A standardized conjugation protocol to asses antibiotic resistance transfer between lactococcal species

Lampkowska, Joanna; Feld, Louise; Monaghan, Áine; Toomey, Niamh; Schjørring, Susanne; Jacobsen, B.; Voet, Hilko van der; Andersen, Sigrid Rita; Bolton, Declan; Aarts, H.J.M.; Krogfelt, Karen A.; Wilcks, Andrea; Bardowski, Jacek

doi:10.1016/j.ijfoodmicro.2008.06.017

Cited by 47 publications

(31 citation statements)

References 13 publications

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“…lactis and cremoris (Moreno et al, 1999;Flórez et al, 2008;Lampkowska et al, 2008). lactis and cremoris (Moreno et al, 1999;Flórez et al, 2008;Lampkowska et al, 2008).…”

Section: Transfer Of the Tet(s) Gene To L Monocytogenesunclassified

“…The ability to transfer genetic material via conjugation is widespread among Lactococcus species, and it has been demonstrated in L. lactis ssp. lactis and cremoris (Moreno et al, 1999;Flórez et al, 2008;Lampkowska et al, 2008). Several studies have described the transfer by conjugation of plasmids and transposons carrying tetracycline resistance genes from Enterococcus and Streptococcus to Listeria, and between Listeria species (Vicente et al, 1988;Facinelli et al, 1993;Perreten et al, 1997), but never from Lactococcus to Listeria.…”

Section: Transfer Of the Tet(s) Gene To L Monocytogenesmentioning

confidence: 99%

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Transfer of plasmid-mediated resistance to tetracycline in pathogenic bacteria from fish and aquaculture environments

Guglielmetti

Korhonen

Heikkinen

et al. 2009

FEMS Microbiology Letters

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The transferability of a large plasmid that harbors a tetracycline resistance gene tet(S), to fish and human pathogens was assessed using electrotransformation and conjugation. The plasmid, originally isolated from fish intestinal Lactococcus lactis ssp. lactis KYA-7, has potent antagonistic activity against the selected recipients (Lactococcus garvieae and Listeria monocytogenes), preventing conjugation. Therefore the tetracycline resistance determinant was transferred via electroporation to L. garvieae. A transformant clone was used as the donor in conjugation experiments with three different L. monocytogenes strains. To our knowledge, this is the first study showing the transfer of an antibiotic resistance plasmid from fish-associated lactic bacteria to L. monocytogenes, even if the donor L. garvieae was not the original host of the tetracycline resistance but experimentally created by electroporation. These results demonstrate that the antibiotic resistance genes in the fish intestinal bacteria have the potential to spread both to fish and human pathogens, posing a risk to aquaculture and consumer safety.

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“…lactis and cremoris (Moreno et al, 1999;Flórez et al, 2008;Lampkowska et al, 2008). lactis and cremoris (Moreno et al, 1999;Flórez et al, 2008;Lampkowska et al, 2008).…”

Section: Transfer Of the Tet(s) Gene To L Monocytogenesunclassified

Section: Transfer Of the Tet(s) Gene To L Monocytogenesmentioning

confidence: 99%

Transfer of plasmid-mediated resistance to tetracycline in pathogenic bacteria from fish and aquaculture environments

Guglielmetti

Korhonen

Heikkinen

et al. 2009

FEMS Microbiology Letters

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“…The ability of the lactococcal strains to transfer the tetracycline resistance genes to E. faecalis JH2-2, E. faecalis JH2-SS, L. lactis Bu2-60, and B. subtilis YBE01 was examined by the filter mating approach. The mating procedure was performed as described earlier (32). In short, exponentially growing donor and recipient strains were mixed in a total volume of 2 ml and poured onto a sterile membrane filter (HAWP04700; Millipore, Bedford, MA), which was incubated right side up for 18 to 20 h at 30°C, 37°C, or 42°C (depending on the recipient) on nonselective recipient-specific agar plates.…”

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confidence: 99%

Intra- and Interspecies Conjugal Transfer of Tn 916 -Like Elements from Lactococcus lactis In Vitro and In Vivo

