Yersinia Enterocolitica Research Articles

Biological and genomic characterization of three psychrophilic Y. enterocolitica phages.

Yersinia (Y.) enterocolitica is an important foodborne pathogenic species that is mainly transmitted by the consumption of contaminated meat, particularly pork. To combat the bacteria along the food chain, the application of strictly lytic phages may be a promising tool. As the temperatures in the gut of animals and during food processing can differ significantly, a phage cocktail intended to be used for applications should comprise phages that are active at various temperatures. In this study, we isolated and characterized three phages with a myoviridal morphology (vB_YenM_P8, vB_YenM_P744 and vB_YenM_P778), which lysed the most important Y. enterocolitica serotypes O:3, O:9 and O:5,27 at a low multiplicity of infection (MOI) and at low temperatures down to 6°C. While vB_YenM_P8 is a member of the T4 family, vB_YenM_P744 and vB_YenM_P778 are novel phages that do not show relationship to known phages. The three phages were mixed in a cocktail with the already described phages vB_YenM_P281 and vB_YenP_Rambo. The cocktail revealed a strong lytic activity and lysed a mixture of Y. enterocolitica serotypes at room temperature (RT) within few hours with a reduction of up to 4.8 log10 units. Moreover, at even lower temperatures the mixture was significantly reduced after incubation overnight. The strongest reductions were determined at 6°C (4.0 log10 units) suggesting that the cocktail can lyse the psychrophilic Y. enterocolitica also during food processing. To determine possible phage resistance, 100 colonies that survived the infection by the phages were isolated and analysed regarding their serotype and phage susceptibility. Most isolates belonged to serotype O:9, but all of them were still sensitive to at least one phage of the cocktail.

Yersinia enterocolitica biovar 1A: An underappreciated potential pathogen in the food chain

Yersinia enterocolitica is an underreported cause of foodborne gastroenteritis. Little is known of the diversity of Y. enterocolitica isolated from food and which food commodities contribute to human disease. In this study, Y. enterocolitica was isolated from 37/50 raw chicken, 8/10 pork, 8/10 salmon and 1/10 leafy green samples collected at retail in the UK. Up to 10 presumptive Y. enterocolitica isolates per positive sample underwent whole genome sequencing (WGS) and were compared with publicly available genomes. In total, 207 Y. enterocolitica isolates were analyzed and belonged to 38 sequence types (STs). Up to five STs of Y. enterocolitica were isolated from individual food samples and isolates belonging to the same sample and ST differed by 0–74 single nucleotide polymorphisms (SNPs). Biotype was predicted for 205 (99 %) genomes that all belonged to biotype 1A, previously described as non-pathogenic. However, around half (51 %) of food samples contained isolates belonging to the same ST as previously isolated from UK human cases. The closest human-derived isolates shared between 17 and 7978 single nucleotide polymorphisms (SNPs) with the food isolates. Extensive food surveillance is required to determine what food sources are responsible for Y. enterocolitica infections and to re-examine the role of biotype 1A as a human pathogen.

FARKLI KURUTMA TEKNİKLERİNİN HURMA ERİĞİ MEYVELERİNİN (DIOSPYROS LOTUS L.) BİYOAKTİF, RENK, ANTİBAKTERİYEL VE DUYUSAL ÖZELLİKLERİ ÜZERİNE ETKİSİ

In this study, the effects of hot air drying (HD), vacuum drying (VD) and freeze drying (FD) processes on total phenolic (TPC) and flavonoid contents (TFC), antioxidant capacity (AC), color and antibacterial properties and sensory evaluation of date plum (DP) fruit were determined. Total phenolic, total flavonoid and antioxidant capacity values of dried and fresh samples were determined as 13.86-7.45 mg GAE/g, 7.17-4.09 mg CE/g and 86.60-54.98%, respectively. The highest inhibition levels against Bacillus cereus (24.50 mm) and Salmonella Typhimurium (24.67 mm), Yersinia enterocolitica (25.75 mm) and Staphylococcus aureus (25.17 mm) were determined for HD and VD, respectively. In terms of sensory evaluation, FD drying was scored similarly to fresh samples (P>0.05). Overall, FD drying could be applied for longer preservation of DP fruits in terms of bioactive properties, color and sensory evaluation.

