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

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

International Journal of Microbiology ◽

10.1155/2020/8863370 ◽

2020 ◽

Vol 2020 ◽

pp. 1-15

Author(s):

Mpho Defney Maje ◽

Christ Donald Kaptchouang Tchatchouang ◽

Madira Coutlyne Manganyi ◽

Justine Fri ◽

Collins Njie Ateba

Keyword(s):

Water Samples ◽

Biofilm Formation ◽

Tap Water ◽

Multidrug Resistant ◽

Biochemical Tests ◽

Antibiotic Resistant ◽

North West ◽

The North ◽

C 50 ◽

Species Specific

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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Interaction of Vibrio to Biotic and Abiotic Surfaces: Relationship between Hydrophobicity, Cell Adherence, Biofilm Production, and Cytotoxic Activity

Surfaces ◽

10.3390/surfaces1010014 ◽

2018 ◽

Vol 1 (1) ◽

pp. 187-201 ◽

Cited By ~ 1

Author(s):

Faouzi Lamari ◽

Sadok Khouadja ◽

Sami Rtimi

Keyword(s):

Biofilm Formation ◽

Dicentrarchus Labrax ◽

Sea Bass ◽

Strong Relationship ◽

Bacterial Strains ◽

Abiotic Surfaces ◽

Multiplex Pcr Assay ◽

Capsule Production ◽

Species Specific ◽

Pathogenic Agents

Vibrio parahaemolyticus and Vibrio alginolyticus are important pathogenic agents for both humans and aquatic animals. Twenty-five bacterial strains were isolated from infected sea bass (Dicentrarchus labrax) on thiosulfate citrate bile salts sucrose (TCBS) agar plates. For the species-specific detection of V. alginolyticus and V. parahaemolyticus, a multiplex PCR assay using two collagenase-targeted primer pairs allows the detection of four strains of V. parahaemolyticus and three strains of V. alginolyticus. The seven identified isolates were partitioned for capsule production, hydrophobicity, adherence, biofilm formation, invasion, and cytotoxicity against Hep-2 cells. Two V. parahaemolyticus (Spa2 and Spa3) and one V. alginolyticus (Va01) were capsule producers developing almost black colonies on CRA, they showed a strong hydrophobicity using bacterial adhesion to hydrocarbons test (BATH), and were able to produce high biofilm. Isolates were able to adhere and invade Hep-2 cells and exhibited dissimilar levels of cytotoxicity in epithelial cells. This study shows the strong relationship between adhesion, biofilm formation, invasion and the cytotoxicity of Vibrio strains. Thus, we found a strong and significant positive correlation between different virulence properties of these isolates. The present study shows that bacterial contact with the cells as well as adhesion and invasion are essential steps to induce cytotoxicity. However, the invasion is seen to be a post adherence event.

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Phenotypical Comparison of Pseudomonas Aeruginosa Isolated from Human and Veterinary Samples; Impact of Host Adaptation on Infection Pathogenesis

10.21203/rs.3.rs-1170308/v1 ◽

2021 ◽

Author(s):

Stefan J. Kaiser ◽

Annalisa DeRosa ◽

Christa Ewers ◽

Frank Günther

Keyword(s):

Pseudomonas Aeruginosa ◽

Water Samples ◽

Biofilm Formation ◽

Polymorphonuclear Leukocytes ◽

Environmental Isolates ◽

Antibiotic Resistant ◽

Pet Owners ◽

Zoonotic Risk ◽

Genetically Determined

Abstract Purpose: Determinants of virulence in Pseudomonas aeruginosa vary strongly depending on its habitat. In this study, we analyzed these alterations depending on the host organism in isolates cultured from canine ears and compared it to clinical extended-spectrum antibiotic-resistant Pseudomonas aeruginosa isolates (XDR), clinical antibiotic-sensitive (non-XDR) from humans and environmental isolates (EI) analyzed during our first study in 2017. Methods: A total of 22 veterinary isolates cultured from canine ears (VET) were examined for spontaneous biofilm formation, stress response in biofilm formation induced by meropenem, in vitro fitness, susceptibility to human serum and polymorphonuclear leukocytes and the genetically determined virulence factors toxA, exoS, exoT, exoU, exoY, nan1, cif, lasA and lasB.Results: We observed significantly elevated spontaneous biofilm formation and serum susceptibility in VET isolates compared to EI and non-XDR isolates as well as significantly decreased in vitro fitness compared to XDR isolates. The VET isolates resembled most the XDR subgroup of isolates previously cultured from blood. Within the environmental isolates, we observed an increase of spontaneous biofilm formation and exoU presence in isolates cultured from community water samples over hospital water samples to pool samples.Conclusions: Considering the distinct differences in some features of the examined VET isolates, a higher degree of phenotypical adaption can be assumed. Increased biofilm formation seems to be a common and characteristic event in isolates adapted to a specific habitat. Therefore amplification of potentially more virulent Pseudomonas aeruginosa strains in domestic animals may lead to elevated zoonotic risk for example for pet owners.

