Tox Gene Research Articles

Anti-quorum sensing and anti-biofilm activities of Pasteurella multocida strains

A total of 52 Pasteurella multocida strains of capsular serogroups (A, B and D) were screened for anti-quorum sensing activity against Chromobacterium violaceum. Of which, 12 strains of serogroups A were found to possess anti-quorum sensing activity. Inhibition activity was highest for strain NIVEDIPm9 and lowest for strain NIVEDIPm30 based on zone of pigment inhibition. Further, cell free extract of NIVEDIPm9 strain showed highest anti-biofilm activity in reference E. coli strain and concentration dependent degradation activity of C6-AHL molecule. In whole genome sequence annotation of NIVEDIPm9 strain predicted the presence of four metallo-β-lactamases (MBL) fold metallo-hydrolase proteins. In docking studies, MBL1 and MBL3 proteins showed high binding affinity with autoinduce signalling molecules AHL compound of OH-C10, binding energy value were −6.3 and −6.2 kcal/mol. Interaction study of VAF and quorum sensing molecules showed that OmpA and HgbA proteins were stimulated by all the ten molecules (C4-AHLs, C6-AHLs, C10-AHLs, C14-AHLs, 3-oxo-C10-AHLs, 3OH-C10-HSL, C8-HSL, C10-HSL, C12-HSL, C14-HSL), while toxA gene was stimulated by OH-C10-AHL molecule, sodC gene was stimulated by none. In conclusion, we described the anti-quorum sensing activities of diverse P. multocida strains causing Pasteurellosis in livestock.

Porcine Nose Atrophy Assessed by Automatic Imaging and Detection of Bordetella bronchiseptica and Other Respiratory Pathogens in Lung and Nose

The nasal mucosa is a crucial filtering organ to prevent attachment and invasion of pathogens. To assess nasal health in relation to lung health, transverse cross sections of the nasal turbinates of 121 pigs suffering from respiratory disease and sent for diagnostic necropsy were scored visually and by an artificial intelligence (AI) medical diagnostic application (AI DIAGNOS), resulting in a high correlation of both scores (p < 0.001). Nasal samples of the diseased pigs were examined only for Bordetella (B.) bronchiseptica (PCR and bacteriological culture) and Pasteurella (P.) multocida (bacteriological culture). All pigs showed various degrees of inflammatory lung tissue alterations, and 35.5% of the pigs had atrophy of the nasal turbinates with no relation to detection rates of B. bronchiseptica (54.5%) and P. multocida (29.0%) in the nose. All P. multocida strains from nose samples were negative for the toxA gene so non-progressive atrophic rhinitis was diagnosed. Pigs positive for B. bronchiseptica in the nose were more often positive for B. bronchiseptica in the lung (p < 0.001) and for other bacterial species in the lower respiratory tract (p = 0.005). The new diagnostic application for scoring cross sections of nasal turbinates is a valuable tool for a fast and reproducible diagnostic.

Genetic profiling of virulence genes among Pseudomonas Aeruginosa isolates recovered from diseased chickens

Pseudomonas aeruginosa (P. aeruginosa) is regarded as an opportunistic organism for bird species. Also is one can be mentioned as an example of an environment-associated infection that results in severe disease in poultry farms, Indeed, we detected the incidence of P. aeruginosa in chickens and also discovered the implicated virulence genes and antibiotic-resistant mechanism. Two hundred cases were collected from broiler and layer chicken organs (heart blood, heart, and lungs) that suffered from some respiratory manifestation and septicemia in addition to yolk sacs from one-day-old chicks, which were examined bacteriologically and Serologically. The studied isolates were tested against eight antimicrobial discs. Bacterial DNA was analyzed using polymerase chain reaction (PCR) against six virulence genes (lasL, lasB toxA, exoU, exoT, and oprl genes). Eighteen isolates out of 200 (9%) of P. aeruginosa were defined. Serologically the isolates were related to more serogroup (O1,O4,O6, O11 and O12) P. aeruginosa. PCR defined the existence of P. aeruginosa by detection of the 16SRNA gene. All isolates showed 100% multidrug resistance (MDR) to more than one antimicrobial disc (sulfamethoxazole, erythromycin, tetracycline, and ampicillin). Conversely, those isolates exhibited 100% sensitivity to ciprofloxacin, and four of them showed sensitivity to norfloxacin with a percentage of (22.2%). In addition to 15 isolates showed resistance of 83.3% percent to streptomycin and nalidixic acid. By PCR test, all examined P. aeruginosa strains carried four virulence genes (lasI, toxA, exoU, and oprl). However, only three strains carried lasB, and four carried the exoT gene. In conclusion, we confirmed the presence of different factors of virulence for P. aeruginosa and MDR to more than one antimicrobial drug. So, restricting Pseudomonas infection in poultry requires strict biosecurity measures to control and eradicate the infection by P. aeruginosa

