Yersinia Pseudotuberculosis Research Articles

Neutrophil inflammasomes sense the subcellular delivery route of translocated bacterial effectors and toxins.

In neutrophils, caspase-11 cleaves gasdermin D (GSDMD), causing pyroptosis to clear cytosol-invasive bacteria. In contrast, caspase-1 also cleaves GSDMD but seems to not cause pyroptosis. Here, we show that this pyroptosis-resistant caspase-1 activation is specifically programmed by the site of translocation of the detected microbial virulence factors. We find that pyrin and NLRC4 agonists do not trigger pyroptosis in neutrophils when they access the cytosol from endosomal compartment. In contrast, when the same ligands penetrate through the plasma membrane, they cause pyroptosis. Consistently, pyrin detects extracellular Yersinia pseudotuberculosis ΔyopM in neutrophils, driving caspase-1-GSDMD pyroptosis. This pyroptotic response drives PAD4-dependent H3 citrullination and results in extrusion of neutrophil extracellular traps (NETs). Our data indicate that caspase-1, GSDMD, or PAD4 deficiency renders mice more susceptible to Y.pseudotuberculosis ΔyopM infection. Therefore, neutrophils induce pyroptosis in response to caspase-1-activating inflammasomes triggered by extracellular bacterial pathogens, but after they phagocytose pathogens, they are programmed to forego pyroptosis.

Assessment of the Role of Free-Living and Farmed Fallow Deer (Dama dama) as A Potential Source of Human Infection with Multiple-Drug-Resistant Strains of Yersinia enterocolitica and Yersinia pseudotuberculosis.

Yersinia enterocolitica and Y. pseudotuberculosis are Gram-negative, facultative anaerobic bacteria that cause yersiniosis-one of the most important zoonotic diseases of the digestive tract. The aim of this study was to determine the prevalence of potentially human-pathogenic Y. enterocolitica and Y. pseudotuberculosis strains in free-living and farmed fallow deer, and to evaluate their sensitivity to chemotherapeutics. A total of 372 rectal swabs were analyzed, including 262 from free-living and 110 from farmed fallow deer. Due to the psychrophilic properties of Yersinia, two samples were collected from each animal. Seven Y. enterocolitica strains were isolated from free-living fallow deer, while two strains were isolated from farmed fallow deer. Yersinia pseudotuberculosis strains were not identified. All isolated Y. enterocolitica strains were ystB-positive, and phylogenetic analysis based on the nucleotide sequences of this gene revealed the presence of two phylogenetic groups. Yersinia enterocolitica strains isolated from fallow deer belonged to biotype 1A, and serotyping analysis demonstrated that the vast majority did not agglutinate with any diagnostic sera. All strains were multiple drug resistant and were not sensitive to at least four of the tested chemotherapeutics (amoxicillin with clavulanic acid, ampicillin, cefalexin, and streptomycin). One Y. enterocolitica strain isolated from a free-living animal was resistant to nine out of the 13 analyzed chemotherapeutics and was intermediately sensitive to the four remaining chemotherapeutics. The highest sensitivity was noted in case of ciprofloxacin (five strains) and trimethoprim-sulfamethoxazole (three strains). Only one strain isolated from a free-living animal was sensitive to three out of the 13 examined antibiotics, whereas the remaining strains were sensitive to only one drug or were not sensitive to any of the chemotherapeutics used. The results of this study indicate that multiple drug-resistant Y. enterocolitica strains can be carried by free-living and farmed fallow deer. This observation gives serious cause for concern because the meat of fallow deer and other ruminants is often consumed semi-raw (steak) or raw (tartar steak).

Injection-Site Sarcoma in Three Village Weaver Birds (Ploceus cucullatus) Associated with Autogenous Yersinia pseudotuberculosis Vaccination

Post-vaccinal sarcomas have been reported in cats and rarely in other domestic mammals, but not in birds. Three village weaver birds (Ploceus cucullatus) presented with poor flying ability and abnormal wing carriage attributable to large, unilateral pectoral masses. All had received at least one dose of autogenous Yersinia pseudotuberculosis vaccine into the affected pectoral muscle 74-408 days previously. Following euthanasia, gross post-mortem examination revealed locally invasive subcutaneous tumours extending through the sternum into the coelomic cavity. Cytology and histology revealed neoplasms of pleomorphic spindloid neoplastic cells with foci of coagulative necrosis and cavitation, sometimes containing faintly refractile non-polarizing granular material, both extracellularly and after phagocytosis by surrounding cells, including multinucleated giant cells. Immunohistochemistry in one bird supported a striated muscle cell origin. Findings of anaplastic sarcoma with intralesional foreign crystalline material resembled typical injection-site sarcomas in cats. This is the first report of presumptive vaccine-associated sarcoma in a non-mammalian species.

