Enteropathogenic Bacteria Research Articles

Antibacterial and antibiofilm activity of human gut lactic acid bacteria.

The present study focused on the antibacterial and antibiofilm activity of novel lactic acid bacterial (LAB) strains isolated from the healthy human volunteers of different age groups and their consortium (LABCON), against the enteropathogenic bacteria. From the study, methanolic extract of LAB isolates and their consortia were found to have promising antibacterial activity and antibiofilm activity against Escherichia coli (ATCC 35218) and Staphylococcus aureus (ATCC 25923). The antimicrobial compounds including the DL-3 phenyllactic acid, DL-p-hydroxyphenyllactic acid, and Succinic acid produced by the LAB could be considered to inhibit the growth and biofilm formation by E. coli (ATCC 35218) and S. aureus (ATCC 25923). Detailed insight into the antibiofilm activity could also be demonstrated by Confocal Raman microscopy attached with AFM and Fluorescent microscope. From the results of the study, the consortium LABCON was superior in antimicrobial and antibiofilm activity and can be considered to have promising application in infection control.

Shigella Senses the Environmental Cue Leucine to Promote its Virulence Gene Expression in the Colon

Shigella is a foodborne enteropathogenic bacteria that causes severe bacillary dysentery in humans. Shigella primarily colonizes the human colon and causes disease via invasion of colon epithelial cells. However, the signal regulatory mechanisms associated with its colonization and pathogenesis in the colon remain poorly defined. Here, we report a leucine-mediated regulatory mechanism that promotes Shigella virulence gene expression and invasion of colon epithelial cells. Shigella in response to leucine, which is highly abundant in the colon, via the leucine-responsive regulator Lrp and the binding of Lrp with leucine induces the expression of a newly identified small RNA SsrV. SsrV then activates the expression of virF and downstream invasion-related virulence genes by increasing the protein level of the LysR-type transcription regulator LrhA, therefore enabling Shigella invasion of colon epithelial cells. Shigella lacking ssrV displays impaired invasion ability. Collectively, these findings suggest that Shigella employs a leucine-responsive environmental activation mechanism to establish colonization and pathogenicity.

A New Real-Time Simple Method to Measure the Endogenous Nitrate Reductase Activity (Nar) in Paracoccus denitrificans and Other Denitrifying Bacteria.

The transmembrane nitrate reductase (Nar) is the first enzyme in the dissimilatory alternate anaerobic nitrate respiratory chain in denitrifying bacteria. To date, there has been no real-time method to determine its specific activity embedded in its native membrane; here, we describe such a new method, which is useful with the inside-out membranes of Paracoccus denitrificans and other denitrifying bacteria. This new method takes advantage of the native coupling of the endogenous NADH dehydrogenase or Complex I with the reduction of nitrate by Nar through the quinone pool of the inner membranes of P. denitrificans. This is achieved under previously reached anaerobic conditions. Inner controls confirming the specific Nar activity determined by this new method were made by the total inhibition of the Nar enzyme by sodium azide and cyanide, well-known Nar inhibitors. The estimation of the Michaelis-Menten affinity of Nar for NO3- using this so-called Nar-JJ assay gave a Km of 70.4 μM, similar to previously determined values. This new Nar-JJ assay is a suitable, low-cost, and reproducible method to determine in real-time the endogenous Nar activity not only in P. denitrificans, but in other denitrifying bacteria such as Brucella canis, and potentially in other entero-pathogenic bacteria.

Paracellular Delivery of Protein Drugs with Smart EnteroPatho Nanoparticles.