Resistance patterns, virulence determinants, and biofilm genes of multidrug-resistant Pseudomonas aeruginosa isolated from fish and fish handlers

Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic bacterium that is widely distributed in aquatic environments and causes major economic losses in fish and public health hazards.This study aimed to identify the occurrence of P. aeruginosa in samples collected from fish and fish handlers, and to investigate the antimicrobial susceptibility, virulence determinants, and biofilm genes of P. aeruginosa isolates. A total of 276 samples were cross-sectionally collected from Nile tilapia (53), Golden grey mullet (52), Mediterranean horse mackerel (50), Striped red mullet (71), and fish handlers (50) at five different retail fish markets in Damietta Governorate, Egypt. Pseudomonas species (spp.) were biochemically identified in 57.9% of the total examined samples. Peudomonas aeruginosa were the most prevalent species isolated from the fish and human samples via PCR technique. Peudomonas aeruginosa isolates exhibited full resistance (100%) to tobramycin (TOB), gentamicin (CN), and colistin (CL), with a high level of susceptibility (88.5%) to imipenem (IPM) using the disk diffusion method. Most P. aeruginosa isolates (84.6%) exhibited drug resistance, with 61.5% were multidrug resistance (MDR) and 23.1% were extensive drug resistance (XDR). Most isolates had at least four virulence-associated genes (lasB, toxA, exoU, and oprL) and three biofilm genes (psIA, peIA, and lasR) by using uniplex PCR. The lasI, and rhlR Quorum Sensing (QS) genes were identified in 84.6% and 61.5% in the examined P. aeruginosa isolates, respectively. The highest mortality rate in Nile tilapia experimentally infected with P. aeruginosa isolate encoding most of virulent genes. Multivariate analyses revealed high heterogeneity among the examined isolates. This study revealed the emergence of virulent and drug resistant P. aeruginosa isolates in fish, poses high risks to consumers and food. Thus, strict hygienic measures should be considered when catching, handling, and storing fish, in addition to the routine application of antimicrobial susceptibility testing.

Identification of Corynebacterium ulcerans and Erysipelothrix sp. in Malayan pangolins-a potential threat to public health?

This study firstly reports the presence of two potentially zoonotic bacteria, Corynebacterium ulcerans and Erysipelothrix sp., in diseased Malaysian pangolins collected in 2019. The pangolin C. ulcerans is lethal in mice and resists many antibiotics. It clustered with a lethal human strain but lacked the diphtheria toxin gene. Diphtheria toxin is widely used as a vaccine around the world to protect humans from the infection of corynebacteria. The lack of the tox gene suggests that the current vaccine may be of limited efficacy against this pangolin strain. The pangolin Erysipelothrix sp. is the sister clade of Erysipelothrix rhusiopathiae. It is lethal in mice, and two commercial vaccines failed to protect the mice against challenge with the pangolin Erysipelothrix sp., such that this strain could potentially pose a threat to the swine industry. These findings emphasize the potential threat of pangolin bacteria.

Impeding microbial biofilm formation and Pseudomonas aeruginosa virulence genes using biologically synthesized silver Carthamus nanoparticles

