Synergism effect of tunicamycin and amphotericin B causes suppression to the MP65 and ERG3 gene in oral associated-Candida albicans and C. dubliniensis

Figures & data

Figure 1. Effect of TM in combination with AmB on growth of Candida sp. Growth inhibition zones on MHA.

Table 1. Oligonucleotide sequences of MP65, ERG3 and ACT1 gene.

Gene target	Primer sequences (5′-3′)	Tm(°C) ^a
MP65		53.2
Forward	TGTTGTTGTCACTATTGGTAATGG
Reverse	CGGCAGCAGAAGAAGAAGC	54.9
ERG3		54.3
Forward	AACGTGCCACTACTGCCATT
Reverse	ACAGATGGCCAGTGTAACCA	52.4
ACT1		56.4
Forward	CGATAACGGTTCTGGTATG
Reverse	CCTTGATGTCTTGGTCTAC	55.1

^aMelting temperatures.

Table 2. Effect of TM and AmB alone and in combination against oral Candida spp. on agar diffusion assay.

	Antifungal activity (mm) of:
	Antifungal alone		Antifungal combination
Species	TM (500 µg/mL)	AmB (250 µg/mL)	TM (500 µg/mL) + AmB (250 µg/mL)
C. albicans	18	19.5	18
C. glabrata	19	19.5	21
C. parapsilosis	11	15	15
C. tropicalis	14	18	20
C. dubliniensis	19	19	22

Table 3. Susceptibility of planktonic Candida species to tunicamycin (TM) and amphotericin B (AmB) alone and in combination (MIC50).

	Planktonic cells
	Antifungal alone		Antifungal combination
	MIC50 (TM) (µg/mL)	MIC50 (AmB) (µg/mL)	MIC50 (TM + AmB) (µg/mL)	FIC index
C. albicans	15.6	7.8	7.8TM + 3.9AmB	1.0 = IND
C. glabrata	7.8	1.95	1.95TM + 0.98AmB	0.75 = P.SYN
C. parapsilosis	7.8	7.8	3.9TM + 1.95AmB	0.75 = P.SYN
C. tropicalis	15.6	3.91	3.9TM + 1.95AmB	0.75 = P.SYN
C. dubliniensis	7.8	3.9	3.9TM + 1.95AmB	0.5 = SYN

Note: MIC50 defined as the concentration of agent demonstrating 50% inhibition of the growth in comparison with that of the agent-free growth control.

IND = Indifference.

P.SYN = Partial synergistic.

SYN = Synergistic.

Figure 2. The expression level of MP65 and ERG3 gene in C. albicans and C. dubliniensis.