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In modern broilers, the period of embryonic development constitutes a greater proportion of a broiler's productive life. Hence, optimum embryonic development can exert a significant influence not only on chick hatchability and hatchling quality but also on overall broiler growth and performance. Further healthy and active hatchlings are correlated with improved posthatch performance. In this regard, probiotics are good candidates to mediate early-life programming. Therefore, we evaluated the effect of In ovo probiotic spray application on broiler hatchability and hatchling quality. The experiment was set out as a completely randomized study with 2 independent trials. In each trial, 540 eggs (Ross 308) were either sprayed with phosphate buffered saline (PBS; control) or probiotics [∼9 log CFU/egg of Lactobacillus rhamnosus NRRL B-442(LR) or Lactobacillus paracasei DUP 13076 (LP)] during incubation. On day 18, eggs were transferred to the hatcher and set up for hatching. Starting on day 19, eggs were observed for hatching to determine the spread of hatch and hatchability. Hatched chicks were then assessed for quality using the Tona and Pasgar score and morphometric measurements including hatchling weight, yolk-free-body-mass and hatchling length were measured. Further, chicks were reared in floor pens for 3 wk to assess posthatch growth. Overall, In ovo probiotic supplementation improved hatchability and hatchling quality. Specifically, the spray application of LP improved hatchability by ∼ 5% without affecting the spread of hatch. Further, both LR and LP significantly improved Pasgar and Tona score, indicating an improvement in hatchling quality. Also, LP and LR significantly improved hatchling weight, yolk-free-body-mass, and posthatch growth in chicks. LR significantly improved hatchling weight and hatchling length (P < 0.05). Moreover, this increase in posthatch growth was positively correlated with hatchling weight in the probiotic groups. Overall, our study demonstrates that In ovo probiotic application exerts a positive effect on hatchability, hatchling quality, and subsequent posthatch growth., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Controlled environment agriculture (CEA) is a rapidly growing sector that presents unique challenges and opportunities in ensuring food safety. This manuscript highlights critical gaps and needs to promote food safety in CEA systems as identified by stakeholders (n=47) at the Strategizing to Advance Future Extension andResearch (S.A.F.E.R.) CEA conference held in April 2023 at The Ohio State University's Ohio CEA Research Center. Feedback collected at the conference was analyzed using an emergent thematic analysis approach to determine key areas of focus. Research-based guidance is specific to the type of commodity, production system type, and size. Themes include the need for improved supply chain control, cleaning, and sanitization practices, pathogen preventive controls and mitigation methods and training and education. Discussions surrounding supply chain control underscored the significance of the need for approaches to mitigate foodborne pathogen contamination. Effective cleaning and sanitization practices are vital to maintaining a safe production environment, with considerations such as establishing standard operating procedures, accounting for hygienic equipment design, and managing the microbial communities within the system. Data analysis further highlights the need for risk assessments, validated pathogen detection methods, and evidence-based guidance in microbial reduction. In addition, training and education were identified as crucial in promoting a culture of food safety within CEA. The development of partnerships between industry, regulatory, and research institutions are needed to advance data-driven guidance and practices across the diverse range of CEA operations and deemed essential for addressing challenges and advancing food safety practices in CEA. Considering these factors, the CEA industry can enhance food safety practices, foster consumer trust, and support its long-term sustainability., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Probiotics are widely used as feed supplements in the poultry industry to promote growth and performance in chickens. Specifically, this supplementation starts around the time of lay and continues through the production cycle in laying hens. However, the embryonic period is critical to the growth and development of metabolically active organs thereby influencing subsequent health and productivity in adult birds. Therefore, the present study investigated the potential use of probiotics to promote embryonic growth in layers. Further, a pilot grow-out study was conducted to evaluate the effect of in ovo and in-feed probiotic application on pullet growth. For the study, fertile White Leghorn eggs were sprayed with phosphate buffered saline (control, CON) or probiotic cocktail (in ovo only, IO; Lactobacillus paracasei DUP 13076 and L. rhamnosus NRRL B 442) prior to and during incubation. The embryos were sacrificed on d 7, 10, 14, and 18 of incubation for embryo morphometry. On d 18, remaining eggs were set in the hatcher to assess hatchability and hatchling morphometry. For the pullet trial, hatchlings were raised on feed with or without probiotics until wk 5. Pullets were sacrificed weekly, and morphometric parameters were recorded. Results of our study demonstrate that in ovo probiotic application significantly improved relative embryo weight, crown-rump length, hatchability, and hatchling weight when compared to the control (P < 0.05). Further, this enhanced embryonic development was associated with a concomitant increase in posthatch growth. Specifically, pullets raised from probiotic-sprayed eggs had significantly improved crown-rump length, tibial length, tibial bone weight, and body weight when compared to the control (P < 0.05). Moreover, among the different treatment schemes employed in this study [CON (no probiotics), in-feed only (IF), IO only, and in ovo and in-feed probiotic supplementation (IOIF)], sustained probiotic supplementation (IOIF) was found to be the most effective in promoting growth. Therefore, in ovo and in-feed probiotic supplementation could be employed to promote embryo and pullet growth to support subsequent performance in layers., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

In chickens, muscle development during embryonic growth is predominantly by myofiber hyperplasia. Following hatch, muscle growth primarily occurs via hypertrophy of the existing myofibers. Since myofiber number is set at hatch, production of more muscle fibers during embryonic growth would provide a greater myofiber number at hatch and potential for posthatch muscle growth by hypertrophy. Therefore, to improve performance in broilers, this study investigated the effect of in ovo spray application of probiotics on overall morphometry and muscle development in broiler embryos. For the study, fertile Ross 308 eggs were sprayed with different probiotics; Lactobacillus paracasei DUP 13076 (LP) and L. rhamnosus NRRL B 442 (LR) prior to and during incubation. The embryos were sacrificed on d 7, 10, 14, and 18 for embryo morphometry and pectoralis major muscle (PMM) sampling. Muscle sections were stained and imaged to quantify muscle fiber density (MFD), myofiber cross-sectional area (CSA), and nuclei density. Additionally, gene expression assays were performed to elucidate the effect of probiotics on myogenic genes. In ovo probiotic supplementation was found to significantly improve embryo weight, breast weight, and leg weight (P < 0.05). Further, histological analysis of PMM revealed a significant increase in MFD and nuclei number in the probiotic-treated embryos when compared to the control (P < 0.05). In 18-day-old broiler embryos, myofibers in the treatment group had a significantly smaller CSA (LP: 95.27 ± 3.28 μm 2 , LR: 178.84 ± 15.1 μm 2 ) when compared to the control (211.41 ± 15.67 μm 2 ). This decrease in CSA was found to be associated with a concomitant increase in MFD (fibers/mm 2 ) in the LP (13,647 ± 482.15) and LR (13,957 ± 463.13) group when compared to the control (7,680 ± 406.78). Additionally, this increase in myofibrillar hyperplasia in the treatment groups was associated with upregulation in the expression of key genes regulating muscle growth including MYF5, MYOD, MYOG, and IGF-1. In summary, in ovo spray application of probiotics promoted overall embryo growth and muscle development in broilers., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Recent recalls of stone fruit due to potential Listeria contamination and associated foodborne outbreaks highlight the risk for pathogen transmission through stone-fruit consumption. Particularly, surface contamination of fruits increases the risk for cross-contamination of produce during processing and storage. This highlights the need for quality control in stone fruits intended for consumption. To develop effective food safety practices, it is essential to determine the critical factors during stone-fruit processing that influence Listeria survival. Therefore, this study evaluated the ability of Listeria to survive on peaches and nectarines under simulated stone-fruit loading and staging, waxing and fungicide application and storage conditions. The results of our study indicate that current stone-fruit handling conditions do not favor Listeria growth. However, once fruit is contaminated, Listeria can survive on the fruit surface in significant numbers under current processing conditions. Therefore, there is a need to develop and implement preventive controls at the stone-fruit packinghouse to prevent Listeria contamination and deter pathogen persistence.

