Different characteristics of the soil in marmot habitats might be one of the factors that influcting Yersinia pestis prevalent in which than pikas.

Introduction: Marmots are recognized as host animals for plague caused by Yersinia pestis infection. It is unclear that why plague prevalent in marmot rather than other rodents like pikas in the same habitats. This study aims to analyze the differences of the soil characteristics around marmots and pikas burrows to explore the soils factors impacting on different epidemic intensities of Yersinia pestis in these two rodents. Methods: Soil samples were collected from within and around marmot and pika burrows, as well as from the nearby areas not inhabited by them and Chinese baseline soil properties as control groups, in the Qilian Mountains of Gansu Province, China. The physicochemical properties and the bacterial 16S rRNA were measured to analyze the characteristics of soils from different groups. Subsequently, the data were analyzed using R studio. Results: The analysis revealed that marmot habitats exhibited distinct soil characteristics, including lower organic matter and alkaline hydrolyzed nitrogen, but higher electrical conductivity and total soluble salts. And soil in marmot areas tended to have higher concentrations of nickel, chromium, and iron, also lower levels of zinc and selenium. Additionally, the alpha diversity of soil microorganisms in marmot habitats was significantly low. Simultaneously, redundancy analysis was conducted, which showed that the low alpha diversity of marmot-soil was influenced by its physicochemical properties. The alpha diversity of the soil was positively correlated with EC, TSS, Na, and Cr, etc., while it was negatively correlated with AHN, OM, Se, Zn, and Fe, etc. Conclusion: These characteristics in marmot habitats, including low levels of organic matter, alkaline hydrolyzed nitrogen, zinc, selenium, and bacterial alpha diversity, as well as high levels of electrical conductivity, total soluble salts, iron, and nickel, played a crucial role in the spread of plague. It was discovered that the unique characteristics of marmot-soils provided essential elements necessary for the survival of Yersinia pestis, including high levels of Fe and Ca, or facilitated the spread of plague. Thus, the transmission of the plague was facilitated.

Neutralizing Chimeric Anti-F1 Monoclonal Antibody against Yersinia pestis Infection.

Drug-resistant Yersinia pestis (Y. pestis) poses a threat to the use of antibiotics to treat Y. pestis infections. Passive immunization with neutralizing monoclonal antibodies (mAbs) is one approach to the treatment of infectious diseases. In this study, a murine single-chain fragment variable (scFv) phage antibody library targeting the F1 antigen was constructed and screened. Therapeutic intravenous injection of 400 µg chimeric mAb S1 through tail veins provided complete protection against Y. pestis (strain: 91001) challenge in a pneumonic plague mouse model. Timely antibody treatment eliminated the bacteria and reduced lung inflammation. Our data suggest that chimeric mAb S1 is a candidate treatment for Y. pestis infection that warrants further study.

An investigative study on Yersinia enterocolitica in animals, humans and dried milk in New Valley Governorate, Egypt.

BACKGROUND: Yersiniosis is one of the most significant intestinal disorders caused by Yersinia enterocolitica and affects both humans and animals. This study aimed to investigate the prevalence of Y. enterocolitica in New Valley Governorate, Egypt in animals, humans, fresh milk and dried milk. Additionally, this study analyzed the presence of virulence genes, including ail and Yst in tested isolates and conducted a phylogenetic analysis to determine the genetic similarity between human, and animal Y. enterocolitica isolates. Finally, the antimicrobial resistance patterns of the isolates were examined. RESULTS: Among the 982 samples examined, the prevalence of Y. enterocolitica based on ISO10273-2017 was 11.7% in animal samples including 12.8% of animal faeces, and 10.4% in milk samples. Moreover, the prevalence of Y. enterocolitica was 13.2% in human stool, and 9.5% in dried milk samples. The molecular characterization of the six randomly selected isolates showed that the 16S rRNA, ail and Yst genes were found in 50, 33.3 and 100% of the examined Y. enterocolitica isolates, respectively. Phylogenetic analysis of animal and human isolates based on the 16S rRNA gene revealed a high degree of similarity between the isolates. All the tested animal and human Y. enterocolitica isolates (100%) were resistant to ampicillin and cefotaxime, but highly sensitive to norfloxacin. CONCLUSIONS: The high prevalence of Y. enterocolitica in animal and human samples with high degrees of genetic similarity poses a threat to public and animal health. Animal faeces, milk and milk powder represent the main sources of Y. enterocolitica infection in humans. Additionally, high levels of antibiotic resistance of Y. enterocolitica can cause public health hazards by leading to the failure of disease prevention and treatment programs in humans and animals.

