Characterization of biofilms and antimicrobial resistance of coagulase-negative Staphylococcus species involved with subclinical mastitis

2021 ◽  
pp. 1-6
Author(s):  
Márcia Silva Francisco ◽  
Ciro César Rossi ◽  
Maria Aparecida Vasconcelos Paiva Brito ◽  
Marinella Silva Laport ◽  
Elaine Menezes Barros ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Biofilm Formation ◽  
Subclinical Mastitis ◽  
Penicillin G ◽  
Coagulase Negative Staphylococcus ◽  
Central Feature ◽  
High Genetic Diversity ◽  
Dairy Animals

Abstract Biofilm formation is a central feature to guarantee staphylococcal persistence in hosts and is associated with several diseases that are difficult to treat. In this research paper, biofilm formation and antimicrobial susceptibility were investigated in staphylococcal strains belonging to several species. These strains were isolated from the milk of cows with subclinical mastitis and most of them were coagulase-negative, with the prevalence of Staphylococcus chromogenes. High genetic diversity was observed among the strains by pulsed field gel electrophoresis. Antimicrobial resistance was assessed by disk diffusion and more than 50% of the strains were resistant to ampicillin and penicillin G, with multi-resistance profiles (13.6%) also being observed. Most strains (65.9%) formed biofilms when cultivated in BHI supplemented with 1% glucose. Most strains (72.7%) carried the intercellular adhesion gene (icaA), while less than half (36.3%) carried the biofilm-associated protein gene (bap). Concentrations of up to 10xMIC of erythromycin and tetracycline were not sufficient to suppress cell viability in preformed biofilms. Our results revealed that a genetically diverse group of biofilm-forming Staphylococcus species can be involved in subclinical mastitis. Since high antimicrobial concentrations cannot eradicate biofilm cells in vitro, their use in dairy animals may be ineffective in controlling infections, while supporting selection of resistant microorganisms. These data reinforce the need for alternative therapies aiming at disrupting biofilms for effective disease control.

scholarly journals Antimicrobial resistance, virulence genes and biofilm formation in Enterococcus species isolated from milk of sheep and goat with subclinical mastitis

PLoS ONE ◽  
2021 ◽  
Vol 16 (11) ◽  
pp. e0259584
Author(s):  
Mona A. El-Zamkan ◽  
Hams M. A. Mohamed
Keyword(s):  
Multidrug Resistance ◽  
Antimicrobial Resistance ◽  
Biofilm Formation ◽  
Virulence Genes ◽  
Subclinical Mastitis ◽  
Virulence Determinants ◽  
Antimicrobial Susceptibility Test ◽  
Rdna Sequencing ◽  
Dairy Animals ◽  
Mastitic Milk

This study is designed to discuss the antimicrobial resistance, virulence determinants and biofilm formation capacity of Enterococcus spp. isolated from milk of sheep and goat with subclinical mastitis in Qena, Egypt. The obtained isolates were identified by the VITEK2 system and 16S rDNA sequencing as E. faecalis, E. faecium, E. casseliflavus and E. hirae. Overall, E. faecalis and E. faecium were the dominant species recovered from mastitic milk samples. The antimicrobial susceptibility test evidenced multidrug resistance of the isolates against the following antimicrobials: oxacillin (89.2.%), followed by vancomycin (75.7%) and linezolid (70.3%). Also, most of these isolates (73%) could form biofilms. For example, 18.9% of Enterococcus strains formed strong biofilm, whereas 32.4% of isolates formed moderate biofilm and 21.6% of isolates formed weak biofilm. The most prevalent resistance genes found in our isolates were blaZ (54%), vanA (40%), ermB (51.4%), tetM (13.5%) and optrA (10.8%). Moreover, asa1 (37.8%), cylA (42.3%), gelE (78.4%), esp (32.4%), EF3314(48.6%) and ace (75.5%) were the most common virulence genes. A significant correlation was found between biofilm formation, multidrug resistance and virulence genes of the isolates. This study highlights several aspects of virulence and harmfulness of Enterococcus strains isolated from subclinical mastitic milk, which necessitates continuous inspection and monitoring of dairy animals.