Long-term antibiotic treatment results in the increasing resistance of bacteria to antimicrobials drugs, so it is necessary to search for effective alternatives to prevent and treat pathogens that cause diseases. This study is aimed for biological synthesis of silver Carthamus nanoparticles (Ag-Carth-NPs) to combat microbial biofilm formation and Pseudomonas aeruginosa virulence genes. Ag-Carth-NPs are synthesized using Carthamus tenuis aqueous extract as environmentally friendly method has no harmful effect on environment. General factorial design is used to optimize Ag-Carth-NPs synthesis using three variables in three levels are Carthamus extract concentration, silver nitrate concentration and gamma radiation doses. Analysis of response data indicates gamma radiation has a significant effect on Ag-Carth-NPs production. Ag-Carth-NPs have sharp peak at λ max 425 nm, small and spherical particles with size 20.0 ± 1.22 nm, high stability up to 240 day with zeta potential around − 43 ± 0.12 mV, face centered cubic crystalline structure and FT-IR spectroscopy shows peak around 620 cm−1 that corresponding to AgNPs that stabilized by C. tenuis extract functional moiety. The antibacterial activity of Ag-Carth-NPs against pathogenic bacteria and fungi was determined using well diffusion method. The MIC values of Ag-Carth-NPs were (6.25, 6.25, 3.126, 25, 12.5, 12.5, 25 and 12.5 µg/ml), MBC values were (12.5, 12.5, 6.25, 50, 25, 25, 50 and 25 µg/ml) and biofilm inhibition% were (62.12, 68.25, 90.12, 69.51, 70.61, 71.12, 75.51 and 77.71%) against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, Candida tropicalis and Candida albicans respectively. Ag-Carth-NPs has bactericidal efficacy and significantly reduced the swarming, swimming motility, pyocyanin and protease production of P. aeruginosa. Furthermore, P. aeruginosa ToxA gene expression was significantly down regulated by 81.5%, while exoU reduced by 78.1%, where lasR gene expression reduction was 68%, while the reduction in exoU was 66% and 60.1% decrease in lasB gene expression after treatment with Ag-Carth-NPs. This activity is attributed to effect of Ag-Carth-NPs on cell membrane integrity, down regulation of virulence gene expression, and induction of general and oxidative stress in P. aeruginosa. Ag-Carth-NPs have no significant cytotoxic effects on normal human cell (Hfb4) but have IC50 at 5.6µg/mL against of HepG-2 cells. Limitations of the study include studies with low risks of silver nanoparticles for in vitro antimicrobial effects and its toxicity.

Prevalence Of Some Pyogenic Organisms In Skin Wound Infections In Sheep And Goat

Background: Pyogenic bacterial infections are broadly found worldwide among sheep and goats and are adversely impacting the development of the small ruminants’ industry. The study was performed to identify and characterize most prevalent bacteria isolated from skin and wound infections of sheep and goat of various rearing condition in Egypt. Materials and Methods: One hundred and fifteen pus samples (sheep≈70) and (goat≈45) obtained from skin abscesses and pyogenic wounds during the study period 2022-2023. The samples were cultured onto suitable media, microscopically and biochemically tested. The obtained isolates were assessed for their antibiotic susceptibility and subjected to polymerase chain reaction to detect certain genes which responsible for the virulence and antibiotic resistance. Results: Bacterial isolates were recovered from 83 out of the 115 collected pus samples (72.17%) with pure isolation were obtained in 68 cultures. The bacterial isolation was achieved in 88.57% and 46.66% from sheep and goat respectively. Pseudomonas aeruginosa were recovered as 19.35% from sheep and 23.81% from goat while Staphylococcus aureus were isolated as 46.77% and 38.09% from sheep and goat respectively. Regarding Corynebacterium pseudotuberculosis was obtained from 37.09% of sheep and 9.52% of goat samples. P. aeruginosa isolates were highly sensitive to erythromycin, amikacin, ampicillin /sulbactam and cefoxitin and high resistance to amoxicillin /clavulanic acid, clindamycin, and quinolones with 58.82% MDR pattern. S. aureus isolates revealed the high sensetivity to amikacin, imipenem and clindamycin but high resistant to quinolones with 21.62% act as MDR. C. pseudotuberculosis isolates were sensitive to most of tested antibiotics except amoxicillin /clavulanic acid and clindamycin. PCR assays revealed that P. aeruginosa isolates carried toxA and blaOXA-1 genes by 35.29% and 64.70% respectively while S. aureus isolates was found to load eta gene by 13.51% and ermC gene 18.91% and in the same line ten C. pseudotuberculosis was found to harbor pld gene 40%. Conclusion: It was concluded that Pseudomonas aeruginosa, Staphylococcus aureus as well as Corynebacterium pseudotuberculosis were the most isolated bacterial species in skin infection among sheep and goat in Egypt. Majority of the isolates were MDR and highly resistant to β lactams and quinolones but sensitive to amikacin and imipenem. PCR assays revealed that the isolates carried virulence and antibiotic resistance genes in significant incidence.

