• The ability of Listeria monocytogenes to form biofilm on surfaces relevant to the mushroom production environment

      Dygico, Lionel Kenneth; Gahan, Cormac G.M.; Grogan, Helen; Burgess, Catherine; Department of Agriculture, Food and the Marine; 14F881 (Elsevier, 2020-03-16)
      Due to its ubiquitous nature, Listeria monocytogenes is a threat to all fresh fruits and vegetables, including mushrooms, which are Ireland's largest horticultural crop. Although fresh cultivated mushrooms (Agaricus bisporus) have not been previously linked with listeriosis outbreaks, the pathogen still poses a threat to the industry, particularly due to its ability to form biofilms. This threat is highlighted by the multiple recalls of mushroom products caused by L. monocytogenes contamination and by previous studies demonstrating that L. monocytogenes is present in the mushroom production environment. In this study, the biofilm formation potential of L. monocytogenes strains isolated from the mushroom production environment was investigated on materials and at temperatures relevant to mushroom production. A preliminary assessment of biofilm formation of 73 mushroom industry isolates was undertaken using a crystal violet assay on polystyrene microtitre plates. The biofilm formation of a subset (n = 7) of these strains was then assessed on twelve different materials, including materials that are representative of the materials commonly found in the mushroom production environments, using the CDC biofilm reactor. Vertical scanning interferometry was used to determine the surface roughness of the chosen materials. All the strains tested using the CDC biofilm reactor were able to form biofilms on the different surfaces tested but material type was found to be a key determining factor on the levels of biofilm formed. Stainless steel, aluminium, rubber, polypropylene and polycarbonate were all able to support biofilm levels in the range of 4–4.9 log10 CFU/cm2, for seven different L. monocytogenes strains. Mushroom industry-specific materials, including growing nets and tarpaulins, were found to support biofilms levels between 4.7 and 6.7 log10 CFU/cm2. Concrete was found to be of concern as it supported 7.7 log10 CFU/cm2 of biofilm for the same strains; however, sealing the concrete resulted in an approximately 2-log reduction in biofilm levels. The surface roughness of the materials varied greatly between the materials (0.7–3.5 log10 Ra) and was found to have a positive correlation with biofilm formation (rs = 0.573) although marginally significant (P = 0.051). The results of this study indicate that L. monocytogenes can readily form biofilms on mushroom industry relevant surfaces, and additionally identifies surfaces of specific concern, where rigorous cleaning and disinfection is required.
    • The ability of Listeria monocytogenes to form biofilm on surfaces relevant to the mushroom production environment

      Dygico, Lionel Kenneth; Gahan, Cormac G.M.; Grogan, Helen; Burgess, Catherine; Department of Agriculture, Food and the Marine; 14F881 (Elsevier BV, 2020-03-16)
      Due to its ubiquitous nature, Listeria monocytogenes is a threat to all fresh fruits and vegetables, including mushrooms, which are Ireland's largest horticultural crop. Although fresh cultivated mushrooms (Agaricus bisporus) have not been previously linked with listeriosis outbreaks, the pathogen still poses a threat to the industry, particularly due to its ability to form biofilms. This threat is highlighted by the multiple recalls of mushroom products caused by L. monocytogenes contamination and by previous studies demonstrating that L. monocytogenes is present in the mushroom production environment. In this study, the biofilm formation potential of L. monocytogenes strains isolated from the mushroom production environment was investigated on materials and at temperatures relevant to mushroom production. A preliminary assessment of biofilm formation of 73 mushroom industry isolates was undertaken using a crystal violet assay on polystyrene microtitre plates. The biofilm formation of a subset (n = 7) of these strains was then assessed on twelve different materials, including materials that are representative of the materials commonly found in the mushroom production environments, using the CDC biofilm reactor. Vertical scanning interferometry was used to determine the surface roughness of the chosen materials. All the strains tested using the CDC biofilm reactor were able to form biofilms on the different surfaces tested but material type was found to be a key determining factor on the levels of biofilm formed. Stainless steel, aluminium, rubber, polypropylene and polycarbonate were all able to support biofilm levels in the range of 4–4.9 log10 CFU/cm2, for seven different L. monocytogenes strains. Mushroom industry-specific materials, including growing nets and tarpaulins, were found to support biofilms levels between 4.7 and 6.7 log10 CFU/cm2. Concrete was found to be of concern as it supported 7.7 log10 CFU/cm2 of biofilm for the same strains; however, sealing the concrete resulted in an approximately 2-log reduction in biofilm levels. The surface roughness of the materials varied greatly between the materials (0.7–3.5 log10 Ra) and was found to have a positive correlation with biofilm formation (rs = 0.573) although marginally significant (P = 0.051). The results of this study indicate that L. monocytogenes can readily form biofilms on mushroom industry relevant surfaces, and additionally identifies surfaces of specific concern, where rigorous cleaning and disinfection is required.
    • Acrylamide formation in potato products

