Characterization of the complete mitochondrial genome of Paecilomyces variotii and comparative evolutionary mitochondriomics of 36 fungi.

BACKGROUD: Paecilomyces variotii has important economic value in stimulating crop growth, biodegradation, and other aspects. Up to now, there are no research reports on its mitochondrial genome. METHODS AND RESULTS: The mitochondrial genome of Paecilomyces variotii was determined with the next-generation sequencing method (Illumina, NovaSeq), and its characteristics were analyzed using various bioinformatics approaches. The length of complete mitochondrial genome sequence of P. variotii is 40,965 bp and consists of 14 protein-coding genes, 2 ribosomal RNA genes, 1 ribosomal protein S3 gene, 26 transport RNA genes. The results of phylogenetics analysis using Bayesian inference and Maximum likelihood methods showed that P. variotii belongs to the Eurotiales order in the Thermoascaceae family, and 9 genera within the Eurotiomycetes class were effectively distinguished with high support rates (bootstrap value > 92% and posterior probabilities > 99%). The analysis of synonymous substitution rates and nonsynonymous substitution rates indicated that the Ka/Ks values of the 14 PCGs in the mitochondrial genomes of the two orders in the Eurotiomycetes class ranged from 0 to 0.4333. CONCLUSIONS: This study revealed the structural and sequence information characteristics of the mitochondrial genome of P. variotii, and the phylogenetic results strongly support its classification within the family Thermoascaceae, consistent with traditional morphological taxonomy studies. The 14 PCGs in the mitochondrial genomes of the two orders in the Eurotiomycetes class are subject to strong purifying (negative) selection. The results of this research provides an important molecular basis for the development of genomics, evolutionary genetics and molecular markers of P. variotii in the future.

Integrated microbiome, metabolome and transcriptome profiling reveals the beneficial effects of fish oil and Bacillus subtilis jzxj-7 on mouse gut ecosystem.

The effects of fish oil (FO) and Bacillus subtilis jzxj-7 (JZXJ-7) on the colonic physiology, bacteria, metabolites, and gene expressions were studied in C57BL/6J mice. Co-administration of FO and JZXJ-7 was more beneficial than individual supplementation, as evidenced by improved growth performance, enhanced colon crypt depth and goblet cell numbers. FO + JZXJ-7 inhibited colonic fibrosis by downregulating fibrosis marker protein expression and upregulating occludin, claudin-2 and claudin-4 gene expressions. FO + JZXJ-7 ameliorated oxidative stress and inflammation by increasing catalase, superoxide dismutase, total anti-oxidation capacity, and reducing colon tumor necrosis factor (TNF)-α, interleukin (IL)-1ß and IL-6 levels. Mechanistically, FO + JZXJ-7 modulated the colon micro-ecological environment by enriching Roseburia, Lachnospiraceae NK4B4, Faecalibaculum and Lactococcus and its derived short-chain fatty acids, and activating Ppara and Car1 mediated peroxisome proliferators-activated receptor (PPAR) and phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt) signaling. Overall, FO + JZXJ-7 may serve as a promising nutraceutical to improve health by boosting the growth of colonic beneficial bacteria, altering metabolic phenotype, and regulating gene expression.

Effect of Amino Acids on Fusarium oxysporum Growth and Pathogenicity Regulated by TORC1-Tap42 Gene and Related Interaction Protein Analysis.

Free amino acids (AAs) formed in fermented meat products are important nitrogen sources for the survival and metabolism of contaminating fungi. These AAs are mainly regulated by the TORC1-Tap42 signaling pathway. Fusarium spp., a common contaminant of fermented products, is a potential threat to food safety. Therefore, there is an urgent need to clarify the effect of different AAs on Fusarium spp. growth and metabolism. This study investigated the effect of 18 AAs on Fusarium oxysporum (Fo17) growth, sporulation, T-2 toxin (T-2) synthesis and Tri5 expression through Tap42 gene regulation. Co-immunoprecipitation and Q Exactive LC-MS/MS methods were used to detect the interacting protein of Tap42 during specific AA treatment. Tap42 positively regulated L-His, L-Ile and L-Tyr absorption for Fo17 colony growth. Acidic (L-Asp, L-Glu) and sulfur-containing (L-Cys, L-Met) AAs significantly inhibited the Fo17 growth which was not regulated by Tap42. The L-Ile and L-Pro addition significantly activated the sporulation of ΔFoTap42. L-His and L-Ser inhibited the sporulation of ΔFoTap42. In T-2 synthesis, ΔFoTap42 was increased in GYM medium, but was markedly inhibited in L-Asp and L-Glu addition groups. Dose-response experiments showed that 10-70 mg/mL of neutral AA (L-Thr) and alkaline AA (L-His) significantly increased the T-2 production and Tri5 expression of Fo17, but Tri5 expression was not activated in ΔFoTap42. Inhibition of T-2 synthesis and Tri5 expression were observed in Fo17 following the addition of 30-70 mg/mL L-Asp. KEGG enrichment pathway analysis demonstrated that interacting proteins of Tap42 were from glycerophospholipid metabolism, pentose phosphate pathway, glyoxylate and dicarboxylate metabolism, glycolysis and gluconeogenesis, and were related to the MAPK and Hippo signaling pathways. This study enhanced our understanding of AA regulation in fermented foods and its effect on Fusarium growth and metabolism, and provided insight into potential ways to control fungal contamination in high-protein fermented foods.

