We report shotgun metagenomic sequence data from milk and fecal samples of dairy cattle in Thailand. This data set captures microbial genetic profiles from mammary- and gut-associated sample types and provides a resource for future comparative microbiome, functional, and antimicrobial resistance gene analyses in dairy cattle.

Cardiac mitochondrial dysfunction is a crucial mechanism underlying obesity-induced cardiovascular diseases. A close link between obesity and gut microbiota has been revealed, and the benefits of gut microbiota modulation by probiotics have been widely identified. Although the probiotic Lactobacillus reuteri KUB-AC5 exerted anti-inflammatory activity and enhanced the activity of beneficial microbes, the effects of this probiotic on mitochondria in the obese heart have never been investigated. Male Wistar rats were divided into four groups to receive either a normal diet (ND; n = 9) or a high-fat and high-calorie diet (HFCD; n = 30) for 24 weeks. At the beginning of week 13, ND-fed rats received vehicle, while HFCD-fed rats were further subdivided into three groups (n = 10/group) to receive either vehicle, a live probiotic Lactobacillus reuteri KUB-AC5, or a heat-killed probiotic Lactobacillus reuteri KUB-AC5. At the end of week 24, cardiac functions were evaluated. The rats were then euthanized to enable blood and cardiac ventricle collection. Evidence from the obese rats treated with Lactobacillus reuteri KUB-AC5 in both forms indicated reduced body weight and attenuated insulin resistance. Regarding the heart, obese rats treated with either form of Lactobacillus reuteri KUB-AC5 had improved cardiac functions, mitochondrial dynamics, mitochondrial biogenesis, fat and ketone body utilization, anaplerosis, ATP production, and oxidative phosphorylation. In addition, treatment with this probiotic diminished oxidative stress and restored the antioxidative capacity of cardiac mitochondria. Our preclinical findings in male rats highlighted the benefits of the probiotic Lactobacillus reuteri KUB-AC5, given in both live and heat-killed forms, in alleviating obesity-induced cardiac mitochondrial dysfunction.

Background: The emergence of multidrug-resistant (MDR) uropathogenic Escherichia coli (UPEC) poses a significant public health challenge, and alternative treatments are urgently needed. Methods: Here, we identified clinical MDR-UPEC strains AT82 and AT84 collected from hospitalized patients that display extensive antimicrobial resistance at both genetic and phenotypic levels. Due to their high resistance profile, we systematically customized a phage cocktail from our coliphage library using hierarchical clustering based on host specificity and candidate selection through bacterial suppression profiles. Results: This pipeline yielded four lytic coliphages, designated Phi25-4, Phi25-6, Phi50-4, and Killian. Their genomes are relatively large ranging from 112–169 kbp and cluster into two distinct lineages comprising two closely related groups: Phi25-4/Phi50-4 and Phi25-6/Killian. Although each phage exhibited potent antibacterial activity, none alone sustained bacterial suppression during prolonged treatment. To overcome this limitation, we systematically compared the antibacterial activity of all possible phage combinations. Conclusions: The four-phage cocktail outperformed all two- or three-phage formulations, sustaining significant growth inhibition of AT82 and AT84 for up to 16 h and reducing area under the curve by more than 80% relative to controls. Cocktail potency was dose-dependent, with lower phage doses yielding the least viable cells at 48 h. Additionally, this cocktail exerted prophylactic action, significantly reducing UPEC invasion by several orders of magnitude, while the phage cocktail alone induced minimal proinflammatory cytokine responses in human bladder epithelium. Together, these findings provide an effective phage cocktail and a complementary framework for cocktail design against urinary tract infections caused by MDR bacteria.

