Humberto Hernández-Sánchez

Antibacterial activity of Capsicum extract against Salmonella typhimurium and Pseudomonas aeruginosa inoculated in raw beef meat

The inhibitory effect of the extract of Capsicum annuum bell pepper type was evaluated against Salmonella typhimurium and Pseudomonas aeruginosa, inoculated in minced beef meat mixed with different concentrations of the extract, and stored at 7 degrees C for 7 days. The combined effect of C. annuum extract and sodium chloride on the bacterial growth was evaluated. The minimum inhibitory concentration of the extract to prevent the growth of S. typhimurium in minced beef was 1.5 ml/100 g of meat; the addition of 1%, 2%, 3% and 4% w/w of sodium chloride did not have any additional inhibitory effect on Salmonella. In the case of P. aeruginosa, a concentration of 0.3 ml of the extract/100 g of meat showed a bacteriostatic effect, while a concentration of 3 ml/100 g of meat showed a bactericidal effect. When 1% w/w of sodium chloride was added to the meat together with the extract, the concentration needed to kill P. aeruginosa was reduced.

Effect of Thermal Treatment on the Antioxidant Activity and Content of Carotenoids and Phenolic Compounds of Cactus Pear Cladodes (Opuntia ficus-indica)

Cactus pears (Opuntia ficus-indica) are draught resistant plants originated in Mexico. Their flattened stem segments, called cladodes, have moisture, protein and fibre contents of 92, 1-2 and 4-6% respectively, and a pectin content in the range of 0.8-3.3% depending on the species. They also contain certain concentration of carotenoids which are of special interest because of their antioxidant activity. This work is aimed to identify and quantify the main carotenoids present in the stems and to evaluate the effect of thermal treatments on the antioxidant activity and concentration of carotenoids and phenolic compounds. The carotenoids -cryptoxanthin, -carotene and lutein were identified in the cladodes, the latter having the highest concentration. Thermal treatments increased the extractability of these pigments and the antioxidant activity was related to the carotenoids concentration. Total phenolic content decreased after the thermal treatments; however this result had little effect on the antioxidant activity. Mucilage present in the stems decreased the extractability of the carotenoids.

Behaviour of Listeria monocytogenes during the manufacture and ripening of Manchego and Chihuahua Mexican cheeses

The ability of Listeria monocytogenes to survive the Mexican Manchego and Chihuahua cheese-making processes and its persistence during the ripening stages of both cheeses was examined. Commercial pasteurized and homogenized whole milk was inoculated with Listeria monocytogenes (strain ATCC 19114) to a level between 2 x 10(6) and 9 x 10(6) CFU/ml. The milk was used to make Mexican Manchego and Chihuahua cheeses in a 25-l vat. Mexican Manchego cheese was ripened for 5 days and Chihuahua cheese for 6 weeks at 12 degrees C and 85% RH. Listeria present in the cheese was enumerated by diluting samples in sterile 0.1% peptone water and plating on Oxford agar. Duplicate samples were taken at each step of the manufacturing process. During the first week of ripening samples were taken daily from both cheeses. For Chihuahua cheese, samples were taken weekly after the first week of the ripening stage. During the manufacture of Mexican Manchego cheese, Listeria counts remained relatively constant at 10(6) CFU/ml, while with Chihuahua cheese there was a one log decrease in numbers (10(6) to 10(5) CFU/ml). After pressing both curds overnight, numbers of bacteria decreased in Mexican Manchego cheese to 8.2 x 10(5) but increased in Chihuahua cheese from 1.7 x 10(5) to 1.2 x 10(6) CFU/ml. During the ripening stage, counts of Listeria remained constant in both cheeses. However, since the Chihuahua cheese ripening stage is about 6 weeks, the number of bacteria decreased from 2 x 10(6) to 4 x 10(4) CFU/g. The results show that Listeria monocytogenes is able to survive the manufacture and ripening processes of both Mexican cheeses.

