Category Archives: Lactobacillus

Research – Microbial Properties of Raw Milk throughout the Year and Their Relationships to Quality Parameters


Raw milk microbiota is complex and influenced by many factors that facilitate the introduction of undesirable microorganisms. Milk microbiota is closely related to the safety and quality of dairy products, and it is therefore critical to characterize the variation in the microbial composition of raw milk. In this cross-sectional study, the variation in raw milk microbiota throughout the year (n = 142) from three farms in China was analyzed using 16S rRNA amplicon sequencing, including α and β diversity, microbial composition, and the relationship between microbiota and milk quality parameters. This aimed to characterize the contamination risk of raw milk throughout the year and the changes in quality parameters caused by contamination. Collection month had a significant effect on microbial composition; microbial diversity was higher in raw milk collected in May and June, while milk collected in October and December had the lowest microbial diversity. Microbiota composition differed significantly between milk collected in January–June, July–August, and September–December (p < 0.05). Bacterial communities represented in raw milk at the phylum level mainly included Proteobacteria, Firmicutes and Bacteroidota; PseudomonasAcinetobacterStreptococcus and Lactobacillus were the most common genera. Redundancy analysis (RDA) found strong correlations between microbial distribution and titratable acidity (TA), fat, and protein. Many genera were significantly correlated with TA, for example Acinetobacter (R = 0.426), Enhydrobacter (R = 0.309), Chryseobacterium (R = 0.352), Lactobacillus (R = −0.326), norank_o__DTU014 (R = −0.697), norank_f__SC-I-84 (R = −0.678), and Subgroup_10 (R = −0.721). Additionally, norank_f__ Muribaculaceae was moderately negatively correlated with fat (R = −0.476) and protein (R = −0.513). These findings provide new information on the ecology of raw milk microbiota at the farm level and contribute to the understanding of the variation in raw milk microbiota in China. View Full-Text

Research – Evaluation of the safety and efficacy of lactic acid to reduce microbiological surface contamination on carcases from kangaroos, wild pigs, goats and sheep


Studies evaluating the safety and efficacy of lactic acid to reduce microbiological surface contamination from carcases of wild game (i.e. kangaroos and wild pigs) and small stock (i.e. goats and sheep) before chilling at the slaughterhouse were assessed. Wild pig and kangaroo hide‐on carcases may have been chilled before they arrive at the slaughterhouse and are treated after removal of the hides. Lactic acid solutions (2–5%) are applied to the carcases at temperatures of up to 55°C by spraying or misting. The treatment lasts 6–7 s per carcass side. The Panel concluded that: [1] the treatment is of no safety concern, provided that the lactic acid complies with the European Union specifications for food additives; [2] based on the available evidence, it was not possible to conclude on the efficacy of spraying or misting lactic acid on kangaroo, wild pig, goats and sheep carcases; [3] treatment of the above‐mentioned carcases with lactic acid may induce reduced susceptibility to the same substance, but this can be minimised; there is currently no evidence that prior exposure of food‐borne pathogens to lactic acid leads to the occurrence of resistance levels that compromise antimicrobial therapy; and [4] the release of lactic acid is not of concern for the environment, assuming that wastewaters released by the slaughterhouses are treated on‐site, if necessary, to counter the potentially low pH caused by lactic acid, in compliance with local rules.

Research – Role of Lactic Acid Bacteria in Food Preservation and Safety


Fermentation of various food stuffs by lactic acid bacteria is one of the oldest forms of food biopreservation. Bacterial antagonism has been recognized for over a century, but in recent years, this phenomenon has received more scientific attention, particularly in the use of various strains of lactic acid bacteria (LAB). Certain strains of LAB demonstrated antimicrobial activity against foodborne pathogens, including bacteria, yeast and filamentous fungi. Furthermore, in recent years, many authors proved that lactic acid bacteria have the ability to neutralize mycotoxin produced by the last group. Antimicrobial activity of lactic acid bacteria is mainly based on the production of metabolites such as lactic acid, organic acids, hydroperoxide and bacteriocins. In addition, some research suggests other mechanisms of antimicrobial activity of LAB against pathogens as well as their toxic metabolites. These properties are very important because of the future possibility to exchange chemical and physical methods of preservation with a biological method based on the lactic acid bacteria and their metabolites. Biopreservation is defined as the extension of shelf life and the increase in food safety by use of controlled microorganisms or their metabolites. This biological method may determine the alternative for the usage of chemical preservatives. In this study, the possibilities of the use of lactic acid bacteria against foodborne pathogens is provided. Our aim is to yield knowledge about lactic acid fermentation and the activity of lactic acid bacteria against pathogenic microorganisms. In addition, we would like to introduce actual information about health aspects associated with the consumption of fermented products, including probiotics.