A general platform for the safe and effective oral delivery of biologics would revolutionize the administration of protein-based drugs, improving access for patients and lowering the financial burden on the health-care industry. Because of their dimensions and physiochemical properties, nanomaterials stand as promising vehicles for navigating the complex and challenging environment in the gastrointestinal (GI) tract. Recent developments have led to materials that protect protein drugs from degradation and enable controlled release in the small intestine, the site of absorption for most proteins. Yet, once present in the small intestine, the protein must transit through the secreted mucus and epithelial cells of the intestinal mucosa into systemic circulation, a process that remains a bottleneck for nanomaterial-based delivery. One attractive pathway through the intestinal mucosa is the paracellular route, which avoids cell trafficking and other degradative processes in the interior of cells. Direct flux between cells is regulated by epithelial tight junctions (TJs) that seal the paracellular space and prevent protein flux. Here, we describe a smart nanoparticle system that directly and transiently disrupts TJs for improved protein delivery, an unrealized goal to-date. We take inspiration from enteropathogenic bacteria that adhere to intestinal epithelia and secrete inhibitors that block TJ interactions in the local environment. To mimic these natural mechanisms, we engineer nanoparticles (EnteroPatho NPs) that attach to the epithelial glycocalyx and release TJ modulators in response to the intestinal pH. We show that EnteroPatho NPs lead to TJ disruption and paracellular protein delivery, giving rise to a general platform for oral delivery.

Effects of a Nutraceutical Treatment on the Intestinal Microbiota of Sled Dogs.

Dog sledding is the main discipline of working dogs on snow, consisting of a team of dogs pulling a sled under the guidance of the owner. To carry out this sport, dogs must have adequate nutrition and vitamin and antioxidant supplementation to ensure that the physical effort is optimal. The present study evaluated the effect that sporting activity and stress have on the canine intestinal microbiota by dividing the dogs into two groups: a control group that did not take any nutraceutical products and the treated group to which a nutraceutical product was administered. The nutraceutical administered in this study is used in all cases of canine intestinal dysbiosis in which it is essential to quickly restore a balanced intestinal microbiota. The results obtained show that in dogs not taking the nutraceutical, there is an increase in bacteria, such as Streptococcus spp. and E. coli, considered enteropathogenic to the detriment of beneficial bacterial species such as Faecalibacterium spp., Turicibacter spp., Blautia spp., Fusobacterium spp., and Clostridium hiranonis. Instead, the group of dogs treated with nutraceutical displays a lower amount of enteropathogenic bacteria and a great increase in the other bacterial species considered beneficial for the animal's health. The results obtained in the present study show that Microbiotal cane® can be used in dogs subject to intense sporting activity by preventing severe alterations at intestinal ecosystem levels by maintaining intestinal bacterial composition as balanced as possible.

Novel Insights into the Interaction between Enteropathogenic Bacteria, Pyroptosis and IBD.

Inflammatory bowel disease (IBD) is a chronic and recurrent inflammatory disease of the intestinal tract. The complex pathophysiological mechanisms of IBD include genetic susceptibility, environmental factors, and abnormal immune response of the gut microbiota. Gut microbiota forms a metabolic organ that contributes to human health by performing various physiological functions. The development of IBD is closely linked to the imbalance of gut microbiota. In IBD patients, this imbalance is mainly characterized by an increased abundance of pro-inflammatory microorganisms, specifically enteropathogenic bacteria. Pyroptosis is a form of programmed cell death that can be initiated by microbial infection or host factors. It occurs mostly after intracellular infection with bacteria or pathogens. Other than cell death, its primary effect is to release inflammatory mediators that trigger an inflammatory response in the host. Pyroptosis is an important component of innate immunity and can protect against intracellular risk factors via the inflammatory response. However, excessive activation can cause disease. Previous studies of IBD have indicated a complex relationship between gut microbiota and pyroptosis. Some enteropathogenic bacteria can activate the host's immune system to clear infected cells. This inhibits the proliferation of enteropathogenic bacteria by inducing pyroptosis and restoring the balance of gut microbiota. However, the initial inflammatory response and damage to the integrity of the intestinal barrier are crucial factors that elicit the onset of IBD and favor its progression. This review summarizes research on the role of several common enteropathogenic bacteria in the development of IBD through their induction of host cell pyroptosis. A better understanding of the complex interactions between gut microbiota and pyroptosis should lead to the identification of new targets and treatment options for IBD.

