Escherichia coli O157:H7-adulterated food products are associated with disease outbreaks in humans. Although cattle feces are a source for E. coli O157:H7 contamination, it is unclear if human-associated outbreak isolates differentially colonize and shed in the feces of cattle from that of non-outbreak isolates. It is also unclear if phenotypes, such as biofilm formation, cell attachment, or toxin production, differentiate environmental E. coli O157:H7 isolates from those associated with human illness. The objective of this study was to compare the genotypes and phenotypes of a diverse set of E. coli O157:H7 isolates, with the intent of identifying differences that could inform cattle colonization and fecal shedding, along with virulence potential in humans. Isolates differed in attachment phenotypes on human Caco-2 cells and bovine-derived recto-anal junction squamous epithelial cells, with curli having a strong impact on attachment to the human-derived cell line. The prototypical E. coli O157 isolate EDL933 had the greatest expression of the adhesin gene iha, yet it had decreased expression of the virulence genes stx2, eae, and ehxA compared the lineage I/II isolates RM6067W and/or FRIK1989. Strong or weak biofilm production was not associated with significant differences in cattle colonization or shedding, suggesting biofilms may not play a major role in cattle colonization. No significant differences in cattle colonization and fecal shedding were detected, despite genomic and in vitro phenotypic differences. The outbreak isolate associated with the greatest incidence of hemolytic uremic syndrome, RM6067W, induced the greatest Vero cell cytotoxicity and had the greatest stx2 gene expression.
OBJECTIVE: Foodborne illness has major impacts on global health and imposes financial hardships on food industries. Escherichia coli serotype O157:H7 is associated with foodborne illness. Cattle feces are a source of E. coli O157:H7, and routine surveillance has led to an abundance of E. coli O157:H7 genomic data. The relationship between E. coli O157:H7 genome and phenotype is not clearly discerned for cattle colonization/shedding and improved understanding could lead to additional strategies to limit E. coli O157:H7 in the food chain. The goal of the research was to evaluate genomic and phenotypic attributes of E. coli O157:H7 associated with cattle colonization and shedding, environmental persistence, and human illness. Our results indicate variations in biofilm formation and in vitro cellular adherence was not associated with differences in cattle colonization or shedding. Overall, processes involved in cattle colonization and various phenotypes in relation to genotype are complex and remain not well understood.

This forum explores the hypothesis that plants serve as a hub for the evolution of generalist Salmonella strains, and can be traced back to the Neolithic era when agriculture emerged. It suggests a unique interconnection among humans, plants, animals, and Salmonella, potentially driving the evolution of the pathogen on farms.

BACKGROUND: Acute Infectious Diarrhea (AID) and the short- and long-term complications associated with it are major causes of hospitalization worldwide. In Italy, due to a lack of robust surveillance programs, only limited data has been collected on their prevalence and circulation. This study aims to evaluate the resistance pattern of enteric pathogens and their epidemiological trends over a six-year period.
METHODS: This cross-sectional retrospective study was conducted from January 2018 to December 2023. Stool samples were analyzed during routine diagnosis with culture methods, syndromic molecular tests, and enzyme immunoassay.
RESULTS: Bacteria were the most isolated enteric pathogens (62.2%), followed by fungi (29.0%), viruses (8.2%), and parasites (0.6%). Most bacteria were isolated from outpatients (29.5%) and from patients in the Oncology ward (26.2%). The most prevalent target was EPEC (11.1%), followed by C. difficile toxin A/B-producing strains (8.3%), C. jejuni (2.5%), and S. enterica, (1%.). Norovirus and Candida spp. were the most prevalent in pediatric patients (6.5% and 39.6%, respectively). In the last years, enteric pathogens have been a frequent cause of infections characterized by a problematic resistance to common antimicrobials. In our study, S. enterica showed resistance to amikacin, gentamicin, ampicillin, levofloxacin, and ciprofloxacin. C. jejuni was susceptible to all tested drugs.
CONCLUSIONS: Timely notification of gastroenteric infections is crucial in identifying potential outbreak sources and ensuring strict adherence to food safety and hygiene practices, so as to protect the most vulnerable populations. The present study offers insights into the epidemiological characteristics and the antibiotic susceptibility of the main enteric AID pathogens in order to implement infection control measures in health care settings.

