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Biomedical subjects

F X Malcata

Publications and source records attributed to F X Malcata.

At least 19 recordsLinked to original sources
PubMed2006

Manufacturing of fermented goat milk with a mixed starter culture of Bifidobacterium animalis and Lactobacillus acidophilus in a controlled bioreactor.

AIMS: This work was undertaken to study the feasibility and the characteristics of a fermented product made of goat milk, using a mixed starter culture of Bifidobacterium animalis and Lactobacillus acidophilus under controlled conditions, and to determine their survival in the fermented milk during refrigerated storage. METHODS AND RESULTS: Goat milk was inoculated with Lact. acidophilus and Bif. animalis mixed starter, fermented in a glass bioreactor with controlled temperature (37 degrees C) and anaerobiosis, and monitored for growth and acidification. The fermented milk was then stored for 10 days under refrigeration, and monitored daily for starter microflora survival and pH changes. Lact. acidophilus viable counts reached a maximum of 7.1 x 10(8) colony-forming units (CFU) ml(-1), and Bif. animalis a maximum of 6.3 x 10(7) CFU ml(-1) by 20 h of fermentation. During refrigerated storage, both strains exhibited a good survival, with viable numbers remaining essentially constant throughout the experiment, whereas the pH of the fermented milk dropped slightly. CONCLUSIONS: Mixed cultures of Bif. animalis and Lact. acidophilus may be used to produce fermented goat milk with high counts of both probiotic strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Goat milk fermented with Bif. animalis and Lact. acidophilus can be manufactured as an alternative probiotic dairy product.

Animals↗
PubMed2006

Molecular characterization of peptides released from beta-lactoglobulin and alpha-lactalbumin via cardosins A and B.

Crude mixtures of aspartic proteases from flowers of the plant Cynara cardunculus have been studied frequently, as have activities of such enzymes (in pure form) on caseins from bovine, ovine, and caprine sources. This research study addressed pure bovine whey protein as substrates; that is, alpha-lactalbumin (alphaLA) and beta-lactoglobulin (alpha-LG), submitted to hydrolysis by 1 of 2 aspartic proteases (cardosins A and B), previously extracted and purified from C. cardunculus. Samples collected, following incubation at 55 degrees C and pH 5.2, were assayed by fast protein liquid chromatography, reversed phase-high performance liquid chromatography, and tricine-sodium dodecyl sulfate polyacrylamide gel electrophoresis; the major peptides released were then collected and sequenced by Edman degradation. Cardosin B and, to a lesser degree, cardosin A showed proteolytic activity toward alpha-LA, but the hydrolyzates produced were characterized by distinct peptide profiles. Cardosin B possesses a broad specificity, and produces several hydrophobic peptides (at least 5, with molecular mass in the range 2 to 8 kDa) in the early stages, which eventually become more hydrophilic (with molecular mass below 2 kDa) at later stages of hydrolysis. Cardosin A was found to cleave alpha-LA at the peptide bonds Phe28-Arg29, Gly54-Tyr55, Ala59-Ile60, Leu71-Phe72, and Leu105-Thr106, whereas cardosin B cleaved Ala19-Glu20, Phe28-Arg29, Glu30-Leu31, Tyr37-Gly38, Trp45-Val46, Phe50-His51, Ala59-Ile60, Ser66-Thr67, Leu71-Phe72, Phe72-Gln73, Gln73-Ile74, Ile78-Trp79, Leu115-Asp116, and Leu124-Ala125. Conversely, cardosins A and B are apparently not active on beta-LG.

Amino Acid Sequence↗
PubMed2006

Bioactive peptides in ovine and caprine cheeselike systems prepared with proteases from Cynara cardunculus.

