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There is an increasing need to develop alternative antimicrobials to replace currently used antibiotics. Phytochemicals, such as essential oils, have garnered significant attention in recent years as potential antimicrobials. However, the mechanisms underlying their bactericidal activities are not yet fully understood. In this study, we investigated the bactericidal activity of eugenol oil against Salmonella enterica serovar Typhimurium (S. Typhimurium) to elucidate its mechanism of action. We hypothesized that eugenol exerts its bactericidal effects through the production of reactive oxygen species (ROS), which ultimately leads to cell death. The result of this study demonstrated that the bactericidal activity of eugenol against S. Typhimurium was significantly (p < 0.05) mitigated by thiourea (ROS scavenger) or iron chelator 2,2′-dipyridyl, supporting the hypothesis. This finding contributes to a better understanding of the killing mechanism by eugenol oil. [ABSTRACT FROM AUTHOR]

Within the realm of poultry feed mill operations, the persistent concern over microbial feed quality necessitates the establishment of a robust baseline for enhancing and sustaining the standards of commercial feeds. This dual-phase investigation, comprising Parts I, was previously published, and the current study presented here as Part II aimed to illuminate this baseline using 16S rRNA gene sequencing. In Part II, nine distinct commercial poultry feeds formulated as starters, growers, starter/growers, or supplements, the selected feeds underwent genomic DNA extraction, amplification with custom dual-indexed primers, and subsequent Illumina MiSeq sequencing. Through data analysis in QIIME2-2021.4 and R Studio, the study unveils alpha (Kruskal–Wallis) and beta (ANOSIM) diversity, taxonomic differences (ANCOM), and core microbiomes (core_members), deeming main and pairwise effects statistically significant at p < 0.05 and Q < 0.05. Notably, the investigation identified 30% common core microbial members across the nine feed types, shedding light on potential foodborne poultry pathogens such as Helicobacter and Campylobacter. Probiotic-associated feeds exhibited distinct microbial communities, emphasizing the need to explore their impact on the early poultry gastrointestinal tract (GIT) further. [ABSTRACT FROM AUTHOR]

In this study, we explore how transformer models, which are known for their attention mechanisms, can improve pathogen prediction in pastured poultry farming. By combining farm management practices with microbiome data, our model outperforms traditional prediction methods in terms of the F1 score—an evaluation metric for model performance—thus fulfilling an essential need in predictive microbiology. Additionally, the emphasis is on making our model's predictions explainable. We introduce a novel approach for identifying feature importance using the model's attention matrix and the PageRank algorithm, offering insights that enhance our comprehension of established techniques such as DeepLIFT. Our results showcase the efficacy of transformer models in pathogen prediction for food safety and mark a noteworthy contribution to the progress of explainable AI within the biomedical sciences. This study sheds light on the impact of effective farm management practices and highlights the importance of technological advancements in ensuring food safety. [ABSTRACT FROM AUTHOR]

Food additives are employed in the food industry to enhance the color, smell, and taste of foods, increase nutritional value, boost processing efficiency, and extend shelf life. Consumers are beginning to prioritize food ingredients that they perceive as supporting a healthy lifestyle, emphasizing ingredients they deem acceptable as alternative or "clean-label" ingredients. Ready-to-eat (RTE) meat products can be contaminated with pathogens and spoilage microorganisms after the cooking step, contributing to food spoilage losses and increasing the risk to consumers for foodborne illnesses. More recently, consumers have advocated for no artificial additives or preservatives, which has led to a search for antimicrobials that meet these demands but do not lessen the safety or quality of RTE meats. Lactates and diacetates are used almost universally to extend the shelf life of RTE meats by reducing spoilage organisms and preventing the outgrowth of the foodborne pathogen Listeria monocytogenes. These antimicrobials applied to RTE meats tend to be broad-spectrum in their activities, thus affecting overall microbial ecology. It is to the food processing industry's advantage to target spoilage organisms and pathogens specifically. [ABSTRACT FROM AUTHOR]