Gastroenterological masks of chronic yersiniosis in children: comprehension of experience

Chronic yersinioses are “therapeutic” infections with predominant symptoms of a somatic pathology while the typical manifestations of the infectious process are absent. Рurpose . Тo analyze the features of clinical manifestations of chronic yersinioses in children and to evaluate the eff ectiveness of diagnostic methods for detecting the disease. Characteristics of children and research methods. Laboratory confi rmation of the diagnosis of yersiniosеs was the criterion for including patients in the study group. As a result the group of patients with chronic yersiniosis infection included 131 children, 68 boys and 63 girls aged 2 years 8 months to 17 years 11 months (the average age of the group was 10 years) with typical gastroenterological pathology Laboratory confi rmation of the diagnosis of pseudotuberculosis and intestinal yersiniosis was carried out using the PCR to detect pathogenic yersinia in feces, specifi c antibodies were determined by a set of methods. Results . The article describes the clinical variants of chronic yersiniosis infections in children caused by Yersinia pseudotuberculosis and pathogenic Yersinia enterocolitica. Мost organs of the gastrointestinal tract (stomach, pancreas, liver, small and large intestines, mesenteric lymph nodes) can be aff ected in the course of chronic yersiniosis infection in children. A recurrent course of chronic yersiniosis infection is possible in children with hypogammaglobulinemia A. Conclusion . The chronic yersinioses can be suspected in the absence or insignifi cant eff ect of standard therapy of somatic pathology (gastritis, pancreatitis, irritable bowel syndrome, functional dyspepsia, etc.). The irritable bowel syndrome and functional dyspepsia are most common clinical variants of chronic yersiniosis infection in children. The immunoblotting is the most effective method to confi rm the diagnosis of chronic yersinioses.

The tempo and mode of gene regulatory programs during bacterial infection.

Innate immune recognition of bacterial pathogens is a key determinant of the ensuing systemic response, and host or pathogen heterogeneity in this early interaction can impact the course of infection. To gain insight into host response heterogeneity, we investigate macrophage inflammatory dynamics using primary human macrophages infected with Group B Streptococcus. Transcriptomic analysis reveals discrete cellular states within responding macrophages, one of which consists of four sub-states, reflecting inflammatory activation. Infection with six additional bacterial species-Staphylococcus aureus, Listeria monocytogenes, Enterococcus faecalis, Yersinia pseudotuberculosis, Shigella flexneri, and Salmonella enterica-recapitulates these states, though at different frequencies. We show that modulating the duration of infection and the presence of a toxin impacts inflammatory trajectory dynamics. We provide evidence for this trajectory in infected macrophages in an invivo model of Staphylococcus aureus infection. Our cell-state analysis defines a framework for understanding inflammatory activation dynamics in response to bacterial infection.

КВАНТОВЫЕ ТОЧКИ СУЛЬФИДА КАДМИЯ, ПОЛУЧЕННЫЕ С ИСПОЛЬЗОВАНИЕМ БЕЛКОВ-ПОРИНОВ, КАРРАГИНАНОВ, ХИТОЗАНОВ И ЛИПОПОЛИСАХАРИДОВ

Quantum dots (QDs) are a new generation of fluorochromes with significant advantages over traditional organic dyes. QDs based on CdS are promising materials for optics, optoelectronics, biology, and medicine. QDs in the form of colloidal solutions are of great scientific and practical interest. CdS quantum dots were synthesized by chemical condensation in an aqueous solution using Yersinia pseudotuberculosis porin proteins, positively (chitosan) and negatively (carrageenan, lipopolysaccharide) charged polysaccharides. The maxima of the emission spectra for all samples were 450 nm, which indicates the same QD size; is determined by the size of the "cells" of the grid matrix, which limit the size of the QD. It was shown that the fluorescence intensity of QDs synthesized in LPS solutions was two times higher than that of other samples. The fluorescence of the samples and the maxima of the emission spectra (450 nm) are preserved during intensive dialysis against buffers, which indicates the stability of QDs and the possibility of using the obtained labeled preparations. Keeping samples at pH 3 leads to a significant decrease in fluorescence, especially for acidic polysaccharides. Anionic oxygen of phosphate, hydroxyl groups of sugar, nitrogen atoms can interact with metal ions, which are precursors (precursors) for CdS nanocrystals. It was found that the interaction with porin-specific IgG leads to a significant change in the luminescence intensity of QD-porin samples. This is of interest from the point of view of chemical sensing and opens the prospect of using QD-labeled porin-based nanostructures as biosensors.

