Tetracycline and multidrug resistance in the oral microbiota: differences between healthy subjects and patients with periodontitis in Spain

Figures & data

Table 1. List of primers used in this study

Primers	Sequence 5ʹ-3’	Size (bp)	Tm	Tm Multiplex	Multiplex group	Ref.
tetMF tetMR	GCG TAC AAG CAC AGA CTC GT AGC CAT AGC GTA TCC CCT CC	1142	61	64	1	[6]
tetWF tetWR	GAG AGC CTG CTA TAT GCC AGC GGG CGT ATC CAC AAT GTT AAC	168	64			[57]
intF intR	GGC TAC AGA CCG AGT ACC AGC GGA ACT TGA CGT TCG CCA CT	684	61			[6]
ermBF ermBR	GGT AAA GGG CAT TTA ACG AC CGA TAT TCT CGA TTG ACC CA	494	55	60	2	[58]
tetQF tetQR	AGA ATC TGC TGT TTG CCA GTG CGG AGT GTC AAT GAT ATT GCA	167	50			[57]
tet32F tet32R	GAA CCA GAT GCT GCT CTT CAT AGC CAC GCC CAC ATG AT	620	57			[59]
tetLF tetLR	TCG TTA GCG TGC TGT CAT TC GTA TCC CAC CAA TGT AGC CG	267	55			[60]
tetOF tetOR	AAC TTA GGC ATT CTG GCT CAC TCC CAC TGT TCC ATA TCG TCA	515	55	58	3	[61]
tetSF tetSR	GAA AGC TTA CTA TAC AGT AGC AGG AGT ATC TAC AAT ATT TAC	168	50			[57]
tet31F tet31R	CAA TCA CGC CCA AAA GAA TGT GCC ATC CCA GTT TGT	564	53			[62]
tetBF tetBR	AAT AGC CAC TAA ATG GGG CG ATA ACA CCG GTT GCA TTG GT	243	58	56.5	4	[6]
tetKF tetKR	TCG ATA GGA ACA GCA GTA CAG CAG ATC CTA CTC CTT	169	52.8			[60]
tet37F tet37R	ATG GTT CGC TAT TAC TCT AAC ATC AGT CTC ATA TTT CGA CA	170	50			[6]
27 F 1544 R	GAG TTT GAT CCT GGC TCA G AGA AAG GAG GTG ATC CAG CC	approx. 1500	57	-	-	[63]

Table 2. Species identified from SPHS and SSP and the number of isolates carrying each antimicrobial resistance gene and their phenotypical resistance. The asterisk indicates significant differences between SPHS and SSP isolates. AMX: amoxicillin, CTX: cefotaxime, CHL: chloramphenicol, STR: streptomycin, ERY: erythromycin, KAN: kanamycin