Campylobacter growth kinetic parameters can be used to refine the sensitivity and efficiency of microbial growth-based methods. Therefore, the aim of this study was to construct growth curves for C. jejuni, C. coli, and C. lari in pure culture and calculate growth kinetics for each Campylobacter species in the same environmental conditions. Campylobacter jejuni, C. coli and C. lari were grown over 48 h and inoculated into 15 mL Hungate tubes (N = 3 trials per species; 5 biological replicates per trial; 3 species; 1 strain per species). Absorbance measurements were taken in 45 min intervals over 24 h. Optical density readings were plotted versus time to calculate growth kinetic parameters. C. jejuni exhibited the longest lag phase (p < 0.001) at 15 h 20 min ± 30 min, versus C. coli at 11 h 15 min ± 17 min, and C. lari at 9 h 27 min ± 15 min. The exponential phase duration was no longer than 5 h for all species, and doubling times were all less than 1h 30 min. The variation in growth kinetics for the three species of Campylobacter illustrates the importance of determining individual Campylobacter spp. growth responses for optimizing detection based on low bacterial levels. This study provides kinetics and estimates to define enrichment times necessary for low concentration Campylobacter detection. [ABSTRACT FROM AUTHOR]

Listeria continues to be a persistent foodborne pathogen that is responsible for human cases of listeriosis when contaminated food products are consumed. Human subjects considered to be most susceptible include the elderly, immunocompromised, and pregnant women. Listeria is characterized as a saprophytic organism with the capability of responding and adapting to constantly changing environments because it possesses multiple stress response mechanisms to overcome varying temperatures, salt concentrations, and pH, among others. Primary foods and food products associated with listeriosis include dairy products and ready-to-eat meats such as turkey products. Historically, chicken eggs have not been identified as a primary source of Listeria, but the potential for contamination during egg production and processing does exist. Listeria species have been isolated from egg-processing plant equipment and are presumed to occur in egg-processing plant environments. Whether Listeria is consistently disseminated onto eggs beyond the egg-processing plant is a risk factor that remains to be determined. However, research has been conducted over the years to develop egg wash solutions that generate combinations of pH and other properties that would be considered inhibitory to Listeria. Even less is known regarding the association of Listeria with alternative egg production systems, but Listeria has been isolated from pasture flock broilers, so it is conceivable, given the nature of the outdoor environments, that layer birds under these conditions would also be exposed to Listeria and their eggs become contaminated. This review focuses on the possibility of Listeria occurring in conventional and alternative egg-laying production and processing systems. [ABSTRACT FROM AUTHOR]

Campylobacter has consistently posed a food safety issue in broiler meat. This study aimed to create a quantitative microbial risk assessment model from retail to consumption, designed to evaluate the seasonal risk of campylobacteriosis associated with broiler meat consumption in the United States. To achieve this, data was gathered to build distributions that would enable us to predict the growth of Campylobacter during various stages such as retail storage, transit, and home storage. The model also included potential fluctuations in concentration during food preparation and potential cross-contamination scenarios. A Monte Carlo simulation with 100,000 iterations was used to estimate the risk of infection per serving and the number of infections in the United States by season. In the summer, chicken meat was estimated to have a median risk of infection per serving of 9.22 × 10−7 and cause an average of about 27,058,680 infections. During the winter months, the median risk of infection per serving was estimated to be 4.06 × 10−7 and cause an average of about 12,085,638 infections. The risk assessment model provides information about the risk of broiler meat to public health by season. These results will help understand the most important steps to reduce the food safety risks from contaminated chicken products. [ABSTRACT FROM AUTHOR]

Campylobacter is a major foodborne pathogen with over a million United States cases a year and is typically acquired through the consumption of poultry products. The common occurrence of Campylobacter as a member of the poultry gastrointestinal tract microbial community remains a challenge for optimizing intervention strategies. Simultaneously, increasing demand for antibiotic-free products has led to the development of several alternative control measures both at the farm and in processing operations. Bacteriophages administered to reduce foodborne pathogens are one of the alternatives that have received renewed interest. Campylobacter phages have been isolated from both conventionally and organically raised poultry. Isolated and cultivated Campylobacter bacteriophages have been used as an intervention in live birds to target colonized Campylobacter in the gastrointestinal tract. Application of Campylobacter phages to poultry carcasses has also been explored as a strategy to reduce Campylobacter levels during poultry processing. This review will focus on the biology and ecology of Campylobacter bacteriophages in poultry production followed by discussion on current and potential applications as an intervention strategy to reduce Campylobacter occurrence in poultry production. [ABSTRACT FROM AUTHOR]

