Characterisation of Vibrio Species from Surface and Drinking Water Sources and Assessment of Biocontrol Potentials of Their Bacteriophages

The aim of this study was to characterise Vibrio species of water samples collected from taps, boreholes, and dams in the North West province, South Africa, and assess biocontrol potentials of their bacteriophages. Fifty-seven putative Vibrio isolates were obtained on thiosulfate-citrate-bile-salt-sucrose agar and identified using biochemical tests and species-specific PCRs. Isolates were further characterised based on the presence of virulence factors, susceptibility to eleven antibiotics, and biofilm formation potentials. Twenty-two (38.60%) isolates were confirmed as Vibrio species, comprising V. harveyi (45.5%, n = 10), V. parahaemolyticus (22.7%, n = 5), V. cholerae (13.6%, n = 3), V. mimicus (9.1%, n = 2), and V. vulnificus (9.1%, n = 2). Three of the six virulent genes screened were positively amplified; four V. parahaemolyticus possessed the tdh (18.18%) and trh (18.18%) genes, while the zot gene was harboured by 3 V. cholerae (13.64%) and one V. mimicus (4.55%) isolate. Isolates revealed high levels of resistance to cephalothin (95.45%), ampicillin (77.27%), and streptomycin (40.91%), while lower resistances (4.55%–27.27%) were recorded for other antimicrobials. Sixteen (72.7%) isolates displayed multiple antibiotic-resistant properties. Cluster analysis of antibiotic resistance revealed a closer relationship between Vibrio isolates from different sampling sites. The Vibrio species displayed biofilm formation potentials at 37°C (63.6, n = 14), 35°C (50%, n = 11), and 25°C (36.4%, n = 8). Two phages isolated in this study (vB_VpM_SA3V and vB_VcM_SA3V) were classified as belonging to the family Myoviridae based on electron microscopy. These were able to lyse multidrug-resistant V. parahaemolyticus and V. cholerae strains. These findings not only indicate the presence of antibiotic-resistant virulent Vibrio species from dam, borehole, and tap water samples that could pose a health risk to humans who either come in contact with or consume water but also present these lytic phages as alternative agents that can be exploited for biological control of these pathogenic strains.

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Diversity, antimicrobial characterization and biofilm formation of Enterococci isolated from aquaculture and slaughterhouse sources in Benin City, Nigeria

Ife Journal of Science ◽

10.4314/ijs.v22i3.4 ◽

2021 ◽

Vol 22 (3) ◽

pp. 51-63

Author(s):

E.O. Igbinosa ◽

A. Beshiru ◽

E.E.O. Odjadjare

Keyword(s):

Water Samples ◽

Biofilm Formation ◽

Continuous Monitoring ◽

Enterococcus Species ◽

Bacterial Strains ◽

Antibiotic Resistant ◽

Benin City ◽

Primer Sets ◽

Species Specific ◽

Aquaculture Facilities

The present study was designed to characterize Enterococci isolates obtained from water samples at aquaculture and slaughterhouse facilities in Benin City, Nigeria. A total of 144 water samples were collected from aquaculture and slaughterhouse facilities. All samples were analyzed using classical microbiological and molecular-based methods. Enterococci were identified using specific primer sets (genus and species specific primers) and are as follows: E. faecalis 36 (25.5%); E. faecium 39 (27.7%); E. durans 19 (13.4%); E. casseliflavus 13 (9.2%); E. hirae 14 (9.9%) and other Enterococcus species 20 (14.2%). The resistance profile of the bacterial strains to antibiotics was as follows: [tetracycline (n=67, 47%)]; [vancomycin (n=74, 52%)]; [erythromycin (n=91, 64%)] and [penicillin (n=141, 100%)]. Enterococci virulence genes detected include: [gelE (n=120, 85.1%)]; [cylA (n= 52, 36.9%)]; [hyl (n=96, 68.1%)]; [esp (n=135, 95.8%)]; [ace (n= 127, 90.1%)] and [agg n=118, 83.7%)]. Antibiotic-resistant gene detected from the phenotypic resistant isolates were 55/74 (74.3%) vanA; 61/67 (91.1%) tetC; 122/141 (86.5%) blapse1 and 62/91 (68.1%) ermA. Antibiotic-resistant coupled with biofilm formation potential of Enterococcus species include penicillin+biofilm 116 (82.3%); erythromycin+biofilm 85 (60.3%); and vancomycin+biofilm 74 (52.3%). Findings from this study reveal that strains with the ability of forming biofilms have enhanced antimicrobial resistance. Continuous monitoring of slaughterhouses and aquaculture facilities is necessary to guarantee food safety. Key Words: Aquaculture, Biofilm,Enterococcus, Environments, Resistance, Slaughterhouse

