Microbiología

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Estudio de la micota ambiental de tres edificios de la universidad Santiago de Cali, Colombia
(Universidad Santiago de Cali, 2019) Lasso Ceballos, Ana María; Caicedo Bejarano, Luz Dary
The quantification of the environmental load in the indoors of the buildings (1, 3 and 4) of the University Santiago De Cali, Colombia and in their corresponding responses to the results to obtain a load value. A volumetric method and Dicloran Bengal chloramphenicol (DRBC) and CHROMagar Cándida culture media were used for sampling. The identification of the fungi was carried out based on macro and microscopic characteristics. Parallel to the sampling the relative humidity and temperature of each space studied was determined. This study was conducted for quarterly periods during twelve months (March 2018 to march 2019). In the study of environments of the three buildings was detected a total of 82393 colony forming units (CFU/m3) of which 60053 CFU/m3correspond to indoor environment. The results obtained showed that it was not possible to establish a correlation between the temperature and the CFU/m3 found inside the three buildings, but for the relative humidity in relation to building 4.The genera Cladosporium, Fusarium, Penicillium and Aspergillus were identified as the most prevalent allergenic fungi in this study. The results in the quantification revealed that the fungal burden exceeds the limits accepted for internal environments (500 CFU/m3) according to the World Health Organization (WHO).
Biota fúngica en fosas nasales de trabajadores de tres edificios de la universidad Santiago de Cali y su relación con síntomas de alergias respiratorias
(Universidad Santiago de Cali, 2019) Castillo Tellez, Beatriz Angélica; Caicedo Bejarano, Luz Dary
Allergenic fungi enter through the air to nostrils and may be responsible for some respiratory allergies. The objective of the present study was to determine the relationship between allergies diagnosed and undiagnosed according to the results obtained from the adapted survey of ISAAC (International Study of Asthma and Allergies in Childhood) and the fungal load of the nostrils of workers of three buildings of the Santiago de Cali University. The samples taken from the nostrils were made with sterile swabs and the subsequent sowing was done on Sabouraud dextrose agar, sunflower seed agar and Candida CHROMagar. In 100% of the workers (126) who participated in the study, allergenic fungi were isolated. The highest prevalence of allergenic fungi in the workers of the different buildings was obtained by the workers of block 1, finding significant statistical differences, for the genus Fusarium sp, (p=0,0018 Kruskal Wallis) and Penicillum sp, (p=0,0080 Kruskal Wallis), but not for Cladosporium sp, Alternaria sp and Aspergillus sp; likewise, the biggest fungal load was presented by the workers of this building. When analyzing the fungal 6load found in the nasal passages and its relation with the presence of symptoms of respiratory allergies, it was established that there was significant statistical association with the genus Fusarium sp (p=0,0430 Wilcoxon Mann Whitney) and not for Cladosporium sp (p=0,1918, Wilcoxon Mann Whitney) which was the most frequently isolated genus.
Caracterización molecular de aislados de Enterobacter cloacae resistentes a carbapenemes portadores de los genes blaKPC, blaNDM, blaVIM y blaOXA-48, provenientes de la red de laboratorios públicos y privados del Valle del Cauca
(Universidad Santiago de Cali, 2019) Guerrero De La Cruz, Daniela Andrea; Restrepo Falco, Aura
The production of beta-lactamase enzymes by some species of enterobacteria is a serious public health problem because it causes the loss of therapeutic efficacy to beta-lactam antibiotics, which are currently used as the first alternative for the management of infections caused by Gram-negative bacteria. In this work, the molecular characterization of 32 phenotypically resistant Enterobacter cloacae clinical isolates of cephalosporins and carbapenemes, from a private and a public entity, located in the city of Cali, was carried out. In order to detect the genes involved in resistance to these antibiotics, the polymerase chain reaction technique was used to amplify the genes encoding carbapenemases: KPC, NDM, VIM and OXA-48. It was obtained that 50% (16/32) of the isolates are carriers of the blaKPC gene, while the presence of blaNDM, blaVIM and blaOXA-48 genes was not detected. The results of this study indicate that the allele that circulates in the isolates is KPC-2 and that they are not genetically related. The results indicate that 81% of patients infected with the enterobacteria carrying KPC carbapenemase come from private medical centers, 81% are male and 44% of those affected are older adults (60 years and older). Knowing the genes involved in carbapenemes resistance in these clinical isolates is an important step towards the development of specific strategies to prevent the spread of this multi-drug resistant pathogen in the municipality of Cali.