      Brunton, Nigel; Gormley, Ronan T.; Butler, Francis; Cummins, Enda; Danaher, Martin; O'Keeffe, Michael (Teagasc, 2006-08)
      Acrylamide, a substance classified as a potential carcinogen, occurs in heated starchy foods at concentrations many times in excess of levels permitted in drinking water. Early surveys indicated that levels of acrylamide in potato products such as French fries and potato crisps were the highest of the foodstuffs investigated. The present project addressed this issue by determining levels of acrylamide precursors (asparagine and reducing sugars) in raw potatoes and levels of acrylamide in (i) potato products from different storage regimes, (ii) spot-sampled potatoes purchased from a local supermarket, (iii) samples that received pre-treatments and were fried at different temperatures and (iv) French fries reheated in different ovens.A risk assessment of the estimated acrylamide intake from potato products for various cohorts of the Irish population was also conducted.
    • Antibiotic resistance in foodborne pathogens

      Duffy, Geraldine; Walsh, Ciara (Teagasc, 2005-02)
      Wide-spread antibiotic resistance among bacterial pathogens is now a serious public health issue and multi-antibiotic resistance has been reported in many foodborne pathogens including Salmonella and E. coli.
    • Antimicrobial activity of natural compounds against listeria spp. and their effects on sensory attributes in salmon (Salmo salar) and cod (Gadus morhua)

      Pedrós-Garrido, Selene; Clemente, Isabel; Calanche, J. B.; Condón-Abanto, Santiago; Beltrán, J. A.; Lyng, J. G.; Brunton, N.; Bolton, Declan; Whyte, Paul; Department of Agriculture, Food and the Marine; et al. (Elsevier, 2019-07-15)
      The application of natural preservatives on fresh fish has potential to extend shelf-life. In the present study, 8 essential oils (EOs) (lemon, lemongrass, lime, garlic, onion, oregano, thyme and rosemary) and 3 organic acids (OAs) (ascorbic, citric and lactic) were evaluated. The antimicrobial activity of these compounds was tested in-vitro against four confirmed Listeria spp. isolated from retail skin-packed salmon and cod. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were established for each compound. Then, a sensory evaluation was performed by a panel of ‘expert assessors’ on cooked fish treated with all of the OAs and any 4 EOs with a MIC <0.8%. A series of descriptors were assigned to characterize the combination of each compound with cooked salmon or cod. The highest antimicrobial effect against all Listeria spp. was observed for lactic acid (0.31–2.5%), but treatment with this compound resulted in the development of organoleptically unacceptable changes in salmon or cod. The most acceptable OAs for salmon and cod were ascorbic acid (1.25%) and citric acid (0.63%) respectively, which were shown to enhance certain organoleptic characteristics. The most effective EO against all Listeria strains evaluated was oregano oil (0.2%) and it was considered suitable as a treatment for salmon. In contrast, none of the EOs tested was organoleptically acceptable in combination with cod because of their strong odours and flavours that masked the fresh attributes associated with this fish.
    • Antimicrobial antagonists against food pathogens; a bacteriocin perspective

      O'Connor, Paula M.; Ross, R Paul; Hill, Colin; Cotter, Paul D.; Science Foundation Ireland; 12/RC/2273 (Elsevier, 03/02/2015)
      Efforts are continuing to find novel bacteriocins with enhanced specificity and potency. Traditional plating techniques are still being used for bacteriocin screening studies, however, the availability of ever more bacterial genome sequences and the use of in silico gene mining tools have revealed novel bacteriocin gene clusters that would otherwise have been overlooked. Furthermore, synthetic biology and bioengineering-based approaches are allowing scientists to harness existing and novel bacteriocin gene clusters through expression in different hosts and by enhancing functionalities. The same principles apply to bacteriocin producing probiotic cultures and their application to control pathogens in the gut. We can expect that the recent developments on bacteriocins from Lactic Acid Bacteria (LAB) described here will contribute greatly to increased commercialisation of bacteriocins in food systems.
    • Antimicrobial Resistance Determinants Circulating among Thermophilic Campylobacter Isolates Recovered from Broilers in Ireland Over a One-Year Period