In silico vaccine design for Yersinia enterocolitica: A comprehensive approach to enhanced immunogenicity, efficacy and protection.

Yersinia enterocolitica, a foodborne pathogen, has emerged as a significant public health concern due to its increased prevalence and multidrug resistance. This study employed reverse vaccinology to identify novel vaccine candidates against Y. enterocolitica through comprehensive in silico analyses. The core genome's conserved protein translocase subunit SecY was selected as the target, and potential B-cell, MHC class I, and MHC class II epitopes were mapped. 3B-cell epitopes, 3 MHCI and 11 MHCII epitopes were acquired. A multi-epitope vaccine construct was designed by incorporating the identified epitopes, TLR4 Agonist was used as adjuvants to enhance the immunogenic response. EAAAK, CPGPG and AYY linkers were used to form a vaccine construct, followed by extensive computational evaluations. The vaccine exhibited desirable physicochemical properties, stable secondary and tertiary structures as evaluated by PDBSum and trRosetta. Moreover, favorable interactions with the human Toll-like receptor 4 (TLR4) was observed by ClusPro. Population coverage analysis estimated the vaccine's applicability across 99.74 % in diverse populations. In addition, molecular dynamics simulations and normal mode analysis confirmed the vaccine's structural stability and dynamics in a simulated biological environment. Furthermore, codon optimization and in silico cloning facilitated the evaluation of the vaccine's expression potential in E. coli and pET-28a was used a recombinant plasmid. This study provides a promising foundation for the development of an efficacious vaccine against Y. enterocolitica infections.

Stability assessment of housekeeping genes for qRT-PCR in Yersinia enterocolitica cultured at 22°C and 37°C.

Yersinia enterocolitica, a species within the genus Yersinia, thrives optimally at 22-25°C but can also grow at the mammalian core body temperature of 37°C. This dual temperature adaptability necessitates establishing both temperature conditions in research to examine the effects on various biological processes. In quantitative real-time PCR (qRT-PCR) assays, the selection of appropriate housekeeping genes is vital for data accuracy. Nevertheless, the lack of alternatives and information often leads to the default use of the 16S rRNA gene despite potential limitations. This investigation sourced 16 potential reference genes through a comprehensive review of the literature and transcriptome sequencing data analysis. We validated the expression stability of these genes via qRT-PCR across 12 Y. enterocolitica strains, representing the four prevalent serotypes O:3, O:5,27, O:8, and O:9, isolated from diarrheal patient stool samples. This approach aimed to minimize the impact of serotype heterogeneity. After acquiring Cq values, gene stability was evaluated using four established algorithms-ΔCq, geNorm, NormFinder, and BestKeeper-and subsequently synthesized into a consolidated ranking through the Robust Rank Aggregation (RRA) method. Our study suggests that the genes glnS, nuoB, glmS, gyrB, dnaK, and thrS maintain consistent expression across varying culture temperatures, supporting their candidacy as robust housekeeping genes. We advise against the exclusive use of 16S rRNA for this purpose. Should tradition prevail in its utilization, it must be employed with discernment, preferably alongside one or two of the housekeeping genes identified in this study as internal controls.IMPORTANCEIn our study, we focused on identifying stable reference genes for quantitative real-time PCR (qRT-PCR) experiments on Y. enterocolitica cultured at different temperatures (22°C and 37°C). After thoroughly evaluating 16 candidate genes, we identified six genes-glnS, nuoB, glmS, gyrB, dnaK, and thrS-as exhibiting stable expression across these temperature conditions, making them ideal reference genes for Y. enterocolitica studies. This discovery is crucial for ensuring the accuracy and reliability of qRT-PCR data, as the choice of appropriate reference genes is key to normalizing expression data and minimizing experimental variability. Importantly, our research extended beyond bioinformatics analysis by incorporating validation with clinical strains, bridging the gap between theoretical predictions and practical application. This approach not only underscores the robustness and reliability of our findings but also directly addresses the critical need for experimental validation in the field. By providing a set of validated, stably expressed reference genes, our work offers valuable guidance for designing experiments involving Y. enterocolitica, enhancing the reliability of research outcomes, and advancing our understanding of this significant pathogen.