YERSINIA PSEUDOTUBERCULOSIS INFECTION IN LIONS (PANTHERA LEO) AT A ZOOLOGICAL PARK.

Two co-housed 17-yr-old male lions (Panthera leo) and one 15-yr-old female lion in an adjacent enclosure developed acute lethargy, depression, anorexia, and ataxia at a zoological park in central North Carolina, United States. One of the male lions and the female lion were found dead 5 and 4 d after the onset of clinical signs, respectively. The other male lion recovered without any clinical treatment. A third male lion housed with the female lion never developed clinical signs. Postmortem examination, microscopic evaluation, and bacterial culture of the liver from both deceased lions confirmed systemic Yersinia pseudotuberculosis infection. Susceptibility testing revealed resistance to amoxicillin-clavulanic acid and cefazolin. To investigate the extent of the outbreak, qualitative real-time polymerase chain reaction (PCR) for Y. pseudotuberculosis was performed on feces and substrate from 15 enclosures housing nondomestic felids and canids, resulting in a positive sample from one enclosure housing four asymptomatic lions. This enclosure was adjacent to housing of the deceased female lion. Enrofloxacin was administered to all animals in the vicinity at doses of 5 mg/kg in nondomestic felids and 10 mg/kg in nondomestic canids, orally q24h for 14 d. Repeated fecal PCR performed 1 wk after completing the antibiotic treatment protocol found no positive samples. The source of the infection was not identified despite PCR testing of environmental samples from all enclosures in the vicinity, the remains of a prey item fed out prior to the outbreak, and a single dead rodent found on grounds. No further clinical cases have occurred within the following year.

Chronic Leptin Deficiency Improves Tolerance of Physiological Damage and Host-Pathogen Cooperation during Yersinia pseudotuberculosis Infection.

To combat infections, hosts employ a combination of antagonistic and cooperative defense strategies. The former refers to pathogen killing mediated by resistance mechanisms, while the latter refers to physiological defense mechanisms that promote host health during infection independent of pathogen killing, leading to an apparent cooperation between the host and the pathogen. Previous work has shown that Leptin, a pleiotropic hormone that plays a central role in regulating appetite and energy metabolism, is indispensable for resistance mechanisms, while a role for Leptin signaling in cooperative host-pathogen interactions remains unknown. Using a mouse model of Yersinia pseudotuberculosis (Yptb) infection, an emerging pathogen that causes fever, diarrhea, and mesenteric lymphadenitis in humans, we found that the physiological effects of chronic Leptin-signaling deficiency conferred protection from Yptb infection due to increased host-pathogen cooperation rather than greater resistance defenses. The protection against Yptb infection was independent of differences in food consumption, lipolysis, or fat mass. Instead, we found that the chronic absence of Leptin signaling protects from a shift to lipid utilization during infection that contributes to Yptb lethality. Furthermore, we found that the survival advantage conferred by Leptin deficiency was associated with increased liver and kidney damage. Our work reveals an additional level of complexity for the role of Leptin in infection defense and demonstrates that in some contexts, in addition to tolerating the pathogen, tolerating organ damage is more beneficial for survival than preventing the damage.

Climate Changes in Central Asia as a Prerequisite and Trigger of Plague Microbe (Yersinia pestis) Speciation.