Consumption of raw mangoes has led to multiple Salmonella- associated foodborne outbreaks in the United States. Although several studies have investigated the epiphytic fitness of Salmonella on fresh produce, there is sparse information available on the survival of Salmonella on mangoes under commercial handling and storage conditions. Hence, the objective of the study was to evaluate the survival of Salmonella on mangoes under ambient conditions simulating the mango packing house and importer facility. Further, the ability of the pathogen to adhere and attach on to the mango fructoplane was also investigated. For the attachment assays, mango skin sections were inoculated with fifty microliters of S. Newport suspension (6.5 log CFU/skin section) and minimum time required for adhesion and attachment were recorded. With the survival assays, unwaxed mangoes were spot inoculated with the Salmonella cocktail to establish approximately 4 and 6.5 log CFU/mango. The fruits were then subjected to different storage regimens simulating fruit unloading, waxing, and storage at the packing house and ripening and storage at the importer facility. Results of our study reveal that Salmonella was able to adhere on to the fructoplane immediately after contact. Further, formation of attachment structures was seen as early as 2 min following inoculation. With the survival assays, irrespective of the inoculum levels, no significant increase or decrease in pathogen population was observed when fruit were stored either at ambient (29-32°C and RH 85-95%, for 48 h), ripening (20-22°C and RH 90-95% for 9 days) or refrigerated storage (10-15°C and 85-95% for 24-48 h) conditions. Therefore, once contaminated, mangoes could serve as potential vehicles in the transmission of Salmonella along the post-harvest environment. Hence development and adoption of effective food safety measures are warranted to promote the microbiological safety of mangoes.

Urinary tract infections (UTI) are the most common hospital-acquired infections in humans and are caused primarily by uropathogenic Escherichia coli (UPEC). Indwelling urinary catheters become encrusted with UPEC biofilms that are resistant to common antibiotics, resulting in chronic infections. Therefore, it is important to control UPEC biofilms on catheters to reduce the risk for UTIs. This study investigated the efficacy of selenium for inhibiting and inactivating UPEC biofilms on urinary catheters. Urinary catheters were inoculated with UPEC and treated with 0 and 35 mM selenium at 37 °C for 5 days for the biofilm inhibition assay. In addition, catheters with preformed UPEC biofilms were treated with 0, 45, 60, and 85 mM selenium and incubated at 37 °C. Biofilm-associated UPEC counts on catheters were enumerated on days 0, 1, 3, and 5 of incubation. Additionally, the effect of selenium on exopolysacchride (EPS) production and expression of UPEC biofilm-associated genes was evaluated. Selenium at 35 mM concentration was effective in preventing UPEC biofilm formation on catheters compared to controls ( p < 0.05). Further, this inhibitory effect was associated with a reduction in EPS production and UPEC gene expression. Moreover, at higher concentrations, selenium was effective in inactivating preformed UPEC biofilms on catheters as early as day 3 of incubation. Results suggest that selenium could be potentially used in the control of UPEC biofilms on urinary catheters.

The multistate Escherichia coli ( E. coli ) O157:H7 outbreak associated with in-shell hazelnuts highlights the pathogen's ability to involve non-traditional vehicles in foodborne infections. Furthermore, it underscores significant gaps in our knowledge of pathogen survivability and persistence on nuts. Therefore, this study investigated the ability of E. coli O157:H7 to attach and survive on in-shell hazelnuts. In-shell hazelnuts were inoculated with a four-strain mixture of E. coli O157:H7 at 7.6 log colony forming units (CFU)/nut by wet or dry inoculation, stored at ambient conditions (24 ± 1 °C; 40% ± 3% relative humidity (RH) and sampled for twelve months. For the attachment assay, in-shell hazelnuts were inoculated and the adherent population was enumerated at 30 s -1 h following inoculation. Irrespective of the inoculation method, ~5 log CFU of adherent E. coli O157:H7 was recovered from the hazelnuts as early as 30 s after inoculation. Conversely, pathogen survival was significantly reduced under dry inoculation with samples being enrichment negative after five months of storage ( p < 0.05). On the other hand, wet inoculation led to a significantly longer persistence of the pathogen with ~3 log CFU being recovered from the in-shell nuts at 12 months of storage ( p < 0.05). These results indicate that E. coli O157:H7 can survive in significant numbers on in-shell hazelnuts when stored under ambient conditions.

Lactic acid bacteria are known to exhibit probiotic properties through various mechanisms, including competitive exclusion, pathogen inhibition, production of antimicrobial substances, and maintenance of eubiosis. Here, we present the draft genome sequence of a novel probiotic strain, Lactobacillus rhamnosus strain NRRL B-442, which exhibits potent antivirulence activity against Salmonella enterica ., (Copyright © 2018 Muyyarikkandy et al.)

Lactobacillus paracasei DUP 13076 demonstrates antagonistic effects against the foodborne pathogens Salmonella enterica serovars Enteritidis, Typhimurium, and Heidelberg in coculture and in vitro experiments. Here, we report the draft genome sequence of Lactobacillus paracasei DUP 13076, which has a circular chromosome of 3,048,314 bp and a G+C content of 46.3%., (Copyright © 2018 Muyyarikkandy et al.)

Liquid phase exfoliation of graphite in six different animal sera and evaluation of its toxicity are reported here. Previously, we reported the exfoliation of graphene using proteins, and here we extend this approach to complex animal fluids. A kitchen blender with a high-turbulence flow gave high quality and maximum exfoliation efficiency in all sera tested, when compared to the values found with shear and ultrasonication methods. Raman spectra and electron microscopy confirmed the formation of three- or four-layer, submicrometer size graphene, independent of the serum used. Graphene prepared in serum was directly transferred to cell culture media without post-treatments. Contrary to many reports, a nanotoxicity study of this graphene fully dispersed to human embryonic kidney cells, human lung cancer cells, and nematodes (Caenorhabditis elegans) showed no acute toxicity for up to 7 days at various doses (50-500 μg/mL), but prolonged exposure at higher doses (300-500 μg/mL, 10-15 days) showed cytotoxicity to cells (∼95% death) and reproductive toxicity to C. elegans (5-10% reduction in brood size). The origin of toxicity was found to be due to the highly fragmented smaller graphene sheets (<200 nm), while the larger sheets were nontoxic (50-300 μg/mL dose). In contrast, graphene produced with sodium cholate as the mediator has been found to be cytotoxic to these cells at these dosages. We demonstrated the toxicity of liquid phase exfoliated graphene is attributed to highly fragmented fractions or nonbiocompatible exfoliating agents. Thus, low-toxicity graphene/serum suspensions are produced by a facile method in biological media, and this approach may accelerate the much-anticipated development of graphene for biological applications.