scholarly journals Antimicrobial resistance patterns and biofilm formation of coagulase-negative Staphylococcus species isolated from subclinical mastitis cow milk samples submitted to the Onderstepoort Milk Laboratory

2019 ◽  
Vol 15 (1) ◽  
Author(s):  
Lufuno Phophi ◽  
Inge-Marie Petzer ◽  
Daniel Nenene Qekwana
Keyword(s):  
Antimicrobial Resistance ◽  
Biofilm Formation ◽  
Subclinical Mastitis ◽  
Multidrug Resistant ◽  
Clinical Mastitis ◽  
Cow Milk ◽  
Coagulase Negative Staphylococcus ◽  
Milk Samples ◽  
Significant Difference ◽  
Resistance Patterns

Abstract Background Increased prevalence of antimicrobial resistance, treatment failure, and financial losses have been reported in dairy cows with coagulase-negative Staphylococcus (CoNS) clinical mastitis, however, studies on CoNS infections are limited in South Africa. Therefore, the objectives of this study were to investigate the antimicrobial resistance patterns and biofilm formation in CoNS isolated from cow milk samples submitted to the Onderstepoort Milk Laboratory. Results A total of 142 confirmed CoNS isolates were used for this study. Biofilm formation was identified in 18% of CoNS tested. Staphylococcus chromogenes (11%) had the highest proportion of biofilm formation followed by S. haemolyticus (4.0%), S. epidermidis, S. hominis, S. xylosus, and S. simulans with 1% respectively. Ninety percent (90%) of CoNS were resistant to at least one antimicrobial (AMR) and 51% were multidrug-resistant (MDR). Resistance among CoNS was the highest to ampicillin (90%) and penicillin (89%), few isolates resistant to cefoxitin and vancomycin, 9% respectively. Similarly, MDR-S. haemolyticus (44%), MDR-S. epidermidis (65%), and MDR-S. chromogenes (52%) were mainly resistant to penicillins. The most common resistance patterns observed were resistance to penicillin-ampicillin (16%) and penicillin-ampicillin-erythromycin (10%). Only 42% of biofilm positive CoNS were MDR. Conclusion The majority of CoNS in this study were resistance to penicillins. In addition, most isolates were β-lactam resistant and MDR. Biofilm formation among the CoNS in this study was uncommon and there was no significant difference in the proportion of MDR-CoNS based on the ability to form a biofilm.

scholarly journals Expression and Characterization of the Flocculin Flo11/Muc1, a Saccharomyces cerevisiae Mannoprotein with Homotypic Properties of Adhesion

2007 ◽  
Vol 6 (12) ◽  
pp. 2214-2221 ◽  
Author(s):  
Lois M. Douglas ◽  
Li Li ◽  
Yang Yang ◽  
A. M. Dranginis
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Biofilm Formation ◽  
In Vitro System ◽  
Glycosylphosphatidylinositol Anchor ◽  
Fungal Adhesion ◽  
Very High ◽  
Molecular Weight Form

