Sistema microbiológico de cinco bioensayos para la detección de residuos de antibióticos / Five-assay microbiological system for the screeningf antibiotic residues

Abstract A novel microbiological system in microtiter plates consisting of five bioassays is presented for the detection and classification of antibiotic residues in milk. The bioassays were optimized for the detection of beta-lactams (Bioassay B: Geobacillus stearothermophilus), macrolides (Bioassay M: Bacillus megaterium with fusidic acid), tetracyclines (Bioassay T: B. megaterium with chloramphenicol), quinolones (Bioassay Q: Bacillus licheniformis) and sulfamides (Bioassay QS: B. licheniformis with trimethoprim) at levels near the maximum residue limits (MRL). The response of each bioassay was interpreted visually (positive or negative) after 4-5.5h of incubation. The system detects and classifies beta-lactams (5 pg/l of amoxicillin, 4 pg/l of ampicillin, 36 pg/l of cloxacillin, 22 pg/l of amoxicillin, 3 pg/l of penicillin, 114 pg/l of cephalexin, 89pg/l of cefoperazone and 116 pg/l of ceftiofur), tetracyclines (98 pg/l of chlortetracycline, 92 pg/l of oxytetracycline and 88 pg/l of tetracycline), macrolides (33 pg/l of erythromycin, 44 pg/l of tilmicosin and 50 pg/l of tylosin), sulfonamides (76 pg/l of sulfadiazine, 85 pg/l of sulfadimethoxine, 77 pg/l of sulfamethoxazole and 87pg/l of sulfathiazole) and quinolones (94 pg/l of ciprofloxacin, 98 pg/l of enrofloxacin and 79 pg/l marbofloxacin). In addition, the specificity values were high for B, T, Q (99.4%), M (98.8%) and QS (98.1%) bioassays. The control of antibiotics through this system can contribute to improving the quality and safety of dairy products.

Resumen Se presenta un novedoso sistema microbiológico en placas de microtitulación compuesto por 5 bioensayos para la detección y clasificación de residuos de antibióticos en leche. Los bioensayos fueron optimizados para la detección de betalactámicos (bioensayo B: Geobacillus stearothermophilus), macrólidos (bioensayo M: Bacillus megaterium con ácido fusídico), tetraciclinas (bioensayo T: Bacillus megaterium con cloranfenicol), quinolonas (bioensayo Q: Bacillus licheniformis) y sulfamidas (bioensayo QS: Bacillus licheniformis con trimetoprima), a niveles cercanos a los límites máximos de residuos (LMR). La respuesta de cada bioensayo se interpretó visualmente (positiva o negativa) después de 4 a 5,5 h de incubación. El sistema detecta y clasifica betalactámicos (5 pg/l de amoxicilina, 4 pg/l de ampicilina, 36 pg/l de cloxacilina, 22 pg/l de amoxicilina, 3 pg/l de penicilina, 114 pg/l de cefalexina, 89 pg/l de cefoperazona y 116 pg/l de ceftiofur), tetraciclinas (98 pg/l de clortetraciclina, 92 pg/l de oxitetraciclina y 88 pg/l de tetraciclina), macrólidos (33 pg/l de eritromicina, 44 pg/l de tilmi-cosina y 50 pg/l de tilosina), sulfamidas (76 pg/l de sulfadiacina, 85 pg/l de sulfadimetoxina, 77 pg/l de sulfametoxazol y 87 pg/l de sulfatiazol) y quinolonas (94 pg/l de ciprofloxacina, 98 pg/l de enrofloxacina y 79pg/l de marbofloxacina). Además, los valores de especificidad fueron altos para los bioensayos B, T, Q (99,4%), M (98,8%) y QS (98,1%). El control de residuos de antibióticos mediante este sistema puede contribuir a mejorar la calidad e inocuidad de los productos lácteos.

Detección de residuos de quinolonas en carne bovina de venta en mercados municipales de la ciudad de Guatemala / Quinolone antibiotic residues in beef meat sold in municipal markets of Guatemala City

La resistencia bacteriana es una amenaza creciente para la salud pública mundial. El mal uso y uso excesivo de antimicrobianos en personas y animales está acelerando este proceso. En el caso de residuos antimicrobianos en productos de origen animal, se sabe que producen reacciones alérgicas, así como inducción de resistencia en bacterias transmitidas por alimentos como Salmonella, Campylobacter y Escherichia coli. Este estudio tuvo como objetivo detectar la presencia de residuos de quinolonas en carne bovina de venta en mercados municipales de la ciudad de Guatemala. Se obtuvieron 161 muestras para análisis. La detección de quinolonas se realizó por metodología de Inmunoensayo Asociado a Enzima (Elisa). Siete de 161 (5%) contenían residuos de quinolonas. Los niveles medios (desviación estándar) detectados fueron 16.497(1.69) µg/kg; ningún límite fue superior al límite máximo residual permitido (100 µg/kg). Este estudio indicó que existen residuos de antibiótico en algunas carnes de res en Guatemala.

