Person: Cintas Izarra, Luis Miguel
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First Name
Luis Miguel
Last Name
Cintas Izarra
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Veterinaria
Department
Nutrición y Ciencia de los Alimentos
Area
Nutrición y Bromatología
Identifiers
16 results
Search Results
Now showing 1 - 10 of 16
Item Draft Genome Sequence of Lactococcus lactis Subsp. cremoris WA2-67: A Promising Nisin-Producing Probiotic Strain Isolated from the Rearing Environment of a Spanish Rainbow Trout (Oncorhynchus mykiss, Walbaum) Farm(Microorganisms, 2022) Feito Hermida, Javier; Contente, Diogo; Ponce Alonso, Manuel; Díaz Formoso, Lara; Araújo, Carlos; Peña Vidal, Nuria; Borrero Del Pino, Juan; Gómez Sala, Beatriz; Campo, Rosa del; Muñoz Atienza, Estefanía; Hernández Cruza, Pablo Elpidio; Cintas Izarra, Luis MiguelProbiotics are a viable alternative to traditional chemotherapy agents to control infectious diseases in aquaculture. In this regard, Lactococcus lactis subsp. cremoris WA2-67 has previously demonstrated several probiotic features, such as a strong antimicrobial activity against ichthyopathogens, survival in freshwater, resistance to fish bile and low pH, and hydrophobicity. The aim of this manuscript is an in silico analysis of the whole-genome sequence (WGS) of this strain to gain deeper insights into its probiotic properties and their genetic basis. Genomic DNA was purified, and libraries prepared for Illumina sequencing. After trimming and assembly, resulting contigs were subjected to bioinformatic analyses. The draft genome of L. cremoris WA2-67 consists of 30 contigs (2,573,139 bp), and a total number of 2493 coding DNA sequences (CDSs). Via in silico analysis, the bacteriocinogenic genetic clusters encoding the lantibiotic nisin Z (NisZ) and two new bacteriocins were identified, in addition to several probiotic traits, such as the production of vitamins, amino acids, adhesion/aggregation, and stress resistance factors, as well as the absence of transferable antibiotic resistance determinants and genes encoding detrimental enzymatic activities and virulence factors. These results unveil diverse beneficial properties that support the use of L. cremoris WA2-67 as a probiotic for aquaculture.Item Antimicrobial activity and occurrence of bacteriocin structural genes in Enterococcus spp. of human and animal origin isolated in Portugal(Archives of Microbiology, 2010) Brandão, Andreia; Almeida, Tereza; Muñoz Atienza, Estefanía; Torres, Carmen; Igrejas, Gilberto; Hernández Cruza, Pablo Elpidio; Cintas Izarra, Luis Miguel; Poeta, Patricia; Herranz Sorribes, CarmenThe main objective of this study was to detect the antimicrobial activity and the presence of bacteriocin structural genes in 224 enterococcal isolates from fecal origin obtained from humans, pets, wild animals and birds. Direct antimicrobial activity against Listeria monocytogenes CECT4032 was detected in 102 (45.6%) of the tested isolates. From these, only 22 displayed bacteriocin activity against this indicator. The bacteriocinogenic strains contained one or more of the bacteriocin structural genes tested in this study, with those of enterocins P, A and L50 (L50A and L50B) being the most abundant. Our results show a high occurrence of the combination of different bacteriocin structural genes in the enterococcal isolates analyzed, indicating an elevated genetic potential of these strains to produce various bacteriocins.Item In vitro and in vivo evaluation of lactic acid bacteria of aquatic origin as probiotics for turbot (Scophthalmus maximus L.) farming(Fish & Shellfish Immunology, 2014) Muñoz Atienza, Estefanía; Araújo, Carlos; Magadán, Susana; Hernández Cruza, Pablo Elpidio; Herranz Sorribes, Carmen; Santos, Ysabel; Cintas Izarra, Luis MiguelTurbot (Scophthalmus maximus L.) is an important commercial marine flatfish. Its production may be affected by bacterial diseases that cause severe economical losses, mainly tenacibaculosis and vibriosis, provoked by Tenacibaculum maritimum and Vibrio splendidus, respectively. An alternative or complementary strategy to chemotherapy and vaccination for the control of these diseases is the use of probiotics. In this work, we report the in vitro and in vivo potential of eight lactic acid bacteria (LAB), previously isolated from fish, seafood and fish products intended for human consumption, as turbot probiotics. Seven out of the eight LAB exerted direct antimicrobial activity against, at least, four strains of T. maritimum and V. splendidus. All LAB survived