European Food Research and Technology
Safety assessment, genetic relatedness and bacteriocin activity of potential probiotic Lactococcus lactis strains from rainbow trout (Oncorhynchus mykiss, Walbaum) and rearing environment Carlos Araújoa-e, Estefanía Muñoz-Atienzaa, Martha Ramíreza, Patrícia Poetab-c, Gilberto Igrejasd-e, Pablo E. Hernándeza, Carmen Herranza and Luis M. Cintasa* a Grupo de Seguridad y Calidad de los Alimentos por Bacterias Lácticas, Bacteriocinas y Probióticos (Grupo SEGABALBP), Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040-Madrid, Spain
b Centre for Animal Science and Veterinary, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal
c Veterinary Science Department, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal
d Institute for Biotechnology and Bioengineering, Centre of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal
e Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal * Corresponding author. Phone: +34913943751. Fax: +34913943743. E-mail address: lcintas@vet.ucm.es (L.M. Cintas). Online Resource 1 Table S1 Oligonucleotide primers used in this study Table S1 Oligonucleotide primers used in this study
Target gene
| Primer
| 5’–3’ sequencea
| PCR fragment size (bp)
| Annealing temperature (°C)
| Reference
| aad(E)
| aadEI
| GCAGAACAGGATGAACGTATTCG
| 369
| 55
| [83]
| aadEII
| ATCAGTCGGAACTATGTCCC
| aph(3′)-IIIa
| aph(3')-F
| GCCGATGTGGATTGCGAAAA
| 292
| 60
| [87]
| aph(3')-R
| GCTTGATCCCCAGTAAGTCA
| lin(A)
| linA
| GGTGGCTGGGGGGTAGATGTATTAACTGG
| 323
| 57
| [85]
| linA'
| GCTTCTTTTGAAATACATGGTATTTTTCGATC
| lin(B)
| LINB1
| CCTACCTATTGTTTGTGGAA
| 925
| 54
| [80]
| LINB2
| ATAACGTTACTCTCCTATTC
| tetB(P)
| TetB/P-FW
| AAAACTTATTATATTATAGTG
| 169
| 46
| [83]
| TetB/P-RV
| TGGAGTATCAATAATATTCAC
| tet(K)
| tetKI
| CAATACCTACGATATCTA
| 352
| 50
| [83]
| tetKII
| TTGAGCTGTCTTGGTTCA
| tet(L)
| tetLI
| TGGTCCTATCTTCTACTCATTC
| 385
| 53
| [83]
| tetLII
| TTCCGATTTCGGCAGTAC
| tet(M)
| tetMI
| GGTGAACATCATAGACACGC
| 401
| 45
| [83]
| tetMII
| CTTGTTCGAGTTCCAATGC
| tet(O)
| tetOI
| AGCGTCAAAGGGGAATCACTATCC
| 1,723
| 55
| [83]
| tetOII
| CGGCGGGGTTGGCAAATA
| tet(Q)
| TetQ-FW
| AGAATCTGCTGTTTGCCAGTG
| 169
| 63
| [83]
| TetQ-RV
| CGGAGTGTCAATGATATTGCA
| tet(S)
| tetS-FW
| ATCAAGATATTAAGGAC
| 573
| 55
| [83]
| tetS-RV
| TTCTCTATGTGGTAATC
| tet(T)
| TetT-FW
| AAGGTTTATTATATAAAAGTG
| 169
| 46
| [83]
| TetT-RV
| AGGTGTATCTATGATATTTAC
| tet(W)
| tetWI
| GGMCAYRTGGATTTYWTIGC
| 1,187
| 64
| [83]
| tetWII
| TCIGMIGGIGTRCTIRCIGGRC
| van(A)
| vanA-36F
| TTGCTCAGAGGAGCATGACG
| 957
| 65
| [9]
| vanA-992R
| TCGGGAAGTGCAATACCTGC
| van(B)
| vanB-23F
| TTATCTTCGGCGGTTGCTCG
| 994
| 62
| [9]
| vanB-1016R
| GCCAATGTAATCAGGCTGTC
| hdc
| CL1
| CCWGGWAAWATWGGWAATGGWTA
| 500
| 48
| [84]
| JV17HC
| AGACCATACACCATAACCTT
| ldc
| CAD2-F
| CAYRTNCCNGGNCAYAA
| 1,185
| 53
| [82]
| CAS2-R
| GGDATNCCNGGNGGRTA
| odc
| 3
| GTNTTYAAYGCNGAYAARACNTAYTTYGT
| 1,446
| 52
| [86]
| 16
| TACRCARAATACTCCNGGNGGRTANGG
| tdc
| TD5
| CAAATGGAAGAAGAAGTAGG
| 1,100
| 48
| [81]
| TD2
| ACATAGTCAACCATRTTGAA
| ERIC-PCR
| ERIC-1R
| ATGTAAGCTCCTGGGGATTCAC
| variable
| 46
| [33]
| ERIC-2
| AAGTAAGTGACTGGGGTGAGCG
| rep-PCR
| (GTG)5
| GTGGTGGTGGTGGTG
| variable
| 40
| [36]
| a K=G or T; R=A or G; W=A or T; Y=C or T; S=C or G; M=A or C; D=A, G, or T; N=A, G, C, or T |