Activity of five antimicrobial peptides against periodontal as well as non-periodontal pathogenic strains

Figures & data

Table 1. Microbial strains used in this study for testing the activity of 5 antimicrobial peptides.

Species	Reference
Actinomyces israelii	ATCC 12102
Actinomyces johnsonii	clinical strain
Actinomyces naeslundii	ATCC 12104
Actinomyces odontolyticus	DSM 43331
Actinomyces oris	DSM 23056
Aggregatibacter actinomycetemcomitans	ATCC 29523
Atopobium parvulum	clinical strain
Bifidobacterium longum	clinical strain
Campylobacter concisus	clinical strain
Campylobacter rectus	DSM 3260
Dialister pneumosintes	DSM 11619
Eikenella corrodens	ATCC 23834
Enterobacter cloacae	clinical strain
Enterococcus faecalis	ATCC 29212
Escherichia coli	ATCC 25922
Fusobacterium nucleatum ssp. fusiforme	ATCC 51190
Fusobacterium nucleatum ssp. nucleatum	ATCC 25586
Fusobacterium nucleatum ssp. polymorphum	ATCC 10953
Fusobacterium nucleatum ssp. vincentii	ATCC 49256
Megasphaera sp.	DSM 102144
Parvimonas micra	clinical strain
Porphyromonas gingivalis	clinical strain
Prevotella intermedia	DSM 20706
Prevotella loescheii	clinical strain
Prevotella melaninogenica	clinical strain
Prevotella nigrescens	DSM 13386
Pseudomonas aeruginosa	ATCC 27853
Selenomonas flueggei	clinical strain
Selenomonas sputigena	clinical strain
Slackia exigua	clinical strain
Staphylococcus aureus	ATCC 29213
Streptococcus anginosus	DSM 20563
Streptococcus constellatus ssp. constellatus	DSM 20575
Streptococcus mutans	DSM 20523
Veillonella parvula	clinical strain
Candida albicans	DSM 11949

Table 2. Growth categories of different levels of reduced growth of the bacteria on the agar peptide plates.

Growth category	Growth on agar plates
Complete inhibition	no colonies
Significant growth reduction	few or sparse countable colonies
Slight growth reduction	confluent growth with declining tendency
No growth reduction	confluent growth comparable to growth control

Table 3. Bacterial growth on agar plates with different concentrations of the five AMPs cultivated in anaerobic atmosphere. Concentrations between 0.125 µg/ml and 8 µg/ml of all peptides were tested. In addition, nisin concentrations up to 128 µg/ml were tested.

Species	Peptide [µg/ml]	0,125	0,25	0,5	1	2	4	8	16	32	64	128
Megasphaera sp.	Nisin	+	+	+	±	-	-	-	-	-	-	-
	Lactof	+	+	+	+	+	+	+
	Melit	+	+	+	+	+	+	+
	Para-1	+	+	+	+	+	+	+
	LL-37	+	+	+	+	+	+	+
P. micra	Nisin	+	+	+	+	+	+	±	-	-	-	-
	Lactof	+	+	+	+	+	+	+
	Melit	+	+	+	+	+	+	+
	Para-1	+	+	+	+	+	+	+
	LL-37	+	+	+	+	+	+	+
B. longum	Nisin	+	+	+	±	-	-	-	-	-	-	-
	Lactof	+	+	+	+	+	+	+
	Melit	+	+	+	+	+	+	+
	Para-1	+	+	+	+	+	+	+
	LL-37	+	+	+	+	+	+	+
A. israelii	Nisin	+	+	+	+	+	+	+	-	-	-	-
	Lactof	+	+	+	+	+	+	+
	Melit	+	+	+	+	+	+	+
	Para-1	+	+	+	+	+	+	+
	LL-37	+	+	+	+	+	+	+
A. naeslundii	Nisin	+	+	+	+	+	+	+	+	-	-	-
	Lactof	+	+	+	+	+	+	+
	Melit	+	+	+	+	+	+	+
	Para-1	+	+	+	+	+	+	+
	LL-37	+	+	+	+	+	+	+
A. odontolyticus	Nisin	+	+	+	+	+	+	+	-	-	-	-
	Lactof	+	+	+	+	+	+	+
	Melit	+	+	+	+	+	+	+
	Para-1	+	+	+	+	+	+	+
	LL-37	+	+	+	+	+	+	+
S. aureus	Nisin	+	+	+	+	+	+	+	+	+	+	-
	Lactof	+	+	+	+	+	+	+
	Melit	+	+	+	+	+	+	+
	Para-1	+	+	+	+	+	+	+
	LL-37	+	+	+	+	+	+	+
S. anginosus	Nisin	+	+	+	+	+	+	+	+	+	+	-
	Lactof	+	+	+	+	+	+	+
	Melit	+	+	+	+	+	+	+
	Para-1	+	+	+	+	+	+	+
	LL-37	+	+	+	+	+	+	+
S. constellatus ssp. con.	Nisin	+	+	+	+	+	+	+	+	+	-	-
	Lactof	+	+	+	+	+	+	+
	Melit	+	+	+	+	+	+	+
	Para-1	+	+	+	+	+	+	+
	LL-37	+	+	+	+	+	+	+
P. intermedia	Nisin	+	+	+	+	+	+	+	+	+	-	-
	Lactof	+	+	+	+	+	+	+
	Melit	+	+	+	+	+	+	+
	Para-1	+	+	+	+	+	+	+
	LL-37	+	+	+	+	+	+	+
A. johnsonii, A. oris, A. actinomycetemcomitans, A. parvulum, C. concisus, C. rectus, D. pneumosintes, E. corrodens, E. cloacae, E. faecalis, E. coli, F nucleatum (fusiforme, nucleatum, polymorphum, vincentii), P. gingivalis, P. loescheii, P. melaninogenica, P. nigrescens, S. flueggei, S. sputigena, S. exigua, S. mutans, V. parvula, C. albicans	Nisin	+	+	+	+	+	+	+	+	+	+	+
	Lactof	+	+	+	+	+	+	+
	Melit	+	+	+	+	+	+	+
	Para-1	+	+	+	+	+	+	+
	LL-37	+	+	+	+	+	+	+