Use of recombinant malate dehydrogenase (MDH) and superoxide dismutase (SOD) [Cu[sbnd]Zn] as antigens in indirect ELISA for diagnosis of bovine brucellosis

The objective of this study was to validate an indirect enzyme-linked immunoassay (iELISA) using the recombinant proteins, malate dehydrogenase (MDH) and superoxide dismutase (SOD) [CuZn], as antigens and to evaluate its ability to discriminate antibodies produced by vaccination from those induced by infection in the diagnosis of bovine brucellosis. Sera from six groups were evaluated: G1 – culture-positive animals (52 serum samples) (naturally infected); G2 - non-vaccinated animals (28 serum samples) positive in RBT (Rose Bengal test) and 2ME (2-mercaptoethanol test) selected from brucellosis-positive herds; G3 - animals from a brucellosis-free area (32 serum samples); G4 - S19 vaccinated heifers (114 serum samples); G5 – RB51 vaccinated heifers (60 serum samples); G6 - animals inoculated with inactivated Yersinia enterocolitica O:9 (42 serum samples). Diagnostic sensitivity (DSe) and diagnostic specificity (DSp) were estimated using the frequentist approach and the confidence interval (CI) (95%) calculated by the Clopper-Pearson (exact) method. The DSe for iELISA_MDH in the G1 group was 71.7% (CI 95%: 57.6–83.2%) and for the G2 100.0% (CI 95%: 87.7–100.0%), whereas the DSp was 84.4% in the G3 (CI 95%: 67.2–94.7%). For the iELISA_SOD the DSe was estimated 67.3% for the G1 (CI 95%: 52.9–79.7%) and 71.4% for G2 (CI 95%: 51.3–86.8%), while the DSp for G3 was 87.5% (CI 95%: 71.0–96.5%). iELISA_MDH and iELISA_SOD showed potential to be used in the diagnosis of infected animals, increasing the range of serological tests available for the diagnosis of bovine brucellosis, with the advantage of being S-LPS-free. However, none of the tests could differentiate between infection and vaccination.

Characterization of microbial contamination in agricultural soil: A public health perspective

Soil is widely recognized as a reservoir of microbial contaminants including antibiotic resistance genes (ARGs) and human bacterial pathogens (HBPs), which are major public health concerns. Although the risks associated with soil safety in different soil habitats have been studied, the results are not comprehensive. In this study, dryland soils used for vegetable, corn, and soybean planting, and submerged soils used for rice planting and crab farming were collected and subjected to metagenomic sequencing to characterize HBPs, ARGs, and virulence factor genes (VFGs). The results showed that submerged soils had a higher abundance of HBP than dryland soils. In addition, the submerged soil microbiome acquired significantly higher levels of high-risk ARGs than the dryland soil microbiome and these ARGs were mainly assigned to bacA, sul1, and aadA genes submerged. Network analysis revealed that 11 HBPs, including Yersinia enterocolitica, Vibrio cholerae, Escherichia coli, and Leptospira interrogans, were high-risk because of their close association with ARGs, VFGs, and mobile genetic elements (MGEs). Procrustes and network analyses showed that HBPs and ARGs were more closely linked in submerged soil. This study confirms that submerged field has higher ecological environment risk and human health risk than dryland soil.

Comparison of Yersinia enterocolitica DNA Methylation at Ambient and Host Temperatures.

