Bacteriocins are generally recognized as "natural" compounds able to influence the safety and quality of foods. In the past years, a lot of works have been aimed to the detection, purification and characterisation of bacteriocins, as well as to their use in food preservation strategies. A list of review articles dealing with the application of bacteriocins to the protection of foods of animal origin is also available in literature, but it lacks for a summary on the utilization of bacteriocins in vegetable foods. These biopreservatives can be used in a number of ways in food systems and this paper mainly focuses on the state-of-the-art application of bacteriocins from lactic acid bacteria (LAB) to promote the microbial stability of both fermented and non-fermented vegetable food products using bacteriocinogenic strains as starter cultures, protective cultures or co-cultures and the employment of pure bacteriocins as food additives. In addition, applications of bacteriocins from non-LAB are also reviewed. The scopes of future directions of research are summarised.
Food for Health Ireland, Moorepark Food Research Center, Fermoy, County Cork, Ireland; email: email@example.com.
There is an increased desire for sophisticated foods, whereby consumers harbor higher expectations of health-promoting benefits above basic nutrition. Moreover, there is a move from the adulteration of foods with chemical preservatives toward biopreservation. Such expectations have led scientists to identify novel approaches to satisfy both demands, which utilize bacteriocin and peptide-based solutions. The best known examples of biopreservation involve bacteriocins. However, with the exception of nisin, bacteriocins have received limited use in the food industry. Peptides can be added to foods to improve consumer health. Some of the best known examples are angiotensin I-converting enzyme (ACE)-inhibitory peptides, which inhibit ACE, a key enzyme involved in blood pressure (BP) regulation. To be effective, these peptides must be bioavailable, but by their nature, peptides are degraded by digestion with proteolytic enzymes. This review critically discusses the use and potential of peptides and bacteriocins in food systems in terms of safety, quality, and improvement of human health.
Centro de Referencia para Lactobacilos (CERELA-CONICET), San Miguel de Tucumán, Tucumán, Argentina.
Listeria monocytogenes is a foodborne pathogen causative of opportunistic infections. Listeriosis is associated with severe infections in pregnant women causing abortion or neonatal listeriosis. An alternative to antibiotics are safe novel bacteriocins peptides such as enterocin CRL35 with strong antilisterial activity produced by Enterococcus mundtii CRL35. In the present paper, our goal is to study the effectiveness of this peptide and the producer strain in a murine model of pregnancy-associated listeriosis. A single dose of 5×10(9) colony-forming unit of L. monocytogenes FBUNT (Faculty of Biochemistry-University of Tucumán) resulted in translocation of pathogen to liver and spleen of BALB/c pregnant mice. The maximum level of Listeria was observed on day 3 postinfection. Interestingly, the intragastric administration of enterocin CRL35 significantly reduced the translocation of the pathogen to vital organs. On the other hand, the preadministration of E. mundtii CRL35 slightly inhibited this translocation. Listeria infection caused a significant increase in polymorphonuclear leukocytes at day 3 postinfection compared to the noninfected group. This value was reduced after the administration of enterocin CRL35. No significant changes were observed in either white blood cells or lymphocytes counts. Based on the data presented in the present work enterocin CRL35 would be a promising alternative for the prevention of Listeria infections.
Inhibitory activity of Lactobacillus plantarum LMG P-26358 against Listeria innocua when used as an adjunct starter in the manufacture of cheese.
Susan Mills, L Mariela Serrano, Carmel Griffin, Paula M O'Connor, Gwenda Schaad, Chris Bruining, Colin Hill, R Paul Ross, Wilco C Meijer
Teagasc Food Research, Moorepark, Fermoy, Co, Cork, Ireland.
Lactobacillus plantarum LMG P-26358 isolated from a soft French artisanal cheese produces a potent class IIa bacteriocin with 100% homology to plantaricin 423 and bacteriocidal activity against Listeria innocua and Listeria monocytogenes. The bacteriocin was found to be highly stable at temperatures as high as 100°C and pH ranges from 1-10. While this relatively narrow spectrum bacteriocin also exhibited antimicrobial activity against species of enterococci, it did not inhibit dairy starters including lactococci and lactobacilli when tested by well diffusion assay (WDA). In order to test the suitability of Lb. plantarum LMG P-26358 as an anti-listerial adjunct with nisin-producing lactococci, laboratory-scale cheeses were manufactured. Results indicated that combining Lb. plantarum LMG P-26358 (at 108 colony forming units (cfu)/ml) with a nisin producer is an effective strategy to eliminate the biological indicator strain, L. innocua. Moreover, industrial-scale cheeses also demonstrated that Lb. plantarum LMG P-26358 was much more effective than the nisin producer alone for protection against the indicator. MALDI-TOF mass spectrometry confirmed the presence of plantaricin 423 and nisin in the appropriate cheeses over an 18 week ripening period. A spray-dried fermentate of Lb. plantarum LMG P-26358 also demonstrated potent anti-listerial activity in vitro using L. innocua. Overall, the results suggest that Lb. plantarum LMG P-26358 is a suitable adjunct for use with nisin-producing cultures to improve the safety and quality of dairy products.
Department of Animal Science, The University of Tennessee, Knoxville, TN 37996-4574, USA.