Status and analysis of undetected plague cases in Yunnan Province, China.

Background: The virulence of Yersinia pestis strains in the Rattus flavipectus plague focus is relatively low. The purpose of this study was to investigate the undetected, sporadic plague cases in plague foci and provide the basis for plague prevention and control. Methods: A 3-year-old plague-confirmed case was investigated in the R. flavipectus plague focus of Yunnan Province in 2020 due to the intensive screening for fever symptoms during the coronavirus disease 2019 (COVID-19) pandemic. Epidemiological investigation, laboratory testing, and clinical treatment were conducted for the case. The expanded survey was carried out around the case within a 7-km radius, including the resident population, domesticated dogs, and rats. PCR and indirect hemagglutination tests were performed on the collected samples. Results: The isolation rates of Y. pestis were 100.0% (7 out of 7) in dead rats and 4.00% (3 out of 75) in live rats in the survey area of the foci. A total of 5.00% (6 out of 120) of the domesticated dogs were F1 antibody positive. Nine local people were determined for plague infection recently (0.92%, 9 out of 978). The locations of human cases coincided with the Y. pestis epidemic area among the animals. Conclusion: This study discovered the existence of plague cases that had not been detected by routine surveillance in the R. flavipectus plague focus, and the actual epidemic of human infection may be underestimated.

The Putative Virulence Plasmid pYR4 of the Fish Pathogen Yersinia ruckeri Is Conjugative and Stabilized by a HigBA Toxin-Antitoxin System.

The bacterium Yersinia ruckeri causes enteric redmouth disease in salmonids and hence has substantial economic implications for the farmed fish industry. The Norwegian Y. ruckeri outbreak isolate NVH_3758 carries a relatively uncharacterized plasmid, pYR4, which encodes both type 4 pili and a type 4 secretion system. In this study, we demonstrate that pYR4 does not impose a growth burden on the Y. ruckeri host bacterium, nor does the plasmid contribute to twitching motility (an indicator of type 4 pilus function) or virulence in a Galleria mellonella larval model of infection. However, we show that pYR4 is conjugative. We also reveal, through mutagenesis, that pYR4 encodes a functional post-segregational killing system, HigBA, that is responsible for plasmid maintenance within Y. ruckeri. This is the first toxin-antitoxin system to be characterized for this organism. Whilst further work is needed to elucidate the virulence role of pYR4 and whether it contributes to bacterial disease under non-laboratory conditions, our results suggest that the plasmid possesses substantial stability and transfer mechanisms that imply importance within the organism. These results add to our understanding of the mobile genetic elements and evolutionary trajectory of Y. ruckeri as an important commercial pathogen, with consequences for human food production.

Microbiological Quality and Antibiotic Resistance of Relevant Bacteria from Horsemeat.

The aim of this work was to assess the microbiological safety and quality of horsemeat. A total of 19 fresh horsemeat samples were analysed. Mesophile counts were 4.89 ± 1.08 log CFU/g, and Enterobacteriaceae, Staphylococcus spp., and enterococci were only isolated from 36.84%, 21.05%, and 15.79% of the samples, respectively. Neither Staphylococcus aureus nor Escherichia coli were found in any sample. Listeria spp. and Listeria monocytogenes were detected in 31.58% and 21.05% of the samples, respectively. Campylobacter jejuni was not detected in any sample. The dominant bacteria were lactic acid bacteria. Seven different Staphylococcus spp. were identified, the most common being S. delphini, S. saprophyticus, and S. warneri. S. delphini showed resistance against mupirocin and cefoxitin. All the L. monocytogenes strains showed resistance against ampicillin, cefotaxime, and oxacillin. Multi-resistant Yersinia enterocolitica, Stenotrophomonas maltophilia, and Vagococcus. fluvialis strains were found, with resistance to 11, 7, and 8 antibiotics, respectively, causing significant concern. Therefore, specific actions should be taken to decrease the contamination of horsemeat.

Preliminary Study of the Characterization of the Viable but Noncultivable State of Yersinia enterocolitica Induced by Chloride and UV Irradiation.

