[Show abstract][Hide abstract] ABSTRACT: The antimicrobial activity of the hydro-alcohol extract of Myrtus communis L. (ME) berries was investigated against six Listeria monocytogenes strains (2 type strains and 4 isolates). Sub-lethal ME concentrations reduced L. monocytogenes counts by at least 2 log cycles. A Central Composite Design was used to investigate the combined effects of sub-lethal concentrations of ME (0.039–0.195 mL/100 mL), NaCl (0–2.0 g/100 mL) and pH (5.0–7.0) on strains growth. ME affected growth parameters, generally extending lag phase length and reducing maximum growth, sometimes with interactive effects with pH. The highest ME concentrations (0.117–0.195 mL/100 mL) combined with the lowest pH values (5.0–6.0) strongly reduced or even inhibited strains growth.
Total phenolic content (TPC) and radical scavenging activity were also determined. ME had a TPC of 5315 ± 20 mg/kg of gallic acid equivalent, and malvidin-3-O-glucoside was the most abundant anthocyanin. In vitro radical scavenging activity, determined by TEAC assay, was stable during 70 days of refrigerated storage.
In conclusion, low ME concentrations, combined with salt and pH, were effective in reducing or containing cell growth. This antilisterial effect has interesting perspectives on industrial application; ME hydro-solubility can be considered a useful feature when antilisterial strategies are applied in food formulation.
LWT - Food Science and Technology. 01/2014; 58(1):116–123.
[Show abstract][Hide abstract] ABSTRACT: Aim of the work was to identify sulfide-producers isolated from tuna and swordfish and to evaluate some physiological characteristics, particularly those that could be related to spoiling potential.
16S rDNA sequencing revealed Shewanella baltica as the main species, followed by Serratia spp. and other Shewanella species, while RAPD- and rep-PCR analyses indicated the presence of several biotypes. Shewanella baltica and Shewanella putrefaciens showed rapid growth at 4 and 8 °C, production of TMA and H2S, amino acids decarboxylation and proteolytic activity also at refrigeration temperatures, therefore being potentially able to modify texture and sensory characteristics of finfish. Extracellular DNAse activity and growth in presence of high salt concentrations can provide a competitive advantage in unfavourable environments.
Our data provide new insights into specific metabolic features of Shewanella spp., rarely studied before, such as extracellular DNAse activity and amino acid decarboxylating activity. Moreover, our results highlight the clear necessity of more specific media and research methods to count H2S-producing bacteria.
[Show abstract][Hide abstract] ABSTRACT: CAMP test reliably detects Listeria monocytogenes (Lm) and Streptococcus agalactiae (group B streptococcus, GBS); it is traditionally performed streaking the tested isolate perpendicularly to Staphylococcus aureus (Sa), provided that reference Sa strains (that produce β-hemolysin) are used. In a zone of β-hemolysin activity, in fact, GBS and Lm form typical arrow-shaped hemolytic areas. While Sa production of the toxin is strain-dependent, however, that of Staphylococcus pseudintermedius (Sp), a pet-owner colonizer and an emerging human pathogen, is constitutive, then observed in all clinical isolates. Therefore, Sp may indeed represent a valid alternative to perform the assay.
International journal of clinical and experimental pathology 01/2014; 7(4):1733-4. · 2.24 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This study aimed at evaluating co-cultures of selected microorganisms for their proteolytic activity and capability to produce fermented milk enriched with ACE-inhibitory (ACEI) peptides. Selected yeasts (Torulaspora delbruekii KL66A, Galactomyces geotrichum KL20B, Pichia kudriavzevii KL84A and Kluyveromyces marxianus KL26A) and lactic acid bacteria strains (Lactobacillus plantarum LAT03, Lb. plantarum KLAT01 and the not virulent Enterococcus faecalis KE06) were screened as single cultures for their capacity of releasing ACEI peptides without producing bitter taste. Three strains cultures (yeast, Lb. plantarum and E. faecalis) were performed to evaluate the combined impact on microbial growth, lactic acid production, citric acid consumption, proteolysis, ACEI activity, and bitter taste after 36 h of fermentation at 28 °C. While G. geotrichum KL20B showed a strong stimulating effect on Lb. plantarum strains and the production of peptides with ACEI activity, the presence of T. delbruekii KL26A in the cultures was deleterious both to ACEI activity and product taste. The most effective combination was P. kudriavzevii KL84A, Lb. plantarum LAT3, E. faecalis KL06, which showed the highest ACEI activity (IC50 = 30.63 ± 1.11 μg ml−1) and gave no bitter taste for 7 days at 6 °C. Our results highlight the importance of choosing the strains combination carefully, to obtain a high yield of ACEI activity without bitter taste.