The potential angiotensin-converting enzyme (ACE)-inhibitory and antioxidant activities of peptides in water-soluble extracts, obtained from raw and sterilized ovine and caprine cheeselike systems coagulated with enzymes from the plant Cynara cardunculus, were assessed. Prior to the assay, the 3,000-Da permeate from 45-d-old cheeselike systems was fractionated by tandem chromatographic techniques. Several peaks were obtained in each chromatogram, but only some were associated with ACE-inhibitory or antioxidant activity or both. Peptides Tyr-Gln-Glu-Pro, Val-Pro-Lys-Val-Lys, and Tyr-Gln-Glu-Pro-Val-Leu-Gly-Pro-* from beta-casein, as well as Arg-Pro-Lys and Arg-Pro-Lys-His-Pro-Ile-Lys-His-* from alpha(s1)-casein exhibited ACE-inhibitory activity. Peptides released upon cleavage of the peptide bond Leu190-Tyr191 (either in ovine or caprine beta-casein), and corresponding to the beta-casein sequence Tyr-Gln-Glu-Pro-*, possessed antioxidant activity.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed2006

Detection and characterization of Listeria monocytogenes in Sao Jorge (Portugal) cheese production.

Listeria monocytogenes is a foodborne pathogen that can cause serious invasive disease in humans. Because human listeriosis cases have previously been linked to consumption of contaminated cheese, control of this pathogen throughout the cheese production chain is of particular concern. To understand the potential for L. monocytogenes transmission via São Jorge cheese, a Portuguese artisanal cheese variety that bears a Protected Denomination of Origin classification, 357 raw milk, curd, natural whey starter, and cheese samples representative of the production chain of this cheese were collected over one year and tested for the presence of L. monocytogenes and selected physicochemical parameters. Although neither L. monocytogenes nor other Listeria spp. were detected in whey, curd, or cheese samples, 2 of the 105 raw milk samples analyzed were positive for L. monocytogenes. These 2 raw milk isolates represented a ribotype that has previously been linked to multiple human listeriosis outbreaks and cases elsewhere, indicating the potential of these isolates to cause human listeriosis. On average, physicochemical parameters of São Jorge cheese ripened for 4 mo presented values that likely minimize the risk of L. monocytogenes outgrowth during ripening and storage (mean pH = 5.48; mean moisture = 37.79%; mean NaCl concentration = 4.73%). However, some cheese samples evaluated in this study were characterized by physicochemical parameters that may allow growth and survival of L. monocytogenes. Even though our results indicate that raw milk used for São Jorge cheese manufacture as well as finished products is rarely contaminated with L. monocytogenes, continued efforts to control the presence of this pathogen in the São Jorge cheese production chain are urged and are critical to ensure the safety of this product.

Animals↗
PubMed2005

Partial identification of water-soluble peptides released at early stages of proteolysis in sterilized ovine cheese-like systems: influence of type of coagulant and starter.

Cheese-like systems were manufactured from sterilized ovine milk, using crude aqueous extracts of Cynara cardunculus or cardosin A isolated therefrom as clotting agent. The effect of adding a commercial starter culture was also assessed. The impact of the type of coagulant used during the initial 24 h of proteolysis was evaluated via separation of peptides in the water-soluble extracts by reverse-phase HPLC, followed by partial sequencing via Edman degradation. Cardosin A accounted for most events of primary proteolysis. The major cleavage sites were Phe105-Met106 in kappa-casein, and Leu127-Thr128, Ser142-Trp143, Leu165-Ser166, and Leu190-Tyr191 in beta-casein. The starter culture did not play an active role during the initial stages of ripening.

Animals↗
PubMed2003

Effect of various growth media upon survival during storage of freeze-dried Enterococcus faecalis and Enterococcus durans.