The viable but non-culturable (VBNC) state is a survival strategy for many foodborne pathogens under adverse conditions. Yersinia enterocolitica (Y. enterocolitica) as a kind of primary foodborne pathogen, and it is crucial to investigate its survival strategies and potential risks in the food chain. In this study, the effectiveness of ultraviolet (UV) irradiation and chlorine treatment in disinfecting the foodborne pathogen Y. enterocolitica was investigated. The results indicated that both UV irradiation and chlorine treatment can induce the VBNC state in Y. enterocolitica. The bacteria completely lost culturability after being treated with 25 mg/L of NaClO for 30 min and a UV dose of 100 mJ/cm². The number of culturable and viable cells were detected using plate counting and a combination of fluorescein and propidium iodide (live/dead cells). Further research found that these VBNC cells exhibited reduced intracellular Adenosine Triphosphate (ATP) levels, and increased levels of reactive oxygen species (ROS) compared to non-induced cells. Morphologically, the cells changed from a rod shape to a shorter, coccobacillary shape with small vacuoles forming at the edges, indicating structural changes. Both condition-induced VBNC-state cells were able to resuscitate in tryptic soy broth (TSB) medium supplemented with Tween 80, sodium pyruvate, and glucose. These findings contribute to a better understanding of the survival mechanisms of Y. enterocolitica in the environment and are of significant importance for the development of effective disinfection strategies.

Disseminated Yersinia enterocolitica infection associated to Sweet's syndrome.

We report the case of a young woman sough care for disseminated form of Yersinia enterocolitica infection (pseudoappendicitis with mesenteric lymph node, arthralgia, glomerulonephritis and hepatitis) diagnosed on Western Blot method for the detection of Yersinia antibodies. The patient also presented a rare concomitant cutaneous manifestation, as Sweet's syndrome, confirmed histologically. Neutrophilic dermatosis is an exceptional skin features among post-infectious autoimmune disorders when encountering Yersinia enterocolitica infection in clinical practice.

Screening of promising molecules against potential drug targets in Yersinia pestis by integrative pan and subtractive genomics, docking and simulation approach.

This study focuses on Yersinia pestis, the bacterium responsible for plague, which posed a severe threat to public health in history. Despite the availability of antibiotics treatment, the emergence of antibiotic resistance in this pathogen has increased challenges of controlling the infections and plague outbreaks. The development of new drug targets and therapies is urgently needed. This research aims to identify novel protein targets from 28 Y. pestis strains by the integrative pan-genomic and subtractive genomics approach. Additionally, it seeks to screen out potential safe and effective alternative therapies against these targets via high-throughput virtual screening. Targets should lack homology to human, gut microbiota, and known human 'anti-targets', while should exhibit essentiality for pathogen's survival and virulence, druggability, antibiotic resistance, and broad spectrum across multiple pathogenic bacteria. We identified two promising targets: the aminotransferase class I/class II domain-containing protein and 3-oxoacyl-[acyl-carrier-protein] synthase 2. These proteins were modeled using AlphaFold2, validated through several structural analyses, and were subjected to molecular docking and ADMET analysis. Molecular dynamics simulations determined the stability of the ligand-target complexes, providing potential therapeutic options against Y. pestis.

Plasmonic-Enhanced Colorimetric Lateral Flow Immunoassays Using Bimetallic Silver-Coated Gold Nanostars.

The colorimetric lateral flow immunoassay (cLFIA) has gained widespread attention as a point-of-care testing (POCT) technique due to its low cost, short analysis time, portability, and capability of being performed by unskilled operators with minimal requirement of reagents. However, the low analytical sensitivity of conventional LFIA based on colloidal gold nanospheres limits their applications for sensitive detection of trace amounts of target analytes. In this study, we introduced a novel plasmonic-enhanced colorimetric LFIA (PE-cLFIA) platform featuring bimetallic silver-coated gold nanostars (BGNS) with exceptional optical properties, leading to ultrahigh visual color brightness. The BGNS-based PE-cLFIA was successfully applied to detect a model analyte, low-calcium response V (LcrV), a virulence protein factor found in Yersinia pestis, the causative agent of bubonic plague. The PE-cLFIA sensing using BGNS-3 composed of 45 nm silver thickness showed a high visual colorimetric sensitivity with a detection limit as low as 13.7 pg/mL, which was around 50 times more sensitive than that of a traditional gold nanoparticle-based LFIA. In addition, the antibody-conjugated BGNS-3 showed excellent stability over 6 months. To illustrate the potential for clinical applications, we demonstrated that our LFIA platform for detecting LcrV spiked in human serum without any sample preprocessing exhibited a detection limit of 22.8 pg/mL. These results open up new opportunities for developing hybrid nanoparticle systems for sensitive POCT PE-cLFIA screening for infectious disease detection.