Research – Microbial Diversity and Safety in Fermented Beverages


For thousands of years, humans have exploited the natural process of fermentation of various foods to preserve them, and to enjoy the changes in the sensory characteristics that could be produced. Recently, the world of fermented beverages has gone through a rapid transformation linked to deep changes in consumer preferences, consumption habits, climate, new regulations and entry of emerging countries, accompanied by safety concerns and an important reduction in economic resources available to people. As with all food handling and preparation, we need to be sure the fermented food produced is safe. Fermentation is a complex biological process where microbial diversity takes place and the environment created inside of the fermented food provides the conditions to reduce the risk of pathogenic bacteria growth, thus providing safe food. In addition, food manufacturers fermenting food carefully control their processing and must comply with the National Food Standards Codes. Although these products have a generally good food safety record, sometimes inadequate manufacturing practices or the presence of acidophilic pathogens could compromise food safety. In fact, fermented beverages may adversely become contaminated with pathogens or microbial toxins and thereby transform into vehicles that can transmit diseases to the consumers. Moreover, many microorganisms can deteriorate the physical-chemical and sensory properties as well as the flavor of the final products. In this editorial, we present an overview of a review and six original research papers published in the Special Issue “Fermentation Process and Microbial Safety of Beverages” of the Beverages journal.

Research – Enhanced antimicrobial activity against Alicyclobacillus acidoterrestris in apple juice by genome shuffling of Lactobacillus acidophilus NX2-6

Wiley Online

Food safety and quality are seriously compromised by the growth of spoilage bacteria in food. Various lactic acid bacteria (LAB) may inhibit the growth of spoilage bacteria in food and animal feed. In our study, the antibacterial activity of Lactobacillus acidophilus NX2-6 was improved by genome shuffling. The starting populations were obtained via 60Coγ-ray, diethyl sulfate mutagenesis, and UV mutagenesis of protoplasts. The optimal conditions for protoplast formation and regeneration were found to be 30 mg/ml of lysozyme at 37°C for 40 min. The protoplasts were inactivated by UV irradiation for 110 s or by heating at 58°C for 30 min. After two rounds of genome shuffling, the inhibitory activity of strain F50 was improved by 5.6-fold compared to the original strain. The shuffled strain has broad-spectrum antibacterial activity and is a promising candidate for bio-preservative. The antibacterial substances produced by L. acidophilus NX2-6 can be applied to the preservation of apple juice. The results showed that the growth of Alicyclobacillus acidoterrestris in apple juice was significantly inhibited by the addition of 0.1% culture supernatant with acidocin NX2-6 at 28°C and 0.2% culture supernatant with acidocin NX2-6 at 45°C for 30 days. The sugar−acid ratio, pH value, clarity, and fruit flavor of apple juice were well maintained during storage. This study showed that genome shuffling was an effective strategy to improve the bacterial inhibitory activity of LAB and that the shuffled LAB and their metabolites are expected to be used as bio-based food protectors for food safety.

Research – Antagonistic Effects of Conjugated Linoleic Acids of Lactobacillus casei Against Foodborne Enterohemorrhagic Escherichia coli 

Journal of Food Protection

Probiotics in fermented foods or commercially available supplements benefit the host by providing metabolites/peptides. The production of these metabolites varies with available substrates/prebiotic present in the system and their concentration. In this study, 0.5% peanut flour (PF) was used to stimulate the growth and production of metabolites of wild-type Lactobacillus casei (LC wt ) and compare with an engineered L. casei (LC CLA ) capable of converting a higher amount of conjugated linoleic acid (CLA). The total extracellular metabolites present in the cell-free cultural supernatant (CFCS) of LC wt (without peanut), LC wt+PF (with peanut), and LC CLA were collected after 24 h and 48 h of incubation, and their antagonistic activities against enterohemorrhagic Escherichia coli (EHEC EDL933) growth and pathogenesis were evaluated. All the collected metabolites exhibited varying efficiency in restraining EDL933 growth while supplementing low concentration of CLA to the 48-h CFCS from LC wt showed augmented antagonism toward EDL933. A downregulation of key virulence genes was observed from metabolites collected at 48-h time point. These observations indicate that the presence of metabolites in CFCSs including CLA, produced by Lactobacillus , which was further identified by gas chromatography-mass spectrometry; plays a critical role. This study demonstrates the potential applicability of Lactobacillus -originated CLA in the prevention of EDL933 mediated illnesses.