MassARRAY: a high-throughput solution for rapid detection of foodborne pathogens in real-world settings.

Bacterial foodborne pathogens pose a substantial global public health concern, prompting government agencies and public health organizations to establish food safety guidelines and regulations aimed at mitigating the risk of foodborne illness. The advent of DNA-based amplification coupled with mass spectrometry, known as MassARRAY analysis, has proven to be a highly precise, sensitive, high-throughput, and cost-effective method for bacterial detection. This study aimed to develop, validate, and evaluate a MassARRAY-based assay for the detection and identification of significant enteropathogenic bacteria. The MassARRAY-based assay was developed for the detection of 10 crucial bacterial foodborne pathogens, including Campylobacter coli, Campylobacter jejuni, Clostridium perfringens, Escherichia coli, Enterococcus faecalis, Enterococcus faecium, Listeria monocytogenes, Salmonella spp., Shigella spp., and Staphylococcus aureus. The assay was optimized using the reference gDNA (n = 19), followed by validation using gDNA (n = 85) of reference and laboratory isolates. Additionally, the evaluation of the assay's reaction using a mixture of gDNA from all nine targeted species was performed. The limit of detection of the developed MassARRAY-based assay was determined using bacterial cells. Moreover, the validation method for field samples was evaluated by comparing it with standard microbiological testing methods routinely analyzed. The developed MassARRAY-based assay demonstrated 100% concordance with known bacterial pure cultures. The assay's reaction using a mixture of gDNA from all nine targeted species revealed the MassARRAY's capability to detect all targeted species in a single assay with the lowest concentration of 1 ng/μL of gDNA. The limits of detection of the assay range from 357 ± 101 to 282,000 ± 79,196 cells. Moreover, the validation of the assay in field samples revealed a 100% correlation between the data obtained from the standard microbiological method and the MassARRAY-based assay. These findings suggested that the developed MassARRAY-based assay exhibited the excellence in high-throughput detection of foodborne bacterial pathogens with high accuracy, reliability, and potential applicability within real-world field samples.

Characterization of a Lactobacillus gasseri strain as a probiotic for female vaginitis

Vaginitis, a prevalent gynecological condition in women, is mainly caused by an imbalance in the vaginal micro-ecology. The two most common types of vaginitis are vaginal bacteriosis and vulvovaginal candidiasis, triggered by the virulent Gardnerella vaginalis and Candida albicans, respectively. In this study, a strain capable of inhibiting G. vaginalis and C. albicans was screened from vaginal secretions and identified as Lactobacillus gasseri based on 16S rRNA sequences. The strain, named L. gasseri VHProbi E09, could inhibit the growth of G. vaginalis and C. albicans under co-culture conditions by 99.07% ± 0.26% and 99.95% ± 0.01%, respectively. In addition, it could significantly inhibit the adhesion of these pathogens to vaginal epithelial cells. The strain further showed the ability to inhibit the enteropathogenic bacteria Escherichia coli and Salmonella enteritidis, to tolerate artificial gastric and intestinal fluids and to adhere to intestinal Caco-2 cells. These results suggest that L. gasseri VHProbi E09 holds promise for clinical trials and animal studies whether administered orally or directly into the vagina. Whole-genome analysis also revealed a genome consisting of 1752 genes for L. gasseri VHProbi E09, with subsequent analyses identifying seven genes related to adhesion and three genes related to bacteriocins. These adhesion- and bacteriocin-related genes provide a theoretical basis for understanding the mechanism of bacterial inhibition of the strain. The research conducted in this study suggests that L. gasseri VHProbi E09 may be considered as a potential probiotic, and further research can delve deeper into its efficacy as an agent which can restore a healthy vaginal ecosystem.

Impact of COVID-19 Restrictions on Incidence of Enteropathogenic Bacteria, Virus, and Parasites in Denmark: A National, Register-Based Study.

