Characterisation and antibiotic susceptibility profile of Clostridioides (Clostridium) difficile isolated from chicken carcasses

Gene	Primers	Amplicon size	Reference
tpi	F: 5′-AAAGAAGCTACTAAGGGTACAAA-3′	230 bp	15
R: 5′-CATAATATTGGGTCTATTCCTAC-3′
tcdA	F: 5′-AGATTCCTATATTTACATGACAATAT-3′	369 bp	15
R: 5′-GTATCAGGCATAAAGTAATATACTTT-3′
tcdB	F: 5′-GGAAAAGAGAATGGTTTTATTAA-3′	160 bp	15
R: 5′-ATCTTTAGTTATAACTTTGACATCTTT-3′
cdtA	F: 5′-TGAACCTGGAAAAGGTGATG-3′	353 bp	28
R: 5′-AGGATTATTTACTGGACCATTTG-3′
cdtB	F: 5′-CTTATTGCAAGTAAATACTGAGAGTACTATATC-3′	490 bp	28
R: 5′-ACCGGATCTCTTGCTTCAGTC-3′

Table 2

Susceptibility profiles of C. difficile isolates from chicken carcasses

n	Susceptibility	AMP (%)	AMC (%)	DA (%)	IMP (%)	MTZ (%)	TE (%)	VA (%)	CTX (%)
	Susceptible	48	69	39	6	61	66	67	2
		(69.6)	(100)	(56.5)	(8.7)	(88.4)	(95.7)	(97.1)	(2.9)
		19	0	14	1	0	1	0	0
69	Intermediate	(27.5)	(0)	(20.3)	(1.4)	(0)	(1.4)	(0)	(0)
		2	0	16	62	8	2	2	67
	Resistant	(2.9)	(0)	(23.2)	(89.9)	(11.6)	(2.9)	(2.9)	(97.1)

[i] AMP – ampicillin; AMC – amoxicillin-clavulanic acid; DA – clindamycin; IMP – imipenem; MTZ – metronidazole; TE – tetracycline; VA – vancomycin; CTX – cefotaxime; n – sample number

Table 3

The distribution of determined ribotypes in terms of antibiotic susceptibility

		AMP			AMC			DA			IMP			MTZ			TE			VA			CTX
RTs	n	S	I	R	S	I	R	S	I	R	S	I	R	S	I	R	S	I	R	S	I	R	S I	R
RT027	6	4	2	-	6	-	-	4	1	1	-	-	6	6	-	-	6	-	-	6	-	-	- -	6
RT087	4	3	1	-	4	-	-	3	1	-	-	-	4	4	-	-	4	-	-	4	-	-	- -	4
RT470	4	2	2	-	4	-	-	4	-	-	-	-	4	4	-	-	4	-	-	4	-	-	- -	4
RT085	4	4	-	-	4	-	-	3	1	-	-	-	4	4	-	-	4	-	-	4	-	-	- -	4
RT456	2	2	-	-	2	-	-	2	-	-	-	-	2	2	-	-	2	-	-	2	-	-	- -	2
RT020	2	2	-	-	2	-	-	1	1	-	-	-	2	2	-	-	2	-	-	2	-	-	- -	2
RT010	1	1	-	-	1	-	-	1	-	-	-	-	1	1	-	-	1	-	-	1	-	-	- -	1
RT003	1	1	-	-	1	-	-	1	-	-	-	-	1	1	-	-	1	-	-	1	-	-	- -	1
ML027	6	3	3	-	6	-	-	3	2	1	-	-	6	6	-	-	6	-	-	6	-	-	- -	6
NR	3	26	11	2	39	-	-	17	8	14	6	1	32	31	-	8	36	1	2	37	-	2	2 -	3
	9																							7
TOTAL	69	48	19	2	69	-	-	39	14	16	6	1	62	61	-	8	66	1	2	67	-	2	2 -	67

[i] AMP – ampicillin; AMC – amoxicillin-clavulanic acid; DA – clindamycin; IMP – imipenem; MTZ – metronidazole; TE – tetracycline; VA – vancomycin; CTX – cefotaxim; n – ribotype number; ML – most likely; S – susceptible; I – intermediate; R– resistant; RTs – ribotypes

Table 4

The distribution of the virulence genes and ribotypes of C. difficile isolates (n = 69)

Toxigenic genes			Samples (%)	Ribotypes (n^r)
tcdA⁺	tcdB⁺	cdtA/B⁺	17 (24.6)	027 (6), 003 (1), ML^#027 (6), NR^* (4)
tcdA⁺	tcdB⁺	cdtA/B⁻	14 (20.3)	087 (4), 020 (2), NR (8)
tcdA⁺	tcdB⁻	cdtA/B⁺	1 (1.5)	NR (1)
tcdA⁺	tcdB⁻	cdtA/B⁻	3 (4.4)	NR (3)
tcdA⁻	tcdB⁺	cdtA/B⁺	13 (18.8)	NR (13)
tcdA⁻	tcdB⁺	cdtA/B⁻	11 (15.9)	470 (4), 456 (2), NR (5)
tcdA⁻	tcdB⁻	cdtA/B⁺	0 (0)	ND^**
tcdA⁻	tcdB⁻	cdtA/B⁻	10 (14.5)	010 (1), 085 (4), NR (5)

[i] NR^* – new ribotype; ND^** – not detected; ML^# – most likely; n^r – number of ribotypes

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DOI: https://doi.org/10.2478/jvetres-2020-0052 | Journal eISSN: 2450-8608 (formerly 2300-3235)

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Language: English

Page range: 407 - 412

Submitted on: Jan 16, 2020

Accepted on: Jul 27, 2020

Published on: Aug 11, 2020

Published by: National Veterinary Research Institute in Pulawy

In partnership with: Paradigm Publishing Services

Keywords:

chicken,

C. difficile,

ribotype,

antibiotic susceptibility,

toxin

Related subjects:

Life sciences,

Molecular biology,

Microbiology and virology,

Life sciences, other,

Medicine,

Veterinary medicine

© 2020 Enver Baris Bingol, Hamparsun Hampikyan, Karlo Muratoglu, Esra Akkaya, Omer Cetin, Hilal Colak, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Volume 64 (2020): Issue 3 (September 2020)