Identification of effector genes of <i>Parastagonospora nodorum</i>, <i>P. pseudonodorum</i> in Tambov populations and sensitivity genes to NEs in varieties and hybrid lines of spring soft wheat

BACKGROUND: Parastagonospora nodorum and P. pseudonodorum are known for their ability to produce necrotrophic effectors that play an important role in micromycete pathogenicity. Fungal strains characterized by the presence of effector genes will be used to create artificial infectious backgrounds to identify sources and donors of resistance to fungal diseases. Selected varieties and lines that are donors of recessive alleles snn1 and snn3, which control plant resistance to fungal toxins PtrTox1 and PtrTox3, are recommended for inclusion in programs for breeding wheat for resistance to septoriosis pathogens. AIM: The aim of the work is to assess the populations of the fungi Parastagonospora nodorum and P. pseudonodorum in 2023 based on the presence of effector genes, as well as to identify alleles of Snn1/snn1, Snn3/snn3 genes that control the sensitivity or resistance of wheat to PtrTox1 and PtrTox3 toxins. MATERIALS AND METHODS: Infectious samples were collected in 2023 from spring wheat leaves. In addition, the material for the study was 2 varieties and 23 lines of spring soft wheat of local selection. Using the molecular markers Xfcp624 and Xcfd20, the presence of the Snn1 and Snn3-B1 alleles, which control sensitivity to the fungal toxins PtrTox1 and PtrTox3, was detected. RESULTS: Using molecular screening, ToxA and Tox1 genes were identified in genotypes of P. pseudonodorum isolates; Tox3 and Tox267 in P. nodorum isolates. 2 varieties of spring soft wheat and 11 hybrid lines carry a recessive allele snn1, which protects against the phytopathogen toxin PtrTox1; wheat variety Tambovchanka and 2 hybrid lines carry the recessive allele snn3 on chromosome B1, which confers resistance to the fungal toxin PtrTox3. CONCLUSIONS: The ToxA gene was found only among monoconidial isolates of P. pseudonodorum species obtained from leaves of spring soft wheat of Lebedushka variety. As a result of molecular screening, the Tox1 gene was identified among 70 P. pseudonodorum isolates (43,21% of those studied). The presence of the Tox3 and Tox267 genes was established in 30 isolates of P. nodorum species obtained from plant samples of spring durum wheat Donskaya Elegiya. Using the PCR method, donors of the snn1 and snn3 genes were identified.

Challenges of Diphtheria Toxin Detection.

Diphtheria toxin (DT) is the main virulence factor of Corynebacterium diphtheriae, C. ulcerans and C. pseudotuberculosis. Moreover, new Corynebacterium species with the potential to produce diphtheria toxin have also been described. Therefore, the detection of the toxin is the most important test in the microbiological diagnosis of diphtheria and other corynebacteria infections. Since the first demonstration in 1888 that DT is a major virulence factor of C. diphtheriae, responsible for the systemic manifestation of the disease, various methods for DT detection have been developed, but the diagnostic usefulness of most of them has not been confirmed on a sufficiently large group of samples. Despite substantial progress in the science and diagnostics of infectious diseases, the Elek test is still the basic recommended diagnostic test for DT detection. The challenge here is the poor availability of an antitoxin and declining experience even in reference laboratories due to the low prevalence of diphtheria in developed countries. However, recent and very promising assays have been developed with the potential for use as rapid point-of-care testing (POCT), such as ICS and LFIA for toxin detection, LAMP for tox gene detection, and biosensors for both.

First Pangenome of Corynebacterium rouxii, a Potentially Toxigenic Species of Corynebacterium diphtheriae Complex

Corynebacterium rouxii is one of the recently described species of the Corynebacterium diphtheriae complex. As this species can potentially infect different hosts and harbor the tox gene, producing diphtheria toxin, we present its first pangenomic analysis in this work. A total of fifteen genomes deposited in online databases were included. After confirming the taxonomic position of the isolates by genomic taxonomy, the genomes were submitted to genomic plasticity, gene synteny, and pangenome prediction analyses. In addition, virulence and antimicrobial resistance genes were investigated. Finally, epidemiological data were obtained through molecular typing, clustering, and phylogenetic analysis. Our data demonstrated genetic diversity within the species with low synteny. However, the gene content is extensively conserved, and the pangenome is composed of 2606 gene families, of which 1916 are in the core genome and 80 are related to unique genes. Prophages, insertion sequences, and genomic islands were found. A type I-E CRISPR-Cas system was also detected. Besides the tox gene, determinants involved in adhesion and iron acquisition and two putative antimicrobial resistance genes were predicted. These findings provide valuable insight about this species’ pathogenicity, evolution, and diversity. In the future, our data can contribute to different areas, including vaccinology and epidemiology.

