiac wheat sensitivity (NCWS).

Clostridium tyrobutyricum is well known as one of the main causative agents of severe cheese spoilage. The metabolism of this anaerobic bacterium during ripening leads to textural and sensory defects in cheese and consequential loss of product value. The potential to induce cheese spoilage, however, may vary among different strains of the same species. Therefore, a better understanding of the intra-species diversity of C. tyrobutyricum may be of practical relevance for the dairy industry. In the present study, we compared the ability of three typing techniques to differentiate 95 C. tyrobutyricum strains on the subspecies level: (1) repetitive element palindromic PCR (rep-PCR) fingerprinting combined with conventional agarose gel electrophoresis, (2) hexaplex-PCR followed by an automated capillary electrophoresis and (3) matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) typing. MALDI-TOF MS fingerprinting provided only moderate reproducibility and low discriminatory power. Both PCR-based methods were highly reproducible and discriminative, with hexaplex-PCR fingerprinting being slightly more discriminative than rep-PCR typing. Overall, a high intra-species diversity was observed among the tested strains, indicating that further investigations on the strain level may be of interest.

In sourdough processing, lactic acid bacteria (LAB) and yeasts are responsible for the bread's unique characteristics. To evaluate the microbial ecology of traditional Austrian sourdoughs, 11 wheat and nine rye sourdoughs were examined. The colony counts of the LAB ranged from

Naturally fermented foods are an important part of the typical diet in Cambodia. However, the food safety status of these products has not been widely studied. The aim of this study was, therefore, to provide an overview of the quality of these foods in relation to microbiology and biogenic amines. Additionally, the obtained results were compared to the habits and practices of Cambodians in handling this type of food. A total of 57 fermented foods (42 fishery and 15 vegetable products) were collected from different retail markets in the capital of Cambodia. Pathogenic Salmonella spp., Listeria spp., and Listeria monocytogenes were not detected in 25 g samples. Generally, less than 102 cfu/g of Staphylococcus aureus, Escherichia coli, Pseudomonas spp., Enterobacteriaceae, and molds were present in the fermented foods. Bacillus cereus group members (&lt, 102 to 2.3 &times, 104 cfu/g), lactic acid bacteria (&lt, 102 to 1.1 &times, 107 cfu/g), halophilic and halotolerant bacteria (&lt, 102 to 8.9 &times, 106 cfu/g), sulfite-reducing Clostridium spp. (&lt, 102 to 3.5 &times, 106 cfu/g), and yeasts (&lt, 106 cfu/g) were detected in this study. Still, the presence of pathogenic and spoilage microorganisms in these fermented foods was within the acceptable ranges. Putrescine, cadaverine, tyramine, and histamine were detected in 100%, 89%, 81%, and 75% of the tested products, respectively. The concentrations of histamine (&gt, 500 ppm) and tyramine (&gt, 600 ppm) were higher than the recommended maximum levels in respectively four and one of 57 fermented foods, which represents a potential health risk. The results suggest that the production process, distribution, and domestic handling of fermented foods should be re-evaluated. Further research is needed for the establishment of applicable preservation techniques in Cambodia.

Late-blowing – severe cheese spoilage caused by clostridia – is a long-standing problem of economic importance for dairy processors. However, there is still a paucity of relevant data on clostridia in cheese that would allow some tailored intervention. The main aim of this study was to identify and quantify clostridia in late-blown cheese using a polyphasic methodological approach. Fifty-three cheeses of different origins were analysed using culture-dependent (MPN, enrichment, molecular identification of isolates) and culture-independent methods (real-time PCR, PCR-DGGE and Illumina 16S rDNA sequencing). Using culture-dependent methods, clostridial isolates were collected from several cheese samples, while fewer positive results were obtained with the samples when using the MPN procedure than real-time PCR or PCR-DGGE. In contrast to other studies, an extremely low diversity of Clostridium species was observed. Surprisingly, Illumina 16S rDNA amplicon sequencing revealed relative abundances of clostridia below the detection limit despite apparent cheese spoilage.