AIMS: The effects of three different growth media (MRS, M17 and Lee's) on survival during freeze-drying and subsequent storage of six strains of Enterococcus faecalis and two strains of E. durans were investigated. METHODS AND RESULTS: Distinct Enterococcus spp. strains were grown on M17, MRS and Lee's broth, freeze-dried and stored at 20 degrees C in air under darkness. At regular intervals throughout storage, freeze-dried samples were rehydrated and then plated on M17 agar. CONCLUSIONS: A higher survival rate during storage of dried E. durans was obtained when growth occurred in MRS. The same effect was not observed, however, for the majority of E. faecalis strains, which clearly survived better in the dried state when this organism had been grown in M17 or Lee's medium. SIGNIFICANCE AND IMPACT OF STUDY: The survival of the dried Enterococcus spp. tested during storage was shown to be strain-specific and dependent on the growth medium.

Culture Media↗
PubMed2002

Amino acid catabolism and generation of volatiles by lactic acid bacteria.

Twelve isolates of lactic acid bacteria, belonging to the Lactobacillus, Lactococcus, Leuconostoc, and Enterococcus genera, were previously isolated from 180-d-old Serra da Estrela cheese, a traditional Portuguese cheese manufactured from raw milk and coagulated with a plant rennet. These isolates were subsequently tested for their ability to catabolize free amino acids, when incubated independently with each amino acid in free form or with a mixture thereof. Attempts were made in both situations to correlate the rates of free amino acid uptake with the numbers of viable cells. When incubated individually, leucine, valine, glycine, aspartic acid, serine, threonine, lysine, glutamic acid, and alanine were degraded by all strains considered; arginine tended to build up, probably because of transamination of other amino acids. When incubated together, the degradation of free amino acids by each strain was dependent on pH (with an optimum pH around 6.0). The volatiles detected in ripened Serra da Estrela cheese originated mainly from leucine, phenylalanine, alanine, and valine, whereas in vitro they originated mainly from valine, phenylalanine, serine, leucine, alanine, and threonine. The wild strains tested offer a great potential for flavor generation, which might justify their inclusion in a tentative starter/nonstarter culture for that and similar cheeses.

Amino Acids↗
PubMed2001

Decolourisation of Remazol Brilliant Blue R via a novel Bjerkandera sp. strain.

A novel strain of Bjerkandera sp. (B33/3), with particularly high decolourisation activities upon Poly R-478 and Remazol Brilliant Blue R (RBBR) dyes, was isolated. The role of the ligninolytic extracellular enzymes produced by this strain on decolourisation of RBBR was studied in some depth. The basis of decolourisation is an enzyme-mediated process, in which the main enzyme responsible is a recently described peroxidase with capacity for oxidation of manganese, as well as veratryl alcohol and 2,6-dimethoxyphenol in a manganese-independent reaction.

Anthraquinones↗
PubMed2001

Activity of adventitious Enterococcus strains on model curdled caprine milk: microbial growth and evolution of concentration of organic acids and lactose throughout time.

Enterococci offer a few unique advantages when compared with other lactic acid bacteria in that they are resistant to phages, stable lactose fermenters, tolerant to relatively high concentrations of sodium chloride and optimal growers at relatively high temperatures (Franz et al., 1999). Such genus occurs and grows in a variety of cheeses, especially artisanal cheeses produced in Southern Europe usually from raw milk; they are claimed to play a major role in ripening and aroma development in such cheeses (Freitas et al., 1995; Macedo et al., 1995). Enterococci can produce lactic, acetic, propionic and succinic acids (Ocando et al., 1994), compounds that have been correlated with flavor characteristics of cheese; hence strains of Enterococcus have been successfully used as starters for different cheese (Franz et al., 1999).

Animals↗
PubMed2000

Microbiology and biochemistry of cheeses with Appélation d'Origine Protegée and manufactured in the Iberian Peninsula from ovine and caprine milks.