Alternative poultry production systems consisting of free-range or pasture flock raised poultry continues to increase in popularity. Based on the perceived benefits of poultry products generated from these alternative poultry production systems, they have commercial appeal to consumers. Several factors impact the health and well being of birds raised and maintained in these types of production systems. Exposure to foodborne pathogens and potential for colonization in the gastrointestinal tract has to be considered with these types of production systems. The gastrointestinal tract microbial composition and function of birds grown and maintained in alternative poultry operations may differ depending on diets, breed, and age of bird. Dietary variety and foraging behavior are potential influential factors on bird nutrition. The gastrointestinal tract microbiomes of birds raised under alternative poultry production systems are now being characterized with next-generation sequencing to identify individual microbial members and assess the impact of different factors on the diversity of microbial populations. In this review, the gastrointestinal tract microbiota contributions to free-range or pasture-raised broiler and egg layer production systems, subsequent applications, and potential future directions will be discussed. [ABSTRACT FROM AUTHOR]

Broiler (Gallus gallus domesticus) litter is subject to ammonia (NH3) volatilization losses. Previous work has shown that the addition of gypsum to broiler litter can increase nitrogen mineralization and decrease NH3 losses due to a decrease in pH, but the mechanisms responsible for these effects are not well understood. Therefore, three laboratory studies were conducted to evaluate the effect of gypsum addition to broiler litter on (i) urease activity at three water contents, (ii) calcium carbonate precipitation, and (iii) pH. The addition of gypsum to broiler litter increased ammonium concentrations (p < 0.0033) and decreased litter pH by 0.43 to 0.49 pH units after 5 d (p < 0.0001); however, the rate of urea hydrolysis in treated litter only increased on Day 0 for broiler litter with low (0.29 g H2O g-1) and high (0.69 g H2O g-1) water contents, and on Day 3 for litter with medium (0.40 g H2O g-1) water content (p < 0.05). Amending broiler litter with gypsum also caused an immediate decrease in litter pH (0.22 pH units) due to the precipitation of calcium carbonate (CaCO3) from gypsum-derived calcium and litter bicarbonate. Furthermore, as urea was hydrolyzed, more urea-derived carbon precipitated as CaCO3 in gypsum-treated litter than in untreated litter (p < 0.001). These results indicate that amending broiler litter with gypsum favors the precipitation of CaCO3, which buffers against increases in litter pH that are known to facilitate NH3 volatilization. [ABSTRACT FROM AUTHOR]

The use of antibiotics in agroecosystems has been implicated in the rise in antibiotic resistance (AR), which can affect environmental, animal, and human health. To determine the environmental impact of antibiotic use in agroecosystems, appropriate background levels of AR in agricultural environments in the absence of antibiotic application must be determined. Therefore, to determine background levels of AR in broiler production, four target microbes (Escherichia coli, Salmonella, Campylobacter, and Listeria) were isolated from 15 all-natural, antibiotic-free, pastureraised broiler flocks from six farms within the southeastern United States. The AR profiles of these isolates were characterized using the CDC National Antimicrobial Resistance Monitoring System for Enteric Bacteria (NARMS), and these resistance patterns were compared across target microbes and farms and throughout the life cycle of the flocks along the farm-to-fork continuum. Antibiotic resistances were most prevalent in Listeria and Salmonella and least prevalent in Campylobacter. Although E. coli and Salmonella were isolated from the same farms and characterized using the same NARMS plates, they exhibited distinct AR profiles, with Salmonella demonstrating clear farmspecific resistance patterns. Multidrug resistance rates (three or more antibiotics), in order of prevalence, were Listeria (63.9%), Salmonella (36.0%), E. coli (12.7%), and Campylobacter (1.4%). The results of this study demonstrate the variability in background AR among major food safety-related microbes, even when isolated from similar production and processing samples from the same farms, and indicate the need for the proper design of future broiler production studies to account for this highly dynamic background AR. [ABSTRACT FROM AUTHOR]