Effect of different treatments on the ability of α-lactalbumin to form nanoparticles

Nanoparticles of bovine α-lactalbumin (α-LA) prepared by desolvation and glutaraldehyde crosslinking are promising carriers for bioactive compounds in foods. The objective of this work was to study the effect of changes in hydrophobic interactions by using different desolvating agents (acetone, ethanol, or isopropanol) and the use of a heat or high-pressure treatment step before the desolvation process on the size, structure, and properties of α-LA nanoparticles. In all cases, a high average particle yield of 99.63% was obtained. Smaller sizes (152.3 nm) can be obtained with the use of acetone as the desolvating agent and without any pretreatment. This is the first time that α-LA nanoparticles in the size range of 100 to 200 nm have been obtained. These nanoparticles, with an isoelectric point of 3.61, are very stable at pH values >4.8, based on their ζ-potential, although their antioxidant activity is weak. The use of the desolvating agent with the smallest polarity index (isopropanol) produced the largest particles (293.4 to 324.9 nm) in all cases. These results support the idea that controlling hydrophobic interactions is a means to control the size of α-LA nanoparticles. No effect of pretreatment on nanoparticle size could be detected. All types of nanoparticles were easily degraded by the proteolytic enzymes assayed.

Isolation and Partial Characterization of Halotolerant Lactic Acid Bacteria from Two Mexican Cheeses

Isolated strains of halotolerant or halophilic lactic acid bacteria (HALAB) from Cotija and doble crema cheeses were identified and partially characterized by phenotypic and genotypic methods, and their technological abilities were studied in order to test their potential use as dairy starter components. Humidity, aw, pH, and salt concentration of cheeses were determined. Genotypic diversity was evaluated by randomly amplified polymorphic DNA-polymerase chain reaction. Molecular identification and phylogenetic reconstructions based on 16S rRNA gene sequences were performed. Additional technological abilities such as salt tolerance, acidifying, and proteolytic and lipolytic activities were also investigated. The differences among strains reflected the biodiversity of HALAB in both types of cheeses. Lactobacillus acidipiscis, Tetragenococcus halophilus, Weissella thailandensis, and Lactobacillus pentosus from Cotija cheese, and L. acidipiscis, Enterococcus faecium, Lactobacillus plantarum, Lactobacillus farciminis, and Lactobacillus rhamnosus from doble crema cheese were identified based on 16S rRNA. Quantitative and qualitative assessments showed strains of T. halophilus and L. plantarum to be proteolytic, along with E. faecium, L. farciminis, and L. pentosus to a lesser extent. Lipolytic activity could be demonstrated in strains of E. faecium, L. pentosus, L. plantarum, and T. halophilus. Strains belonging to the species L. pentosus, L. plantarum, and E. faecium were able to acidify the milk media. This study evidences the presence of HALAB that may play a role in the ripening of cheeses.

Evaluation of Phenylpropanoids in Ten Capsicum annuum L. Varieties and Their Inhibitory Effects on Listeria monocytogenes Murray, Webb and Swann Scott A

Extracts of pepper from ten different varieties of Capsicum annuumL. were prepared and their phenylpropanoids content was analysed by HPLC. There were seven different compounds identified at varying concentrations depending on pepper variety (L-phenylalanine, t-cinnamic acid, o-coumaric acid, mcoumaric acid, ferulic acid, caffeic acid and capsaicin) which are intermediates of the capsaicinoids pathway. The inhibitory action of the pepper extracts was evaluated against Listeria monocytogenes Murray, Webb and Swann Scott A by the agar diffusion test. Results showed that three of the 10 extracts had an antibacterial effect. Guajillo San Luis pepper presented the highest inhibitory effect, and contained the major concentration of cinnamic and caffeic acids. In order to analyse the individual contribution of every phenylpropanoid to the bacteriostatic effect, different concentrations were tested on the growth and survival of L. monocytogenesSwann Scott A.There were no significant differences (p> 0.05) in the t-cinnamic, o-coumaric, m-coumaric, ferulic acids and capsaicin among the samples. However, a significant difference was found between these compounds and the bactericidal action of caffeic acid. The results confirmed that L. monocytogenesSwann Scott A is inhibited by some pepper extracts, and also that some specific phenylpropanoids had a bacteriostatic effect.

