Carnobacterium Divergens Research Articles

Unusual encounter: Isolation of Carnobacterium divergens in an immunocompetent patient with a meat grinder Injury. Case report

The genus Carnobacterium belongs to lactic acid bacteria, this group are composed by 12 species. These microorganisms are associated with mucosal surface, present in food-related habitats including plant, wine, milk, and meat environments. In humans, evidence of Carnobacterium spp., about the disease spectrum remains unknown. Current work is about a 25-year-old male, was admitted to the emergency room for transmetacarpal traumatic amputation of his left hand caused by a meat grinder. In deep soft tissue samples obtained from surgery, Carnobacterium divergens was isolated. He was treated with piperacillin-tazobactam and was switched to amoxicillin-clavulanate for five more days with clinical success. Written informed consent was obtained from the participating patient for publication of this case report and any accompanying images. Therefore, the pathogenic role of Carnobacterium spp., remains unknown and classically is not considered pathogenic to humans. However, in previous cases where was associated with infection, the described outcomes were good. The misidentification showed that conventional identification methods are not accurate.

High-throughput analysis of microbiomes in a meat processing facility: are food processing facilities an establishment niche for persisting bacterial communities?

BackgroundMicrobial spoilage in meat impedes the development of sustainable food systems. However, our understanding of the origin of spoilage microbes is limited. Here, we describe a detailed longitudinal study that assesses the microbial dynamics in a meat processing facility using high-throughput culture-dependent and culture-independent approaches to reveal the diversity, dispersal, persistence, and biofilm formation of spoilage-associated microbes.ResultsCulture-dependent and culture-independent approaches revealed a large diversity of microbes within the meat facility, including 74 undescribed bacterial taxa and multiple spoilage-associated microbes. Ten out of 10 reconstituted microbial communities formed biofilms, and the biofilm biomass was generally higher at 4 °C than at 25 °C. Isolates obtained at different sampling times or from different sampling sites that differed in fewer than 10 genome-wide single-nucleotide polymorphisms were considered the same (persistent) strains. Strains of Carnobacterium maltaromaticum and Rahnella rivi persisted over a period of 6 months across sampling sites and time, stemming from floor drains in the cooler room. Meat isolates of Carnobacterium divergens, Rahnella inusitata, and Serratia proteamaculans originated from food contact and non-food contact environments of the packaging area.ConclusionsCulture-dependent isolation, complemented by culture-independent analyses, is essential to fully uncover the microbial diversity in food processing facilities. Microbial populations permanently resided within the meat processing facility, serving as a source of transmission of spoilage microbes. The ability of these microbes to coexist and form biofilms facilitates their persistence. Our data together with prior data on persistence of Listeria monocytogenes indicates that microbial persistence in food processing facilities is the rule rather than an exception.-75cynRBG-ecg4-DNt8vX5Video

Potential of Nisin and Newly Discovered Bacteriocins as Preservatives for Pasteurised Milk

Spoilage of pasteurized milk is mainly caused by the presence of organisms that either survive pasteurization (psychrotolerant spore-formers) or re-contaminate milk in the processing environment (post-pasteurisation contaminants). Pasteurization of bovine milk by heat treating at 72°C for 15-30 seconds ensures milk quality without impairing its organoleptic and nutritional status and extends shelf life to 12-14 days at refrigeration temperatures. Nisin A is a class I bacteriocin known to inhibit gram positive bacteria and approved by the European Food and Safety Authority as a food preservative. It is commercially available as Nisaplin®, which contains a concentration of 2.5% w/w of nisin. This study examined the effect of Nisaplin® at different concentrations on spoilage of refrigerated commercial whole pasteurized milk over a period of 59 days. At a high concentration of 4 mgml-1, Nisaplin® reduced the total bacterial count below the limit of detection in the milk, and inhibition was visible for at least 14 days at 4°C. Previously isolated milk spoilage bacteria were identified using 16s rRNA gene sequencing and utilised as target indicators for bacteriocin production. Lactobacillus delbrueckii ssp. bulgaricus LMG6901, Microbacterium lacticum, and Pseudomonas aeruginosa were utilised as indicator strains in a screen of milk spoilage organisms for bacteriocin production. This resulted in identifying the putative bacteriocin producer Carnobacterium divergens, a lactic acid bacterium active against L. bulgaricus. The study concludes that Nisaplin® is effective in the reduction of microbial load and its effectiveness could be increased when combined with other preservative methods thus forming an extra hurdle in the milk.