A case-control study and molecular epidemiology of yersiniosis in Aotearoa New Zealand.

The objective of this study was to determine risk factors and sources attributed to yersiniosis in Aotearoa New Zealand (NZ). A risk factor questionnaire was administered to 247 notified yersiniosis cases and 258 control participants from the Canterbury and/or Wellington regions of NZ. Yersinia sp. isolates from clinical cases and a range of food sources were whole-genome sequenced and genetically compared. Yersinia enterocolitica (YE) bioserotype 2/3, O:9 [McNally multi-locus sequence type (ST) 12] and YE Biotype (BT) 1A (46 different STs) predominated within the consented cases (45 and 27%, respectively). Exposure to pork was identified as a significant risk factor for cases associated with YE ST12. The presence of YE ST12 was confirmed in retail raw meat, primarily raw pork. Single-nucleotide polymorphism (SNP) analysis identified multiple genomically very closely related clusters (0-5 SNPs) of YE ST12, predominately from raw pork with clinical cases from one or both regions. Risk factors associated with YE BT 1A included the consumption of cooked seafood, sushi, tofu, and some vegetable types. Analysis of specific risk factors and SNP analysis, combined, indicate that raw pork is a significant risk factor for exposure and infection to pathogenic YE cases, but not BT 1A cases.

Differential expression of the type III secretion system genes in Yersinia ruckeri: Preliminary investigations in different environmental conditions.

Type III secretion system (T3SS) is an important virulence system in Gram-negative bacteria. In this investigation, different environmental conditions that regulate the expression of T3SS genes in Yersinia ruckeri were investigated aimed at obtaining a better understanding about its modulation after various environmental challenges. Four isolates of Y. ruckeri CSF007-82, ATCC29473, A7959-11 and YRNC10 were cultivated under the diverse in vitro challenges iron depletion, high salt, low pH and in the presence of fish serum or in the fish cell culture (Chinook Salmon Embryo - CHSE). The transcriptional modulation of the chromosomal genes ysaV, ysaC, ysaJ and prgH of ysa were investigated using quantitative real-time PCR. The expression of prgH, ysaV, ysaC and ysaJ was differentially expressed in all four strains under evaluation. The highest gene expression levels were observed for Y. ruckeri YRNC10 AN after addition of 0.3 M NaCl in Luria Bertani broth. The results obtained from this study provide initial insights into T3SS responses in Y. ruckeri, which pave the way for further studies aimed at expanding our knowledge on the functional roles of the T3SS genes in Y. ruckeri.

Multiplex PCR System for the Diagnosis of Plague.

The plague caused by Yersinia pestis has a high case fatality rate. It is often transmitted from person to person through mosquito bites, causing serious disease transmission. Due to its clinical symptoms being very similar to influenza, it is difficult to detect by people. Traditional detection methods for Y. pestis mainly include bacterial culture and serological identification, which are cumbersome and require high experimental conditions. Therefore, a fast and effective detection method is very important. At present, polymerase chain reaction (PCR) is one of the methods for rapid detection of Y. pestis. In this review, we focus on the application, advantages, and disadvantages of multiplex PCR technology in clinical detection.

Biomarkers of oxidative stress, biochemical changes, and the activity of lysosomal enzymes in the livers of rainbow trout (Oncorhynchus mykiss Walbaum) vaccinated against yersiniosis before a Yersinia ruckeri challenge.