Bogusławska

Zycka-Krzesinska

Wilcks

et al. 2009

Appl Environ Microbiol

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Tetracycline-resistant Lactococcus lactis strains originally isolated from Polish raw milk were analyzed for the ability to transfer their antibiotic resistance genes in vitro, using filter mating experiments, and in vivo, using germfree rats. Four of six analyzed L. lactis isolates were able to transfer tetracycline resistance determinants in vitro to L. lactis Bu2-60, at frequencies ranging from 10 ؊5 to 10 ؊7 transconjugants per recipient. Three of these four strains could also transfer resistance in vitro to Enterococcus faecalis JH2-2, whereas no transfer to Bacillus subtilis YBE01, Pseudomonas putida KT2442, Agrobacterium tumefaciens UBAPF2, or Escherichia coli JE2571 was observed. Rats were initially inoculated with the recipient E. faecalis strain JH2-2, and after a week, the L. lactis IBB477 and IBB487 donor strains were introduced. The first transconjugants were detected in fecal samples 3 days after introduction of the donors. A subtherapeutic concentration of tetracycline did not have any significant effect on the number of transconjugants, but transconjugants were observed earlier in animals dosed with this antibiotic. Molecular analysis of in vivo transconjugants containing the tet(M) gene showed that this gene was identical to tet(M) localized on the conjugative transposon Tn916. Primer-specific PCR confirmed that the Tn916 transposon was complete in all analyzed transconjugants and donors. This is the first study showing in vivo transfer of a Tn916-like antibiotic resistance transposon from L. lactis to E. faecalis. These data suggest that in certain cases food lactococci might be involved in the spread of antibiotic resistance genes to other lactic acid bacteria.

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“…Biofilms are considered to be hot spots for plasmid transfer. Several studies have found higher transfer frequencies under biofilm conditions than with planktonic cultures (Lampkowska et al ., ; Nguyen et al ., ; Hennequin et al ., ; Savage et al ., ). The plasmid used in this study, pB10, belongs to the incompatibility group IncP‐1β.…”

Section: Resultsmentioning

confidence: 99%

Biofilm models for the food industry: hot spots for plasmid transfer?

Meervenne

Weirdt

Coillie³

et al. 2014

Pathogens Disease

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Biofilms represent a substantial problem in the food industry, with food spoilage, equipment failure, and public health aspects to consider. Besides, biofilms may be a hot spot for plasmid transfer, by which antibiotic resistance can be disseminated to potential foodborne pathogens. This study investigated biomass and plasmid transfer in dual-species (Pseudomonas putida and Escherichia coli) biofilm models relevant to the food industry. Two different configurations (flow-through and drip-flow) and two different inoculation procedures (donor-recipient and recipient-donor) were tested. The drip-flow configuration integrated stainless steel coupons in the setup while the flow-through configuration included a glass flow cell and silicone tubing. The highest biomass density [10 log (cells cm-²)] was obtained in the silicone tubing when first the recipient strain was inoculated. High plasmid transfer ratios, up to 1/10 (transconjugants/total bacteria), were found. Depending on the order of inoculation, a difference in transfer efficiency between the biofilm models could be found. The ease by which the multiresistance plasmid was transferred highlights the importance of biofilms in the food industry as hot spots for the acquisition of multiresistance plasmids. This can impede the treatment of foodborne illnesses if pathogens acquire this multiresistance in or from the biofilm.

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A standardized conjugation protocol to asses antibiotic resistance transfer between lactococcal species

Cited by 47 publications

References 13 publications

Transfer of plasmid-mediated resistance to tetracycline in pathogenic bacteria from fish and aquaculture environments

Transfer of plasmid-mediated resistance to tetracycline in pathogenic bacteria from fish and aquaculture environments

Intra- and Interspecies Conjugal Transfer of Tn 916 -Like Elements from Lactococcus lactis In Vitro and In Vivo

Biofilm models for the food industry: hot spots for plasmid transfer?

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