Pathogenic bacteria recognize environmental cues to vary gene expression for host adaptation. Moving from ambient to host temperature, Yersinia enterocolitica responds by immediately repressing flagella synthesis and inducing the virulence plasmid (pYV)-encoded type III secretion system. In contrast, shifting from host to ambient temperature requires 2.5 generations to restore motility, suggesting a link to the cell cycle. We hypothesized that differential DNA methylation contributes to temperature-regulated gene expression. We tested this hypothesis by comparing single-molecule real-time (SMRT) sequencing of Y. enterocolitica DNA from cells growing exponentially at 22 °C and 37 °C. The inter-pulse duration ratio rather than the traditional QV scoring was the kinetic metric to compare DNA from cells grown at each temperature. All 565 YenI restriction sites were fully methylated at both temperatures. Among the 27,118 DNA adenine methylase (Dam) sites, 42 had differential methylation patterns, while 17 remained unmethylated regardless of the temperature. A subset of the differentially methylated Dam sites localized to promoter regions of predicted regulatory genes including LysR-type and PadR-like transcriptional regulators and a cyclic-di-GMP phosphodiesterase. The unmethylated Dam sites localized with a bias to the replication terminus, suggesting they were protected from Dam methylase. No cytosine methylation was detected at Dcm sites.

One-step preparation of a self-assembled bioconjugate nanovaccine against Brucella

ABSTRACT Brucellosis, caused by Brucella, is a severe zoonosis, and the current Brucella live attenuated vaccine cannot be used in humans due to major safety risks. Although polysaccharide antigens can be used to prepare the Brucella vaccine, their lower immunogenicity limits them from producing efficient and broad protection. In this study, we produced a high-performance bioconjugate nanovaccine against different species of Brucella by introducing a self-assembly nanoparticle platform and an O-linked glycosylation system into Yersinia enterocolitica serotype O:9, which has an O-polysaccharide composed of the same unit as Brucella. After successfully preparing the vaccine and confirming its stability, we subsequently demonstrated the safety of the vaccine in mice by high-dose immunization. Then, by a series of mouse experiments, we found that the nanovaccine greatly promoted antibody responses. In particular, the increase of IgG2a was more obvious than that of IgG1. Most importantly, this nanovaccine could provide cross-protection against B. abortus, B. melitensis, and B. suis strains by lethal dose challenged models, and could improve the clearance of B. melitensis, the most common pathogenic species in human brucellosis, by non-lethal dose infection. Overall, for the first time, we biocoupled polysaccharide antigens with nano carriers to prepare a Brucella vaccine, which showed pronounced and extensive protective effects in mice. Thus, we provided a potential candidate vaccine and a new direction for Brucella vaccine design.

Preliminary Data on Escherichia coli, Yersinia enterocolitica, and Other Bacteria, as Well as Absent African Swine Fever Virus in the Gut Microbiota of Wild Mice and Voles from Bulgaria

Small mammals are bioindicator organisms, and, through their gut microbiota (GM), could be carriers of pathogens and resistant bacteria. Also, wild GM composition has been suggested to have large implications for conservation efforts. Seventeen bacterial species were obtained from intestinal samples of Bulgarian yellow-necked mice (Apodemus flavicollis) and bank voles (Myodes glareolus) via classic microbiological cultivation and biochemical identification. Twelve Gram-negative—Escherichia coli, Yersinia enterocolitica, Yersinia kristensenii, Hafnia alvei, Serratia liquefaciens, Serratia marcescens, Serratia proteamaculans, Pseudescherichia vulneris, Klebsiella pneumoniae ssp. ozaenae, Enterobacter cloacea, Pantoea agglomerans, Pseudomonas fluorescens group—and five Gram-positive bacteria, Enterococcus faecium, Enterococcus faecalis, Enterococcus hirae, Bacillus thuringiensis, and Lysinibacillus sphaericus, were discovered. Enterobacteriaceae was the most abundant family. The isolates belonged to one of the major reported taxa in rodents, Firmicutes (the Gram-positive species) and to the less abundant, but still among the first, phyla, Proteobacteria (the Gram-negative strains). We did not find any members of the other major phylum, Bacteroidetes, likely due to lack of metagenomic techniques. E. coli and Y. enterocolitica were confirmed with polymerase chain reaction. Almost all strains had pathogenic potential, but the good condition of the test animals suggests their commensal role. The Y. enterocolitica strains did not have the ail pathogenicity gene. There was high prevalence of multi-drug resistance (MDR), but for the expected species with high level of intrinsic resistance, such as the enterococci and S. marcescens. E. coli and some other species had very low antimicrobial resistance (AMR), in line with other studies of wild rodents. Many of the strains had biotechnological potential; e.g., B. thuringiensis is the most used biological insecticide, with its proteins incorporated into the Bt genetically modified maize. The GM of the tested wild mice and voles from Bulgaria proved to be a source of bacterial diversity; many of the strains were promising in terms of biotechnology, and, in addition, the samples did not contain the African swine fever virus.