[Show abstract][Hide abstract] ABSTRACT: Listeria monocytogenes infection in pregnant women and newborns is a cause for serious concern, and invasive disease outcome strongly depends on prompt antibiotic therapy. To provide sooner identification from neonatal bacteremia we performed a CAMP test directly on positive blood aliquots and inoculated the Liofilchem(®) O.A. Listeria chromogenic agar as well, thus providing a 24-h turn-around time for response.
International journal of clinical and experimental pathology 01/2014; 7(3):1172-5. · 2.24 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This study investigated the possibility of using yeast strains in fermented milks to obtain products with high Angiotensin I-converting enzyme (ACE) inhibitory activity and low bitter taste. Ninety-three yeast strains isolated from Colombian Kumis in different geographic regions were molecularly identified, and their milk fermentation performances were determined. Molecular identification evidenced that Galactomyces geotrichum, Pichia kudriavzevii, Clavispora lusitaniae and Candida tropicalis, were the dominant species. Eighteen out of 93 strains produced fermented milk with ACE-inhibitory (ACEI) activity values ranging from 8.69 to 88.19%. Digestion of fermented milk samples by pepsin and pancreatin demonstrated an increase in ACEI activity, with C. lusitaniae KL4A as the best producer of ACEI peptides. Moreover, sensory analysis of the products containing the major ACE-inhibitory activity pointed out that P. kudriavzevii KL84A and Kluyveromyces marxianus KL26A could be selected as potential adjunct starter cultures in Kumis, since they made a considerable contribution to the ACE inhibitory activity and produced fermented milk without bitter taste. In this study we observed that Colombian Kumis can be an excellent vehicle for the isolation of yeasts with a potential to enhance bioactive peptides produced during milk fermentation.
International journal of food microbiology 08/2012; 159(1):39-46. · 3.01 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Kumis is a traditional fermented cow milk produced and consumed in South West Colombia. The main objective of this research was to studied the enterococcal population, present in 13 kumis samples traditionally manufactured, for their role as beneficial organisms or opportunistic pathogens. The molecular identification of 72 isolates evidenced that Enterococcus faecalis and E. faecium were the dominant species. The genes gelE, esp, asa1, cyl and hyl, all associated with virulence factors in enterococci, were detected in 30 isolates, while 42 were free of virulence determinants. Skim milk media were fermented by all the different isolates and further tested for proteolysis (free NH(3) groups), Angiotensin-I Converting Enzyme (ACE) inhibitory activity and biogenic amines production. Nine E. faecalis and two E. faecium strains produced fermented milk with ACE-inhibitory activity values ranging from 39.7% to 84.35% .The digestion of fermented milk samples by pepsin and pancreatin evidenced an increase in ACE inhibitory activity, with E. faecalis KE09 as the best producer (IC50 = 14.25 μg ml(-1)). Moreover, the strains showed a very low tyrosine decarboxylase activity and did not produce histamine during 48 h fermentation in milk. This study underlines the that Colombian kumis is a good source of not virulent enterococci able to produce fermented milks with ACE-inhibitory activity.
[Show abstract][Hide abstract] ABSTRACT: Strains of Saccharomyces cerevisiae isolated from Italian salami were screened for proteolytic activity in a model system containing sarcoplasmic (SMS) or myofibrillar (MMS) proteins, at 20°C for 14days, to evaluate the possible influence on the proteolysis of fermented sausages. SDS-PAGE revealed that 14 of the most osmotolerant strains were responsible for the extensive hydrolysis of the main myofibrillar proteins, while only one strain was able to hydrolyze sarcoplasmic proteins. Free amino acids (FAA) accumulated during proteolysis were strain-dependent with different patterns from sarcoplasmic or myofibrillar protein fraction. In general, proteolysis lead Cys, Glu, Lys and Val as the most abundant FAA in the inoculated MMS samples. Volatile compound analysis, determined by SPME-GC-MS, evidenced 3-methyl butanol in MMS, and 2-methyl propanol and 3-methyl-1-butanol in SMS as major compounds. Our findings highlight that S. cerevisiae could influence the composition in amino acids and volatile compounds in fermented sausages, with a strain-dependent activity.