Solid- and Vapor-Phase Antibacterial Activities and Mechanisms of Essential Oils Against Fish Spoilage Bacteria.

Essential oils (EOs), regarded as secondary metabolites from plants, possess effective antibacterial properties. This study investigates the antibacterial efficacy of seven citrus EOs against six spoilage bacteria: Vibrio parahaemolyticus, V. harveyi, Photobacterium damselae, Shewanella putrefaciens, Carnobacterium divergens, and Lactobacillus pentosus. The antibacterial activity of these EOs was evaluated using solid- and vapor-phase applications. All tested EOs demonstrated effective antibacterial activity at a concentration of 294 μL/L against Gram-negative bacteria. Notably, lemon and orange EOs exhibited dose-dependent inhibition in both solid- and vapor-phase applications, with minimum effective concentrations ranging from 29.4 to 58.8 μL/L. Following treatment with lemon and orange EOs for 6 h at 1/4 minimum inhibitory concentration, leakage of intracellular DNA and proteins was observed, indicating damage to the cell membrane/wall. Proteomic analysis revealed distinct mechanisms: lemon EO impaired bacterial antioxidant defenses, while orange EO disrupted cell division, leading to reduced bacterial viability. These findings provide valuable insights into the potential of different EO application forms in controlling spoilage bacteria.

Comprehensive insights into the organoleptic characteristics attributes of “HuangYuXiang”: Integration of volatilomics, sensomics, macrogenomics, lipidomics, and metabolomics

Comprehensive insights into the organoleptic characteristics attributes of “HuangYuXiang”: Integration of volatilomics, sensomics, macrogenomics, lipidomics, and metabolomics

Antimicrobial Effect of Lippia citriodora Extract in Combination with Gallic Acid or Octyl Gallate on Bacteria from Meat.

Chicken meat and its derivatives are easily alterable. They are a nutritionally healthy food, and their consumption has seen a remarkable increase worldwide in recent years. At the same time, consumer demand for the use of natural products to control microbial growth is increasing. In this context, the antimicrobial capacity of a commercial extract of the lemon verbena (Lippia citriodora) plant, (LCE) was tested in binary combination with gallic acid or octyl gallate against two strains of lactic acid bacteria (LAB) of meat origin: Carnobacterium divergens ATCC 35677 and Leuconostoc carnosum ATCC 49367. First, the antimicrobial potential was evaluated by the checkerboard microdilution method at the optimal growth temperature of each and at 4 °C, pH 5.7 and 6.7, in culture medium. Octyl gallate was the most effective antimicrobial against the two bacteria under all study conditions. At 4 °C, the combination of LCE with octyl gallate had a similar antimicrobial effect on the two LAB, being bactericidal at pH 6.7. In chicken breast, this effective combination was tested in normal or modified atmosphere and refrigerated (4-8 °C) for 9 days. LCE + OG in modified atmosphere reduced the different microbial groups studied, including the lactic acid bacteria as the main microorganisms responsible for the spoilage of fresh meat. Further research could pave the way for the development of novel strategies contributing to the technological stability, security, and functional properties of chicken meat.

Assessment of the Spoilage Microbiota and the Growth Potential of Listeria monocytogenes in Minced Free-Range Chicken Meat Stored at 4 °C in Vacuum: Comparison with the Spoilage Community of Resultant Retail Modified Atmosphere Packaged Products

