Increase in temperature facilitates Campylobacter jejuni biofilm formation under both aerobic and microaerobic incubation.

The formation of Campylobacter jeuni biofilms on processing surfaces is a significant concern in poultry processing, contributing to food safety risks. This study focused on assessing the biofilm forming capabilities of 12 field isolates of C. jejuni of different aerotolerance categories on stainless steel surfaces, a prevalent material in poultry processing environments. Working cultures of each isolate were prepared to approximately 6 log CFU/mL and incubated on stainless steel coupons under microaerobic or aerobic conditions at room temperature or 42°C for 72 h. Biofilm attached cells were enumerated using direct plating and biofilm density was measured using a crystal violet assay by measuring the optical density (OD600) a. Data analysis was conducted using the PROC GLIMMIX procedure in SAS 9.4 with a significance level of 0.05. The study revealed a notable interaction between aerotolerance categories and temperature (P < 0.039) impacting the number of biofilms attached C. jejuni cells on stainless steel coupons. All isolates had significantly higher counts when incubated at 42°C compared to room temperature, regardless of oxygen level (P < 0.001). Furthermore, stronger biofilm density was observed at 42°C compared to room temperature, regardless of oxygen level. These findings underscore the influence of temperature on the biofilm forming ability of C. jejuni. The ability of these field isolates to form biofilms under various environmental conditions suggests a heightened potential for surface colonization and increased infection risk in poultry processing facilities.

Aerotolerance and Multi-Locus Sequence Typing of Campylobacter jejuni Isolated from Commercial Broiler Processing Plants.

Campylobacter jejuni is one of the leading causes of acute diarrhea in the United States. Despite being a microaerophilic pathogen, C. jejuni continues to endure within the domain of food production, especially in poultry processing. Recent research on aerotolerance indicates that close monitoring of this pathogen is necessary. A total of 40 C. jejuni isolates previously obtained from commercial broiler processing plants were analyzed for aerotolerance and genetic diversity. In addition, the effect of aerotolerance and storage time (days) on the survival of C. jejuni on broiler drumsticks at refrigeration (4 °C) and freezing conditions (-20 °C) was also evaluated. Out of 40 isolates, 25 (62.5%) were aero-sensitive (AS), 10 (25%) were intermediately aerotolerant (IAT), and 5 (12.5%) were hyper aerotolerant (HAT). The isolates belonged to four clonal complexes (CCs) and six sequence types, with the majority of isolates assigned to the CC-353 clonal complex. C. jejuni counts were reduced by 0.40 log CFU/g after 7 days at 4 °C and by 1.50 log CFU/g after 14 days at -20 °C, respectively, irrespective of aerotolerance (p < 0.001). At both refrigeration (p < 0.013) and freezing (p < 0.001), HAT showed greater reductions as compared to AS and IAT. These findings suggest that both refrigeration and freezing reduce C. jejuni counts.

Salmonella Biofilm Formation under Fluidic Shear Stress on Different Surface Materials.

This study characterized biofilm formation of various Salmonella strains on common processing plant surface materials (stainless steel, concrete, rubber, polyethylene) under static and fluidic shear stress conditions. Surface-coupons were immersed in well-plates containing 1 mL of Salmonella (6 log CFU/mL) and incubated aerobically for 48 h at 37 °C in static or shear stress conditions. Biofilm density was determined using crystal violet assay, and biofilm cells were enumerated by plating on tryptic soy agar plates. Biofilms were visualized using scanning electron microscopy. Data were analyzed by SAS 9.4 at a significance level of 0.05. A surface-incubation condition interaction was observed for biofilm density (p < 0.001). On stainless steel, the OD600 was higher under shear stress than static incubation; whereas, on polyethylene, the OD600 was higher under static condition. Enumeration revealed surface-incubation condition (p = 0.024) and surface-strain (p < 0.001) interactions. Among all surface-incubation condition combinations, the biofilm cells were highest on polyethylene under fluidic shear stress (6.4 log/coupon; p < 0.001). Biofilms of S. Kentucky on polyethylene had the highest number of cells (7.80 log/coupon) compared to all other strain-surface combinations (p < 0.001). Electron microscopy revealed morphological and extracellular matrix differences between surfaces. Results indicate that Salmonella biofilm formation is influenced by serotype, surface, and fluidic shear stress.