Diarrheal diseases caused by enteric pathogens are a significant public health concern. It is widely considered that close contact between persons, poor hygiene, and consumption of contaminated food are the primary causes of gastroenteritis. Clinical microbiology laboratory observations indicate that the incidence of enteropathogenic microorganisms may have been reduced in Denmark during the COVID-19 pandemic. All Departments of Clinical Microbiology in Denmark provided data on the monthly incidence of Salmonella spp., Escherichia coli, Campylobacter spp., Clostridioides difficile, Norovirus GI+GII, Giardia duodenalis, and Cryptosporidium from March 2018 to February 2021. The data were divided into three periods as follows: Control Period 1 (March 2018 to February 2019); Control Period 2 (March 2019 to February 2020); and the Restriction (pandemic) Period (March 2020 to February 2021). The incidences of pathogenic Salmonella spp.-, Escherichia coli-, and Campylobacter spp.-positive samples decreased by 57.3%, 48.1%, and 32.9%, respectively, during the restriction period. No decrease in C. difficile was observed. Norovirus GI+GII-positive samples decreased by 85.6%. Giardia duodenalis-positive samples decreased by 66.2%. Cryptosporidium species decreased by 59.6%. This study demonstrates a clear decrease in the incidence of enteropathogenic bacteria (except for C. difficile), viruses, and parasites during the SARS-CoV-2 restriction period in Denmark.

Monitoring of enteropathogenic Gram-negative bacteria in wastewater treatment plants: a multimethod approach

The wastewater treatment processes are associated with the emission of microbial aerosols, including enteropathogenic bacteria. Their presence in this work environment poses a real threat to the health of employees, both through the possibility of direct inhalation of the contaminated air and indirectly through the pollution of all types of surfaces with such bioaerosol particles. This study aimed to investigate the prevalence of enteropathogenic bacteria in the air, on surfaces, and in wastewater samples collected in four wastewater treatment plants (WWTPs). The effectiveness of conventional culture-biochemical, as well as spectrometric and molecular methods for the rapid detection of enteropathogenic bacteria at workstations related to particular stages of wastewater processing, was also evaluated. Bioaerosol, surface swab, and influent and effluent samples were collected from wastewater plants employing mechanical–biological treatment technologies. The air samples were collected using MAS-100 NT impactor placed at a height of 1.5 m above the floor or ground, simulating aspiration from the human breathing zone. Surface samples were collected with sterile swabs from different surfaces (valves, handles, handrails, and coveyor belts) at workplaces. The raw influent and treated effluent wastewater samples were aseptically collected using sterile bottles. The identification of bacterial entheropathogens was simultaneously conducted using a culture-based method supplemented with biochemical (API) tests, mass-spectrometry (MALDI TOF MS), and molecular (multiplex real-time PCR) methods. This study confirmed the common presence of bacterial pathogens (including enteropathogenic and enterotoxigenic Escherichia coli, Salmonella spp., Campylobacter spp., and Yersinia enterocolitica) in all air, surface, and wastewater samples at studied workplaces. Higher concentrations of enteropathogenic bacteria were observed in the air and on surfaces at workplaces where treatment processes were not hermetized. The results of this study underline that identification of enteropathogenic bacteria in WWTPs is of great importance for the correct risk assessment at workplaces. From the analytical point of view, the control of enteropathogenic bacterial air and surface pollution using rapid multiplex-PCR method should be routinely performed as a part of hygienic quality assessment in WWTPs.

Antibiotic resistance of enteropathogenic bacteria in a teaching hospital in North Khuzestan during a three-year period.