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Rapid, specific and quantitative polymerase chain reaction (PCR) detection of pathogenic protozoa Entamoeba histolytica for drinking water supply

Water Science & Technology Water Supply ◽

10.2166/ws.2011.057 ◽

2011 ◽

Vol 11 (4) ◽

pp. 418-425 ◽

Cited By ~ 1

Author(s):

S. W. Lam ◽

H. B. Zhang ◽

L. Yu ◽

C. H. Woo ◽

K. N. Tiew ◽

...

Keyword(s):

Real Time ◽

Water Samples ◽

Real Time Pcr ◽

Genomic Dna ◽

Tap Water ◽

Pcr Detection ◽

Raw Water ◽

Conventional Pcr ◽

Polymerase Chain ◽

Species Specific

In this study, a quantitative species-specific polymerase chain reaction (PCR) method to rapidly detect E. histolytica in water is developed. First, the specificity of E. histolytica PCR detection was verified by using species-specific primers of 16S-like rRNA genes to clearly differentiate it from the closely related amoebae species E. dispar and E. moshkovskii. The sensitivity of this method was subsequently determined using purified E. histolytica genomic DNA and culture cells as PCR reaction templates. Results indicated that conventional PCR visualized on 1% agarose gel was able to detect as low as 0.02 pg genomic DNA and 5 cells, while real-time PCR could detect 0.01 pg genomic DNA and 2 cells of E. histolytica. The protocols for E. histolytica PCR detection in real water samples were then optimized by spiking E. histolytica cells into tap water and reservoir raw water samples. A two-round centrifugation treatment to concentrate amoeba cells directly as a PCR template was the most effective way to detect E. histolytica in spiked tap water samples, while DNA extraction after concentrating amoeba cells was required for spiked reservoir raw water samples. The detection limit of 50 E. histolytica cells in 100 ml tap water was achieved in 2 h from sample collection to real-time PCR data readout. With these established protocols, 78 tap water samples, 11 reservoir raw water samples and 4 feed water samples from Singapore water supply systems were analyzed by both conventional PCR and real-time PCR methods. No E. histolytica cell was detected in tested samples.

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Multidrug-Resistant Bacteria Isolated from Different Aquatic Environments in the North of Spain and South of France

Microorganisms ◽

10.3390/microorganisms8091425 ◽

2020 ◽

Vol 8 (9) ◽

pp. 1425

Author(s):

Lara Pérez-Etayo ◽

David González ◽

José Leiva ◽

Ana Isabel Vitas

Keyword(s):