ABSTRACT The Flo11/Muc1 flocculin has diverse phenotypic effects. Saccharomyces cerevisiae cells of strain background Σ1278b require Flo11p to form pseudohyphae, invade agar, adhere to plastic, and develop biofilms, but they do not flocculate. We show that S. cerevisiae var. diastaticus strains, on the other hand, exhibit Flo11-dependent flocculation and biofilm formation but do not invade agar or form pseudohyphae. In order to study the nature of the Flo11p proteins produced by these two types of strains, we examined secreted Flo11p, encoded by a plasmid-borne gene, in which the glycosylphosphatidylinositol anchor sequences had been replaced by a histidine tag. A protein of approximately 196 kDa was secreted from both strains, which upon purification and concentration, aggregated into a form with a very high molecular mass. When secreted Flo11p was covalently attached to microscopic beads, it conferred the ability to specifically bind to S. cerevisiae var. diastaticus cells, which flocculate, but not to Σ1278b cells, which do not flocculate. This was true for the 196-kDa form as well as the high-molecular-weight form of Flo11p, regardless of the strain source. The coated beads bound to S. cerevisiae var. diastaticus cells expressing FLO11 and failed to bind to cells with a deletion of FLO11, demonstrating a homotypic adhesive mechanism. Flo11p was shown to be a mannoprotein. Bead-to-cell adhesion was inhibited by mannose, which also inhibits Flo11-dependent flocculation in vivo, further suggesting that this in vitro system is a useful model for the study of fungal adhesion.

scholarly journals Enterotoxins production, biofilm formation and antimicrobial resistance of Staphylococcus aureus strains isolated from refrigerated raw cow milk

2019 ◽  
Vol 42 ◽  
pp. e45231
Author(s):  
Camila Lampugnani ◽  
Maike Taís Maziero Montanhini ◽  
Maria Emilene Martino Campos‐Galvão ◽  
Luis Augusto Nero ◽  
Luciano dos Santos Bersot
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Resistance ◽  
Biofilm Formation ◽  
Raw Milk ◽  
Cow Milk ◽  
In Vitro Assays ◽  
Vitro Assay ◽  
Milk Samples ◽  
Adherence Ability

This study aimed to isolate Staphylococcus aureus in refrigerated raw cow milk, and identify the presence of enterotoxin-expression genes, enterotoxin production and adherence ability, and antimicrobial resistance potential of the isolated strains. Fifty raw milk samples obtained in different dairy farms were analyzed for S. aureus and evaluated in the isolates the presence of genes associated with the production of major staphylococcal enterotoxins and biofilm formation. In vitro assays were also performed to evaluate the production of enterotoxins and adherence ability, and the antimicrobial resistance. One half (25/50) of raw milk samples presented coagulase-positive staphylococci and 95.2% of the isolates were confirmed to be S. aureus. Among them, 42.4% were carrying genes for enterotoxins production; however, only one isolate was able to produce enterotoxins. All S. aureus isolates were carrying at least two genes associated with biofilm formation and 95.2% isolates was able to adhere upon the in vitro assay. All isolates demonstrated antimicrobial resistance potential to one or more of the tested antibiotics.

Effects of vancomycin, daptomycin, and tigecycline on coagulase-negative staphylococcus biofilm and bacterial viability within biofilm: an in vitro biofilm model

2016 ◽  
Vol 62 (9) ◽  
pp. 735-743 ◽  
Author(s):  
Barcin Ozturk ◽  
Necati Gunay ◽  
Bulent M. Ertugrul ◽  
Serhan Sakarya
Keyword(s):  
Biofilm Formation ◽  
Treatment Time ◽  
Antimicrobial Effect ◽  
Bacterial Biofilm ◽  
Coagulase Negative Staphylococcus ◽  
Biofilm Inhibition ◽  
Major Barrier ◽  
Biofilm Model ◽  
Repeated Doses

Bacteria may hide in a hydrated polysaccharide matrix known as a biofilm. The structure of the bacterial biofilm renders phagocytosis difficult and increases antibiotic resistance. We hypothesized that repeated doses of antibiotics have an effect on bacteria within the biofilm and that it could inhibit or eradicate biofilm formation. Two clinical biofilm-positive coagulase-negative staphylococcus isolates were evaluated. The effects of antibiotics on preformed and nascent biofilm and on bacterial eradication within the biofilm were determined using different doses of vancomycin, daptomycin, and tigecycline for different durations in an in vitro biofilm model. Vancomycin neither penetrated the biofilm nor had any microbicidal effect on bacteria within the biofilm. Daptomycin had a microbicidal effect on bacteria within the biofilm but had no effect on biofilm inhibition and eradication (independent from dose and treatment time). Tigecycline inhibited and eradicated biofilm formation and had a microbicidal effect on bacteria within the biofilm. In conclusion, (i) biofilm formation appeared to be a major barrier to vancomycin activity, (ii) daptomycin had an antimicrobial effect on the bacteria within the biofilm but not on the biofilm burden, and (iii) tigecycline had effects both on bacteria within the biofilm and on biofilm burden. Thus, both tigecycline and daptomycin might be promising candidates for the treatment of biofilm infections.

