Browsing by Author "Nieto Ramírez, Luisa María (Directora)"
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Item Detección molecular de Leptospira spp en roedores plaga del Zoológico de Cali(Universidad Santiago de Cali, 2020) Jaramillo Ruiz, Felipe Antonio; Nieto Ramírez, Luisa María (Directora)Leptospirosis is a zoonosis caused by bacteria of the genus Leptospira spp, with rodents being its best known reservoir. The Zoo of Cali controls rodent plagues which potentially keep the circulation and presence of this bacterium active. To prove the presence of Leptospira spp and its pathogenic species among the plague rodents of the Cali Zoo, a total of 32 rats were captured between May and August 2019. Using samples of renal tissue, it was carried out the amplification by PCR of rrs and lipL32 genes, detecting a positivity of 12.5% for both genes. lipL32 also constitutes a molecular marker of pathogenicity whose product was also analyzed by Sanger sequencing, identifying L. interrogans species with a similarity of 100%. The results achieved allow confirming the presence of pathogenic species Leptospira spp in the renal tissue of rodents that circulate in the area of Cali Zoo, generating an evidence of the potential risk of transmission to other animal species, including visitors or workers of the institution.Item Moluscos de el Charco Nariño: Una mirada desde la exploración bioquímica(Universidad Santiago de Cali, 2020) Caicedo Perlaza, Yaneth Angélica; Nieto Ramírez, Luisa María (Directora); Pantoja Pulido, Kriss Dayana (Directora)Mollusks are non-segmented soft-bodied invertebrates protected by a shell and constitute the basis of the diet of many communities. Our objective was to determine the biochemical composition of two species of mollusks Littorina zebra (piacuil) and Anadara tuberculosa (piangua), which are an important nutritional source in El Charco, Nariño. In total, 30 samples of L. zebra with average sizes of approximately 3cm were collected, and 30 samples of A. tuberculosa differentiated by sex, identifying 13 females with average lengths of 8.4cm and 17 males with average lengths of 6.3cm. Mud samples were also collected from the mollusk habitat, from which the percentage of humidity and pH was determined. The bodies of the mollusks were separated from their shells and then lyophilized to carry out the determination of proteins by the method of BCA (Bicinconinic Acid) and polyacrylamide electrophoresis, extraction of total lipids by the Folch method and evaluation of these by layer chromatography fine (TLC). Finally, the total DNA extraction was carried out using a commercial kit and the quantification and evaluation of its quality using the Nanodrop equipment. In the soil samples a humidity of 23.4% and a pH of 6.25 were observed on average. An average value of total proteins of 3.27 g (± 0.65) was obtained for male A. tuberculosa, 3.73 g (± 0.14) for female A. tuberculosa and 4.69 g (± 0.11 ) for L. zebra per 100 g of lyophilized tissue, values that are lower than those reported in the literature. The amount of total lipids extracted was determined, obtaining an average of 2.1 g (± 0.8) for L. zebra, 2.45 g (± 0.3) for female A. tuberculosa and 2.35 g (± 0.7) for Male A. tuberculosa per 100 g of lyophilized tissue, values very similar to those found in the literature. A qualitative evaluation of lipids was carried out by TLC (Thin Layer Chromatography), where the presence of colored bands that demonstrated the oxidation of organic compounds and the presence of active compounds was observed. Finally, the DNA for L. zebra and A. tuberculosa male was successfully extracted, however, for the female it was not possible to obtain a DNA solution of satisfactory quality, for which the use of another methodology is recommended to eliminate pigments that affect Nanodrop quantification. This work was able to corroborate the ancestral knowledge of the Pacific communities that this type of species has a high protein content, particularly the L. zebra species and nutritional value; Therefore, it will be possible to bring documented information to the communities that allows them to validate their ancestral beliefs and to make good use of these natural resources.Item Validación secundaria del método de filtración por membrana para la detección de microorganismos indicadores de contaminación en muestras de agua de uso farmacéutico(Universidad Santiago de Cali, 2019) Guarnizo Mejía, Mariana; Nieto Ramírez, Luisa María (Directora)Purified water is supplied in the manufacture of non-parenteral products in the pharmaceutical industry. This input requires a microbiological analysis due to the susceptibility of microbial contamination in the distribution process, thanks to the presence of biofilms, among other factors. The filtration by membrane allows detecting the causes of said contamination and for this reason it is important to verify this methodology in the industry so that it can be used as a method of quality control in the plant located in the urban area of Cali. The present work was carried out in the pharmaceutical company Sanofi Cali, where the challenge organisms used in each stage of the process were evaluated, respectively the strains ATCC (American Type Culture Collection) Escherichia coli ATCC 8739, Staphylococcus aureus ATCC 6538 and Pseudomonas aeruginosa ATCC 9027. The stages of the process were a) inoculum standardization, b) productivity and selectivity tests of the culture media (Plate Count Agar, Chromocult and Cetrimide), c) development of the secondary validation technique of the membrane filtration method. The comparison of the results obtained by two analysts was included during the validation process. The effectiveness of validation for S. aureus and E. coli and non-effectiveness in P. aeruginosa was demonstrated.