      Lynch, Caoimhe T.; Lynch, Helen; Burke, Sarah; Hawkins, Kayleigh; Buttimer, Colin; McCarthy, Conor; Egan, John; Whyte, Paul; Bolton, Declan; Coffey, Aidan; et al. (MDPI AG, 2020-06-08)
      Campylobacteriosis is the leading cause of human bacterial gastroenteritis, very often associated with poultry consumption. Thermophilic Campylobacter (Campylobacter jejuni and Campylobacter coli) isolates (n = 158) recovered from broiler neck skin and caecal contents in Ireland over a one-year period, resistant to at least one of three clinically relevant antimicrobial classes, were screened for resistance determinants. All ciprofloxacin-resistant isolates (n = 99) harboured the C257T nucleotide mutation (conferring the Thr-86-Ile substitution) in conjunction with other synonymous and nonsynonymous mutations, which may have epidemiological value. The A2075G nucleotide mutation and amino acid substitutions in L4 and L22 were detected in all erythromycin-resistant isolates (n = 5). The tetO gene was detected in 100% (n = 119) of tetracycline-resistant isolates and three of which were found to harbour the mosaic tetracycline resistance gene tetO/32/O. Two streptomycin-resistant C. jejuni isolates (isolated from the same flock) harboured ant(6)-Ib, located in a multidrug resistance genomic island, containing aminoglycoside, streptothricin (satA) and tetracycline resistance genes (truncated tetO and mosaic tetO/32/O). The ant(6)-Ie gene was identified in two streptomycin-resistant C. coli isolates. This study highlights the widespread acquisition of antimicrobial resistance determinants among chicken-associated Campylobacter isolates, through horizontal gene transfer or clonal expansion of resistant lineages. The stability of such resistance determinants is compounded by the fluidity of mobile genetic element.
    • Automated detection and characterisation of foodborne pathogens

      Duffy, Geraldine; O'Hanlon, Karen; Catarame, Terese; Smyth, Davida S.; McCann, Máiréad (Teagasc, 2007-06)
      This study focused on the development of molecular tools for the rapid detection and characterisation of food-borne pathogens including Verocytotoxigenic Escherichia coli (VTEC) (serotypes O157, O26 and O111) and Salmonella spp. The study involved the development of enrichment systems and the identification of unique genetic targets in these pathogens which could be amplified and detected by Real Time Polymerase Chain Reaction (PCR).
    • Bacterial resistance to antibiotic alternatives: a wolf in sheep’s clothing?

      Willing, Benjamin P; Pepin, Deanna M; Marcolla, Camila S; Forgie, Andrew J; Diether, Natalie E; Bourrie, Benjamin C T (Oxford University Press, 2018-04-28)
      Implications • Substantial pressure to reduce antibiotic use has necessitated the development of antibiotic alternatives. However, relatively little consideration has been given to the development of resistance to these alternatives. • Whether we come up with antibiotic alternatives that are bacteriocidal or inhibitory, bacteria will continue to adapt and evolve. • Some antibiotic alternatives support the development of antibiotic resistance necessitating caution. • There are opportunities to optimize antibiotic alternative effectiveness as well as to minimize the development of resistance mechanisms.
    • Bactofencin A, a New Type of Cationic Bacteriocin with Unusual Immunity