A Review of Regulatory Standards and Advances in Essential Oils as Antimicrobials in Foods.

As essential oils (EOs) possess GRAS status, there is a strong interest in their application to food preservation. Trends in the food industry suggest consumers are drawn to environmentally friendly alternatives and less synthetic chemical preservatives. Although the use of EOs has increased over the years, adverse effects have limited their use. This review aims to address the regulatory standards for EO usage in food, techniques for delivery of EOs, essential oils commonly used to control pathogens and molds, and advances with new active compounds that overcome sensory effects for meat products, fresh fruits and vegetables, fruit and vegetable juices, seafood, dairy products, and other products. This review will show adverse sensory effects can be overcome in various products by the use of edible coatings containing encapsulated EOs to facilitate the controlled release of EOs. Depending on the method of cooking, the food product has been shown to mask flavors associated with EOs. In addition, using active packaging materials can decrease the diffusion rate of the EOs, thus controlling undesirable flavor characteristics while still preserving or prolonging the shelf life of food. The use of encapsulation in packaging film can control the release of volatile or active ingredients. Further, use of EOs in the vapor phase allows for contact indirectly, and use of nanoemulsion, coating, and film wrap allows for the controlled release of the EOs. Research has also shown that combining EOs can prevent adverse sensory effects. Essential oils continue to serve as a very beneficial way of controlling undesirable microorganisms in food systems.

Global Dietary and Herbal Supplement Use during COVID-19-A Scoping Review.

During the first year of the COVID-19 pandemic, the lack of cure and the intensity of the global spread raised a common awareness of health. The aim of this scoping review is to summarize dietary supplement use globally during first two years of the COVID-19 pandemic. A systematic search was conducted in December 2021 following PRISMA guidelines. PubMed, ERIC, and Scopus databases were searched, and 956 results were screened for eligibility. Fourteen cross-sectional studies from 11 countries and 3 continents were examined. All studies were large population surveys investigating healthy eating and supplement use during COVID-19. Vitamin C, vitamin D, zinc and multivitamins were the most widely reported, as well as natural/herbal products such as ginger and honey. The most common reason cited for supplements use was to strengthen immune system and to prevent infection of COVID-19. These studies reported that populations are relying on healthcare providers, family, friends, and social media to learn about supplement use. Future studies on the treatment of COVID-19 should include more evidence for supplement use.

Roles of Aerotolerance, Biofilm Formation, and Viable but Non-Culturable State in the Survival of Campylobacter jejuni in Poultry Processing Environments.

Campylobacter jejuni is one of the most common causes of foodborne human gastroenteritis in the developed world. This bacterium colonizes in the ceca of chickens, spreads throughout the poultry production chain, and contaminates poultry products. Despite numerous on farm intervention strategies and developments in post-harvest antimicrobial treatments, C. jejuni is frequently detected on broiler meat products. This indicates that C. jejuni is evolving over time to overcome the stresses/interventions that are present throughout poultry production and processing. The development of aerotolerance has been reported to be a major survival strategy used by C. jejuni in high oxygen environments. Recent studies have indicated that C. jejuni can enter a viable but non-culturable (VBNC) state or develop biofilm in response to environmental stressors such as refrigeration and freezing stress and aerobic stress. This review provides an overview of different stressors that C. jejuni are exposed to throughout the poultry production chain and the genotypic and phenotypic survival mechanisms, with special attention to aerotolerance, biofilm formation, and development of the VBNC state.

Microbial Indicators and Possible Focal Points of Contamination during Production and Processing of Catfish.

