Strain C58 Research Articles

Identification of End-Binding 1 Protein as Novel α-4 Giardin-Binding Partners in Giardia lamblia Trophozoites.

Giardia lamblia (syn. G. intestinalis, G. duodenalis) is a primitive opportunistic protozoon, and one of the earliest differentiated eukaryotes. Despite its primitive nature, G. lamblia has a sophisticated cytoskeleton system, which is closely related to its proliferation and pathogenicity. Meanwhile, α giardin is a G. lamblia-specific cytoskeleton protein, which belongs to the annexin superfamily. Interestingly, G. lamblia has 21 annexin-like α giardins, i.e., more than higher eukaryotes. The functional differences among α giardin members are not fully understood. We took α-4 giardin, a member of α giardin family, as a research object. A morpholino-mediated knockdown experiment was performed to identify the effect of α-4 giardin on G. lamblia trophozoites biological traits. A yeast two-hybrid cDNA library of G. lamblia strain C2 trophozoites was screened for interaction partners of α-4 giardin. Co-immunoprecipitation and fluorescent colocalization confirmed the relationship between G. lamblia EB1 (gEB1) and α-4 giardin. α-4 Giardin could inhibit the proliferation and adhesion of G. lamblia trophozoites. In addition, it interacted with G. lamblia EB1 (gEB1). α-4 Giardin was involved in proliferation and adhesion in G. lamblia trophozoites, and EB1, a crucial roles in mitosis, was an interacting partner of α-4 giardin.

Streptococcus suis subtilisin-like serine proteases SspA-1 and SspA-2 interplay with complement C3a and C5a to facilitate bacterial immune evasion and infection

ABSTRACT Streptococcus suis (S. suis), a significant zoonotic bacterial pathogen impacting swine and human, is associated with severe systemic diseases such as streptococcal toxic shock-like syndrome, meningitis, septicaemia, and abrupt fatality. The multifaceted roles of complement components C5a and C3a extend to orchestrating inflammatory cells recruitment, oxidative burst induction, and cytokines release. Despite the pivotal role of subtilisin-like serine proteases in S. suis pathogenicity, their involvement in immune evasion remains underexplored. In the present study, we identify two cell wall-anchored subtilisin-like serine proteases in S. suis, SspA-1 and SspA-2, as binding partners for C3a and C5a. Through Co-Immunoprecipitation, Enzyme-Linked Immunosorbent and Far-Western Blotting Assays, we validate their interactions with the aforementioned components. However, SspA-1 and SspA-2 have no cleavage activity against complement C3a and C5a performed by Cleavage assay. Chemotaxis assays reveal that recombinant SspA-1 and SspA-2 effectively attenuate monocyte chemotaxis towards C3a and C5a. Notably, the ΔsspA-1, ΔsspA-1, and ΔsspA-1/2 mutant strains exhibit compromised survival in blood, and resistance of opsonophagocytosis, alongside impaired survival in blood and in vivo colonization compared to the parental strain SC-19. Critical insights from the murine and Galleria mellonella larva infection models further underscore the significance of sspA-1 in altering mortality rates. Collectively, our findings indicate that SspA-1 and SspA-2 are novel binding proteins for C3a and C5a, thereby shedding light on their pivotal roles in S. suis immune evasion and the pathogenesis.

Promoter Prediction in Agrobacterium tumefaciens Strain C58 by Using Artificial Intelligence Strategies.

Promoters are the genomic regions upstream of genes that RNA polymerase binds in order to initiate gene transcription. Understanding the regulation of gene expression depends on being able to identify promoters, because they are the most important component of gene expression. Agrobacterium tumefaciens (A. tumefaciens) strain C58 was the subject of this study with the goal of creating a machine learning-based model to predict promoters. In this study, nucleotide density (ND), k-mer, and one-hot were used to encode the promoter sequence. Support vector machine (SVM) on fivefold cross-validation with incremental feature selection (IFS) was used to optimize the generated features. These improved characteristics were then used to distinguish promoter sequences by feeding them into the random forest (RF) classifier. Tenfold cross-validation (CV) analysis revealed that the projected model has the ability to produce an accuracy of 84.22%.