      O'Shea, Eileen F.; O'Connor, Paula M.; O'Sullivan, Orla; Cotter, Paul D.; Ross, R Paul; Hill, Colin; Department of Agriculture, Food and the Marine, Ireland; Science Foundation Ireland; 04R; 07/CE/B1368 (American Society for Microbiology, 29/10/2013)
      Bacteriocin production is an important probiotic trait of intestinal bacteria. In this study, we identify a new type of bacteriocin, bactofencin A, produced by a porcine intestinal isolate Lactobacillus salivarius DPC6502, and assess its potency against pathogenic species including Staphylococcus aureus and Listeria monocytogenes. Genome sequencing of the bacteriocin producer revealed bfnA, which encodes the mature and highly basic (pI 10.59), 22-amino-acid defensin-like peptide. Matrixassisted laser desorption ionization–time of flight (MALDI-TOF) mass spectral analysis determined that bactofencin A has a molecular mass of 2,782 Da and contains two cysteine residues that form an intramolecular disulfide bond. Although an ABC transporter and transport accessory protein were also present within the bacteriocin gene cluster, a classical bacteriocin immunity gene was not detected. Interestingly, a dltB homologue was identified downstream of bfnA. DltB is usually encoded within the dlt operon of many Gram-positive bacteria. It is responsible for D-alanylation of teichoic acids in the cell wall and has previously been associated with bacterial resistance to cationic antimicrobial peptides. Heterologous expression of this gene conferred bactofencin A-specific immunity on sensitive strains of L. salivarius and S. aureus (although not L. monocytogenes), establishing its role in bacteriocin immunity. An analysis of the distribution of bfnA revealed that it was present in four additional isolates derived from porcine origin and absent from five human isolates, suggesting that its distribution is host specific. Given its novelty, we anticipate that bactofencin A represents the prototype of a new class of bacteriocins characterized as being cationic, with a DltB homologue providing a cognate immunity function.
    • Benzimidazole carbamate residues in milk: Detection by Surface Plasmon Resonance-biosensor, using a modified QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method for extraction

      Keegan, Jemma; Whelan, Michelle; Danaher, Martin; Crooks, Steven; Sayers, Riona; Anastasio, Aniello; Elliott, C.; Brandon, David; Furey, A.; O'Kennedy, Richard (Elsevier, 2009-09-26)
      A surface plasmon resonance (SPR) biosensor screening assay was developed and validated to detect 11 benzimidazole carbamate (BZT) veterinary drug residues in milk. The polyclonal antibody used was raised in sheep against a methyl 5(6)-[(carboxypentyl)-thio]-2-benzimidazole carbamate protein conjugate. A sample preparation procedure was developed using a modified QuEChERS method. BZT residues were extracted from milk using liquid extraction/partition with a dispersive solid phase extraction clean-up step. The assay was validated in accordance with the performance criteria described in 2002/657/EC. The limit of detection of the assay was calculated from the analysis of 20 known negative milk samples to be 2.7 μg kg−1. The detection capability (CCβ) of the assay was determined to be 5 μg kg−1 for 11 benzimidazole residues and the mean recovery of analytes was in the range 81–116%. A comparison was made between the SPR-biosensor and UPLC–MS/MS analyses of milk samples (n = 26) taken from cows treated different benzimidazole products, demonstrating the SPR-biosensor assay to be fit for purpose.
    • Bioactivity in Whey Proteins Influencing Energy Balance

      McAllan, Liam; Cotter, Paul D.; Roche, Helen M.; Korpela, Riitta; Nilaweera, Kanishka (OMICS Publishing Group, 30/03/2012)
      Obesity develops due to energy (food) intake exceeding energy expenditure. Nutrients that reduce the positive energy balance are thus being considered as therapies to combat obesity. Here, we review the literature related to the physiological, cellular and endocrine effects of intake of whey proteins, namely α-lactalbumin, β-lactoglobulin, glycomacropeptide and lactoferrin. Moreover, we discuss how dietary composition and obesity may influence whey protein effects on the above parameters. Evidence suggests that intake of whey proteins causes a decrease in energy intake, increase in energy expenditure, influence insulin sensitivity and glucose homeostasis and alter lipid metabolism in the adipose, liver and muscle. These physiological changes are accompanied by alterations in the plasma levels of energy balance related hormones (cholecystokinin, ghrelin, insulin and glucagon-like peptide-1) and the expression of catabolic and anabolic genes in the above tissue in the direction to cause a negative energy balance.
    • A case of bovine raw milk contamination with Listeria monocytogenes