Introduction: This study aimed to evaluate biomarkers of oxidative stress (2-thiobarbituric acid reactive substances, aldehyde and ketone derivatives of oxidatively modified proteins and total antioxidant capacity), the activity of antioxidant enzymes (superoxide dismutase, catalase, glutathione reductase and glutathione peroxidase), that of lysosomal enzymes (alanyl aminopeptidase, leucyl aminopeptidase, ß-N-acetylglucosaminidase and acid phosphatase) and changes in biochemical parameters (alanine aminotransferase, aspartate aminotransferase, de Ritis ratio, lactate dehydrogenase activity, lactate and pyruvate levels and their ratio) in the liver tissue of fish that were vaccinated against enteric redmouth disease and challenged with its causative agent, the bacterium Yersinia ruckeri. Material and Methods: The vaccine was administered orally to trout, some of which were challenged with Y. ruckeri 61 days later. For comparison, unvaccinated and unchallenged trout and unvaccinated and challenged trout were also evaluated. Results: In the unvaccinated fish, infection with Y. ruckeri disrupted the pro-oxidant/antioxidant balance, led to a significant increase in lipid peroxidation and oxidative modification of proteins, decreased total antioxidant capacity and significantly increased the activity of lysosomal enzymes. In vaccinated fish, the Y. ruckeri challenge increased the activity of glutathione-related enzymes and decreased lipid peroxidation, anaerobic metabolism and the activity of lysosomal enzymes in fish livers relative to the unvaccinated and challenged group. In contrast, these parameters increased after the Y. ruckeri challenge in unvaccinated trout relative to those in the untreated group. Conclusion: Vaccination exerted a protective effect during the Y. ruckeri challenge and had no adverse effect on fish livers.

Detection of Gastrointestinal Pathogens with Zoonotic Potential in Horses Used in Free-Riding Activities during a Countrywide Study in Greece.

The objectives of this study were (a) to detect zoonotic gastrointestinal pathogens in faecal samples of horses using the FilmArray® GI Panel and (b) to identify variables potentially associated with their presence. Faecal samples collected from 224 horses obtained during a countrywide study in Greece were tested by means of the BioFire® FilmArray® Gastrointestinal (GI) Panel, which uses multiplex-PCR technology for the detection of 22 pathogens. Gastrointestinal pathogens were detected in the faecal samples obtained from 97 horses (43.3%). Zoonotic pathogens were detected more frequently in samples from horses in courtyard housing (56.0%) than in samples from horses in other housing types (39.7%) (p = 0.040). The most frequently detected zoonotic pathogens were enteropathogenic Escherichia coli (19.2% of horses) and Shiga-like toxin-producing E. coli stx1/stx2 (13.8%). During multivariable analysis, two variables emerged as significant predictors for the outcome 'detection of at least one zoonotic pathogen in the faecal sample from an animal': (a) the decreasing age of horses (p = 0.0001) and (b) the presence of livestock at the same premises as the horses (p = 0.013). As a significant predictor for the outcome 'detection of two zoonotic pathogens concurrently in the faecal sample from an animal', only the season of sampling of animals (autumn) emerged as significant in the multivariable analysis (p = 0.049). The results indicated a diversity of gastrointestinal pathogens with zoonotic potential in horses and provided evidence for predictors for the infections; also, they can serve to inform horse owners and handlers regarding the possible risk of transmission of pathogens with zoonotic potential. In addition, our findings highlight the importance of continuous surveillance for zoonotic pathogens in domestic animals.

Unveiling Milan's hidden cases of plague occurred in autumn 1629, before the great 1630 epidemic.

In the summer of 1630, Milan experienced the most devastating plague epidemic in its history. In this study, addressed to investigate the earliest phases of the epidemic in the autumn of 1629, a set of unpublished and only partially known primary sources produced by the city's Officium Sanitatis was consulted and compared for the first time. Including those of two foreigners who died in the Lazzaretto, it was possible to ascertain a total of 39 cases of plague occurred in Milan between 9 October 1629 and the first weeks of 1630, of which 29 (74.4%) ended in death. Seven deaths presumably occurred at home were not recorded in the Liber Mortuorum, in which at least three other deaths caused by plague were deliberately attributed to a different cause. In particular, the case of the Vicario di Provisione in charge, Alfonso Visconti, probably the first death from plague occurred in Milan that year, was deliberately concealed for political reasons. Nevertheless, the spread of the disease remained limited in autumn 1629 and it was probably stopped until the following spring more by climatic factors than by the interventions of public health officials.

Identification of the organic peroxide scavenging system of Yersinia pseudotuberculosis and its regulation by OxyR.

