Absence of Host-Specific Genes in Canine and Human Staphylococcus pseudintermedius as Inferred from Comparative Genomics

Staphylococcus pseudintermedius is an important pathogen in dogs that occasionally causes infections in humans as an opportunistic pathogen of elderly and immunocompromised people. This study compared the genomic relatedness and antimicrobial resistance genes using genome-wide association study (GWAS) to examine host association of canine and human S. pseudintermedius isolates. Canine (n = 25) and human (n = 32) methicillin-susceptible S. pseudintermedius (MSSP) isolates showed a high level of genetic diversity with an overrepresentation of clonal complex CC241 in human isolates. This clonal complex was associated with carriage of a plasmid containing a bacteriocin with cytotoxic properties, a CRISPR-cas domain and a pRE25-like mobile element containing five antimicrobial resistance genes. Multi-drug resistance (MDR) was predicted in 13 (41%) of human isolates and 14 (56%) of canine isolates. CC241 represented 54% of predicted MDR isolates from humans and 21% of predicted MDR canine isolates. While it had previously been suggested that certain host-specific genes were present the current GWAS analysis did not identify any genes that were significantly associated with human or canine isolates. In conclusion, this is the first genomic study showing that MSSP is genetically diverse in both hosts and that multidrug resistance is important in dog and human-associated S. pseudintermedius isolates.

Download Full-text

atpD gene sequencing, multidrug resistance traits, virulence-determinants, and antimicrobial resistance genes of emerging XDR and MDR-Proteus mirabilis

Scientific Reports ◽

10.1038/s41598-021-88861-w ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Abdelazeem M. Algammal ◽

Hany R. Hashem ◽

Khyreyah J. Alfifi ◽

Helal F. Hetta ◽

Norhan S. Sheraba ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Resistance Genes ◽

Proteus Mirabilis ◽

Biofilm Formation ◽

Gene Sequencing ◽

Opportunistic Pathogen ◽

Severe Illness ◽

Virulence Determinants ◽

Antimicrobial Resistance Genes ◽

Port Said

AbstractProteus mirabilis is a common opportunistic pathogen causing severe illness in humans and animals. To determine the prevalence, antibiogram, biofilm-formation, screening of virulence, and antimicrobial resistance genes in P. mirabilis isolates from ducks; 240 samples were obtained from apparently healthy and diseased ducks from private farms in Port-Said Province, Egypt. The collected samples were examined bacteriologically, and then the recovered isolates were tested for atpD gene sequencing, antimicrobial susceptibility, biofilm-formation, PCR detection of virulence, and antimicrobial resistance genes. The prevalence of P. mirabilis in the examined samples was 14.6% (35/240). The identification of the recovered isolates was confirmed by the atpD gene sequencing, where the tested isolates shared a common ancestor. Besides, 94.3% of P. mirabilis isolates were biofilm producers. The recovered isolates were resistant to penicillins, sulfonamides, β-Lactam-β-lactamase-inhibitor-combinations, tetracyclines, cephalosporins, macrolides, and quinolones. Using PCR, the retrieved strains harbored atpD, ureC, rsbA, and zapA virulence genes with a prevalence of 100%, 100%, 94.3%, and 91.4%, respectively. Moreover, 31.4% (11/35) of the recovered strains were XDR to 8 antimicrobial classes that harbored blaTEM, blaOXA-1, blaCTX-M, tetA, and sul1 genes. Besides, 22.8% (8/35) of the tested strains were MDR to 3 antimicrobial classes and possessed blaTEM, tetA, and sul1genes. Furthermore, 17.1% (6/35) of the tested strains were MDR to 7 antimicrobial classes and harbored blaTEM, blaOXA-1, blaCTX-M, tetA, and sul1 genes. Alarmingly, three strains were carbapenem-resistant that exhibited PDR to all the tested 10 antimicrobial classes and shared blaTEM, blaOXA-1, blaCTX-M, tetA, and sul1 genes. Of them, two strains harbored the blaNDM-1 gene, and one strain carried the blaKPC gene. In brief, to the best of our knowledge, this is the first study demonstrating the emergence of XDR and MDR-P.mirabilis in ducks. Norfloxacin exhibited promising antibacterial activity against the recovered XDR and MDR-P. mirabilis. The emergence of PDR, XDR, and MDR-strains constitutes a threat alarm that indicates the complicated treatment of the infections caused by these superbugs.