      Hunt, Karen; Drummond, Niall; Murphy, Mary; Butler, Francis; Buckley, James F.; Jordan, Kieran; Teagasc Walsh Fellowship Programme; Department of Agriculture, Food and the Marine, Ireland; European Union (Biomed Central, 06/07/2012)
      During routine sampling of bulk raw milk on a dairy farm, the pathogenic bacteria Listeria monocytogenes was found to be a contaminant, at numbers < 100 cfu/ml. A strain with an indistinguishable pulsed-field gel electrophoresis pattern was isolated from the bulk milk two months later. Environmental swabs taken at the dairy environment were negative for the presence of L. monocytogenes, indicating a possible case of excretion of the L. monocytogenes directly into the milk. Milk samples were collected from the individual cows and analysed, resulting in the identification of L. monocytogenes excretion (at 280 cfu/ml) from one of the 4 mammary quarters of one dairy cow out of 180. When the infected cow was isolated from the herd, no L. monocytogenes was detected from the remaining herd. The pulsed-field gel electrophoresis pattern of the strain from the individual cow was indistinguishable from that originally isolated from the bulk milk. The infected cow did not show any clinical signs of disease, nor did the appearance of the milk have any physical abnormalities. Antibiotic treatment of the infected mammary quarter was found to be ineffective. This study shows that there can be risks associated with direct contamination of raw milk with L. monocytogenes.
    • Comparative analysis of Salmonella susceptibility and tolerance to the biocide chlorhexidine identifies a complex cellular defense network

      Condell, Orla; Power, Karen A.; Handler, Kristian; Finn, Sarah; Sheridan, Aine; Sergeant, Kjell; Renaut, Jenny; Burgess, Catherine; Hinton, Jay C.D.; Nally, Jarlath E.; et al. (Frontiers Media SA, 2014-08-01)
      Chlorhexidine is one of the most widely used biocides in health and agricultural settings as well as in the modern food industry. It is a cationic biocide of the biguanide class. Details of its mechanism of action are largely unknown. The frequent use of chlorhexidine has been questioned recently, amidst concerns that an overuse of this compound may select for bacteria displaying an altered susceptibility to antimicrobials, including clinically important anti-bacterial agents. We generated a Salmonella enterica serovar Typhimurium isolate (ST24CHX) that exhibited a high-level tolerant phenotype to chlorhexidine, following several rounds of in vitro selection, using sub-lethal concentrations of the biocide. This mutant showed altered suceptibility to a panel of clinically important antimicrobial compounds. Here we describe a genomic, transcriptomic, proteomic, and phenotypic analysis of the chlorhexidine tolerant S. Typhimurium compared with its isogenic sensitive progenitor. Results from this study describe a chlorhexidine defense network that functions in both the reference chlorhexidine sensitive isolate and the tolerant mutant. The defense network involved multiple cell targets including those associated with the synthesis and modification of the cell wall, the SOS response, virulence, and a shift in cellular metabolism toward anoxic pathways, some of which were regulated by CreB and Fur. In addition, results indicated that chlorhexidine tolerance was associated with more extensive modifications of the same cellular processes involved in this proposed network, as well as a divergent defense response involving the up-regulation of additional targets such as the flagellar apparatus and an altered cellular phosphate metabolism. These data show that sub-lethal concentrations of chlorhexidine induce distinct changes in exposed Salmonella, and our findings provide insights into the mechanisms of action and tolerance to this biocidal agent.
    • Comparative effect of different cooking methods on the physicochemical and sensory characteristics of high pressure processed marinated pork chops

      O'Neill, Ciara M.; Cruz-Romero, Malco C.; Duffy, Geraldine; Kerry, J. P.; Department of Agriculture, Food and the Marine; 11/F/031 (Elsevier, 2019-03-13)
      The objective of this study was to assess the effect of griddle and steam cooking on the physicochemical and sensory characteristics of high pressure processed (HPP) piri-piri marinated pork chops (MPC). Raw MPC that were HPP at 400 MPa had higher (P < 0.05) marinade absorption compared to untreated samples. After cooking, griddled MPC were significantly (P < 0.05) darker, less red, less yellow, tougher and had higher cook loss compared to steam cooked samples. The appearance of the griddled MPC was preferred while the texture, tenderness, juiciness and overall sensory acceptability (OSA) were preferred in steam cooked MPC. The increased marinade absorption in MPC that were HPP modified the fatty acid composition resulting in increased (P < 0.05) levels of oleic acid (C18:1c). Steam cooked MPC had a lower (P < 0.05) n-6: n-3 PUFA ratio and were preferred by the sensory panel compared to griddled MPC. Overall, from the cooking methods assessed steam cooking was the best cooking method for untreated and MPC that were HPP.
    • Comparative effect of different cooking methods on the physicochemical and sensory characteristics of high pressure processed marinated pork chops