Salmonella Enteritidis (SE), Salmonella Typhimurium (ST), and Salmonella Heidelberg (SH) have been responsible for numerous outbreaks associated with the consumption of poultry meat and eggs. Salmonella colonization in chicken is characterized by initial attachment to the cecal epithelial cells (CEC) followed by dissemination to the liver, spleen, and oviduct. Since cecal colonization is critical to Salmonella transmission along the food chain continuum, reducing this intestinal association could potentially decrease poultry meat and egg contamination. Hence, this study investigated the efficacy of Lactobacillus delbreuckii sub species bulgaricus (NRRL B548; LD), Lactobacillus paracasei (DUP-13076; LP), and Lactobacillus rhamnosus (NRRL B442; LR) in reducing SE, ST, and SH colonization in CEC and survival in chicken macrophages. Additionally, their effect on expression of Salmonella virulence genes essential for cecal colonization and survival in macrophages was evaluated. All three probiotics significantly reduced Salmonella adhesion and invasion in CEC and survival in chicken macrophages ( p < 0.05). Further, the probiotic treatment led to a significant reduction in Salmonella virulence gene expression ( p < 0.05). Results of the study indicate that LD, LP, and LR could potentially be used to control SE, ST, and SH colonization in chicken. However, these observations warrant further in vivo validation., Competing Interests: The authors declare no conflict of interest.

Urinary tract infections (UTIs) in the United States result in more than 7 million hospital visits per year. Uropathogenic Escherichia coli (UPEC) is responsible for more than 80% of UTIs. Although antibiotics are the drug of choice to control UTIs, their repeated use has resulted in the emergence of antibiotic-resistant UPEC. Thus, there is a need for effective alternate strategies to control UPEC infections. This study investigated the efficacy of trans-cinnamaldehyde (TC), a food-grade molecule present in cinnamon, in reducing UPEC colonization and pathogenesis in the lower UTI. Female C57BL/6 mice (6-8 weeks old) were fed ad libitum with 0, 0·1, 0·2 and 0·4% TC containing mouse chow for 10 days. Following TC supplementation, animals were experimentally infected with UPEC by transurethral catheterization. Mice were euthanized on days 1, 2 and 4 postinfection, and the bladder, urethra and urine were collected for bacterial enumeration. Prophylactic TC supplementation significantly (P ≤ 0·05) reduced UPEC colonization in the urinary bladder and urethra compared to the control. Results indicate that TC could potentially be used as an oral supplement to control UPEC-associated lower UTIs, however, follow-up clinical trials are warranted., Significance and Impact of the Study: In this study, we have demonstrated that oral supplementation of trans-cinnamaldehyde (TC) reduced uropathogenic Escherichia coli (UPEC)-associated lower urinary tract infection (UTI) in mice. Specifically, in-feed supplementation of TC significantly decreased UPEC populations in the urethra and bladder, thereby reducing the infectious load. These findings are particularly significant given the increase in incidence and prevalence of antibiotic-resistant UTIs. Our study offers new insights into the potential use of natural antimicrobials including TC, the active ingredient in cinnamon, as a nonantibiotic-based natural dietary intervention in the prophylaxis of lower UTIs., (© 2017 The Society for Applied Microbiology.)

Probiotics are nonpathogenic microorganisms that confer a health benefit on the host when administered in adequate amounts. Ample evidence is documented to support the potential application of probiotics for the prevention and treatment of infections. Health benefits of probiotics include prevention of diarrhea, including antibiotic-associated diarrhea and traveler's diarrhea, atopic eczema, dental carries, colorectal cancers, and treatment of inflammatory bowel disease. The cumulative body of scientific evidence that demonstrates the beneficial effects of probiotics on health and disease prevention has made probiotics increasingly important as a part of human nutrition and led to a surge in the demand for probiotics in clinical applications and as functional foods. The ability of probiotics to promote health is attributed to the various beneficial effects exerted by these microorganisms on the host. These include lactose metabolism and food digestion, production of antimicrobial peptides and control of enteric infections, anticarcinogenic properties, immunologic enhancement, enhancement of short-chain fatty acid production, antiatherogenic and cholesterol-lowering attributes, regulatory role in allergy, protection against vaginal or urinary tract infections, increased nutritional value, maintenance of epithelial integrity and barrier, stimulation of repair mechanism in cells, and maintenance and reestablishment of well-balanced indigenous intestinal and respiratory microbial communities. Most of these attributes primarily focus on the effect of probiotic supplementation on the host. Hence, in most cases, it can be concluded that the ability of a probiotic to protect the host from infection is an indirect result of promoting overall health and well-being. However, probiotics also exert a direct effect on invading microorganisms. The direct modes of action resulting in the elimination of pathogens include inhibition of pathogen replication by producing antimicrobial substances like bacteriocins, competition for limiting resources in the host, antitoxin effect, inhibition of virulence, antiadhesive and antiinvasive effects, and competitive exclusion by competition for binding sites or stimulation of epithelial barrier function. Although much has been documented about the ability of probiotics to promote host health, there is limited discussion on the above mentioned effects of probiotics on pathogens. Being in an era of antibiotic resistance, a better understanding of this complex probiotic-pathogen interaction is critical for development of effective strategies to control infections. Therefore, this chapter will focus on the ability of probiotics to directly modulate the infectious nature of pathogens and the underlying mechanisms that mediate these effects., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Acinetobacter baumannii is a major nosocomial pathogen causing human infections with significant mortality rates. In most cases, infections are acquired through exposure to A. baumannii biofilms that persist on contaminated hospital equipment and surfaces. Thus, it is imperative to develop effective measures for controlling A. baumannii biofilms in nosocomial settings. This study investigated the efficacy of octenidine dihydrochloride (OH), a new generation disinfectant for reducing A. baumannii biofilms on polystyrene, stainless steel and catheters. OH at 0.3% (5 mM), 0.6% (10 mM), and 0.9% (15 mM) was effective in significantly inactivating A. baumannii biofilms on all tested surfaces (P < 0.05). Furthermore, OH was equally effective in inactivating biofilms of multidrug resistant and drug susceptible A. baumannii isolates. In addition, confocal imaging revealed the predominance of dead cells in the OH-treated samples in comparison to the control. Further, scanning electron microscopy of biofilms formed on catheters revealed that OH treatment significantly reduced A. baumannii biofilm populations in corroboration with our antibiofilm assay. These data underscore the efficacy of OH in inactivating A. baumannii biofilms, thereby suggesting its potential use as a disinfectant or a catheter lock solution to control A. baumannii infections.