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Exogenous Protein as an Environmental Stimuli of Biofilm Formation in Select Bacterial Strains

10.1101/683979 ◽

2019 ◽

Author(s):

Donna Ye ◽

Lekha Bapu ◽

Mariane Mota Cavalcante ◽

Jesse Kato ◽

Maggie Lauria Sneideman ◽

...

Keyword(s):

Biofilm Formation ◽

Pathogenic Bacteria ◽

Milk Protein ◽

Flavobacterium Columnare ◽

Bacterial Strains ◽

Fish Pathogens ◽

Cell Counts ◽

Exogenous Protein ◽

High Concentrations ◽

Species Specific

ABSTRACTA screening of environmental conditions that would elicit robust biofilm in a collection ofSerratia marcescensisolated from soil revealed that exogenous milk protein increased biofilm productivity up to ten-fold. A select screening of fish pathogens, freshwater and human isolates identified several other species that responded similarly to exogenous protein. The optimal protein concentration was species specific;S. marcescensat 5% milk protein,Aeromonassp. at 2-3%,Flavobacterium columnareat 1% andPseudomonas aeruginosaat 0.1-0.4%. Media supplemented with milk protein also increased the cell counts in biofilm as well as the protein incorporated into the biofilm matrix. These data suggest that relatively high concentrations of exogenous protein may serve as an environmental trigger for biofilm formation, particularly for pathogenic bacteria exposed to relatively high concentrations of protein in bodily fluids and mucosal surfaces.

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Thiol and Disulfide Containing Vancomycin Derivatives Against Bacterial Resistance and Biofilm Formation

10.1101/2021.08.23.457343 ◽

2021 ◽

Author(s):

Inga S. Shchelik ◽

Karl Gademann

Keyword(s):

Biofilm Formation ◽

Bacterial Infections ◽

Bacterial Resistance ◽

Resistant Bacteria ◽

Bacterial Strains ◽

Antibiotic Resistant ◽

Promising Strategy ◽

Vancomycin Resistant ◽

New Compounds ◽

Sulfur Containing

Antibiotic-resistant and biofilm-associated infections constitute a rapidly growing issue. The last resort antibiotic vancomycin is under threat, due to the increasing appearance of vancomycin resistant bacteria as well as the formation of biofilms. Herein, we report a series of novel vancomycin derivatives carrying thiol- and disulfide-containing moieties. The new compounds exhibited enhanced antibacterial activity against a broad range of bacterial strains, including vancomycin resistant microbes and Gram-negative bacteria. Moreover, all obtained derivatives demonstrated improved antibiofilm formation activity against VanB resistant Enterococcus compared to vancomycin. This work established a promising strategy for combating drug-resistant bacterial infections or disrupting biofilm formation and advances the knowledge on structural optimization of antibiotics with sulfur-containing modifications.

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Tryptophan Silver Nanoparticles Synthesized by Photoreduction Method: Characterization and Determination of Bactericidal and Anti-Biofilm Activities on Resistant and Susceptible Bacteria

International Journal of Tryptophan Research ◽

10.1177/1178646919831677 ◽

2019 ◽

Vol 12 ◽

pp. 117864691983167 ◽

Cited By ~ 7

Author(s):

Daniella dos Santos Courrol ◽

Carla Regina Borges Lopes ◽

Camila Bueno Pacheco Pereira ◽

Marcia Regina Franzolin ◽

Flávia Rodrigues de Oliveira Silva ◽

...

Keyword(s):

Silver Nanoparticles ◽

Biofilm Formation ◽

Antimicrobial Agents ◽

Irradiation Time ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Bacterial Strains ◽