      O'Neill, Ciara M.; Cruz-Romero, Malco C.; Duffy, Geraldine; Kerry, Joseph P.; Department of Agriculture, Fisheries & Food; 11/F/031 (Elsevier, 2019-03-13)
      The objective of this study was to assess the effect of griddle and steam cooking on the physicochemical and sensory characteristics of high pressure processed (HPP) piri-piri marinated pork chops (MPC). Raw MPC that were HPP at 400 MPa had higher (P < 0.05) marinade absorption compared to untreated samples. After cooking, griddled MPC were significantly (P < 0.05) darker, less red, less yellow, tougher and had higher cook loss compared to steam cooked samples. The appearance of the griddled MPC was preferred while the texture, tenderness, juiciness and overall sensory acceptability (OSA) were preferred in steam cooked MPC. The increased marinade absorption in MPC that were HPP modified the fatty acid composition resulting in increased (P < 0.05) levels of oleic acid (C18:1c). Steam cooked MPC had a lower (P < 0.05) n-6: n-3 PUFA ratio and were preferred by the sensory panel compared to griddled MPC. Overall, from the cooking methods assessed steam cooking was the best cooking method for untreated and MPC that were HPP. Industrial relevance: Processed meat manufacturers are constantly looking for new ways to increase yield, safety and shelf life of meat products. While high pressure processing (HPP) of raw meat has been shown to increase the safety and shelf life of these products; however, negative effects on the physicochemical characteristics of raw meat products have been reported. For example, HPP of raw meat products causes a whitening effect which may negatively affect consumers' acceptance of these products. In this study, we used a novel approach (a combination of HPP, marinade and a mix of organic acids Inbac™) which showed great potential not only for enhancing the yield of marinated pork chops but also enhancement of the sensory properties, safety and shelf life and particularly the piri-piri marinade masked the discoloration of raw pork meat caused by HPP. This study also provides consumers, retailers and caterers with information on how to best prepare HPP meat products and showed that steam cooked HPP marinated pork chops had the best physicochemical and sensory characteristics compared to griddled marinated pork chops.
    • Complete Genome Sequence of Clostridium estertheticum DSM 8809, a Microbe Identified in Spoiled Vacuum Packed Beef

      Zhongyi, Yu; Gunn, Lynda; Brennan, Evan; Reid, Rachael; Wall, Patrick G.; O Gaora, Peadar; Hurley, Daniel; Bolton, Declan; Fanning, Seamus; Department of Agriculture, Food and the Marine, Ireland (Frontiers, 2016-11-10)
      Blown pack spoilage (BPS) is a major issue for the beef industry. Etiological agents of BPS involve members of a group of Clostridium species, including Clostridium estertheticum which has the ability to produce gas, mostly carbon dioxide, under anaerobic psychotrophic growth conditions. This spore-forming bacterium grows slowly under laboratory conditions, and it can take up to 3 months to produce a workable culture. These characteristics have limited the study of this commercially challenging bacterium. Consequently information on this bacterium is limited and no effective controls are currently available to confidently detect and manage this production risk. In this study the complete genome of C. estertheticum DSM 8809 was determined by SMRT R sequencing. The genome consists of a circular chromosome of 4.7 Mbp along with a single plasmid carrying a potential tellurite resistance gene tehB and a Tn3- like resolvase-encoding gene tnpR. The genome sequence was searched for central metabolic pathways that would support its biochemical profile and several enzymes contributing to this phenotype were identified. Several putative antibiotic/biocide/metal resistance-encoding genes and virulence factors were also identified in the genome, a feature that requires further research. The availability of the genome sequence will provide a basic blueprint from which to develop valuable biomarkers that could support and improve the detection and control of this bacterium along the beef production chain.
    • Complete Genome Sequence of Clostridium estertheticum DSM 8809, a Microbe Identified in Spoiled Vacuum Packed Beef