Colorectal cancer, breast cancer and skin cancer are commonly-reported cancer types in the U.S. Although radiation and chemotherapy are routinely used to treat cancer, they produce side effects in patients. Additionally, resistance to chemotherapeutic drugs has been noticed in cancers. Thus, there is a need for effective and safe bioprophylactics and biotherapeutics in cancer therapy. The medicinal value of goat milk has been recognized for centuries and is primarily attributed to three fatty acids, namely capric, caprylic and caproic acids. This research investigates the anticancer property of these fatty acids on human colorectal, skin and mammary gland cancer cells. The cancer cells were treated with various concentrations of fatty acids for 48 h, and cell viability was monitored by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. Additionally, real-time quantitative PCR (RT-qPCR) was performed to elucidate the potential anti-cancer mechanisms of the three fatty acids under investigation. Capric, caprylic and caproic acids reduced cancer cell viability by 70% to 90% (p < 0.05) compared to controls. RT-qPCR data indicated that these natural molecules produced anticancer effects by down-regulating cell cycle regulatory genes and up-regulating genes involved in apoptosis. Future research will validate the anticancer effect of these fatty acids in an appropriate in vivo model.

Cronobacter sakazakii is a foodborne pathogen, which causes a life-threatening form of meningitis, necrotizing colitis and meningoencephalitis in neonates and children. Epidemiological studies implicate dried infant formula as the principal source of C. sakazakii. In this study, we investigated the efficacy of sub-inhibitory concentrations (SIC) of trans-cinnamaldehyde (TC), an ingredient in cinnamon, for reducing C. sakazakii virulence in vitro using cell culture, microscopy and gene expression assays. TC significantly (p ≤ 0.05) suppressed C. sakazakii adhesion to and invasion of human and rat intestinal epithelial cells, and human brain microvascular endothelial cells. In addition, TC inhibited C. sakazakii survival and replication in human macrophages. We also observed that TC reduced the ability of C. sakazakii to cause cell death in rat intestinal cells, by inhibiting nitric oxide production. Results from gene expression studies revealed that TC significantly downregulated the virulence genes critical for motility, host tissue adhesion and invasion, macrophage survival, and LPS (Lipopolysaccharide) synthesis in C. sakazakii. The efficacy of TC in attenuating these major virulence factors in C. sakazakii underscores its potential use in the prevention and/or control of infection caused by this pathogen.

Millions of indwelling devices are implanted in patients every year, and staphylococci (S. aureus, MRSA and vancomycin-resistant S. aureus (VRSA)) are responsible for a majority of infections associated with these devices, thereby leading to treatment failures. Once established, staphylococcal biofilms become resistant to antimicrobial treatment and host response, thereby serving as the etiological agent for recurrent infections. This study investigated the efficacy of octenidine hydrochloride (OH) for inhibiting biofilm synthesis and inactivating fully-formed staphylococcal biofilm on different matrices in the presence and absence of serum protein. Polystyrene plates and stainless steel coupons inoculated with S. aureus, MRSA or VRSA were treated with OH (zero, 0.5, one, 2 mM) at 37 °C for the prevention of biofilm formation. Additionally, the antibiofilm effect of OH (zero, 2.5, five, 10 mM) on fully-formed staphylococcal biofilms on polystyrene plates, stainless steel coupons and urinary catheters was investigated. OH was effective in rapidly inactivating planktonic and biofilm cells of S. aureus, MRSA and VRSA on polystyrene plates, stainless steel coupons and urinary catheters in the presence and absence of serum proteins. The use of two and 10 mM OH completely inactivated S. aureus planktonic cells and biofilm (>6.0 log reduction) on all matrices tested immediately upon exposure. Further, confocal imaging revealed the presence of dead cells and loss in biofilm architecture in the OH-treated samples when compared to intact live biofilm in the control. Results suggest that OH could be applied as an effective antimicrobial to control biofilms of S. aureus, MRSA and VRSA on appropriate hospital surfaces and indwelling devices.

This paper reports an approach to enable rapid concentration and recovery of bacterial cells from aqueous chicken homogenates as a preanalytical step of detection. This approach includes biochemical pretreatment and prefiltration of food samples and development of an automated cell concentration instrument based on cross-flow microfiltration. A polysulfone hollow-fiber membrane module having a nominal pore size of 0.2 μm constitutes the core of the cell concentration instrument. The aqueous chicken homogenate samples were circulated within the cross-flow system achieving 500- to 1,000-fold concentration of inoculated Salmonella enterica serovar Enteritidis and naturally occurring microbiota with 70% recovery of viable cells as determined by plate counting and quantitative PCR (qPCR) within 35 to 45 min. These steps enabled 10 CFU/ml microorganisms in chicken homogenates or 10(2) CFU/g chicken to be quantified. Cleaning and sterilizing the instrument and membrane module by stepwise hydraulic and chemical cleaning (sodium hydroxide and ethanol) enabled reuse of the membrane 15 times before replacement. This approach begins to address the critical need for the food industry for detecting food pathogens within 6 h or less.

Enteric infections account for high morbidity and mortality and are considered to be the fifth leading cause of death at all ages worldwide. Seventy percent of all enteric infections are foodborne. Thus significant efforts have been directed toward the detection, control and prevention of foodborne diseases. Many antimicrobials including antibiotics have been used for their control and prevention. However, probiotics offer a potential alternative intervention strategy owing to their general health beneficial properties and inhibitory effects against foodborne pathogens. Often, antimicrobial probiotic action is non-specific and non-discriminatory or may be ineffective. In such cases, bioengineered probiotics expressing foreign gene products to achieve specific function is highly desirable. In this review we summarize the strategic development of recombinant bioengineered probiotics to control enteric infections, and to examine how scientific advancements in the human microbiome and their immunomodulatory effects help develop such novel and safe bioengineered probiotics.

This study investigated the efficacy of 3 GRAS-status, plant-derived antimicrobials (PDAs), trans-cinnamaldehyde (TC), carvacrol (CR), and β-resorcylic acid (BR) applied as an antimicrobial wash for killing Escherichia coli O157:H7 on apples. "Red delicious" apples inoculated with a 5 strain mixture of E. coli O157:H7 were subjected to washing in sterile deionized water containing 0% PDA (control), 0.15% TC, 0.35% TC, 0.15% CR, 0.30% CR, 0.5% BR, or 1% BR for 1, 3, and 5 min at 23 °C in the presence and absence of 1% soil, and surviving pathogen populations on apples were enumerated at each specified time. All PDAs were more effective in reducing E. coli O157:H7 compared to the water wash treatment (P < 0.05) and reduced the pathogen by 4- to 5-log CFU/apple in 5 min. Chlorine (1%) was the most effective treatment reducing the pathogen on apples to undetectable levels in 1 min (P < 0.05). Moreover, the antimicrobial effect of CR and BR was not affected by the presence of soil, whereas the efficacy of TC and BR was decreased in the presence of soil. Further, no bacteria were detected in the wash solution containing CR and BR; however, E. coli O157:H7 was recovered in the control wash water and treatment solutions containing TC and chlorine, in the presence of 1% soil (P < 0.05). Results suggest that the aforementioned PDAs, especially CR and BR could be used effectively to kill E. coli O157:H7 on apples when used as a wash treatment. Studies on the sensory and quality characteristics of apples treated with PDAs are needed before recommending their usage., (© 2013 Institute of Food Technologists®)