Download Full-text

Molecular epidemiology and antimicrobial resistance features of Acinetobacter baumannii clinical isolates from Pakistan

Annals of Clinical Microbiology and Antimicrobials ◽

10.1186/s12941-019-0344-7 ◽

2020 ◽

Vol 19 (1) ◽

Cited By ~ 2

Author(s):

Nabil Karah ◽

Fizza Khalid ◽

Sun Nyunt Wai ◽

Bernt Eric Uhlin ◽

Irfan Ahmad

Keyword(s):

Antimicrobial Resistance ◽

Molecular Epidemiology ◽

Acinetobacter Baumannii ◽

Resistance Genes ◽

Antimicrobial Agents ◽

Opportunistic Pathogen ◽

Clinical Isolates ◽

Diffusion Method ◽

Agar Disc ◽

Antimicrobial Resistance Genes

Abstract Background Acinetobacter baumannii is a Gram-negative opportunistic pathogen with a notorious reputation of being resistant to antimicrobial agents. The capability of A. baumannii to persist and disseminate between healthcare settings has raised a major concern worldwide. Methods Our study investigated the antibiotic resistance features and molecular epidemiology of 52 clinical isolates of A. baumannii collected in Pakistan between 2013 and 2015. Antimicrobial susceptibility patterns were determined by the agar disc diffusion method. Comparative sequence analyses of the ampC and blaOXA-51-like alleles were used to assign the isolates into clusters. The whole genomes of 25 representative isolates were sequenced using the MiSeq Desktop Sequencer. Free online applications were used to determine the phylogeny of genomic sequences, retrieve the multilocus sequence types (ST), and detect acquired antimicrobial resistance genes. Results Overall, the isolates were grouped into 7 clusters and 3 sporadic isolates. The largest cluster, Ab-Pak-cluster-1 (blaOXA-66 and ISAba1-ampC-19) included 24 isolates, belonged to ST2 and International clone (IC) II, and was distributed between two geographical far-off cities, Lahore and Peshawar. Ab-Pak-clusters-2 (blaOXA-66, ISAba1-ampC-2), and -3 (blaOXA-66, ISAba1-ampC-20) and the individual isolate Ab-Pak-Lah-01 (ISAba1-blaOXA-66, ISAba1-ampC-2) were also assigned to ST2 and IC II. On the other hand, Ab-Pak-clusters-4 (blaOXA-69, ampC-1), -5 (blaOXA-69, ISAba1-ampC-78), and -6A (blaOXA-371, ISAba1-ampC-3) belonged to ST1, while Ab-Pak-cluster-6B (blaOXA-371, ISAba1-ampC-8) belonged to ST1106, with both ST1 and ST1106 being members of IC I. Five isolates belonged to Ab-Pak-cluster-7 (blaOXA-65, ampC-43). This cluster corresponded to ST158, showed a well-delineated position on the genomic phylogenetic tree, and was equipped with several antimicrobial resistance genes including blaOXA-23 and blaGES-11. Conclusions Our study detected the occurrence of 7 clusters of A. baumannii in Pakistan. Altogether, 6/7 of the clusters and 45/52 (86.5%) of the isolates belonged to IC I (n = 9) or II (n = 36), making Pakistan no exception to the global domination of these two clones. The onset of ST158 in Pakistan marked a geographical dispersal of this clone beyond the Middle East and brought up the need for a detailed characterization.