Two physical and climatic factors that were distant and recent prerequisites and a transformation trigger for a clone of the ancestral pseudotuberculous microbe Yersinia pseudotuberculosis O:1b (the causative agent of the Far East scarlet-like fever (FESLF)) into a population of the plague microbe derivative Y. pestis are considered. One remote prerequisite was the aridification of the Central Asian landscapes in the second half of the Cenozoic period and the formation of the Gobi Arid Zone. The arid conditions of Central Asia determined the formation of adaptive species-specific protective behavior in the Tarbagan marmot (Marmota sibirica) when installing the plug of a wintering hole, which later contributed to the massive infection of the animals with FESLF by the aberrant (traumatic, not alimentary) method during hibernation. A recent prerequisite and a real trigger of Y. pestis speciation was the onset of the last maximum (Sartan) ice age in Central Asia at the turn of the Pleistocene and Holocene, 22 000–15 000 years ago. Freezing of the cooling burrows of the Tarbagan marmot caused a behavioral shift in the larval population of the marmot flea Oropsylla silantiewi and the transition to the cold winter–spring months of the year from saprophagy in the nesting litter to hematophagy on the bodies of sleeping animals. Larval scarifications in the oral cavity of sleeping marmots have become the entrance gate for a unique traumatic FESLF infection. The constellation of climatic changes, the heterothermal (and, accordingly, heteroimmune) condition of the family groups of sleeping marmots, and the year-round propagation of marmot fleas in wintering burrows, combined with behavioral shifts in marmots and fleas caused by climatic changes, led to the formation of conditions in the parasitic system M. sibirica–O. silantiewi in which the transformation of the FESLF microbe into the causative agent of the plague occurred according to peripatric speciation. Thus, the climatic changes that happened at different times in the Cenozoic initially led to a shift in behavior of the Tarbagan marmot and, subsequently, to a shift in the behavior of the fleas parasitizing it. Ultimately, the change in the behavior of marmots and fleas caused the transition of the clone(s) of the FESLF causative agent into a new ecological niche and adaptive zone, as well as the transformation into a population(s) of the plague microbe.

Protection Induced by Oral Vaccination with a Recombinant Yersinia pseudotuberculosis Delivering Yersinia pestis LcrV and F1 Antigens in Mice and Rats against Pneumonic Plague.

A newly attenuated Yersinia pseudotuberculosis strain (designated Yptb1) with triple mutation Δasd ΔyopK ΔyopJ and chromosomal insertion of the Y. pestis caf1R-caf1M-caf1A-caf1 operon was constructed as a live vaccine platform. Yptb1 tailored with an Asd+ plasmid (pYA5199) (designated Yptb1[pYA5199]) simultaneously delivers Y. pestis LcrV and F1. The attenuated Yptb1(pYA5199) localized in the Peyer's patches, lung, spleen, and liver for a few weeks after oral immunization without causing any disease symptoms in immunized rodents. An oral prime-boost Yptb1(pYA5199) immunization stimulated potent antibody responses to LcrV, F1, and Y. pestis whole-cell lysate (YPL) in Swiss Webster mice and Brown Norway rats. The prime-boost Yptb1(pYA5199) immunization induced higher antigen-specific humoral and cellular immune responses in mice than a single immunization did, and it provided complete short-term and long-term protection against a high dose of intranasal Y. pestis challenge in mice. Moreover, the prime-boost immunization afforded substantial protection for Brown Norway rats against an aerosolized Y. pestis challenge. Our study highlights that Yptb1(pYA5199) has high potential as an oral vaccine candidate against pneumonic plague.

The Assessment on Synergistic Activity of Ebselen and Silver Ion Against Yersinia pseudotuberculosis.

Yersinia pseudotuberculosis is a foodborne zoonotic bacterium that is pathogenic to guinea pigs, rabbits, and mice. It also causes pseudotuberculosis in humans. However, it still lacked the scientific basis for control. Here, we found out that Ebselen (EbSe) exhibited synergistic antibacterial activity with silver nitrate (Ag+) against Y. pseudotuberculosis YpIII strain with high efficacy in vitro using UV-visible light absorption spectrum, 5,5’-dithiobis-(2-nitrobenzoic acid), laser scanning confocal microscope, flow cytometry, transmission electron microscopy and Western blotting assays. The depletion of total glutathione (GSH) amount and inhibition of thioredoxin reductase (TrxR) activity in thiol-dependent redox system revealed the destructiveness of EbSe-Ag+-caused intracellular oxidative stress. Furthermore, a YpIII-caused mice gastroenteritis model was constructed. EbSe-Ag+ significantly reduced bacterial loads with low toxicity. It also down-regulated the expression levels of interferon (IL)-1β and tumor necrosis factor-α, up-regulated the expression level of IL-10 on-site. All the in vivo results demonstrated the antibacterial activity and immune-modulatory property of EbSe-Ag+. Collectively, these results provided academic fundament for further analysis and development of EbSe-Ag+ as the antibacterial agents for pseudotuberculosis control.