International journal of food microbiology 07/2011; 150(1):50-8. · 3.01 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: a b s t r a c t Forty isolates of Listeria monocytogenes were obtained from smoked salmon and production environment in a processing plant. Molecular characterisation evidenced the presence of biotypes corresponding to differences in technological conditions, particularly in salting. The growth behaviour of eight selected strains was evaluated in two media (BHI and Salmon Broth) at different temperatures and salt concentrations. In sub-optimal conditions, the physiological biodiversity of the strains was enhanced, especially in Salmon Broth, underlying the crucial importance of the medium in the evaluation of growth potential.
[Show abstract][Hide abstract] ABSTRACT: To evaluate the effect of oregano essential oil on Listeria monocytogenes cytoplasmic membrane.
Nitroxide free-radical Electron Paramagnetic Resonance was applied on L. monocytogenes after 30 min exposure to oregano essential oil concentrations ranging from 0 to 1.25%. The impact of essential oil on the number of viable cells was evaluated by plate count. Growth dynamics of survivors in BHI and TSB were evaluated by turbidometry. After exposure to essential oil concentrations up to 0.50%, the membrane fluidity was changed and its order increased. When L. monocytogenes was exposed to higher concentrations, membrane order parameters slightly returned to the values of untreated cells. However, when the cells were exposed to EO in the presence of sodium azide, which impairs energy metabolism, the membrane fluidity was progressively enhanced, even at the lowest EO concentration (0.25%). Microbiological analyses confirmed a progressive reduction of viable count, at increasing essential oil concentrations. Both in BHI and TSB, the Lag phase length increased in treated cells with respect to controls, suggesting a cell damage recovery.
The combined approach including microbiological and EPR analyses provided relevant information on membrane modification and cell response to essential oils.
EPR approach was demonstrated to be an effective and helpful tool to comprehend the modifications exerted by essential oil on the bacterial membrane.
Letters in Applied Microbiology 08/2010; 51(2):149-57. · 1.63 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: a b s t r a c t Enterococci, which represent a considerable part of the natural microbiota of Pecorino Abruzzese, were investigated for biochemical activities considered to have a potential role in cheese ripening. Acidifying activity was weak, while interesting differences were identified for proteolytic and peptidolytic capa-bility, particularly on H-Lys-b-NA. The presence of C-4 esterase and C-8 esterase lipase, as well as enzymes such as phosphohydrolases and acid phosphatase was confirmed by means of API ZYM. SPME-GC analyses of volatile compounds revealed the production of ethanol, diacetyl and, of particular interest, acetoin after 15 days at 10 C, with important differences among the species. After 15 days at 10 C, Enterococcus faecalis isolates were generally the most active, giving the highest results for most of the biochemical activities investigated, and also some Enterococcus durans showed interesting metabolic features that might be important to the development of the peculiar sensory properties of this cheese.
International Dairy Journal 01/2010; · 2.33 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The objective of this study was to examine the Saccharomyces and non-Saccharomyces yeast populations involved in a spontaneous fermentation of a traditional high sugar must (Vino cotto) produced in central Italy. Molecular identification of a total of 78 isolates was achieved by a combination of PCR-RFLP of the 5.8S ITS rRNA region and sequencing of the D1/D2 domain of the 26S rRNA gene. In addition, the isolates were differentiated by RAPD-PCR. Only a restricted number of osmotolerant yeast species, i.e. Candida apicola, Candida zemplinina and Zygosaccharomyces bailii, were found throughout all the fermentation process, while Saccharomyces cerevisiae prevailed after 15 days of fermentation. A physiological characterization of isolates was performed in relation to the resistance to osmotic stress and ethanol concentration. The osmotolerant features of C. apicola, C. zemplinina and Z. bailii were confirmed, while S. cerevisiae strains showed three patterns of growth in response to different glucose concentrations (2%, 20%, 40% and 60% w/v). The ability of some C. apicola and C. zemplinina strains to grow at 14% v/v ethanol is noteworthy. The finding that some yeast biotypes with higher multiple stress tolerance can persist in the entire winemaking process suggests possible future candidates as starter for Vino cotto production.