We investigated the efficacy of feed supplemented with caprylic acid (CA), a natural, 8-carbon fatty acid for reducing Salmonella enterica serovar Enteritidis colonization in commercial broiler chickens. In separate 3- and 6-wk trials, 1-d-old straight-run broiler chicks (n = 70 birds/trial) were assigned to a control group (challenged with Salmonella Enteritidis, no CA) and 2 replicates of 0.7 and 1% CA (n = 14 birds/group). Water and feed were provided ad libitum. On d 1, birds were tested for any inherent Salmonella (n = 2 birds/group). For the 3-wk trial, on d 5, birds were challenged with 8 log(10) cfu of Salmonella Enteritidis of a 4-strain mixture by crop gavage, and after 5 d postchallenge, birds (n = 2 birds/group) were euthanized to ensure Salmonella Enteritidis colonization. Caprylic acid was supplemented the last 5 d before tissue collection (n = 10 birds/group). For the 6-wk trial, on d 25, birds were challenged and confirmed for Salmonella Enteritidis colonization. The birds (n = 10 birds/group) were euthanized for tissue samples after CA supplementation for the last 5 d. Caprylic acid at 0.7 or 1% decreased Salmonella Enteritidis populations in cecum, small intestine, cloaca, liver, and spleen in both 3- and 6-wk trials. Body weight of birds did not differ between the groups (P ≥ 0.05). Further, to elucidate a potential antibacterial mechanism of action of CA, we investigated if CA could reduce Salmonella Enteritidis invasion of an avian epithelial cell line and expression of invasion genes hilA and hilD. The cell invasion study revealed that CA reduced invasive abilities of all Salmonella Enteritidis strains by ~80% (P < 0.05). Gene expression studies indicated that CA downregulated (P < 0.001) Salmonella invasion genes hilA and hilD. These results suggest that supplementation of CA through feed could reduce Salmonella Enteritidis colonization in broiler chicken and potentially reduces the pathogen's ability to invade intestinal epithelial cells by downregulating key invasion genes, hilA and hilD.

Listeria monocytogenes (LM) is a major foodborne pathogen causing septicemia, meningitis and death in humans. LM infection is preceded by its attachment to and invasion of human intestinal epithelium followed by systemic spread. The major virulence factors in LM include motility, hemolysin and lecithinase production. Reducing LM attachment to and invasion of host tissue and production of virulence factors could potentially control listeriosis in humans. This study investigated the efficacy of sub-inhibitory concentrations (SICs, concentrations not inhibiting bacterial growth) of three, generally regarded as safe (GRAS)-status, plant-derived antimicrobial compounds in reducing LM attachment to and invasion of human colon adenocarcinoma (Caco-2) and human brain microvascular endothelial cells (HBMEC). Additionally, the effect of these compounds on the aforementioned LM virulence factors was studied. The compounds and their respective SICs used relative to their MICs were trans-cinnamaldehyde (TC 0.50mM, 0.75mM with the MIC of 0.90mM), carvacrol (CR 0.50mM, 0.65mM with the MIC of 0.75mM), and thymol (TY 0.33mM, 0.50mM with the MIC of 0.60mM). All three-plant antimicrobials reduced LM adhesion to and invasion of Caco-2 and HBMEC (p<0.05). The compounds also decreased LM motility, hemolysin production and lecithinase activity (p<0.05). Real-time PCR data revealed that TC, CR, and TY down-regulated the expression of LM virulence genes by >3.0 folds compared to controls (p<0.05). Results suggest that TC, CR, and TY could potentially be used to control LM infection; however, in vivo studies are necessary to validate these results., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The efficacies of trans-cinnamaldehyde (TC) and eugenol (EG) for reducing Salmonella enterica serovar Enteritidis colonization in broiler chickens were investigated. In three experiments for each compound, 1-day-old chicks (n = 75/experiment) were randomly assigned to five treatment groups (n = 15/treatment group): negative control (-ve S. Enteritidis, -ve TC, or EG), compound control (-ve S. Enteritidis, +ve 0.75% [vol/wt] TC or 1% [vol/wt] EG), positive control (+ve S. Enteritidis, -ve TC, or EG), low-dose treatment (+ve S. Enteritidis, +ve 0.5% TC, or 0.75% EG), and high-dose treatment (+ve S. Enteritidis, +ve 0.75% TC, or 1% EG). On day 0, birds were tested for the presence of any inherent Salmonella (n = 5/experiment). On day 8, birds were inoculated with ∼8.0 log(10) CFU S. Enteritidis, and cecal colonization by S. Enteritidis was ascertained (n = 10 chicks/experiment) after 24 h (day 9). Six birds from each treatment group were euthanized on days 7 and 10 after inoculation, and cecal S. Enteritidis numbers were determined. TC at 0.5 or 0.75% and EG at 0.75 or 1% consistently reduced (P < 0.05) S. Enteritidis in the cecum (≥3 log(10) CFU/g) after 10 days of infection in all experiments. Feed intake and body weight were not different for TC treatments (P > 0.05); however, EG supplementation led to significantly lower (P < 0.05) body weights. Follow-up in vitro experiments revealed that the subinhibitory concentrations (SICs, the concentrations that did not inhibit Salmonella growth) of TC and EG reduced the motility and invasive abilities of S. Enteritidis and downregulated expression of the motility genes flhC and motA and invasion genes hilA, hilD, and invF. The results suggest that supplementation with TC and EG through feed can reduce S. Enteritidis colonization in chickens.

Foodborne illness is a serious public health concern. There are over 200 known microbial, chemical, and physical agents that are known to cause foodborne illness. Efforts are made for improved detection, control and prevention of foodborne pathogen in food, and pathogen associated diseases in the host. Several commonly used approaches to control foodborne pathogens include antibiotics, natural antimicrobials, bacteriophages, bacteriocins, ionizing radiations, and heat. In addition, probiotics offer a potential intervention strategy for the prevention and control of foodborne infections. This review focuses on the use of probiotics and bioengineered probiotics to control foodborne pathogens, their antimicrobial actions, and their delivery strategies. Although probiotics have been demonstrated to be effective in antagonizing foodborne pathogens, challenges exist in the characterization and elucidation of underlying molecular mechanisms of action and in the development of potential delivery strategies that could maintain the viability and functionality of the probiotic in the target organ., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Background: Listeria monocytogenes, an intracellular foodborne pathogen, infects immunocompromised hosts. The primary route of transmission is through contaminated food. In the gastrointestinal tract, it traverses the epithelial barrier through intracellular or paracellular routes. Strategies to prevent L. monocytogenes entry can potentially minimize infection in high-risk populations. Listeria adhesion protein (LAP) aids L. monocytogenes in crossing epithelial barriers via the paracellular route. The use of recombinant probiotic bacteria expressing LAP would aid targeted clearance of Listeria from the gut and protect high-risk populations from infection., Methodology/principal Findings: The objective was to investigate the ability of probiotic bacteria or LAP-expressing recombinant probiotic Lactobacillus paracasei (Lbp(LAP)) to prevent L. monocytogenes adhesion, invasion, and transwell-based transepithelial translocation in a Caco-2 cell culture model. Several wild type probiotic bacteria showed strong adhesion to Caco-2 cells but none effectively prevented L. monocytogenes infection. Pre-exposure to Lbp(LAP) for 1, 4, 15, or 24 h significantly (P<0.05) reduced adhesion, invasion, and transepithelial translocation of L. monocytogenes in Caco-2 cells, whereas pre-exposure to parental Lb. paracasei had no significant effect. Similarly, Lbp(LAP) pre-exposure reduced L. monocytogenes translocation by as much as 46% after 24 h. Lbp(LAP) also prevented L. monocytogenes-mediated cell damage and compromise of tight junction integrity. Furthermore, Lbp(LAP) cells reduced L. monocytogenes-mediated cell cytotoxicity by 99.8% after 1 h and 79% after 24 h., Conclusions/significance: Wild type probiotic bacteria were unable to prevent L. monocytogenes infection in vitro. In contrast, Lbp(LAP) blocked adhesion, invasion, and translocation of L. monocytogenes by interacting with host cell receptor Hsp60, thereby protecting cells from infection. These data show promise for the use of recombinant probiotics in preventing L. monocytogenes infection in high-risk populations.

