Molecular Detection of Antibiotic-Resistant Genes in Pseudomonas aeruginosa from Nonclinical Environment: Public Health Implications in Mthatha, Eastern Cape Province, South Africa

Evaluation of resistant profiles and detection of antimicrobial-resistant genes of bacterial pathogens in the nonclinical milieu is imperative to assess the probable risk of dissemination of resistant genes in the environment. This paper sought to identify antibiotic-resistant genes in Pseudomonas aeruginosa from nonclinical sources in Mthatha, Eastern Cape, and evaluate its public health implications. Samples collected from abattoir wastewater and aquatic environment were processed by membrane filtration and cultured on CHROMagarTM Pseudomonas medium. Species identification was performed by autoSCAN-4 (Dade Behring Inc., IL). Molecular characterization of the isolates was confirmed using real-time polymerase chain reaction (rPCR) and selected isolates were further screened for the possibility of harboring antimicrobial resistance genes. Fifty-one Pseudomonas species were recovered from abattoir wastewater and surface water samples, out of which thirty-six strains were Pseudomonas aeruginosa (70.6%). The P. aeruginosa isolates demonstrated resistance to aztreonam (86.1%), ceftazidime (63.9%), piperacillin (58.3%), cefepime (55.6%), imipenem (50%), piperacillin/tazobactam (47.2%), meropenem (41.7%), and levofloxacin (30.6%). Twenty out of thirty-six P. aeruginosa displayed multidrug resistance profiles and were classified as multidrug-resistant (MDR) (55.6%). Most of the bacterial isolates exhibited a high Multiple Antibiotic Resistance (MAR) Index ranging from 0.08 to 0.69 with a mean MAR index of 0.38. In the rPCR analysis of fifteen P. aeruginosa isolates, 14 isolates (93.3%) were detected harboring blaSHV, six isolates (40%) harbored blaTEM, and three isolates (20%) harbored blaCTX-M, being the least occurring ESBL. Results of the current study revealed that P. aeruginosa isolates recovered from nonclinical milieu are resistant to frontline clinically relevant antipseudomonal drugs. This is concerning as it poses a risk to the environment and constitutes a public health threat. Given the public health relevance, the paper recommends monitoring of multidrug-resistant pathogens in effluent environments.

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Antibiotic resistant gene of Pseudomonas aeruginosa from non-clinical environment: public health implications in Mthatha, Eastern Cape Province, South Africa

10.21203/rs.3.rs-31330/v1 ◽

2020 ◽

Author(s):

Mojisola C. Hosu ◽

Sandeep Vasaikar ◽

Grace. E. Okuthe ◽

Teke Apalata

Keyword(s):