Caracterización fisicoquímica, sensorial y micológica del kéfir de agua en jugos de frutas tropicales
(Universidad Santiago de Cali, 2019) Pedraza Morales, Yiham Stephanie; Ramírez Navas, Juan Sebastián
Kefir contains beneficial properties that promote health, which is why microorganisms isolated from kefir are studied for the development of functional foods. In the present study, the physicochemical, sensory and growth characteristics and phenotypic characterization of fungi present in water kefir in tropical fruit juices were determined. This experiment was done under a factorial design corresponding to the combination of two factors with three blocks and three replicas: type of substrate (pear juice, passion fruit and water with panela) and sucrose concentration (25%, 50% and 100%). The response variables were: acidity, pH, Brix degrees. For the microbiological analyzes, decimal dilutions were made, they were counted on the second and fifth day of incubation. The identification was made with API 20C and Vitek 2 (Biomérieux). Three yeast strains found from higher to lower number of CFU/mL Saccharomyces cerevisiae 1, Saccharomyces cerevisiae 2 and Zygosaccharomyces spp were finally isolated with the sensory analysis it was determined that the passion fruit drink was the one that had the best acceptance and preference on the part of consumers.
Evaluación del efecto de la plata coloidal, en la remoción de escherichia coli y la obtención de un agua salubre a partir de filtros caseros
(Universidad Santiago de Cali, 2019) Carabalí Ramos, Juan José; Pérez Vidal, Andrea; Rivera Sánchez, Sandra Patricia; Pérez Vidal, Andrea
Water is a natural and indispensable resource for life, for this reason it is very important to supply it, in a drinkable, affordable and good quality way to be supplied to the community in general, fulfilling the physicochemical characteristics, Microbiological and sanitary concept, established in the standard, Resolution 2115 of 2007. However, despite drinking water being a right, many communities, especially rural ones, still do not have access to quality water; presenting Diarrheal Watered Diseases (EDA) and Foodborne Diseases (ETA) which are induced by the intake of unhealthy water, among other factors that help to spread the disease. For this reason, alternative techniques and / or processes for water treatment have been sought, among which techniques such as home filtration systems for obtaining drinking water have been highlighted. These filters are composed of various materials, such as colloidal silver, activated carbon, which possess antimicrobial potential and / or retention capacity of substances and microorganisms. In order to mitigate the EDA and ETA, the REPLACOL microenterprise developed and provided the EKOFIL filtration systems impregnated with colloidal silver, and was sought as: Objective: To evaluate the effect of colloidal silver on the removal of Escherichia coli in obtaining a safe water from homemade filters. Methodology: The process of water purification was carried out in the facilities of the Santiago de Cali University, subjecting the home filters to certain concentration of synthetic water, that is; 103, in accordance with the inoculation provided from E. coli strain ATCC 25922 employed by membrane filtration (FM) for the diagnosis of water quality at certain times of 1, 8 and 24 hours of silver contact with filtered effluent and residual silver. Through statistical analysis (Kruskal-Wallis and post anova) they were obtained as: Results: The determination of the best contact time in conjunction with the best filtration rate by home filters therefore determined the filter defined as E 1.0 which managed to remove up to 3.9 logarithmic units from the initial synthetic water. Conclusion: The efficiency of removal of the E 1.0 filter with respect to the other systems is observed, determining that the best contact time occurs at a time of 8 hours with a filtration rate of 1L / h where the removal reaches up to 100 % of effectiveness
Caracterización de bacterias asociadas a la rizosfera de plantas de yuca (manihot esculenta crantz) sometidas a estrés por déficit hídrico
(Universidad Santiago de Cali, 2019) Galindo Espinal, Diana Marcela; Zapata Gualtero, Tatiana
Cassava (Manihot esculenta Crantz) is one of the most important energy sources in the tropical and subtropical regions of the world. During the last years, its production has resulted in extreme environmental conditions. Plant growth promoting rhizobacteria (PGPR) are a type of bacteria that colonize the roots and a symbolic relationship with the plant, favoring their growth and resistance even in conditions of abiotic stress. In the present study, 124 bacterial components of cassava plants M. esculenta are characterized, of which 26 rhizobacteria are identified in plants over a period of 20 days. The analyzes of the abundance of the most predominant genera are: Bacillus, Achromobacter, Pseudomonas, Sphingomonas and Streptomyces. These results are important for bacterial communities such as cassava cultivation and its relationship with the response of plants to abiotic stress conditions.