Gastrointestinal infections affect many people annually. The most common bacterial agents involved in these infections are enteropathogenic bacteria and in the continuation of using broad-spectrum antibiotics, Clostridium difficile-associated diarrhea is involved, especially in hospitalized patients. The aim of the present study was to investigate the pattern of antibiotic resistance among enteropathogenic bacteria. In this cross-sectional study, 163 samples of patients with diarrhea in Dezful Ganjavian Hospital were examined. The samples were cultured in MacConkey, Hektoen enteric agar and GN broth, and cycloserine cefoxitin fructose agar media and incubated under standard conditions. In order to identify enteropathogenic bacteria, biochemical tests and serological confirmatory tests were used. Antibiotic resistance pattern of the isolates was investigated by Kirby-Bauer disk diffusion susceptibility test. The frequency of pathogenic bacteria includes 41.1% of Shigella flexneri, followed by 41.1% of S. sonnei, 6.7% of Enteropathogenic E. coli, 5.5% of Salmonella enterica Serogroup B, and 5.5% of Shigella dysenteriae. The results revealed a total of 46 patients with orders regarding C. difficile culture, no C. difficile was isolated from the samples. The studied isolates showed the highest resistance to trimethoprim-sulfamethoxazole, and ceftriaxone (88.3%), and the most effective antibiotic in the treatment of patients was ciprofloxacin with 86% sensitivity. Susceptibility to antibiotics was different among the isolates, which shows that the early identification of the infection agent and the selection of the correct antibiotic treatment are effective in improving the gastrointestinal infection and preventing the spread of the infection.

Theophylline-based control of repA on a Clostridioides difficile plasmid for use in allelic exchange

Historically, mutagenesis in the non-model enteropathogenic bacterium Clostridioides difficile has been challenging. Developing a versatile and reliable method of generating targeted mutations in C. difficile is important to further our understanding of its pathogenesis. Some of the most common targeted mutagenesis systems rely on allelic exchange mediated by either uracil auxotrophy combined with a toxic uracil precursor, a toxin/anti-toxin system, group II introns, or CRISPR/Cas mutagenesis. However, each of these methods suffers from its own issues. Here, we develop and test an allelic exchange strategy which better facilitates screening for integration and selecting for excision than previous systems. This is achieved by controlling plasmid replication with a theophylline-dependent riboswitch cloned upstream of repA, the gene whose product controls plasmid replication. This allows efficient mutant generation, can be performed in a wild-type strain of C. difficile, does not have the off-target effects inherent to group II introns, and alleviates the problem of testing multiple gRNA targets in CRISPR mutagenesis.

Interaction between Dietary Lactoferrin and Gut Microbiota in Host Health.

The gut microbiota are known to play an important role in host health and disease. Alterations in the gut microbiota composition can disrupt the stability of the gut ecosystem, which may result in noncommunicable chronic diseases (NCCDs). Remodeling the gut microbiota through personalized nutrition is a novel therapeutic avenue for both disease control and prevention. However, whether there are commonly used gut microbiota-targeted diets and how gut microbiota-diet interactions combat NCCDs and improve health remain questions to be addressed. Lactoferrin (LF), which is broadly used in dietary supplements, acts not only as an antimicrobial in the defense against enteropathogenic bacteria but also as a prebiotic to propagate certain probiotics. Thus, LF-induced gut microbiota alterations can be harnessed to induce changes in host physiology, and the underpinnings of their relationships and mechanisms are beginning to unravel in studies involving humans and animal models.

Type 1 fimbriae-mediated collective protection against type 6 secretion system attacks.