The Virulence Genes of Sensitive Strain Pseudomonas aeruginosa Causing Nosocomial Outbreaks

Pseudomonas aeruginosa is an opportunistic pathogen which is commonly associated with healthcare associated infection. They possess multiple pathogenic factors which play a role in causing invasive infections such as surgical site infection, pneumonia, and blood stream infection. There were two hospital outbreaks caused by sensitive strains P. aeruginosa between 2016 and 2017 involving 17 patients. The outbreak investigation by Pulsed Field Gel Electrophoresis (PFGE) revealed seven clonally related P. aeruginosa strains (A-G). This study aims to determine the virulence factors acquired by the P. aeruginosa isolates and describe the clinical outcome of the patients. Seventeen P. aeruginosa isolates from the stocked collection were retrieved for six virulence genes, namely ToxA, ExoS, LasI, LasB, OprI, and OprL by PCR. Ten out of 17 of the P. aeruginosa isolates were able to revive. The ExoS, LasI, LasB, OprI, and OprL genes, respectively, were detected in all isolates, while ToxA gene was detected in six isolates which belonged to clone A (one isolate) and clone C (five isolates). The isolate from clone A caused pneumonia and isolates from clone C caused surgical site infections which led to disseminated infections and death. The presence of multiple virulence genes in these P. aeruginosa isolates may have contributed to the invasiveness, and the outcome of the infection. More studies with a larger number of patients will give a better insight regarding the actual role of these genes in different clinical manifestations caused by sensitive strain P. aeruginosa.

Evaluation of PA0756 and toxA Gene Expression in Clinical Isolates of Pseudomonas aeruginosa and Its Comparison Between Biofilm-Producing and Planktonic Isolates

Background: Biofilm formation is crucial in the pathogenesis of Pseudomonas aeruginosa infections, contributing to increased antibiotic resistance. Objectives: This study aimed to evaluate the expression of the PA0756 and toxA genes in clinical isolates of P. aeruginosa, comparing biofilm-producing isolates with planktonic ones. Methods: In this cross-sectional study, 160 clinical isolates of P. aeruginosa were collected from hospitals in Tabriz. Biofilm development was assessed using a 96-well flat-bottom microtiter plate test, followed by polymerase chain reaction (PCR) amplification of the resistance genes PA0756 and toxA. DNA synthesis for both biofilm and planktonic P. aeruginosa was conducted, with the quantification of target genes carried out using real-time PCR with specific primers. Results: Of the isolates, 139 (87%) formed biofilms. These strains showed high resistance to gatifloxacin, piperacillin, gentamycin, and ceftazidime (82.25%, 72.97%, 76.12%, and 74.63%, respectively), while resistance to colistin and polymyxin B was low (2.5% and 3%, respectively). The expression study revealed significant upregulation of the PA0756 and toxA genes in biofilm formers by 1.2 - 5.6 fold and 1.2 - 2.5 fold, respectively, compared to planktonic cells (P = 0.012 and P = 0.041, respectively). Conclusions: The PA0756 and toxA genes are significantly involved in biofilm-specific antibiotic resistance in P. aeruginosa, presenting potential targets for therapeutic interventions against antibiotic resistance caused by this pathogen.

Molecular characterization of Pasteurella multocida from cats and antibiotic sensitivity of the isolates.

Companion animals, including dogs and cats, are frequently identified as sources of Pasteurella multocida, a bacterium that can be transmitted to humans and cause infections. This survey defines the prevalence, antibiotic sensitivity, capsular types, lipopolysaccharide (LPS) types and virulence factors of P. multocida isolated from cats. A total of 100 specimens from various cat breeds were collected. P. multocida was characterized using both biochemical tests and PCR. Genotypes of isolates were determined using capsular and LPS typing methods. Additionally, virulotyping was performed by detecting the presence of 12 virulence-associated genes. Disk diffusion was used to determine the antibiotic sensitivity of the isolates. The prevalence of P. multocida in cats was 29%. Among the isolates, the majority were capsular type A (96.5%) and type D (3.4%), with a predominant presence of type A. Twenty-six of the isolates (89.66%) belonged to LPS genotype L6, whereas three isolates (10.3%) belonged to genotype L3. Among the 12 virulence genes examined, sodC, oma87, ptfA, nanB and ompH showed remarkable prevalence (100%). The toxA gene was detected in four isolates (13.8%). Variations were observed in other virulence genes. The nanH gene was present in 93.1% of the isolates, whereas the pfhA gene was detected in 58.6% of the isolates. The exbD-tonB, hgbB, sodA and hgbA genes showed prevalence rates of 96.5%, 96.5%, 96.5% and 82.8%, respectively. Additionally, particular capsule and LPS types were associated with specific virulence genes. Specifically, the toxA and pfhA genes were found to be more prevalent in isolates with capsular type A and LPS genotype L6. Most isolates were resistant to ampicillin, clindamycin, lincomycin, streptomycin and penicillin. According to this epidemiological and molecular data, P. multocida from cats possess several virulence-associated genes and are resistant to antimicrobial medicines commonly used in humans and animals. Thus, it is crucial to consider the public health concerns of P. multocida in humans.

