Virulence Genes, Biofilm Formation, and Antimicrobial Resistance of Enterococcus Faecalis and Enterococcus Faecium Isolated from Bovine Mastitis Milk Samples from 2015 to 2020

2021 ◽  
Author(s):  
Hyeon-Jin Kim ◽  
Hye-Young Youn ◽  
Hye-Jeong Kang ◽  
Jin-San Moon ◽  
Kun-Ho Seo
Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 661
Author(s):  
Sunghyun Yoon ◽  
Young Ju Lee

Enterococci are considered to be environmental mastitis-causing pathogens that can easily spread antimicrobial resistance or virulence genes via horizontal transfer. In this study, the molecular characteristics of enterococci from bulk tank milk were investigated to assess the importance of dairy herd management. A total of 338 enterococci (305 Enterococcus faecalis and 33 Enterococcus faecium) were isolated from 1584 batches of bulk tank milk samples from 396 farms affiliated with four dairy companies in Korea, and significant differences (40.6–79.7%) (p < 0.05) in the prevalence of enterococci were observed in the samples from different companies. Enterococci showed the highest resistance to tetracycline (TET) (73.4%), followed by doxycycline (DOX) (49.7%) and erythromycin (ERY) (46.2%), while two enterococci isolates showed resistance to vancomycin (VAN). Among 146 tetracycline (TET) and ERY-resistant enterococci, each 50 (19.4%) enterococci carried combination-resistance and transposon gene types erm(B) + tet(M) + IntTn and erm(B) + tet(L) + tet(M) + IntTn, respectively. The virulence genes such as ace (99.0%), efaA (97.7%), cad1 (95.7%), and gelE (85.9%) were highly conserved in E. faecalis and significantly predominated over E. faecium (p < 0.001). Our results indicate that pathogens from bulk tank milk can also become a reservoir for the dissemination of antimicrobial resistance and virulence factors through cross-contamination processes.


2009 ◽  
Vol 27 (Special Issue 1) ◽  
pp. S354-S356 ◽  
Author(s):  
L. Necidová ◽  
B. Janštová ◽  
S. Karpíšková ◽  
Š. Cupáková ◽  
M. Dušková ◽  
...  

The aim of this study was to monitore the capability of <I>Enterococcus fecalis</I> and <I>Enterococcus fecium</I> to form biofilms. Enterococci isolates originated from individual milk, bulk milk samples and environmental swabs obtained at farm level, dairy plant level including semi and final dairy products. Biofilm formation potential was determined by growing the tested strains in glas tubes containing BHI medium. The capability of forming biofilms was detected in 28% of <I>Enterococcus</I> spp. strains. Higher number of biofilm forming strains of the <I>Enterococcus faecium</I> (33%) than <I>Enterococcus faecalis</I> (28%) has been registered. Isolates obtained at plant level were forming biofilms more often than isolates from plant level and in final products (cheese and curd cheese), no isolate has been seen to be able to form biofilm.


2007 ◽  
Vol 53 (3) ◽  
pp. 372-379 ◽  
Author(s):  
N. Klibi ◽  
K. Ben Slama ◽  
Y. Sáenz ◽  
A. Masmoudi ◽  
S. Zanetti ◽  
...  

Phenotypic and genotypic determination of virulence factors were carried out in 46 high-level gentamicin-resistant (HLGR) clinical Enterococcus faecalis (n = 34) and Enterococcus faecium (n = 12) isolates recovered from different patients in La Rabta Hospital in Tunis, Tunisia, between 2000 and 2003 (all these isolates harboured the aac(6′)–aph(2″) gene). The genes encoding virulence factors (agg, gelE, ace, cylLLS, esp, cpd, and fsrB) were analysed by PCR and sequencing. The production of gelatinase and hemolysin, the adherence to caco-2 and hep-2 cells, and the capacity for biofilm formation were investigated in all 46 HLGR enterococci. The percentages of E. faecalis isolates harbouring virulence genes were as follows: gelE, cpd, and ace (100%); fsrB (62%); agg (56%); cylLLS (41.2%); and esp (26.5%). The only virulence gene detected among the 12 HLGR E. faecium isolates was esp (58%). Gelatinase activity was detected in 22 of the 34 E. faecalis isolates (65%, most of them with the gelE+–fsrB+ genotype); the remaining 12 isolates were gelatinase-negative (with the gelE+–fsrB– genotype and the deletion of a 23.9 kb fragment of the fsr locus). Overall, 64% of the cylLLS-containing E. faecalis isolates showed β-hemolysis. A high proportion of our HLGR E. faecalis isolates, in contrast to E. faecium, showed moderate or strong biofilm formation or adherence to caco-2 and hep-2 cells.


Author(s):  
Andrea Lauková ◽  
Anna Kandričáková ◽  
Eva Bino

This study investigated eight types of Slovak dry fermented meat products (salami and sausages) that are available on the market and were produced by three different producers in different regions of Slovakia. The total counts of enterococci in these products ranged from 2.0 up to 6.0 cfu/g (log10). Three species were identified among the 15 selected enterococcal strains; Enterococcus faecium (8 strains), Enterococcus faecalis (3) and Enterococcus hirae (4). They were hemolysis-negative (γ-hemolysis) with a biofilm-forming ability, which was evaluated as low-grade biofilm formation, susceptible to conventional antibiotics and mainly susceptible to lantibiotic bacteriocins, namely, gallidermin and nisin; they even showed a higher susceptibility to gallidermin than to nisin. They were also susceptible to enterocin–durancin, but most strains showed resistance to enterocin A/P. This study indicated that bacteriocins can play a key role in preventing and/or protecting from undesirable bacterial multiplication or contamination in the food industry and that they have great potential for further experimental applications.