Although historically, antibiotic resistance has occurred naturally in environmental bacteria, many questions remain regarding the specifics of how humans and animals contribute to the development and spread of antibiotic resistance in agroecosystems. Additional research is necessary to completely understand the potential risks to human, animal, and ecological health in systems altered by antibiotic-resistance-related contamination. At present, analyzing and interpreting the effects of human and animal inputs on antibiotic resistance in agroecosystems is difficult, since standard research terminology and protocols do not exist for studying background and baseline levels of resistance in the environment. To improve the state of science in antibiotic-resistance-related research in agroecosystems, researchers are encouraged to incorporate baseline data within the study system and background data from outside the study system to normalize the study data and determine the potential impact of antibiotic-resistance-related determinants on a specific agroecosystem. Therefore, the aims of this review were to (i) present standard definitions for commonly used terms in environmental antibiotic resistance research and (ii) illustrate the need for research standards (normalization) within and between studies of antibiotic resistance in agroecosystems. To foster synergy among antibiotic resistance researchers, a new surveillance and decision-making tool is proposed to assist researchers in determining the most relevant and important antibiotic-resistance-related targets to focus on in their given agroecosystems. Incorporation of these components within antibiotic-resistance-related studies should allow for a more comprehensive and accurate picture of the current and future states of antibiotic resistance in the environment. [ABSTRACT FROM AUTHOR]

The article discusses the study which examines the biological mineralization of organic nitrogen (ON) in acidified poultry litters based on the population shift from bacteria to fungi. It implies that dry acids delays microbial uric or ammonia emissions, however, the effect of dry acids on biological mineralization of ON has not been established. The study indicates that ON mineralization has resulted in a 2-to 4-week delay of microbial and physiochemical transformations compared to untreated poultry manure.

The article focuses on a study which measured the temporal changes in total bacterial and fungal populations and urease-producing microorganisms in nontreated litter or litter treated with 10% alum. To target the bacterial 16S ribosomal ribonucleic (rRNA) gene, the fungal 18S rRNA gene, or the urease gene of bacterial and fungal ammonia producers in the poultry litter incubation study, quantitative real-time polymerase chain reaction was used. The study found that nontreated poultry litter had relatively high total and ureolytic bacterial population. It also found that alum treatment reduced the total bacterial population by 50% and bacterial urease producers by 90% within 4 weeks.

Skatole is one of the most malodorous compounds produced from the anaerobic degradation of animal waste. Little is known about the biochemistry of skatole production, the phylogeny of skatole-producing microorganisms or the conditions that favor their growth. These deficiencies hamper attempts to reduce skatole production. Our goals were to enrich for skatole producers in swine lagoon slurry (SLS) and evaluate the resulting microbial community structure using denaturing gradient gel electrophoresis (DGGE) and 16S rRNA gene sequence analysis. Skatole producers were enriched by incubating dilutions of SLS with 100 μM indole-3-acetic acid (IAA). GC-MS was used to measure skatole production in the slurries after 0, 7 and 17 days' incubation. Based on most probable number analysis, skatole producers increased 100-fold in SLS samples supplemented with IAA. Based on DGGE fingerprint patterns from day 0, 7 and 17 treatments with high, mid or low levels of skatole production, changes in the SLS population occurred as skatole production increased. Changes in the bacterial community fingerprints were associated with an increase in the low-GC gram-positive and Bacteroides groups. Results from this study provides valuable new information concerning the organisms responsible for production of this odorant, a necessary first step towards controlling skatole production. [ABSTRACT FROM AUTHOR]

Adenine and thymine homopolymer strings of at least 8 nucleotides (AT 8+mers) were characterized in Salmonella enterica subspecies I. The motif differed between other taxonomic classes but not between Salmonella enterica serovars. The motif in plasmids was possibly associated with serovar. Approximately 12.3% of the S. enterica motif loci had mutations. Mutability of AT 8+mers suggests that genomes undergo frequent repair to maintain optimal gene content, and that the motif facilitates self-recognition; in addition, serovar diversity is associated with plasmid content. A theory that genome regeneration accounts for both persistence of predominant Salmonella serovars and serovar diversity provides a new framework for investigating root causes of foodborne illness. [ABSTRACT FROM AUTHOR]