Although current diet and nutrition trends in developed countries led the poultry industry to shift to alternative breeding/production methods, such as organic and free-range, limited data on the microbiology of alternative compared to conventional poultry meat products exist. Therefore, this study assessed the evolution and composition of the spoilage microbiota and the growth potential of inoculated (3 log cfu/g) Listeria monocytogenes in freshly minced free-range chicken meat stored at 4 °C in vacuum packages (VP; four batches) for 0, 3, 5, 7, and 10 days. Additionally, two VP batches were compared with their resultant retail products stored in modified atmosphere packages (MAP 30:70 CO2/N2) at 4 °C to detect potential differences with the MAP spoilage community described previously. The initial pH of the VP minces was 6.0–6.1, except for one mince, designated VP + AA, which had initial pH 5.8 and was found to contain ‘external’ 1.26% L-lactate and 0.24% acetate associated with a vinegar smell during storage. The rest of the VP batches contained on average 0.75% L-lactate and 0.02% acetate on day 0. After 7 days at 4 °C, L-lactate decreased by at least 3-fold in VP and over 5-fold in VP + AA vs. minor decreases in MAP. Acetate increased 2-fold in all batches. D-lactate (ca. 0.02% on day 0) increased by 4-fold in VP batches only. Lactic acid bacteria (LAB) became the dominant spoilers in all samples. Only VP allowed a delayed 10-fold growth (>5.0 to 6.2 log cfu/g) of pseudomonads from day 7 to day 10 at 4 °C. Compared to VP, VP + AA and MAP retarded growth of LAB, pseudomonads, and enterobacteria by 1–2 log units, at final levels below 6.5, 4.5, and 3.0 log cfu/g, respectively. Enterococci, staphylococci, yeasts, and L. monocytogenes did not grow. Latilactobacillus sakei predominated in all spoiled VP batches (65.8% of 80 meat isolates) followed by Latilactobacillus fuchuensis (9.2%), Leuconostoc carnosum (6.6%), Carnobacterium divergens (6.6%), Latilactobacillus curvatus (5.3%), and Weissella koreensis (2.6%). VP + AA favored Latilactobacillus. Brochothrix thermosphacta was frequent in one VP batch. In conclusion, cold-stored (4 °C), minced, free-range chicken meat spoils more rapidly and offensively under VP than MAP or VP combined with acetate-containing (VP + AA) antimicrobial blends.

Evaluation of probiotic growth stimulation using prebiotic ingredients to optimize compounds for in ovo delivery.

The use of probiotics, prebiotics and synbiotics in poultry diets beneficially stimulates the gut microbiome thus promoting the health and welfare of the animals. In this study, we analyzed 7 poultry probiotics (Lactobacillus plantarum - B1 and B4, Lactobacillus rhamnosus - B3, Bifidobacterium lactis - B2, Carnobacterium divergens - B5, Propionibacterium thoenii - B6, Clostridium butyricum - B7) and 12 prebiotics, differing in chemical composition and source of origin (fungi, algae, animal, etc.). The main goal of our research was to select the most promising candidates to develop synbiotic combinations. We determined the growth kinetics of all probiotics in the presence of prebiotics in a series of in vitro studies to select optimal combinations. Five out of seven investigated probiotics were significantly stimulated by astragalus polysaccharide, and this prebiotic was characterized in our work as the most effective. Moreover, in the case of three probiotics, B2, B3 and B4, significant growth stimulation has been found when beta-glucan, vegetable protein hydrolysate and liquid seaweed extract were supplied. Strain B1 (L. plantarum) was stimulated by 6 out of 12 prebiotics. The growth of B4 (L. plantarum) and B2 (B. lactis) was enhanced by prebiotics after 2 h of incubation. A high growth rate of 3.13% was observed in the case of L. plantarum (B4) and a 3.37% higher rate for B. lactis (B3), compared to the growth of probiotics in the control medium with glucose but no prebiotics. The best candidates for synbiotic combinations based on this in vitro work are the strains belonging to L. plantarum (B4), L. rhamnosus (B3) and B. lactis (B2), consistent with prebiotics such as astragalus polysaccharides and vegetable protein hydrolysate. These combinations will be subject to future in vivo poultry trials involving the in ovo microbiome modulation.