Fish Oil Ameliorates Vibrio parahaemolyticus Infection in Mice by Restoring Colonic Microbiota, Metabolic Profiles, and Immune Homeostasis.

The effect of fish oil (FO) on colonic function, immunity, and microbiota was investigated in Vibrio parahaemolyticus (Vp)-infected C57BL/6J mice. Mice intragastrically presupplemented with FO (4.0 mg) significantly reduced Vp infection as evidenced by stabilizing body weight and reducing disease activity index score and immune organ ratios. FO minimized colonic pathological damage, strengthened the mucosal barrier, and sustained epithelial permeability by increasing epithelial crypt depth, goblet cell numbers, and tight junctions and inhibiting colonic collagen accumulation and fibrosis protein expression. Mechanistically, FO enhanced immunity by decreasing colonic CD3+ T cells, increasing CD4+ T cells, downregulating the TLR4 pathway, reducing interleukin-17 (IL-17) and tumor necrosis factor-α, and increasing immune cytokine IL-4 and interferon-γ levels. Additionally, FO maintained colonic microbiota eubiosis by improving microbial diversity and boosting Clostridium, Akkermansia, and Roseburia growth and their derived propionic acid and butyric acid levels. Collectively, FO alleviated Vp infection by enriching beneficial colonic microbiota and metabolites and restoring immune homeostasis.

Iturin A Strongly Inhibits the Growth and T-2 Toxin Synthesis of Fusarium oxysporum: A Morphological, Cellular, and Transcriptomics Study.

Fusarium oxysporum (F. oxysporum) is a common contaminant of dried fish, and the T-2 synthesis by this organism in dried fish products poses a serious public health risk. In this study, we investigated the effects of iturin A, a cyclic lipopeptide produced by Bacillus subtilis, on the growth and synthesis of the T-2 toxin of F. oxysporum, and transcriptomics was conducted. Results showed that the inhibitory effect of iturin A on F. oxysporum was significantly enhanced with an increase in iturin A concentrations. More specifically, compared with the control group, all indexes in the iturin A treatment group with 50 µg/mL were decreased to 24.84 mm, 0.33 × 106 cfu/mL, and 5.86 ng/mL for the colony diameter, number of spores, and concentration of T-2 toxin, respectively. Furthermore, iturin A was proven to destroy the integrity of cell membranes and cause a significant increase in ROS at 25 µg/mL or 50 µg/mL. Transcriptomic analysis revealed that with the treatment of iturin A, the genes of the oxidation-reduction process were up-regulated, while the gene expression of mycelial growth, cell integrity, transmembrane transport, energy metabolism, and others were down-regulated. More importantly, the Tri5 gene cluster was significantly inhibited. This study provided new insights into the mechanism for the inhibitory effect of iturin A on the growth and T-2 toxin synthesis of F. oxysporum and theoretical guidance for the application of iturin A in the preservation of dried aquatic products.

Cysteine Inhibits the Growth of Fusarium oxysporum and Promotes T-2 Toxin Synthesis through the Gtr/Tap42 Pathway.