Bacterial competition may rely on secretion systems such as the type 6 secretion system (T6SS), which punctures and releases toxic molecules into neighboring cells. To subsist, bacterial targets must counteract the threats posed by T6SS-positive competitors. In this study, we used a comprehensive genome-wide high-throughput screening approach to investigate the dynamics of interbacterial competition. Our primary goal was to identify deletion mutants within the well-characterized E. coli K-12 single-gene deletion library, the Keio collection, that demonstrated resistance to T6SS-mediated killing by the enteropathogenic bacterium Cronobacter malonaticus. We identified 49 potential mutants conferring resistance to T6SS and focused our interest on a deletion mutant (∆fimE) exhibiting enhanced expression of type 1 fimbriae. We demonstrated that the presence of type 1 fimbriae leads to the formation of microcolonies and thus protects against T6SS-mediated assaults. Collectively, our study demonstrated that adhesive structures such as type 1 fimbriae confer collective protective behavior against T6SS attacks.IMPORTANCEType 6 secretion systems (T6SS) are molecular weapons employed by gram-negative bacteria to eliminate neighboring microbes. T6SS plays a pivotal role as a virulence factor, enabling pathogenic gram-negative bacteria to compete with the established communities to colonize hosts and induce infections. Gaining a deeper understanding of bacterial interactions will allow the development of strategies to control the action of systems such as the T6SS that can manipulate bacterial communities. In this context, we demonstrate that bacteria targeted by T6SS attacks from the enteric pathogen Cronobacter malonaticus, which poses a significant threat to infants, can develop a collective protective mechanism centered on the production of type I fimbriae. These adhesive structures promote the aggregation of bacterial preys and the formation of microcolonies, which protect the cells from T6SS attacks.

Understanding the Risk Landscape: Analyzing Factors Impacting Food Vending in Nigeria

Even in the modern era with increased awareness about foodborne illness, it still remains a serious public health issue, everywhere around the world, and food handling, preparation and service are crucial practices for food safety. The factor of cookware preparation, storage systems, and/or the versatility and level of education of food handlers are all types of food safety factors that directly or indirectly influence food safety to some degree. Properly nailing down food hygiene first entails looking closely into personal hygiene and the conduct of employees working in food businesses. The food handlers with poor personal hygiene may, in close proximity to the food, be carriers of intestinal helminths, protozoa, and enteropathogenic bacteria. This is when the hands of the handlers can carry excreta from their feces, and they spread it into food with their fingers, thereby leading to infections in the customers. This study assessed the most influential risk factors causing food vendor's illness, identifies several of them, like the groceries, raw food and quality; food prepared and handled in the streets; transportation; absence of regulations involving time and specific holding temperature; and the environment's cleanliness. Nevertheless, the research work came to reveal that various aspects of street food safety are persistently being taken into much account in consuming what belongs to dissimilar socio-economic grouping. Therefore, since the economic benefits of street food trading are incalculable, it is very critical that authorities strengthened and implemented the policies/ regulations in order to ensure food safety. Keywords: Foodborne illnesses, Food safety, Food handling, Food vendors, Street food

Reduction of Bacterial Load on Broiler Carcasses Using Low-Volume Fluidic Nozzles in Combination with 60 °C Water at 450 Psi Pressure

With the changing regulations in poultry processing, increased pressure is placed on integrators to reduce the number of human enteropathogenic bacteria on the final carcass and/or parts. Reducing the total number of bacteria on broiler carcasses before entering the evisceration side of the processing plant is projected to reduce the number of bacteria on the carcasses after chilling. This study was designed to evaluate the efficacy of a prototype wash cabinet using low volume, fluidic nozzles in combination with high pressure (450 psi) and hot water (60 °C) to remove bacteria from pre-scald, post-scald, or post picked carcasses. Carcasses (n = 5) from each location were obtained from a commercial processing plant, placed into individual sterile sample bags, placed into an insulated container, and transported to the U.S. National Poultry Research Center Pilot Plant within 30 min of collection. Carcasses were hung in standard shackles and sampled pre-wash with pre-moistened, cellulose swabs. All carcasses were washed in the prototype wash cabinet with 60 °C water at 450 psi at a line speed of 52 birds/minute on 15.24 cm centered shackles. Post-wash breast sponge samples were collected identical to pre-wash swabs. Buffered peptone water (BPW) was added, sponges stomached and serially diluted before plating onto total aerobic count (TAC), Enterobacteriaceae (ENT) and Escherichia. coli (EC) Petrifilm® cards. All PetriFilm® cards were incubated at 37 °C for 24 ± 2 h. After incubation, bacterial counts were recorded and converted to log10 CFU/swab. Samples were processed for Campylobacter species using the Tempo® CAM protocol. Four replications were conducted on separate dates. Paired t-tests were used to compare numbers recovered from breast swabs collected before and after the wash cabinet, significance reported at p < 0.05. Pre-scald samples had significant reductions of 2.50, 2.01, and 1.73 log10 colony-forming units/carcass (CFU/carcass) for TAC, Ent, and EC Petrifilm®, respectively, and a 2.21 CFU/mL reduction of Campylobacter species using Tempo® CAM. Post-scald, there were significant reductions of 2.09, 1.23, and 0.90 CFU/carcass for TA, Ent, and EC Petrifilm®, respectively, and a 1.14 CFU/mL reduction of Campylobacter species using Tempo® CAM. Post-pick, significant reductions of 0.73, 1.53, and 0.99 CFU/carcass for TA, Ent, and EC Petrifilm®, respectively, and a 0.86 CFU/carcass reduction of Campylobacter species using Tempo® CAM were reported. These data indicate that hot water at high pressure can reduce total bacterial load on carcasses and reduce pathogenic bacteria on carcasses prior to evisceration.