Cross Flow Microfiltration System in Separating Fermented Nixtamal Corn for Preparation of Natural Folic Acid

A purification process of biomass of nixtamal yellow corn fermented by Rhizopus oligosporus strain C1 (FNC-A) and Rhizopus sp (FNC-B) was performed using a microfiltration (MF) membrane with pore size 0.15 µm installed in a crossflow microfiltration (CFMF) system to obtain retentate and permeate fractions as natural folic acid preparations. CFMF process was conducted at room temperature with a pump motor frequency (PMF) of 10 Hz (flow rate of 3.5 L/min) and a transmembrane pressure (TMP) of 4 bar for 0, 15, 30, 45, 60, 75, and 90 minutes. Based on optimum folic acid, the results showed that the best treatments of FNC-B and FNC-B were achieved at 90 min, resulting in a folic acid increase in retentate of FNC-A and FNC- B of 72.61 and 95.26%, reducing sugar of 20.41 and 170.93% (1.7-folds), total sugars of 155.51% (1.55-folds) and 426.76% (4.27-folds), and dissolved protein of 55.33 and 39.20%, and decrease in total solids of 88.91 and 91.64%, respectively, compared to initial biomass of FNC-A and FNC-B. The MFCF system effectively separated folic acid in retentates of FNC-A (33.77%) and FNC-B (95.27%) at the optimal condition. Folic acid monomers predominated the characteristics of FNC-A and FNC-B in optimum conditions with molecular weights of 442.10 and 442.18 Dalton, the average particle size of 38.31 μm and 37.97 μm, and distribution of particles at 10, 50, and 90% from the particle size 10.31, 26.32 and 81.55 μm, and 10.37, 28.04 and 76.09 μm, respectively.

Cross Flow Microfiltration System in Separating Fermented Nixtamal Corn for Preparation of Natural Folic Acid

A purification process of biomass of nixtamal yellow corn fermented by Rhizopus oligosporus strain C1 (FNC-A) and Rhizopus sp (FNC-B) was performed using a microfiltration (MF) membrane with pore size 0.15 µm installed in a crossflow microfiltration (CFMF) system to obtain retentate and permeate fractions as natural folic acid preparations. CFMF process was conducted at room temperature with a pump motor frequency (PMF) of 10 Hz (flow rate of 3.5 L/min) and a transmembrane pressure (TMP) of 4 bar for 0, 15, 30, 45, 60, 75, and 90 minutes. Based on optimum folic acid, the results showed that the best treatments of FNC-B and FNC-B were achieved at 90 min, resulting in a folic acid increase in retentate of FNC-A and FNC- B of 72.61 and 95.26%, reducing sugar of 20.41 and 170.93% (1.7-folds), total sugars of 155.51% (1.55-folds) and 426.76% (4.27-folds), and dissolved protein of 55.33 and 39.20%, and decrease in total solids of 88.91 and 91.64%, respectively, compared to initial biomass of FNC-A and FNC-B. The MFCF system effectively separated folic acid in retentates of FNC-A (33.77%) and FNC-B (95.27%) at the optimal condition. Folic acid monomers predominated the characteristics of FNC-A and FNC-B in optimum conditions with molecular weights of 442.10 and 442.18 Dalton, the average particle size of 38.31 μm and 37.97 μm, and distribution of particles at 10, 50, and 90% from the particle size 10.31, 26.32 and 81.55 μm, and 10.37, 28.04 and 76.09 μm, respectively.

O-acetyl-homoserine sulfhydrylase deficient Streptococcus suis serotype 2 strain SC19 becomes an avirulent strain and provides immune protection against homotype infection in mice