AIMS Several bacteriocins (BCNs) that were identified from chicken commensal bacteria dramatically reduced Campylobacter colonization in poultry and are being directed toward on-farm control of this important foodborne human pathogen. A recent study has shown that BCN resistance in Campylobacter jejuni is very difficult to develop in vitro. In this study, in vivo development and stability of BCN resistance in Campylobacter was examined. METHODS AND RESULTS Chickens infected with Camp. jejuni NCTC 11168 were treated with BCN E-760 at the dose of 5 mg kg(-1) body weight day(-1) via oral gavages for three consecutive days, which selected BCN-resistant (BCN(r)) mutants in the treated birds. However, all the in vivo-selected mutants only displayed low levels of resistance to BCN (MIC = 2-8 mg l(-1)) when compared to parent strain (MIC = 0.5 mg l(-1)). Inactivation of CmeABC efflux pump of the BCN(r) mutants led to increased susceptibility to BCN (8-32 fold MIC reduction). Three different BCN(r) Campylobacter strains (in vitro- or in vivo-derived) were examined for the stability of BCN resistance using both in vitro and in vivo systems. The low level of BCN resistance in these strains was not stable in vitro or in vivo in the absence of BCN selection pressure. CONCLUSIONS Usage of BCN E-760 only selected low-level BCN(r) Camp. jejuni mutants in vivo, and the low-level BCN resistance was not stable in vitro and in vivo. SIGNIFICANCE AND IMPACT OF THE STUDY The study provides helpful information for risk assessment of the future practical application of the anti-Campylobacter BCNs in animals.
SENFIMIZO Department, Section of Phytopathology and Agricultural Microbiology, University of Palermo, Viale delle Scienze 4, 90128 Palermo, Italy.
Six strains of non-starter lactic acid bacteria (NSLAB) were used to extend the shelf-life of the fresh cheese Tosèla manufactured with pasteurised cows' milk. The acidification kinetics of three Lactobacillus paracasei, one Lactobacillus rhamnosus and two Streptococcus macedonicus were studied in synthetic milk medium. Lb. paracasei NdP78 and NdP88 and S. macedonicus NdP1 and PB14-1 showed an interesting acidifying capacity and were further characterised for growth in UHT milk and production of antimicrobial compounds. Lb. paracasei NdP78 and S. macedonicus NdP1 grew more than 2 log cycles in 6 h. Lb. paracasei NdP78 was also found to produce a bacteriocin-like inhibitory substance (BLIS) active against Listeria monocytogenes. The four NSLAB strains (singly or in combination) were used to produce experimental pilot-scale cheeses which were compared by a panel. The cheese manufactured with the mixed culture Lb. paracasei NdP78 - S. macedonicus NdP1 was the most appreciated for its sensory properties. The cheeses produced at factory-scale showed higher concentrations of lactobacilli (7.90 log CFU/g) and streptococci (6.10 log CFU/g), but a lower development of coliforms (3.10 log CFU/g) and staphylococci (2.78 log CFU/g) than control cheese (4.86, 4.89, 4.93 and 5.00 log CFU/g of lactobacilli, streptococci, coliforms and staphylococci, respectively) processed without NSLAB addition. The food pathogens Salmonella spp. and Listeria monocytogenes were never detected. The dominance of the species inoculated was demonstrated by denaturing gradient gel electrophoresis (DGGE), whereas strain recognition was evaluated by randomly amplified polymorphic DNA (RAPD)-PCR. From the results obtained, Lb. paracasei NdP78 and S. macedonicus NdP1 were able to persist during the storage of Tosèla cheese and their combination influenced positively the sensory characteristics and shelf-life of the final product.
Exploration of Islamic medicine plant extracts as powerful antifungals for the prevention of mycotoxigenic Aspergilli growth in organic silage.
Genetic Engineering and Biotechnology Research Institute, Minoufiya University, El-Sadat City, 79/32897, Egypt. firstname.lastname@example.org.
BACKGROUND: Feed contamination with mycotoxins is a major risk factor for animals and humans as several toxins can exist as residues in meat and milk products, giving rise to carry-over to consumers via ingestion of foods of animal origin. The starting point for prevention, in this chain, is to eliminate the growth of mycotoxigenic fungi in the animal forage. Ten plant extracts, recommended in Islamic medicine, were evaluated as antifungal agents against mycotoxigenic Aspergilli, i.e. Aspergillus flavus and A. ochraceus, growth in organic maize silage. RESULTS: Most extracts had remarkable antifungal activities using both qualitative and quantitative evaluation methods. Cress (Lepidium sativum) seed extract was proven to be the most powerful among the plants examined. Blending of the most effective extracts (garden cress seed, pomegranate peel and olive leaf extracts), individually at their minimal fungicidal concentrations, with maize silage resulted in the reduction of inoculated A. flavus colony counts by 99.9, 99.6 and 98.7%, respectively, whereas silage blending with the combined extracts completely prohibited fungal growth for up to 30 days of incubation under aerobic conditions. CONCLUSION: Besides the health promoting effects, silage blending with the bioactive plant extracts examined could lead to the required protection from pathogenic and mycotoxigenic fungi. Copyright © 2011 Society of Chemical Industry.
Bioprocess Biosyst Eng. 2011 Apr 2;: 21461772
Kinetic modelling of the sequential production of lactic acid and xylitol from vine trimming wastes.
Group of Sustainable Processes and Products Engineering and Management, Department of Chemical Engineering/School of Engineering, USC, University of Santiago de Compostela, Rúa Lope Gómez de Marzoa s/n, 15782, Santiago de Compostela, Spain.
