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Acetic and citric acids effect the type II secretion system and decrease the metabolic activities of salmon spoilage-related Rahnella aquatilis KM05

2024, Myszka, Kamila, Wolko, Łukasz, Borkowska, Monika

AbstractRahnella aquatilis causes seafoods to spoil by metabolizing sulfur-containing amino acids and/or proteins, producing H2S in products. The type II secretion system (T2SS) regulates the transport of proteases from the cytoplasm to the surrounding environment and promotes bacterial growth at low temperatures. To prevent premature fish spoilage, new solutions for inhibiting the T2SS of bacteria should be researched. In this study, global transcriptome sequencing was used to analyze the spoilage properties of R. aquatilis KM05. Two of the mapped genes/coding sequences (CDSs) were matched to the T2SS, namely, qspF and gspE, and four of the genes/CDSs, namely, ftsH, rseP, ptrA and pepN, were matched to metalloproteases or peptidases in R. aquatilis KM05. Subinhibitory concentrations of citric (18 µM) and acetic (41 µM) acids caused downregulation of T2SS-related genes (range from − 1.0 to -4.5) and genes involved in the proteolytic activities of bacteria (range from − 0.5 to -4.0). The proteolytic activities of R. aquatilis KM05 in vitro were reduced by an average of 40%. The in situ experiments showed the antimicrobial properties of citric and acetic acids against R. aquatilis KM05; the addition of an acidulant to salmon fillets limited microbial growth. Citric and acetic acids extend the shelf life of fish-based products and prevent food waste.

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Gallic and ferulic acids suppress proteolytic activities and volatile trimethylamine production in the food‐borne spoiler Rahnella aquatilis KM05

2023, Myszka, Kamila, Tomaś, Natalia, Wolko, Łukasz

AbstractBACKGROUNDRahnella aquatilis is a recognised microbial threat that alters the sensory properties of seafood. The high frequency with which R. aquatilis is isolated from fish has prompted a search for alternative preservatives. In the present study, in vitro and fish‐based ecosystem (raw salmon‐based medium) approaches were used to validate the antimicrobial effects of gallic (GA) and ferulic (FA) acids against R. aquatilis KM05. The results were compared with data describing the response of KM05 to sodium benzoate. Bioinformatics data of the whole genome were used to analyse the potential for fish spoilage by KM05 in detail, and the results revealed the main physiological characteristics that underlie reduced seafood quality.RESULTSIn the KM05 genome, the most abundantly enriched Gene Ontology terms were ‘metabolic process’, ‘organic substance metabolic process’ and ‘cellular process’. Through an evaluation of the Pfam annotations, 15 annotations were found to be directly involved in the proteolytic activity of KM05. Peptidase_M20 was the most abundantly represented (abundance value of 14060). Proteins representing the CutC family (abundance value of 427) indicated the potential for KM05 degradation of trimethyl‐amine‐N‐oxide. Subinhibitory concentrations of GA and FA suppressed the proteolytic activities of KM05 both in vitro and in RS medium by an average of 33–45%. These results were confirmed by quantitative real‐time PCR experiments, which also showed that the expression levels of genes involved in proteolytic activities and volatile trimethylamine production were also decreased.CONCLUSIONPhenolic compounds can be used as potential food additives for preventing quality deterioration of fish products. © 2023 Society of Chemical Industry.

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Expression patterns of candidate genes for the Lr46/Yr29 “slow rust” locus in common wheat (Triticum aestivum L.) and associated miRNAs inform of the gene conferring the Puccinia triticina resistance trait

2024, Spychała, Julia, Tomkowiak, Agnieszka, Noweiska, Aleksandra, Bobrowska, Roksana, Rychel-Bielska, Sandra, Bocianowski, Jan, Wolko, Łukasz, Kowalczewski, Przemysław Łukasz, Nowicki, Marcin, Kwiatek, Michał Tomasz

Leaf rust caused by Puccinia triticina (Pt) is one of the most impactful diseases causing substantial losses in common wheat (Triticum aestivum L.) crops. In adult plants resistant to Pt, a horizontal adult plant resistance (APR) is observed: APR protects the plant against multiple pathogen races and is distinguished by durable persistence under production conditions. The Lr46/Yr29 locus was mapped to chromosome 1B of common wheat genome, but the identity of the underlying gene has not been demonstrated although several candidate genes have been proposed. This study aimed to analyze the expression of nine candidate genes located at the Lr46/Yr29 locus and their four complementary miRNAs (tae-miR5384-3p, tae-miR9780, tae-miR9775, and tae-miR164), in response to Pt infection. The plant materials tested included five reference cultivars in which the molecular marker csLV46G22 associated with the Lr46/Yr29-based Pt resistance was identified, as well as one susceptible control cultivar. Biotic stress was induced in adult plants by inoculation with fungal spores under controlled conditions. Plant material was sampled before and at 6, 12, 24, 48 hours post inoculation (hpi). Differences in expression of candidate genes at the Lr46/Yr29 locus were analyzed by qRT-PCR and showed that the expression of the genes varied at the analyzed time points. The highest expression of Lr46/Yr29 candidate genes (Lr46-Glu1, Lr46-Glu2, Lr46-Glu3, Lr46-RLK1, Lr46-RLK2, Lr46-RLK3, Lr46-RLK4, Lr46-Snex, and Lr46-WRKY) occurred at 12 and 24 hpi and such expression profiles were obtained only for one candidate gene among the nine genes analyzed (Lr46-Glu2), indicating that it may be a contributing factor in the resistance response to Pt infection.

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Global transcriptome analysis of Pseudomonas aeruginosa NT06 response to potassium chloride, sodium lactate, sodium citrate, and microaerophilic conditions in a fish ecosystem

2024, Tomaś, Natalia, Myszka, Kamila, Wolko, Łukasz, Juzwa, Wojciech

Abstract Pseudomonas aeruginosa is an opportunistic pathogen that recently has been increasingly isolated from foods, especially from minimally processed fish-based products. Those are preserved by the addition of sodium chloride (NaCl) and packaging in a modified atmosphere. However, the current trends of minimizing NaCl content may result in an increased occurrence of P. aeruginosa. NaCl can be replaced with potassium chloride (KCl) or sodium salts of organic acids. Herein, we examined the antimicrobial effects of KCl, sodium lactate (NaL), sodium citrate (NaC), and sodium acetate (NaA) against P. aeruginosa NT06 isolated from fish. Transcriptome response of cells grown in medium imitating a fish product supplemented with KCl and KCl/NaL/NaC and maintained under microaerophilic conditions was analysed. Flow cytometry analysis showed that treatment with KCl and KCl/NaL/NaC resulted in changed metabolic activity of cells. In response to KCl and KCl/NaL/NaC treatment, genes related to cell maintenance, stress response, quorum sensing, virulence, efflux pump, and metabolism were differentially expressed. Collectively, our results provide an improved understanding of the response of P. aeruginosa to NaCl alternative compounds that can be implemented in fish-based products and encourage further exploration of the development of effective methods to protect foods against the P. aeruginosa, underestimate foodborne bacteria.