Download Full-text

High antimicrobial resistance and plasmid-carrying resistance genes in swine-origin mcr-1-positive Escherichia coli in Guangxi, South China

10.21203/rs.2.13699/v1 ◽

2019 ◽

Author(s):

Jingzhi Yuan ◽

Xiaoye Wang ◽

Dali Shi ◽

Qiang Ge ◽

Xingxing Song ◽

...

Keyword(s):

Escherichia Coli ◽

Drug Resistance ◽

Antimicrobial Resistance ◽

South China ◽

Resistance Genes ◽

High Rate ◽

Multi Drug Resistance ◽

Drug Resistant ◽

E Coli ◽

Antimicrobial Resistance Genes

Abstract Background The discovery of mcr-1-positive Escherichia coli (MCRPEC), a notable superbug, attracted great attention worldwide. Swine-origin multi-drug resistance MCRPEC is a potential threat to public health and safety. To date, few detailed studies regarding swine-origin MCRPEC in Guangxi, South China, have been reported. Results In this study, thirty-three MCRPEC harbored mcr-1 genes were identified from 142 E. coli strains isolated from swine droppings and entrails in Guangxi in 2018. All MCRPEC isolates were assigned to 8 unique STs, including ST10, ST224 and ST410, which overlapped with the human-origin MCRPEC. Additionally, a total of six plasmid replicon types (IncFI, IncHI1, IncY, IncN, IncI1 and IncX1) were found. Moreover, the drug susceptibility of the MCRPEC isolates was tested with 27 antimicrobial agents belonging to 17 antimicrobial categories that are usually used in hospitals. There were 19 extended spectrum beta lactamase (ESBL) E. coli and 12 carbapenem resistant E. coli among the 33 MCRPEC strains. Importantly, the MCRPEC showed a high rate of resistance against two broad-spectrum carbapenem antibiotics, imipenem and meropenem, which are forbidden in livestock production use. Three MCRPEC strains were further identified to be extensively drug-resistant (XDR), and other isolates were recognized as multi-drug-resistant (MDR). Meanwhile, to detect whether plasmid-carrying antimicrobial resistance genes coexisted with the mcr-1 gene in the MCRPEC isolates, a total of 22 plasmid-carrying antimicrobial resistance genes were tested for. The results showed that four ESBL genes and one pAmpC gene were identified. Eight of the MCRPEC isolates also contained the carbapenem gene blaNDM-5, which could cause untreatable infections. Moreover, ten non-lactamase genes were also detected. Conclusion This study indicated that swine-origin MCRPEC isolated in Guangxi seemed to have a high rate of resistance to both regular and final line of defense drugs as well as drug resistance genes, which pose a great threat to human public safety and health.

Download Full-text

Methicillin-Resistant Staphylococcus aureus Clonal Complex 398 as a Major MRSA Lineage in Dogs and Cats in Thailand

Antibiotics ◽

10.3390/antibiotics10030243 ◽

2021 ◽

Vol 10 (3) ◽

pp. 243

Author(s):