The catfish industry is important to the United States economy. The present study determined the levels of microbial indicators and the prevalence of Listeria spp. and Listeria monocytogenes at catfish farms and catfish processing plants. Live fish, water, and sediment samples were analyzed in farms. Fish skin, fillets, chiller water, and environmental surfaces were assessed at the processing plants both during operation and after sanitation. Live fish had 2% prevalence of Listeria monocytogenes, while sediment and water were negative for Listeria. Live fish skin counts averaged 4.2, 1.9, and 1.3 log CFU/cm2 aerobic (APC), total coliform (TCC) and generic Escherichia coli counts, respectively. Water and sediment samples averaged 4.8 and 5.8 log CFU/g APC, 1.9 and 2.3 log CFU/g TCC, and 1.0 and 1.6 log CFU/g generic E. coli counts, respectively. During operation, Listeria prevalence was higher in fillets before (57%) and after (97%) chilling than on fish skin (10%). Process chiller water had higher (p ≤ 0.05) APC, TCC, and Listeria prevalence than clean chiller water. After sanitation, most sampling points in which Listeria spp. were present had high levels of APC (>2.4 log CFU/100 cm2). APC combined with Listeria spp. could be a good approach to understand microbial contamination in catfish plants.

Cecal microbiota contribute to the development of woody breast myopathy.

The objective of this study was to characterize the bacterial diversity of cecal microbiota in broilers related to breast phenotype, diet, and genetic strain. Broilers from 2 genetic strains (120 birds/strain) were fed a control diet (15 birds/pen) and an amino acid reduced diet (15 birds/pen, digestible lysine, total sulfur amino acids, and threonine reduced by 20% compared to the control diet). At 8 wk of age, 4 male broilers with normal breast (NB, 1 chick per pen) and 4 male broilers with woody breast (WB, 1 chick per pen) were selected for each treatment (strain × diet). The DNA of cecal samples was extracted and the 16S rRNA genes were sequenced and analyzed. There were no differences (P > 0.05) in the alpha diversity of gut microbiota between 2 phenotypes (NB vs. WB), 2 strains, or 2 diets (control vs. reduced). However, principal coordinate analysis plots (beta diversity) revealed that there were composition differences in samples between the 2 phenotypes (P = 0.001) and the 2 diets (P = 0.024). The most abundant phyla in all samples were Firmicutes, followed by Bacteroidetes and Proteobacteria. There were differences (false discovery rate, FDR < 0.05) in bacterial relative abundance between phenotypes and between diet treatments, but not (FDR > 0.05) between the 2 genetic strains. Selenomonas bovis (12.6%) and Bacteroides plebeius (12.3%) were the top 2 predominant bacteria in the ceca of WB birds; however, the relative abundances of these 2 bacteria were only 5.1% and 1.2% in NB birds, respectively. Function analysis predicted that the metabolic activities differed (q < 0.05) only between phenotypes. The microbiota of WB birds was characterized as reduced glycolysis and urea cycle but increased tricarboxylic acid (TCA) cycles, sugar degradation, and purine and pyrimidine nucleotides biosynthesis. Further studies are needed to investigate if WB incidence could be reduced by regulating gut microbiota and the potential mechanism that leads to decreased WB incidence.

The effect of different ultraviolet-C light doses on microbial reduction and the components of camel milk.

As a result of increasing interest in non-thermal technologies as a possible alternative or complementary to milk pasteurization processing, the objectives of this study were to determine the effects of different ultraviolet-C light doses on the viability of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium and chemical changes to camel milk components. Pasteurized and inoculated camel milk samples were ultraviolet-C treated in a continuous flow system. The viability of E. coli O157:H7 and S. Typhimurium was evaluated with both in vivo imaging system and traditional plate count agar method. Samples subjected to the 4.15, 8.30, and 12.45 mJ/cm2 of ultraviolet-C treatment resulted in 1.9, 3.3, and 3.9-log reductions in E. coli O157:H7 and 0.9, 3, and 3.9-log reductions in S. Typhimurium, respectively. The measurement of secondary lipid peroxidation products (or ThioBarbituric Acid Reactive Substance values) showed no significant (P > 0.05) differences between the raw and ultraviolet-C treated milk samples. Additionally, no changes (P > 0.05) in the protein profiles of αs1-casein, α-lactalbumin, and lactoferrin were observed between both samples. Compared to the untreated raw milk, c9t11 conjugated linoleic acid decreased (P < 0.01) while t10c12 conjugated linoleic acid increased (P < 0.01) in the ultraviolet-C treated milk. Furthermore, three new volatile compounds were identified in the ultraviolet-C treated milk compared to the control. In conclusion, milk treated with the ultraviolet-C light at a dose of 12.45 mJ/cm2 did not meet the Food and Drug Administration (FDA) requirements for the 5-log pathogen reduction. The ultraviolet-C treatment, on the other hand, had minimal effects on camel milk components.