A mathematical model describing the kinetics of the sequential production of lactic acid and xylitol from detoxified-concentrated vine trimming hemicellulosic hydrolysates by Lactobacillus rhamnosus and Debaryomyces hansenii, respectively, was developed from the basic principles of mass balance in two stages considering as main reactions:(1) glucose and xylose consumption by L. rhamnosus; and (2) xylitol and arabitol production by D. hansenii. The model allows to evaluate the yields and productivities under microaerobic and oxygen restricted conditions (in particular the effects caused by purging the oxygen with nitrogen), which were particularly important during the xylose to xylitol bioconversion by yeasts. The model was tested using experimental data obtained from detoxified-concentrated hemicellulosic hydrolysates, after CaCO(3) addition in both types of fermentation processes, without purges (microaerobic conditions) or purging oxygen with nitrogen (oxygen-limited conditions) after sampling in order to reduce the oxygen dissolved. L. rhamnosus was removed by microfiltration before adding D. hansenii at the beginning of the second stage. Mass balance-based and logistic functions were successfully applied to develop the model of the system which properly predicts the consumption of sugars as well as the metabolites produced and yields. The dynamics of fermentation were also adequately described by the developed model.
Isolation and characterisation of lactic acid bacteria from yan-jiang (fermented ginger), a traditional fermented food in Taiwan.
Department of Biotechnology, Ming Chuan University, No. 5 De-Ming Road, Gui-Shan, Taoyuan 333, Taiwan.
BACKGROUND Yan-jiang (fermented ginger) is a popular traditional fermented food in Taiwan. The lactic acid bacteria (LAB) microflora in yan-jiang has not been studied in detail. In this study, LAB from yan-jiang were isolated, characterised and identified. RESULTS A total of 176 LAB were isolated; 160 cultures were isolated from yan-jiang samples and 16 cultures were isolated from raw ginger. These isolates were characterised phenotypically and then divided into nine groups (A to I) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Lactobacillus sakei and Lactococcus lactis subsp. lactis were the major LAB found in the initial 2 days of fermentation without pickled plums; these species were mostly replaced by Weissella cibaria and L. plantarum after 3 days of fermentation. In the fermentation bucket with added pickled plums, W. cibaria was the most abundant LAB found during fermentation. The antibacterial activities of the isolates were determined. Twenty-four Lc. lactis subsp. lactis and 19 W. cibaria strains showed inhibitory activity against the indicator strain L. sakei JCM 1157(T). CONCLUSION Results demonstrate that various LAB species were more numerous when fermentation was carried out without pickled plums. LAB also had effects on the aroma and flavour of yan-jiang.
Efficacy of bacteriocin-containing cell-free culture supernatants from lactic acid bacteria to control Listeria monocytogenes in food.
Institut Dr. Erdmann GmbH, Laboratory for food safety and environmental hygiene, Amselweg 12; D-33378 Rheda-Wiedenbrück, Germany. email@example.com
Consumer demands have led to an increased interest in the use of natural antimicrobials for food protection. With the objective of developing novel products for enhancing the microbial safety of food, we have tested cell-free culture supernatants (CFS's) of eight antagonistic bacterial strains for their efficacy to inhibit Listeria monocytogenes in different food matrices. The antagonistic strains represented different members of the order Lactobacillales as well as one isolate of Staphylococcus sciuri and all showed strong inhibition of L. monocytogenes on agar plates. Cell-free supernatants were obtained after growing the bacteria in a yeast extract-glucose broth. In six of the CFS's, different class IIa bacteriocins, namely leucocin A, leucocin B, mundticin L, pediocin PA-1, sakacin A, and sakacin X, were identified as the major anti-listerial compounds. For the other two strains, the active substances could not be ascertained conclusively. The minimal effective concentration (MEC) of the individual CFS's to achieve a 2.3 log(10) reduction of L. monocytogenes was determined in culture broth, whole milk, and ground beef at 4°C. While all bacteriocin-containing CFS's were effective in broth at concentrations from 52 to 205 AU/ml, significant higher concentrations were needed when applied in food. Best results were obtained using CFS's containing pediocin PA-1, that displayed only three- and ten-times higher MEC's in milk (307 AU/ml) and ground meat (1024 AU/g) compared to broth, respectively. A twenty-fold increase in the MEC (2048 AU/ml) was observed for a mundticin L-containing fermentate, and a CFS containing leucocin A and B was inactivated more than fifty-fold (>1280 AU/ml) in both food matrices. Remarkably, the sakacin A and sakacin X containing CFS's displayed very selective inactivation rates, in which sakacin A was only effective in meat (512 AU/g), while sakacin X was only effective in milk (2048 AU/ml). In all cases, inhibition of L. monocytogenes was only transient and surviving or resistant bacteria started growing after prolonged storage. These results highlight the importance of careful testing the effectiveness of bacteriocins in the food systems for which they are intended to be applied against the selected target and non-target bacteria. Furthermore, the outgrowth of surviving or resistant bacterial populations points out that the tested bacteriocins are not suited to assure full inhibition of L. monocytogenes in a food product, if not applied in combination with additional preservative measures.
LAB-Secretome: a genome-scale comparative analysis of the predicted extracellular and surface-associated proteins of Lactic Acid Bacteria.