International journal of food microbiology 02/2009; 130(3):179-87. · 3.01 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: High pressure homogenization (HPH) has been proposed as an effective alternative to high hydrostatic pressure in the continuous sanitization of fluid food systems. In this study, we evaluated the influence of HPH treatment, applied individually (one, two or three cycles) or in combination with other mild physical or chemical stresses (mild heat treatment H2O2 and low pH), on the capability of Bacillus cereus and Bacillus subtilis spore, suspended in sterilized double distilled water, to form colonies. Although plate count only slightly decreased in all the strains when one cycle of HPH at 150 MPa was applied alone, the spores released significant levels of dipicolinic acid (up to 28%) that could indicate a possible disruption of spore layers. Three consecutive cycles of HPH determined high reduction of colony count (about 5 log CFU/ml) and high DPA release (52%). Among the stress conditions applied, it was observed that only the thermal shock after one HPH cycle reduced the colony count of 2.3 log CFU/ml and induced a DPA release up to 57%.These results suggested HPH as a novel application for B. cereus and B. subtilis control in fluid foods.
[Show abstract][Hide abstract] ABSTRACT: Some tropicalSaccharomyces cerevisiae strains, isolated fromChampús, a traditional Colombian low alcoholic fermented beverage, were characterised in order to select yeasts for aroma improvement
in wine. H2S production, volatile acidity, β-glucosidase activity, higher alcoholesters and terpenes production were evaluated in this
study. These tropical strains were characterised by a considerable production of ethyl hexanoate, 2-phenylethanol, 2-phenylethyl
acetate, and geraniol, detected by SPME-GC-MS. Odor activity values were calculated to analyse the effects of yeasts strains
on wine aroma, resulting in six distinctive wine groups, as evidenced by discriminant analysis. These results suggest thatSaccharomyces strains isolated fromChampús can be an important source for new tropical yeast biotypes with potential winemaking applications, producing a wide range
of aroma compounds.
Annals of Microbiology 01/2009; 59(4):733-740. · 1.55 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The aim of this study was to identify and characterise the predominant yeasts in Champús, a traditional Colombian cereal-based beverage with a low alcoholic content. Samples of Champús from 20 production sites in the Cauca Valley region were analysed. A total of 235 yeast isolates were identified by conventional microbiological analyses and by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of ITS1-5.8S rDNA-ITS2. The dominant species were: Saccharomyces cerevisiae, Issatchenkia orientalis, Pichia fermentans, Pichia kluyveri var. kluyveri, Zygosaccharomyces fermentati, Torulospora delbruekii, Galactomyces geotrichum and Hanseniaspora spp. Model Champús systems were inoculated with single strains of some isolated sporogenus species and the aromatic profiles were analysed by SPME. Analysis of data showed that Champús strains produced high amounts of esters. The aromatic compounds produced by Saccharomyces and non-Saccharomyces yeasts from Champús can exert a relevant influence on the sensory characteristics of the fermented beverage. The Champús strains could thus represent an important source for new yeast biotypes with potential industrial applications.
[Show abstract][Hide abstract] ABSTRACT: Penicillium brevicompactum, commonly encountered in the indoor air, is known to produce a mycotoxin, mycophenolic acid (MPA). This mould has been isolated from a wide range of foods; considering that we had previously isolated this species from contaminated yoghurt, in this study we have evaluated its growth in yoghurt sweetened with sucrose, fructose and fructose added with fruit pieces. Fungal growth was evaluated monitoring CO(2) production in the headspace during yoghurt storage at 4+/-1, 8+/-1 and 10+/-1 degrees C throughout 21 days. P. brevicompactum grew well in the samples sweetened with fructose at 8 and 10 degrees C. The addition of sucrose influenced the growth negatively, particularly at 4 degrees C. Volatile Organic Compounds (VOC) and MPA production was determined at 8 degrees C in inoculated and uninoculated yoghurt, as well as in liquid malt extract. Differences in VOC profiles and in MPA production were correlated with the age of the fungus and with the growth medium. This study points out for the first time the early qualitative changes in volatile production patterns of a common indoor mould, grown in yoghurt, as well as the production of MPA during storage at refrigeration temperatures.
International Journal of Food Microbiology 08/2008; 127(3):276-83. · 3.43 Impact Factor