Introduction An acute gastrointestinal illness outbreak was reported in a higher educational institution among students and faculties in East Sikkim, India, from January to February 2023. The investigation was conducted to identify the source of the infection and causative pathogens and prevent the spread of the outbreak. Methods We defined a case as three or more loose stools in 24 hours, abdominal pain, or vomiting with the onset of symptoms between January 16 and February 16, 2023. Active surveillance was conducted by reviewing the affected individuals at the campus and patient registers at the dispensary, where cases were treated. Stool samples, rectal swabs, water samples, and suspected food samples were collected for microbiological testing using conventional culture, multiplex polymerase chain reaction (PCR), and kit-based real-time PCR methods. Results Out of 1,850 residents, 106 (5.7%) were affected by gastrointestinal symptoms like diarrhea, vomiting, etc. The attack rate for females was 23 (1.24%) and for males was 83 (4.49%). The most affected individual median age was 21 years (range: 2-51 years). From the laboratory investigations, most of the cases demonstrated polymicrobial etiologies. Gastroenteritis pathogens like Campylobacter, astrovirus, androtavirusdiarrheagenic Escherichia coli (DEC) (EAEC, EIEC, ETEC, EPEC, and EAEC), Shigella,etc., were detected in the suspected samples. The environmental investigation indicated the presence of rusted and leaky water pipes and sewage pipelines, along with ineffective chlorination of the water plant. Conclusions Based on epidemiological and laboratory investigations, it is conjectured that sewage and fecal contamination of drinking water and poor maintenance of the water distribution system most likely caused the outbreak described in this study. Basic treatment modalities, adequate chlorination, and periodic inspection of the water system were suggested, which controlled the outbreak to a greater extent.

Water scarcity and increasing urbanization are forcing municipalities to consider alternative water sources, such as stormwater, to fill in water supply gaps or address hydromodification of receiving urban streams. Mounting evidence suggests that stormwater is often contaminated with human feces, even in stormwater drainage systems separate from sanitary sewers. Pinpointing sources of human contamination in drainage networks is challenging given the diverse sources of fecal pollution that can impact these systems and the non-specificity of traditional fecal indicator bacteria (FIB) for identifying these host sources. As such, we used a toolbox approach that encompassed microbial source tracking (MST), FIB monitoring, and bacterial pathogen monitoring to investigate microbial contamination of stormwater in an urban municipality. We demonstrate that human sewage frequently contaminated stormwater (in >50% of routine samples), based on the presence of the human fecal marker HF183, and often exceeded microbial water quality criteria. Arcobacter butzleri, a pathogen of emerging concern, was also detected in >50% of routine samples, with 75% of these pathogen-positive samples also being positive for the human fecal marker HF183, suggesting human municipal sewage as the likely source for this pathogen. MST and FIB were used to track human fecal pollution in the drainage network to the most likely point source of contamination, for which a sewage cross-connection was identified and confirmed using tracer dyes. These results point to the ubiquitous presence of human sewage in stormwater and also provide municipalities with the tools to identify sources of anthropogenic contamination in storm drainage networks.IMPORTANCEWater scarcity, increased urbanization, and population growth are driving municipalities worldwide to consider stormwater as an alternative water source in urban environments. However, many studies suggest that stormwater is relatively poor in terms of microbial water quality, is frequently contaminated with human sewage, and therefore could represent a potential health risk depending on the type of exposure (e.g., irrigation of community gardens). Traditional monitoring of water quality based on fecal bacteria does not provide any information about the sources of fecal pollution contaminating stormwater (i.e., animals/human feces). Herein, we present a case study that uses fecal bacterial monitoring, microbial source tracking, and bacterial pathogen analysis to identify a cross-connection that contributed to human fecal intrusion into an urban stormwater network. This microbial toolbox approach can be useful for municipalities in identifying infrastructure problems in stormwater drainage networks to reduce risks associated with water reuse.