Research – Probiotic Lactobacilli Do Not Protect Chickens against Salmonella Enteritidis Infection by Competitive Exclusion in the Intestinal Tract but in Feed, Outside the Chicken Host


Lactobacilli are commonly used as probiotics in poultry to improve production parameters and to increase chicken resistance to enteric infections. However, lactobacilli do not efficiently colonise the chicken intestinal tract, and also, their anti-infection effect in vivo is sometimes questionable. In this study, we therefore evaluated the potential of a mixture of four Lactobacillus species (L. salivariusL. reuteriL. ingluviei and L. alvi) for the protection of chickens against Salmonella Enteritidis infection. Whenever the chickens were inoculated by lactobacilli and S. Enteritidis separately, there was no protective effect of lactobacilli. This means that when lactobacilli and S. Enteritidis are exposed to each other as late as in the crop of chickens, lactobacilli did not influence chicken resistance to S. Enteritidis at all. The only positive effect was recorded when the mixture of lactobacilli and S. Enteritidis was used for the inoculation of feed and the feed was anaerobically fermented for 1 to 5 days. In this case, chickens fed such a diet remained S. Enteritidis negative. In vitro experiments showed that the protective effect was caused by acidification of feed down to pH 4.6 due to lactobacilli fermentation and was associated with S. Enteritidis inactivation. The probiotic effect of lactobacilli was thus expressed in the feed, outside the chicken host. View Full-Text

Research – Lactic Acid Bacteria as Antimicrobial Agents: Food Safety and Microbial Food Spoilage Prevention


In the wake of continual foodborne disease outbreaks in recent years, it is critical to focus on strategies that protect public health and reduce the incidence of foodborne pathogens and spoilage microorganisms. Currently, there are limitations associated with conventional microbial control methods, such as the use of chemical preservatives and heat treatments. For example, such conventional treatments adversely impact the sensorial properties of food, resulting in undesirable organoleptic characteristics. Moreover, the growing consumer advocacy for safe and healthy food products, and the resultant paradigm shift toward clean labels, have caused an increased interest in natural and effective antimicrobial alternatives. For instance, natural antimicrobial elements synthesized by lactic acid bacteria (LAB) are generally inhibitory to pathogens and significantly impede the action of food spoilage organisms. Bacteriocins and other LAB metabolites have been commercially exploited for their antimicrobial properties and used in many applications in the dairy industry to prevent the growth of undesirable microorganisms. In this review, we summarized the natural antimicrobial compounds produced by LAB, with a specific focus on the mechanisms of action and applications for microbial food spoilage prevention and disease control. In addition, we provide support in the review for our recommendation for the application of LAB as a potential alternative antimicrobial strategy for addressing the challenges posed by antibiotic resistance among pathogens. View Full-Text

Research – Characterization of bacteria and antibiotic resistance in commercially-produced cheeses sold in China

Journal of Food Protection

The consumption of cheese in China is increasing rapidly. Little is known about the microbiota, the presence of antibiotic-resistant bacteria, or the distribution of antibiotic resistance genes (ARGs) in commercially-produced cheeses sold in China. These are important criteria for evaluating quality and safety. Thus, this study assessed the metagenomics of fifteen types of cheese using 16S rRNA gene sequencing. Fourteen bacterial genera were detected. Lactococcus , Lactobacillus , and Streptococcus were dominant based on numbers of sequence reads. Multidrug-resistant lactic acid bacteria were isolated from most of the types of cheese. The isolates showed 100% and 91.7% resistance to streptomycin and sulfamethoxazole, respectively, and genes involved in acquired resistance to streptomycin ( strB) and sulfonamides ( sul2) were detected with high frequency. To analyze the distribution of ARGs in the cheeses in overall, 309 ARGs from eight categories of ARG and nine transposase genes were profiled. A total of 169 ARGs were detected in the 15 cheeses; their occurrence and abundance varied significantly between cheeses. Our study demonstrates that there is various diversity of the bacteria and ARGs in cheeses sold in China. The risks associated with multidrug resistance of dominant lactic acid bacteria are of great concern.

Research – Microbial Growth Study on Pork Loins as Influenced by the Application of Different Antimicrobials


The use of antimicrobials in the pork industry is critical in order to ensure food safety and, at the same time, extend shelf life. The objective of the study was to determine the impact of antimicrobials on indicator bacteria on pork loins under long, dark, refrigerated storage conditions. Fresh boneless pork loins (n = 36) were split in five sections and treated with antimicrobials: Water (WAT), Bovibrom 225 ppm (BB225), Bovibrom 500 ppm (BB500), Fit Fresh 3 ppm (FF3), or Washing Solution 750 ppm (WS750). Sections were stored for 1, 14, 28, and 42 days at 2–4 °C. Mesophilic and psychrotrophic aerobic bacteria (APC-M, APC-P), lactic acid bacteria (LAB-M), coliforms, and Escherichia coli were enumerated before intervention, after intervention, and at each storage time. All bacterial enumeration data were converted into log10 for statistical analysis, and the Kruskal–Wallis test was used to find statistical differences (p < 0.05). Initial counts did not differ between treatments, while, after treatment interventions, treatment WS750 did not effectively reduce counts for APC-M, APC-P, and coliforms (p < 0.01). BB500, FF3, and WS750 performed better at inhibiting the growth of indicator bacteria when compared with water until 14 days of dark storage. View Full-Text