Centre for Molecular and Biomolecular Informatics, Radboud University Nijmegen Medical Centre, PO Box 9101, 6500 HB Nijmegen, The Netherlands. firstname.lastname@example.org
BACKGROUND In Lactic Acid Bacteria (LAB), the extracellular and surface-associated proteins can be involved in processes such as cell wall metabolism, degradation and uptake of nutrients, communication and binding to substrates or hosts. A genome-scale comparative study of these proteins (secretomes) can provide vast information towards the understanding of the molecular evolution, diversity, function and adaptation of LAB to their specific environmental niches. RESULTS We have performed an extensive prediction and comparison of the secretomes from 26 sequenced LAB genomes. A new approach to detect homolog clusters of secretome proteins (LaCOGs) was designed by integrating protein subcellular location prediction and homology clustering methods. The initial clusters were further adjusted semi-manually based on multiple sequence alignments, domain compositions, pseudogene analysis and biological function of the proteins. Ubiquitous protein families were identified, as well as species-specific, strain-specific, and niche-specific LaCOGs. Comparative analysis of protein subfamilies has shown that the distribution and functional specificity of LaCOGs could be used to explain many niche-specific phenotypes.A comprehensive and user-friendly database LAB-Secretome was constructed to store, visualize and update the extracellular proteins and LaCOGs http://www.cmbi.ru.nl/lab_secretome/. This database will be updated regularly when new bacterial genomes become available. CONCLUSIONS The LAB-Secretome database could be used to understand the evolution and adaptation of lactic acid bacteria to their environmental niches, to improve protein functional annotation and to serve as basis for targeted experimental studies.
Other papers by authors:
Yeast microbiota associated with spontaneous sourdough fermentations in the production of traditional wheat sourdough breads of the Abruzzo region (Italy).
Department of Food Science, University of Teramo, Via CR Lerici 1, 64023 Mosciano Sant'Angelo, Teramo, Italy.
The aims of this study were to describe the yeast community of 20 sourdoughs collected from central Italy and to characterize the sourdoughs based on chemical properties. A polyphasic approach consisting of traditional culture-based tests (spore-forming and physiological tests) and molecular techniques (PCR-RFLP, RAPD-PCR, PCR-DGGE) and chemical analysis (total acidity, acids, and sugar contents), was utilized to describe the yeast population and to investigate the chemical composition of the doughs. PCR-RFLP analysis identified 85% of the isolates as Saccharomyces cerevisiae, with the other dominant species being Candida milleri (11%), Candida krusei (2.5%), and Torulaspora delbrueckii (1%). RAPD-PCR analysis, performed with primers M13 and LA1, highlighted intraspecific polymorphism among the S. cerevisiae strains. The diversity of the sourdoughs from the Abruzzo region is reflected in the chemical composition, yeast species, and strain polymorphism. Our approach using a combination of phenotypic and genotypic methods identified the yeast species in the 20 sourdough samples and provided a complete overview of the yeast populations found in sourdoughs from the Abruzzo region.
The role of environmental factors and medium composition on bacteriocin-like inhibitory substances (BLIS) production by Enterococcus mundtii strains.
Dipartimento di Scienze degli Alimenti, Sezione di Microbiologia Agro-Alimentare ed Ambientale, Università degli Studi di Teramo, V. C.R. Lerici 1, 64023 Mosciano Sant'Angelo (TE), Italy.
Bacteriocin-like inhibitory substances (BLIS)-producers Enterococcus mundtii WGWT1-1A, WGW11.2, WGJ20.1, WGJ40.2 and WGK53 from raw material origin were subjected to a study for the characterization of antimicrobial compound production under several growth conditions, including different cultivation media, growth temperatures, pHs, different concentrations and sources of nitrogen compounds, carbohydrates and other nutritional factors, and in the presence of different percentages of ethanol and NaCl. The five E. mundtii strains showed different behaviors. However, in all cases, MRS and sour dough bacteria (SDB) were found as the optimal media for BLIS production. In general, the higher BLIS production was observed with pH in the range 6.0-8.0 and, except 45 degrees C, the temperature did not show a defining effect. Low or no BLIS activity was detected after growth without nitrogen sources and carbohydrates. Absence of Tween 80, triammoniun citrate, K2HPO4, MgSO4 and MnSO4 did not affect BLIS activity levels. Except for a strain (WGWT1-1A), ethanol did not play a negative role in BLIS expression, while NaCl determined decrease of BLIS activity, proportional with concentration. The above strains did not contain plasmids, hence, BLIS expression is encoded by chromosomal DNA.
A taxonomic survey of lactic acid bacteria isolated from wheat (Triticum durum) kernels and non-conventional flours.
Aldo Corsetti, Luca Settanni, Clemencia Chaves López, Giovanna E Felis, Mario Mastrangelo, Giovanna Suzzi
In order to explore the correspondence between raw material- and mature sourdough-lactic acid bacterial (LAB) communities, 59 Italian wheat (Triticum durum) grain samples, one bran and six non-conventional flour samples were analyzed through a culture-dependent approach. The highest cell count by an agar medium specific for LAB was 2.16logCFU/g. From about 2300 presumptive LAB (Gram-positive and catalase-negative) colonies collected, a total of 356 isolates were subjected to identification by a genetic polyphasic strategy consisting of RAPD-PCR analysis, partial 16S rRNA gene sequencing, species-specific and multiplex PCRs. The isolates were recognized as 137 strains belonging to Aerococcus, Enterococcus, Lactobacillus, Lactococcus and Pediococcus genera and a phylogram based on partial 16S rRNA genes was constructed. The species most frequently found were Enterococcus faecium, Enterococcus mundtii and Lactobacillus graminis, which are not generally reported to be typical in mature sourdoughs.
Identification of subdominant sourdough lactic acid bacteria and their evolution during laboratory-scale fermentations.