Antibiotic Susceptibility of Lactic Acid Bacteria Isolated From Cow, Goat, Donkey, Buffalo, Sheep, Camel and Human Milk

Breast milk is a very important factor in regulating gastrointestinal function, improving the immune system and preventing acute illnesses (e.g. acute otitis media), especially during breastfeeding. Breast milk allows beneficial bacteria such as bifidobacteria and lactobacillus species to colonise the newborn intestin*e instead of potential enteropathogenic bacteria such as streptococci and escherichia coli. The aim of the study was to compare the antibiotic resistance of Lactic acid bacteria members isolated from different milk samples. Six milk samples were collected from each of seven different milk sources (donkey, goat, cow, buffalo, sheep, camel and human). The MRS and M-17 medium were used with the double layer sandwich method for isolation of LAB members selected from typic colonies, gram-positive, catalase-negative used in the study. The isolated 42 LAB species were determined antibiotic susceptibility with 9 standart antibiotic discs such as tetracycline, penicillin, kanamycin, streptomycin, rifampin, gentamicin, chloramphenicol, teicoplanin, ciprofloxacin by agar disc diffusion assay test. The 18 LAB isolate were resistant to penicillin, 16 isolate to kanamycin, 14 isolate to gentamicin. LAB isolates such as H1, H4, H5 from Human milk and LAB isolate such as D1 from donkey milk was considered as MDR isolates because they were resistant to at least four of the tested drugs. Among the milks, sheep, camel, buffalo and goat milks were found to be more sensitive to antibiotics on average in the group.

Антибиотикорезистентность Enterobacteriaceae в микробиомах цыплят-бройлеров

Antibiotics have long been overused for non-therapeutic purposes. As a result, Escherichia, Enterobacter, Klebsiella, and Proteus in avian microbiomes have become reservoirs for genetic determinants of resistance, thus spreading resistance to antibiotics and contaminating raw materials and finished products. The food industry is looking for alternative means to preserve health and maintain high productivity of commercial poultry, e.g., probiotics, phytobiotics, organic acids, etc. The research featured the effect of antibiotics and phytobiotics on enteropathogenic bacteria in the microbiomes of broiler chicken.
 Escherichia coli bacteria were cultivated in vitro in subthreshold concentrations on nutrient media with antibiotics for 37 days to study the effect of low doses of antibiotics on the sensitivity of isolates. The study involved microbiocenoses of broiler chicken that received avilamycin A and a phytobiotic based on Brassica juncea, Linum usitatissimum, and Nigella sativa L. A set of experiments covered the species composition of opportunistic Enterobacteriaceae, the phenotypic sensitivity to antibiotics, and the genetic determinants of resistance, as well as the antimicrobial potential of phytobiotics.
 E. coli developed no resistance for 37 days when the antibiotic dose remained below minimal inhibitory. Opportunistic gram-negative Enterobacterales predominated in all litter samples. E. coli, Klebsiella pneumoniae, and Proteus mirabilis accounted for more than 30% of all isolated strains. Avilamycin A and the phytobiotic affected the coccal microflora but had no effect on the genus-species composition of Enterobacteriaceae. Litter samples from both experimental and control groups demonstrated K. pneumoniae with severe phenotypic resistance to ciprofloxacin, as well as blaDHA genes. In broiler farming, maintenance and circulation of resistance agents depends on litter microbiota. In this research, the chicken that received the phytobiotic showed the lowest level of resistance to ciprofloxacin while the groups that received avilamycin A had the highest resistance results.
 During a broiler’s life span, bacteria with no active resistance determinants in their genome remained sensitive to antibiotics, even though the contact with the latter was constant. Phytobiotics showed good prospects for broiler farming as food additive that could reduce and eventually eliminate the intake of antibiotics.