Acute non-typhoidal salmonellosis (NTS) from non-typhoidal Salmonella remains a major cause of foodborne bacterial gastroenteritis, and non-antibiotic interventions are needed to combat multidrug-resistant NTS. Bioactive compounds from edible mushroom extracts have shown both direct and indirect antimicrobial activities on Salmonella. However, the variation in their antimicrobial activity could be due to several factors, including the extract's form and strain. This study investigated the ability of crude exopolysaccharides (EPs) produced by Schizophyllum commune CMU-01 to limit Salmonella infection in vitro. Agar well diffusion and liquid culture were used to determine the direct anti-Salmonella activity of S. commune EPs, while the gentamicin protection assay and qPCR in human gut epithelium (T84 cells) and murine macrophages (RAW264.7 cells) were used to investigate its indirect (immunomodulatory) activity. Our data reveal that S. commune EPs do not confer the direct antimicrobial property against Salmonella. However, its immunomodulatory activity in two important components of the gut innate defense (the gut epithelium and macrophages) against Salmonella infection has been demonstrated. S. commune EPs reduce Salmonella gut epithelial cell invasion and activate macrophages toward M1 (inflammatory phenotype) polarization, resulting in the reduction in intracellular Salmonella burdens. Alterations in proinflammatory and anti-inflammatory cytokine gene expressions were also detected in S. commune EPs-treated cells. These findings suggest that the host innate immune response to fungal exopolysaccharides derived from S. commune CMU-01 favors reducing Salmonella proliferation within host cells by altering the expression levels of proinflammatory cytokines.

Urinary tract infections (UTIs) caused by uropathogenic Escherichia coli (UPEC) are one of the most common bacterial infections in humans. The rise of multidrug-resistant UPEC strains increases the urgent need for alternative treatment. Two diverse lytic bacteriophages (phages), SR02 and SR04, recently exhibited an in vitro anti-UPEC activity. In this study, we reported the interplay among UPEC, phages, and the microenvironment of mammalian urinary tract in UTI phage therapy using both in vitro (human bladder cell line) and in vivo (murine acute UTI) models. A gentamicin protection invasion assay was performed in UPEC-infected human bladder cells (UM-UC-3). Both monophages and the phage cocktail significantly reduced UPEC invasion into UM-UC-3 with a synergistic effect between SR02 and SR04. Female C57BL/6 mice were transurethrally infected with 107 colony-forming units of UPEC, and 2 h later, 108 plaque-forming units of monophages and cocktail were single transurethrally administered to the mouse bladder. At 24 h post-UPEC infection, the cocktail significantly reduced UPEC colonization in the mouse bladder and kidney, but not in the urine. The synergism between SR02 and SR04 was observed only in the mouse bladder. Both monophages and cocktail markedly reduced UPEC ascension into mouse kidneys without a synergism or robust tissue proinflammatory cytokine gene expression. However, increased polymorphonuclear cell infiltration was observed in the bladders of SR04-treated mice. In conclusion, we report the contribution of different host urinary tract microenvironments (urine, bladder, and kidney) in the outcomes of UTI phage therapy with two lytic phages and their combination.

Phage infection undergoes a series of physiological transitions, holding crucial information about phage replication dynamics and potential phage-derived antimicrobials. Although phage-induced cytological changes have been used to infer phage-hijacking mechanisms, current approaches are limited by the lack of comprehensive single-cell morphological analysis and insufficient resolution of temporal dynamics, particularly for phages displaying short latent periods, thereby hindering systematic characterization of morphological transitions throughout the infection cycle. Here, we characterized a newly identified coliphage with a genome of 53 kbp, Tiny, which exhibits an unusually long latent period, making it an ideal candidate for resolving temporal morphological transitions. Tiny exhibits both temperature- and host-dependent killing profiles against diverse Escherichia coli strains, including ATCC 25922, uropathogenic E. coli, and avian pathogenic E. coli. While its replication efficiency is temperature-dependent, showing enhanced productivity at lower temperatures, the duration of its adsorption and latent period is largely host-dependent. Depending on the bacterial strain, Tiny exhibits either a prolonged latent period in slow-adsorbing strains or a rapid one in fast-adsorbing strains, regardless of infection temperature, suggesting phage-host compatibility. Single-cell bacterial cytological profiling of Tiny-infected ATCC 25922 cells revealed that Tiny progressively induces distinct bacterial morphological transitions throughout its lytic cycle, suggesting sequential interference with host physiology as part of its replication cycle prior to cell lysis. This work establishes a broadly applicable framework for dissecting lytic phage biology with high temporal resolution and lays the foundation for future integrative omics studies aimed at understanding how phages sequentially modulate their bacterial hosts.IMPORTANCEAntibiotics trigger unique patterns of morphological changes in bacteria, and these compound-specific signatures provide a basis for determining mechanisms of action in antibiotic discovery. By the same concept, phage-induced morphological changes can reveal key insights into phage replication dynamics and guide the identification of phage-derived antimicrobials. However, the complexity of phage biology and the variability of phage-host interactions pose challenges in interpreting these phenotypic outcomes. Here, we employed a phage-host pair that exhibits an unusually prolonged latent duration as a model to establish a broadly applicable framework for dissecting lytic phage biology with high temporal resolution. Through single-cell bacterial morphological analyses, this approach captures dynamic infection processes inducing morphological transitions across the phage replication cycle. This work provides a phenotypic analysis pipeline to advance our understanding of phage-host interactions and lays the foundation for future integrative omics studies to elucidate how phages sequentially modulate their bacterial hosts.