Surawit Chueahiran ◽

Jitrapa Yindee ◽

Pongthai Boonkham ◽

Nipattra Suanpairintr ◽

Pattrarat Chanchaithong

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Resistance ◽

Resistance Genes ◽

Methicillin Resistant Staphylococcus Aureus ◽

Clonal Complex ◽

Clinical Samples ◽

Methicillin Resistant ◽

Class A ◽

Antimicrobial Resistance Genes ◽

Mrsa St398

The aim of this study was to present molecular and antimicrobial resistance characteristics of methicillin-resistant Staphylococcus aureus (MRSA) clonal complex (CC) 398 isolated from diseased dogs and cats in Thailand. A total of 20 MRSA isolates of 134 Staphylococcus aureus isolated from canine and feline clinical samples during 2017–2020 were CC398, consisting of sequence type (ST) 398 (18 isolates), ST5926 (1 isolate), and ST6563 (1 isolate) by multilocus sequence typing. spa t034 and staphylococcal cassette chromosome mec (SCCmec) V were predominantly associated with ST398. Intraclonal differentiation was present by additional spa (t1255, t4653), non-detectable spa, composite SCCmec with a hybrid of ccrA1B1+ccrC and class A mec complex, and DNA fingerprints by pulsed-field gel electrophoresis. The isolates essentially carried antimicrobial resistance genes, mediating multiple resistance to β-lactams (mecA, blaZ), tetracyclines [tet(M)], aminoglycosides [aac(6′)-Ie-aph(2′)-Ia], and trimethoprim (dfr). Livestock-associated MRSA ST398 resistance genes including lnu(B), lsa(E), spw, fexA, and tet(L) were heterogeneously found and lost in subpopulation, with the absence or presence of additional erm(A), erm(B), and ileS2 genes that corresponded to resistance phenotypes. As only a single CC398 was detected with the presence of intraclonal variation, CC398 seems to be the successful MRSA clone colonizing in small animals as a pet-associated MRSA in Thailand.

Download Full-text

The Pheno- and Genotypic Characterization of Porcine Escherichia coli Isolates

Microorganisms ◽

10.3390/microorganisms9081676 ◽

2021 ◽

Vol 9 (8) ◽

pp. 1676

Author(s):

Tanja Bernreiter-Hofer ◽

Lukas Schwarz ◽

Elke Müller ◽

Adriana Cabal-Rosel ◽

Maciej Korus ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Resistance Genes ◽

Multidrug Resistant ◽

Whole Genome Sequence ◽

Health Concern ◽

Multi Drug Resistance ◽

Virulence Determinants ◽

Minimal Inhibitory Concentrations ◽

E Coli ◽

Antimicrobial Resistance Genes

Escherichia (E.) coli is the main causative pathogen of neonatal and post-weaning diarrhea and edema disease in swine production. There is a significant health concern due to an increasing number of human infections associated with food and/or environmental-borne pathogenic and multidrug-resistant E. coli worldwide. Monitoring the presence of pathogenic and antimicrobial-resistant E. coli isolates is essential for sustainable disease management in livestock and human medicine. A total of 102 E. coli isolates of diseased pigs were characterized by antimicrobial and biocide susceptibility testing. Antimicrobial resistance genes, including mobile colistin resistance genes, were analyzed by PCR and DNA sequencing. The quinolone resistance-determining regions of gyrA and parC in ciprofloxacin-resistant isolates were analyzed. Clonal relatedness was investigated by two-locus sequence typing (CH clonotyping). Phylotyping was performed by the Clermont multiplex PCR method. Virulence determinants were analyzed by customized DNA-based microarray technology developed in this study for fast and economic molecular multiplex typing. Thirty-five isolates were selected for whole-genome sequence-based analysis. Most isolates were resistant to ampicillin and tetracycline. Twenty-one isolates displayed an ESBL phenotype and one isolate an AmpC β-lactamase-producing phenotype. Three isolates had elevated colistin minimal inhibitory concentrations and carried the mcr-1 gene. Thirty-seven isolates displayed a multi-drug resistance phenotype. The most predominant β-lactamase gene classes were blaTEM-1 (56%) and blaCTX-M-1 (13.71%). Mutations in QRDR were observed in 14 ciprofloxacin-resistant isolates. CH clonotyping divided all isolates into 51 CH clonotypes. The majority of isolates belonged to phylogroup A. Sixty-four isolates could be assigned to defined pathotypes wherefrom UPEC was predominant. WGS revealed that the most predominant sequence type was ST100, followed by ST10. ST131 was detected twice in our analysis. This study highlights the importance of monitoring antimicrobial resistance and virulence properties of porcine E. coli isolates. This can be achieved by applying reliable, fast, economic and easy to perform technologies such as DNA-based microarray typing. The presence of high-risk pathogenic multi-drug resistant zoonotic clones, as well as those that are resistant to critically important antibiotics for humans, can pose a risk to public health. Improved protocols may be developed in swine farms for preventing infections, as well as the maintenance and distribution of the causative isolates.