Comparación de dos técnicas para la captación de esporas fúngicas ambientales en un campus universitario
(Universidad Santiago de Cali, 2019) Delgado Ñañez, Cristhian; Caicedo Bejarano, Luz Dary
The concordance of two fungal spore uptake techniques in the environments of three buildings of the Santiago de Cali University campus was evaluated. The gravimetric method of spore uptake was compared with the Omeliansky formula and the volumetric method (Air Ideal 3P). The environmental sampling was carried out quarterly for a period of 12 months using Dicloran Rose Bengal Chloramphenicol agar (DRBC). 64497 CFU / m3 were accounted for, of which 34930 CFU / m3 were isolated with the volumetric method and 29567 CFU / m3 with the gravimetric method. No significant statistical differences were observed with the two techniques used (p = 0.0739), with a significance level α = 0.05. The predominant genera obtained with both methods were Cladosporium, Penicillium, Aspergillus and Fusarium, which suggests that although the gravimetric method is not as efficient as the volumetric method, it is reliable for estimating the degree of environmental fungal contamination in a similar way to the volumetric method.
Evaluación de la remoción de salmonella spp. a partir de dos calidades de aguas sintéticas tratadas con filtros caseros
(Universidad Santiago de Cali, 2019) Castaño Hincapié, Ana Valentina; Rivera Sánchez, Sandra Patricia
Water is the main resource that abounds in the Earth. It is not renewable and has great importance, both for human consumption and for the rest of living beings. It is used for domestic uses or other purposes. The low coverage of aqueduct and sewerage in rural areas of Colombia, added to the fact that the implementation of potabilization in these areas entails a high cost, have favored the creation of new technologies that imply minor resources and accessibility, such as the homemade filters.
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
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.
Susceptibilidad a los antimicóticos de levaduras aisladas de sistemas de aguas de la ciudad de Santiago de Cali, Colombia
(Universidad Santiago de Cali, 2019) Osorio Vanegas, Lizeth Stefania; Caicedo Bejarano, Luz Dary
Water systems become a high capacity of microorganisms. The objective of this work was to evaluate the susceptibility to the antifungals of yeasts present in the wastewater and natural waters of the city of Cali. Samples were collected from two types of water: potable (Río Melendez, Puerto Mallarino) and residual (Canal Sur, Planta de Tratamiento Cañaveralejo-PTAR). Yeasts were isolated in sabouraud and CHROMagar Candida in the commercial identification with API 20 C AUX (BioMérieux) and in the sequence analysis of the ITS1- 5.8S-ITS2 region. The minimum inhibitory concentration (MIC) of fluconazole and amphotericin was determined by the microdilution method, and the influence of some physicochemical parameters and heavy metals was correlated with the possibility of yeasts and antifungals. With the phenotypic tests, the 15 yeast species are identified and the molecular biology techniques six species, the morphotype of the property that can belong to the new species of the genus Candida. 100% of the strains were sensitive to Amphotericin B, and 44.3% were sensitive, 19.23% were sensitive dose-dependent (DDS) and the remaining 25% were resistant to fluconazole. Finally, a correlation has been found between the minimum inhibitory concentration, some physicochemical parameters and the concentration of heavy metals on the antifungal sensitivity of yeasts associated with aquatic systems in the city of Cali.