in seawater at 18 °C for 7 days, and withstood exposure to pH 3.0 and 10% (v/v) turbot bile; however, they differed in cell surface hydrophobicity (8.2–21.7%) and in their ability to adhere to turbot skin (1.2–21.7%) and intestinal (0.7–2.1%) mucus. Most of the tested strains inhibited the binding of turbot pathogens to the mucus. Leuconostoc mesenteroides subsp. cremoris SMM69 and Weissella cibaria P71 were selected based on their strong antimicrobial activity against T. maritimum and V. splendidus, good probiotic properties, and different adhesion ability to skin mucus and capacity to inhibit the adhesion of turbot pathogens to mucus. These two LAB strains were harmless when administered by bath to turbot larvae and juveniles; moreover, real-time PCR on the transcription levels of the immunity-related genes encoding IL-1β, TNF-α, lysozyme, C3, MHC-Iα and MHC-IIα in five organs (head-kidney, spleen, liver, intestine and skin) revealed the ability of these LAB to stimulate their expression in turbot juveniles, especially the non-specific immunity associated genes in mucosal tissues. Based on our results, Lc. cremoris SMM69 and W. cibaria P71 may be considered as suitable probiotic candidates for turbot farming.Item Phenotypic and genetic evaluations of biogenic amine production by lactic acid bacteria isolated from fish and fish products(International Journal of Food Microbiology, 2011) Muñoz Atienza, Estefanía; Landeta, G.; de las Rivas, B.; Gómez Sala, Beatriz; Muñoz, R.; Hernández Cruza, Pablo Elpidio; Cintas Izarra, Luis Miguel; Herranz Sorribes, CarmenIn this work, biogenic amine production (histamine, tyramine and putrescine) by a collection of 74 lactic acid bacteria of aquatic origin has been investigated by means of amino acid decarboxylation by growth on decarboxylase differential medium, biogenic amine detection by thin-layer chromatography (TLC) and decarboxylase gene detection by PCR. None of the evaluated strains showed neither production of histamine and putrescine, nor presence of the genetic determinants encoding the corresponding decarboxylase activities. However, the tyrosine decarboxylase gene (tdc) was present in all the enterococcal strains, and tyramine production was detected by TLC in all of them but Enterococcus faecium BCS59 and MV5. Analysis of the tyrosine decarboxylase operon of these strains revealed the presence of an insertion sequence upstream tdc that could be responsible for their lack of tyrosine decarboxylase activity.Item Safety assessment and molecular genetic profiling by pulsed-field gel electrophoresis (PFGE) and PCR-based techniques of Enterococcus faecium strains of food origin(LWT - Food Science and Technology, 2015) Muñoz Atienza, Estefanía; Dias Araujo, C.; Campo, R.D.; Hernández Cruza, Pablo Elpidio; Herranz Sorribes, Carmen; Cintas Izarra, Luis MiguelEnterococcus faecium is authorized as animal probiotic in the European Union, but this species has emerged as an important cause of nosocomial infections in humans. We investigated the safety of 14 potential probiotic E. faecium strains with antimicrobial activity, previously isolated from food, following the guidance proposed by EFSA. All the enterococci were susceptible to ampicillin, and none of them harbored the genes encoding the enterococcal surface protein (esp), putative glycosyl hydrolase (hylEfm), and insertion sequence IS16. The genetic relatedness of these enterococci was determined by pulsed-field gel electrophoresis (PFGE), random amplified polymorphic DNA (RAPD), enterobacterial repetitive intergenic consensus (ERIC-PCR), and restriction analysis of amplified 16S rDNA (ARDRA). PFGE analysis of SmaI patterns evidenced four subgroups, whereas RAPD and ERIC-PCR analysis gave nine and eight different subgroups, respectively. ERIC-PCR yielded the highest diversity, followed by RAPD and PFGE, while ARDRA achieved the lowest diversity. In conclusion, we demonstrated the absence of well-known enterococcal virulence markers in a collection of E. faecium strains from food, which renders them safe to be used in the food industry or as probiotics in animal production, and that ERIC-PCR is a reliable tool to be used for molecular genetic profiling of potential probiotic enterococci.Item Design of Lactococcus lactis Strains Producing Garvicin A and/or Garvicin Q, Either Alone or Together with Nisin A or Nisin Z and High Antimicrobial Activity against Lactococcus garvieae(Foods, 2023) Feito Hermida, Javier; Araújo, Carlos; Arbulu, Sara; Contente, Diogo; Gómez Sala, Beatriz; Díaz-Formoso, Lara; Muñoz