Public Health ◽

Pseudomonas Aeruginosa ◽

Antibiotic Resistance ◽

Multidrug Resistant ◽

Eastern Cape ◽

Clinical Environment ◽

Antibiotic Resistant ◽

Health Implications ◽

Clinical Environments ◽

Mar Index

Abstract Background Pseudomonas aeruginosa (P. aeruginosa) are Gram-negative bacilli that are ubiquitous in nature particularly plentiful in soils and in aquatic milieu because they thrive well in humid or wet environments. It has extensive metabolic versatility, ability to thrive well and colonize diverse ecological habitats such as soil, rhizosphere, wastewaters that, enhances its potential threat to public health. Abattoirs, aquatic ecosystem including generated wastewaters are latent sources of pathogenic bacteria, serve as reservoirs and contribute to the spread of antibiotic resistant genes. Several studies have focused on clinical environment while scarce data exist from non-clinical environments, which sometimes are hotspots of antibiotic resistance. Thus, the present investigation aimed to identify antibiotic resistant gene of P. aeruginosa from non-clinical sources in Mthatha, Eastern Cape and evaluate its public health implications. Results Fifty-five (55) Pseudomonas species and other organisms recovered from effluent and surface water samples included P. aeruginosa (65.4%), P. fluorescens (27.3%), Burkholderia gladioli (5.5%) and Burkholderia cepacia (1.8%). The P. aeruginosa isolates showed high resistance to aztreonam (86.1%), ceftazidime (63.9%), piperacillin (58.3%) and cefepime (55.6%); with moderate resistance displayed to imipenem (50%), piperacillin/tazobactam (47.2%), meropenem (41.7%) and levofloxacin (30.6%). Twenty out of thirty-six (36) P. aeruginosa displayed multidrug resistance profiles and were classified as multidrug resistant (MDR) (55.6%). Most of the bacterial isolates exhibited a high Multiple Antibiotic Resistance (MAR) Index of > 0.2 ranging from 0.08–0.69 with a mean MAR Index of 0.38. PCR analysis of fifteen (15) P. aeruginosa isolates detected 14 (93.3%) harbored blaSHV, 6 (40%) harbored blaTEM with the least occurring ESBL as blaCTX−M at 3 (20%). Conclusions Results of the current study indicating resistance of environmental P. aeruginosa isolates to front-line clinically relevant antipseudomonal drugs is concerning and poses risk to the environment and receiving water bodies. Given the public health relevance, the results of this study highlight the importance and urgent need of monitoring multidrug-resistant pathogens in effluent environments. These non-clinical environments are potential reservoirs of resistance genes that would further serve as avenues for the dissemination of multidrug resistant bacteria in the community.

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Ecological and Public Health Implications of the Discharge of Multidrug-Resistant Bacteria and Physicochemical Contaminants from Treated Wastewater Effluents in the Eastern Cape, South Africa

Water ◽

10.3390/w9080562 ◽

2017 ◽

Vol 9 (8) ◽

pp. 562 ◽

Cited By ~ 5

Author(s):

Martins Adefisoye ◽

Anthony Okoh

Keyword(s):

Public Health ◽

South Africa ◽

Multidrug Resistant ◽

Treated Wastewater ◽

Resistant Bacteria ◽

Eastern Cape ◽

Multidrug Resistant Bacteria ◽

Health Implications ◽

Wastewater Effluents

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The Prevalence of Exoenzyme S Gene in Multidrug-Sensitive and Multidrug-Resistant Pseudomonas aeruginosa Clinical Strains

Polish Journal of Microbiology ◽

10.5604/01.3001.0010.6500 ◽

2017 ◽

Vol 66 (4) ◽

pp. 427-431 ◽

Cited By ~ 1

Author(s):

Tomasz Bogiel ◽

Aleksander Deptuła ◽

Joanna Kwiecińska-Piróg ◽

Małgorzata Prażyńska ◽

Agnieszka Mikucka ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Virulence Factors ◽

Multidrug Resistant ◽

Hospital Environment ◽

Antibiotic Resistant ◽

Exoenzyme S ◽

Clinical Specimens ◽

Antimicrobial Resistance Genes ◽

The Difference ◽

Susceptibility Patterns

Pseudomonas aeruginosa rods are one of the most commonly isolated microorganisms from clinical specimens, usually responsible for nosocomial infections. Antibiotic-resistant P. aeruginosa strains may present reduced expression of virulence factors. This fact may be caused by appropriate genome management to adapt to changing conditions of the hospital environment. Virulence factors genes may be replaced by those crucial to survive, like antimicrobial resistance genes. The aim of this study was to evaluate, using PCR, the occurrence of exoenzyme S-coding gene (exoS) in two distinct groups of P. aeruginosa strains: 83 multidrug-sensitive (MDS) and 65 multidrug-resistant (MDR) isolates. ExoS gene was noted in 72 (48.7%) of the examined strains: 44 (53.0%) MDS and 28 (43.1%) MDR. The observed differences were not statistically significant (p = 0.1505). P. aeruginosa strains virulence is rather determined by the expression regulation of the possessed genes than the difference in genes frequency amongst strains with different antimicrobial susceptibility patterns.