Crohn's disease-associated Escherichia coli LF82 in the gut damage of germ-free honeybees: A laboratory study

Escherichia coli LF82 (LF82) is associated with Crohn's disease. The simplicity and genetic maneuverability of honeybees' gut microbiota make them suitable for studying host-microbe interactions. To understand the interaction between LF82 and host gut, LF82 was used to infect germ-free honeybees (Apis mellifera) orally. We found that LF82 successfully colonized the gut and shortened the lifespan of germ-free bees. LF82 altered the gut structure and significantly increased gut permeability. RT–qPCR showed that LF82 infection activated anti-infective immune pathways and upregulated the mRNAs levels of antimicrobial peptides in the gut of germ-free bees. The gut transcriptome showed that LF82 significantly upregulated genes involved in Notch signaling, adhesion junctions, and Toll and Imd signaling pathways and downregulated genes involved in the peroxisome proliferator-activated receptor (PPAR) signaling pathway, protein digestion and absorption, and tyrosine metabolism. In conclusion, the human-derived enteropathogenic bacterium LF82 can successfully colonize the gut of germ-free honeybees and cause enteritis-like changes, which provides an ideal model organism for revealing the pathogenesis of bacterial-associated diseases.

Isocaloric diets with varying protein levels affected energy metabolism in young adult Sprague-Dawley rats via modifying the gut microbes: A lipid imbalance was brought on by a diet with a particularly high protein content

Protein is the most important macro-nutrient when it comes to maximizing health, body composition, muscle growth, and recovery of body tissue. In recent years, it has been found that protein also plays an important role in metabolism and gut microbiota. This study was performed to investigate the effects of an isocaloric diet with different crude protein contents on the energy metabolism of Sprague-Dawley (SD) rats. Results revealed that compared with the 20% crude protein (CP; control) diet, the 38% CP diet improved serum parameters that are associated with dyslipidemia and glucose metabolic disorders in SD rats, whereas the 50% CP diet increased liver injury indicators and fatty acid synthesis-related genes and protein expression in the liver. Compared with the control diet, the 14% CP diet increased the abundance of colonic short-chain fatty acid-producing bacteria (Lachnospiraceae_NK4A136_group and Ruminiclostridium_9) and promoted colonic microbial cysteine and methionine metabolism, the 38% CP diet up-regulated colonic microbial lysine biosynthesis and degradation pathways, and the 50% CP diet down-regulated colonic mucosal cholesterol metabolism. Furthermore, the increase of multiple colonic enteropathogenic bacteria in the 50% CP group was associated with higher palmitic acid and stearic acid concentrations in the colonic microbes and lower cholesterol and arachidonic acid concentrations in the colonic mucosa. These findings revealed that the 14% CP and 38% CP diets improved rats' energy metabolism, while the 50% CP diet was accompanied by lipid metabolism imbalances and an increase in the abundance of multiple enteropathogenic bacteria.