Application of combined essential oils and bacteriocins encapsulated in gelatin for bio‐preservation of meatballs

Abstract In this study, gelatin‐based encapsulation of different bioactive compounds including essential oils (EOs) and bacteriocins, produced by lactic acid bacteria was established to evaluate the microbial, physiochemical, and sensory qualities of meatballs. Determination of minimum inhibitory concentration followed by checkboard method showed citrus extract, Mediterranean formulation,Cinnamonand thyme EOs had inhibitory concentrations between 20 and 5000 ppm and synergistic effect against common contaminant and pathogenic bacteria in meat. The bacteriocins produced byLactobacillus curvatusandPediococcus acidilacticishowed antimicrobial activity between 10,000 and 80,000 ppm againstLeuconostoc mesenteroides,Carnobacterium divergens,Lactobacillus curvatus,Listeria inocua,Listeria monocytogenes, andPseudomonas aeruginosa. Encapsulation of the bioactive compounds in gelatin kept the bioactive content to greater extent. The encapsulated bioactive compounds were effective to inhibit the microbial growth, retard the lipid oxidation and color changes, and preserve the sensorial attributes of meatballs. It can be concluded that gelatin‐based encapsulation of Cinnamon EOs and bacteriocins is effective to extend the shelf‐life of meatballs.

Abolishment of spawner-isolated mortality virus and where the remaining science leads.

In the late 1980s, there was histological and electron microscopy evidence for a parvovirus-like virus in Australian prawns. The data were consistent with infectious hypodermal and haematopoietic necrosis virus (IHHNV). However, these cases did not fit the then current paradigms of the known viruses and sequencing did not find any meaningful sequence homology. The virus was named spawner-isolated mortality virus (SMV; GenBank AF499102.1) in order to allow publication of the information about its occurrence to inform the scientific and aquacultural communities. This virus was present in the early years of mid-crop mortality syndrome (1993-1995). However, as time passed, nucleotide and protein databases have expanded and sequence investigation tools have become more cost effective. The sequence of the entity known as SMV is now shown to be of Carnobacterium divergens (CP016843.1). Therefore, the publications with regard to SMV have been assessed and a recommendation to abolish the name with the still valid science transferred to IHHNV and C. divergens.

Characterization of Bacteriophage cd2, a Siphophage Infecting Carnobacterium divergens and a Representative Species of a New Genus of Phage.

Carnobacterium divergens is frequently isolated from natural environments and is a predominant species found in refrigerated foods, particularly meat, seafood, and dairy. While there is substantial interest in using C. divergens as biopreservatives and/or probiotics, some strains are known to be fish pathogens, and the uncontrolled growth of C. divergens has been associated with food spoilage. Bacteriophages offer a selective approach to identify and control the growth of bacteria; however, to date, few phages targeting C. divergens have been reported. In this study, we characterize bacteriophage cd2, which we recently isolated from minced beef. A detailed host range study reveals that phage cd2 infects certain phylogenetic groups of C. divergens. This phage has a latent period of 60 min and a burst size of ~28 PFU/infected cell. The phage was found to be acid and heat sensitive, with a complete loss of phage activity when stored at pH 2 or heated to 60°C. Electron microscopy shows that phage cd2 is a siphophage, and while it shares the B3 morphotype with a unique cluster of Listeria and Enterococcus phages, a comparison of genomes reveals that phage cd2 comprises a new genus of phage, which we have termed as Carnodivirus. IMPORTANCE Currently, very little is known about phages that infect carnobacteria, an important genus of lactic acid bacteria with both beneficial and detrimental effects in the food and aquaculture industries. This report provides a detailed characterization of phage cd2, a novel siphophage that targets Carnobacterium divergens, and sets the groundwork for understanding the biology of these phages and their potential use in the detection and biocontrol of C. divergens isolates.

Optimization of a natural antimicrobial formulation against potential meat spoilage bacteria and food-borne pathogens: Mixture design methodology and predictive modeling.

Optimization of a natural antimicrobial formulation against potential meat spoilage bacteria and food-borne pathogens: Mixture design methodology and predictive modeling.

Isolation and Characterization of Lactobacillus crispatus, Lactococcus lactis, and Carnobacterium divergens as Potential Probiotic Bacteria from Fermented Black and Green Olives (Olea europaea): An Exploratory Study.