O-acetyl-homoserine sulfhydrylase (OAHS) is a pyridoxal 5’-phosphate-dependent enzyme involved in microbial methionine biosynthesis, which catalyzes the conversion of o-acetyl-homoserine (OAH) to homocysteine. In our previous study, we found that OAHS of Streptococcus suis serotype 2 (SS2) can interact with the porcine blood-brain barrier (BBB) model, but whether OAHS regulates the penetration of BBB during SS2 infection is still unclear. To explore the role of OAHS in SS2 infection, OAHS-deficient SS2 mutant strain (SC19-ΔOAHS) and gene complemental strain (SC19-cΔOAHS) were constructed. Compared to the parent strain, with the loss of oahs, the chain length of SC19-ΔOAHS was shortened, the virulence was significantly reduced, the survival rate of mice infected with SC19-ΔOAHS was obviously increased accompanied by the relieved clinical symptoms. And the survival ability of SC19-ΔOAHS in whole blood was also remarkably decreased. Interestingly, the adhesion of SC19-ΔOAHS to endothelial cells was markedly increased, but the deficiency of OAHS significantly inhibited the strain penetrating BBB both in vivo and in vitro. Most of these phenomena can be reversed by the complemental strain (SC19-cΔOAHS). Further study showed that the deficiency of OAHS severely reduced SC19-induced endothelial cell apoptosis, tight junctions (TJs) protein impairment and the expression of SS2 virulence factor Enolase (Eno), involved in the destruction of BBB. Additionally, SC19-ΔOAHS immunized mice were able to resist SC19 or JZLQ022 infection. In conclusion, we confirmed that OAHS promoted the pathogenicity by enhancing host’s BBB permeability and immune escape, and SC19- ΔOAHS is a potential live vaccine.

Resistance phenotype and genetic features of a heterogeneous vancomycin intermediate-resistant Staphylococcus aureus strain from an immunocompromised patient.

Strain C1 was successfully isolated from an immunosuppressed patient with persistent bacteremia, who had not previously been exposed to glycopeptide antibiotics. This strain was found to be a heterogeneous vancomycin intermediate-resistant Staphylococcus aureus (hVISA). It is noteworthy that, following a brief period of vancomycin treatment, strains C6, C8, and C9, which were obtained from blood and other body parts, exhibited a significant reduction in heterogeneity as determined by population analysis profile-area under the curve (PAP-AUC) detection. Genotyping analysis revealed that these bacterial strains belonged to the same SCCmecIVa-ST59-t437-agrI genotype and shared the same virulome and resistome. In this study, a comparative genomics analysis was conducted between strain C1 and strain N315 to identify potential hVISA-associated mutations. Ultimately, a total of 205 mutation sites in 19 candidate genes, likely associated with the hVISA phenotype, were identified.

Genome Sequence Resource of Cladosporium velox Strain C4 Causing Cotton Boll Disease in Xinjiang, China.

Cladosporium spp., as one of the largest and most heterogeneous genera of hyphomycetes, are widely distributed worldwide. This genus is usually adaptable to a wide variety of extreme environments. However, only 11 genomes of Cladosporium genus have been publicly released. From 2017, we found for the first time that Cladosporium velox could cause cotton boll disease and lead to stiffness and cracking boll in Xinjiang, China. Herein, we provide a high-quality reference genome for the C. velox strain C4 isolated from cotton boll in Xinjiang, China. The genome size and encoding gene number of the C. velox strain C4 and C. cucumerinum strain CCNX2, which was recently released and caused the cucumber scab, showed minor differences. This resource will contribute to future research that aims to elucidate the genetic basis of C. velox pathogenicity and could expand our knowledge of Cladosporium spp. genomic characteristics that will be valuable for the development of Cladosporium disease control measures.

Aerococcus viridans Phage Lysin AVPL Had Lytic Activity against Streptococcus suis in a Mouse Bacteremia Model

Streptococcus suis (S. suis) is a swine pathogen that can cause sepsis, meningitis, endocarditis, and other infectious diseases; it is also a zoonotic pathogen that has caused a global surge in fatal human infections. The widespread prevalence of multidrug-resistant S. suis strains and the decline in novel antibiotic candidates have necessitated the development of alternative antimicrobial agents. In this study, AVPL, the Aerococcus viridans (A. viridans) phage lysin, was found to exhibit efficient bactericidal activity and broad lytic activity against multiple serotypes of S. suis. A final concentration of 300 μg/mL AVPL reduced S. suis counts by 4–4.5 log10 within 1 h in vitro. Importantly, AVPL effectively inhibited 48 h S. suis biofilm formation and disrupted preformed biofilms. In a mouse model, 300 μg/mouse AVPL protected 100% of mice from infection following the administration of lethal doses of multidrug-resistant S. suis type 2 (SS2) strain SC19, reduced the bacterial load in different organs, and effectively alleviated inflammation and histopathological damage in infected mice. These data suggest that AVPL is a valuable candidate antimicrobial agent for treating S. suis infections.

The variations of native plasmids greatly affect the cell surface hydrophobicity of sphingomonads.