‘+’ – growth categories ii, iii, iv; ‘-’ – growth category i; ‘±’ – growth category i on at least one agar plate in at least one experiment; ‘empty table element’ – not measured; ‘Lactof’ – Lactoferrin; ‘Melit’ – Melittin; ‘Para-1’ – Parasin-1

P. aeruginosa was not cultivated anaerobically.

Table 4. Bacterial growth on agar plates with different concentrations of nisin cultivated under aerobic atmosphere. In test 1 and 2 nisin concentrations between 0.125 µg/ml and 8 µg/ml were tested. In test 3 and 4 nisin concentrations between 0.5 µg/ml and 128 µg/ml were tested. (Bacteria with no growth inhibition are not shown.).

Nisin [µg/ml] aerobic atmosphere		0.125	0.25	0.5	1	2	4	8	16	32	64	128
A. israelii	Test 1	+	+	+	+	+	+	+
	Test 2	+	+	+	+	+	+	+
	Test 3			+	+	+	+	+	+	-	-	-
	Test 4			+	+	+	+	+	+	-	-	-
A. naeslundii	Test 1	+	+	+	+	+	+	+
	Test 2	+	+	+	+	+	+	+
	Test 3			+	+	+	+	+	+	-	-	-
	Test 4			+	+	+	+	+	+	+	-	-
A. odontolyticus	Test 1	+	+	+	+	+	+	+
	Test 2	+	+	+	+	+	+	+
	Test 3			+	+	+	+	+	-	-	-	-
	Test 4			+	+	+	+	+	-	-	-	-
S. aureus	Test 1	+	+	+	+	+	+	+
	Test 2	+	+	+	+	+	+	+
	Test 3			+	+	+	+	+	+	+	-	-
	Test 4			+	+	+	+	+	+	+	-	-
S. anginosus	Test 1	+	+	+	+	+	+	+
	Test 2	+	+	+	+	+	+	+
	Test 3			+	+	+	+	+	+	+	-	-
	Test 4			+	+	+	+	+	+	+	+	-
S. constellatus ssp. con.	Test 1	+	+	+	+	+	+	+
	Test 2	+	+	+	+	+	+	+
	Test 3			+	+	+	+	+	+	+	-	-
	Test 4			+	+	+	+	+	+	+	-	-

‘+’ – growth categories ii, iii, iv; ‘-’ – growth category i; ‘empty table element’ – not measured.

Table 5. Antimicrobial activity of nisin, melittin and lactoferrin tested by the agar dilution method. Reduced growth of bacteria on the agar peptide plates was compared with the growth control plates and differentiated into different levels (i, ii, iii) of reduced growth. (Bacteria with no growth reduction (iv) are not shown.).

Peptide	Strain	i) complete inhibition, MIC in [µg/ml]	ii) significant growth reduction, concentration in [µg/ml]	iii) slight growth reduction, concentration in [µg/ml]
Nisin	A. israelii	16, 32
	A. naeslundii	32, 64
	A. odontolyticus	16
	A. parvulum		128
	B. longum	1, 2
	Megasphaera sp.	1, 2
	P. micra	8, > 8
	P. gingivalis			128
	P. intermedia	64
	P. loescheii		128
	P. nigrescens		128
	S. flueggei			128
	S. exigua		128
	S. anginosus	64, 128
	S. aureus	64, 128
	S. constellatus ssp. con.	64
	V. parvula		128
Melittin	B. longum		8
	Megasphaera sp.		8
	P. micra		8
Lactoferrin	Megasphaera sp.		8
	P. micra			8
	S. flueggei			8