Spontaneously fermented wild boar and deer meat sausages produced in Croatia were physicochemically and microbiologically investigated at different time points of their production. Final products resulted with pH and aw values between 5.04–5.53 and 0.83–0.87, respectively. The histamine concentration was below 5.0 mg/kg whereas tyramine content ranged from 47.3 to 219.0 mg/kg. A total of 917 isolated lactic acid bacteria (LAB) were genotyped by rep-PCR and identified by 16S rRNA gene sequencing. Overall, Leuconostoc mesenteroides was identified as the most frequently isolated species (n = 259), followed by Lactobacillus sakei (n = 190) and Enterococcus casseliflavus (n = 106). Cluster analysis revealed a remarkable intraspecies diversity and a strong sausage-specific clustering of the LAB genotypes, indicating meat as a probable source of microbial diversity in sausages. 14.38% of LAB were positive for tyramine encoding gene and no other target biogenic amines genes were detected. Due to the elevated number of presumptive pathogens (E. coli. Enterobacteriaceae, B. cereus group, coliforms) in ready-to-eat sausages, 33.33% of products can be considered as inappropriate for human consumption.

Food packaging materials fundamentally contribute to food quality and safety, as they protect the packaged food against external influences. In this context, the determination of the hygiene status of the packaging material is of great importance. However, European legislation neither sets any microbiological criteria nor provides any approved standard for the microbiological testing of food packaging materials. Nevertheless, reliable routine control is essential for guaranteeing high hygienic quality of packagings. With the aim to achieve a maximum recovery rate at low contamination levels, an improved experimental design was developed for the enumeration of the total colony count, yeasts and molds and Enterobacteriaceae on the surface of roll stock packaging materials. For this purpose, two different types of paper laminates were selected and exemplarily used as objects of investigation. Moreover, the performance of different growth media was compared for each microbiological parameter. This approach was followed by method validation using a selection of quantitative reference materials of representative microorganisms, including resistant forms of microbes such as bacterial endospores and fungal spores.

A pig digestibility trial was conducted to investigate the effects of fermentation or extrusion of wheat bran included in a basal diet on coefficients of total tract apparent digestibility (CTTAD) regarding dry matter (DM), organic matter (OM), crude protein (CP), crude fiber (CF), ether extract (EE), starch, energy (GE), phosphorus (P) and calcium (Ca). In the experiment, 9 growing pigs were allocated to a 3 × 3 Latin square design to measure the CTTAD of the basal diet containing different modified wheat bran variants, and therefore to demonstrate relative differences in the CTTAD among the diets as a result of wheat bran modification. The wheat bran was used in native form (NWB), as fermented bran ensiled with Lactobacillus paracasei and Lactobacillus plantarum (FWB) and as extruded wheat bran (EWB). Wheat bran variants were included at 200 g kg −1 in a phosphorus deficient basal diet. The obtained results show that the CTTAD of DM was increased when feeding the diet with FWB (+2%, P P P P P P P P P P P

The aim of this study is to characterise and examine the biochemical properties of 40 Lactococcus lactis strains isolated from indigenous Montenegrin dairy products in order to explore their potential to be used as starter cultures for producing typical Montenegrin cheese, such as 'bijeli sir', 'masni sir' and 'njeguski sir'. Their safety regarding the production of biogenic amines, the presence of antimicrobial resistance and the antibacterial activity against relevant pathogens and spoilage microorganisms has also been tested. Based on the characterisation, all strains belong to L. lactis ssp. lactis. Out of these 40 strains, 23 displayed rapid acidification ability and proteolysis. However, none of the strains exhibited the ability of lipid degradation. Most of the strains were not associated with any health risk investigated. Summing up, a large percentage (27.5%) of the tested strains showed good properties. These strains should be further examined for their possible application as specific starter cultures in the production of indigenous cheese in Montenegro.

With the increased demand for lightly preserved and/or ready-to-eat (RTE) food products, the prevalence of the foodborne pathogen Listeria monocytogenes has increased, which is a public health concern. The goal for this review is to discuss the incidence, epidemiological importance, and contamination routes of L. monocytogenes in vari- ous aquatic ecosystems, seafood products, and processing environments and to summarize the data obtained since the 1990s. L. monocytogenes primarily enters the food-production chain by cross-contamination in production plants, making this pathogen a major threat to the seafood industry. This pathogen generally contaminates food products at low or moderate levels, but the levels involved in listeriosis outbreaks are significantly higher. The majority of isolates from aquatic products belong to serotype 1/2a, and outbreaks have been linked to highly similar or even indistinguishable strains. Several seafood-processing plants are colonized by specific "in-house" flora containing special DNA subtypes of L. monocytogenes. In such cases, L. monocytogenes populations can persist and/or multiply despite the inherent obstacles to their growth in food preservation and manufacturing operations. Therefore, food-processing facilities must be designed carefully with an emphasis on effective cleaning and disinfecting operations in the production line.