Dipartimento di Scienze degli Alimenti, Sezione di Microbiologia Agro-Alimentare ed Ambientale, Università degli Studi di Teramo, V.C.R. Lerici 1, 64023 Mosciano Sant’Angelo, Teramo, Italy.
Presumptive lactic acid bacterial cocci were found in six sourdoughs (out of 20) from the Abruzzo region (central Italy) and subjected to phenotypic and genotypic characterization. A total of 21 isolates, recognized as seven strains by randomly amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) typing, were identified by a polyphasic approach, consisting of 16S rRNA gene sequencing, multiplex PCR assays and physiological features, as Enterococcus faecium and Pediococcus pentosaceus. Four strains belonging to those species and previously isolated from wheat kernels were inoculated in sterile flour to verify their capacity to grow in sourdough environment. Doughs with several dual bacterial combinations, including Lactobacillus sanfranciscensis, were propagated for 11 days and pH measurements and bacterial counts were carried out.
The Use of Lactobacillus pentosus 1MO To Shorten the Debittering Process Time of Black Table Olives (Cv. Itrana and Leccino): A Pilot-Scale Application.
Maurizio Servili, Luca Settanni, Gianluca Veneziani, Sonia Esposto, Ombretta Massitti, Agnese Taticchi, Stefania Urbani, Gian Montedoro, Aldo Corsetti
Dipartimento di Scienze Economico-Estimative e degli Alimenti, Sezione di Tecnologie e Biotecnologie degli Alimenti, Università degli Studi di Perugia, Perugia, Italy, and Dipartimento di Scienze degli Alimenti, Sezione di Microbiologia Agro-Alimentare ed Ambientale, Università degli Studi di Teramo, Teramo, Italy.
Fifty lactobacilli isolated from black table olive brines were evaluated for their salt tolerance, resistance to oleuropein and verbascoside, and ability to grow in modified filter-sterilized brines. A strain of Lactobacillus pentosus was selected and used as a starter to ferment, in pilot plant, black olives (Itrana and Leccino cv.) in brines modified for pH, carbohydrate, and growth factor concentrations, at 28 degrees C. The temperature-controlled fermentation of Leccino cv. olives resulted in obtaining ready-to-eat, high-quality table olives in a reduced-time process. HPLC analysis of phenolic compounds from fermented olives showed a decrease of oleuropein, a glucoside secoiridoid responsible for the bitter taste of olive drupes, and an increase of the hydroxytyrosol concentration. The selected strain of L. pentosus (1MO) allowed the reduction of the debittering phase period to 8 days.
Combination of Multiplex PCR and PCR-Denaturing Gradient Gel Electrophoresis for Monitoring Common Sourdough-Associated Lactobacillus Species.
Luca Settanni, Sara Valmorri, Douwe van Sinderen, Giovanna Suzzi, Antonello Paparella, Aldo Corsetti
Dipartimento di Scienze degli Alimenti, Sezione di Microbiologia Agro-Alimentare ed Ambientale, Università degli Studi di Teramo, V. C. R. Lerici 1, 64023 Mosciano Sant'Angelo, Teramo, Italy. email@example.com.
A combination of denaturing gradient gel electrophoresis (DGGE) and a previously described multiplex PCR approach was employed to detect sourdough lactobacilli. Primers specific for certain groups of Lactobacillus spp. were used to amplify fragments, which were analyzed by DGGE. DGGE profiles obtained from Lactobacillus type strains acted as standards to analyze lactobacilli from four regional Abruzzo (central Italy) sourdoughs.
Dipartimento di Scienze degli Alimenti, Sezione di Tecnologie e Biotecnologie degli Alimenti, Università degli Studi di Perugia, Perugia, Italy
A two-step multiplex PCR-based method was designed for the rapid detection of 16 species of lactobacilli known to be commonly present in sourdough. The first step of multiplex PCR was developed with a mixture of group-specific primers, while the second step included three multiplex PCR assays with a mixture of species-specific primers. Primers were derived from sequences that specify the 16S rRNA, the 16S-23S rRNA intergenic spacer region, and part of the 23S rRNA gene. The primer pairs designed were shown to exclusively amplify the targeted rrn operon fragment of the corresponding species. Due to the reliability of simultaneously identifying Lactobacillus plantarum, Lactobacillus pentosus, and Lactobacillus paraplantarum, a previously described multiplex PCR method employing recA gene-derived primers was included in the multiplex PCR system. The combination of a newly developed, quick bacterial DNA extraction method from sourdough and this multiplex PCR assay allows the rapid in situ detection of several sourdough-associated lactobacilli, including the recently described species Lactobacillus rossii, and thus represents a very useful alternative to culture-based methodologies.
Aldo Corsetti, Luca Settanni, Douwe van Sinderen, Giovanna E Felis, Franco Dellaglio, Marco Gobbetti
Dipartimento di Scienze degli Alimenti, Sezione di Tecnologie e Biotecnologie degli Alimenti, Università degli Studi di Perugia, Perugia, Italy.