Concerns are rising about the contamination of recreational waters from human and animal waste, along with associated risks to public health. However, existing guidelines for managing pathogens in these environments have not yet fully integrated risk-based pathogen-specific criteria, which, along with recent advancements in indicators and markers, are essential to improve the protection of public health. This study aimed to establish risk-based critical concentration benchmarks for significant enteric pathogens, i.e., norovirus, rotavirus, adenovirus, Cryptosporidium spp., Giardia lamblia, Campylobacter jejuni, Salmonella spp., and Escherichia coli O157:H7. Applying a 0.036 risk benchmark to both marine and freshwater environments, the study identified the lowest critical concentrations for children, who are the most susceptible group. Norovirus, C. jejuni, and Cryptosporidium presented lowest median critical concentrations for virus, bacteria, and protozoa, respectively: 0.74 GC, 1.73 CFU, and 0.39 viable oocysts per 100 mL in freshwater for children. These values were then used to determine minimum sample volumes corresponding to different recovery rates for culture method, digital polymerase chain reaction and quantitative PCR methods. The results indicate that for children, norovirus required the largest sample volumes of freshwater and marine water (52.08 to 178.57 L, based on the 5th percentile with a 10 % recovery rate), reflecting its low critical concentration and high potential for causing illness. In contrast, adenovirus and rotavirus required significantly smaller volumes (approximately 0.24 to 1.33 L). C. jejuni and Cryptosporidium, which required the highest sampling volumes for bacteria and protozoa, needed 1.72 to 11.09 L and 4.17 to 25.51 L, respectively. Additionally, the presented risk-based framework could provide a model for establishing pathogen thresholds, potentially guiding the creation of extensive risk-based criteria for various pathogens in recreational waters, thus aiding public health authorities in decision-making, strengthening pathogen monitoring, and improving water quality testing accuracy for enhanced health protection.

Despite the prevalence of co-infections and the association of over 50 viral and 46 bacterial pathogens with pig diseases, little is known about their simultaneous occurrence, particularly in commercial pig farming environments where health programs are in place. To address this knowledge gap, this study aimed to evaluate the pathogen threshold of respiratory and enteric pathogens in pig herds using the Pork MultiPath™ (PMP1 and PMP2, respiratory and enteric respectively) technology, which detects multiple pathogens simultaneously in a single reaction with high sensitivity and specificity. In this study the most prevalent respiratory pathogens, Mycoplasma hyrohinis, Pasteurella multocida, and Haemophilus parasuis detected by PMP1 were effectively controlled during the nursery stage through strategic treatment with tiamulin. Even though the major respiratory incidences were reduced, the recorded coughing and sneezing rates were associated with the levels of H. parasuis and M. hyrohinis, which were set at 1356 and 1275 copies/reaction, respectively. In addition, one of the identified co-infection patterns indicated a strong relationship between the occurrence of H. parasuis and M. hyorhinis at the sample and pen levels, highlighting the high likelihood of detecting these two pathogens together. Testing with enteric panel PMP2 revealed that the most frequently detected virulence factors during the early nursery stage were Escherichia coli genes for toxins - ST1, ST2, and fimbriae - F4 and F18. Moreover, a co-infection with Rotavirus B and C was often observed during the nursery stage, and a strong positive correlation between these two markers has been identified. Additionally, the levels of several markers, namely E. coli F4, F5, F18, LT, ST1, and ST2, have been associated with a higher likelihood of sickness in pig populations. In addition, the onset of Brachyspira pilosicoli during the nursery and grower stages was found to be associated with an increased risk of diarrhoea, with a set threshold at around 500 copies/reaction. Although simultaneous detection of multiple pathogens is not yet widely used in the pig industry, it offers a significant advantage in capturing the diversity and interactions of co-infections. Testing pooled samples with Pork MultiPath™ is cost-effective and practical to regularly monitor the health status of pig populations.