To support legal protection with objective technical data and to promote enforcement of high quality standards a few European countries have created Appélation d'Origine Protegées. This paper reviews and updates fundamental and applied aspects encompassing microbiological and biochemical characteristics of traditional cheeses with Appélation d'Origine Protegée manufactured in the Iberian Peninsula from ovine, caprine, or both milks. Ovine and caprine cheeses with Appélation d'Origine Protegée from Portugal and Spain can be divided into four distinct groups based on milk source and rennet type: 1) Azeitão, Castelo Branco, Evora, Nisa, Serpa, Serra da Estrela, and La Serena cheeses are manufactured with raw ovine milk and coagulated via plant rennet; 2) Terrincho, Idiazábal, Manchego, Roncal, and Zamorano cheeses are manufactured with raw ovine milk and coagulated via animal rennet; 3) Cabra Transmontano and Majorero are manufactured with raw caprine milk and coagulated via animal rennet; and 4) Amarelo da Beira Baixa, Picante da Beira Baixa, and Rabaçal are manufactured with mixtures of raw ovine and caprine milks and coagulated via animal rennet.

Animals↗
PubMed2000

Characterization of whey cheese packaged under vacuum.

Vacuum packaging was assayed at 4 degrees C and was tested in comparison to unpackaged counterparts, in both microbiological and physicochemical terms, in studies pertaining to the preservation of Requeijão, a traditional Portuguese whey cheese. Bacteria were absent (i.e., <10 CFU/g) in whey cheeses on the day of manufacture as a result of thermal processing. After storage, both unpackaged and packaged cheeses exhibited high viable counts of Bacillus, Pseudomonas, Enterobacteriaceae, and lactic acid bacteria (especially lactococci). Yeasts, staphylococci, enterococci, and spore-forming clostridia were severely inhibited by the package vacuum combined with the increasing acidification developed therein. Whey cheeses packaged under vacuum underwent substantial acidification, slight depletion of lactose, and no significant variation in moisture content or texture; conversely, unpackaged whey cheeses exhibited substantial loss of water and a concomitant increase in rigidity. Vacuum packaging strongly inhibited lipolysis (even if viable counts of some microbial groups were high); saturated fatty acids (mainly C16:0 and C14:0) accounted for ca. 73% of the total free-fatty acid content, whereas the most concentrated unsaturated fatty acids were C18:1 and C18:2 (ca. 14% each). The conclusions generated in our study are, in general, useful for a wide range of whey cheeses worldwide: i.e., Requéson (Spain), Ricotta (Italy), Broccio (France), and Anthotyro (Greece). In addition, our conclusions are particularly helpful in terms of improving the safety of Requeijão, a widely acclaimed dairy specialty.

Cheese↗
PubMed1999

ADSA Foundation Scholar Award. Critical issues affecting the future of dairy industry: individual contributions in the scope of a global approach.

Several constraints that have been affecting the dairy industry are identified in a critical fashion, and directions are given with an emphasis on food processing implemented at the postproduction level. The rationale for modifications aimed at enhancing the appeal of condensed dairy products should be consubstantiated in strengthening of organoleptic characteristics, improvement of nutraceutical impact, and reduction of polluting power. This enumeration follows an order of increasing time scale required for consumer perception and increasing size scale associated with expected impact. Pursuance of such streamlines should lead to manufacture of dairy products that resemble nature more closely in terms of milk coagulation, milk fat modification, milk fermentation, whey fermentation, and starter culture addition. Directions for research and development anticipated as useful and effective in this endeavor, and which have been previously and consistently adopted in the development of an individual research program, are characterization and development of alternative rennets from plant sources, development of starter and nonstarter cultures from adventitious microflora, utilization of probiotic strains as starter cultures, upgrading of whey via physical or fermentation routes, and modification of milk fat via lipase-mediated interesterification reactions.

Animals↗
PubMed1999

On the microbiological profile of traditional Portuguese sourdough.