Screening of sourdough lactic acid bacteria for bacteriocin production resulted in the isolation of a Gram-positive, catalase-negative, non-spore-forming, non-motile rod bacterium (strain CS1T) that could not be associated with any previously described species. Comparative 16S rRNA gene sequence analysis recognized strain CS1T as a distinct member of the genus Lactobacillus. By a species-specific PCR strategy, five additional strains previously isolated from sourdoughs were found to belong to the same species as strain CS1T, as confirmed by 16S rRNA gene sequence analysis. The closest related species were Lactobacillus durianis, Lactobacillus malefermentans and Lactobacillus suebicus, with which strain CS1T shared 93 % sequence similarity. For a further characterization of strain CS1T, physiological (growth temperature, CO2 production, hydrolysis of arginine, isomeric type of lactate, sugar fermentation) and chemotaxonomic (G+C content and peptidoglycan structure) properties were determined. Phenotypic characterization showed that strain CS1T was a member of the obligately heterofermentative group of the genus Lactobacillus. The DNA G+C content was 44.6 mol%. The peptidoglycan was of the A3alpha (L-Lys-L-Ser-L-Ala2) type. Physiological, biochemical and genotypic data, as well as results of DNA-DNA hybridization of genomic DNA with one of the closest phylogenetic relatives, L. durianis (34.3 %), indicated that strain CS1T represents a novel species of the genus Lactobacillus for which the name Lactobacillus rossii sp. nov. is proposed. The type strain of this species is CS1T (=ATCC BAA-822T=DSM 15814T).
J Food Sci. 2012 Aug 21;: 22908955
Authors are with Dept. of Scienze degli Alimenti, Univ. degli Studi di Teramo, Via C.R. Lerici 1, 64023 Mosciano Sant'Angelo (Teramo), Italy. Direct inquiries to author Tofalo (E-mail: firstname.lastname@example.org).
The question if the "Brett character" is a favorable wine attribute is one of the most controversial issues and it is currently addressed by many researches. Actually, the presence of Brettanomyces/Dekkera in wine during barrel aging is often associated to detrimental organoleptic characteristics depending on the release of volatile phenols (for example, 4-ethylphenol and 4-ethylguaiacol); for that reason the possibility to rapidly detect the yeast at the early stage of wine production could allow preventive actions to reduce wine spoilage. In this work, 25 and 5 samples from conventional and organic vineyards, respectively, all suspected to be spoiled by Brettanomyces/Dekkera spp., were analyzed using both culture-dependent and culture-independent techniques. In particular, a DNA extraction protocol and a real-time quantitative PCR (qPCR) assay to directly detect and quantify B. bruxellensis were optimized. Results showed that B. bruxellensis was present in 22 of 30 samples, ranging from 10 to 10(4) CFU/mL, lower values being found in organic wines (10 to 10(2) CFU/mL). Overall, qPCR was proved to be a useful and valuable wine control system, since 12 samples were recorded as positive for yeast presence when analyzed by qPCR and negative in case of plate count analyses. Practical Application: Brettanomyces cells were detected using a qPCR method, optimized in this study, which allows to obtain results quickly.
Front Microbiol. 2012 ;3 :248 22833739
Application of starter cultures to table olive fermentation: an overview on the experimental studies.
Department of Food Science, University of Teramo, Mosciano Sant'Angelo, Teramo, Italy.
Table olives are one of the oldest fermented foods and are considered as an important component of the Mediterranean diet, since their richness in monounsaturated fats (primarily oleic acid) and phenolic compounds may function as antioxidants in the human body; in the Western world they represent one of the most popular fermented vegetables but, despite its economic significance, table olive fermentation is still craft-based and empirical. In particular, such a type of fermentation results from the competitive activities among indigenous, contaminating microorganisms, the microbial balance depending on several intrinsic (pH, water activity, diffusion of nutrients from the drupe, and level of anti-microbial compounds) and extrinsic (temperature, oxygen availability, and salt concentration) factors. At present, to reduce the risk of spoilage and to achieve a more predictable process there is an increasing interest in developing starter cultures for table olives fermentation. Anyway, the application of starter cultures in the field of table olives is quite far from reaching the diffusion as it has in other sectors of food industry (e.g., dairy products and alcoholic beverages). This review focuses on experimental researches devoted to studying starter cultures for possible application to table olive fermentation both at artisan and industrial level.
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Department of Analytical Chemistry, Nutrition and Food Science, School of Veterinary Sciences, University of Santiago de Compostela, E-27002 Lugo, Spain. email@example.com.
Most types of bacteria produce bacteriocins, which are proteinaceous extracellular compounds that can inhibit the growth of other undesirable microorganisms. Bacteriocins are receiving increasing attention, due to their many applications, ranging from their initial application in strategies for food preservation to more recent proposed uses in biomedical strategies aimed at fighting certain bacterial infections. Thus, while nisin has a long history of use as a safe additive in certain food products for the purpose of food preservation, certain bacteriocin-producing lactic acid bacteria, which are generally recognised as safe microorganisms, or their extracellular extracts are receiving increased attention as protective cultures or antimicrobial extracts in minimally processed food products. More recently, a number of these bacteriocin-producing cultures have been proposed for use in other applications, such as in probiotics, for the inhibition of biofilms in the food industry, or even as coadjuvants of combined therapeutical strategies along with other antimicrobial agents in biomedical applications. This review aims to provide a brief overview of the most relevant recent patents in this field.
Plant extracts for the control of bacterial growth: efficacy, stability and safety issues for food application.