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Detection of extended spectrum beta-lactamase genes in Pseudomonas aeruginosa isolated from patients in rural Eastern Cape Province, South Africa

Scientific Reports ◽

10.1038/s41598-021-86570-y ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Mojisola C. Hosu ◽

Sandeep D. Vasaikar ◽

Grace E. Okuthe ◽

Teke Apalata

Keyword(s):

Pseudomonas Aeruginosa ◽

Antimicrobial Resistance ◽

Multidrug Resistant ◽

High Rate ◽

Infection Prevention And Control ◽

Beta Lactamase ◽

Eastern Cape ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum ◽

Resistance Patterns

AbstractThe proliferation of extended spectrum beta-lactamase (ESBL) producing Pseudomonas aeruginosa represent a major public health threat. In this study, we evaluated the antimicrobial resistance patterns of P. aeruginosa strains and characterized the ESBLs and Metallo- β-lactamases (MBL) produced. Strains of P. aeruginosa cultured from patients who attended Nelson Mandela Academic Hospital and other clinics in the four district municipalities of the Eastern Cape between August 2017 and May 2019 were identified; antimicrobial susceptibility testing was carried out against thirteen clinically relevant antibiotics using the BioMérieux VITEK 2 and confirmed by Beckman autoSCAN-4 System. Real-time PCR was done using Roche Light Cycler 2.0 to detect the presence of ESBLs; blaSHV, blaTEM and blaCTX-M genes; and MBLs; blaIMP, blaVIM. Strains of P. aeruginosa demonstrated resistance to wide-ranging clinically relevant antibiotics including piperacillin (64.2%), followed by aztreonam (57.8%), cefepime (51.5%), ceftazidime (51.0%), piperacillin/tazobactam (50.5%), and imipenem (46.6%). A total of 75 (36.8%) multidrug-resistant (MDR) strains were observed of the total pool of isolates. The blaTEM, blaSHV and blaCTX-M was detected in 79.3%, 69.5% and 31.7% isolates (n = 82), respectively. The blaIMP was detected in 1.25% while no blaVIM was detected in any of the strains tested. The study showed a high rate of MDR P. aeruginosa in our setting. The vast majority of these resistant strains carried blaTEM and blaSHV genes. Continuous monitoring of antimicrobial resistance and strict compliance towards infection prevention and control practices are the best defence against spread of MDR P. aeruginosa.

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Serotype distribution, antimicrobial susceptibility, antimicrobial resistance genes and virulence genes of Salmonella isolated from a pig slaughterhouse in Yangzhou, China

AMB Express ◽

10.1186/s13568-019-0936-9 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 2

Author(s):

Quan Li ◽

Jian Yin ◽

Zheng Li ◽

Zewei Li ◽

Yuanzhao Du ◽

...

Keyword(s):