Acute gastrointestinal (GI) toxicities are common side effects of pelvic radiotherapy in gynecologic cancers. Although bacterial probiotics show promise, there is limited evidence for yeast-based probiotics such as Saccharomyces boulardii. This study aimed to evaluate the efficacy of S. boulardii in preventing acute radiation-induced GI toxicities in patients undergoing pelvic radiotherapy for cervical and endometrial cancers. A prospective, randomized, double-blind, controlled trial was conducted with 60 patients receiving definitive or postoperative pelvic radiotherapy. Patients were randomized to receive either S. boulardii (250 mg/day) or a placebo, starting one week before radiotherapy and continuing throughout treatment. Acute GI toxicities were assessed weekly using the Common Terminology Criteria for Adverse Events (CTCAE) version 5. Stool consistency and inflammatory markers were also monitored. The primary outcome was the reduction in the severity of diarrhea and other GI symptoms. S. boulardii significantly reduced the incidence of type 6 and 7 stool consistency at week 3 (p = 0.01). However, despite this transient improvement, the treatment group experienced a significantly higher incidence of diarrhea at week 4 compared to the control group (p = 0.03). Overall, no significant differences were found between the groups in terms of cumulative GI toxicities or anti-diarrheal medication use, with diarrhea remaining the most common acute GI toxicity in both groups. No consistent effect of S. boulardii was observed across the measured outcomes, and its overall efficacy in preventing acute radiation-induced enteritis remains inconclusive. Further studies with larger sample sizes are warranted. Trial registration: The study was registered with the Thai Clinical Trials Registry (TCTR20210204013) on 04 February 2021.

The discovering new fungal strains, optimal production, and understanding the fundamental aspects of exopolysaccharides (EPs) are important to utilize them in an industrial, medical, and biotechnological perspective. In this study, the optimal conditions for EP production from seven basidiomycetous fungal strains were investigated. The results indicated that six fungal species, Schizophyllum commune, Ganoderma fornicatum, G. williamsianum, Earliella scabrosa, Favolus tenuiculus, and Pycnoporus sanguineus, produced the highest EP yield in potato dextrose broth. The highest yield of EPs produced by Lentinus sajor-caju was obtained in mushroom complete medium broth. It was found that a pH value between 6 and 8 in the liquid culture media promoted EP production. The highest EP yield was obtained for 10 to 14 days which depends on fungal strain. Interestingly, this present study revealed the first report of EP production from G. fornicatum, G. williamsianum, E. scabrosa, F. tenuiculus, and P. sanguineus, including the genera Earliella and Favolus. The obtained crude EPs showed water solubilization ability. The Fourier-transform infrared spectroscopy spectra exhibited typical carbohydrate patterns in all crude EPs. Monosaccharide composition analysis revealed that the crude EPs were primarily composed of glucose, followed by fructose, allose, and allulose, with variations depending on the fungal strain. Additionally, crude EPs demonstrated positive antioxidant potential. Finally, we determined the anti-Salmonella and immunomodulatory effects of crude EPs from S. commune, G. fornicatum, and L. sajor-caju due to their high EP yield. Pretreatment of mouse macrophages with these fungal EPs enhanced the phagocytic killing activity of Salmonella-infected macrophages. Upregulations of pro-inflammatory cytokine expression in macrophages were detected in the fungal EPs-treated groups. Our study reported the optimizing conditions for EP production from several strains of Basidiomycetous fungi and their potential as an alternative to antibiotics for multidrug-resistant Salmonella infection.