Table olives are becoming well recognized as a source of probiotic bacteria that might be used to create a health-promoting fermented food product by traditional procedures based on the activities of indigenous microbial consortia present in local environments. Methodology. In the present study, the characterization of probiotic bacteria isolated from mince, chunks, and brine of fermented green and black olives (Olea europaea) was done based on morphological, biochemical, and physiological characteristics. Bacterial isolates demonstrated excellent survival abilities at 25, 37, and 45°C and at a variable range of pH. However, the optimum temperature is 37 and the optimum pH is 7 for all three isolates. An antimicrobial susceptibility pattern was found among these isolates through the disc diffusion method. Most of the isolates were susceptible to streptomycin, imipenem, and chloramphenicol, whereas, amoxicillin showed resistance to these isolates, and variable results were recorded for the rest of the antibiotics tested. The growth of the isolates was optimum with the supplementation of 3% NaCl and 0.3% bile salt. The isolated bacteria were able to ferment skimmed milk into yogurt, hence making it capable of producing organic acid. Isolates of Lactobacillus crispatus MB417, Lactococcus lactis MB418 from black olives, and Carnobacterium divergens MB421 from green olives were characterized as potential candidates for use as starter cultures to induce fermentation of other probiotic food products.

Intradiaphragmatic abscesses in a wild boar (Sus scrofa): Inspective implications based on anatomopathological evidences

The intradiaphragmatic localization of an abscess is rarely described in humans and in other animal domestic and wild species, and can be caused by penetrative traumas (i.e., firearm injuries). Here we describe two intradiaphragmatic abscesses in a hunted adult male wild boar (Sus scrofa) pluck, associated with adhesion phenomena with the contiguous anatomical structures (pleural, phrenic, and glissonian serosas) and observed during the post mortem inspection, in accordance with the Reg. EU 627/2019. One of these lesions also presented a phreno-abdominal fistula. We found in cytopathological evaluation of the neoformations’ content the presence of spheroidal bacterial soma, characterized by linearly concatenated “Streptococcus-like” aggregation pattern. Furthermore, microbiological assays revealed a polymicrobial pattern characterized by the presence of telluric microorganisms, some of which have a marked pyogenic action (Streptococcus suis type I, Sphingomonas paucimobilis, Carnobacterium divergens, and Lactobacillus sakei). Our results and collected evidence demonstrate the pathogenetic hypothesis of bacterial contamination secondary to penetrative trauma caused by a not-mortal projectile’s wound, defining the inspective behaviour according to the cogent legislation. These lesions, in the reason of their potential relation to toxemia, bacteremia and septicemia phenomena, represent a sanitary risk that impose, from a normative point of view, the total condemnation of the carcass. These inspective implications, originating from the observation and interdisciplinary description of the anatomopathological and microbiological features of the lesions, are directly projected on the normative scenario, providing useful elements to guarantee the health of the consumer.

Clostridium butyricum Potentially Improves Immunity and Nutrition through Alteration of the Microbiota and Metabolism of Elderly People with Malnutrition in Long-Term Care.

Recent research advances examining the gut microbiome and its association with human health have indicated that microbiota-targeted intervention is a promising means for health modulation. In this study, elderly people in long-term care (aged 83.2 ± 5.3 year) with malnutrition (MNA-SF score ≤ 7) were recruited in a community hospital for a 12-week randomized, single-blind clinical trial with Clostridium butyricum. Compared with the basal fluctuations of the control group, an altered gut microbiome was observed in the intervention group, with increased (p < 0.05) Coprobacillus species, Carnobacterium divergens, and Corynebacterium_massiliense, and the promoted growth of the beneficial organisms Akketmanse muciniphila and Alistipes putredinis. A concentrated profile of 14 increased Kyoto Encyclopedia of Genes and Genomes (KEGG) orthologs (KOs) that were enriched in cofactor/vitamin production and carbohydrate metabolism pathways were discovered; the genes were found to be correlated (p < 0.05) with an elevated abundance of plasma metabolites and short-chain fatty acids (SCFAs), unsaturated medium- to long-chain fatty acids (MFA, LFA), carnitines, and amino acids, thus suggesting a coordinated ameliorated metabolism. Proinflammatory factor interferon-gamma (IFN-γ) levels decreased (p < 0.05) throughout the intervention, while the gut barrier tight junction protein, occludin, rose in abundance (p = 0.059), and the sensitive nutrition biomarker prealbumin improved, in contrast to the opposite changes in control. Based on our results obtained during a relatively short intervention time, C. butyricum might have great potential for improving nutrition and immunity in elderly people in long-term care with malnutrition through the alteration of gut microbiota, increasing the abundance of beneficial bacteria and activating the metabolism in SCFA and cofactor/vitamin production, bile acid metabolism, along with efficient energy generation.