Molecular Identification and Detection of Virulence Genes Among Pseudomonas Aeruginosa Isolated from Burns Infections

Objective: Virulence factors are substances produced by pathogenic Pseudomonas aeruginosa that contribute significantly to the etiology of disease. These virulence factors are encoded by virulence genes found on the Pseudomonas aeruginosa chromosome. Methods: Between July 2021 and June 2022, 71 Pseudomonas aeruginosa isolates were identified from burn wounds at the Burn and Plastic Surgery Hospital in Duhok, Iraq. The lasB and toxA genes were identified using Polymerase Chain Reaction (PCR). Results: Only 26.36% (29/71) of the 71 Pseudomonas aeruginosa isolates were found in males, whereas 38.18% (42/71) were found in females. Furthermore, 76.06% (54/71) of the isolates were multidrug resistant. They demonstrated greater resistance to piperacillin, 98.59% resistance rates. Among the isolates analyzed, 35 (64.81%) were positive for toxA and 27 (50%) were positive for lasB genes. Conclusion: Due to the limited number of effective medications against this bacteria that are currently available, all isolates must undergo antimicrobial susceptibility testing. By doing this, you can help manage the treatment plan and stop the emergence of resistance in burn units.

Draft genome sequence of novel Candidatus Ornithobacterium hominis carrying antimicrobial resistance genes in Egypt

BackgroundCandidatus Ornithobacterium hominis (O. hominis), which was identified in nasopharyngeal swabs from Egypt, has been associated with respiratory disorders in humans. O. hominis, a recently identified member of the Flavobacteriaceae family, belongs to the largest family within the Bacteroidetes phylum. This family includes hundreds of species and 90 genera, including major human pathogens such as Capnocytophaga canimorsus and Elizabethkingia meningoseptica. Herein, we presented two draft genome assemblies of O. hominis that were extracted from metagenomic data using the Illumina sequencing method. The alignment of reads against the O. hominis genome was accomplished using BLASTN, and the reads with significant hits were extracted using Seqtk and assembled using SPAdes. The primary goal of this study was to obtain a more profound understanding of the genomic landscape of O. hominis, with an emphasis on identifying the associated virulence, antimicrobial genes, and distinct defense mechanisms to shed light on the potential role of O. hominis in human respiratory infections.ResultsThe genome size was estimated to be 1.84 Mb, including 1,931,660 base pairs (bp), with 1,837 predicted coding regions and a G+C content of 35.62%. Genes encoding gliding motility, antibiotic resistance (20 genes), and the toxA gene were all included in the genome assembly. Gliding motility lipoproteins (GldD, GldJ, GldN, and GldH) and the gliding motility-associated ABC transporter substrate-binding protein, which acts as a crucial virulence mechanism in Flavobacterium species, were identified. The genome contained unique genes encoding proteins, such as the ParE1 toxin that defend against the actions of quinolone and other antibiotics. The cobalt-zinc-cadmium resistance gene encoding the protein CzcB, which is necessary for metal resistance, urease regulation, and colonization, was also detected. Several multidrug resistance genes encoding proteins were identified, such as MexB, MdtK, YheI, and VanC.ConclusionOur study focused on identifying virulence factors, and antimicrobial resistance genes present in the core genome of O. hominis. These findings provide valuable insights into the potential pathogenicity and antibiotic susceptibility of O. hominis.

