Journal of food protection最新文献

Giardia duodenalis is a globally distributed intestinal parasite that commonly infects both humans and animals. G. duodenalis is a species complex, which includes eight assemblages that vary both in genetic structure and host specificity. The prevalence of mixed-assemblage G. duodenalis cysts on food, an understudied infection route for G. duodenalis, remains unknown. In the present study, a method able to detect G. duodenalis mixed-assemblage infections using next-generation amplicon sequencing (NGS) of the beta-giardin gene was applied in combination with the US-FDA’s BAM Chapter 19b protocol for the detection of G. duodenalis from fresh produce to ascertain the limit of detection of G. duodenalis on leafy greens. Ready-to-eat baby Romaine lettuce was inoculated with 5 (n = 5), 20 (n = 10), 100 (n = 10), 200 (n = 10), or 1,000 (n = 10) G. duodenalis cysts of the assemblage B strain H3. Detection of G. duodenalis was successful in 100% of the samples seeded with 1,000, 200, and 100 cysts, in 50% of the samples seeded with 20 cysts, and in none of the samples seeded with 5 cysts. We thus demonstrate robust detection of G. duodenalis on packaged leafy greens using the BAM Chapter 19B method coupled with assemblage-sensitive NGS. This protocol provides a new diagnostic tool useful for both prevalence studies and outbreak investigations involving fresh produce that may assist in better describing the role of G. duodenalis in foodborne illness and in protecting consumers from contaminated fresh produce.

Limu smoked chicken is a traditional Chinese delicacy; however, polycyclic aromatic hydrocarbons (PAHs) are generated during the smoking process. We developed a pyrolysis process for pear wood liquid smoke with minimal PAH generation. Pear wood liquid smoke products were prepared under different pyrolysis conditions in a self-made pyrolysis reactor, and the total phenol, carbonyl compound, total acid, and PAH contents and PAH toxicity risk were evaluated. With increasing temperatures, the toxicity equivalent ΣPAH of the smoke liquid reached 3.004 μg/kg. With increasing particle sizes, the total phenol content reached 1.6 mg/mL; the phenol content was 5.95 mg/mL. With increasing particle sizes, the toxicity equivalent ΣPAHs of the smoke liquor reached 2.441 μg/kg. The optimal parameters for treating pear wood smoke liquid in the thermal reaction device were a pyrolysis temperature of pear wood of >350 °C, particle size of S2, and sucrose content of 8%.

The safety of uncooked fermented, dried sausages relies upon controlled fermentation and drying that inactivates pathogenic bacteria. Current guidelines for the production of fermented sausages by the United States Department of Agriculture (USDA) Food Safety Inspection Services (FSIS) and related research highlight specific safety parameters. The confidence that processing steps, which do not include cooking, inherently mitigate microbial risks, is challenged by the resilience of pathogens in the dry and acidic environments of these food products. The aim of this work was to examine the length of drying required to achieve a target pathogen reduction across a range of sausage diameters. This study investigated the relationship between product diameter and time required to achieve target reductions of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes, as well as the attainment of specific water activity (a_w). The research utilized salami and summer sausage with diameters of 18 mm, 30 mm, 60 mm, 90 mm, and 110 mm. Sausage batter was inoculated with 5 strains each of E. coli O157:H7, L. monocytogenes, and S. enterica. Inoculated sausages were processed with fermentation and drying protocols for each sausage type. Smaller diameter sausages reached both the desired pathogen reduction and target a_w of 0.85 sooner than larger ones. However, the time to achieve the target a_w did not align with the time to achieve the pathogen reduction targets, suggesting that a_w alone is not a reliable indicator of safety. Another finding was larger sausages achieved the target pathogen reduction without reaching the target a_w, suggesting complex relationship between a_w, diameter, and pathogen inactivation. These data support the need for food safety guidelines that consider drying duration, a_w, and pathogen behavior for varying sausage diameters. This research contributes to developing more precise safety protocols for producing dry and semi-dry fermented sausages.

Diarrheal disease is a leading cause of death in children in low- and moderate-income countries. Fresh produce, including fruits and vegetables, may harbor diarrheal disease-causing bacteria including strains of Salmonella enterica and Escherichia coli. This study aimed to determine the prevalence and antibiotic resistance profiles of S. enterica and E. coli isolated from produce samples (n = 207) obtained from retail markets in northern Botswana in Chobe District of Botswana in 2022. Samples were enriched in the appropriate selective media: Brilliant Green Bile Broth for E. coli and Rappaport Vassiliadis Broth for S. enterica. E. coli were confirmed by PCR detecting the phoA gene, and classified as potentially pathogenic through screening for the eae, stx, and stx2 and estIb genes. S. enterica isolates were confirmed using invA primers. Isolates were evaluated for resistance to ampicillin, amoxicillin-clavulanic acid, chloramphenicol, cefotaxime, doxycycline, streptomycin, sulfamethoxazole, and tetracycline antibiotic using the Kirby-Bauer Disk Diffusion method. E. coli was isolated from 15.5% of produce samples (n = 207). The gene eae was detected from 1.5% of samples, while stx1, stx2, and estIb were not detected. Resistance to one or more antibiotics was common (72%) with the majority of the resistant E. coli (n = 32) isolated from fruits (22%) and greens (18%) compared to other types of vegetables. Multidrug resistance (MDR, resistant to 3 or more antibiotics) was identified in 18% of samples. S. enterica was isolated from 3.4% of produce samples (7, n = 207). Resistance was uncommon among the S. enterica isolates (1/7). Overall prevalence of diarrheagenic S. enterica and E. coli was low; however, their presence and that of MDR E. coli in foods commonly consumed raw increases the risk to vulnerable populations. Strategies to reduce contamination of fresh produce and public education on washing and cooking some types of produce may be useful to reduce disease.

