Ciencias Agropecuarias
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Item Presencia de genes de resistencia a los antibióticos en Escherichia coli aislada de carne de pollo que se expende en el cantón de Ambato(2024-11) Gómez Aillòn Diego José; Cruz Quintana Sandra MargaritaAntimicrobial resistance is currently a public health problem in Ecuador. It seems that the indiscriminate use of antibiotics in animal production is a way for the development of this resistance. E. coli is a pathogen that affects animals and people and has developed resistance to several antibiotics in recent years. Cephalosporins and quinolones are used in broiler production and are two families of antibiotics to which strains of E. coli They have shown resistance. This resistance is mainly mediated by resistance genes such as gyrA and blaCTXM. Studies have searched for these mechanisms in E. coli strains from chicken meat, as it is one of the main meats consumed in Ecuador. The present study aimed to evaluate the antibioresistance and the presence of resistance genes of 31 Escherichia coli strains to antibiotics. Antibioresistance was carried out by the agar diffusion method and the search for resistance genes through PCR. In this way, a Kruskall Wallis test and the Mann Whitney U test were performed for the antibioresistance data, and for the genes, prevalence and a chi-square test were analyzed, with a 95% confidence interval. The results for the antibioresistance of the strains were resistant to ciprofloxacin 45.1% and ceftriaxone 25.8% and the antibiotics that provided greater susceptibility were gentamicin and amikacin with a p≤0.05. For the gyrA genes of CIP and blaCTXM of CX there were prevalence of 100% and 96.77% respectively. When comparing the difference between the origin of the sales center and the resistance gene, there was no statistical difference p≥ 0.05 in both genes. Finding resistance to antibiotics gives us a warning that the indiscriminate use of these in the country's animal production could be an important factor for this resistance. The prevalence of genes is a guide to the main resistance mechanism that could be used by E. coli, such as mutations in genes and genes that encode enzymes that provide resistance. And finally, finding antibioresistance and resistance genes in chicken meat in the Ambato canton is a warning of the possible risk of transmissibility of resistant strains that could generate resistance to antibiotics in people. However, the current data in Ecuador on this possibility is not yet being studied with the importance of the case. Therefore, studies like this open a gap for us to analyze everything that antibioresistance implies and the repercussions on public health in Ecuador.Item Determinación de los agentes predominantes bacterianos presentes en la enfermedad periodontal moderada y severa en Canis lupus familiaris alimentados con dieta mixta(2023-09) Vivanco Cuenca, Daniela Liseth; Lozada Ortiz, Jenny PiedadThe objective of this research was to determine the predominant bacterial agents present in moderate and severe periodontal disease (PD) in Canis lupus familiaris older than 4 years old fed a mixed diet. The study was carried out in the Veterinary Medical Center of the Pueblo Unido neighborhood in the city of Quito with the collection of dental plaque samples from 33 canines diagnosed with moderate and severe PD. The samples were collected with a Gracey Curette and cytological brush and were immediately transported in a red cap tube without additive to the laboratory. Colony forming unit (CFU) counts were performed on plate count agar (PCA), the two bacteria with the highest number of CFU/gr were isolated on culture media and biochemical tests were performed for identification. In canines diagnosed with moderate PD, bacteria such as Staphylococcus aureus (68.4%), Escherichia coli (68.4%), Proteus mirabilis (42.1%), Proteus vulgaris (10.5%) and Streptococcus salivarius (10.5%) were isolated. Similarly, in those with severe PD, species such as S. aureus (92.9%), E. coli (71.4%). P. mirabilis (14.3%), P. vulgaris (14.3%), and Enterococcus faecalis (7.1%). Finally, it was evaluated whether the presence of these bacteria is dependent on the degree of moderate or severe progression of PD and the relationship with the CFU/gr count. It was concluded that there was no statistically significant difference (p=0.975) between the isolated bacteria and the degree of PD, however, there was a statistically significant difference (p<0.0001) in the presence of bacteria in relation to the means of CFU/gr count.Item Determinación de antibiorresistencia de cepas de Escherichia coli aisladas en carne de pollo en la ciudad de Ambato, provincia de Tungurahua(2023-09) Castro Velasco, Katerin Anahí; Cruz Quintana, Sandra MargaritaEscherichia coli presents significantly high and well-developed resistance mechanisms towards antibiotics, representing a worldwide problem for public health, one of its transmission routes is in food of animal origin, noting that the dissemination is facilitated by poor