Atienza, Estefanía; Borrero Del Pino, Juan; Cintas Izarra, Luis Miguel; Hernández Cruza, Pablo ElpidioLactococcus garvieae is a main ichthyopathogen in rainbow trout (Oncorhynchus mykiss, Walbaum) farming, although bacteriocinogenic L. garvieae with antimicrobial activity against virulent strains of this species have also been identified. Some of the bacteriocins characterized, such as garvicin A (GarA) and garvicin Q (GarQ), may show potential for the control of the virulent L. garvieae in food, feed and other biotechnological applications. In this study, we report on the design of Lactococcus lactis strains that produce the bacteriocins GarA and/or GarQ, either alone or together with nisin A (NisA) or nisin Z (NisZ). Synthetic genes encoding the signal peptide of the lactococcal protein Usp45 (SPusp45), fused to mature GarA (lgnA) and/or mature GarQ (garQ) and their associated immunity genes (lgnI and garI, respectively), were cloned into the protein expression vectors pMG36c, which contains the P32 constitutive promoter, and pNZ8048c, which contains the inducible PnisA promoter. The transformation of recombinant vectors into lactococcal cells allowed for the production of GarA and/or GarQ by L. lactis subsp. cremoris NZ9000 and their co-production with NisA by Lactococcus lactis subsp. lactis DPC5598 and L. lactis subsp. lactis BB24. The strains L. lactis subsp. cremoris WA2-67 (pJFQI), a producer of GarQ and NisZ, and L. lactis subsp. cremoris WA2-67 (pJFQIAI), a producer of GarA, GarQ and NisZ, demonstrated the highest antimicrobial activity (5.1- to 10.7-fold and 17.3- to 68.2-fold, respectively) against virulent L. garvieae strains.- Project number: 25
Item Hacia un massive online open course (MOOC) para el aprendizaje de la higiene y seguridad alimentarias(2021) Borrero Del Pino, Juan; Chaparro Domínguez, María De Los Ángeles; Cintas Izarra, Luis Miguel; Contente de Matos, Diogo da Silva Serra; Feito Hermida, Javier; Hernández Cruza, Pablo Elpidio; Marín Martínez, María; Muñoz Atienza, Estefanía; Martín De Santos, María Del Rosario; Peña Vidal, Nuria; Gómez Sala, Beatriz; Álvarez López, Alberto; Celorrio de Ochoa, David; Alache Manchado, Beatriz; Ávila Alonso, Óscar; Beltrán Crespo, Antonio; Blanco Palmero, Felipe A.; Cabrales Miró-Granada, Ana; Carabante Oliver, Natalia; Carmona Agraz, Ana; Carpintero Cagigas, Ana; Casarrubios Guío, Pilar; Escobar Sáez, Daniel; Palenzuela González, Cristina; Romero Domingo, Sandra Item Production of Pumilarin and a Novel Circular Bacteriocin, Altitudin A, by Bacillus altitudinis ECC22, a Soil-Derived Bacteriocin Producer(International Journal of Molecular Sciences, 2024) Lafuente, Irene; Sevillano, Ester; Peña Vidal, Nuria; Cuartero, Alicia; Hernández Cruza, Pablo Elpidio; Cintas Izarra, Luis Miguel; Muñoz Atienza, Estefanía; Borrero Del Pino, JuanThe rise of antimicrobial resistance poses a significant global health threat, necessitating urgent efforts to identify novel antimicrobial agents. In this study, we undertook a thorough screening of soil-derived bacterial isolates to identify candidates showing antimicrobial activity against Gram-positive bacteria. A highly active antagonistic isolate was initially identified as Bacillus altitudinis ECC22, being further subjected to whole genome sequencing. A bioinformatic analysis of the B. altitudinis ECC22 genome revealed the presence of two gene clusters responsible for synthesizing two circular bacteriocins: pumilarin and a novel circular bacteriocin named altitudin A, alongside a closticin 574-like bacteriocin (CLB) structural gene. The synthesis and antimicrobial activity of the bacteriocins, pumilarin and altitudin A, were evaluated and validated using an in vitro cell-free protein synthesis (IV-CFPS) protocol coupled to a split-intein-mediated ligation procedure, as well as through their in vivo production by recombinant E. coli cells. However, the IV-CFPS of CLB showed no antimicrobial activity against the bacterial indicators tested. The purification of the bacteriocins produced by B. altitudinis ECC22, and their evaluation by MALDI-TOF MS analysis and LC-MS/MS-derived targeted proteomics identification combined with massive peptide analysis, confirmed the production and circular conformation of pumilarin and altitudin A. Both bacteriocins exhibited a spectrum of activity primarily directed against other Bacillus spp. strains. Structural three-dimensional predictions revealed that pumilarin and altitudin A may adopt a circular conformation with five- and