Cronobacter sakazakii is an opportunistic foodborne pathogen that contaminates powdered infant formula, causing a rare but life-threatening infection in neonates and infants. Contaminated powdered infant formula represents the only known source of infection in infants. We previously reported that trans-cinnamaldehyde (TC), an ingredient in cinnamon, inactivated C. sakazakii in powdered infant formula. Although the antimicrobial properties of TC have been well established, only limited information is available on its antimicrobial mechanisms, especially at the molecular level. Therefore, we performed a proteomic analysis of the outer membrane and whole cell proteins from TC-treated C. sakazakii to investigate its potential antimicrobial mechanisms against C. sakazakii. The proteomic data revealed that TC exerts antimicrobial effects by several mechanisms, including disruption of carbohydrate, amino acid, and lipid metabolism. Additionally, TC compromises motility, attachment, and invasion ability and cellular defenses of C. sakazakii against oxidative stress, thereby reducing its virulence. The results of this study suggest that TC could be potentially used for controlling C. sakazakii.

Purpose: Uropathogenic Escherichia coli is the primary bacterium causing urinary tract infection in humans. Attachment and invasion of urinary tract epithelial cells by UPEC is the first critical step in establishing a successful urinary tract infection. We investigated the efficacy of subinhibitory concentrations of trans-cinnamaldehyde to inhibit uropathogenic E. coli attachment and invasion of human uroepithelial cells. We also determined the trans-cinnamaldehyde effect on uropathogenic E. coli genes encoding virulence factors critical for uroepithelial cell bacterial attachment and invasion., Materials and Methods: Polystyrene 24-well plates seeded with uroepithelial cells were inoculated with uropathogenic E. coli (about 6.0 log cfu) and subinhibitory concentrations of trans-cinnamaldehyde (0, 325, 560 and 750 μM), and incubated for 60 minutes at 37C. Uroepithelial cells were washed and lysed to enumerate adhered uropathogenic E. coli populations. For the invasion assay uroepithelial cells were treated with gentamicin after incubation and lysed to enumerate invaded uropathogenic E. coli. Also, the trans-cinnamaldehyde effect on uropathogenic E. coli genes encoding attachment and invasion associated virulence factors was determined by real-time quantitative polymerase chain reaction., Results: Trans-cinnamaldehyde significantly decreased uroepithelial cell attachment and invasion by uropathogenic E. coli (p <0.05). Real-time quantitative polymerase chain reaction revealed that trans-cinnamaldehyde significantly decreased the expression of major genes involved in uropathogenic E. coli attachment and invasion of host tissue (p <0.05). The down-regulating effect of trans-cinnamaldehyde on these genes potentially translated into decreased ability of uropathogenic E. coli to attach and invade bladder cells., Conclusions: Trans-cinnamaldehyde may potentially be used as a safe, effective antimicrobial to control uropathogenic E. coli infection. Followup studies in animal models are warranted., (Copyright © 2011 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.)

Cronobacter sakazakii is an emerging foodborne pathogen transmitted exclusively through contaminated infant formula (IFM), and associated with life-threatening infections in infants. C. sakazakii has the ability to tolerate a variety of environmental stress conditions, including heat stress, acidity, high osmotic pressure, and desiccation. In this study, we investigated the efficacy of a subinhibitory concentration (750 μM) of trans-cinnamaldehyde (TC), an ingredient in cinnamon, for reducing C. sakazakii's tolerance to these environmental stresses. Three strains of TC-treated C. sakazakii were separately subjected to high temperature (50°C, 55°C, and 60°C), acidic pH (3.3), high osmotic pressure (a(w) 0.81), and desiccation. TC (750 μM) substantially (p < 0.05) compromised stress tolerance of C. sakazakii compared to C. sakazakii cells not exposed to TC. Real-time quantitative polymerase chain reaction results revealed that TC significantly (p < 0.05) downregulated C. sakazakii genes critical for stress tolerance and survival, including rpoS, chaperonins, phoP/Q, outer membrane porins, and osmolyte transporter genes. The efficacy of TC in reducing C. sakazakii stress tolerance underscores its potential use for controlling the pathogen by increasing its susceptibility to commonly applied hurdles in food processing.

The efficacy of trans-cinnamaldehyde (TC), an ingredient in cinnamon oil, for inhibiting biofilm synthesis (560 and 750 μM TC) and inactivating mature biofilms (23 and 38 mM TC) of Cronobacter sakazakii was investigated at 24 and 12 °C in the presence and absence of reconstituted infant formula on polystyrene plates, stainless steel coupons, feeding bottle coupons, and enteral feeding tube coupons. Additionally, TC's effect on the expression of genes critical for biofilm formation in C. sakazakii was determined by reverse transcription quantitative PCR. TC inhibited and inactivated C. sakazakii biofilms on all matrices tested at both temperatures. C. sakazakii was reduced by >4.0 and 3.0 log CFU/ml after 96 h of exposure to 38 mM and 750 μM TC, respectively. Reverse transcription quantitative PCR results revealed that TC significantly (P≤0.05) down-regulated biofilm-associated genes in C. sakazakii. TC could potentially be used to control C. sakazakii biofilms on infant formula feeding equipment and preparatory areas., (Copyright ©, International Association for Food Protection)