Public Health ◽

Antimicrobial Resistance ◽

Resistance Genes ◽

Antimicrobial Susceptibility ◽

Virulence Genes ◽

Multidrug Resistant ◽

Resistance Rate ◽

Economic Losses ◽

Production Chain ◽

Antimicrobial Resistance Genes

AbstractSalmonella is an important food-borne pathogen associated with public health and high economic losses. To investigate the prevalence and the characteristics of Salmonella in a pig slaughterhouse in Yangzhou, a total of 80 Salmonella isolates were isolated from 459 (17.43%) samples in 2016–2017. S. Derby (35/80, 43.75%) was the most prevalent, followed by S. Rissen (16/80, 20.00%) and S. Newlands (11/80, 13.75%). The highest rates of susceptibility were observed to cefoxitin (80/80, 100.0%) and amikacin (80/80, 100.0%), followed by aztreonam (79/80, 98.75%) and nitrofurantoin (79/80, 98.75%). The highest resistance rate was detected for tetracycline (65/80, 81.25%), followed by ampicillin (60/80, 75.00%), bactrim (55/80, 68.75%), and sulfisoxazole (54/80, 67.50%). Overall, 91.25% (73/80) of the isolates were resistant to at least one antibiotic, while 71.25% (57/80) of the isolate strains were multidrug resistant in the antimicrobial susceptibility tested. In addition, 86.36% (19/22) of the 22 antimicrobial resistance genes in the isolates were identified. Our data indicated that the resistance to certain antimicrobials was significantly associated, in part, with antimicrobial resistance genes. Furthermore, 81.25% (65/80) isolates harbored the virulence gene of mogA, of which 2 Salmonella Typhimurium isolates carried the mogA, spvB and spvC virulence genes at the same time. The results showed that swine products in the slaughterhouse were contaminated with multidrug resistant Salmonella commonly, especially some isolates carry the spv virulence genes. The virulence genes might facilitate the dissemination of the resistance genes to consumers along the production chain, suggesting the importance of controlling Salmonella during slaughter for public health.

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MULTIPLE ANTIBIOTIC RESISTANCE INDEXING OF NON- FERMENTING GRAM NEGATIVE BACILLI

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2017.v10i6.17717 ◽

2017 ◽

Vol 10 (6) ◽

pp. 78

Author(s):

Bhuvaneshwari Gunasekar

Keyword(s):

Antibiotic Resistance ◽

Tertiary Care ◽

Multidrug Resistant ◽

Diffusion Method ◽

Care Hospital ◽

Multiple Antibiotic Resistance ◽

Antibiotic Resistant ◽

Gram Negative ◽

Mar Index ◽

Surgery Department

Objective: The multiple antibiotic resistance (MAR) indexing and finding Multidrug resistant (MDR) bacteria will help to indicate the origin from high risk of contamination where the antibiotics are often used. Hence this study was carried out to give the MAR index of non-fermenting Gram negative bacilli in a tertiary care hospital which would help our infection control team also.Methods: Drug resistance was tested by Kirby bauer’s disc diffusion method. MAR index was calculated using the formula, a/b (were a= number of antibiotics to which the organism was resistant and b= total number of antibiotics to which the organism was tested).Results: Out of 240 Gram negative non-fermenters isolated, 117 (49%) strains were greater than 0.2 of MAR index, 95(81%) was from in-patient department. 73(62%) were hospitalized for more than 3 days, 44 (38%) was from surgery department. 49(42%) was wound specimen. Out of 117 multiple antibiotic resistant isolates 99 (85%) were MDR isolates.Conclusion: 51% prevalence of isolates >0.2 MAR index shows that the source of contamination can still be brought up down by proper surveillance and management with proper usage of surface and skin disinfectants especially in surgery ward where the MAR index has indicated more usage of antibiotics

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F-type Pyocins are Diverse Non-Contractile Phage Tail-Like Weapons for Killing Pseudomonas aeruginosa

10.1101/2021.02.16.431561 ◽

2021 ◽

Author(s):

Senjuti Saha ◽

Chidozie D. Ojobor ◽

Erik Mackinnon ◽

Olesia I. North ◽

Joseph Bondy-Denomy ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Bactericidal Activity ◽