Infections of dairy cattle with clade 2.3.4.4b H5N1 highly pathogenic avian influenza virus (HPAIV) were reported in March 2024 in the U.S. and viable virus was detected at high levels in raw milk from infected cows. This study aimed to determine the potential quantities of infectious HPAIV in raw milk in affected states where herds were confirmed positive by USDA for HPAIV (and therefore were not representative of the entire population), and to confirm that the commonly used continuous flow pasteurization using the FDA approved 72 °C (161°F) for 15 s conditions for high−temperature short time (HTST) processing, will inactivate the virus. Double-blinded raw milk samples from bulk storage tanks from farms (n = 275) were collected in four affected states. Samples were screened for influenza A using quantitative real-time RT-PCR (qrRT-PCR) of which 158 (57.5%) were positive and were subsequently quantified in embryonating chicken eggs. Thirty-nine qrRT-PCR positive samples (24.8%) were positive for infectious virus with a median titer of 3.5 log₁₀ 50% egg infectious doses (EID₅₀) per mL. To closely simulate commercial milk pasteurization processing systems, a pilot-scale continuous flow pasteurizer was used to evaluate HPAIV inactivation in artificially contaminated raw milk using the most common legal conditions in the US: 72 °C (161°F) for 15 s. Among all replicates at two flow rates (n = 5 at 0.5 L/min; n = 4 at 1 L/min), no viable virus was detected. A mean reduction of ≥5.8 ± 0.2 log₁₀ EID₅₀/mL occurred during the heating phase where the milk is brought to 72.5 °C before the holding tube. Estimates from heat-transfer analysis support that standard U.S. continuous flow HTST pasteurization parameters will inactivate >12 log₁₀ EID₅₀/mL of HPAIV, which is ∼9 log₁₀ EID₅₀/mL greater than the median quantity of infectious virus detected in raw milk from bulk storage tank samples. These findings demonstrate that the US milk supply is safe when pasteurized.

Antibiotic resistance has become one of the most critical issues in the field of public health in recent years. Exposure to food environment stresses may result in the development of antibiotic resistance in Salmonella. The present study aimed to investigate the simultaneous effects of food-related stresses (osmotic pressure, acid, heat, cold, and freezing stresses) on the antibiotic resistance changes in Salmonella Enteritidis and Salmonella Typhimurium. A factorial design with five factors at two levels was used to evaluate the main and interactive effects of stress factors on the antibiotic resistance of Salmonella serotypes. The changes in the antibiotic resistance of Salmonella serotypes were evaluated using the disc diffusion assay. The results showed that the different stresses had different effects on the antibiotic resistance of Salmonella serotypes. The freezing time and osmotic stresses had the most significant effects on the antibiotic resistance (P < 0.05). S. Enteritidis showed the slightest changes after exposure to stresses. The results also showed that a low level (24 h) of freezing time decreased the antibiotic resistance, but at a high level (96 h) increased it. The results emphasized that food processing and storage conditions should be considered as crucial factors in developing antibiotic resistance in Salmonella.

The need for a sensitive molecular method to detect specific species of insect contaminants in food products remains a significant challenge in the food industry. This study evaluated the detection limit of a multiplex end-point PCR assay for detecting insects in food. The assay amplifies two fragments of the cytochrome oxidase subunit I gene (COI-Fa and COI-Fb) and one fragment of the protein-coding wingless (wg) gene found in insects. Five insect species, comprising three vectors of foodborne pathogens (the housefly, Musca domestica, the American cockroach, Periplaneta americana, and the pharaoh ant, Monomorium pharaonis) and two storage insect pests (the red flour beetle, Tribolium castaneum and the Indian meal moth, Plodia interpunctella), were spiked separately and in combination at levels of 1, 0.1, 0.01, and 0.001% in whole wheat flour. At spike levels greater than 0.01%, amplicon bands of expected sizes were seen in 100% of samples containing fragments from distinct insect species. At least 25% of spiked samples at the lowest spike level had amplicon bands, except for samples spiked with M. domestica. Results showed an 18.9% probability (with 11.3% and 30% lower and upper confidence limits, respectively) of detecting insect fragments at the lowest spike level (0.001%, corresponding to 3–22 fragments), which is far below the FDA’s regulatory level of less than 75 fragments per 50 g of wheat flour. The intensity of amplicon bands in the gel images was higher at higher spike levels. However, this method is not quantitative enough to extrapolate the intensity of the amplicon bands to the number of insect fragments present in a sample. This multiplex assay was also evaluated in a variety of market food samples derived from plants and animals, showing its potential use in various food types. Overall, the sensitivity and specificity of this molecular approach suggest that it could be used in the future as a screening tool for detecting insect contaminants in food.