The efficacy of carvacrol (CAR), trans-cinnamaldehyde (TC), eugenol (EUG) and β-resorcylic acid (BR) as a wash treatment for reducing Salmonella spp. on tomatoes was investigated. Plum tomatoes inoculated with a six-serotype mixture of Salmonella (10⁸CFU) were subjected to washing in sterile deionized water (control) or deionized water containing chlorine (100 ppm), CAR (0.25 and 0.75%), TC (0.5 and 0.75%), EUG (0.25 and 0.75%), or BR (0.75 and 1.0%) for 15 sec, 1 min, and 3 min. The plant molecules were more effective (P<0.05) in reducing Salmonella on tomatoes compared to washing in water and chlorine. Both concentrations of CAR and TC, and 0.75% EUG decreased Salmonella counts on tomatoes by~6.0 log CFU/ml at 1 min. Both concentrations of BR decreased the pathogen on tomatoes to undetectable levels at 3 min of exposure. Washing of tomatoes in deionized water and chlorine for 3 min reduced Salmonella by ca. 2.0 and 4.0 log CFU/ml, respectively. No Salmonella was detected in the wash water containing the plant molecules or chlorine, whereas a substantial population of the pathogen survived in the control wash water. Moreover, none of the dipping treatments had any effect on the red color of tomatoes (P>0.05). Results indicate that CAR, TC, EUG and BR could effectively be used to kill Salmonella on tomatoes, but additional studies on sensory and quality characteristics of tomatoes treated with plant molecules are warranted., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Background: Listeria adhesion protein (LAP) is a housekeeping bifunctional enzyme consisting of N-terminal acetaldehyde dehydrogenase (ALDH) and C-terminal alcohol dehydrogenase (ADH). It aids Listeria monocytogenes in crossing the epithelial barrier through a paracellular route by interacting with its host receptor, heat shock protein 60 (Hsp60). To gain insight into the binding interaction between LAP and Hsp60, LAP subdomain(s) participating in the Hsp60 interaction were investigated., Methods: Using a ModBase structural model, LAP was divided into 4 putative subdomains: the ALDH region contains N1 (Met(1)-Pro(223)) and N2 (Gly(224)-Gly(411)), and the ADH region contains C1 (Gly(412)-Val(648)) and C2 (Pro(649)-Val(866)). Each subdomain was cloned and overexpressed in Escherichia coli and purified. Purified subdomains were used in ligand overlay, immunofluorescence, and bead-based epithelial cell adhesion assays to analyze each domain's affinity toward Hsp60 protein or human ileocecal epithelial HCT-8 cells., Results: The N2 subdomain exhibited the greatest affinity for Hsp60 with a K(D) of 9.50±2.6 nM. The K(D) of full-length LAP (7.2±0.5 nM) to Hsp60 was comparable to the N2 value. Microspheres (1 µm diameter) coated with N2 subdomain showed significantly (P<0.05) higher binding to HCT-8 cells than beads coated with other subdomains and this binding was inhibited when HCT-8 cells were pretreated with anti-Hsp60 antibody to specifically block epithelial Hsp60. Furthermore, HCT-8 cells pretreated with purified N2 subdomain also reduced L. monocytogenes adhesion by about 4 log confirming its involvement in interaction with epithelial cells., Conclusion: These data indicate that the N2 subdomain in the LAP ALDH domain is critical in initiating interaction with mammalian cell receptor Hsp60 providing insight into the molecular mechanism of pathogenesis for the development of potential anti-listerial control strategies.

The effect of trans-cinnamaldehyde (TC) on the inactivation of Escherichia coli O157:H7 in undercooked ground beef patties was investigated. A five-strain mixture of E. coli O157:H7 was inoculated into ground beef (7.0log CFU/g), followed by addition of TC (0, 0.15, and 0.3%). The meat was formed into patties and stored at 4 degrees C for 5 days or at -18 degrees C for 7 days. The patties were cooked to an internal temperature of 60 or 65 degrees C, and E. coli O157:H7 was enumerated. The numbers of E. coli O157:H7 did not decline during storage of patties. However, cooking of patties containing TC significantly reduced (P<0.05) E. coli O157:H7 counts, by >5.0log CFU/g, relative to the reduction in controls cooked to the same temperatures. The D-values at 60 and 65 degrees C of E. coli O157:H7 in TC-treated patties (1.85 and 0.08min, respectively) were significantly lower (P<0.05) than the corresponding D-values for the organism in control patties (2.70 and 0.29min, respectively). TC-treated patties were more color stable and showed significantly lower lipid oxidation (P<0.05) than control samples. TC enhanced the heat sensitivity of E. coli O157:H7 and could potentially be used as an antimicrobial for ensuring pathogen inactivation in undercooked patties. However detailed sensory studies will be necessary to determine the acceptability to consumers of TC in ground beef patties., (Copyright 2010 Elsevier Ltd. All rights reserved.)

Purpose: Urinary tract infections are the most common hospital acquired infections in humans, caused primarily by uropathogenic Escherichia coli. Indwelling urinary catheters for bladder drainage in humans become encrusted with uropathogenic E. coli biofilms that are resistant to common antibiotics, resulting in chronic infections. We studied the efficacy of the cinnamon ingredient trans-cinnamaldehyde (Sigma) for preventing uropathogenic E. coli biofilm. We also determined the efficacy of trans-cinnamaldehyde as an ingredient in catheter lock solution to inactivate preformed uropathogenic E. coli biofilm., Materials and Methods: Polystyrene plates and urinary catheters inoculated with uropathogenic E. coli (5 to 6.0 log cfu) were treated with trans-cinnamaldehyde (0%, 0.1%, 0.25% or 0.5%) at 37C. Catheters with uropathogenic E. coli biofilm were also treated with lock solution containing trans-cinnamaldehyde (0%, 1%, 1.25% or 1.5%). Uropathogenic E. coli biofilm on control and trans-cinnamaldehyde treated plates and catheters was determined on incubation days 0, 1, 3 and 5. Trans-cinnamaldehyde potential cytotoxity, if any, was determined in HTB-4 bladder epithelial cells (ATCC)., Results: At all concentrations trans-cinnamaldehyde effectively prevented uropathogenic E. coli biofilm on plates and catheters. As a constituent in catheter lock solution, it inactivated uropathogenic E. coli biofilm on catheters. Trans-cinnamaldehyde produced no cytotoxic effects on human bladder epithelial cells at the tested concentrations., Conclusions: Results suggest that trans-cinnamaldehyde may be applied as a catheter surface coating or as an ingredient in catheter lock solution to prevent urinary tract infection in humans., (Copyright (c) 2010 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.)

This study investigated the antimicrobial effect of low concentrations of trans-cinnamaldehyde (TC) on Escherichia coli O157:H7 in apple juice and apple cider. A five-strain mixture of E. coli O157:H7 was inoculated into apple juice or cider at approximately 6.0 log CFU/ml, followed by the addition of TC (0%v/v, 0.025%v/v, 0.075%v/v and 0.125%v/v). The inoculated apple juice samples were incubated at 23 degrees C and 4 degrees C for 21 days, whereas the cider samples were stored only at 4 degrees C. The pH of apple juice and cider, and E. coli O157:H7 counts were determined on days 0, 1, 3, 5, 7, 14 and 21. TC was effective (P<0.05) in inactivating E. coli O157:H7 in apple juice and apple cider. At 23 degrees C, 0.125 and 0.075%v/v TC completely inactivated E. coli O157:H7 in apple juice (negative by enrichment) on days 1 and 3, respectively. At 4 degrees C, 0.125 and 0.075%v/v TC decreased the pathogen counts in the juice and cider to undetectable levels on days 3 and 5, respectively. Results indicate that low concentrations of TC could be used as an effective antimicrobial to inactivate E. coli O157:H7 in apple juice and apple cider., (2010 Elsevier B.V. All rights reserved.)