Plastic pollution is increasingly found in agricultural environments, where it contaminates soil and crops. Microbial biofilms rapidly colonise environmental plastics, such as the plastic mulches used in agricultural systems, which provide a unique environment for microbial plastisphere communities. Human pathogens can also persist in the plastisphere, and enter agricultural environments via flooding or irrigation with contaminated water. In this study we examined whether Salmonella Typhimurium and Vibrio cholerae can be transferred from the plastisphere on plastic mulch to the surface of ready-to-eat crop plants, and subsequently persist on the leaf surface. Both S. Typhimurium and V. cholerae were able to persist for 14 days on fragments of plastic mulch adhering to the surface of leaves of both basil and spinach. Importantly, within 24 h both pathogens were capable of dissociating from the surface of the plastic and were transferred onto the surface of both basil and spinach leaves. This poses a further risk to food safety and human health, as even removal of adhering plastics and washing of these ready-to-eat crops would not completely remove these pathogens. As the need for more intensive food production increases, so too does the use of plastic mulches in agronomic systems. Therefore, there is now an urgent need to understand the unquantified co-pollutant pathogen risk of contaminating agricultural and food production systems with plastic pollution.

UNASSIGNED: This study discussed the effect of probiotic supplementation on laying hens\' diets and the enhancement of egg quality during the cold storage period.
UNASSIGNED: To study the efficacy of the addition of probiotics to hen diets in terms of improving the egg\'s quality during the cold storage period and protection against enteric pathogens.
UNASSIGNED: 100 table eggs were collected from farms of laying hens on a battery system, 46 weeks old HylineW36 white in Sharkia Government. The collected eggs were separated into 2 groups (50 each); the control group from hens fed on diets without probiotics, and the probiotic group from hens fed on diets with (100 g/ton) of supplemented probiotics preparation. All groups were separated into 5 sub-groups for the examinations; on the fresh day, 7th, 14th, 21st, and 28th days on cold storage at 4°C. Chemical, physical, and microbiological examinations were done for internal egg contents and eggshells.
UNASSIGNED: Our results showed that probiotics supplements have advantageous effects on the quality of eggs during cold storage periods. Also, microbiological examination proved that eggshells from hens fed on diets with probiotics supplemented (100 g/ton) have decreased the level of bacterial contamination with Salmonella and Escherichia coli than hens fed on a regular diet.
UNASSIGNED: It could be shown that the probiotics supplementation may decrease and reduce the effect of the storage period on the quality of shell, albumen, and yolk.

Glucuronidation is a detoxification process to eliminate endo- and xeno-biotics and neurotransmitters from the host circulation. Glucuronosyltransferase binds these compounds to glucuronic acid (GlcA), deactivating them and allowing their elimination through the gastrointestinal (GI) tract. However, the microbiota produces β-glucuronidases that release GlcA and reactivate these compounds. Enteric pathogens such as enterohemorrhagic Escherichia coli (EHEC) and Citrobacter rodentium sense and utilize galacturonic acid (GalA), an isomer of GlcA, to outcompete the microbiota promoting gut colonization. However, the role of GlcA in pathogen colonization has not been explored. Here, we show that treatment of mice with a microbial β-glucuronidase inhibitor (GUSi) decreased C. rodentium\'s colonization of the GI tract, without modulating bacterial virulence or host inflammation. Metagenomic studies indicated that GUSi did not change the composition of the intestinal microbiota in these animals. GlcA confers an advantage for pathogen expansion through its utilization as a carbon source. Congruently mutants unable to catabolize GlcA depict lower GI colonization compared to wild type and are not sensitive to GUSi. Germfree mice colonized with a commensal E. coli deficient for β-glucuronidase production led to a decrease of C. rodentium tissue colonization, compared to animals monocolonized with an E. coli proficient for production of this enzyme. GlcA is not sensed as a signal and doesn\'t activate virulence expression but is used as a metabolite. Because pathogens can use GlcA to promote their colonization, inhibitors of microbial β-glucuronidases could be a unique therapeutic against enteric infections without disturbing the host or microbiota physiology.