Profiling of Microbial Landscape in Lung of Chronic Obstructive Pulmonary Disease Patients Using RNA Sequencing.

The aim of the study was to use RNA sequencing (RNA-seq) data of lung from chronic obstructive pulmonary disease (COPD) patients to identify the bacteria that are most commonly detected. Additionally, the study sought to investigate the differences in these infections between normal lung tissues and those affected by COPD. We re-analyzed RNA-seq data of lung from 99 COPD patients and 93 non-COPD smokers to determine the extent to which the metagenomes differed between the two groups and to assess the reliability of the metagenomes. We used unmapped reads in the RNA-seq data that were not aligned to the human reference genome to identify more common infections in COPD patients. We identified 18 bacteria that exhibited significant differences between the COPD and non-COPD smoker groups. Among these, Yersinia enterocolitica was found to be more than 30% more abundant in COPD. Additionally, we observed difference in detection rate based on smoking history. To ensure the accuracy of our findings and distinguish them from false positives, we double-check the metagenomic profile using Basic Local Alignment Search Tool (BLAST). We were able to identify and remove specific species that might have been misclassified as other species in Kraken2 but were actually Staphylococcus aureus, as identified by BLAST analysis. This study highlighted the method of using unmapped reads, which were not typically used in sequencing data, to identify microorganisms present in patients with lung diseases such as COPD. This method expanded our understanding of the microbial landscape in COPD and provided insights into the potential role of microorganisms in disease development and progression.

Comparative Genomics, Phylogenetic and Functional Analysis of Yersinia enterocolitica, a Gastrointestinal Pathogen, with Other Soil-Borne Bacteria Causing Diseases

Yersinia enterocolitica is a harmful bacterium transmitted through contaminated food, causing gastrointestinal illness and lymph node inflammation. The rise of drug-resistant strains of Y. enterocolitica poses a serious public health threat, necessitating research on its ecology, related species, and unique genes linked to virulence and antibiotic resistance. This study identified eight microorganisms similar to Y. enterocolitica and conducted a pan-genomic analysis, revealing specific genes exclusive to Y. enterocolitica. Enrichment analysis of these genes unveiled their involvement in antibiotic synthesis pathways, such as siderophore production, osmoregulated periplasmic glucan activation, and antibiotic resistance. These pathways, including biofilm formation and increased antibiotic tolerance, are vital for Yersinia’s virulence. Furthermore, specific genes related to glutamate metabolism, nitrogen regulation, motility, purine, and pyrimidine synthesis may contribute to Y. enterocolitica’s pathogenicity, growth, and virulence factor production. Phylogenetic analysis demonstrated the evolutionary relationship between Y. enterocolitica and similar species like Escherichia coli, Campylobacter jejuni, and Salmonella enterica, stressing the need to monitor Y. enterocolitica in slaughterhouses due to animal carriers. The study’s findings shed light on the ecological factors and genetic mechanisms driving Y. enterocolitica’s pathogenicity and antibiotic resistance. Targeting genes involved in purine and pyrimidine synthesis, such as ushA, cpdB, and deoB, could be potential strategies for controlling pathogenicity and antimicrobial resistance. Understanding the relationships and genetic interactions between Y. enterocolitica and related microorganisms is crucial for developing effective surveillance and management approaches in the future.

Detection of multiple human enteropathogens in Norway rats (Rattus norvegicus) from an under-resourced neighborhood of Vancouver, British Columbia.