Evaluación de la actividad antimicrobiana de péptidos catiónicos vehiculizados en nano liposomas funcionalizados, frente a bacterias patógenas de alimentos.
(Universidad Santiago de Cali, 2019) Cantor Pareja, Stefania; Oñate, Jose Fernando
During recent years the resistance of pathogens present in food to antibiotics has been demonstrated. That is why, in this research project, the antimicrobial activity of the synthetic peptides Alyteserin 1C (WT) and its mutant product (ΔM) was evaluated in bacterial strains such as Staphylococcus aureus ATCC25923, Listeria monocytogenes ATCCbaa751, Salmonella typhi ATCC25922. Use of two procedures: modification of the antimicrobial peptides and vehiculization of the peptides in liposomes coated with cationic polymer Eudragit E100. The results showed the antibacterial activity of the peptides against the bacteria. However, after the encapsulation of the peptides in liposomes coated with Eudragit E-100, the antibacterial activity was reduced from 1000 to 2000 times against the strains of Listeria monocytogenes ATCCbaa751 and Escherichia coli ATCC25922
Evaluación de la remoción de Escherichia Coli presente en dos calidades de aguas sintéticas tratadas con filtros caseros
(Universidad Santiago de Cali, 2019) Dávila Estupiñán, Andreína; Rivera Sánchez, Sandra Patricia
Introduction: Water is essential for life and all people must have a satisfactory, sufficient, safe and accessible supply; Likewise, it must comply with the microbiological and organoleptic characteristics established in the regulations, including the total absence of Escherichia coli, in order to be suitable for human consumption; However, a large percentage of especially rural areas do not have good quality water and have diseases for this reason, so resorting to new technologies such as household filters that help purify water, provides an alternative with tangible benefits for health. Objective: To evaluate the removal of Escherichia coli from two synthetic water qualities of EKOFIL and EKOFIL PLUS homemade filters. Methodology: The process for water potabilization was developed at the Santiago de Cali University and the Departmental Public Health Laboratory. Synthetic waters inoculated with the Escherichia coli strain ATCC 25922 at a concentration of 102 and 103 were used; and they were evaluated by the membrane filtration microbiological method. The proposed design was an experimental type study of a factor. For the evaluation of the removal of the homemade filters, an Anova was used. Results: For the two evaluated synthetic water qualities (102 and 103 CFU/mL) there were no significant statistical differences for the removal of Escherichia coli compared to filtration rates. However, the EKOFIL PLUS filters achieved a removal of the microorganism from 2.2 Log in the concentration of 102 CFU/mL and 3.0 Log for 103 CFU/mL, which can be attributed to the presence of the column with activated carbon especially with filtration rate of 2 h / L and 1 h / L respectively. Conclusions: Although the filters showed removal of Escherichia coli in the water after filtration, it fails to meet the criterion recommended by Colombian regulations, which is 0 CFU/100mL, regarding this microbiological indicator in some of the filtration systems and there was no clear trend.