Traditional manufacture of bread from maize has been noted to play important roles from both economic and social standpoints; however, enforcement of increasingly strict hygiene standards requires thorough knowledge of the adventitious microbiota of the departing dough. To this goal, sourdough as well as maize and rye flours from several geographic locations and in two different periods within the agricultural year were assayed for their microbiota in sequential steps of quantification and identification. More than 400 strains were isolated and taxonomic differentiation between them was via Biomerieux API galleries (375 of which were successfully identified) following preliminary biochemical and morphological screening. The dominant groups were yeasts and lactic acid bacteria (LAB). The most frequently isolated yeasts were Saccharomyces cerevisiae and Candida pelliculosa. The most frequently isolated LAB were (heterofermentative) Leuconostoc spp. and (homofermentative) Lactobacillus spp.; L. brevis, L. curvatus, and L. lactis ssp. lactis were the dominant species for the Lactobacillus genera; Lactococcus lactis ssp. lactis for lactococci; Enterococcus casseliflavus, E. durans, and E. faecium for enterococci; and Streptococcus constellantus and S. equinus for streptococci.

Bacteria↗
PubMed1998

Lipase catalyzed modification of milkfat.

Decreasing consumption of high fat milk and dairy products is driving the dairy industry to seek other uses for increasing surplus of milkfat. Enzyme catalyzed modification of milkfat using lipases is receiving particular attention. This review examines lipase-mediated modification of milkfat. Especial attention is given to industrial applications of lipases for producing structured and modified milkfat for improved physical properties and digestibility, reduced caloric value, and flavor enhancement. Features associated with reactions such as hydrolysis, transesterification, alcoholysis and acidolysis are presented with emphasis on industrial feasibility, marketability and environmental concerns. Future prospects for enzyme catalyzed modification of milk fat are discussed.

Journal Article↗
PubMed1998

The microstructure and distribution of micro-organisms within mature Serra cheese.

The distribution of micro-organisms in mature Serra, a traditional Portuguese cheese made from unpasteurised ewes' milk without added starter culture, was examined by light microscopy and electron microscopy. Four populations of micro-organisms were recognized according to their position within the cheese: (i) those present as apparently axenic colonies within the curd matrix; (ii) bacteria growing along curd junctions; (iii) yeasts and bacteria present in the smear on the surface of the cheese and (iv) bacteria found in cracks which penetrated the outer part of the cheese from the rind. Two types of crystals were observed, together with contaminants of vegetable origin and somatic cells originating from the milk.

Animals↗
PubMed1998

Use of small ruminants' milk supplemented with available nitrogen as growth media for Bifidobacterium lactis and Lactobacillus acidophilus.

Growth of, and acid production by Bifidobacterium lactis and Lactobacillus acidophilus using ovine and caprine milk as media were evaluated for their potential use in cheese-making. A protein hydrolysate (MHP, obtained from incubation of bovine milk with protease) or a mixture of free amino acids (FAA, similar to the amino acid fraction of MHP) was added as a nitrogen enrichment source. Bifidobacterium lactis and Lact. acidophilus were inoculated at 50 ml l-1 and incubated at 37 degrees C with growth supplements added at ratios in the range 25-50 ml l-1. The maximum viable counts of Bif. lactis were lower in plain ovine and caprine milk than in nitrogen-enriched milk, and MHP was a better growth promoter than FAA. A similar trend was observed with the acidity values developed, and attempts to correlate growth with acidity were successfully performed. The highest uptake rates of amino acids in ovine milk were observed for lysine, isoleucine, leucine and proline, but only isoleucine was taken up at a similar rate in caprine milk. Final bacterial viable counts of Lact. acidophilus in the plain and enriched forms of ovine milk did not differ greatly from each other, although FAA was statistically a better growth promoter than MHP. Unlike results in ovine milk, cultures of Lact. acidophilus in caprine milk exhibited drops of 1-1.5 log cycles in viable cell counts by 24 h of fermentation, irrespective of the nature of the nitrogen source. Parallel studies indicated that the excess of fatty acid residues in caprine milk could be responsible for the poor growth of Lact. acidophilus.

Amino Acids↗