Investigation of salicylidene acylhydrazides derivatives: Molecular Docking, ADMET, and Molecular Dynamic Simulations were used in conjunction towards the design of new Yersinia pseudotuberculosis inhibitors

LysR-type transcription factor RovM is an important target of Yersinia pseudotuberculosis drug discovery and the discovery of antibacterial is considered one of the greatest medical achievements of all time. In this research work, a combination of three docking tools with different algorithms was applied in Salicylidene acylhydrazides derivatives intended toward gram-negative bacterium Yersinia pseudotuberculosis to evaluate their binding interactions.
 The analysis of the molecular docking results obtained from the 3-docking software system succeeded in screening twelve fascinating compounds with higher restrictive concentrations having a decent affinity to LysR-type transcription factor RovM macromolecule. Then the Lipinski’s and Veber’s rule properties were calculated to spot the drug-likeness properties of the investigated candidate compounds. To anticipate the toxicity of the predicted candidate chemicals, in-silico toxicity tests were conducted. Furthermore, golden triangle and drug scores were performed, the investigated compounds which fall within the golden triangle indicate that these compounds would not have clearance problems. 5 of the 12 hits drugs pass the golden triangle screening step. These selected drugs undergo a drug score test which only compound 17 passed. To validate the stability, 1 ns molecular dynamic simulations were done on the highest-ranking drug score compound 17 / 3onm complexes. These findings point to interesting avenues for the development of new compounds that are more effective against Yersinia pseudotuberculosis.

Quinolone‐Rhodanine Hybrid Compounds: Synthesis and Biological Evaluation as Anti‐Bacterial Agents

AbstractAs potential antimicrobial agents, 4‐fluoroaniline and diethyl (ethoxymethylene)malonate were used to efficiently synthesize a new quinolone (3‐9) series. The 1HNMR and 13C NMR, IR, mass spectrometry, and elemental analysis of all compounds were used to identify them. The final compounds 3–9 were in vitro screened for antimicrobial activity against gram negative‐gram positive Escherichia coli, Yersinia pseudotuberculosis, Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus faecalis, Bacillus cereus, Mycobacterium smegmatis and fungal strains Candida albicans, Saccharomyces cerevisiae. The best antimicrobial activity was demonstrated by compounds 9a and 9b. Antimicrobial resistance and microbial infection could benefit from quinolone derivatives′ ability to inhibit the growth of microorganisms, according to this study.

RNA Thermometer-coordinated Assembly of the Yersinia Injectisome

The type III secretion system (T3SS) is indispensable for successful host cell infection by many Gram-negative pathogens. The molecular syringe delivers effector proteins that suppress the host immune response. Synthesis of T3SS components in Yersinia pseudotuberculosis relies on host body temperature, which induces the RNA thermometer (RNAT)-controlled translation of lcrF coding for a virulence master regulator that activates transcription of the T3SS regulon. The assembly of the secretion machinery follows a strict coordinated succession referred to as outside-in assembly, in which the membrane ring complex and the export apparatus represent the nucleation points. Two components essential for the initial assembly are YscJ and YscT. While YscJ connects the membrane ring complex with the export apparatus in the inner membrane, YscT is required for a functional export apparatus. Previous transcriptome-wide RNA structuromics data suggested the presence of unique intercistronic RNATs upstream of yscJ and yscT. Here, we show by reporter gene fusions that both upstream regions confer translational control. Moreover, we demonstrate the temperature-induced opening of the Shine-Dalgarno region, which facilitates ribosome binding, by in vitro structure probing and toeprinting methods. Rationally designed thermostable RNAT variants of the yscJ and yscT thermometers confirmed their physiological relevance with respect to T3SS assembly and host infection. Since we have shown in a recent study that YopN, the gatekeeper of type III secretion, also is under RNAT control, it appears that the synthesis, assembly and functionality of the Yersinia T3S machinery is coordinated by RNA-based temperature sensors at multiple levels.

Horizontal Transfer of Virulence Factors by Pathogenic Enterobacteria to Marine Saprotrophic Bacteria during Co-Cultivation in Biofilm.