four-α-helices, respectively.Item Antimicrobial Activity, Genetic Relatedness, and Safety Assessment of Potential Probiotic Lactic Acid Bacteria Isolated from a Rearing Tank of Rotifers (Brachionus plicatilis) Used as Live Feed in Fish Larviculture(Animals, 2024) Contente, Diogo; Díaz Formoso, Lara; Feito Hermida, Javier; Gómez Sala, Beatriz; Costas, Damián; Hernández Cruza, Pablo Elpidio; Muñoz Atienza, Estefanía; Borrero Del Pino, Juan; Poeta, Patrícia; Cintas Izarra, Luis MiguelAquaculture is a rapidly expanding agri-food industry that faces substantial economic losses due to infectious disease outbreaks, such as bacterial infections. These outbreaks cause disruptions and high mortalities at various stages of the rearing process, especially in the larval stages. Probiotic bacteria are emerging as promising and sustainable alternative or complementary strategies to vaccination and the use of antibiotics in aquaculture. In this study, potential probiotic candidates for larviculture were isolated from a rotifer-rearing tank used as the first live feed for turbot larvae. Two Lacticaseibacillus paracasei and two Lactiplantibacillus plantarum isolates were selected for further characterization due to their wide and strong antimicrobial activity against several ichthyopathogens, both Gram-positive and Gram-negative. An extensive in vitro safety assessment of these four isolates revealed the absence of harmful traits, such as acquired antimicrobial resistance and other virulence factors (i.e., hemolytic and gelatinase activities, bile salt deconjugation, and mucin degradation, as well as PCR detection of biogenic amine production). Moreover, Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) analyses unveiled their genetic relatedness, revealing two divergent clusters within each species. To our knowledge, this work reports for the first time the isolation and characterization of Lactic Acid Bacteria (LAB) with potential use as probiotics in aquaculture from rotifer-rearing tanks, which have the potential to optimize turbot larviculture and to introduce novel microbial management approaches for a sustainable aquaculture.Item Antimicrobial activity, antibiotic susceptibility and virulence factors of Lactic Acid Bacteria of aquatic origin intended for use as probiotics in aquaculture(BMC Microbiology, 2013) Muñoz Atienza, Estefanía; Gómez Sala, Beatriz; Araújo, Carlos; Campanero, Cristina; del Campo, Rosa; Hernández Cruza, Pablo Elpidio; Herranz Sorribes, Carmen; Cintas Izarra, Luis MiguelBackground. The microorganisms intended for use as probiotics in aquaculture should exert antimicrobial activity and be regarded as safe not only for the aquatic hosts but also for their surrounding environments and humans. The objective of this work was to investigate the antimicrobial/bacteriocin activity against fish pathogens, the antibiotic susceptibility, and the prevalence of virulence factors and detrimental enzymatic activities in 99 Lactic Acid Bacteria (LAB) (59 enterococci and 40 non-enterococci) isolated from aquatic animals regarded as human food. Results. These LAB displayed a broad antimicrobial/bacteriocin activity against the main Gram-positive and Gram-negative fish pathogens. However, particular safety concerns based on antibiotic resistance and virulence factors were identified in the genus Enterococcus (86%) (Enterococcus faecalis, 100%; E. faecium, 79%). Antibiotic resistance was also found in the genera Weissella (60%), Pediococcus (44%), Lactobacillus (33%), but not in leuconostocs and lactococci. Antibiotic resistance genes were found in 7.5% of the non-enterococci, including the genera Pediococcus (12.5%) and Weissella (6.7%). One strain of both Pediococcus pentosaceus and Weissella cibaria carried the erythromycin resistance gene mef(A/E), and another two P. pentosaceus strains harboured lnu(A) conferring resistance to lincosamides. Gelatinase activity was found in E. faecalis and E. faecium (71 and 11%, respectively), while a low number of E. faecalis (5%) and none E. faecium exerted hemolytic activity. None enterococci and non-enterococci showed bile deconjugation and mucin degradation abilities, or other detrimental enzymatic activities. Conclusions. To our knowledge, this is the first description of mef(A/E) in the genera Pediococcus and Weissella, and lnu(A) in the genus Pediococcus. The in vitro subtractive screening presented in this work constitutes a valuable strategy for the large-scale preliminary selection of putatively safe LAB intended for use as probiotics in aquaculture.