Adipose-derived stem cell exosomes promote tumor characterization and immunosuppressive microenvironment in breast cancer

Adipose-derived stem cells (ASC) or autologous fat transplantation could be used to ameliorate breast cancer postoperative deformities. This study aims to explore the action of ASC and ASC-exosomes (ASC-exos) in breast cancer characterization and tumor microenvironment immunity, which provided a new method into the application of ASC-exos. ASC were extracted from human adipose tissue for the isolation and verification of ASC-exos. ASC-exos were co-cultured with CD4+T cells, CD14+ monocytes and MCF-7 cells, respectively. The tumor formation of nude mice was also constructed. Cell characterization was determined by CCK8, scratch assay, and Transwell. Hematoxylin–eosin (HE), immunohistochemistry (IHC) and immunofluorescence (IF) staining were used to observe the histopathology and protein expression. CD4+T cell and CD14+ monocytes differentiation was detected by flow cytometry. Western blot, qRT-PCR and RNAseq were used to detect the action of ASC-exos on gene and protein expression. CD4+T cells could take up ASC-exos. ASC-exos inhibited Th1 and Th17 differentiation and promoted Treg differentiation of CD4+T cells. ASC-exos inhibited M1 differentiation and promoted M2 differentiation of CD14+ monocytes. ASC-exos promoted the migration, proliferation, and invasion, while inhibited apoptosis of MCF-7 cells. ASC-exos promoted the tumor formation of breast cancer. The effect of ASC-exos on tumor microenvironment immunity was in accordance with the above in vitro results. TOX, CD4 and LYZ1 genes were upregulated, while Mettl7b and Serpinb2 genes were downregulated in ASC-exos group. Human T-cell leukemia virus 1 infection pathway was significantly enriched in ASC-exos. Thus, ASC-exos promoted breast cancer characterization and tumor microenvironment immunosuppression by regulating macrophage and T cell differentiation.

Dissemination of mcr-1 and β-lactamase genes among Pseudomonas aeruginosa: molecular characterization of MDR strains in broiler chicks and dead-in-shell chicks infections

ObjectivesPseudomonas aeruginosa (P. aeruginosa) is one of the most serious pathogens implicated in antimicrobial resistance, and it has been identified as an ESKAPE along with other extremely significant multidrug resistance pathogens. The present study was carried out to explore prevalence, antibiotic susceptibility phenotypes, virulence-associated genes, integron (int1), colistin (mcr-1), and β-lactamase resistance' genes (ESBls), as well as biofilm profiling of P. aeruginosa isolated from broiler chicks and dead in-shell chicks.DesignA total of 300 samples from broiler chicks (n = 200) and dead in-shell chicks (n = 100) collected from different farms and hatcheries located at Mansoura, Dakahlia Governorate, Egypt were included in this study. Bacteriological examination was performed by cultivation of the samples on the surface of both Cetrimide and MacConkey’s agar. Presumptive colonies were then subjected to biochemical tests and Polymerase Chain Reaction (PCR) targeting 16S rRNA. The recovered isolates were tested for the presence of three selected virulence-associated genes (lasB, toxA, and exoS). Furthermore, the retrieved isolates were subjected to phenotypic antimicrobial susceptibility testing by Kirby–Bauer disc diffusion method as well as phenotypic detection of ESBLs by both Double Disc Synergy Test (DDST) and the Phenotypic Confirmatory Disc Diffusion Test (PCDDT). P. aeruginosa isolates were then tested for the presence of antibiotic resistance genes (ARGs): int1, mcr-1, and ESBL genes (OXA-10, OXA-2, VEB-1, SHV, TEM, and CTX-M). Additionally, biofilm production was examined by the Tube Adherent method (TA) and Microtiter Plate assay (MTP).ResultsFifty –five isolates were confirmed to be P. aeruginosa, including 35 isolates from broiler chicks and 20 isolates from dead in-shell chicks. The three tested virulence genes (lasB, toxA, and exoS) were detected in all isolates. Antibiogram results showed complete resistance against penicillin, amoxicillin, ceftriaxone, ceftazidime, streptomycin, erythromycin, spectinomycin, and doxycycline, while a higher sensitivity was observed against meropenem, imipenem, colistin sulfate, ciprofloxacin, and gentamicin. ESBL production was confirmed in 12 (21.8%) and 15 (27.3%) isolates by DDST and PCDDT, respectively. Antibiotic resistance genes (ARGs): int1, mcr-1, and ESBL genes (OXA-10, SHV, TEM, and CTX-M), were detected in 87.3%, 18.2%, 16.4%, 69.1%, 72.7%, and 54.5% of the examined isolates respectively, whereas no isolate harbored the OXA-2 or VEB-1 genes. Based on the results of both methods used for detection of biofilm formation, Kappa statistics [kappa 0.324] revealed a poor agreement between both methods.Conclusionsthe emergence of mcr-1 and its coexistence with other resistance genes such as β-lactamase genes, particularly blaOXA-10, for the first time in P. aeruginosa from young broiler chicks and dead in-shell chicks in Egypt pose a risk not only to the poultry industry but also to public health.Graphical