Environmental problems associated with marine pollution and climate warming create favorable conditions for the penetration and survival of pathogenic bacteria in marine ecosystems. These microorganisms have interspecific competitive interactions with marine bacteria. Co-culture, as an important research strategy that mimics the natural environment of bacteria, can activate silent genes or clusters through interspecies interactions. The authors used modern biotechnology of co-cultivation to dynamically study intercellular interactions between different taxa of bacteria—pathogenic enterobacteria Yersinia pseudotuberculosis and Listeria monocytogenes and saprotrophic marine bacteria Bacillus sp. and Pseudomonas japonica isolated in summer from the coastal waters of the recreational areas of the Sea of Japan. The results of the experiments showed that during the formation of polycultural biofilms, horizontal transfer of genes encoding some pathogenicity factors from Y. pseudotuberculosis and L. monocytogenes to marine saprotrophic bacteria with different secretion systems is possible. It was previously thought that this was largely prevented by the type VI secretion system (T6SS) found in marine saprotrophic bacteria. The authors showed for the first time the ability of marine bacteria Bacillus sp. and P. japonica to biofilm formation with pathogenic enterobacteria Y. pseudotuberculosis and L. monocytogenes, saprophytic bacteria with type III secretion system (T3SS). For the first time, a marine saprotrophic strain of Bacillus sp. Revealed manifestations of hyaluronidase, proteolytic and hemolytic activity after cultivation in a polycultural biofilm with listeria. Saprotrophic marine bacteria that have acquired virulence factors from pathogenic enterobacteria, including antibiotic resistance genes, could potentially play a role in altering the biological properties of other members of the marine microbial community. In addition, given the possible interdomain nature of intercellular gene translocation, acquired virulence factors can be transferred to marine unicellular and multicellular eukaryotes. The results obtained contribute to the paradigm of the epidemiological significance and potential danger of anthropogenic pollution of marine ecosystems, which creates serious problems for public health and the development of marine culture as an important area of economic activity in coastal regions.

Yersinia pseudotuberculosis doxycycline tolerance strategies include modulating expression of genes involved in cell permeability and tRNA modifications.

Antibiotic tolerance is typically associated with a phenotypic change within a bacterial population, resulting in a transient decrease in antibiotic susceptibility that can contribute to treatment failure and recurrent infections. Although tolerant cells may emerge prior to treatment, the stress of prolonged antibiotic exposure can also promote tolerance. Here, we sought to determine how Yersinia pseudotuberculosis responds to doxycycline exposure, to then verify if these gene expression changes could promote doxycycline tolerance in culture and in our mouse model of infection. Only four genes were differentially regulated in response to a physiologically-relevant dose of doxycycline: osmB and ompF were upregulated, tusB and cnfy were downregulated; differential expression also occurred during doxycycline treatment in the mouse. ompF, tusB and cnfy were also differentially regulated in response to chloramphenicol, indicating these could be general responses to ribosomal inhibition. cnfy has previously been associated with persistence and was not a major focus here. We found deletion of the OmpF porin resulted in increased antibiotic accumulation, suggesting expression may promote diffusion of doxycycline out of the cell, while OsmB lipoprotein had a minor impact on antibiotic permeability. Overexpression of tusB significantly impaired bacterial survival in culture and in the mouse, suggesting that tRNA modification by tusB, and the resulting impacts on translational machinery, promotes survival during treatment with an antibiotic classically viewed as bacteriostatic. We believe this may be the first observation of bactericidal activity of doxycycline under physiological conditions, which was revealed by reversing tusB downregulation.

Outbreak of Yersinia pseudotuberculosis in capybaras (Hydrochoerus hydrochaeris) kept in captivity

On January 2020, an outbreak of acute mortality was detected in the capybara (Hydrochoerus hydrochaeris) population kept in the Cordoba Zoo Conservation Center (CZCC) in Spain. Morbidity and mortality rates were 71%, and the fatality rate was 100%. Macroscopic examination revealed multifocal nodular lesions with fibrinonecrotic exudate in lungs and mesenteric lymph nodes. Microscopic lesions were characterized as severe acute fibrinopurulent bronchopneumonia and generalized vascular changes. Cultures and biochemical analyses yielded Yersinia pseudotuberculosis (YPT). This species was confirmed by both PCR and MALDI-TOF. This report describes the first outbreak of fatal YPT infection in zoo animals in Spain and confirms that capybaras are highly susceptible to this pathogen, which could be of animal health, conservation, and public health concern. Future studies are warranted to determine potential sources of YTP infection in zoos to prevent future cases in susceptible species, including humans.