Microbial cell surface hydrophobicity (CSH) reflects nonspecific adhesion ability and affects various physiological processes, such as biofilm formation and pollutant biodegradation. Understanding the regulation mechanisms of CSH will contribute to illuminating microbial adaptation strategies and provide guidance for controlling CSH artificially to benefit humans. Sphingomonads, a common bacterial group with great xenobiotic-degrading ability, generally show higher CSH than typical Gram-negative bacteria, which plays a positive role in organic pollutant capture and cell colonization. This study verified that the variations of two native plasmids involved in synthesizing outer membrane proteins and polysaccharides greatly affected the CSH of sphingomonads. It is feasible to control their CSH by changing the plasmid copy number and sequences. Additionally, considering that plasmids are likely to evolve faster than chromosomes, the CSH of sphingomonads may evolve quickly to respond to environmental changes. Our results provide valuable insights into the CSH regulation and evolution of sphingomonads.

Transcriptomics reveals the mechanism of high-adhesive Bacillus subtilis inhibiting fresh fish spoilage

Aquatic products are highly susceptible to spoilage caused by adherence of spoilage bacteria during storage, so it is crucial to develop new microecological preservatives. The probiotic B. subtilis can achieve potential preservation effects by adhering to the fish gut to regulate the amount of probiotic colonization in the flora. Here, we demonstrated the surface adhesion characteristics of B. subtilis and the mechanism of adhesion-associated gene-mediated adhesion; the regulation of B. subtilis adhesion ability by nitrogen and carbon sources, and the application of tilapia preservation. Our results revealed 10 B. subtilis adhesion-related genes, among which nitrogen sources up-regulate the adhesion genes hag and motA, carbon sources up-regulate sinR and fliG. Based on flagellum-associated high adhesion strain C6 and high adhesion regulation of B. subtilis C6_H allowed the abundance of probiotic bacteria Bacillus, Rhodobacteraceae to increase, attenuate the content of spoilage flavor substances such as 1-Octen-3-ol, 15-Crown-5 and maintain the freshness of tilapia. The results helped to reveal the adhesion mechanism of B. subtilis and to develop and apply microecological preservatives with high quality adhesion ability.

Acquisition of Type I methyltransferase via horizontal gene transfer increases the drug resistance of Aeromonas veronii.

Aeromonas veronii is an opportunistic pathogen that affects both fish and mammals, including humans, leading to bacteraemia, sepsis, meningitis and even death. The increasing virulence and drug resistance of A. veronii are of significant concern and pose a severe risk to public safety. The Type I restriction-modification (RM) system, which functions as a bacterial defence mechanism, can influence gene expression through DNA methylation. However, little research has been conducted to explore its origin, evolutionary path, and relationship to virulence and drug resistance in A. veronii. In this study, we analysed the pan-genome of 233 A. veronii strains, and the results indicated that it was 'open', meaning that A. veronii has acquired additional genes from other species. This suggested that A. veronii had the potential to adapt and evolve rapidly, which might have contributed to its drug resistance. One Type I methyltransferase (MTase) and two complete Type I RM systems were identified, namely AveC4I, AveC4II and AveC4III in A. veronii strain C4, respectively. Notably, AveC4I was exclusive to A. veronii C4. Phylogenetic analysis revealed that AveC4I was derived from horizontal gene transfer from Thiocystis violascens and exchanged genes with the human pathogen Comamonas kerstersii. Single molecule real-time sequencing was applied to identify the motif methylated by AveC4I, which was unique and not recognized by any reported MTases in the REBASE database. We also annotated the functions and pathways of the genes containing the motif, revealing that AveC4I may control drug resistance in A. veronii C4. Our findings provide new insight on the mechanisms underlying drug resistance in pathogenic bacteria. By identifying the specific genes and pathways affected by AveC4I, this study may aid in the development of new therapeutic approaches to combat A. veronii infections.

Metabolic Profiling and Comparative Proteomic Insight in Respect of Amidases during Iprodione Biodegradation.