Pathogenic Bacteria ◽

Therapeutic Potential ◽

Experimental Studies ◽

Opportunistic Pathogen ◽

Multidrug Resistant ◽

Antibiotic Resistant ◽

Genes Encoding ◽

Intraspecies Competition

ABSTRACTMost Pseudomonas aeruginosa strains produce bacteriocins derived from contractile or non-contractile phage tails known as R-type and F-type pyocins, respectively. These bacteriocins possess strain-specific bactericidal activity against P. aeruginosa and likely increase evolutionary fitness through intraspecies competition. R-type pyocins have been studied extensively and show promise as alternatives to antibiotics. Although they have similar therapeutic potential, experimental studies on F-type pyocins are limited. Here, we provide a bioinformatic and experimental investigation of F-type pyocins. We introduce a systematic naming scheme for genes found in R- and F-type pyocin operons and identify 15 genes invariably found in strains producing F-type pyocins. Five proteins encoded at the 3’-end of the F-type pyocin cluster are divergent in sequence, and likely determine bactericidal specificity. We use sequence similarities among these proteins to define 11 distinct F-type pyocin groups, five of which had not been previously described. The five genes encoding the variable proteins associate in two modules that have clearly re-assorted independently during the evolution of these operons. These proteins are considerably more diverse than the specificity-determining tail fibers of R-type pyocins, suggesting that F-type pyocins emerged earlier or have been subject to distinct evolutionary pressures. Experimental studies on six F-type pyocin groups show that each displays a distinct spectrum of bactericidal activity. This activity is strongly influenced by the lipopolysaccharide O-antigen type, but other factors also play a role. F-type pyocins appear to kill as efficiently as R-type pyocins. These studies set the stage for the development of F-type pyocins as anti-bacterial therapeutics.IMPORTANCEPseudomonas aeruginosa is an opportunistic pathogen that causes a broad spectrum of antibiotic resistant infections with high mortality rates, particularly in immunocompromised individuals and cystic fibrosis patients. Due to the increasing frequency of multidrug-resistant P. aeruginosa infections, there is great interest in the development of alternative therapeutics. One alternative is protein-based antimicrobials called bacteriocins, which are produced by one strain of bacteria to kill other strains. In this study, we investigate F-type pyocins, bacteriocins naturally produced by P. aeruginosa that resemble non-contractile phage tails. We show that they are potent killers of P. aeruginosa, and distinct pyocin groups display different killing specificities. We have identified the probable specificity determinants of F-type pyocins, which opens up the potential to engineer them to precisely target strains of pathogenic bacteria. The resemblance of F-type pyocins to well characterized phage tails will greatly facilitate their development into effective antibacterials.

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First identification and Limited Dissemination of mcr-1 Colistin Resistance in Salmonella Isolates in Jiaxing

Journal of Food Protection ◽

10.4315/jfp-21-069 ◽

2021 ◽

Author(s):

Ping Li ◽

Li Zhan ◽

Henghui Wang ◽

Wenjie Gao ◽

Lei Gao ◽

...

Keyword(s):

Public Health ◽

Multidrug Resistance ◽

Antimicrobial Resistance ◽

Gastrointestinal Disease ◽

Foodborne Pathogen ◽

Multidrug Resistant ◽

Colistin Resistance ◽

Antimicrobial Resistance Genes ◽

Genetic Features ◽

Multidrug Resistant Strain

Salmonella , a major foodborne pathogen, causes severe gastrointestinal disease in people and animals worldwide. Plasmid-borne mcr-1 , which confers colistin resistance in Salmonella, has significant epidemiological interest for public health safety. Here, we report the first evidence of mcr-1 -mediated colistin resistance in one multidrug-resistant strain，namely 16062 in this study, from 355 Salmonella isolates collected for Jiaxing foodborne pathogen monitoring in Zhejiang Province in 2015–2019. In addition to colistin, 16062 displayed multidrug resistance to various antimicrobials (β-lactams, quinolone, sulfonamide, florfenicol, ampicillin, streptomycin, nalidixic acid, aminoglycoside, and trimethoprim-sulfamethox). The mcr-1 -carrying IncX4 plasmid (p16062-MCR) in this study shares a conserved structure with other mcr -IncX4 plasmids. We found that other antimicrobial-resistance genes ( aac(6')-Ib-cr , aadA1 , aadA2 , aph(3')-Ia , oqxA , oqxB , sul1 , and cmlA1 ) are located on p16062-cmlA, an atypical IncHI2 plasmid, in isolate 16062. This is the first identification of transferable colistin resistance in foodborne Salmonella isolate collected in Jiaxing city, the 5-year monitoring of which revealed limited dissemination. By determining the genetic features of the plasmid vehicle, the characteristics of transferable mcr genes circulating in isolates from Jiaxing are now clearer.