Effects of high hydrostatic pressure on the structure of bovine α-lactalbumin

The effects of high hydrostatic pressure (HHP) processing (at 200 to 600 MPa, 25 to 55 degrees C, and from 5 to 15 min) on some structural properties of alpha-lactalbumin was studied in a pH range of 3.0 to 9.0. The range of HHP processes produced a variety of molten globules with differences in their surface hydrophobicity and secondary and tertiary structures. At pH values of 3 and 5, there was a decrease in the alpha-helix content concomitant with an increase in beta-strand content as the pressure increased. No changes in molecular size due to HHP-induced aggregation were detected by sodium dodecyl sulfate-PAGE. All samples showed higher thermostability as the severity of the treatment increased, indicating the formation of a less labile structure related to the HHP treatment.

Improvement of Fermentation Conditions for the Production of X-Prolyl-Dipeptidyl Aminopeptidase from Lactococcus Lactis

The quantitative effects of some fermentation conditions on the production of the enzyme X-prolyl-dipeptidyl aminopeptidase (PepXP)(EC 3.4.14.5) of Lactococcus lactis subsp. lactis and cremoris were studied. The PepXP activity was found both in the membrane and in the cytoplasm, suggesting the presence of multiple molecular forms. Both microorganisms showed higher PepXP activities when glucose (5 g/l) was used as the carbon source and the yeast extract in the culture medium was increased to 3.5 g/l. In these conditions, 226 mU/ml of PepXP activity were obtained with L. lactis subsp. lactis and 235 mU/ml with the subsp. cremoris after 6 h. The best fermentation temperature was in the 30–32 °C range. The enzyme activity remained stable even during the stationary phase.

Commercial probiotic bacteria and prebiotic carbohydrates: a fundamental study on prebiotics uptake, antimicrobials production and inhibition of pathogens.

BACKGROUND Probiotics and prebiotics are among the most important functional food ingredients worldwide. The proven benefits of such ingredients to human health have encouraged the development of functional foods containing both probiotics and prebiotics. In this work, the production of antimicrobial compounds coupled to the uptake of commercial prebiotics by probiotic bacteria was investigated. RESULTS The probiotic bacteria studied were able to take up commercial prebiotic carbohydrates to the same or higher extent than that observed for lactose (control carbohydrate). The growth of probiotic bacteria was coupled to the production of antimicrobials such as short-chain fatty acids (SCFA), H2 O2 and bacteriocins. A higher production of antimicrobial compounds was recorded with Oligomate 55® compared with Regulact® and Frutafit® (3-5 and 10-115 times higher SCFA and H2 O2 production, respectively). The probiotic bacteria grown with Oligomate 55® also produced bacteriocins and other non-identified antimicrobial compounds. The antimicrobials produced by the probiotic bacteria inhibited up to 50% the growth of model pathogens such as Escherichia coli, Listeria innocua and Micrococcus luteus compared with control cultures. CONCLUSIONS The results here obtained are useful for the adequate selection of probiotic/prebiotics pairs and therefore in the development of efficient functional foods.

α-Lactalbumin nanoparticles prepared by desolvation and cross-linking: structure and stability of the assembled protein.

A key step in the preparation of cross-linked protein nanoparticles involves the desolvation of proteins with an organic solvent, which is thought to act by modulating hydrophobic interactions. However, to date, no study has examined the conformational changes that proteins undergo during the assembly process. In this work, by using several biophysical techniques (CD spectroscopy, DSC, TEM, etc.), we studied spheroidal nanoparticles made from bovine α-lactalbumin cross-linked with glutaraldehyde in the presence of acetone. Within the nanoparticle, the polypeptide chain acquires a β-strand-like conformation (completely different from the native protein in secondary and tertiary structure) in which several side chains likely become available for reacting with glutaraldehyde. A multiplicity of cross-linking sites, together with the polymeric nature of glutaraldehyde, may thus explain the low dry-weight fraction of protein that was found in the nanoparticles. Although covalent bonds undoubtedly constitute the main source for nanoparticle stability, noncovalent interactions also appear to play a role in this regard.