hygiene and Sanitary control in slaughterhouses. The objective of the present investigation was to determine the antibioresistance of Escherichia coli strains isolated in chicken meat in the city of Ambato, Tungurahua Province. We worked with a total of 31 E.coli strains subjected to two research phases, the first to establish the Resistance, Intermediate or Sensitivity of the strains to the group of antibiotics: Amikacin, Gentamicin, Amoxicillin + clavulanic acid, Ceftriaxone, Ciprofloxacin through of the Kirby Bauer phenotypic method (Disc-plaque), the results were subjected to the Kruskal Wallis test complemented with the Mann Whitney U test, obtaining a CIP Resistance: 45%, CRO: 25.8%, AMC: 9.8%. , CN and AK: 0%, following CLSI 2023 cut-off points. In the second phase, the MIC (Minimum Inhibitory Concentration) of the resulting strains sensitive to the group of antibiotics in phase 1 was determined, the results were analyzed descriptively through of curves establishing the MIC of AK: 18-20 μg/ml CN: 18-22 μg/ml CIP: 19-21 μg/ml CRO:18- 22 μg/ml AMC: 28-33 μg/ml, following points of CLSI 2023 cutoff. The main resistance mechanisms of Escherichia coli to beta-lactams are beta-lactamases, to aminoglycosides mutations in amino and hydroxyl groups and alteration of target sites and in fluoroquinolones chromosomal mutations in DNA gyrase and topoisomerase occur, all of these methods developed by E.coli block the action of drugs. In conclusion, it was determined that E. coli shows a marked resistance to antibiotics thanks to resistance methods that the bacteria presents through enzymes and mutations.Item Calidad microbiológica de canales de pollo (Gallus gallus) en centros de faenamiento que suministran al cantón Ambato(2023-03) Manzanillas Miranda, Carolina Beatriz; Cruz Quintana, Sandra MargaritaFoodborne diseases (ETA) are a public health problem, in which bacteria have a higher incidence in diarrheal infections. Consequently, food safety is a significant problem worldwide, because, in the areas where chicken carcasses are obtained, there may be different points of cross contamination, being Staphylococcus aureus, Salmonella spp, Vibrio cholerae, Escherichia coli, Clostridium botulinum, Campylobater spp and Listeria monocytogenes the most frequent pathogens. Therefore, the objective of this work was to evaluate the microbiological quality of chicken carcasses (Gallus gallus) in slaughterhouses that supply the Ambato canton. The methodology that was used consisted of taking 45 samples of chicken meat, taking 5 samples per slaughterhouse; with which we worked in the Microbiology Laboratory of the Technical University of Ambato, counting mesophilic aerobics, sowing on selective agar such as Modified Chromogenic Listeria agar, MacConkey agar, agar, Buffered Peptone agar, Nutrient agar, Xylose Lysine Deoxycholate agar, Triple Sugar Iron agar, Gram stain, oxidase and catalase test. The result that was obtained in the 45 samples collected in the nine different slaughterhouses were, 11 samples with values of 1.05 x 106 CFU/g, 4 with values of 9.18 x 106 CFU/g and 30 samples with values greater than 1.0 x 107 CFU/g, with an average count of 4.03 x 107 CFU/g. In the same way, of the 45 samples were identified, 2 samples of Listeria spp, 16 samples of Salmonella spp and 42 samples of Escherichia coli. On the other hand, the risk factors with the highest incidence in the contamination of chicken meat were the microbiological quality of drinking water, operator health safety, staff training, the scalding phase, evisceration, pre-cooling, cooling of the chicken meat and its safety.Item Detección de Salmonella spp y Escherichia coli en muestras de carne de pollo que se expende en el cantón Ambato(2023-03) Gómez Usiña, Jennifer Patricia; Cruz Quintana, SandraThe greatest health risk associated with the consumption of chicken meat are foodborne diseases, due to its ability to serve as a vehicle for pathogenic bacteria, it is important to study it. The present research aimed to detect the presence of Salmonella spp. and Escherichia coli in samples of chicken meat, which is sold in Ambato. The methodology that was used consisted of taking 90 samples of chicken meat, 45 samples from informal stores and 45 samples from authorized stores; with which we worked in the Microbiology Laboratory of “Universidad Técnica de Ambato”, counting aerobic mesophiles, sowing in selective agars such as MacConkey and XLD, Gram staining and biochemical tests such as TSI, oxidase and catalase. The result obtained was that all the samples taken from the stores were contaminated with some type of bacteria. Contaminated with Escherichia coli 8.1% of certified retail stores and 84.4% of informal stores; followed by Salmonella spp. with it 8.1% of certified premises and 48.8% of informal premises. The result of the count of aerobic mesophiles in the chicken meat samples demonstrates the relationship that exists between the minimum conditions of a retail stores and the microbiological quality of the chicken meat, highlighting that the hygiene of the