Thyme Oil Enhances the Inactivation of Salmonella enterica on Raw Chicken Breast Meat During Marination in Lemon Juice With Added Yucca schidigera Extract.

Enteric pathogens such as Salmonella enterica can survive in low pH conditions and pose a food safety threat during marinating of raw poultry meat. A study was conducted to investigate the effectiveness of thyme oil for killing S. enterica on raw chicken during marination in lemon juice containing yucca extract. Samples of raw chicken breast were inoculated with a five-serovar mixture of S. enterica (~108 CFU/mL) and immersed for 2, 4, 6, and 8 h in four lemon-based marinades at 22°C: lemon juice alone (L), L with added 0.5% yucca extract (L + Y), L + Y and 0.5% thyme oil (L + Y + 0.5% TO) and L + Y + 1.0% TO. The L and L + Y served as controls. Survivors were determined by surface plating chicken homogenates on xylose-lysine tergitol-4 (XLT4) agar and XLT4 agar overlaid with non-selective agar (TAL) and counting bacterial colonies after 48 h of incubation (35°C). Marinades containing Y and TO significantly reduced initial viable populations of S. enterica compared to control (L and L + Y) solutions (P < 0.05). Based on S. enterica survivors on TAL medium, the L and L + Y reduced initial populations by 1.12 and 1.42 Log CFU/sample, respectively, after 8 h whereas, Log reductions caused by L + Y + 0.5% TO and L + Y + 1.0% TO, respectively, were 2.62 and 3.91 (P < 0.05). Numbers of survivors were higher on TAL compared to XLT4 agar (P < 0.05); however, the extent of sub-lethal injury caused by the marinades was not statistically significant (P > 0.05). The death rate of S. enterica increased significantly (P < 0.05) in the marinades containing TO (0.5 or 1.0%) compared to control (L + Y). Based on these results, thyme oil has good potential to increase the antimicrobial efficacy of lemon juice marinade against Salmonella on raw chicken breast and enhance the microbial safety of this popular poultry product.

Silver-coated magnetic nanocomposites induce growth inhibition and protein changes in foodborne bacteria.

Cytotoxicity concerns of nanoparticles on animal or human bodies have led to the design of iron oxide core nanocomposites, coated with elemental silver to allow their magnetic removal from bio-mixtures. Although the antimicrobial effect of silver is well-described, the effects of nanoparticles derived from silver on microorganisms remain unfolded. Here, we characterized a customized magnetic silver nanocomposite (Ag-MNP) and evaluated its effects on bacterial growth and protein changes. The Ag-MNP displayed both longitudinal and round shapes under High-Resolution Transmission Electron Microscopy imaging, while the Energy Dispersive X-ray Spectroscopy and X-ray diffraction analysis confirmed the presence of Ag, Fe3O4 (Magnetite) and FeO2 (Goethite). Optical density, bioluminescence imaging, and Colony Forming Unit assessments revealed that the presence of Ag-MNP induced strong dose-dependent bacteria (Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium and S. Anatum) growth inhibition. The TEM imaging showed penetration and infiltration of bacteria by Ag-MNP, leading to membrane degeneration and vacuole formation. The presence of Ag-MNP led to fifteen up-regulated and nine down-regulated proteins (P < 0.05) that are involved in cell membrane synthesis, inhibition of protein synthesis, interference with DNA synthesis, and energy metabolism inhibition. This study provides insights to develop alternative antimicrobials to treat foodborne pathogens with antibiotic resistance avoidance.