Human Resource Development Department, Council of Scientific and Industrial Research-Central Food Technological Research Institute, Mysore 570020, India. firstname.lastname@example.org
The microbial safety of foods continues to be a major concern to consumers, regulatory agencies and food industries throughout the world. Many food preservation strategies have been used traditionally for the control of microbial spoilage in foods but the contamination of food and spoilage by microorganisms is a problem yet to be controlled adequately. Although synthetic antimicrobials are approved in many countries, the recent trend has been for use of natural preservatives, which necessitates the exploration of alternative sources of safe, effective and acceptable natural preservatives. Plants contain innumerable constituents and are valuable sources of new and biologically active molecules possessing antimicrobial properties. Plants extracts either as standardized extracts or as a source of pure compounds provide unlimited opportunities for control of microbial growth owing to their chemical diversity. Many plant extracts possess antimicrobial activity against a range of bacteria, yeast and molds, but the variations in quality and quantity of their bioactive constituents is the major detriments in their food use. Further, phytochemicals added to foods may be lost by various processing techniques. Several plant extracts or purified compounds intended for food use have been consumed by humans for thousands of years, but typical toxicological information is not available for them. Although international guidelines exist for the safety evaluation of food additives, owing to problems in standardization of plant extracts, typical toxicological values have not been assigned to them. Development of cost effective isolation procedures that yield standardized extracts as well as safety and toxicology evaluation of these antimicrobials requires a deeper investigation.
Technische Universität Berlin, Fachgebiet für Angewandte und Molekulare Mikrobiologie, Germany. email@example.com
In this review article, the beneficial application of bacterial spore formers as probiotics in the food industry is discussed based on the knowledge gleaned from current publications. The summary of new scientific results provides evidence of the advantages of the utilisation of Bacillus or Clostridium strains in the food industry. Both bacteria are able to produce a very stable duration form: the endospore. Compared to the widely used lactic acid bacteria, bacterial spores offer the advantage of a higher survival rate during the acidic stomach passage and better stability during the processing and storage of the food product. In many food products, germination of the spores does not occur. Hence the product quality of the food is not affected because of their inactive metabolism. Besides the possible utilisation and functional properties, an overview of the fast-developing knowledge about the mechanisms of the beneficial health effects of spore-forming bacteria is provided.
François Bourdichon, Serge Casaregola, Choreh Farrokh, Jens C Frisvad, Monica L Gerds, Walter P Hammes, James Harnett, Geert Huys, Svend Laulund, Arthur Ouwehand, Ian B Powell, Jashbhai B Prajapati, Yasuyuki Seto, Eelko Ter Schure, Aart Van Boven, Vanessa Vankerckhoven, Annabelle Zgoda, Sandra Tuijtelaars, Egon Bech Hansen
Danone Research, RD128, 91 767 Palaiseau Cedex, France.
Microbial food cultures have directly or indirectly come under various regulatory frameworks in the course of the last decades. Several of those regulatory frameworks put emphasis on "the history of use","traditional food", or "general recognition of safety". Authoritative lists of microorganisms with a documented use in food have therefore come into high demand. One such list was published in 2002 as a result of a joint project between the International Dairy Federation (IDF) and the European Food and Feed Cultures Association (EFFCA). The "2002 IDF inventory" has become a de facto reference for food cultures in practical use. However, as the focus mainly was on commercially available dairy cultures, there was an unmet need for a list with a wider scope. We present an updated inventory of microorganisms used in food fermentations covering a wide range of food matrices (dairy, meat, fish, vegetables, legumes, cereals, beverages, and vinegar). We have also reviewed and updated the taxonomy of the microorganisms used in food fermentations in order to bring the taxonomy in agreement with the current standing in nomenclature.
J G LeBlanc, J E Laiño, M Juarez del Valle, V Vannini, D van Sinderen, M P Taranto, G Font de Valdez, G Savoy de Giori, F Sesma
Centro de Referencia para Lactobacilos (CERELA-CONICET), Chacabuco 145, Tucumán, Argentina. firstname.lastname@example.org
Although most vitamins are present in a variety of foods, human vitamin deficiencies still occur in many countries, mainly because of malnutrition not only as a result of insufficient food intake but also because of unbalanced diets. Even though most lactic acid bacteria (LAB) are auxotrophic for several vitamins, it is now known that certain strains have the capability to synthesize water-soluble vitamins such as those included in the B-group (folates, riboflavin and vitamin B(12) amongst others). This review article will show the current knowledge of vitamin biosynthesis by LAB and show how the proper selection of starter cultures and probiotic strains could be useful in preventing clinical and subclinical vitamin deficiencies. Here, several examples will be presented where vitamin-producing LAB led to the elaboration of novel fermented foods with increased and bioavailable vitamins. In addition, the use of genetic engineering strategies to increase vitamin production or to create novel vitamin-producing strains will also be discussed. This review will show that the use of vitamin-producing LAB could be a cost-effective alternative to current vitamin fortification programmes and be useful in the elaboration of novel vitamin-enriched products.
Ming Li, Dorothy L Patton, Yvonne Cosgrove-Sweeney, Deena Ratner, Lisa C Rohan, Alexander M Cole, Patrick M Tarwater, Phalguni Gupta, Bharat Ramratnam
Laboratory of Retrovirology, Division of Infectious Diseases, Department of Medicine, Rhode Island and Miriam Hospitals, Warren Alpert Medical School of Brown University, Providence, RI, USA.
An urgent need exists for HIV-1 microbicides. Here, we describe the in vivo testing of lactic acid bacteria bioengineered to secrete cyanovirin-N. We fed pigtail macaques a yogurt formulation that used bioengineered strains as a starter culture. Cyanovirin-N expression could be detected in the rectal vault during and immediately after feeding. Ex vivo viral challenge of rectal tissue biopsies revealed that peak viral burden was significantly lower in tissue obtained from experimental animals compared with control animals. Formulation of candidate compounds in lactic acid bacteria and their oral administration seems to be a feasible strategy for mucosal delivery of microbicides.