Download Full-text

Multidrug-Resistant Klebsiella Pneumoniae: A Retrospective Study in Manaus, Brazil

10.21203/rs.3.rs-1028514/v1 ◽

2021 ◽

Author(s):

Rafael Nakamura-Silva ◽

Louise Cerdeira ◽

Mariana Oliveira-Silva ◽

Karen Regina Carim da Costa ◽

Elder Sano ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Klebsiella Pneumoniae ◽

Resistance Genes ◽

Opportunistic Pathogen ◽

Multidrug Resistant ◽

Drug Resistant ◽

Extensively Drug Resistant ◽

Antimicrobial Resistance Genes ◽

Northern Brazil ◽

The City

Abstract Klebsiella pneumoniae is an opportunistic pathogen that can cause several infections, mainly in hospitalised or immunocompromised individuals. The spread of K. pneumoniae emerging virulent and multidrug-resistant clones is a worldwide concern and its identification is crucial to control these strains especially in hospitals. This article reports data related to multi-resistant K. pneumoniae strains, isolated from inpatients in the city of Manaus, Brazil, harbouring virulence and antimicrobial resistance genes, including high-risk international clones belonging to clonal group (CG) 258. Twenty-one strains isolated from different patients admitted to four hospitals in the city of Manaus, located in the state of Amazonas, Northern Brazil (Amazon Rainforest region) were evaluated. The majority of strains (61.9 % n = 13) were classified as multidrug-resistant (MDR), and five strains (23.8 %) as extensively drug-resistant (XDR). Several virulence and antimicrobial resistance genes were found among the strains and eight strains (38.1 %) presented the hypermucoviscous phenotype. MLST analysis demonstrated a great diversity of STs among the strains, totaling 12 different STs (ST11, ST23, ST198, ST277, ST307, ST340, ST378, ST462, ST502, ST3991, ST3993 and ST5209). Four of these (ST11, ST23, ST307 and ST340) belong to CG258.

Download Full-text

Culture-Independent Genotyping, Virulence and Antimicrobial Resistance Gene Identification of Staphylococcus aureus from Orthopaedic Implant-Associated Infections

Microorganisms ◽

10.3390/microorganisms9040707 ◽

2021 ◽

Vol 9 (4) ◽

pp. 707

Author(s):

J. Christopher Noone ◽

Fabienne Antunes Ferreira ◽

Hege Vangstein Aamot

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Resistance ◽

Resistance Genes ◽

Virulence Gene ◽

Orthopaedic Implant ◽

Metagenomic Sequencing ◽

Gene Detection ◽

Shotgun Metagenomic Sequencing ◽

Antimicrobial Resistance Genes ◽

Culture Independent

Our culture-independent nanopore shotgun metagenomic sequencing protocol on biopsies has the potential for same-day diagnostics of orthopaedic implant-associated infections (OIAI). As OIAI are frequently caused by Staphylococcus aureus, we included S. aureus genotyping and virulence gene detection to exploit the protocol to its fullest. The aim was to evaluate S. aureus genotyping, virulence and antimicrobial resistance genes detection using the shotgun metagenomic sequencing protocol. This proof of concept study included six patients with S. aureus-associated OIAI at Akershus University Hospital, Norway. Five tissue biopsies from each patient were divided in two: (1) conventional microbiological diagnostics and genotyping, and whole genome sequencing (WGS) of S. aureus isolates; (2) shotgun metagenomic sequencing of DNA from the biopsies. Consensus sequences were analysed using spaTyper, MLST, VirulenceFinder, and ResFinder from the Center for Genomic Epidemiology (CGE). MLST was also compared using krocus. All spa-types, one CGE and four krocus MLST results matched Sanger sequencing results. Virulence gene detection matched between WGS and shotgun metagenomic sequencing. ResFinder results corresponded to resistance phenotype. S. aureus spa-typing, and identification of virulence and antimicrobial resistance genes are possible using our shotgun metagenomics protocol. MLST requires further optimization. The protocol has potential application to other species and infection types.

Download Full-text