Oxidative stress caused by reactive oxygen species (ROS) is inevitable for all aerobic microorganisms as ROS are the byproducts of aerobic respiration. For gut pathogens, ROS are an integrated part of colonization resistance which protects the host against bacteria invasion. Alkyl hydroperoxide reductase (AhpR) and organic hydroperoxide resistance (Ohr) proteins are considered as the main enzymes responsible for the degradation of organic peroxides (OPs) in most bacteria. To elucidate how enteric pathogen Yersinia pseudotuberculosis YPIII deals with oxidative stress induced by OPs, we performed transcriptomic analysis and identified the OP scavenging system, which is composed of glutathione peroxidase (Gpx), thiol peroxidase (Tpx), and AhpR. Gpx serves as the main scavenger of OPs, and Tpx assists in the degradation of OPs. Transcriptional factor OxyR regulates Gpx expression, suggesting that OxyR is the regulator mediating the cellular response to OPs. Although AhpR has little influence on OP degradation, its deletion would greatly impair the scavenging ability of OPs in the absence of gpx or tpx. In addition, we found that catalase KatG and KatE are responsive to OPs but do not participate in the removal of OPs.IMPORTANCEIn bacteria, oxidative stress caused by ROS is a continuously occurring cellular response and requires multiple genes to participate in this process. The elimination of OPs is mainly dependent on AhpR and Ohr protein. Here, we carried out transcriptomic analysis to search for enzymes responsible for the removal of organic peroxides in Yersinia pseudotuberculosis. We found that Gpx was the primary OP scavenger in bacteria, which was positively regulated by the oxidative stress regulator OxyR. The OP scavenging system in Y. pseudotuberculosis was composedof Gpx, Tpx, and AhpR. OxyR is the critical global regulator mediating gene expression involved in OPs and H2O2 stress. These findings suggest that Y. pseudotuberculosis has a unique defense system in response to oxidative stress.

A bacteriophage cocktail targeting Yersinia pestis provides strong post-exposure protection in a rat pneumonic plague model.

Yersinia pestis, one of the deadliest bacterial pathogens ever known, is responsible for three plague pandemics and several epidemics, with over 200 million deaths during recorded history. Due to high genomic plasticity, Y. pestis is amenable to genetic mutations as well as genetic engineering that can lead to the emergence or intentional development of pan-drug-resistant strains. Indeed, antibiotic-resistant strains (e.g., strains carrying multidrug-resistant or MDR plasmids) have been isolated in various countries and endemic areas. Thus, there is an urgent need to develop novel, safe, and effective treatment approaches for managing Y. pestis infections. This includes infections by antigenically distinct strains for which vaccines (none FDA approved yet) may not be effective and those that cannot be managed by currently available antibiotics. Lytic bacteriophages provide one such alternative approach. In this study, we examined post-exposure efficacy of a bacteriophage cocktail, YPP-401, to combat pneumonic plague caused by Y. pestis CO92. YPP-401 is a four-phage preparation effective against a panel of at least 68 genetically diverse Y. pestis strains. Using a pneumonic plague aerosol challenge model in gender-balanced Brown Norway rats, YPP-401 demonstrated ~88% protection when delivered 18 h post-exposure for each of two administration routes (i.e., intraperitoneal and intranasal) in a dose-dependent manner. Our studies provide proof-of-concept that YPP-401 could be an innovative, safe, and effective approach for managing Y. pestis infections, including those caused by naturally occurring or intentionally developed multidrug-resistant strains.IMPORTANCECurrently, there are no FDA-approved plague vaccines. Since antibiotic-resistant strains of Y. pestis have emerged or are being intentionally developed to be used as a biothreat agent, new treatment modalities are direly needed. Phage therapy provides a viable option against potentially antibiotic-resistant strains. Additionally, phages are nontoxic and have been approved by the FDA for use in the food industry. Our study provides the first evidence of the protective effect of a cocktail of four phages against pneumonic plague, the most severe form of disease. When treatment was initiated 18 h post infection by either the intranasal or intraperitoneal route in Brown Norway rats, up to 87.5% protection was observed. The phage cocktail had a minimal impact on a representative human microbiome panel, unlike antibiotics. This study provides strong proof-of-concept data for the further development of phage-based therapy to treat plague.