Urban Norway rats (Rattus norvegicus) can carry various human pathogens, and may be involved in pathogen propagation and transmission to humans. From January 31-August 14, 2021, a community outbreak of Shigella flexneri serotype 2a occurred among unhoused or poorly housed people in the Downtown Eastside neighborhood of Vancouver, British Columbia, Canada. The source could not be identified; however, patients reported contact with rats, and previous studies indicated transmission of rat-associated zoonotic pathogens among the unhoused or poorly housed residents of this neighborhood. The study objective was to determine if rats trapped in the outbreak area were carriers of Shigella spp. and other zoonotic enteric pathogens. From March 23-April 9, 2021, 22 rats were lethally trapped within the outbreak area. Colonic content was analyzed using the BioFire FilmArray Gastrointestinal (multiplex PCR) panel for human enteropathogens, which detected: Campylobacter spp. (9/22), Clostridioides difficile (3/22), Yersinia enterocolitica (5/22), Cryptosporidium spp. (8/22), Giardia duodenalis (5/22), Rotavirus A (1/22), enteroaggressive Escherichia coli (2/22), enteropathogenic E. coli (10/22), and Shigella spp. or enteroinvasive E. coli (EIEC) (3/22). An ipaH PCR assay was used for targeted detection of Shigella spp./EIEC, with five rats positive. Two samples contained insertion sites unique to S. flexneri isolated from the human outbreak. This study highlights the potential for rats to carry a broad range of human pathogens, and their possible role in pathogen maintenance and/or transmission.

CRISPR-Cas12a test strip (CRISPR/CAST) package: In-situ detection of Brucella from infected livestock

BackgroundBrucellosis is a common zoonotic disease caused by Brucella, which causes enormous economic losses and public burden to epidemic areas. Early and precise diagnosis and timely culling of infected animals are crucial to prevent the infection and spread of Brucella. In recent years, RNA-guided CRISPR/Cas12a(Clustered Regularly Interspaced Short Palindromic Repeats and its associated protein 12a) nucleases have shown great promise in nucleic acid detection. This research aims to develop a CRISPR/CAST (CRISPR/Cas12a Test strip) package that can rapidly detect Brucella nucleic acid during on-site screening, especially on remote family pastures. The CRISPR/Cas12a system combined with recombinase polymerase amplification (RPA), and lateral flow read-out.ResultsWe selected the conserved gene bp26, which commonly used in Brucella infection detection and compared on Genbank with other Brucella species. The genomes of Brucella abortus 2308, Brucella suis S2, Brucella melitansis 16 M, and Brucella suis 1330, et al. were aligned, and the sequences were found to be consistent. Therefore, the experiments were only performed on B. melitensis. With the CRISPR/CAST package, the assay of Brucella nucleic acid can be completed within 30 min under isothermal temperature conditions, with a sensitivity of 10 copies/μl. Additionally, no antigen cross-reaction was observed against Yersinia enterocolitica O:9, Escherichia coli O157, Salmonella enterica serovar Urbana O:30, and Francisella tularensis. The serum samples of 398 sheep and 100 cattle were tested by the CRISPR/CAST package, of which 31 sheep and 8 cattle were Brucella DNA positive. The detection rate was consistent with the qPCR results and higher than that of the Rose Bengal Test (RBT, 19 sheep and 5 cattle were serum positive).ConclusionsThe CRISPR/CAST package can accurately detect Brucella DNA in infected livestock within 30 min and exhibits several advantages, including simplicity, speed, high sensitivity, and strong specificity with no window period. In addition, no expensive equipment, standard laboratory, or professional operators are needed for the package. It is an effective tool for screening in the field and obtaining early, rapid diagnoses of Brucella infection. The package is an efficient tool for preventing and controlling epidemics.

Drinking Pipes and Nipple Drinkers in Pig Abattoir Lairage Pens-A Source of Zoonotic Pathogens as a Hazard to Meat Safety.