Wastewater Treatment Plants ◽

Multidrug Resistant ◽

World Health ◽

Resistant Bacteria ◽

Aquatic Environments ◽

Multidrug Resistant Bacteria ◽

Antibiotic Resistant ◽

The North ◽

New Antibiotics ◽

Health Organization

Due to the global progress of antimicrobial resistance, the World Health Organization (WHO) published the list of the antibiotic-resistant “priority pathogens” in order to promote research and development of new antibiotics to the families of bacteria that cause severe and often deadly infections. In the framework of the One Health approach, the surveillance of these pathogens in different environments should be implemented in order to analyze their spread and the potential risk of transmission of antibiotic resistances by food and water. Therefore, the objective of this work was to determine the presence of high and critical priority pathogens included in the aforementioned list in different aquatic environments in the POCTEFA area (North Spain–South France). In addition to these pathogens, detection of colistin-resistant Enterobacteriaceae was included due its relevance as being the antibiotic of choice to treat infections caused by multidrug resistant bacteria (MDR). From the total of 80 analyzed samples, 100% of the wastewater treatment plants (WWTPs) and collectors (from hospitals and slaughterhouses) and 96.4% of the rivers, carried antibiotic resistant bacteria (ARB) against the tested antibiotics. Fifty-five (17.7%) of the isolates were identified as target microorganisms (high and critical priority pathogens of WHO list) and 58.2% (n = 32) of them came from WWTPs and collectors. Phenotypic and genotypic characterization showed that 96.4% were MDR and resistance to penicillins/cephalosporins was the most widespread. The presence of bla genes, KPC-type carbapenemases, mcr-1 and vanB genes has been confirmed. In summary, the presence of clinically relevant MDR bacteria in the studied aquatic environments demonstrates the need to improve surveillance and treatments of wastewaters from slaughterhouses, hospitals and WWTPs, in order to minimize the dispersion of resistance through the effluents of these areas.

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Detection of Virulence Genes in Multidrug Resistant Enterococci Isolated from Feedlots Dairy and Beef Cattle: Implications for Human Health and Food Safety

BioMed Research International ◽

10.1155/2019/5921840 ◽

2019 ◽

Vol 2019 ◽

pp. 1-13 ◽

Cited By ~ 5

Author(s):

Frank Eric Tatsing Foka ◽

Collins Njie Ateba

Keyword(s):

Public Health ◽

Antibiotic Resistance ◽

Virulence Genes ◽

Efflux Pump ◽

Health Sector ◽

Multidrug Resistant ◽

North West ◽

The North ◽

Vancomycin Resistant ◽

The Impact

The misuse/abuse of antibiotics in intensive animal rearing and communities led to the emergence of resistant isolates such as vancomycin-resistant enterococci (VREs) worldwide. This has become a major source of concern for the public health sector. The aim of this study was to report the antibiotic resistance profiles and to highlight the presence of virulence genes in VREs isolated from feedlots cattle of the North-West Province of South Africa. 384 faecal samples, 24 drinking troughs water, and 24 soil samples were collected aseptically from 6 registered feedlots. Biochemical and molecular methods were used to identify and categorise the enterococci isolates. Their antibiotic resistance profiles were assessed and genotypic methods were used to determine their antibiotic resistance and their virulence profiles. 527 presumptive isolates were recovered, out of which 289 isolates were confirmed asEnterococcussp. Specifically,E. faecalis(9%),E. faecium(10%),E. durans(69%),E. gallinarum(6%),E. casseliflavus(2%),E. mundtii(2%), andE. avium(2%) were screened after molecular assays.VanA(62%),vanB(17%), andvanC(21%) resistance genes were detected in 176Enterococcussp., respectively. Moreover,tetK(26),tetL(57),msrA/B(111), andmefA(9) efflux pump genes were detected in 138 VRE isolates.Multiple antibiotic resistances were confirmed in all the VRE isolates of this study; the most common antibiotic resistance phenotype wasTETR-AMPR-AMXR-VANR-PENR-LINR-ERYR.CylA,hyl,esp,gelE, andasa1virulence genes were detected in 86 VREs with the exception of vancomycin-resistantE. mundtiiisolates that did not display any virulence factor. Most VRE isolates had more than one virulence genes but the most encountered virulence profile wasgelE-hyl. Potentially pathogenic multidrug resistant VREs were detected in this study; this highlights the impact of extensive usage of antimicrobials in intensive animal rearing and its implications on public health cannot be undermined.

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Molecular characteristics of the multidrug-resistant mycobacterium tuberculosis strains in the North-west Russia

Molecular Genetics Microbiology and Virology (Russian version) ◽

10.18821/0208-0613-2016-34-1-30-33 ◽

2016 ◽

Vol 34 (1) ◽

pp. 30 ◽

Cited By ~ 1

Author(s):

A. A. Vyazovaya ◽

I. V. Mokrousov ◽

V. Yu. Zhuravlev ◽

N. S. Solovieva ◽

T. F. Otten ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Multidrug Resistant ◽

Molecular Characteristics ◽

North West ◽

The North

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Molecular evidence for the ubiquitous presence of Legionella species in Dutch tap water installations