The aim of this study was comparison different methods of species identification of enterococci. One hundred and fifty three suspected colonies were isolated from milk and dairy products (cheeses from cow´s, ewe´s and goat´s milk). On the bases of their growth on BEA agar, microscopic characteristic, results of Gram staining, catalase test and PYRAtest was thirty four isolates assigned to the genus Enterococcus. These isolates were identified by commercial biochemical testEN-COCCUS. 52.9% of them were included in species E. faecalis, 29.4% in E. faecium, 14.7% in E. durans and 2.9% in E. group III. This group includes 3 species: E. durans, E. hirae, E. faecalis asaccharolytic var. Then 16S rRNA sequencing nucleotide of all isolates was realized. Results of sequencing were compared with NCBI database. Only 14.7% of isolates were in 100% accordance. One from them was species E. durans and others were designated as E. faecium. For 20.6% of detected isolates was in accordance with more reference strains. Other isolates were identical with reference strain on 99%. For verification of all results species-specific PCR was used and 52.9% isolates were identified as species E. faecalis, 32.4% as E. faecium and 14.7% as E. durans. Strains belonging to the species E. faecalis were identified the most reliable by all used methods.

Biopreservation is a powerful and natural tool to extend shelf life and to enhance the safety of foods by applying naturally occurring microorganisms and/or their inherent antibacterial compounds of defined quality and at certain quantities. In this context, lactic acid bacteria (LAB) possess a major potential for use in biopreservation because most LAB are generally recognized as safe, and they naturally dominate the microflora of many foods. The antagonistic and inhibitory properties of LAB are due to different factors such as the competition for nutrients and the production of one or more antimicrobially active metabolites such as organic acids (prevailingly lactic and acetic acid), hydrogen peroxide, and antimicrobial peptides (bacteriocins). This review addresses various aspects related to the biological preservation of seafood and seafood products by LAB and their metabolites.

This study aimed to investigate the specific growth stimulation of certain desired intestinal bacteria by a novel galacto-oligosaccharide mixture, which was produced with a &beta, galactosidase from a potential probiotic Lactobacillus isolate that contained mainly oligosaccharides of &beta, 1,3 and &beta, 1,6 glycosidic linkages (termed Lb-GOS) using single-strain fermentations. The composition of this Lb-GOS mixture was 33.5% disaccharides, 60.5% trisaccharides, 4.8% tetrasaccharides, and 1.0% pentasaccharides with a negligible amount of monosaccharides, lactose, and lactobionic acid (0.3%). Eight Lactobacillus spp. strains and three Bifidobacterium spp. strains were used in single-strain fermentations to determine the fermentation activity scores of this Lb-GOS preparation compared to two commercially available prebiotic mixtures, 4&prime, GOS-P and Vivinal GOS (V-GOS). The highest scores were obtained when L. reuteri Lb46 and the two Bifidobacterium strains, B. animalis subsp. lactis Bif1 and Bif3, were grown on these galacto-oligosaccharide mixtures. In addition, the Lb-GOS mixture was found to have higher fermentation activity scores, hence, it stimulated the growth of these probiotic strains more than 4&prime, GOS-P and V-GOS, which may be attributed to the different glycosidic linkage types that are found in the Lb-GOS mixture compared to the other two commercial preparations. These findings suggested that the Lb-GOS mixture that is described in this work should be of interest for the formulations of new carbohydrate-based functional food ingredients.