Fusarium oxysporum is ubiquitous and can easily contaminate food during processing and storage, potentially producing T-2 toxin, which can pose a severe health risk to public health. Previous research on the presence of T-2 has focused on starch-rich foods, while protein- and amino acid-rich foods have received relatively little attention. In this study, the effects of amino acids on the growth of F. oxysporum and its T-2 production were investigated by gene deletion and complementation experiments. The results showed that amino acids, including aspartic acid, methionine, isoleucine, serine, phenylalanine, and cysteine, significantly inhibited the growth of F. oxysporum, while promoting T-2 synthesis, with cysteine having the most pronounced effect. The target of rapamycin complex 1 (TORC1) is a key pathway in response to a variety of amino acids, including cysteine. gtr2 and tap42 were found to be negative regulators of T-2 synthesis. The study highlights the elevated risk of T-2 production by F. oxysporum in cysteine-rich foods and the need to take appropriate measures to prevent and control the potential harm that such foods may present to public health. IMPORTANCE F. oxysporum and its T-2 contamination of food not only leads to food wastage but also poses a major food safety challenge to humans. The growth and T-2 production characteristics of F. oxysporum in high-protein substrates are considerably different from those in grains. Here, we show that the abundant free amino acids in a protein-rich food matrix are a key regulatory factor for the growth of, and toxin production by, F. oxysporum. Cysteine has the most pronounced effect on inhibiting mycelial growth and promoting T-2 synthesis through the TORC1 pathway. This implies that consumers tend to overlook T-2 contamination due to the poor growth of F. oxysporum in food rich in protein and amino acids, especially cysteine. Therefore, particular attention should be paid to the protection of those products.

Spatiotemporal Pattern Evolution and Driving Factors of Brucellosis in China, 2003-2019.

Brucellosis is a prevalent zoonotic disease worldwide. However, the spatiotemporal patterns evolution and its driving factors of Brucellosis have not been well explored. In this study, spatiotemporal scan statistics were applied to describe the spatiotemporal pattern of evolution in Brucellosis from 2003 to 2019 in mainland China, and GeoDetector analysis was further conducted to explore the driving effects of environmental, meteorological, and socioeconomic factors. We identified a distinct seasonal pattern for Brucellosis, with a peak in May and lowest incidence between September and December. High-risk clusters were first observed in the northwestern pastoral areas and later expanded to the southern urban areas. The spatiotemporal heterogeneity was mainly explained by total SO2 emissions, average annual temperature, sheep output, and consumption of meat per capita with explanatory powers of 45.38%, 44.60%, 40.76%, and 30.46% respectively. However, the explanatory power changed over time. Specifically, the explanatory power of average annual temperature tended to decrease over time, while consumption of meat per capita and total output of animal husbandry tended to increase. The most favorable conditions for the spread of Brucellosis include 0.66-0.70 million tons of SO2 emissions, 9.54-11.68 °C of average annual temperature, 63.28-72.40 million heads of sheep output, and 16.81-20.58 kg consumption of meat per capita. Brucellosis remains more prevalent in traditional pastoral areas in Northwest China, with the tendency of spreading from pastoral to non-pastoral, and rural to urban, areas. Total SO2 emission, average annual temperature, sheep output, and consumption of meat per capita dominated the spatial heterogeneity of Brucellosis with changes in explanatory power over time.

Comparative Analysis of Flavor, Taste, and Volatile Organic Compounds in Opossum Shrimp Paste during Long-Term Natural Fermentation Using E-Nose, E-Tongue, and HS-SPME-GC-MS.

The present study focused on the determination of color, flavor, taste, and volatile organic compounds (VOCs) changes of shrimp paste fermented for 1, 2, 3, and 8 years by E-nose, E-tongue, and headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS). During fermentation, the color of shrimp paste turned dark brown with decreases in L*, a*, and b* values. Inorganic sulfide odor was dominant in all fermented samples. The umami, richness, and aftertaste-B reached a maximum in year 3 of fermentation. A total of 182 volatiles, including long-chain alkanes, esters, aldehydes, olefins, ketones, acids, furans, and pyrazines, were detected. Sixteen VOCs including dimethyl disulfide, methional, trimethyl-pyrazine, (E,E)-2,4-heptadienal, benzeneacetaldehyde were selected as flavor markers. Correlation analysis showed that 94 VOCs were related to saltiness while 40, 17, 21, 22, and 24 VOCs contributed to richness, umami, aftertase-B, sourness, and bitterness, respectively. These novel data may help in optimizing fermentation duration to achieve target flavor indicators in opossum shrimp paste production.

Modulation of Intestinal Barrier, Inflammatory Response, and Gut Microbiota by Pediococcus pentosaceus zy-B Alleviates Vibrio parahaemolyticus Infection in C57BL/6J Mice.