We report the draft genome sequences of two extended-spectrum β-lactamase-producing uropathogenic Escherichia coli strains, AT82 and AT84, isolated from patients with urinary tract infections in Thailand. The draft genome sizes are approximately 5,168 kb and 5,164 kb, respectively.

Non-typhoidal salmonellosis (NTS) caused by multidrug-resistant (MDR) Salmonella enterica is a significant public health concern worldwide. Probiotics offer a potential alternative to antibiotics in many infectious diseases, including NTS. However, using living bacteria raises safety concerns in clinical settings, especially in the immunocompromised host. This study compared the anti-Salmonella and immunomodulatory effects between viable (probiotics) and heat-killed (paraprobiotics) lactic acid bacteria Lactiplantibacillus plantarum KUNN19-2 (KUNN19-2), isolated from Thai-style fermented pork (Nham), against several strains of MDR Salmonella. Only viable KUNN19-2 and its cell-free supernatant directly inhibited Salmonella growth by spot-on lawn and agar well diffusion assays. A significant reduction in Salmonella numbers in the co-culture assay with viable KUNN19-2 was observed at 12-14 h after the incubation. Viable and heat-killed KUNN19-2 exhibited moderate adhesion to human colonic epithelium (T84) cells. Pretreatment with either form of KUNN19-2 enhanced macrophage (RAW264.7) phagocytic activity against Salmonella and upregulated pro-inflammatory genes (Mip-2 and Nos2) and anti-inflammatory gene (IL10) expression, with viable KUNN19-2 showing a more potent effect. Collectively, viable KUNN19-2 can directly inhibit Salmonella growth. However, viable and heat-killed KUNN19-2 can modulate gut immunity against Salmonella infection, suggesting that paraprobiotic KUNN19-2 may serve as an alternative treatment against MDR Salmonella through host immune modulation.

Piscine intestinal coccidiosis has been associated with increased susceptibility to secondary bacterial infections, potentially through disruption of the gut microbiota. While many gut bacteria are commensal, infection-related alterations in microbial composition may favor the proliferation of opportunistic taxa. However, the specific effects of intestinal coccidial infections on the gut microbiome of fish remain insufficiently described. This study examined the influence of intestinal coccidial infection on gut microbiota composition and the relative abundance of potentially pathogenic bacteria in juvenile Asian seabass (Lates calcarifer). Intestinal samples from seven coccidia-infected and five uninfected fish were analyzed using 16S rRNA gene sequencing. Infected fish demonstrated greater microbial diversity and marked compositional shifts across multiple taxonomic levels. Relative abundances of genera with reported pathogenic potential, such as Clostridium, Lactococcus, and Kurthia, as well as zoonotic bacteria including Helicobacter and Escherichia-Shigella, were elevated in the infected group. Conversely, uninfected fish harbored higher levels of genera such as Bacillus, Vibrio, Acinetobacter, and Streptococcus, which are commonly associated with environmental niches and opportunistic colonization. Coccidial infection was also linked to microbial imbalance, reflected in reduced Proteobacteria-to-Bacteroidota (P:B) and Proteobacteria-to-Firmicutes (P:F) ratios, alongside histopathological evidence of intestinal inflammation. Intestinal coccidiosis in juvenile Asian seabass is associated with open niche gut characterized by altered microbial diversity and increased abundance of bacterial genera with pathogenic potential. These findings suggest that coccidial infection may contribute to an elevated risk of secondary infections, underscoring the relevance of gut microbiome monitoring in fish health management. Further studies are warranted to clarify host–parasite–microbiome interactions and their implications for aquaculture health.