The antimicrobial spectrum and physico-chemical characteristics of bacteriocin like inhibitory substances produced by lactobacilli isolated from the intestinal flora of Sturgeon fish were determined in order to evaluate their inhibitory potential exerted against 42 food-borne and aquaculture-related bacterial pathogens as well as against food spoilage causing bacteria. In a first series a collection of 84 Lactobacillus strains previously isolated from Beluga (Huso huso) and Persian sturgeon (Acipenser persicus) were screened for their inhibitory activities and potential bacteriocin production against two indicator strains, Listeria monocytogenes ATCC 19115 and Salmonella enterica subsp. enterica serovar Typhimurium ATCC 14028. The isolates Lactobacillus casei AP8 and Lactobacillus plantarum H5 showed the highest activity and therefore were subjected to further examination to clarify the nature of the inhibitory effect. The physico-chemical properties of the harvested antimicrobial compounds were similar to those of bacteriocins of lactobacilli belonging to the group II with respect to molecular weight (5 and 3 kDa respectively), pronounced temperature stability (−20 °C to 120 °C), pH tolerance (3–12), chemical stability (SDS, EDTA, Tween 20, Tween 80) and sensitivity to proteolytic enzymes. Importantly, different food borne pathogens like Escherichia coli, Listeria spp., Salmonella spp., Staphylococcus aureus, Aeromonas hydrophila, Vibrio anguillarum, and Bacillus cereus were inhibited by cell-free supernatants of the strains selected. The broad inhibitory spectrum, the technological properties, especially the stability may lead to the assumption that the bacteriocins like inhibitory AP8 and H5 may be applied as biopreservative agents to control pathogens and spoiling bacteria in different food products. Furthermore their role as bioprotective agents in aquaculture systems is envisaged.

The BacT/ALERT 3D equipment was applied as a fast and sensitive method for the detection of Gram-negative re-contaminants in pasteurised dairy products, particularly drinking milk and cream. The iAST medium was modified and then applied to eleven certified reference strains and nine relevant wild-type isolates originating from a dairy plant. Gram-negative strains could be detected at low levels in pure culture and in combination with high numbers of Gram-positive bacteria as background microflora. Additional experiments with qualified reference material (Escherichia coli, Pseudomonas aeruginosa) showed that low levels of contamination of these bacteria were detectable within 1 and 1.5 days, respectively. The milk matrices exerted no relevant effect on the detectability of the Gram-negative re-contaminants tested. Based on the technique developed, a faster hygiene monitoring and enhanced approval of dairy product lots at the production site is enabled.

In order to obtain good cheese quality, the milk has to possess good physical-chemical properties and should originate from healthy cows. Since milk fat and milk protein are the main constituents of cheese, their proportion in milk is of particular importance for the product yield and quality. This paper describes studies on the protein to fat ratio of milk and the consequent influence on the chemical composition and yield of semi-hard cheese, commercially called "Montenegrin naturally dried cheese". The tests were conducted on six bulk milk samples and six cheese samples. The milk parameters were analysed by the Milkoscan 400 unit whereas chemical analysis of cheese and whey were carried out with the Milkoscan FT 120 device. The average composition of the cheeses was: 29.27% fat, 21.90% protein, 55.27% total dry matter and 0.78% sodium chloride. The mean value for the content of dry matter without fat was 26%, whereas the fat content of the cheese dry matter was 53.18%, the moisture content in cheese 44.73% and moisture content in fat-free matter 63.24%. There was a medium positive correlation between the ratio of fat to protein in milk and fat content in cheese (r = 0.60309), the ratio of fat to protein in milk and dry matter of cheese (r = 0.57103), weak positive correlation between the ratio of fat to protein in milk and cheese protein (r = 0.48067) and medium negative correlation between the ratio of fat to protein in milk and moisture content in cheese (r = -0.57103). Medium negative correlation was found between the ratio of protein to fat in milk and content of cheese fat (r = -0.56416), the ratio of protein to fat in milk and cheese protein content (r = -0.51899), the ratio of protein to fat in milk and dry matter of cheese (r = -0.53118) and medium positive correlation between the ratio of protein to fat in milk and moisture content in cheese (r = -0.53118). Ratio fat to protein in milk and the actual yield of cheese was determined as medium positive (r = 0.66459) and the ratio protein to fat in milk and the actual yield of cheese as medium negative correlation (r = -0.67807). The protein to fat ratio in milk influences the decline of fat, protein, dry matter and yield of cheese and increase moisture content in cheese.