Author(s):  
Jayaprabha Chockalingam ◽  
Akshaya Balasubramanian ◽  
Jai Santhosh K. Krishnasamy ◽  
Karthika Balasundaram ◽  
Kavin Kumar Sivasamy ◽  
...  

2020 ◽  
Vol 48 ◽  
Author(s):  
Érica Chaves Lucio ◽  
Gisele Veneroni Gouveia ◽  
Mateus Matiuzzi Da Costa ◽  
Mário Baltazar De Oliveira ◽  
Rinaldo Aparecido Mota ◽  
...  

Background: Bovine mastitis remains one of the health problems that cause the most damage to milk producers. The negative impact of mastitis is due to reduced milk production, early slaughter of females, reduced commercial value of the affected animals, losses in the genetic potential of the herd, expenses with medicines and veterinary medical assistance. Staphylococcus spp. stands out as the cause of this disease and has been able to remain in the mammary gland, becoming resistant to several antimicrobials. The aims of the present study were to characterize the phenotypes, genotypes and resistance profiles of Staphylococcus spp. isolates from bovine mastitis cases in the state of Pernambuco, Brazil.Materials, Methods & Results: These isolates were classified according to biochemical tests and the presence of the nuc gene.  The polymerase chain reaction (PCR) for amplification of the mecA and blaZ genes was used to analyze the genetic potentials of antimicrobial resistance. Isolates were also phenotypically tested for resistance to nine antimicrobials (ampicillin, doxicillin, erythromycin, gentamicin, rifampicin, cephalothin, amoxicillin, nalidixic acid and oxacillin). The genetic potentials for biofilm production were evaluated by the amplifications of the icaD, icaA and bap genes. The phenotypic test of gentian violet was used for biofilm formation analyzes. Ninety-three (93.0%) of the isolates among the Staphylococcus spp. samples were classified as Staphylococcus aureus. The lowest percentage of sensitivity observed was for amoxicillin (28.0%). All of the isolates were sensitive to erythromycin and gentamicin, and 15 (15%) exhibited sensitivity to all of the drugs tested. All of the isolates were negative for the mecA gene, and 36 (36%) were positive for blaZ. In the adhesion microplate tests, 44 (44%) of the isolates were capable of biofilm formation. Of these, seven (15.9%) were strong formers, whereas 16 (36.3%) and 21 (47.8%) were moderate and weak formers, respectively. The icaD gene was confirmed in 89 (89%) of the isolates. The icaA gene was confirmed in 61 (61%) samples, and the bap gene in 52 (52%) samples. One of the samples did not possess icaA, icaD or bap and exhibited moderate biofilm formation according to the microplate adherence test. Sixteen isolates simultaneously exhibited the three genes tested for biofilm production (icaA, icaD and bap) and were negative according to the microplate adherence test.Discussion:  The indiscriminate use of antibiotics to treat mastitis is a common practice in the study area, which may have contributed to the high proportion of herds (88.23%; 15/17) with multi-resistant isolates, constituting a selection factor for the dissemination of resistant bacteria among herds.  The absence of the mecA gene in the present study may be associated with the development of resistant bacteria through another mechanism, such as the overproduction of beta-lactamases. The results demonstrate that antimicrobial resistance occurs in Staphylococcus spp. that cause bovine mastitis in herds of Pernambuco and that these isolates have the a great capacity for biofilm formation. It is necessary to sensitize the professionals involved in the milk production chain of Brazil regarding the importance of the adequate use of antimicrobials for the treatment and control of mastitis, since studies in the country indicate the dissemination of resistant bacterial strains.


2019 ◽  
Vol 63 (1) ◽  
pp. 93-97 ◽  
Author(s):  
Hanna Różańska ◽  
Aleksandra Lewtak-Piłat ◽  
Maria Kubajka ◽  
Marcin Weiner

Abstract Introduction: The aim of the study was to evaluate the occurrence of enterococci in inflammatory secretions from mastitic bovine udders and to assess their antimicrobial resistance. Material and Methods: A total of 2,000 mastitic milk samples from cows were tested in 2014–2017. The isolation of enterococci was performed by precultivation in buffered peptone water, selective multiplication in a broth with sodium azide and cristal violet, and cultivation on Slanetz and Bartley agar. The identification of enterococci was carried out using Api rapid ID 32 strep kits. The antimicrobial susceptibility was evaluated using the MIC technique. Results: Enterococci were isolated from 426 samples (21.3%). Enterococcus faecalis was the predominant species (360 strains), followed by E. faecium (35 isolates), and small numbers of others. The highest level of resistance was observed to lincomycin, tetracycline, quinupristin/dalfopristin (Synercid), erythromycin, kanamycin, streptomycin, chloramphenicol, and tylosin. Single strains were resistant to vancomycin and ciprofloxacin. All isolates were sensitive to daptomycin. E. faecalis presented a higher level of resistance in comparison to E. faecium, except to nitrofurantoin. Conclusion: The results showed frequent occurrence of enterococci in mastitic cow’s milk and confirmed the high rate of their antimicrobial resistance.


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