Listeria monocytogenes is a food-borne pathogen capable of forming biofilms and persisting in food processing environments for extended periods of time, thereby potentially contaminating foods. The efficacy of octenidine hydrochloride (OH) for inactivating planktonic cells and preformed biofilms of L. monocytogenes was investigated at 37, 21, 8, and 4 degrees C in the presence and absence of organic matter (rehydrated nonfat dry milk). OH rapidly killed planktonic cells and biofilms of L. monocytogenes at all four temperatures. Moreover, OH was equally effective in killing L. monocytogenes biofilms on polystyrene and stainless steel matrices in the presence and absence of organic matter. The results underscore OH's ability to prevent establishment of L. monocytogenes biofilms by rapidly killing planktonic cells and to eliminate preformed biofilms, thus suggesting that it could be used as a disinfectant to prevent L. monocytogenes from persisting in food processing environments.

Salmonella Enteritidis is a major foodborne pathogen for which chickens serve as reservoir hosts. Reducing Salmonella Enteritidis carriage in chickens would reduce contamination of poultry meat and eggs with this pathogen. We investigated the prophylactic efficacy of feed supplemented with caprylic acid (CA), a natural, generally recognized as safe eight-carbon fatty acid, for reducing Salmonella Enteritidis colonization in chicks. One hundred commercial day-old chicks were randomly divided into five groups of 20 birds each: CA control (no Salmonella Enteritidis, CA), positive control (Salmonella Enteritidis, no CA), negative control (no Salmonella Enteritidis, no CA), and 0.7 or 1% CA. Water and feed were provided ad libitum. On day 8, birds were inoculated with 5.0 log CFU of Salmonella Enteritidis by crop gavage. Six birds from each group were euthanized on days 1, 7, and 10 after challenge, and Salmonella Enteritidis populations in the cecum, small intestine, cloaca, crop, liver, and spleen were enumerated. The study was replicated three times. CA supplementation at 0.7 and 1% consistently decreased Salmonella Enteritidis populations recovered from the treated birds. Salmonella Enteritidis counts in the tissue samples of CA-treated chicks were significantly lower (P < 0.05) than those of control birds on days 7 and 10 after challenge. Feed intake and body weight did not differ between the groups. Histological examination revealed no pathological changes in the cecum and liver of CA-supplemented birds. The results suggest that prophylactic CA supplementation through feed can reduce Salmonella Enteritidis colonization in day-old chicks and may be a useful treatment for reducing Salmonella Enteritidis carriage in chickens.

Enterobacter sakazakii is an emerging pathogen which causes a life-threatening form of meningitis, necrotizing colitis and meningoencephalitis in neonates and children. Epidemiological studies implicate dried infant formula as the principal source of the pathogen. Trans-cinnamaldehyde is a major component of bark extract of cinnamon. It is classified as generally recognized as safe (GRAS) by the U.S. Food and Drug Administration, and is approved for use in food (21 CFR 182.60). The objective of this study was to determine the antibacterial effect of trans-cinnamaldehyde on E. sakazakii in reconstituted infant formula. A 5-strain mixture of E. sakazakii was inoculated into 10 ml samples of reconstituted infant formula (at 6.0 log CFU/ml) containing 0%, 0.15%, 0.3% or 0.5% trans-cinnamaldehyde. The samples were incubated at 37, 23, 8 or 4 degrees C for 0, 6, 10 and 24 h, and the surviving populations of E. sakazakii at each sampling time were enumerated. In addition, potential cytotoxicity of trans-cinnamaldehyde, if any, was determined on human embryonic intestinal cells (INT-407). The treatments containing trans-cinnamaldehyde significantly reduced (P<0.05) the population of E. sakazakii, compared to the controls. Trans-cinnamaldehyde (0.5%) reduced the pathogen to undetectable levels by 4 h of incubation at 37 or 23 degrees C and 10 h of incubation at 8 or 4 degrees C, respectively. Trans-cinnamaldehyde produced no cytotoxic effects on human embryonic intestinal cells at the tested concentrations. Results indicate that trans-cinnamaldehyde could potentially be used to kill E. sakazakii in reconstituted infant formula, however sensory studies are warranted before recommending its use.

The antilisterial activity of monocaprylin (MC) and its combination with acetic acid (AA) on frankfurters was investigated. Each frankfurter was surface inoculated with a three-strain mixture of Listeria monocytogenes to obtain an inoculation level of 4.0 log CFU per frankfurter, and then dipped for 35 s in sterile deionized water (45 or 50 degrees C) containing 1% ethanol (control), 50 mM MC plus 1% ethanol, 1% AA plus 1% ethanol, or 50 mM MC plus 1% AA plus 1% ethanol. Samples were vacuum packaged, stored at 4 degrees C for 77 days, and analyzed for L. monocytogenes. Sensory odor and color of frankfurters were evaluated using a 9-point hedonic scale. Color was also objectively measured using the Minolta Chroma Meter. From day 0 to day 77, population counts of L. monocytogenes on frankfurters dipped in antimicrobial solutions at 50 degrees C were consistently lower than the control counts. Similar results were observed for samples treated at 45 degrees C. However, L. monocytogenes grew readily on control samples at both temperatures. Dipping of frankfurters in antimicrobial solutions (45 or 50 degrees C) significantly reduced (P < 0.05) the populations of L. monocytogenes. After 70 days of storage, L. monocytogenes was completely killed in samples dipped in MC+AA solution at 50 degrees C. The antimicrobial treatments did not affect the odor or color of the samples (P > 0.05). Overall, results indicated that dipping of frankfurters with MC reduced L. monocytogenes, and inclusion of AA further enhanced MC antilisterial activity, without any negative effect on odor or color.

Escherichia coli O157:H7 is an important foodborne pathogen. Cattle serve as one of the major reservoirs of E. coli O157:H7, excreting the pathogen in feces. Environmental persistence of E. coli O157:H7 is critical in its epidemiology on farms, and the pathogen has been isolated from cattle water troughs. Thus, there is a need for an effective method for killing E. coli O157:H7 in cattle drinking water. In this study, the efficacy of sodium caprylate for killing E. coli O157:H7 in cattle drinking water was investigated. A four-strain mixture of E. coli O157:H7 was inoculated (6.0 log CFU/ml) into 100-ml samples of well water containing 0, 75, 100, or 120 mM sodium caprylate. Water samples containing 1% (wt/vol) bovine feces or feed also were included. The samples were incubated at 21 or 8 degrees C for 21 days. Water samples were analyzed for viable E. coli O157:H7 on days 0, 1, 3, 5, and 7 and weekly thereafter. Triplicate samples of each treatment and control were included, and the study was repeated twice. The magnitude of E. coli O157:H7 inactivation in water significantly increased (P < 0.01) with increases in caprylate concentration and storage temperature. At 120 mM, sodium caprylate completely inactivated E. coli O157:H7 in all the samples after 1 to 20 days, depending on the treatments. The presence of feces or feed also had a significant effect (P < 0.01) on the antibacterial property of caprylate; the presence of feces decreased the antibacterial effect, whereas addition of feed enhanced the effect. These results indicate that sodium caprylate is effective in killing E. coli O157:H7 in cattle drinking water, but detailed cattle palatability studies of water containing caprylate are necessary.