premises, meat conservation and measures protection of the seller in the informal outlets have a significant difference with respect to the certified stores.Item Correlación entre los niveles de pH y la proliferación de Escherichia coli en las canales bovinas(2022-02) Espín Montesdeoca, Ricardo Armando; Guevara Freire, DeysiThis study examined the correlation between pH levels and the proliferation of Escherichia coli in bovine carcasses. A total of 23 bovine carcass samples were analyzed. The animals were monitored from the moment they entered the cattle resting pens at the Ambato Municipal Camal. In addition to the identification of the animals, the management indicators (direct and indirect), the sensitivity indicators, and the cleaning indicators were recorded. One hour after slaughter, the pH was measured with the help of a HANNA HI 99163 pH meter for meat products, using the ISO 2917 rule, at three points comprised by the Longissimus dorsi muscle located between the 8th rib and 2nd lumbar vertebra. To collect E. coli samples, the Whirl-pack method comprising sites S1, S2, and S3 was used. Traces of E. coli were then processed in the slaughterhouse's laboratory. Results were obtained that 91.3% of cattle were prodded with an electric prod, 39.1% vocalized, 21.7% slipped, and 4.3% fell, in the cattle handling indicators. The percentage of that category that fell to the first shot and the percentage that was hit by the guillotine door was 78.3% and 43.5%, respectively. The time between the shot and bleeding was approximately 88 seconds. According to the results, 30.4% of the participants were sensitive to vocalization, 21.7% showed rhythmic respiration, 17.4% had corneal reflexes, and 13% elevated their heads. Cleaning indicators showed that 17.4% of workers had washed their hands. The percentage of bruises on category one corpses was 26.1%, while the percentage of bruises on category two corpses was 8.7%. During the cooling process, the corpses remained in the cooling room for 60.2 minutes. The pH levels found in the bovine carcasses averaged 6.73 ± 0.09, 39.2% of the pH values were in the range of 6.64 - 6.82, 30.4% in the range of 6.45 to 6.63, and the remaining 30.4% in the range from 6.83 to 7.01. The proliferation of E. coli occurred in 78.3% of the processed samples, with a mean of 0.23 ± 0.08 log CFU / cm2. 60.9% of the positive samples were found in the range of 0.02 to 0.82 log CFU / cm2, 17.4% were in the range -0.80 - 0.01 log CFU / cm2 and 21.7% presented its absence. Pearson's correlation was r = 0.84 very strong and elevated between pH levels and proliferation of E. coli.Item Perfiles fenotípicos de resistencia a antibióticos en aislados de Escherichia coli de origen canino de la ciudad de Ambato(2021-03) Robalino Proaño, David Guillermo; Calero Cáceres, William RicardoCurrently, antibiotic resistance crisis is a global concern, both in human and veterinary medicine. The uncontrolled evolution of the resistance mechanisms in bacteria has had a negative influence on effective therapeutic options. This research has focused on the enterobacterium Escherichia coli, focused on the detection of the extended espectrum beta-lactamases (ESBL) and AmpC type beta-lactamases, which represent variants with resistance mechanisms of great clinical impact in Gram negative bacteria. In Ecuador, few epidemiological data are available on E. coli producing ESBL and AmpC in animals. In the present study, the phenotypic profiles of resistance to antibiotics were evaluated in a collection of isolates of E. coli of canine origin from the city of Ambato, using the Kirby & Bauer disk diffusion method, detecting that 72.2% (57/79) of isolates have an ESBL phenotype and 81% (64/79) of isolates with manifestation of the AmpC phenotype. The percentages of resistance for the evaluated antibiotics was: Ampicillin 100% (79/79), Vancomycin 100% (79/79), Cefotaxime 98.7% (78/79), Erythromycin 98.7% (78/79), Ceftriaxone 97.5% (77 / 79); and phenotypic sensitivity patterns for antimicrobials such as: Meropenem 98.7% (78/79), Imipenem 96.2% (76/79) Piperacillin / Tazobactam 84.8% (67/79), Cefoxitin 83.5% (66/79), and Amikacin 82.3% (65/79). The patterns of bacterial resistance detected and the differences between the origin of the isolates will allow to use this background information for retrospective studies of antimicrobial resistance.Item Determinación de la presencia de genes de resistencia a betalactámicos y evaluación de diversidad clonal en aislados de Escherichia coli de origen canino de la ciudad de Ambato(2020-03-19) Garcés Arias, Xiomara Sthefanny; Calero Cáceres, William RicardoThe evolution and spread of multiresistant bacteria is considered prioritary, considering their impact in public health and animal safety worldwide. Therefore, the detection of reservoirs, routes of dissemination and evolution of these bacteria is aimed in order to limit their negative effects on public and animal health. Extended-spectrum-betalactamase Escherichia coli producer is considered an emerging health problem. In Ecuador, there