Effects of low-energy electron beam irradiation on the shelf-life and quality of vacuum-packaged beef steaks during chilled storage

Effects of low-energy electron beam irradiation on the shelf-life and quality of vacuum-packaged beef steaks during chilled storage

Impact of Putative Probiotics on Growth, Behavior, and the Gut Microbiome of Farmed Arctic Char (Salvelinus alpinus).

Beneficial bacteria promise to promote the health and productivity of farmed fish species. However, the impact on host physiology is largely strain-dependent, and studies on Arctic char (Salvelinus alpinus), a commercially farmed salmonid species, are lacking. In this study, 10 candidate probiotic strains were subjected to in vitro assays, small-scale growth trials, and behavioral analysis with juvenile Arctic char to examine the impact of probiotic supplementation on fish growth, behavior and the gut microbiome. Most strains showed high tolerance to gastric juice and fish bile acid, as well as high auto-aggregation activity, which are important probiotic characteristics. However, they neither markedly altered the core gut microbiome, which was dominated by three bacterial species, nor detectably colonized the gut environment after the 4-week probiotic treatment. Despite a lack of long-term colonization, the presence of the bacterial strains showed either beneficial or detrimental effects on the host through growth rate enhancement or reduction, as well as changes in fish motility under confinement. This study offers insights into the effect of bacterial strains on a salmonid host and highlights three strains, Carnobacterium divergens V41, Pediococcus acidilactici ASG16, and Lactiplantibacillus plantarum ISCAR-07436, for future research into growth promotion of salmonid fish through probiotic supplementation.

Spoilage potential of bacterial species from chilled vacuum-packed lamb

Spoilage potential of bacterial species from chilled vacuum-packed lamb

Metataxonomic signature of beef burger perishability depends on the meat origin prior grinding

Metataxonomic signature of beef burger perishability depends on the meat origin prior grinding

Microbiota Survey of Sliced Cooked Ham During the Secondary Shelf Life.

Sliced cooked ham packaged in a modified atmosphere is a popular ready-to-eat product, subjected to abundant microbial contamination throughout its shelf life that can lead to deterioration of both sensorial properties and safety. In this study, the microbial load and the chemical–physical features of cooked ham of five producers were monitored for a period of 12 days after the opening of the packages (i.e., the secondary shelf life), during which the products were stored in a domestic refrigerator at 5.2 ± 0.6°C. The sensorial properties presented a perceivable decay after 8 days and became unacceptable after 12 days. High-performance liquid chromatography analysis and solid-phase microextraction coupled with gas chromatography profiling of volatile metabolites indicated that lactic acid, ethanol, acetic acid, acetoin, 3-methyl-1-butanol, and 2-3 butanediol were the main metabolites that characterized the evolution of the analyzed cooked ham. The microbiota was monitored by 16S ribosomal RNA gene profiling and culture-dependent techniques. Already at the opening of packages, all the products presented high microbial load, generally dominated by lactic acid bacteria, with evident differences among the products. The increase of lactic acid bacteria somehow protected samples from abundant contamination by other bacteria, concurring with the evolution of more safe products. This role was exerted by numerous Latilactobacillus, Leuconostoc, and Carnobacterium species, among which the most frequently detected were Latilactobacillus sakei, Latilactobacillus sakei carnosum, Leuconostoc mesenteroides, and Carnobacterium divergens. Some products presented more complex communities that encompassed Proteobacteria such as Moellerella wisconsensis, Proteus hauseri, Brochothrix thermosphacta, and less frequently Pseudomonas, Erwinia, and Massilia. Opportunistic pathogenic bacteria such as Escherichia coli and Vibrio sp. were found in small quantities. The yeasts Kazachstania servazzii and Debaryomyces hansenii occurred already at 0 days, whereas various species of Candida (Candida zeylanoides, Candida sake, Candida norvegica, and Candida glaebosa) were abundant only after 12 days. These results indicated that the microbiological contaminants overgrowing during the secondary shelf life did not derive from environmental cross-contamination at the opening of the tray but were already present when the packages were opened, highlighting the phases of production up to the packaging as those crucial in managing the safety risk associated to this product.