Journal of Water and Health ◽

10.2166/wh.2007.033 ◽

2007 ◽

Vol 5 (3) ◽

pp. 375-383 ◽

Cited By ~ 23

Author(s):

Bram M. W. Diederen ◽

Caroline M. A. de Jong ◽

Ingrid Aarts ◽

Marcel F. Peeters ◽

Anneke van der Zee

Keyword(s):

16S Rrna ◽

Water Samples ◽

Tap Water ◽

Molecular Evidence ◽

Rrna Gene ◽

Specific Probe ◽

Detection Rates ◽

Specific Pcr ◽

Legionella Species ◽

Species Specific

Our aim was to investigate the occurrence and identity of Legionella spp. in Dutch tap water installations using culture, real-time PCR and sequence analysis. The PCR assays used were a 16S rRNA gene based PCR with both a Legionella species specific probe and a L. pneumophila specific probe and a L. pneumophila-specific PCR based on the sequence of the mip gene. A total of 357 water samples from 250 locations in The Netherlands was investigated. The detection rates of Legionella spp. were 2,2% (8 of 357) by culture, and 87,1% (311 of 357) by PCR. The majority of samples was found to contain Legionella species other than L. pneumophila. These comprised of Legionella Like Amoebal Pathogens (LLAPs), L. busanensis, L. worsliensis and others. Fourteen (3,9%) samples were positive for L. pneumophila by either culture, 16S rRNA based PCR and/or mip based PCR. It is apparent from this study that Legionella spp. DNA is ubiquitous in Dutch potable water samples. Our findings further suggest that LLAPs and viable but nonculturable (VBNC) Legionella represent a large proportion of the population in man-made environments.

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Genomic analysis of prevalent Staphylococcus epidermidis multidrug-resistant strains isolated during eight years in a single children hospital in México City.

10.7287/peerj.preprints.27693 ◽

2019 ◽

Author(s):

Roberto Cabrera-Contreras ◽

Rosa I Santamaría ◽

Patricia Bustos ◽

Irma Martínez-Flores ◽

Enrique Meléndez ◽

...

Keyword(s):

Mexico City ◽

Staphylococcus Epidermidis ◽

Biofilm Formation ◽

Genomic Analysis ◽

Multidrug Resistant ◽

Whole Genome ◽

Antibiotic Resistant ◽

Transfer Rates ◽

Children Health ◽

Health Care Unit

Staphylococcus epidermidis is a human commensal and pathogen worldwide distributed. In this work, we surveyed for multi-resistant S. epidermidis strains in eight years at a children health-care unit in México City. Multidrug-resistant S. epidermidis were present in all years of the study. Resistance to methicillin, beta-lactams, fluoroquinolones, and macrolides were included. To understand the genetic basis of antibiotic resistance and its association with virulence and gene exchange, we sequenced the genomes of 17 S. epidermidis isolates. Whole-genome nucleotide identities between all the pairs of S. epidermidis strains were about 97% to 99%. We inferred a clonal structure and eight Multilocus Sequence Types (MLST´s) in the S. epidermidis sequenced collection. The profile of virulence includes genes involved in biofilm formation and phenol-soluble modulins (PSMs). However, half of the S. epidermidis analyzed lacked the icaoperon for biofilm formation. Likely, they are commensal S. epidermidis strains but multi-antibiotic resistant. Uneven distribution of insertion sequences, phages, and CRISPR-Cas immunity phage systems suggest frequent horizontal gene transfer. Rates of recombination between S. epidermidis strains were more prevalent than the mutation rate and affected the whole genome. Therefore, the multidrug-resistance, independently of the pathogenic traits, might explain the persistence of specific highly adapted S. epidermidis clonal lineages in nosocomial settings.