Osteomyelitis in a slaughter turkey flock caused by Yersinia pseudotuberculosis sequence type ST42

A Yersinia pseudotuberculosis outbreak was diagnosed in a male turkey flock in Finland. Y. pseudotuberculosis is a quite rare zoonotic bacterium, which typically causes enteritis in humans and sudden death in animals. In this study, osteomyelitis was diagnosed in small, lame, 11- to 12-wk-old male turkeys. Lameness and slower growth among the turkeys was observed on the farm. During pathological examination, multiple lesions were found in the metaphyseal and physeal areas of the femurs, tibiotarsi, and tarsometatarsi, with multifocal to coalescing mixed heterophilic/granulomatous necrotizing osteomyelitis. Y. pseudotuberculosis was isolated from the femoral and tibiotarsal bones or from the joints of six lame turkeys sent for necropsy. The isolation required homogenizing of lesion tissue in phosphate-mannitol-peptone broth, which was cultured directly - and, if needed, after cold enrichment – on selective cefsulodin-irgasan-novobiocin agar. Whole-genome sequencing was used for identification and typing. All isolates belonged to bio/serotype 1/O:1a and sequence type ST42 (Achtman scheme), which is commonly reported in both human and animal Y. pseudotuberculosis infections in Europe. The isolates from all six turkeys showed only one to two allele differences in the core genome comparison, indicating a common source of infection. All asymptomatic turkeys were slaughtered at the age of 17 weeks. Whole and partial carcass condemnation rates at the slaughterhouse were high, but no macroscopic changes in the skeletal system were found, showing that food chain information is essential. This study confirms earlier findings that Y. pseudotuberculosis can cause osteomyelitis in fattening turkeys, leading to lameness. Food chain information is essential for slaughterhouse operations, to protect the workers and emphasize good working hygiene during slaughter.

Adaptation Potential of Three Psychrotolerant Aquatic Bacteria in the Pan-Okhotsk Region

The Pan-Okhotsk region, which is part of the western North Pacific Ocean, is famous for its active volcanoes, which are part of the Pacific Ring of Fire and that enrich the surrounding waters with essential chemicals. Therefore, this region, including the Sea of Okhotsk and the Sea of Japan, is characterized by rich biota. Bacterioplankton plays a significant part in biological communities and is an indicator of ecosystem function. Analyzing the adaptability of three representatives of the microbiota of the Pan-Okhotsk region was the goal of our investigation. Marinomonas primoryensis KMM3633T (MP), Yersinia ruckeri KMM821 (YR), and Yersinia pseudotuberculosis 598 (YP) from the G.B. Elyakov Pacific Institute of Bioorganic Chemistry were studied by means of genomic and bioinformatic methods. The list of membrane translocator proteins, metabolism pathways, and cold shock and antifreeze proteins that were revealed in the genome of MP characterized this bacterium as being adaptable to free living in marine conditions, even at winter temperatures. The genomic potential of YR and YP makes not only survival in the environment of the Pan-Okhotsk region but also pathogenesis in eukaryotic organisms possible. The data obtained will serve as a basis for further ecosystem monitoring with the help of microbiota research.

Cpx-signalling facilitates Hms-dependent biofilm formation by Yersinia pseudotuberculosis

Bacteria often reside in sessile communities called biofilms, where they adhere to a variety of surfaces and exist as aggregates in a viscous polymeric matrix. Biofilms are resistant to antimicrobial treatments, and are a major contributor to the persistence and chronicity of many bacterial infections. Herein, we determined that the CpxA-CpxR two-component system influenced the ability of enteropathogenic Yersinia pseudotuberculosis to develop biofilms. Mutant bacteria that accumulated the active CpxR~P isoform failed to form biofilms on plastic or on the surface of the Caenorhabditis elegans nematode. A failure to form biofilms on the worm surface prompted their survival when grown on the lawns of Y. pseudotuberculosis. Exopolysaccharide production by the hms loci is the major driver of biofilms formed by Yersinia. We used a number of molecular genetic approaches to demonstrate that active CpxR~P binds directly to the promoter regulatory elements of the hms loci to activate the repressors of hms expression and to repress the activators of hms expression. Consequently, active Cpx-signalling culminated in a loss of exopolysaccharide production. Hence, the development of Y. pseudotuberculosis biofilms on multiple surfaces is controlled by the Cpx-signalling, and at least in part this occurs through repressive effects on the Hms-dependent exopolysaccharide production.