The water distribution system in the lairage pens of abattoirs could act as a route of contamination for produced meat. In this study, biofilm formation and the occurrence of specific pathogens in drinking equipment was investigated in different lairage pens in a German commercial pig abattoir. Samples of the water and the drinkers in different locations were microbiologically cultivated and examined. After new drinking equipment had been installed for one month, three months and five years, biofilm formation was detectable, and retrograde growth from the nipple drinkers was seen up to the connection with the main water distribution system. In particular, Enterobacteriaceae and Pseudomonas spp. were found in all samplings of the nipple drinkers. Zoonotic pathogens, Salmonella, pathogenic Yersinia enterocolitica and methicillin-resistant Staphylococcus aureus, were also isolated from the nipple drinkers, while Listeria monocytogenes was not detected via microbial cultivation methods in any of the samples. Since the pigs take the contaminated nipple drinkers into their mouths to drink, or drink contaminated water containing the pathogens, transmission and even infection of the pigs in the lairage can be assumed. This could consequently lead to contamination or cross-contamination of the meat during slaughter and processing and to a public health risk.

Study of a combined biological product with antibacterial and probiotic activity

Scientific relevance. The issue of antimicrobial resistance has acquired global significance, urging the search for and use of alternative antibacterial agents to treat infectious diseases. In particular, this situation prompts wider use of biotechnology-derived medicinal products based on bacteriophages and probiotics, including those of the metabolite type. The bacteriotropic (antimicrobial and probiotic) properties of pharmaceutical compositions based on bacteriophages and metabolites of probiotic bacteria suggest a qualitatively new combined antibacterial effect, as well as effectiveness against microorganisms resistant to antibiotics.Aim. The authors aimed to study the properties of a combined biological product with antibacterial and probiotic effects.Materials and methods. The study focused on a combined biological product consisting of a probiotic agent based on Lactobacillus exometabolites and a complex bacteriophage. The antimicrobial activity evaluation involved the Appelmans method, a disc diffusion method using lawn cultures homologous (Staphylococcus spp., Pseudomonas aeruginosa, Escherichia coli, Proteus spp., Enterococcus spp., Salmonella spp., and Shigella spp.) and non-homologous (Acinetobacter baumannii, Klebsiella pneumoniae, and Yersinia enterocolitica) to the complex bacteriophage, and a bioluminescence inhibition test using the genetically modified indicator strain E. coli lum+. The study used bacterial strains isolated from various clinical samples and biotopes in bacteriological laboratories of healthcare institutions in the Perm Territory in 2019–2022. The probiotic activity was assessed by the stimulating effect on model microorganisms of the normal flora.Results. The Appelmans method showed that the combined biological product had high antimicrobial activity against microorganisms homologous to the complex bacteriophage. The titres calculated for the combined biological product and its complex bacteriophage component were comparable and amounted to 10–6.6±0.01 and 10–6.9±0.01, respectively. The disc diffusion method demonstrated that the combined biological product had a more pronounced antibacterial effect on all tested strains than its individual components. The optical density values obtained with the combined biological product were 1.5 and 2 times higher than the values observed with control samples in Bifidobacterium bifidum 1 and Lactobacillus plantarum 8P-А3 cultures, respectively, which demonstrated the stimulating effect of the product on the normal flora. The study results suggest the compatibility of the phage and probiotic components of the combined biological product, as well as its high antimicrobial activity.Conclusions. The novel combined biological product has high specific and non-specific antimicrobial activity, which consists in the inhibition of pathogenic bacteria growth without affecting the normal flora. The combined biological product broadens the range of medicinal products having probiotic and antibacterial effects, particularly on microorganisms resistant to antibiotics.

Comparison of Multiple-Locus Variable-Number Tandem Repeat Analysis Profiles of Enteropathogenic Yersinia spp. Obtained from Humans, Domestic Pigs, Wild Boars, Rodents, Pork and Dog Food.