The fungicide iprodione (IPR) (3-(3,5-dichlorophenyl) N-isopropyl-2,4-dioxoimidazolidine-1-carboxamide) is a highly toxic compound. Although IPR has been restricted, it is still being applied in many places around the world, constituting an environmental risk. The biodegradation of IPR is an attractive option for reducing its residues. In this study, we isolated thirteen IPR-tolerant bacteria from a biopurification system designed to treat pesticides. A study of biodegradation using different strains was comparatively evaluated, and the best degradation rate of IPR was presented by Achromobacter sp. C1 with a half-life (T1/2) of 9 days. Based on a nano-LC-MS/MS analysis for the strains, proteins solely expressed in the IPR treatment were identified by highlighting the strain Achromobacter sp. C1, with 445 proteins primarily involved in the biosynthesis of secondary metabolites and microbial metabolism in diverse environments. Differentially expressed protein amidases were involved in six metabolic pathways. Interestingly, formamidase was inhibited while other cyclases, i.e., amidase and mandelamide hydrolase, were overexpressed, thereby minimizing the effect of IPR on the metabolism of strain C1. The dynamic changes in the protein profiles of bacteria that degrade IPR have been poorly studied; therefore, our results offer new insight into the metabolism of IPR-degrading microorganisms, with special attention paid to amidases.

Heavy-metal tolerant bacterial strains isolated from industrial sites and scrap yards in Kashmir, India.

Heavy metal pollution has posed a severe danger to environmental stability due to its high toxicity and lack of biodegradability. The present study deals with the appraisement of tolerance shown by various bacteria in varied copper and iron concentrations. Among the 20 isolates, four isolates, GN2, SC5, SC8, and SC10, exhibiting more significant iron and copper tolerance, were selected and identified by 16 S ribosomal ribonucleic acid (rRNA) gene sequence analysis as Pantoea agglomerans strain GN2, Pantoea sp. strain SC5, Bacillus sp. strain SC8 and Priestia aryabhattaistrain SC10. The minimum inhibitory concentration of molecularly identified strains revealed that P. agglomerans strain GN2 showed tolerance to iron sulfate and copper sulfate upto 600 and 400 µg/mL, whereas Bacillus sp. SC8 (OQ202165) showed tolerance of 700 and 250 µg/mL were tolerant to iron sulfate and copper sulfate up to 700 and 150 µg/mL, respectively. Pantoea sp. strain SC5 showed significant tolerance to both heavy metals. The isolates were further studied for their ability to grow at varying temperatures and pH ranges. Most of the isolates showed optimal growth at 37°C and pH 7. However, Pantoea sp. SC5 was competent to have prominent growth at 45°C and pH 8.0. Microbial remediation, which is eco-friendly, has proven the most effective method for bioremediation of heavy metal-contaminated environments. Using heavy metal-resistant bacteria for microbial remediation of iron and copper-contaminated environments could be a viable and valuable strategy. These isolates could also be used to decontaminate heavy metal-polluted agricultural soils.

Metabolic insights into how multifunctional microbial consortium enhances atrazine removal and phosphorus uptake at low temperature

Agricultural soils in the black soil region of northeast China often face negative stress due to low temperatures, pesticide contamination, and inadequate nutrient supply. In this study, a new cold-tolerant strain of Peribacillus simplex C1 (C1) was selectively isolated from atrazine contaminated soil. The artificially constructed microbial consortium (CPD) [C1, phosphorus-solubilizing bacterium Enterobacter sp. P1, and atrazine-degrading bacterium Acinetobacter lwoffii DNS32] demonstrated the most effective performance in enhancing atrazine degradation and phosphorus-solubilizing capacity when the initial inoculation ratio of 5:1:2 at 15 °C. CPD enhanced energy-related metabolic pathways and increased choline production to regulate bacterial adaptation to temperature decrease. Additionally, the strains could selectively utilize carbon sources (low molecular weight organic acids) or nitrogen sources (some metabolites of atrazine) provided by each other to enhance growth. Furthermore, strain C1 enhanced membrane fluidity through increased expression of the unsaturated fatty acids. Pot experiments demonstrated that CPD assisted soybean seedlings in resisting dual stresses of low temperature and atrazine contamination by inducing the expression of genes related to photosynthesis, membrane permeability, phosphorus response, and cold tolerance.