Sensibilidad cutánea (skin prick test) a hongos alergénicos de los trabajadores de tres edificios de la universidad Santiago de Cali y su relación con los síntomas de alergias respiratorias
(Universidad Santiago de Cali, 2019) Calderón Quintero, Cristina; Caicedo Bejarano, Luz Dary
Se planteó un estudio epidemiológico observacional de corte transversal, se describe una relación a la exposición de hongos en ambientes intramurales y a la presencia de hongos alergénicos en fosas nasales. En el estudio participaron 126 trabajadores de la Universidad Santiago de Cali, quienes firmaron un consentimiento informado y contestaron la encuesta adaptada de ISAAC (The International Study of Asthma and Allergies in Childhood) para determinar la prevalencia de alergias diagnosticadas y no diagnosticadas. 71/126 (56,3%) trabajaban en el bloque 1; 24/126 (19,1%) en el bloque 3 y 31/126 (24,6%) en el bloque 4, La prueba “in vivo” Skin Prick Test (SPT) fue practicada a 75 trabajadores. Se encontró una prevalencia de SPT+ para Fusarium oxysporum de 10/75 (13,3%), Aspergillus fumigatus 5/75 (6,7%), Penicillium notatum 5/75 (6,7%), Alternaría alternata 5/75 (6,7%) y Cladosporium herbarum 1/75 (1,3%). Este estudio permitió establecer una posible asociación entre la presencia de hongos en los ambientes laborales y en las fosas nasales de los trabajadores con la respuesta positiva para la prueba SPT para Fusarium oxysporum
Comparación de diferentes medios de cultivo para el aislamiento de hongos alergénicos a partir de ambientes y fosas nasales
(Universidad Santiago de Cali, 2019) Concha Alarcón, Héctor Raúl; Caicedo Bejarano, Luz Dary; Chávez Vivas, Mónica
Different culture media were evaluated for the collection of environmental samples (Agar Dicloran Rosa de Bengala Cloranfenicol, CHROMagar Candida, Avena Agar and , Sabouraud Dextrosa Agar + 5% NaCl) and nasal fossa samples (Sunflower seed agar, SDA and CHROMagar Candida) of workers from three University buildings Santiago from cali. It was found that there were statistical differences, p>0.05, when CHROMagar Candida and Agar Dicloran Rosa de Bengala Cloranfenicol were compared with the volumetric method. , Sabouraud Dextrosa Agar + 5% NaCl recovered the lowest number of UFC / m3 , p <0.0001 when compared with the other media used with the gravimetric method and in the study of nostrils it was found that SDA was the culture medium where the higher counts P<0.0001. In this study, the Agar Dicloran Rosa de Bengala Cloranfenicol culture medium demonstrated that it is the medium of choice for the isolation of fungi from environmental samples and Sabouraud agar for clinical samples
Validación secundaria del metodo filtración por membrana para cuantificar la remoción de Escherichia Coli en muestras de aguas tratadas con filtros caseros
(Universidad Santiago de Cali, 2019) Martínez Rivera, Jorge Iván; Rivera Sánchez, Sandra Patricia
Con el fin de contribuir en la competencia para que laboratorio de la universidad Santiago de Cali pueda obtener certificaciones futuras en sus técnicas microbiológicas y con el apoyo de nuevas tecnologías de desinfección y tratamiento de agua proporcionadas por la micro empresa REPLACOL con sus dos modelos de filtros caseros de olla cerámica impregnado de plata coloidal (EKOFIL) y reforzado en su interior con una columna de carbón activado granular, también impregnada de plata coloidal (EKOFIL PLUS), diseñados para eliminar partículas suspendidas en el agua incluyendo microorganismos, se busco como Objetivo: realizar la validación secundaria de la técnica filtración por membrana para evaluar la remoción de Escherichia coli en muestras de aguas sintéticas provenientes de filtros caseros EKOFIL Y EKOFIL PLUS.
Evaluación de la capacidad inhibitoria de extractos de curuba y moléculas bioactivas sobre el crecimiento de mycobacterium smegmatis mc2155.
(Universidad Santiago de Cali, 2019) Anacona Sánchez, Paola Andrea; Aranaga Arias, Carlos Andrés
The intrinsic resistance that mycobacteria show to some disinfectants and antibiotics makes them a growing public health problem. Due to the fact that natural products and their derivatives have been shown to be a source of new antimicrobials, the effectiveness of hexanic, ethanolic and methanolic extracts of cascara and pulp of the fruit of the curuba (Passiflora mollisima) and some chalcone molecules synthesized by the Research Group of Heterocyclic Compounds of the Universidad del Valle (Colombia), on the growth of Mycobacterium smegmatis mc2155. The results showed that the hexanic extract of peel and a chalcone molecule, were able to inhibit the growing mycobacterium at concentrations of 1 mg/ml and 100 μM, respectively. This demonstrates the potential of curuba, to obtain molecules that, like the chalcones, serve as the basis for obtaining and designing new drugs.