		Antimicrobial resistance genes														Antimicrobial resistance
	Species	N	intTn	tet(W)	tet(O)	tet(M)	tet [31]	tet [32]	tet(B)	tet(L)	tet(S)	tet(Q)	tet [37]	tet(K)	erm(B)	AMX	CTX	CHL	STR	ERY	KAN
SSP	Actinomyces sp.	1	0	1	0	0	0	0	0	0	0	0	0	0	0	0	1	0	1	0	1
	Bacteroides sp.	2	0	0	0	0	0	0	0	0	0	2	0	0	0	0	2	0	2	2	2
	Gemella sp. *	2	2	0	0	2	0	0	0	0	0	0	0	0	0	1	0	0	1	2	2
	Granulicatella sp.	2	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Lachnoanaerobaculum sp.	3	2	0	0	0	0	0	0	0	0	0	0	0	2	1	2	0	2	2	2
	Mogibacterium sp.	1	0	1	0	0	0	1	0	0	0	0	0	0	0	0	1	0	0	1	1
	Neisseria sp.	2	0	0	0	0	0	0	0	0	0	0	0	0	0	0	2	0	0	2	0
	Peptoniphilus sp.	2	2	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Peptostreptococcus sp.	1	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Prevotella sp. *	48	16	0	0	3	0	18	7	0	0	27	0	2	3	28	36	2	15	24	42
	Proteus sp.	1	1	0	0	0	0	0	0	0	0	0	0	0	1	1	1	1	1	1	1
	Pseudomonas sp.	2	0	0	0	0	0	0	0	0	0	0	0	0	0	2	2	2	2	2	2
	Rothia sp.	2	1	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	1	2	2
	Slackia sp.	2	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	1	0
	Staphylococcus sp.	2	0	0	0	0	0	0	0	0	0	0	0	0	0	0	2	0	0	0	0
	Streptococcus sp.	276	248	0	18	116	0	31	0	12	0	3	0	0	128	20	56	29	48	188	271
	Veillonella sp.	6	6	0	0	5	0	0	0	0	0	0	0	0	2	1	4	0	4	5	2
	Not identified	10	2	0	0	2	0	1	1	0	0	1	0	0	1	1	2	1	1	3	4
SPHS	Abiotrophia sp.	5	5	0	0	2	0	0	0	0	0	0	0	0	4	0	0	1	0	5	2
	Bacilus sp.	1	1	0	0	1	0	0	0	0	0	0	0	0	1	0	0	0	0	1	1
	Butyrivibrio sp.	4	1	0	2	1	0	1	0	0	0	0	0	0	1	2	0	0	2	1	4
	Eubacterium sp.	3	2	1	0	3	0	1	1	0	0	0	0	0	2	0	0	0	1	3	3
	Gemella sp. *	13	13	1	0	8	0	0	1	0	0	1	0	0	0	1	1	4	0	5	4
	Granulicatella sp.	3	1	0	0	0	0	0	0	0	0	0	0	0	1	0	0	0	0	1	0
	Haemophilus sp.	4	2	1	0	2	0	1	2	0	0	0	0	0	3	0	0	0	0	4	1
	Neisseria sp.	2	0	0	0	2	0	0	0	0	0	0	0	0	2	0	0	0	1	2	0
	Prevotella sp. *	25	0	0	0	0	0	1	3	0	0	16	0	1	0	16	15	4	13	18	22
	Rothia sp.	4	1	3	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	1	2
	Staphylococcus sp.	5	1	0	0	0	0	0	0	0	0	0	0	3	4	3	3	2	3	4	0
	Streptococcus sp.	340	305	44	42	250	0	12	2	0	1	5	0	0	218	39	12	68	59	227	283
	Veillonella sp.	12	11	5	1	9	0	0	0	0	0	0	0	0	5	3	5	2	4	12	6
	Not identified	27	24	3	0	17	0	0	0	0	0	3	0	0	20	7	0	3	7	27	20

Table 3. Number of isolates resistant to other antimicrobials. The asterisk indicates significant differences between samples from periodontally healthy subjects (SPHS) and the samples from subjects with periodontitis (SSP) isolates. In parentheses, the percentage of isolates over the total number of isolates from each population. Define meaning of asterisk

Number of other antimicrobials	SPHS isolates (%)		SSP isolates (%)
0*	48	(10.71)		21	(5.75)
1	82	(18.30)		68	(18.63)
2	179	(39.96)		140	(38.36)
3	93	(20.76)		77	(21.10)
4	31	(6.92)		37	(10.14)
5	12	(2.68)		13	(3.56)
6*	3	(0.67)		9	(2.47)

Figure 1. Prevalence (%) of tet, int and erm(B) genes and the prevalence (%) of subgingival isolates resistant to different antimicrobials (KAN: kanamycin, ERY: erythromycin, STR: streptomycin, CHL: Chloramphenicol, AMX: Amoxicillin, CTX: cefotaxime. The asterisk indicates significant differences between the isolates obtained from samples from periodontally healthy subjects and from samples from subjects with periodontitis

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