Real-Time Bioluminescence Analysis of Escherichia coli O157:H7 Survival on Livestock Meats Stored Fresh, Cold, or Frozen.

Foodborne bacteria such as Escherichia coli O157:H7 can cause severe hemorrhagic colitis in humans following consumption of contaminated meat products. Contamination with pathogenic bacteria is frequently found in the food production environment, and adequate household storage conditions of purchased foods are vital for illness avoidance. Real-time monitoring was used to evaluate bacterial growth in ground horse, beef, and pork meats maintained under various storage conditions. Various levels of E. coli O157:H7 carrying the luxCDABE operon, which allows the cells to emit bioluminescence, were used to inoculate meat samples that were then stored at room temperature for 0.5 day, at 4°C (cold) for 7 or 9 days, or -20°C (frozen) for 9 days. Real-time bioluminescence imaging (BLI) of bacterial growth was used to assess bacterial survival or load. Ground horse meat BLI signals and E. coli levels were dose and time dependent, increasing during room temperature and -20°C storage, but stayed at low levels during 4°C storage. No bacteria survived in the lower level inoculum groups (101 and 103 CFU/g). With an inoculum of 107 CFU/g, pork meats had higher BLI signals than did their beef counterparts, displaying decreased BLI signals during 7 days storage at 4°C. Both meat types had higher BLI signals in the fat area, which was confirmed with isolated fat tissues in the beef meat. Beef lean and fat tissues contrasted with both pork fat and lean tissues, which had significantly higher BLI signals and bacterial levels. BLI appears to be a useful research tool for real-time monitoring of bacterial growth and survival in various stored livestock meats. The dependence of E. coli O157:H7 growth on meat substrate (fat or lean) and storage conditions may be used as part of an effective antibacterial approach for the production of safe ground horse, beef, and pork meats.

Antimicrobial Efficacy of Cinnamaldehyde Against Escherichia coli O157:H7 and Salmonella enterica in Carrot Juice and Mixed Berry Juice Held at 4°C and 12°C.

The effectiveness of cinnamaldehyde for inactivating Salmonella enterica and Escherichia coli O157:H7 in carrot juice (CRJ) and mixed berry juice (MBJ) was investigated. Brain heart infusion broth (BHI), CRJ, and MBJ, with concentrations of added cinnamaldehyde ranging from 0.15 to 1.5 µL/mL, 0.25 to 2.0 µL/mL, and 0.25 to 1.5 µL/mL, respectively, were each inoculated with a 5-strain mixture of Salmonella enterica or Escherichia coli O157:H7 to give an initial viable count of 5.07 log10 colony-forming units/mL. Inoculated BHI or juices without cinnamaldehyde served as controls. Growth of the pathogens in BHI (35°C) was monitored by taking absorbance readings (optical density [OD] 600 nm) for 24 h. The inoculated juices were held at 4°C or 12°C for 24 h, and numbers of viable pathogens were determined at 0, 2, 4, 8, and 24 h by plating samples on selective agar followed by incubation (35°C) and counting bacterial colonies at 48 h. The minimum inhibitory concentration of cinnamaldehyde for both pathogens in BHI was 0.25 µL/mL. The pathogens were more sensitive to cinnamaldehyde in MBJ compared with CRJ, irrespective of storage temperature (p < 0.05). At 4°C, cinnamaldehyde (1.5 µL/mL) completely inactivated S. enterica and E. coli in MBJ (negative by enrichment) within 2 h and 8 h, respectively; whereas both pathogens were detected in CRJ (4°C; with 2.0 µL/mL cinnamaldehyde) at 8 and 24 h. At 12°C, S. enterica and E. coli were undetected in MBJ (1.5 µL/mL cinnamaldehyde) within 2 and 4 h, respectively; however, in CRJ (12°C; 2.0 µL/mL cinnamaldehyde), complete inactivation of S. enterica and E. coli occurred within 4 and 24 h, respectively. Cinnamaldehyde is an effective antimicrobial from natural sources that can be used for inactivating bacterial pathogens in fruit and vegetable juices to enhance microbial safety of these nutritious food products.