Antimicrobial resistance is an increasingly serious threat to global public health. The misuse and overuse of antimicrobials in people and animals is an accelerating factor for this process. In the case of antimicrobial residues in animal origin products, it is known that they cause allergic reactions as well as induction of resistance in bacteria such as Salmonella, Campylobacter and Escherichia coli. This study aimed to find the presence of quinolones residues in beef meat. A total of 161 samples were collected from municipal markets in Guatemala City for analysis. Extraction and determination of quinolones was performed by Enzyme­linked Immunoassay (Elisa) methodology. Seven of 161 (5%) contained quinolone residues. The mean levels (±SE) of quinolones were 16.497(1.69) µg/kg; no limit was superior than the maximum residual limit allowed (100 µg/kg) by FDA. Even though none of the samples exceeded the upper limit it is important that some residues of quinolone were found.

Evaluation of the influence of fluoroquinolone chemical structure on stability: forced degradation and in silico studies

ABSTRACT Fluoroquinolones are a known antibacterial class commonly used around the world. These compounds present relative stability and they may show some adverse effects according their distinct chemical structures. The chemical hydrolysis of five fluoroquinolones was studied using alkaline and photolytic degradation aiming to observe the differences in molecular reactivity. DFT/B3LYP-6.31G* was used to assist with understanding the chemical structure degradation. Gemifloxacin underwent degradation in alkaline medium. Gemifloxacin and danofloxacin showed more degradation perceptual indices in comparison with ciprofloxacin, enrofloxacin and norfloxacin in photolytic conditions. Some structural features were observed which may influence degradation, such as the presence of five member rings attached to the quinolone ring and the electrostatic positive charges, showed in maps of potential electrostatic charges. These measurements may be used in the design of effective and more stable fluoroquinolones as well as the investigation of degradation products from stress stability assays.

Efficacy evaluation of some antibiotics against syrian brucella spp isolates, in vitro

Brucellosis is an endemic zoonosis in Syria, affecting large numbers of animals and there are an increasing number of cases in humans. The aim of this study is to investigate the in vitro efficacy of various traditional and new antibiotics against89 Brucella isolates (isolated from domestic animals) collected from different Syrian regions. Minimum inhibitory concentrations (MICs) of seventeen antibiotics were determined. Ciprofloxacin and ofloxacin were the most effective antibiotics, whereas sparfloxacin, levofloxacin, doxycycline and tetracycline had a moderate activity. In contrast, moxifloxacin and rifampicin had a low activity, while streptomycin, spiramycin and cephalosporines were ineffective. As a result, we come to the conclusion that a combination between one effective quinolone and doxycycline has a good efficacy against Brucella. Further in vivo studies are necessary to support this suggestion.

Detection of quinolone-resistance mutations in Salmonella spp. strains of epidemic and poultry origin

Mutations into codons Aspartate-87 (62 percent) and Serine-83 (38 percent) in QRDR of gyrA were identified in 105 Salmonella strains resistant to nalidixic acid (94 epidemic and 11 of poultry origin). The results show a high incidence of mutations associated to quinolone resistance but suggest association with others mechanisms of resistance.

Residuos de tetraciclina y quinolonas en peces silvestres en una zona costera donde se desarrolla la acuicultura del salmón en Chile / Residues of tetracycline and quinolones in wild fish living around a salmon aquaculture center in Chile

La presencia de antibacterianos y antiparasitarios residuales fue investigada en muestras de carne de peces silvestres de consumo humano pescados alrededor de un recinto de acuicultura en Cochamó (41° 29' S; 72° 19'W), X Región, Chile. Esta investigación demostró que peces silvestres, incluyendo róbalo (Elginops maclovinus), cabrilla (Sebastes capensis) y truchas de vida libre (Oncorhynchus mykiss), ingieren alimento artificial para salmón y que la carne de algunos ejemplares de estos peces contienen tetracicilina y quinolona en cantidades detectables. Estos resultados sugieren que el uso de antibacterianos en la acuicultura del salmón, como ha sido demostrado en otros países, tiene efectos ambientales que se proyectan más allá de los recintos de acuicultura. Se indica que dada la relevancia de estos hallazgos para la salud humana y animal, el ambiente requerirá de estudios más amplios y detallados para implementar futuras regulaciones del uso de antibacterianos en acuicultura.

The presence of residues of tetracycline, quinolones and antiparasitic drugs was investigated in wild fish captured around salmon aquaculture pens in Cochamó, Region X, Chile. Residues of both antibiotics were found in the meta of two species of wild fish that are consumed by humans, robalo (Elginops maclovinus) and cabrilla (Sebastes capensis) . These findings suggest that the antibiotic usage in salmon aquaculture in Chile has environmental implications that may affect human and animal health. More studies are needed in Chile to determine the relevance of these findings for human and animal health and the environment to regulate this use of antibiotics.