A major concern of the broiler industry is the volatilization of ammonia (NH3) from the mixture of bedding material and broiler excretion that covers the floor of broiler houses. Gypsum has been proposed as a litter amendment to reduce NH3 volatilization, but reports of NH3 abatement vary among studies and the mechanism responsible for decreasing NH3 volatilization is not well understood. The goal of this study was to evaluate the effect of adding 20 or 40% flue-gas desulfurization gypsum (FGDG) to broiler litter on pH, electrical conductivity (EC), water potential, urea-degrading bacteria abundance, NH3 and carbon dioxide (CO2) evolution, and nitrogen (N) mineralization in several 21-d experiments. The addition of FGDG to broiler litter increased EC by 24 to 33% (P < 0.0001), decreased urea-degrading bacteria by 48 to 57% (P = 0.0001) and increased N mineralization by 10 to 11% (P = 0.0001) as compared to litters not amended with FGDG. Furthermore, the addition of FGDG to broiler litter decreased NH3 volatilization by 18 to 28% (P < 0.0001), potentially resulting from the significantly lower litter pH values compared to un-amended litter (P < 0.0001). Findings of this study indicate that amending broiler litter with 20% FGDG can decrease NH3 volatilization and increase the fertlizer value of broiler litter. [ABSTRACT FROM AUTHOR]

Fostering a "balanced" gut microbiome through the administration of beneficial microbes that can competitively exclude pathogens has gained a lot of attention and use in human and animal medicine. However, little is known about how microbes affect the horizontal gene transfer of antimicrobial resistance (AMR). To shed more light on this question, we challenged neonatal broiler chicks raised on reused broiler chicken litter--a complex environment made up of decomposing pine shavings, feces, uric acid, feathers, and feed--with Salmonella enterica serovar Heidelberg (S. Heidelberg), a model pathogen. Neonatal chicks challenged with S. Heidelberg and raised on reused litter were more resistant to S. Heidelberg cecal colonization than chicks grown on fresh litter. Furthermore, chicks grown on reused litter were at a lower risk of colonization with S. Heidelberg strains that encoded AMR on IncI1 plasmids. We used 16S rRNA gene sequencing and shotgun metagenomics to show that the major difference between chicks grown on fresh litter and those grown on reused litter was the microbiome harbored in the litter and ceca. The microbiome of reused litter samples was more uniform and enriched in functional pathways related to the biosynthesis of organic and antimicrobial molecules than that in fresh litter samples. We found that Escherichia coli was the main reservoir of plasmids encoding AMR and that the IncI1 plasmid was maintained at a significantly lower copy per cell in reused litter compared to fresh litter. These findings support the notion that commensal bacteria play an integral role in the horizontal transfer of plasmids encoding AMR to pathogens like Salmonella. [ABSTRACT FROM AUTHOR]

Themicrobiota of the live production environment can directly shape the gastrointestinal microbiome of chickens and indirectly influence the health of birds. Therefore, numerous studies have attempted to characterize the microbial communities in litter and chicken feces from commercial poultry houses, but many treat a poultry house as a single sampling unit. Unfortunately, a poultry house has distinct areas (e.g., waterer/feeder, near fans, cooling pads), so effective sampling strategies need to be developed to account for this heterogeneity, especially when costlier microbiomic analyses are used to assess whole-house microbial diversity. Therefore, the goals of this study were to assess the spatial variability of the poultry litter and fecal microbiomes from distinct areas within a poultry house and to compare composite "whole-house" microbiomes pooled from all house areas, either (1) physically prior to DNA extraction or (2) combined in silico after sample processing (during DNA sequence analysis). No significant differences in α-diversity metrics (richness, diversity, evenness) were observed for fecal or litter microbiomes recovered from the different areas of the house, but β-diversity (litter only) and genus-level Operational Taxonomic Units (OTU) relative abundances (for fecal and litter) were found to be significantly different based on sampling area within the house. Additionally, the sample pooling method produced distinct composite fecal microbiomic profiles, but the litter microbiomes were unaffected. These results indicate that sample type, sampling area, and sample pooling method need to be carefully considered when determining appropriate sampling strategies for generating representative composite whole-house microbiomes for future microbiological-based live production studies. [ABSTRACT FROM AUTHOR]