GmTOC1b negatively regulates resistance to Soybean mosaic virus

Soybean (Glycine max) is a major oil and feed crop worldwide. Soybean mosaic virus (SMV) is a globally occurring disease that severely reduces the yield and quality of soybean. Here, we characterized the role of the clock gene TIMING OF CAB EXPRESSION 1b (GmTOC1b) in the resistance of soybean to SMV. Homozygous Gmtoc1b mutants exhibited increased tolerance to SMV strain SC3 due to the activation of programmed cell death triggered by a hypersensitive response. Transcriptome deep sequencing and RT-qPCR analysis suggested that GmTOC1b likely regulates the expression of target genes involved in the salicylic acid (SA) signaling pathway. GmTOC1b binds to the promoter of GmWRKY40, which encodes a protein that activates the expression of SA-mediated defense-related genes. Moreover, we revealed that the GmTOC1bH1 haplotype, which confers increased tolerance to SMV, was artificially selected in improved cultivars from the Northern and Huang-Huai regions of China. Our results therefore identify a previously unknown SMV resistance component that could be deployed in the molecular breeding of soybean to enhance SMV resistance.

Isolation, Identification, and Application of Pigment-Producing Actinobacteria from Stingless Bee Hives for Handicraft Production

Actinobacteria are distributed in natural habitats and produce a vast variety of natural pigments that are different in colour shades and applied in textile industry and others. The extracted actinobacterial pigments are used as eco-friendly natural dyes and colour and non-toxic to living organisms and environments compared with chemical or synthetic colours. In order to produce actinobacterial pigments for handicraft making, eight strains of the pigmented producing actinobacteria were isolated from stingless bee (Tetragonilla collina Smith, 1857) hives. Based on deep colour shades presented, 4 actinobacterial strains were chosen to prepare various colour for handicrafts, including yellow, violet, green, and pink. To identify actinobacterial strains selected, phylogenetic identification was carried out. The phylogenetic results indicated that strain C2 phylogenetically shared the 16S rDNA sequence 99.6 % similarity with its closest species, namely Streptomyces cellulosae. Strain C4 had the phylogenetic relationship close to Streptomyces californicus (99.9% similarity of 16S rDNA). Strain E1 was a closest member belonged to Streptomyces chartreusis (99.9% 16S rDNA sequence similarity), and strain E2 was phylogenetically closely related to Streptomyces aureoversilis with 99.2% 16S rDNA sequence similarity. To produce the handicraft colour, we cultivated four actinobacterial strains on broken-milled rice, and actinobacterial pigments were extracted using ethyl acetate. The crude extracts obtained were mixed with white flower clay that was used for handicraft making, and the artificial clay flowers were made. This study is the first to report the use of natural colours obtained from pigment-producing actinobacteria in handicrafts in Thailand and other countries. Future research will explore the application of actinobacterial pigments in various fields, including fine arts, the ceramic industry, and others.

A distinct variant of the SzM protein of Streptococcus equi subsp. zooepidemicus recruits C1q independent of IgG binding and inhibits activation of the classical complement pathway

ABSTRACT Streptococcus equi subsp. zooepidemicus (SEZ) is a major equine pathogen that causes pneumonia, abortion, and polyarthritis. It can also cause invasive infections in humans. SEZ expresses the M-like protein SzM, which recruits host proteins such as fibrinogen to the bacterial surface. Equine SEZ strain C2, which binds only comparably low amounts of human fibrinogen in comparison to human SEZ strain C33, was previously shown to proliferate in equine and human blood. As the expression of SzM_C2 was necessary for survival in blood, this study investigated the working hypothesis that SzM_C2 inhibits complement activation through a mechanism other than fibrinogen and non-immune immunoglobulin binding. Loss-of-function experiments showed that SEZ C2, but not C33, binds C1q via SzM in IgG-free human plasma. Furthermore, SzM C2 expression is necessary for recruiting purified human or equine C1q to the bacterial surface. Flow cytometry analysis demonstrated that SzM expression in SEZ C2 is crucial for the significant reduction of C3b labelling in human plasma. Addition of human plasma to immobilized rSzM_C2 and immobilized aggregated IgG led to binding of C1q, but only the latter activated the complement system, as shown by the detection of C4 deposition. Complement activation induced by aggregated IgG was significantly reduced if human plasma was pre-incubated with rSzM_C2. Furthermore, rSzM_C2, but not rSzM_C33, inhibited the activation of the classical complement pathway in human plasma, as determined in an erythrocyte lysis experiment. In conclusion, the immunoglobulin-independent binding of C1q to SzM_C2 is associated with complement inhibition.

