Genotypic evaluation of antimicrobial resistance in Staphylococcus spp. isolated from bovine clinical mastitis

Benites, N.R.; Hora, A.S.; Mello, P.L.; Laes, M.A.; Brandão, P.E.; Melville, P.A.; Zuniga, E.

doi:10.1590/1678-4162-11984

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Veterinary Medicine • Arq. Bras. Med. Vet. Zootec. 73 (02) • Mar-Apr 2021 • https://doi.org/10.1590/1678-4162-11984 copy

Genotypic evaluation of antimicrobial resistance in Staphylococcus spp. isolated from bovine clinical mastitis

[Avaliação genotípica da resistência antimicrobiana em Staphylococcus spp. Isolados de mastite clínica bovina]

Authorship SCIMAGO INSTITUTIONS RANKINGS

ABSTRACT

Bovine clinical mastitis caused by Staphylococcus spp. is a serious and widespread disease in the world of dairy farming. Antimicrobial therapy is of fundamental importance in the prevention and treatment of infectious mastitis, but the indiscriminate use of antimicrobials acts as a determining factor for the spread of the disease. The present study evaluated the resistance profiles of 57 Staphylococcus spp. isolated from bovine clinical mastitis to beta-lactams and gentamicin, relating characteristics of phenotype (in vitro susceptibility tests) and genotype (detection and expression of genes encoding resistance - mecA, mecA_LGA251, blaZ, femA, femB, and aacA-aphD - using PCR and RT-PCR, respectively). One or more genes coding for resistance to different antimicrobials were detected in 50 Staphylococcus spp. isolates. The femA and femB genes were the most frequent (75.4% for both). The observed expression of the genes was as follows: blaZ (60%), femA (39.5%), aacA-aphD (50%), femB (32.6%), mecA (8.3%), and mecA_LGA251 (0%). Considering the relevance of the genus Staphylococcus to bovine mastitis, this study aimed to elucidate aspects regarding the genotypic and phenotypic profiles of these microorganisms so as to contribute to the development of effective strategies for mastitis control.

Keywords:
staphylococcal mastitis; antimicrobial resistance genes; gene expression

Antimicrobial	MIC (Range for most strains - µg/mL)	MIC₅₀	MIC₉₀
Oxacillin (N = 4)	0.5-2	≤ 0.5	≤ 1
Penicillin (N=24)	0.25-16	≤ 2	≤ 16
Amoxicillin (N=24)	0.5-2	≤ 1	≤ 2
Ampicillin (N=24)	0.5-2	≤ 1	≤ 2
Gentamicin (N=5)	16-128	≤ 64	≤ 128

Coagulase	Staphylococcus Species	N	femA				femB				mecA				aacA-aphD				mecA _LGA251				blaZ
Coagulase	Staphylococcus Species	N	detection		expression		detection		expression		detection		expression		detection		expression		detection		expression		detection		expression
			N	%	N	%	N	%	N	%	N	%	N	%	N	%	N	%	N	%	N	%	N	%	N	%
Positive	S. aureus	41	40	97.6	17	42.5	40	97.6	14	35	7	17.1	0	0	9	22.0	5	55.6	0	0.0	0	0	1	2.4	1	100

other CPS	S. hyicus	6	0	0.0	0	0	0	0.0	0	0	3	50.0	0	0	2	33.3	0	0	0	0.0	0	0	1	16.7	0	0
	S. agnetis	2	2	100.0	0	0	2	100.0	0	0	1	50.0	0	0	2	100.0	0	0	0	0.0	0	0	0	0.0	0	0
	CPS total	8	2	25.0	0	0	2	25.0	0	0	4	50.0	0	0	4	50.0	0	0	0	0.0	0	0	1	12.5	0	0

Negative CNS	S. xylosus	1	1	100.0	0	0	1	100.0	0	0	0	0.0	0	0	0	0.0	0	0	0	0.0	0	0	0	0.0	0	0
	S.epidermidis	2	0	0.0	0	0	0	0.0	0	0	0	0.0	0	0	0	0.0	0	0	0	0.0	0	0	0	0.0	0	0
	S. equorum	2	0	0.0	0	0	0	0.0	0	0	0	0.0	0	0	0	0.0	0	0	1	50.0	0	0	1	50.0	0	0
	S.haemolyticus	3	0	0.0	0	0	0	0.0	0	0	1	33.3	1	100	3	100.0	3	100	0	0.0	0	0	2	66.7	2	100
	CNS total	8	1	12.5	0	0	1	12.5	0	0	1	12.5	1	100	3	37.5	3	100	1	12.5	0	0	3	37.5	2	66.7
	Total	57	43	75.4	17	39.5	43	75.4	14	32.6	12	21.1	1	8.33	16	28.1	8	50	1	1.8	0	0	5	8.8	3	60

gene	Gene Detection	N	%	Gene Expression	N	%	Ampicillin/ Amoxicillin		Oxacillin		Penicillin		Gentamicin
gene	Gene Detection	N	%	Gene Expression	N	%	N	%	N	%	N	%	N	%
femA	present	43	75.4	present	17	39.5	6	35.3	0	0.0	6	35.3	3	17.6
	present	43	75.4	absent	26	60.5	11	42.3	1	3.8	11	42.3	0	0.0
	absent	14	24.6	--	--	--	7	50.0	3	21.4	7	50.0	2	14.3
femB	present	43	75.4	present	14	32.6	4	28.6	0	0.0	4	28.6	3	21.4
	present	43	75.4	absent	29	67.4	13	44.8	1	3.4	13	44.8	0	0.0
	absent	14	24.6	--	--	--	7	50.0	3	21.4	7	50.0	2	14.3
mecA	present	12	21.1	present	1	8.3	1	100.0	1	100.0	1	100.0	1	100.0
	present	12	21.1	absent	11	91.7	3	27.3	0	0.0	3	27.3	0	0.0
	absent	45	78.9	--	--	--	20	44.4	3	6.7	20	44.4	4	8.9
mecA_LGA251	present	1	1.8	present	0	0.0	0	0.0	0	0.0	0	0.0	0	0.0
	present	1	1.8	absent	1	100.0	0	0.0	0	0.0	0	0.0	0	0.0
	absent	56	98.2	--	--	--	24	42.9	4	7.1	24	42.9	5	8.9
blaZ	present	5	8.8	present	3	60.0	3	100.0	1	33.3	3	100.0	1	33.3
	present	5	8.8	absent	2	40.0	0	0.0	0	0.0	0	0.0	0	0.0
	absent	52	91.2	--	--	--	21	40.4	3	5.8	21	40.4	4	7.7
aacA-aphD	present	16	28.1	present	8	50.0	3	37.5	1	12.5	3	37.5	5	62.5
	present	16	28.1	absent	8	50.0	0	0.0	0	0.0	0	0.0	0	0.0
	absent	41	71.9	--	--	--	21	51.2	3	7.3	21	51.2	0	0.0

Universidade Federal de Minas Gerais, Escola de Veterinária Caixa Postal 567, 30123-970 Belo Horizonte MG - Brazil, Tel.: (55 31) 3409-2041, Tel.: (55 31) 3409-2042 - Belo Horizonte - MG - Brazil
E-mail: abmvz.artigo@gmail.com

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[1] *Autor para correspondência (corresponding author) E-mail: eveline.zuniga@usp.br