      Yu, Zhongyi; Gunn, Lynda; Brennan, Evan; Reid, Rachael; Wall, Patrick G.; O Gaora, Peadar; Hurley, Daniel; Bolton, Declan; Fanning, Seamus; Department of Agriculture, Food and the Marine (Frontiers, 2016-11-11)
      Blown pack spoilage (BPS) is a major issue for the beef industry. Etiological agents of BPS involve members of a group of Clostridium species, including Clostridium estertheticum which has the ability to produce gas, mostly carbon dioxide, under anaerobic psychotrophic growth conditions. This spore-forming bacterium grows slowly under laboratory conditions, and it can take up to 3 months to produce a workable culture. These characteristics have limited the study of this commercially challenging bacterium. Consequently information on this bacterium is limited and no effective controls are currently available to confidently detect and manage this production risk. In this study the complete genome of C. estertheticum DSM 8809 was determined by SMRT® sequencing. The genome consists of a circular chromosome of 4.7 Mbp along with a single plasmid carrying a potential tellurite resistance gene tehB and a Tn3-like resolvase-encoding gene tnpR. The genome sequence was searched for central metabolic pathways that would support its biochemical profile and several enzymes contributing to this phenotype were identified. Several putative antibiotic/biocide/metal resistance-encoding genes and virulence factors were also identified in the genome, a feature that requires further research. The availability of the genome sequence will provide a basic blueprint from which to develop valuable biomarkers that could support and improve the detection and control of this bacterium along the beef production chain.
    • Complete Genome Sequences of vB_LmoS_188 and vB_LmoS_293, Two Bacteriophages with Specificity for Listeria monocytogenes Strains of Serotypes 4b and 4e

      Casey, A.; Kieran, Jordan; Coffey, Aidan; McAuliffe, Olivia; European Union; Safefood; PROMISE 265877; FOODSEG 266061 (American Society for Microbiology, 09/04/2015)
      Listeria monocytogenes is responsible for the rare disease listeriosis, which is associated with the consumption of contaminated food products. We report here the complete genome sequences of vB_LmoS_188 and vB_LmoS_293, phages isolated from environmental sources and that have host specificity for L. monocytogenes strains of the 4b and 4e serotypes.
    • Conserved redox-dependent DNA binding of ROXY glutaredoxins with TGA transcription factors

      Gutsche, Nora; Holtmannspötter, Michael; Maß, Lucia; O'Donoghue, Martin; Busch, Andrea; Lauri, Andrea; Schubert, Veit; Zachgo, Sabine; Deutsche Forschungsgemeinschaft; SPP 1710 ZA 259/7‐1; et al. (Wiley, 2017-12-14)
      The Arabidopsis thaliana CC‐type glutaredoxin (GRX) ROXY1 and the bZIP TGA transcription factor (TF) PERIANTHIA (PAN) interact in the nucleus and together regulate petal development. The CC‐type GRXs exist exclusively in land plants, and in contrast to the ubiquitously occurring CPYC and CGFS GRX classes, only the CC‐type GRXs expanded strongly during land plant evolution. Phylogenetic analyses show that TGA TFs evolved before the CC‐type GRXs in charophycean algae. MpROXY1/2 and MpTGA were isolated from the liverwort Marchantia polymorpha to analyze regulatory ROXY/TGA interactions in a basal land plant. Homologous and heterologous protein interaction studies demonstrate that nuclear ROXY/TGA interactions are conserved since the occurrence of CC‐type GRXs in bryophytes and mediated by a conserved ROXY C‐terminus. Redox EMSA analyses show a redox‐sensitive binding of MpTGA to the cis‐regulatory as‐1‐like element. Furthermore, we demonstrate that MpTGA binds together with MpROXY1/2 to this motif under reducing conditions, whereas this interaction is not observed under oxidizing conditions. Remarkably, heterologous complementation studies reveal a strongly conserved land plant ROXY activity, suggesting an ancestral role for CC‐type GRXs in modulating the activities of TGA TFs. Super‐resolution microscopy experiments detected a strong colocalization of ROXY1 with the active form of the RNA polymerase II in the nucleus. Together, these data shed new light on the function of ROXYs and TGA TFs and the evolution of redox‐sensitive transcription regulation processes, which likely contributed to adapt land plants to novel terrestrial habitats.