Identification of Enzymatic Bottlenecks for the Aerobic Production of Malate from Glycerol by the Systematic Gene Overexpression of Anaplerotic Enzymes in Escherichia coli.

The biotechnological production of dicarboxylic acids (C4) from renewable carbon sources represents an attractive approach for the provision of these valuable compounds by green chemistry means. Glycerol has become a waste product of the biodiesel industry that serves as a highly reduced carbon source for some microorganisms. Escherichia coli is capable of consuming glycerol to produce succinate under anaerobic fermentation, but with the deletion of some tricarboxylic acid (TCA) cycle genes, it is also able to produce succinate and malate in aerobiosis. In this study, we investigate possible rate-limiting enzymes by overexpressing the C-feeding anaplerotic enzymes Ppc, MaeA, MaeB, and Pck in a mutant that lacks the succinate dehydrogenase (Sdh) enzyme. The overexpression of the TCA enzyme Mdh and the activation of the glyoxylate shunt was also examined. Using this unbiased approach, we found that phosphoenol pyruvate carboxylase (Ppc) overexpression enhances an oxidative pathway that leads to increasing succinate, while phosphoenol pyruvate carboxykinase (Pck) favors a more efficient reductive branch that produces mainly malate, at 57.5% of the theoretical maximum molar yield. The optimization of the culture medium revealed the importance of bicarbonate and pH in the production of malate. An additional mutation of the ppc gene highlights its central role in growth and C4 production.

Halotolerant Endophytic Bacteria Priestia flexa 7BS3110 with Hg2+ Tolerance Isolated from Avicennia germinans in a Caribbean Mangrove from Colombia.

The mangrove ecosystems of the Department of Atlántico (Colombian Caribbean) are seriously threatened by problems of hypersalinization and contamination, especially by heavy metals from the Magdalena River. The mangrove plants have developed various mechanisms to adapt to these stressful conditions, as well as the associated microbial populations that favor their growth. In the present work, the tolerance and detoxification capacity to heavy metals, especially to mercury, of a halotolerant endophytic bacterium isolated from the species Avicennia germinans located in the Balboa Swamp in the Department of Atlántico was characterized. Diverse microorganisms were isolated from superficially sterilized A. germinans leaves. Tolerance to NaCl was evaluated for each of the obtained isolates, and the most resistant was selected to assess its tolerance to Pb2+, Cu2+, Hg2+, Cr3+, Co2+, Ni2+, Zn2+, and Cd2+, many of which have been detected in high concentrations in the area of study. According to the ANI and AAI percentages, the most halotolerant strain was identified as Priestia flexa, named P. flexa 7BS3110, which was able to tolerate up to 12.5% (w/v) NaCl and presented a minimum inhibitory concentrations (MICs) of 0.25 mM for Hg, 10 mM for Pb, and 15 mM for Cr3+. The annotation of the P. flexa 7BS3110 genome revealed the presence of protein sequences associated with exopolysaccharide (EPS) production, thiol biosynthesis, specific proteins for chrome efflux, non-specific proteins for lead efflux, and processes associated with sulfur and iron homeostasis. Scanning electron microscopy (SEM) analysis showed morphological cellular changes and the transmission electron microscopy (TEM) showed an electrodense extracellular layer when exposed to 0.25 mM Hg2+. Due to the high tolerance of P. flexa 7BS3110 to Hg2+ and NaCl, its ability to grow when exposed to both stressors was tested, and it was able to thrive in the presence of 5% (w/v) NaCl and 0.25 mM of Hg2+. In addition, it was able to remove 98% of Hg2+ from the medium when exposed to a concentration of 14 mg/L of this metalloid. P. flexa 7BS3110 has the potential to bioremediate Hg2+ halophilic contaminated ecosystems.

Microbiological water quality and sources of contamination along the coast of the Department of Atlántico (Caribbean Sea of Colombia). Preliminary results.

Microbiological sea water quality is a public health problem that has serious repercussions in the tourism and economy of Colombia. This study determines the concentrations of Escherichia coli, Enterococcus faecalis and Clostridium perfringens at eleven beach water points and seven streams along the coast of the Department of Atlántico, Colombia. In seawater, total E. coli, E. faecalis and C. perfringens concentrations were found between 16 and 572 cfu/100 mL, 7-450 cfu/100 ml and 2-125 cfu/100 ml, respectively. The highest counts were observed mainly on urbanised beaches and in correspondence with streams whose waters had a high concentration of faecal origin microorganisms, which represent a serious health risk factor for bathers. Relevant efforts have to be addressed to improve the microbiological quality of these beaches by the establishment of efficient wastewater management programs aimed at enhancing the efficiency of the local treatment plant and the control of illegal sewage pouring onto the coast.