Poultry-associated salmonellosis results in significant costs to poultry producers and consumers. Given the vertically integrated nature of the United States poultry industry, a better understanding of Salmonella ecology throughout all levels of poultry production is essential. One nexus point is the hatchery, where eggs from multiple broiler breeder farms are incubated and hatched, with the chicks being sent to numerous farms; therefore, the hatchery represents an ideal area to understand preharvest Salmonella ecology and flow. To achieve this, a commercial broiler hatchery was biomapped, focusing on Salmonella prevalence and serotype diversity among four major sample type categories (Air, Egg, Water, Facility) across five different places in the prehatch, hatch, and posthatch areas. Following two sets of eggs from broiler breeder farms over two production days, the overall Salmonella prevalence was 26% (48/184). Of the positive samples, the highest prevalence was observed in swabs taken from the floor drains in the facility and transport truck (56%), as well as in the hatch and posthatch hatchery areas (50%). Kentucky (n = 17), Gaminara (n = 12), and Alachua (n = 11) were the dominant Salmonella serotypes, with serotypes of greatest outbreak concern from chickens (Enteritidis) representing only 6.25% (3/48) of all recovered Salmonella isolates. The posthatch transport area, including the underfloor reservoirs of the transport trucks, not only harbored Enteritidis but also the enrichment broths from these Salmonella-positive samples also possessed sequences matching the commercial live-attenuated vaccine Typhimurium strain according to CRISPR SeroSeq analyses. These findings highlight the complex diversity of commercial hatchery Salmonella populations, including identifying facility floor drains and transport trucks as potentially important critical control points for hatchery managers to focus their Salmonella mitigation efforts to reduce loads and serotypes entering live production farms.

Biological soil amendments of animal origin (BSAAOs) are widely used in urban agriculture to improve soil quality. Although BSAAO use is regulated due to risks for introducing foodborne pathogens, effects on antimicrobial-resistant (AMR) bacteria are not well established. Here, we aimed to explore the impacts of BSAAOs on levels of resident AMR bacteria in leafy vegetable production environments (i.e., kale, lettuce, chard, cabbage) across urban farms and community gardens in the greater Washington D.C. area (n = 7 sites). Leaf tissue (LT), root zone soil (RZS; amended soil in crop beds), and bulk soil (BS; site perimeter) were collected and analyzed for concentrations of total heterotrophic bacteria (THB), ampicillin (Amp) or tetracycline (Tet) resistant THB, and coliforms. As expected, amended plots harbored significantly higher concentrations of THB than bulk soil (P < 0.001). The increases in total bacteria associated with reduced fractions of Tet-resistant bacteria (P = 0.008), as well as case-specific trends for reduced fractions of Amp-resistant bacteria and coliforms. Site-to-site variation in concentrations of AMR bacteria in soil and vegetable samples reflected differences in land history and crop management, while within-site variation was associated with specific amendment sources, as well as vegetable type and cultivar. Representative isolates of the AMR bacteria and coliforms were further screened for multidrug resistance (MDR) phenotypes, and a high frequency was observed for the former. In amended soils, as the soil pH (range 6.56–7.80) positively correlated with the fraction of Tet-resistant bacteria (rho = 0.529; P < 0.001), crop management strategies targeting pH may have applications to control related risks. Overall, our findings demonstrate that soil amendments promote soil bacteria concentrations and have important implications for limiting the spread of AMR bacteria, at least in the urban landscape.

The objective of this study was to reveal the antibacterial mode of action of garviecin LG34 against S. aureus CICC 21600 and L. monocytogenes CICC 21633 and measure the inhibitions on these two foodborne pathogenic bacteria in milk. Antibacterial mechanism of garviecin LG34 was ascertained by its effect on the efflux of Potassium (K⁺) ions, extracellular electrical conductivity, UV-absorbing substances, potential across the membrane (ΔΨ), and cell permeability. The inhibition of garviecin LG34 against S. aureus CICC 21600 and L. monocytogenes CICC 21600 in milk was studied by viable counting method. Supplementation with 160 AU/ml of garviecin LG34 had a bactericidal effect on S. aureus CICC 21600 and L. monocytogenes CICC 21633. A total of 80, 160, and 320 AU/ml of garviecin LG34 resulted in the effusion of potassium ion and UV-absorbing substances, the leakage of cellular electrolytes, and the dissipation of electrical potential across the membrane of these two food-borne bacteria and showed a dose-dependent. Moreover, the increase in cell permeability of both strains was observed by flow cytometer after cells treated with 160 AU/ml of garviecin LG34. Garviecin LG34 significantly inhibited the growth of these two food-borne bacteria in milk, especially in skimmed milk. Garviecin LG34 could cause pore formation, intracellular materials release, and permeability increase of S. aureus CICC 21600 and L. monocytogenes CICC 21633, and could be applied to milk as bio-preservative.