are few available data on the molecular epidemiology of those of ESBL- E. coli producing bacteria in animals and environment. In the present study, the presence of beta-lactam resistance genes was evaluated as well as the clonal diversity of E. coli isolates of canine origin from the city of Ambato. The highest presence of beta-lactam resistance genes occurred in dogs with owners, in contrast to stray dogs. The most frequently detected genes were blaCTX-M and blaTEM. The blaSHV and blaCMY genes had a lower detection rate, however, these were found in combination with the blaCTX-M and blaTEM genes, indicating that they are isolates with considerable virulence. The analysis of clonal relatedness through the amplification of palindromic repetitive regions (GTG5) detected that the isolates comes from different lineages, discarding the hypothesis that suggest the presence of multi-resistant clones with a common ancestor, suggesting that their genotypes were acquired through recent horizontal genetic transmission. Dogs can act as a silent reservoir of microorganisms with antibiotic multiresistance genotypes. It is necessary an extended review of waste management procedures is necessary, as well as interactions with humans and animals under certain circumstances like immunological disorders.Item Efecto antimicrobiano in vitro del aceite esencial de eucalipto (Eucalyptus spp.) sobre cepas certificadas de Escherichia coli Y Staphylococcus aureus(2018) Morocho Núñez, María José; Montero, MayraThe objective of the research project was to evaluate in vitro the antimicrobial effect of Eucalyptus essential oil (Eucalyptus spp.) on certified strains of Escherichia coli ATCC25922 and Staphylococcus aureus subsp. aureus ATCC292113; in which dilutions of eucalyptus oil were evaluated 30%, 60% and 90% with 98% ethanol. The Minimum Inhibitory Concentration was determined using the broth microdilution method, each bacterial inoculum was standardized to 0.5 of the MacFarland scale in the spectrophotometer, obtaining as a result that the 60% and 90% eucalyptus oil tubes did not present turbidity for Escherichia coli, similar characteristic that presented the strain Staphylococcus aureus subsp. aureus in the 60% and 90% dilutions, the Bactericidal Minimum Concentration was also determined by seeding the dilutions mentioned above in the Mueller-Hinton agar in which no growth of colonies was observed at 60% and 90% for the two strains evaluated. The disc diffusion test showed the sensitivity of the evaluated strains to eucalyptus oil, presenting for Escherichia coli a minimum halo of 10.25mm at 30% and a maximum diameter of 10.95mm at 90% while for the Staphylococcus aureus strain subsp. aureus obtained a minimum value of 10.95mm at 30% and a maximum value of 14.45mm at 60% dilution. Eucalyptus essential oil (Eucalyptus spp.) Has antibacterial properties for Gramnegative Escherichia coli and Gram-positive bacteria Staphylococcus aureus sbsp.aureus.Item Efecto antimicrobiano in vitro del aceite esencial de orégano (Origanum vulgare) sobre cepas certificadas de Escherichia coli Y Staphylococcus aureus(2018) Lopez Rivera, Edwin Andrés; Montero, MayraThe objective of the research project was to evaluate in vitro the antimicrobial effect of oil of oregano (Origanum vulgare) on strains of Escherichia coli and Staphylococcus aureus subsp. aureus, in which concentrations of 30%, 60% and 90% were evaluated in dilution of oregano plus tween 80 oil. The Minimum Inhibitory Concentration was determined for the Escherichia coli strain, where turbidity was present at 30% in all its established hours, while at 60% and 90% it did not present turbidity, on the other hand, for the strain Staphylococcus aureus, no turbidity was present in any of its concentrations; The Bactericidal Minimum Concentration was also determined on the strain Escherichia coli and Staphylococcus aureus, where the 30% Escherichia coli strain presented colony formation at all established times, while at 60% and 90% there was no bacterial growth; On the other hand, for the strain Staphylococcus aureus, there was no bacterial growth in any of the three concentrations. Regarding the sensitivity halos in the Escherichia coli strain, a minimum diameter of 13.01mm at 30% and a maximum value of 17.62mm at 60% was obtained, determining that the oil of oregano prevents the growth of bacteria from its minimum value at 30%, on the other hand, in the Staphylococcus aureus strain, a minimum value of 13.25mm was obtained at a concentration of 30% and a maximum value of 25mm at 90%. In the analysis of variance the results obtained in the treatments at 60% and 90%, are not significantly different (p = <0.05), being statistically equal with values of 17.62 mm and 16.05 mm respectively, in relation to 30% with a value of 13.01 mm in halos of inhibition for the strain Escherichia coli, while the dilutions of the treatments at 30%, 60% and 90% against the strain Staphylococcus aureus are statistically different (p = <0.05) with values of 13.25 mm, 16.35 mm and 25.00mm of inhibition halos.