Urinary tract infections are commonly caused by uropathogenic Escherichia coli (UPEC). Due to the emergence of multidrug-resistant UPEC, rendering antibiotic treatment ineffective, phage combination-based therapy has been proposed as a potential alternative. Here, we present a formulation of a genetically diverse phage-derived cocktail that is rapidly customized for UPEC using E. coli UTI89 as a model strain. Through our rapid selection and combination of four phages against UPEC strain UTI89 (SR01, SR02, SR04, and Zappy) from our library, the combination of two lytic phages, SR02 and SR04, exhibits the strongest suppression of bacterial growth for at least 16 h, with no emergence of phage resistance observed in vitro. Phage SR02 undergoes subcellular activity for 25 min, producing approximately 106 progeny particles per cell, while SR04 completes its replication cycle in 20 min, generating around 564 progeny particles per cell. These two novel phages are genetically diverse, and their cocktail exhibited potent suppression of bacterial growth, independent of multiplicities of infection (MOIs), significantly reducing the viable bacterial counts after treatment in vitro. The phage cocktail has low immunogenicity and does not induce any proinflammatory gene responses in human bladder uroepithelial cells. Moreover, the cocktail effectively eradicates the invading UPEC strain UTI89 in the uroepithelial cells at a comparable level to that of phage SR04 alone, likely releasing some immunostimulatory agents that, in turn, trigger upregulation of MIP-3 and IL-8 genes. Altogether, this study offers an alternative pipeline for rapidly formulating genetically diverse phage-derived cocktails, which is specifically customized for targeted bacteria.

Chronic consumption of a high-fat diet (HFD) alters gut microbiota, induces endotoxemia, and impairs cognitive function (1, 2). Previous studies have shown that the administration of probiotics, such as Lactobacillus paracasei HII01, attenuated gut dysbiosis, alleviated metabolic disturbance, improved brain mitochondrial function, and improved cognitive function in obese insulin-resistant rats. However, the effects of probiotics lactobacillus reuteri KUB-AC5 on these parameters in obese insulin-resistant rats have not been investigated. Twenty-four male Wistar rats were fed either a normal diet (ND) or an HFD (59.28% energy from fat) for total 24 weeks. At week 13, each dietary group was then divided into subgroups receiving either vehicle or L. reuteri KUB-AC5 (1 ml daily oral feeding of 10 8 CFU) for an additional 12 weeks. At the end of the experimental protocol, the cognitive function was determined in all rats. Then animals were euthanized, and blood and brains were collected for further analysis. Chronic HFD consumption increased body weight, visceral fat weight, total cholesterol, insulin levels, increased HOMA-IR index, and increased serum malondialdehyde (MDA) level, when compared to ND-fed rats treated with vehicle. HFD-fed rats treated with probiotics significantly decreased body weight, insulin, serum MDA levels, and HOMA-IR index, when compared to HFD-fed rats treated with vehicle ( p  <0.05, Figure 1A-C). In addition, the impairment of brain mitochondrial function was observed in HFD-fed rats treated with vehicle by increasing percent change of brain ROS production when incubated with hydrogen peroxide, which was decreased in HFD-fed rats treated with probiotics ( p  <0.05, Figure 1D). Moreover, HFD-fed rats treated with vehicle decreased dendritic spine density and exhibited cognitive decline, which were attenuated in HFD-fed rats treated with probiotics ( p  <0.05, Figure 1E-F). We also observed that ND-fed rats treated with probiotics did not alter all parameters. Our findings suggest that the administration of L. reuteri KUB-AC5 mitigates cognitive decline in obese insulin-resistant rats by alleviating metabolic disturbances, reducing peripheral oxidative stress, improving brain mitochondrial function, and improving neuronal plasticity. These findings highlight its potential as a therapeutic intervention for metabolic and neurodegenerative disorders.

IntroductionBacterial urinary tract infections (UTI) are among the most common infectious diseases worldwide. The rise of multidrug-resistant (MDR) uropathogenic Escherichia coli (UPEC) UTI cases is a significant threat to healthcare systems. Several probiotic bacteria have been proposed as an alternative to combat MDR UTI. Lactic acid bacteria in the genus Limosilactobacillus are some of the most studied and used probiotics. However, strain-specific effects play a critical role in probiotic properties. L. reuteri KUB-AC5 (AC5), isolated from the chicken gut, confers antimicrobial and immunobiotic effects against some human pathogens. However, the antibacterial and immune modulatory effects of AC5 on UPEC have never been explored.MethodsHere, we investigated both the direct and indirect effects of AC5 against UPEC isolates (UTI89, CFT073, and clinical MDR UPEC AT31) in vitro. Using a spot-on lawn, agar-well diffusion, and competitive growth assays, we found that viable AC5 cells and cell-free components of this probiotic significantly reduced the UPEC growth of all strains tested. The human bladder epithelial cell line UM-UC-3 was used to assess the adhesion and pathogen-attachment inhibition properties of AC5 on UPEC.Results and discussionOur data showed that AC5 can attach to UM-UC-3 and decrease UPEC attachment in a dose-dependent manner. Pretreatment of UPEC-infected murine macrophage RAW264.7 cells with viable AC5 (multiplicity of infection, MOI = 1) for 24 hours enhanced macrophage-killing activity and increased proinflammatory (Nos2, Il6, and Tnfa) and anti-inflammatory (Il10) gene expression. These findings indicate the gut-derived AC5 probiotic could be a potential urogenital probiotic against MDR UTI.