Validación secundaria del número más probable para evaluar la remoción de salmonella spp. a partir de aguas sintéticas tratadas con filtros caseros
(Universidad Santiago de Cali, 2019) Lucero Huertas, Adriana Isabel
“Evaluación de la producción de carotenoides en levaduras nativas aisladas de la ciudad de Cali”
(Universidad Santiago de Cali, 2020) Villota Cobo, Sandra Viviana; Ramírez Castrillón, Mauricio; Martínez Garay, Carlos Andrés
In this study, the carotenoid production capacity in native yeasts isolated from the city of Cali was evaluated. 30 yeasts were selected, each of them was seeded in duplicate in MSS (solid pigment inducing medium), they were incubated for 5 days at room temperature (25°C). Pigments were extracted from the biomass obtained for characterization by HPLC. They were identified with molecular techniques and the CS13, P10A and P11A strains were made growth kinetics. The highest yield in pigment extraction was obtained by the CS13 strains with 10.2 µg / mL, the P10A strain with 9.7 µg / mL and the P11A strain with 7.5 µg / mL concentration for β-carotene. The kinetics of growth and pigment production during five days was optimal for the P11A strain, from 48 hours its concentration was 109.62 µg / mL and 1403.10 µg / mL at 120 hours of total carotenoids. In this study, it was found that yeasts isolated from aquatic systems are promising for the production of carotenoid pigments (including βcarotene), making their extraction and characterization viable for future biotechnological studies.
Identificación de péptidos aislados del caracol gigante africano (achatina fulica) y estudio de su actividad antimicrobiana
(Universidad Santiago de Cali, 2020) Martínez Esquivel, Richard Andrés; Varela Miranda,Rubén Eduardo; Oñate Garzon, Jose Fernando
Snails, due to their potential in bioprospecting, have been studied in different settings, one of which is obtaining peptides with therapeutic activity from African snail slime, since antitumor and cosmetic peptides have been identified in this species. The rapid spread of antibiotic-resistant microorganisms in recent decades and the poor development of new antimicrobial drugs has represented a significant public health problem worldwide. Therefore, it is necessary to look for alternatives for the design of new drugs, for example, antimicrobial peptides from snail slime type (PAMs). For the extraction of peptides from the African snail slime, the TRI Reagent®, reagent was used as a total protein extraction method, with a previous treatment with N-acetylcysteine for the elimination of the mucin present in the slime and thus improve the sample quality. Subsequently, the proteins extracted and filtered with AMICON were visualized with SDS gels with tricine at 14 and 16%, obtaining small bands mainly below 10 KDa, confirming the presence of peptides, despite this, the extract evaluated did not show antimicrobial activity. Finally, the data from the CIC (Cancer Research Center) of Spain also reported the isolation of peptides in the samples; however, its identification and synthesis are pending for the evaluation of its antimicrobial activity with pure peptides and in adequate concentrations. These results validate that the methodology used is appropriate for the isolation of peptides from the African snail slime in isolates from the city of Cali.
“Selección y caracterización de levaduras oleaginosas aisladas a partir de sistemas acuáticos de la ciudad de Cali”
(Universidad Santiago de Cali, 2020) Arcos Velasco, Lizeth Vanessa; Ramírez Castrillón, Mauricio; Osorio Cadavid, Esteban
Yeasts have been reported as a source of microbial oil of industrial interest, since the accumulation and lipid profile of each strain can present potential use as a raw material for the generation of biodiesel (monounsaturated fatty acids) or food (polyunsaturated fatty acids). Furthermore, oleaginous yeasts represent a promising option for this purpose, because they accumulate neutral lipids, mainly triglycerides, in a high proportion of biomass (20%-70%, w/w). So, the objective of this project was to evaluate the lipid accumulating capacity in a yeast collection previously isolated from aquatic systems in Cali, Colombia. 25 strains were analyzed to determine their possible oleaginous nature and a lipid production screening was performed by gravimetry (weight of total lipids with respect to their dry biomass), being selected the yeasts with higher percentage of lipids. These strains were identified at the species level using ribosomal regions sequencing and were compared with the sequences of type strains deposited in the Genbank and Mycobank databases. Five strains of yeasts with a yield of 20% g/g or more were selected for lipid production

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