The Effect of Yeast, Sugar and Sulfur Dioxide on the Volatile Compounds in Wine

This study compares three yeast strains: two wild Saccharomyces cerevisiae strains (Sc1 and Sc5) and a commercial strain (Lc). The objective is to assess their fermentation efficiency and volatile compound production. The factors examined are yeast strain, initial sugar concentration of the must, and the presence of sulfur dioxide. Volatile aroma compounds, determined via GC–MS, were categorized into aromatic series based on aroma descriptors. Out of the volatile compounds analyzed, the yeast strain influenced 39, while sugar content and sulfur dioxide affected 16 and 23 compounds, respectively. Twelve compounds displayed odor activity values exceeding unity, with notable contributions from ethyl esters, β-damascenone, and β-ionone, impacting fruit, floral, and herbal aromatic series. Overall, the Sc1 yeast strain exhibited higher values in the aromatic series compared to the Lc strain. Multivariate analysis revealed that the Sc1 strain highlighted green fruit, citrus, and spice series, while the Lc strain stood out for smoky and herbal aromas. Cluster and principal component analyses emphasized that the aromatic composition of wines produced with wild yeast strains is more influenced by sulfur dioxide than initial sugar content, whereas the opposite holds true for the commercial strain. The key aroma series distinguishing between yeast strains were fruity, green fruit, and citrus for Sc1, and herbal, floral, and smoky for Lc. In conclusion, the Sc1 wild yeast strain showed similar fermentation behavior to the commercial strain, resulting in increased aroma compound presence. The distinctive aromatic profiles contributed by each strain enable winemakers to leverage this diversity and create wines that emphasize specific aromas.

Studying the metabolic factors that may impact the growth of co-cultured Listeria monocytogenes strains at low temperature

The simultaneous presence of more than one strains of Listeria monocytogenes in the same food product may affect the growth capacity of each strain. The present study evaluated the metabolites composition that may potentially influence the growth of individual L. monocytogenes strains in a dual strain composite. Based on previous studies, L. monocytogenes strains, C5 (4b) and 6179 (1/2a) were selected due to the remarkable interaction, which was observed during their co-culture. The selected strains were inoculated (2.0 - 3.0 log CFU/mL) in Tryptic Soy Broth with 0.6% Yeast Extract (TSB-YE) in single and two-strain cultures (1:1 strain ratio). Bacterial growth was assessed during storage at 7 °C, under aerobic conditions (AC). Their resistance to different antibiotics enabled the selective enumeration of each strain in the co-culture. After reaching stationary phase, single and dual cultures were centrifuged and filtered. The cell-free spent medium (CFSM) was either characterized by Fourier transform infrared (FTIR-ATR) spectrometry or re-inoculated, after the addition of concentrated TSB-YE (for nutrient replenishment), with single and two-strain cultures for the evaluation of growth under the influence of metabolites produced from the same singly and co-cultured strains in the different combinations of strains and CFSM origin (7 °C/AC) (n = 2x3). By the end of storage, singly-cultured C5 and 6179 had reached 9.1 log CFU/mL, while in dual culture, 6179 was affected by the presence of C5 attaining only 6.4 ± 0.8 log CFU/mL. FTIR-ATR spectra of CFSM produced by singly-cultured 6179 and the co-culture were almost identical. Characteristic peaks in FTIR-ATR spectrum of CFSM of singly-cultured C5 at 1741, 1645 and 1223 cm−1 represent functional groups which were not present in the CFSM of the co-culture. These molecules may be located intracellularly or mounted on bacterial cell surface and removed from the supernatant during cell filtration of the co-culture. Both singly- and co-cultured 6179 managed to grow similarly regardless of CFSM origin. Contrarily, both singly- and co-cultured C5 managed to outgrow 6179 in CFSM which contained high concentration of C5 metabolites, while in CFSM produced by singly-cultured 6179, C5 did not grow, suggesting that the produced metabolites of strain 6179 appears to be harmful to strain C5. However, during co-culture, C5 may produce molecules that counteract the inhibitory effect of 6179. The findings shed more light on the mechanism behind the inter-strain interactions of L. monocytogenes indicating that both contact of cells and extracellular metabolites may influence the behavior of the different co-existing strains.