We examined the activity of phages to control the growth of chicken and swine Salmonella strains in avian (CHIC-8E11), porcine (IPEC-1), and human (HT-29) cell cultures. We optimized a six-phage cocktail by selecting the five most effective myoviruses and a siphovirus that have optimal lysis on prevalent serovars. We observed ∼20% of 7 log10 PFU/well phage and 3-6 log10 CFU bacterial adhesions, and 3-5 log10 CFU bacterial invasion per 2 cm2 of the cultured cells at 2 h post-treatment. The invasive bacteria when plated had a variable reduced susceptibility to the phages. After phage application at an MOI of 10, the prophylaxis regimen had better efficacy at controlling bacterial growth with an up to 6 log10 CFU/well reduction as compared with the 1-2 log10 CFU/well bacterial reduction observed in the remedial and coinfection regimens. Our data support the development of these phages to control salmonellosis in chickens, pigs, and humans.

ABSTRACT Urinary tract infections are widespread bacterial infections affecting millions of people annually, with Escherichia coli being the most prevalent. Although phage therapy has recently gained interest as a promising alternative therapy for antibiotic-resistant bacteria, several studies have raised concerns regarding the evolution of phage resistance, making the therapy ineffective. In this study, we discover a novel coli myophage designated as Killian that targets E. coli strains, including the uropathogenic E. coli (UPEC) strain CFT073. It requires at least 20 minutes for 90% of its particles to adsorb to the host cells, undergoes subcellular activities for replication for 30 minutes, and eventually lyses the cells with a burst size of about 139 particles per cell. Additionally, Killian can withstand a wide variety of temperatures (4–50°C) and pHs (4 – 10). Genome analysis reveals that Killian’s genome consists of 169,905 base pairs with 35.5% GC content, encoding 276 open reading frames; of these, 209 are functionally annotated with no undesirable genes detected, highlighting its potential as an antibiotic alternative against UPEC. However, after an 8-hour phage treatment at high multiplicities of infection, bacterial density continuously increases, indicating an onset of bacterial growth revival. Thus, the combination study between the phage and three different antibiotics, including amikacin, ciprofloxacin, and piperacillin, was performed and showed that certain pairs of phage and antibiotics exhibited synergistic interactions in suppressing the bacterial growth revival. These findings suggest that Killian-antibiotic combinations are effective in inhibiting the growth of UPEC. IMPORTANCE Phage therapy has recently been in the spotlight as a viable alternative therapy for bacterial infections. However, several studies have raised concerns about the emergence of phage resistance that occurs during treatment, making the therapy not much effective. Here, we present the discovery of a novel E. coli myophage that, by itself, can effectively kill the uropathogenic E. coli, but the emergence of bacterial growth revival was detected during the treatment. Phage and antibiotics are then combined to improve the efficiency of the phage in suppressing the bacterial re-growth. This research would pave the way for the future development of phage-antibiotic cocktails for the sustainable use of phages for therapeutic purposes.