scholarly journals In Vitro Characterization of a Biaryl Amide Anti-virulence Compound Targeting Candida albicans Filamentation and Biofilm Formation

2018 ◽  
Vol 8 ◽  
Author(s):  
Jesus A. Romo ◽  
Christopher G. Pierce ◽  
Marisol Esqueda ◽  
Chiung-Yu Hung ◽  
Stephen. P. Saville ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Biofilm Formation ◽  
In Vitro Characterization

scholarly journals Antimicrobial Resistance, Biofilm Formation and mecA Characterization of Methicillin-Susceptible S. aureus and Non-S. aureus of Beef Meat Origin in Egypt

2016 ◽  
Vol 7 ◽  
Author(s):  
Kamelia M. Osman ◽  
Aziza M. Amer ◽  
Jihan M. Badr ◽  
Nashwa M. Helmy ◽  
Rehab A. Elhelw ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Biofilm Formation ◽  
Beef Meat

scholarly journals Isolation, Biochemical and Molecular Identification, and In-Vitro Antimicrobial Resistance Patterns of Bacteria Isolated from Bubaline Subclinical Mastitis in South India

PLoS ONE ◽  
2015 ◽  
Vol 10 (11) ◽  
pp. e0142717 ◽  
Author(s):  
P. L. Preethirani ◽  
Shrikrishna Isloor ◽  
S. Sundareshan ◽  
V. Nuthanalakshmi ◽  
K. Deepthikiran ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Molecular Identification ◽  
South India ◽  
Subclinical Mastitis ◽  
Resistance Patterns

scholarly journals Bovine-associated non-aureus staphylococci suppress Staphylococcus aureus biofilm dispersal in vitro yet not through agr regulation

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Bruno Toledo-Silva ◽  
Fernando N. de Souza ◽  
Kristien Mertens ◽  
Sofie Piepers ◽  
Freddy Haesebrouck ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Communication Systems ◽  
Bovine Milk ◽  
Subclinical Mastitis ◽  
Interspecies Interactions ◽  
Environmental Signals ◽  
Biofilm Dispersion ◽  
Biofilm Development

AbstractBiofilm formation is a significant virulence factor in Staphylococcus (S.) aureus strains causing subclinical mastitis in dairy cows. A role of environmental signals and communication systems in biofilm development, such as the agr system in S. aureus, is suggested. In the context of multispecies biofilm communities, the presence of non-aureus staphylococci (NAS) might influence S. aureus colonization of the bovine mammary gland, yet, such interspecies interactions have been poorly studied. We determined whether 34 S. chromogenes, 11 S. epidermidis, and 14 S. simulans isolates originating from bovine milk samples and teat apices (TA) were able to affect biofilm formation and dispersion of S. aureus, and if so, how isolate traits such as the capacity to regulate the S. aureus agr quorum sensing system are determinants in this process. The capacity of an agr-positive S. aureus strain to form biofilm was increased more in the presence of S. chromogenes than in the presence of S. simulans and S. epidermidis isolates and in the presence of NAS isolates that do not harbor biofilm related genes. On the other hand, biofilm dispersion of this particular S. aureus strain was suppressed by NAS as a group, an effect that was more pronounced by isolates from TA. Furthermore, the observed effects on biofilm formation and dispersion of the agr-positive S. aureus strain as well as of an agr-negative S. aureus strain did not depend on the capacity of NAS to repress the agr system.

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