The enteropathogenic Yersinia genus is commonly detected in wildlife including wild boars. Difficulties in its cultivation may hamper subsequent epidemiological studies and outbreak investigations. Multiple-locus variable-number tandem repeat analysis (MLVA) of Yersinia (Y.) enterocolitica and Y. pseudotuberculosis has proven useful in source attribution and epidemiological studies but has hitherto relied on the analysis of isolates. In the present study, MLVA profiles generated from 254 isolates of Y. enterocolitica indicated similarities between human, pig and rodent isolates. Further, MLVA analyses of 13 Y. pseudotuberculosis pure-cultured isolates were compared to MLVA analyses performed directly on the 14 PCR-positive enrichment broths from which the isolates originated, which showed matching MLVA profiles. This indicates that MLVA analysis performed directly on enrichment broths could be a useful method for molecular epidemiological investigations. In addition, 10 out of 32 samples of wild boar minced meat obtained from private hunters and from approved wild-game-handling establishments were PCR-positive for the presence of Y. enterocolitica and may indicate a risk for public health.

The Novel Yersinia enterocolitica Telomere Phage vB_YenS_P840 Is Closely Related to PY54, but Reveals Some Striking Differences.

Telomere phages are a small group of temperate phages, whose prophages replicate as a linear plasmid with covalently closed ends. They have been isolated from some Enterobacteriaceae and from bacterial species living in aquatic environments. Phage PY54 was the first Yersinia (Y.) enterocolitica telomere phage isolated from a nonpathogenic O:5 strain, but recently a second telomeric Yersinia phage (vB_YenS_P840) was isolated from a tonsil of a wild boar in Germany. Both PY54 and vB_YenS_P840 (P840) have a siphoviridal morphology and a similar genome organization including the primary immunity region immB and telomere resolution site telRL. However, whereas PY54 only possesses one prophage repressor for the lysogenic cycle, vB_YenS_P840 encodes two. The telRL region of this phage was shown to be processed by the PY54 protelomerase under in vivo conditions, but unlike with PY54, a flanking inverted repeat was not required for processing. A further substantial difference between the phages is their host specificity. While PY54 infects Y. enterocolitica strains belonging to the serotypes O:5 and O:5,27, vB_YenS_P840 exclusively lyses O:3 strains. As the tail fiber and tail fiber assembly proteins of the phages differ significantly, we introduced the corresponding genes of vB_YenS_P840 by transposon mutagenesis into the PY54 genome and isolated several mutants that were able to infect both serotypes, O:5,27 and O:3.

Parasites and zoonotic bacteria in the feces of cats and dogs from animal shelters in Carinthia, Austria

Due to their close associations with humans, dogs and cats can be important reservoirs for zoonotic pathogens. In the current study 200 fecal samples of dogs (n = 70 samples) and cats (n = 130 samples) from animal shelters in Carinthia, southern Austria, were examined for the presence of parasites (fecal flotation and larval migration assay) and selected bacteria. Overall, 17.1% of the canine and 38.5% of the feline samples were positive for parasites (p < 0.001), most commonly Giardia duodenalis (dogs and cats), including potentially zoonotic genotypes revealed by multilocus genotyping, and Toxocara cati (cats). Cryptosporidium (C. felis), Cystoisospora spp. (dogs and cats), hookworms (dog), Trichuris (dog) Capillaria hepatica (cats), taeniids (cat), and Aelurostrongylus abstrusus (cat) were also found. Zoonotic bacteria were detected in 10.5% of the samples, Salmonella enterica (dogs), Campylobacter jejuni (dogs and cats) and Yersinia enterocolitica (cat) and were significantly associated with parasite infections in cats but not in dogs. Samples that were positive for several pathogens were common; especially G. duodenalis and T. cati were frequently found in association with each other, other parasites or bacteria. The spectrum of detected pathogens is comparable to that of other dog and cat populations in central Europe. However, since animals from shelters are frequently rehomed, diagnostic measures, appropriate hygiene and therapy as well as training of shelter staff are recommended to prevent zoonotic transmission of enteropathogens to staff or new owners. The presence of heteroxenic parasites, i.e. Aelurostrongylus abstrusus and Taenia taeniaeformis, and spurious excretion of Ca. hepatica in cats, indicates that these animals preyed on intermediate hosts, and that biosafety measures in pet shelters need to be evaluated for their efficacy in the prevention of pathogen transmission.