Antibiotic Resistant ◽

E Coli ◽

Static Conditions

The high rates of antibiotics use in hospitals have resulted in a condition where multidrug-resistant pathogens have become a severe threat to the human health worldwide. Therefore, there is an increasing necessity to identify new antimicrobial agents that can inhibit the multidrug-resistant bacteria and biofilm formation. In this study, antibacterial and anti-biofilm activities of tryptophan silver nanoparticles (TrpAgNP) were investigated. The TrpAgNPs were synthesized by photoreduction method, and the influence of irradiation time and concentration of reagents were analyzed. The nanoparticles were characterized by transmission electron microscopy, Zeta Potential and (UV)-absorption spectra. The antibacterial activity of TrpAgNPs was tested for antibiotic-resistant and susceptible pathogens, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Citrobacter freundii, Klebsiella pneumoniae, Salmonella typhimurium, and Pseudomonas aeruginosa, evaluating the influence of photoreduction parameters in bactericidal effect. The results have shown that TrpAgNPs solutions with lower tryptophan/silver nitrate (AgNO3) ratio and higher AgNO3 concentration have higher bactericidal action against bacteria with inhibition of ~100% in almost all studied bacterial strains. The antimicrobial activity of TrpAgNPs within biofilms generated under static conditions of antibiotic-resistant and susceptible strains of S. aureus, S. epidermidis, E. coli, K. pneumoniae, C. freundii, and P. aeruginosa was also investigated. The results showed that TrpAgNPs have an inhibitory effect against biofilm formation, exceeding 50% in the case of Gram-negative bacteria ( E. coli, K. pneumoniae, C. freundii, and P. aeruginosa—54.8% to 98.8%). For Gram-positive species, an inhibition of biofilm formation of 68.7% to 72.2 % was observed for S. aureus and 20.0% to 40.2% for S. epidermidis.

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Prebiotic Combinations Effects on the Colonization of Staphylococcal Skin Strains

Microorganisms ◽

10.3390/microorganisms9010037 ◽

2020 ◽

Vol 9 (1) ◽

pp. 37

Author(s):

Silvia Di Lodovico ◽

Franco Gasparri ◽

Emanuela Di Campli ◽

Paola Di Fermo ◽

Simonetta D’Ercole ◽

...

Keyword(s):

Biofilm Formation ◽

Commensal Bacteria ◽

The Other ◽

Bacteriostatic Effect ◽

Skin Microbiota ◽

Relevant Effect ◽

Planktonic Phase ◽

Species Specific ◽

Biomass Quantification ◽

General Reduction

Background: An unbalanced skin microbiota due to an increase in pathogenic vs. commensal bacteria can be efficiently tackled by using prebiotics. The aim of this work was to identify novel prebiotic combinations by exerting species-specific action between S. aureus and S. epidermidis strains. Methods: First, the antimicrobial/antibiofilm effect of Xylitol-XYL and Galacto-OligoSaccharides–GOS combined with each other at different concentrations (1, 2.5, 5%) against S. aureus and S. epidermidis clinical strains was evaluated in time. Second, the most species-specific concentration was used to combine XYL with Fructo-OligoSaccharides–FOS, IsoMalto-Oligosaccharides–IMO, ArabinoGaLactan–LAG, inulin, dextran. Experiments were performed by OD600 detection, biomass quantification and LIVE/DEAD staining. Results: 1% XYL + 1% GOS showed the best species-specific action with an immediate antibacterial/antibiofilm action against S. aureus strains (up to 34.54% ± 5.35/64.68% ± 4.77) without a relevant effect on S. epidermidis. Among the other prebiotic formulations, 1% XYL plus 1% FOS (up to 49.17% ± 21.46/37.59% ± 6.34) or 1% IMO (up to 41.28% ± 4.88/36.70% ± 10.03) or 1% LAG (up to 38.21% ± 5.31/83.06% ± 5.11) showed antimicrobial/antibiofilm effects similar to 1% XYL+1% GOS. For all tested formulations, a prevalent bacteriostatic effect in the planktonic phase and a general reduction of S. aureus biofilm formation without loss of viability were recorded. Conclusion: The combinations of 1% XYL with 1% GOS or 1% FOS or 1% IMO or 1% LAG may help to control the balance of skin microbiota, representing good candidates for topic formulations.

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Bond Strengthening in Oral Bacterial Adhesion to Salivary Conditioning Films

Applied and Environmental Microbiology ◽

10.1128/aem.01119-08 ◽

2008 ◽

Vol 74 (17) ◽

pp. 5511-5515 ◽

Cited By ~ 40

Author(s):

Henny C. van der Mei ◽

Minie Rustema-Abbing ◽

Joop de Vries ◽

Henk J. Busscher

Keyword(s):