Resistance of old winter bread wheat landraces to tan spot

Background. The most effective and environmentally safe way to combat wheat diseases is to produce cultivars resistant to their pathogens. For this purpose, old landraces are often used as genetically diverse sources of traits important for breeding. In the process of wheat breeding for resistance to tan spot caused by the fungus Pyrenophora tritici-repentis (Died.) Drechs. (abbr. Ptr), selection is carried out against the dominant allele of Tsn1, the gene of sensitivity to the toxin Ptr ToxA, which induces necrosis and represents the main pathogenicity factor of Ptr controlled by the ToxA gene. The aim of the study was to characterize a set of bread wheat (Triticum aestivum L.) accessions from the VIR collection for resistance to various Ptr populations, genotype these accessions using Xfcp623 – a DNA marker of the Tsn1 gene, and identify sources of tan spot resistance.Materials and methods. Sixty-seven accessions of winter bread wheat landraces were studied. Seedling resistance to two Ptr populations was assessed using a 5-point scale adopted at VIZR. The allelic state of Tsn1 was identified by PCR.Results. Dominant alleles of Tsn1 were found for 55% of the studied accessions. Seventeen accessions were resistant or moderately resistant to two Ptr populations and an isolate from Krasnodar Territory previously used for their characterization. Nine of them had the tsn1tsn1 genotype, and 8 had Tsn1Tsn1. The accessions mainly belonged to three agroecological groups proposed by N. I. Vavilov: “steppe winter bread wheat (Banatka wheats)”, “North European forest awnless bread wheats (Sandomirka wheats)”, and “Caucasian mountain winter bread wheat”.Conclusion. The identified 17 accessions resistant to Ptr are potential breeding sources of resistance. In the studied set of accessions, no significant relationship was found between the allelic state of the Tsn1 gene in the accession and its response to the infection with pathogen populations, including isolates with the ToxA gene.

Antibacterial effect of Moringa oleifera on Staphylococcus aureus and Pseudomonas aeruginosa isolated from raw milk and some dairy products with special reference to biofilm gene expression.

Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus) are well defined as food poisoning pathogens that are highly resistant and need continuous studies. The purpose of the work was to examine phenotypic and genotypic characteristics of both P. aeruginosa and S. aureus, and treatment trials with medicinal plants. Samples were examined for isolation of P. aeruginosa and S. aureus on selective media followed by biochemical confirmation, biofilm formation, genes detection, and expression of P. aeruginosa pslA biofilm gene was performed by quantitative real-time polymerase chain reaction after treatment with 0.312 mg/ml Moringa oleifera aqueous extract as a minimum inhibitory concentration. The highest isolation rate of P. aeruginosa was 20% from both raw milk and Kariesh cheese, followed by 16% and 12% from ice cream and processed cheese, respectively, while the highest isolation rate of S. aureus was 36% from raw milk followed by 28% in ice cream and 16% in both Kariesh cheese and processed cheese. 30% of P. aeruginosa isolates were biofilm producers, while only 21% of S. aureus isolates were able to produce biofilm. The P. aeruginosa isolates harbor virulence-associated genes nan1, exoS, toxA, and pslA at 100%, 80%, 40%, and 40%, respectively. Staphylococcus aureus SEs genes were examined in S. aureus strains, where SEA and SEB genes were detected with 60%, but no isolate harbored SEC, SED, or SEE. The significant fold change of P. aeruginosa pslA expression was 0.40332 after treatment with M. oleifera aqueous extract. Pseudomonas aeruginosa and S. aureus harbor dangerous virulence genes that cause food poisoning, but M. oleifera extract could minimize their action.

Proteomics of Toxigenic Corynebacteria.

Within the genus Corynebacterium, six species are potential carriers of the tox gene, which encodes the highly potent diphtheria exotoxin: Corynebacterium diphtheriae, Corynebacterium belfantii, Corynebacterium rouxii, Corynebacterium ulcerans, Corynebacterium pseudotuberculosis and Corynebacterium silvaticum. Based on their potential to infect different host species and cause either human infections, zoonotic diseases or infections of economically important animals, these bacteria are of high scientific and economic interest and different research groups have carried out proteome analyses. These showed that especially the combination of MS-based proteomics with bioinformatic tools helped significantly to elucidate the functional aspects of corynebacterial genomes and to handle the genome and proteome complexity. The combination of proteomic and bioinformatic approaches was also used to discover new vaccine and drug targets. In addition, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has been established as a fast and precise tool for the identification of these bacteria.