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Antibiotics in Food Chain: The Consequences for Antibiotic Resistance

Antibiotics ◽

10.3390/antibiotics9100688 ◽

2020 ◽

Vol 9 (10) ◽

pp. 688

Author(s):

Shashi B. Kumar ◽

Shanvanth R. Arnipalli ◽

Ouliana Ziouzenkova

Keyword(s):

Public Health ◽

Antibiotic Resistance ◽

Food Chain ◽

Bacterial Infections ◽

Antibiotic Resistance Genes ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Alternative Strategies ◽

Antibiotic Resistant ◽

Continuous Use

Antibiotics have been used as essential therapeutics for nearly 100 years and, increasingly, as a preventive agent in the agricultural and animal industry. Continuous use and misuse of antibiotics have provoked the development of antibiotic resistant bacteria that progressively increased mortality from multidrug-resistant bacterial infections, thereby posing a tremendous threat to public health. The goal of our review is to advance the understanding of mechanisms of dissemination and the development of antibiotic resistance genes in the context of nutrition and related clinical, agricultural, veterinary, and environmental settings. We conclude with an overview of alternative strategies, including probiotics, essential oils, vaccines, and antibodies, as primary or adjunct preventive antimicrobial measures or therapies against multidrug-resistant bacterial infections. The solution for antibiotic resistance will require comprehensive and incessant efforts of policymakers in agriculture along with the development of alternative therapeutics by experts in diverse fields of microbiology, biochemistry, clinical research, genetic, and computational engineering.

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Role of biofilm-specific antibiotic resistance genes PA0756-0757, PA5033 and PA2070 in Pseudomonas aeruginosa

10.1101/341826 ◽

2018 ◽

Author(s):

Prasanth Manohar ◽

Thamaraiselvan Shanthini ◽

Reethu Ann Philip ◽

Subramani Ramkumar ◽

Manali Kale ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Antibiotic Resistance ◽

Resistance Genes ◽

Biofilm Formation ◽

Tamil Nadu ◽

Antibiotic Resistance Genes ◽

Clinical Samples ◽

Cross Sectional ◽

Antibiotic Resistant ◽

Resistant Genes

AbstractTo evaluate the presence of biofilm-specific antibiotic-resistant genes, PA0756-0757, PA5033 and PA2070 in Pseudomonas aeruginosa isolated from clinical samples in Tamil Nadu. For this cross-sectional study, 24 clinical isolates (included pus, urine, wound, and blood) were collected from two diagnostic centers in Chennai from May 2015 to February 2016. Biofilm formation was assessed using microtiter dish biofilm formation assay and minimal inhibitory concentration (MIC) and minimal bactericidal concentrations (MBC) were determined for planktonic and biofilm cells (MBC assay). Further, PCR amplification of biofilm-specific antibiotic resistance genes PA0756-0757, PA5033 and PA2070 were performed. Biofilm formation was found to be moderate/strong in 16 strains. MBC for planktonic cells showed that 4, 7, 10 and 14 strains were susceptible to gentamicin, ciprofloxacin, meropenem and colistin respectively. In MBC assay for biofilm cells (MBC-B), all the 16 biofilm producing strains were resistant to ciprofloxacin and gentamicin whereas nine and four were resistant to meropenem, and colistin respectively. The biofilm-specific antibiotic-resistant genes PA0756-0757 was found in 10 strains, 6 strains with PA5033 and 9 strains with PA2070 that were found to be resistant phenotypically. This study highlighted the importance of biofilm-specific antibiotic resistance genes PA0756-0757, PA5033, and PA2070 in biofilm-forming P. aeruginosa.

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