ABSTRACT In a looming post-antibiotic era, antibiotic alternatives have become key players in the combat against pathogens. Although recent advances in genomic research allow scientists to fully explore an organism’s genome in the search for novel antibacterial molecules, laborious work is still needed in order to dissect each individual gene product for its antibacterial activity. Here, we exploited phage-induced bacterial morphological changes as anchors to explore and discover a potential phage-derived antimicrobial embedded in the phage genome. We found that, upon vibriophage KVP40 infection, Vibrio parahaemolyticus exhibited morphological changes similar to those observed when treated with mecillinam, a cell wall synthesis inhibitor, suggesting the mechanism of pre-killing that KVP40 exerts inside the bacterial cell upon sieging the host. Genome analysis revealed that, of all the annotated gene products in the KVP40 genome that are involved in cell wall degradation, lytic transglycosylase (LT) is of particular interest for subsequent functional studies. A single-cell morphological analysis revealed that heterologous expression of wild-type KVP40-LT induced similar bacterial morphological changes to those treated with the whole phage or mecillinam, prior to cell burst. On the contrary, neither the morphology nor the viability of the bacteria expressing signal-peptide truncated- or catalytic mutant E80A- KVP40-LT was affected, suggesting the necessity of these domains for the antibacterial activities. Altogether, this research paves the way for the future development of the discovery of phage-derived antimicrobials that is guided through phage-induced morphological changes.

Perilla frutescens (PF) seed residue is a waste from perilla oil production that still contains nutrients and phytochemicals. This study aimed to investigate the chemoprotective action of PF seed residue crude ethanolic extract (PCE) on the inflammatory-induced promotion stage of rat colon carcinogenesis and cell culture models. PCE 0.1 and 1 g/kg body weight were administered by oral gavage to rats after receiving dimethylhydrazine (DMH) with one week of dextran sulfate sodium (DSS) supplementation. PCE at high dose exhibited a reduction in aberrant crypt foci (ACF) number (66.46%) and decreased pro-inflammatory cytokines compared to the DMH + DSS group (p < 0.01). Additionally, PCE could either modulate the inflammation induced in murine macrophage cells by bacterial toxins or suppress the proliferation of cancer cell lines, which was induced by the inflammatory process. These results demonstrate that the active components in PF seed residue showed a preventive effect on the aberrant colonic epithelial cell progression by modulating inflammatory microenvironments from the infiltrated macrophage or inflammatory response of aberrant cells. Moreover, consumption of PCE could alter rat microbiota, which might be related to health benefits. However, the mechanisms of PCE on the microbiota, which are related to inflammation and inflammatory-induced colon cancer progression, need to be further investigated.

Salmonella is a food-borne pathogen often linked to poultry sources, causing gastrointestinal infections in humans, with the numbers of multidrug resistant (MDR) isolates increasing globally. To gain insight into the genomic diversity of common serovars and their potential contribution to disease, we characterized antimicrobial resistance genes, and virulence factors encoded in 88 UK and 55 Thai isolates from poultry; the presence of virulence genes was detected through an extensive virulence determinants database compiled in this study. Long-read sequencing of three MDR isolates, each from a different serovar, was used to explore the links between virulence and resistance. To augment current control methods, we determined the sensitivity of isolates to 22 previously characterized Salmonella bacteriophages. Of the 17 serovars included, Salmonella Typhimurium and its monophasic variants were the most common, followed by S. Enteritidis, S. Mbandaka, and S. Virchow. Phylogenetic analysis of Typhumurium and monophasic variants showed poultry isolates were generally distinct from pigs. Resistance to sulfamethoxazole and ciprofloxacin was highest in isolates from the UK and Thailand, respectively, with 14-15% of all isolates being MDR. We noted that >90% of MDR isolates were likely to carry virulence genes as diverse as the srjF, lpfD, fhuA, and stc operons. Long-read sequencing revealed the presence of global epidemic MDR clones in our dataset, indicating they are possibly widespread in poultry. The clones included MDR ST198 S. Kentucky, harboring a Salmonella Genomic Island-1 (SGI)-K, European ST34 S. 1,4,[5],12:i:-, harboring SGI-4 and mercury-resistance genes, and a S. 1,4,12:i:- isolate from the Spanish clone harboring an MDR-plasmid. Testing of all isolates against a panel of bacteriophages showed variable sensitivity to phages, with STW-77 found to be the most effective. STW-77 lysed 37.76% of the isolates, including serovars important for human clinical infections: S. Enteritidis (80.95%), S. Typhimurium (66.67%), S. 1,4,[5],12:i:- (83.3%), and S. 1,4,12: i:- (71.43%). Therefore, our study revealed that combining genomics and phage sensitivity assays is promising for accurately identifying and providing biocontrols for Salmonella to prevent its dissemination in poultry flocks and through the food chain to cause infections in humans.