Bacterial Adhesion ◽

Biofilm Formation ◽

Specific Interactions ◽

Adhesion Forces ◽

Bacterial Strains ◽

Bacterial Interactions ◽

Initial Adhesion ◽

Conditioning Film ◽

Interacting Surfaces ◽

Conditioning Films

ABSTRACT Transition from reversible to irreversible bacterial adhesion is a highly relevant but poorly understood step in initial biofilm formation. We hypothesize that in oral biofilm formation, irreversible adhesion is caused by bond strengthening due to specific bacterial interactions with salivary conditioning films. Here, we compared the initial adhesion of six oral bacterial strains to salivary conditioning films with their adhesion to a bovine serum albumin (BSA) coating and related their adhesion to the strengthening of the binding forces measured with bacteria-coated atomic force microscopy cantilevers. All strains adhered in higher numbers to salivary conditioning films than to BSA coatings, and specific bacterial interactions with salivary conditioning films were accompanied by stronger initial adhesion forces. Bond strengthening occurred on a time scale of several tens of seconds and was slower for actinomyces than for streptococci. Nonspecific interactions between bacteria and BSA coatings strengthened twofold faster than their specific interactions with salivary conditioning films, likely because specific interactions require a closer approach of interacting surfaces with the removal of interfacial water and a more extensive rearrangement of surface structures. After bond strengthening, bacterial adhesion forces with a salivary conditioning film remained stronger than those with BSA coatings.

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Development of Effective Antibacterial Treatment: Lessons from the Past and Novel Approaches

Antibiotics ◽

10.3390/antibiotics10030230 ◽

2021 ◽

Vol 10 (3) ◽

pp. 230

Author(s):

Mariagrazia Di Luca ◽

Tiziano Marzo

Keyword(s):

Antibacterial Treatment ◽

Bacterial Strains ◽

Antibiotic Resistant ◽

The Past ◽

Novel Approaches ◽

Rapid Diffusion

In the last three decades, the appearance and rapid diffusion of antibiotic-resistant bacterial strains have been observed [...]

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1564. Activity of bacteriophage against multidrug resistant Klebsiella pneumoniae

Open Forum Infectious Diseases ◽

10.1093/ofid/ofaa439.1744 ◽

2020 ◽

Vol 7 (Supplement_1) ◽

pp. S782-S782

Author(s):

Sailaja Puttagunta ◽

Maya Kahan-Haanum ◽

Sharon Kredo-Russo ◽

Eyal Weinstock ◽

Efrat Khabra ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Unmet Need ◽

Multidrug Resistant ◽

Beta Lactamase ◽

Bacterial Strains ◽

Antibiotic Resistant ◽

Extended Spectrum Beta Lactamase ◽

Novel Approach ◽

Infection Types

Abstract Background The prevalence of extended-spectrum beta-lactamase (ESBL) producing and carbapenem resistant (CR) Klebsiella pneumoniae (KP) has significantly risen in all geographic regions. Infections due to these bacteria are associated with high mortality across different infection types. Even with newer options, there remains an unmet need for safe and effective therapeutic options to treat infections caused by ESBL and CR KP. Phage therapy offers a novel approach with an unprecedented and orthogonal mechanism of action for treatment of diseases caused by pathogenic bacterial strains that are insufficiently addressed by available antibiotics. Phage-based therapies confer a high strain-level specificity and have a strong intrinsic safety profile. Here we describe the identification of novel phages that can effectively target antibiotic resistant KP strains. Host range of the 21 phages on 33 strain KP panel via solid culture infectivity assays. Red marks resistance to infection while sensitivity to phage is marked in green Methods KP clinical strains were isolated from human stool specimens preserved in glycerol. Selective culturing was carried, followed by testing of individual colonies for motility, indole and urease production, sequenced and analyzed by Kleborate tool to determine antibiotic resistant genes. Natural phages were isolated from plaques that developed on susceptible bacterial targets, sequenced and characterized. Results Antibiotic-resistant KP strains encoding beta lactamase genes or a carbapenemase (n=33) were isolated from healthy individuals (n=3), and patients with inflammatory bowel disease (n=26) or primary sclerosing cholangitis (n=3). Isolates sequencing revealed bla CTX-M15 and/or bla SHV encoding strains and carbapenamase KPC-2. A panel of 21 phages targeting the beta-lactamase- and carbapenemase-producing KP strains were identified. Phage sequencing revealed that all phages belong to the Caudovirales order and include 6 Siphoviridae, 14 Myoviridae, and 1 Podoviridae. In vitro lytic activity of the phages was tested on the isolated bacteria and revealed a coverage of 70% of the 33 isolated antibiotic resistant strains, >50% of which were targeted by multiple phages. Conclusion Collectively, these results demonstrate the feasibility of identifying phage with potent activity against antibiotic resistant KP strains, and may provide a novel therapeutic approach for treatment of ESBL and CR KP infections. Disclosures All Authors: No reported disclosures

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