J Food Sci. ;75 (9):E596-604 21535594
Suwimon Keeratipibul, Naphatrapi Luangsakul, Shinya Otsuka, Shigeru Sakai, Yasushi Hatano, Somboon Tanasupawat
Dept. of Food Technology, Faculty of Science, Chulalongkorn Univ., Bangkok 10330, Thailand. Suwimon.K@chula.ac.th
The application of Chinese steamed bun starter dough (CSB-SD) in breadmaking was investigated. The activation of CSB-SD to activate the growth of lactic acid bacteria (LAB) and to increase the number of yeast, prior to making bread, was conducted by mixing CSB-SD with wheat flour and water and then incubating for 24 h. Wheat flour was then substituted by this activated CSB-SD (aCSB-SD) at 10%, 30%, and 50%(w/w) to make bread. Dough and bread properties were studied comparing to the control (without aCSB-SD). From the farinograph results, a high aCSB-SD substitution level resulted in a less stability in dough with a higher degree of softening. Extensigraph results suggested that after aging, all the substituted dough yielded a greater resistance to extension with lower extensibility values than the control. Substitutions with 30% and 50%(w/w) aCSB-SD significantly increased the total CO(2) gas generation. Scanning electron microscopy SEM images of the 30% and 50%(w/w) substituted dough showed a well-developed gluten matrix. The 50%(w/w) substituted breads obtained a greater risen volume, finer crumb grain, and retained more softness after 5-d storage than the control. In addition, both the 30% and 50%(w/w) substituted breads showed a slightly increased mold stability, as compared to the 0% and 10%(w/w) substituted breads.
Anaerobe. 2011 Apr 8;: 21497663
Microbial ecology and quality assurance in food fermentation systems. The case of kefir grains application.
Democritus University of Thrace, Faculty of Agriculture Development, Laboratory of Microbiology, Biotechnology and Hygiene, 193 Pantazidou str. GR-68200 Orestiada, Greece.
Fermentation technology has become a modern method for food production the last decades as a process for enhancing product stability, safety and sensory standards. The main reason for this development is the increasing consumers' demand for safe and high quality food products. The above has led the scientific community to the thorough study for the appropriate selection of specific microorganisms with desirable properties such as bacteriocin production, and probiotic properties. The main food products produced through fermentation activity are bread, wine, beer cheese and other dairy products. The microorganisms conducting the above processes are mainly yeasts and lactic acid bacteria. The end products of carbohydrate catabolism by these microorganisms contribute not only to preservation as it was believed years ago, but also to the flavour, aroma and texture and to the increase of the nutritional quality by thereby helping determine unique product characteristics. Thus, controlling the function of specific microorganisms or the succession of microorganisms that dominate the microflora is therefore advantageous, because it can increase product quality, functionality and value. Throughout the process of the discovery of microbiological diversity in various fermented food systems, the development of starter culture technology has gained more scientific attention, and it could be used for the control of the manufacturing operation, and management of product quality. In the frame of this review the presentation of the quality enhancement of most consumed fermented food products around the world is attempted and the new trends in production of fermented food products, such as bread is discussed. The review is focused in kefir grains application in bread production.
Departamento de Ciencias de la Salud, Área de Microbiología, Facultad de Ciencias Experimentales, Universidad de Jaén, Jaén, Spain.
Members of the genus Bacillus are known to produce a wide arsenal of antimicrobial substances, including peptide and lipopeptide antibiotics, and bacteriocins. Many of the Bacillus bacteriocins belong to the lantibiotics, a category of post-translationally modified peptides widely disseminated among different bacterial clades. Lantibiotics are among the best-characterized antimicrobial peptides at the levels of peptide structure, genetic determinants and biosynthesis mechanisms. Members of the genus Bacillus also produce many other nonmodified bacteriocins, some of which resemble the pediocin-like bacteriocins of the lactic acid bacteria (LAB), while others show completely novel peptide sequences. Bacillus bacteriocins are increasingly becoming more important due to their sometimes broader spectra of inhibition (as compared with most LAB bacteriocins), which may include Gram-negative bacteria, yeasts or fungi, in addition to Gram-positive species, some of which are known to be pathogenic to humans and/or animals. The present review provides a general overview of Bacillus bacteriocins, including primary structure, biochemical and genetic characterization, classification and potential applications in food preservation as natural preservatives and in human and animal health as alternatives to conventional antibiotics. Furthermore, it addresses their environmental applications, such as bioprotection against the pre- and post-harvest decay of vegetables, or as plant growth promoters.
SENFIMIZO Department, Section of Phytopathology and Agricultural Microbiology, University of Palermo, Viale delle Scienze 4, 90128 Palermo, Italy. email@example.com
Non-starter lactic acid bacteria (NSLAB) dominate cheese microbiota during ripening. They tolerate the hostile environment well and strongly influence the biochemistry of curd maturation, contributing to the development of the final characteristics of cheese. Several NSLAB are selected on the basis of their health benefits (enhancement of intestinal probiosis, production of bioactive peptides, generation of gamma-aminobutyric acid and inactivation of antigenotoxins) and are employed in cheese-making. This review describes the ecology of NSLAB, and focuses on their application as adjunct cultures, in order to drive the ripening process and promote health advantages. The scopes of future directions of research are summarised.