RESUMO
Staphylococcus spp. is one of the pathogens that cause bovine mastitis and may present multiple resistance to different antimicrobial groups. The aim of this study was to phenotypically identify Staphylococcus spp. isolates obtained from bovine milk and to characterize their antimicrobial resistance profile. The 101 strains were classified by phenotypic tests, their resistance to oxacillin, cefoxitin, penicillin, ampicillin, tetracycline, kanamycin, sulfamethoxazole / trimethoprim, clindamycin and erythromycin was determined by the Kirby-Bauer technique and the presence of resistance genes by PCR. A total of 65 strains was S. aureus and 36 strains were coagulase-negative staphylococci (CoNS). We found different patterns of resistance to antibiotics evaluated in strains of S. aureus and CoNS, only the resistance to ampicillin was found associated with the species (p<0.005). In the 101 strains, the mecA gene was detected in 27%, aph(3')-IIIa in 75.2%, aac(6')/aph(2")-3 in 47.4%, ant(4')-Ia in 32.7%, tetM in 63% and tetK in 43.6%; however, no association was found with the resistance to penicillin, ampicillin, cefoxitin, kanamycin and tetracycline, respectively (p>0.05). On the other hand, the blaZ gene was found in 59.4% of the 101 strains and the ermCgene in 62.3%, which was associated with resistance to ß-lactams and macrolides, respectively (p<0.001). In this study, antimicrobial multiresistance was found in S. aureus and CoNS strains. This finding impacts on the dairy industry, representing a risk to public health.
Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Leite/microbiologia , Staphylococcus/efeitos dos fármacos , Staphylococcus/isolamento & purificação , Animais , Bovinos , Colômbia , Indústria de Laticínios , Feminino , Fenótipo , Staphylococcus/classificação , Staphylococcus/genéticaRESUMO
The objective of this study was to identify twelve Brucella abortus isolates of bovine origin from the department of Nariño in Colombia up to the biovar level. These isolates are included in the collection of the Germplasm Bank of Microorganisms of Animal Health Interest - Bacteria and Virus (BGSA-BV). The identification was carried out through conventional methods such as macro and microscopic morphological descriptions, enzymatic activity, biochemical profile, substrate use and sensitivity to dyes. Complementary genotypic characterization was carried out using multiplex PCR for B. abortus, Brucella melitensis, Brucella ovis, and Brucella suis-Erytritol (AMOS-ERY-PCR), RFLP-IS711, by southern blot hybridization, as well as by the multiple locus variable number of tandem repeat analysis (MLVA) using the ery gene and the insertion sequence IS711 and variable number of tandem repeats (VNTR) as molecular markers. The results of the phenotypic and molecular characterization allowed to identify twelve isolates as B. abortus biovar 4 as well as to differentiate field from vaccine strains. This is the first study on the phenotypic and molecular identification of B. abortus isolates in Colombia. It was concluded that the phenotypic and molecular identification of twelve isolates as B. abortus biovar 4 could be achieved using conventional and molecular techniques with enough resolution power. The identification of these isolates to the biovar level in taxonomic and epidemiological terms will allow the use of this genetic resource as reference strains in future research. This finding constitutes the basis for identifying biotypes not previously reported in the country that might be useful to support brucellosis survey programs in Colombia.
Assuntos
Brucella abortus/isolamento & purificação , Brucelose Bovina/microbiologia , Animais , Técnicas Bacteriológicas , Bancos de Espécimes Biológicos , Biomarcadores , Brucella abortus/classificação , Brucella abortus/genética , Brucella abortus/ultraestrutura , Brucelose Bovina/epidemiologia , Bovinos , Colômbia/epidemiologia , DNA Bacteriano/genética , Genes Bacterianos , Genótipo , Repetições Minissatélites , Reação em Cadeia da Polimerase Multiplex , FenótipoRESUMO
The objective of this study was to identify twelve Brucella abortus isolates of bovine origin from the department of Nariño in Colombia up to the biovar level. These isolates are included in the collection of the Germplasm Bank of Microorganisms of Animal Health Interest -Bacteria and Virus (BGSA-BV). The identification was carried out through conventional methods such as macro and microscopic morphological descriptions, enzymatic activity, biochemical profile, substrate use and sensitivity to dyes. Complementary genotypic characterization was carried out using multiplex PCR for B. abortus, Brucella melitensis, Brucella ovis, and Brucella suis-Erytritol (AMOS-ERY-PCR), RFLP-IS711, by southern blot hybridization, as well as by the multiple locus variable number of tandem repeat analysis (MLVA) using the ery gene and the insertion sequence IS711 and variable number of tandem repeats (VNTR) as molecular markers. The results of the phenotypic and molecular characterization allowed to identify twelve isolates as B. abortus biovar 4 as well as to differentiate field from vaccine strains. This is the first study on the phenotypic and molecular identification of B. abortus isolates in Colombia. It was concluded that the phenotypic and molecular identification of twelve isolates as B. abortus biovar 4 could be achieved using conventional and molecular techniques with enough resolution power. The identification of these isolates to the biovar level in taxonomic and epidemiological terms will allow the use of this genetic resource as reference strains in future research. This finding constitutes the basis for identifying biotypes not previously reported in the country that might be useful to support brucellosis survey programs in Colombia.
El objetivo de este estudio fue identificar 12 aislamientos de Brucella abortus de origen bovino procedentes del departamento de Narino, Colombia, hasta la descripción de biovar. Estos aislamientos conforman la colección del Banco de Germoplasma de Microorganismos de Interés en Salud Animal, Bacterias y Virus. La identificación se hizo mediante métodos convencionales, como la descripción morfológica macro y microscópica de actividad enzimática, de perfiles bioquímicos, de utilización de sustratos y de sensibilidad a colorantes. Se hizo una caracterización genotipica complementaria mediante PCR múltiple para Brucella abortus, Brucella melitensis, Brucella ovisy Brucella suis-eritritol (AMOS-ERY-PCR); RFLP-/S7II; hibridación Southern blot y análisis multi-locus de repeticiones en tándem de número variable (MLVA), empleando como marcadores moleculares el gen ery, la secuencia de inserción /S711 y el número variable de repeticiones en tándem (VNTR). Los resultados de la caracterización fenotípica y molecular permitieron identificar 12 aislamientos de campo como B. abortus biovar 4 y diferenciar cepas de campo de cepas vacunales. Este es el primer estudio de identificación fenotípica y molecular de aislamientos de B. abortus en Colombia. Por su importancia taxonómica y epidemiológica, la identificación de estos aislamientos hasta el nivel de biovar permitirá disponer de recursos genéticos que se pueden emplear como cepas de referencia en futuras investigaciones. Estos resultados pueden considerarse como una base para la identificación de biotipos no reportados en el país y podrán ser utilizados en programas de monitoreo y vigilancia de la brucelosis bovina en Colombia.