Molecular detection of Salmonella spp., Listeria spp. and Brucella spp. in fresh artisanal cheese marketed in the city of Barranquilla: A pilot study

Introduction: Each year approximately 3 million people die as the result of foodborne diseases. The fresh artisan (handmade) cheese produced and distributed in the Colombian Caribbean region is a native product from the departments of Córdoba, Sucre, Bolívar, Atlántico, Magdalena, Cesar, and La Guajira. Its mass consumption increases the risk of infection with Salmonella spp., Listeria spp., and Brucella spp., as it is made with a very rustic technology, with unpasteurized cow milk, without standardized and hygienic procedures and its storage is inadequate. Objective: To detect the presence of Salmonella spp., Listeria spp., and Brucella spp. in samples of fresh artisan cheese from the Colombian Caribbean region. Materials and methods: Twenty-seven samples of cheese from five departments of the Caribbean Region (Atlántico (n=6), Bolívar (n=2), Córdoba (n=1), Magdalena (n=16), and Sucre (n=2)) were analyzed by real-time polymerase chain reaction (qPCR). Seventeen of the samples corresponded to soft cheese, five to semi-hard cheese and five to hard cheese. Results: In 62.9% (17/27) of the samples we detected Salmonella spp., in 70.4% (19/27), Listeria spp., and in 22.2% (6/27), Brucella spp., mainly from the department of Magdalena. In 62.5% (10/16) of the samples we detected Salmonella spp. and Listeria spp. while in the department of Atlántico, 50% (3/6) of the samples corresponded to Brucella spp. Conclusion: The results confirmed the presence of these microorganisms in all the samples of soft cheese from the Colombian Caribbean region.

Detección molecular de Salmonella spp., Listeria spp. y Brucella spp. en queso artesanal fresco comercializado en Barranquilla: un estudio piloto / Molecular detection of Salmonella spp., Listeria spp. and Brucella spp. in fresh artisanal cheese marketed in the city of Barranquilla: A pilot study

Introducción. Cada año mueren, aproximadamente, tres millones de personas como consecuencia de las enfermedades transmitidas por alimentos. El queso artesanal fresco que se produce y distribuye en la región Caribe colombiana es un producto autóctono de los departamentos de Córdoba, Sucre, Bolívar, Atlántico, Magdalena, Cesar y La Guajira. Su consumo masivo aumenta el riesgo de infección con salmonelosis, listeriosis y brucelosis debido a que es elaborado con una tecnología muy rústica, con leche de vaca no pasteurizada, sin procedimientos estandarizados e higiénicos, y a que su almacenamiento no es adecuado. Objetivo. Detectar la presencia de Salmonella spp., Listeria spp. y Brucella spp. en muestras de queso artesanal costeño fresco procedente de los departamentos de la región Caribe colombiana. Materiales y métodos. Mediante reacción en cadena de la polimerasa en tiempo real (qPCR), se analizaron 27 muestras de queso proveniente de cinco departamentos de la región Caribe: Atlántico (n=6), Bolívar (n=2), Córdoba (n=1), Magdalena (n=16) y Sucre (n=2). Del total de las muestras, 17 eran de queso blando, cinco de queso semiduro y cinco de queso duro. Resultados. En el 62,9 % de las muestras se detectó Salmonella spp. (17/27), en el 70,4 %, Listeria spp. (19/27), y en el 22,2 %, Brucella spp. (6/27). Las muestras provenían principalmente del departamento del Magdalena y, en 62,5 % (10/16) de ellas, se encontró Salmonella spp. y Listeria spp., en tanto que en el 50 % (3/6) de las muestras del departamento del Atlántico se detectó Brucella spp. Conclusión. Los resultados evidenciaron la presencia de estos microorganismos en todas las muestras de queso costeño blando.

Introduction: Each year approximately 3 million people die as the result of foodborne diseases. The fresh artisan (handmade) cheese produced and distributed in the Colombian Caribbean region is a native product from the departments of Córdoba, Sucre, Bolívar, Atlántico, Magdalena, Cesar, and La Guajira. Its mass consumption increases the risk of infection with Salmonella spp., Listeria spp., and Brucella spp., as it is made with a very rustic technology, with unpasteurized cow milk, without standardized and hygienic procedures and its storage is inadequate. Objective: To detect the presence of Salmonella spp., Listeria spp., and Brucella spp. in samples of fresh artisan cheese from the Colombian Caribbean region. Materials and methods: Twenty-seven samples of cheese from five departments of the Caribbean Region (Atlántico (n=6), Bolívar (n=2), Córdoba (n=1), Magdalena (n=16), and Sucre (n=2)) were analyzed by real-time polymerase chain reaction (qPCR). Seventeen of the samples corresponded to soft cheese, five to semi-hard cheese and five to hard cheese. Results: In 62.9% (17/27) of the samples we detected Salmonella spp., in 70.4% (19/27), Listeria spp., and in 22.2% (6/27), Brucella spp., mainly from the department of Magdalena. In 62.5% (10/16) of the samples we detected Salmonella spp. and Listeria spp. while in the department of Atlántico, 50% (3/6) of the samples corresponded to Brucella spp. Conclusion: The results confirmed the presence of these microorganisms in all the samples of soft cheese from the Colombian Caribbean region.