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EVALUATION OF EFFLUX PUMP ACTIVITY AND BIOFILM FORMATION IN MULTIDRUG RESISTANT KLEBSIELLA PNEUMONIAE ISOLATES IN TANTA, EGYPT

International Research Journal of Pharmacy ◽

10.7897/2230-8407.1203123 ◽

2021 ◽

Vol 12 (3) ◽

pp. 1-5

Author(s):

Tarek El-Said El-Banna ◽

Fatma Ibrahim Sonbol ◽

Heba M El-Dawy ◽

Lamiaa A Al-Madboly

Keyword(s):

Drug Resistance ◽

Klebsiella Pneumoniae ◽

Biofilm Formation ◽

Efflux Pump ◽

Treatment Options ◽

Multidrug Resistant ◽

High Morbidity ◽

Biochemical Tests ◽

Antibiotic Resistance Profile ◽

Pump Activity

Nosocomial and community acquired infections that caused by multidrug-resistant (MDR) Klebsiella pneumoniae isolates are widespread recently resulting in high morbidity and mortality due to limited number of treatment options with effective antibiotics. The aim of this study is to evaluate the antibiotic resistance profile, biofilm formation and efflux pump activity of MDR K. pneumoniae isolates collected from different hospitals in Tanta, Egypt. A total of 70 K. pneumoniae isolates characterized by standard biochemical tests and confirmed by MALDI-TOF/MS were screened for antibiotic susceptibility, efflux pump activity and biofilm formation. Isolates displayed high resistance to penicillins, cephalosporins, trimethoprim-sulfamethoxazole and the majority of tested fluoro/-quinolones and decreased resistance to imipenem, amikacin, chloramphenicol, tigecycline and colistin. Out of 70 K. pneumoniae isolates, 2 isolates exhibited Pan Drug-Resistance (PDR) profile while 57 (81.4%) and 11 (15.7%) exhibited MDR and Extensively drug-resistance (XDR) profiles, respectively. Sixty-four (91.4%) isolates exhibited efflux pump activity while all tested isolates had the ability to form biofilm with varied degrees as 40 (57.1%), 26 (37.1%), and 4 (5.7%) isolates were strong, moderate and weak biofilm producers, respectively. Also, a strong relation between efflux pump activity and biofilm formation per isolate was detected. In conclusion, Multidrug resistance, biofilm formation and efflux pump capabilities in K. pneumoniae have serious public health implications in the management and control of infections caused by this bacterium. Therefore, a multifaceted approach and precise planning are recommended in controlling these infections

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Biochemical and Molecular Analysis ofStaphylococcus aureusClinical Isolates from Hospitalized Patients

Canadian Journal of Infectious Diseases and Medical Microbiology ◽

10.1155/2016/9041636 ◽

2016 ◽

Vol 2016 ◽

pp. 1-7 ◽

Cited By ~ 9

Author(s):

Amit Karmakar ◽

Parimal Dua ◽

Chandradipa Ghosh

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Agents ◽

Multidrug Resistant ◽

Hospitalized Patients ◽

Clinical Samples ◽

Resistance Pattern ◽

Biochemical Tests ◽

Species Specific ◽

High Level ◽

Gelatin Hydrolysis

Staphylococcus aureusis opportunistic human as well as animal pathogen that causes a variety of diseases. A total of 100Staphylococcus aureusisolates were obtained from clinical samples derived from hospitalized patients. The presumptiveStaphylococcus aureusclinical isolates were identified phenotypically by different biochemical tests. Molecular identification was done by PCR using species specific 16S rRNA primer pairs and finally 100 isolates were found to be positive asStaphylococcus aureus. Screened isolates were further analyzed by several microbiological diagnostics tests including gelatin hydrolysis, protease, and lipase tests. It was found that 78%, 81%, and 51% isolates were positive for gelatin hydrolysis, protease, and lipase activities, respectively. Antibiogram analysis of isolatedStaphylococcus aureusstrains with respect to different antimicrobial agents revealed resistance pattern ranging from 57 to 96%. Our study also shows 70% strains to be MRSA, 54.3% as VRSA, and 54.3% as both MRSA and VRSA. All the identified isolates were subjected to detection ofmecA,nuc, andhlbgenes and 70%, 84%, and 40% were found to harbourmecA,nuc, andhlbgenes, respectively. The current investigation is highly important and informative for the high level multidrug resistantStaphylococcus aureusinfections inclusive also of methicillin and vancomycin.

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