A lateral flow immunochromatographic strip based on RPA and quantum dot nanobeads for rapid identification of pathogenic Yersinia enterocolitica

To detect pathogenic Yersinia enterocolitica in meat, a lateral flow immunochromatographic strip targeting on virulence gene ail was developed, which was based on recombinase polymerase amplification (RPA) and quantum dot nanobeads (QDNBs). Under optimal conditions, the minimum detection limit of the strip was 9.6 × 103 CFU/mL, with a standard curve equation of y = 0.144x - 0.5098 (R2 = 0.9863), and the detection times including the RPA reaction at hand temperature (35–36.5 °C), could be controlled in 10 min. When testing spiked sample, the recoveries (%) of Yersinia enterocolitica at 9.6 × 104 CFU/mL were 44.27 ± 2.08 and 45.31 ± 3.12 in beef and pork samples and was 107.55 ± 1.30 in chicken samples. When testing 15 actual samples, only one pork sample was tested positive, consistent with polymerase chain reaction (PCR) results. The strips are body temperature-triggered and rapidly visualized, suitable for rapid field detection of pathogenic Yersinia enterocolitica.

Virulence Genotyping Profiles of Cefazolin Resistance Yersinia enterocolitica Isolated From Milk and Milk Products in Egypt

Virulence Genotyping Profiles of Cefazolin Resistance Yersinia enterocolitica Isolated From Milk and Milk Products in Egypt

Immune checkpoint inhibitor-induced neurotoxicity is not associated with seroprevalence of neurotropic infections

BackgroundImmune checkpoint inhibitors (ICI) substantially improve outcome for patients with cancer. However, the majority of patients develops immune-related adverse events (irAEs), which can be persistent and significantly reduce quality of life. Neurological irAEs occur in 1–5% of patients and can induce severe, permanent sequelae or even be fatal. In order to improve the diagnosis and treatment of neurological irAEs and to better understand their pathogenesis, we assessed whether previous neurotropic infections are associated with neurological irAEs.MethodsNeurotropic infections that might predispose to ICI-induced neurological irAEs were analyzed in 61 melanoma patients from 3 countries, the Netherlands, Australia and Germany, including 24 patients with neurotoxicity and 37 control patients. In total, 14 viral, 6 bacterial, and 1 protozoal infections previously reported to trigger neurological pathologies were assessed using routine serology testing. The Dutch and Australian cohorts (NL) included pre-treatment plasma samples of patients treated with neoadjuvant ICI therapy (OpACIN-neo and PRADO trials; NCT02977052). In the Dutch/Australian cohort a total of 11 patients with neurological irAEs were compared to 27 control patients (patients without neurological irAEs). The German cohort (LMU) consisted of serum samples of 13 patients with neurological irAE and 10 control patients without any documented irAE under ICI therapy.ResultsThe association of neurological irAEs with 21 possible preceding infections was assessed by measuring specific antibodies against investigated agents. The seroprevalence of all the tested viral (cytomegalovirus, Epstein-Barr-Virus, varicella-zoster virus, measles, rubella, influenza A and B, human herpes virus 6 and 7, herpes simplex virus 1 and 2, parvovirus B19, hepatitis A and E and human T-lymphotropic virus type 1 and 2), bacterial (Borrelia burgdorferi sensu lato, Campylobacter jejuni, Mycoplasma pneumoniae, Coxiella burnetti, Helicobacter pylori, Yersinia enterocolitica and Y. pseudotuberculosis) and protozoal (Toxoplasma gondii) infections was similar for patients who developed neurological irAEs as compared to control patients. Thus, the analysis provided no evidence for an association of described agents tested for seroprevalence with ICI induced neurotoxicity.ConclusionPrevious viral, bacterial and protozoal neurotropic infections appear not to be associated with the development of neurological irAEs in melanoma patients who underwent therapy with ICI across 3 countries. Further efforts are needed to unravel the factors underlying neurological irAEs in order to identify risk factors for these toxicities, especially with the increasing use of ICI in earlier stage disease.