Modulation of the intestinal barrier, inflammation, and gut microbiota by Pediococcus pentosaceus zy-B (zy-B) in Vibrio parahaemolyticus (Vp)-infected C57BL/6J mice was studied. Mice intragastrically pretreated with 108 colony-forming units (CFU) zy-B significantly alleviated Vp infection as evidenced by maintaining body weight and reduced disease activity index score and intestine ratio. In addition, zy-B reduced the Vp load in the ileum and cecum, significantly reduced the load in the colon, prevented colonic atrophy, and strengthened mucosal integrity. Mechanistically, zy-B ameliorated intestinal barrier dysfunction by upregulating tight junction protein expression, which in turn reduced the lipopolysaccharide, d-lactic acid (d-LA), and diamine oxidase concentrations and downregulated the cannabinoid receptor 1 (CB1) and CB2 mRNA expressions. Moreover, zy-B systemically reduced inflammation by decreasing interleukin-1ß (IL-1ß), interleukin-6 (IL-6), and tumor necrosis factor-α levels, and increased interleukin-10 (IL-10), immunoglobulin M (IgM), and immunoglobulin G (IgG) levels in the colon and serum. Furthermore, zy-B markedly altered the gut microbiota composition by enriching Bifidobacterium, Akkermansia, and Lactobacillus in the colon. Overall, zy-B appears to act as a probiotic to alleviate Vp infection by protecting the intestinal barrier, reducing inflammation, and promoting the growth of "beneficial" gut microbiota.

Effect of Food Matrix Type on Growth Characteristics of and Hemolysin Production by Vibrio alginolyticus.

ABSTRACT: The growth of and hemolysin production by two V. alginolyticus strains (HY9901 and ATCC 17749T) at 30°C were investigated in briny tilapia, shrimp, scallop, oyster, pork, chicken, freshwater fish, and egg fried rice. Bacteria were enumerated by plate counting. Hemolysin production was evaluated with blood agar and hemolytic titer tests. The two V. alginolyticus strains had similar growth and hemolysin production patterns in all tested foods. Based on the goodness-of-fit primary model statistics (coefficient of determination, mean square error, bias factor, and accuracy factor), the modified Gompertz model was a better fit than the logistic model to V. alginolyticus growth in foods. Growth kinetic parameters of V. alginolyticus had a higher µmax and shorter λ in the following order: briny tilapia > shrimp > freshwater fish > egg fried rice > scallop > oyster > chicken > pork. V. alginolyticus levels were similar at the stationary phase, with no significant growth difference between raw and cooked foods. Significantly higher thermostable direct hemolysin activity (P < 0.05) was found for V. alginolyticus in the following order: briny tilapia > freshwater fish > shrimp > chicken > egg fried rice > scallop > oyster > pork. However, the hemolytic titer was not consistent with the thermostable direct hemolysin activity and was significantly higher (P < 0.05) in the following order: briny tilapia > egg fried rice > shrimp > freshwater fish > chicken > scallop > oyster > pork. Contrary to current belief, V. alginolyticus produced more hemolysin in some nonseafoods (freshwater fish, egg fried rice, and chicken) than in scallops or oysters. This report is the first on the growth and toxicity of V. alginolyticus in different food matrices and confirms that some nonseafoods can be contaminated with pathogenic V. alginolyticus. These results should increase awareness of nonseafood safety issues and improve the accuracy of V. alginolyticus risk assessments.

Purification and Characterisation of Two Novel Pigment Proteins from the Carapace of Red Swamp Crayfish (Procambarus clarkii).

Pigment proteins play a vital role in the red colour change of the red swamp crayfish (Procambarus clarkii) shell after cooking. In this study, two red-change-related pigment proteins with molecular weights of approximately 170 and 43 kDa-denoted as F1 and F2, respectively-were purified by ammonium sulphate salting-out and size exclusion chromatography. F1 and F2 entirely comprised homomultimeric protein complexes composed of 21 kDa subunits. LC-MS/MS analysis showed that the 21 kDa protein subunit belonged to the crustacyanin family, named P. clarkii crustacyanin A2 (PcCRA2). The full-length cDNA of PcCRA2 was cloned, which encoded 190 amino acid residues and was highly homologous (91.58%) with Cherax quadricarinatus crustacyanin A. The predicted 3D structure showed that PcCRA2 had a ß-barrel structure for pigment encapsulation. The colour change of F1 was first detected at 40 °C, and the red change occurred upon heating above 60 °C. Additionally, with increasing temperature, its ß-sheet content increased, and its α-helix content reduced. Correlation analysis showed that the redness value of F1 was significantly related to the heating temperature and the ß-sheet content.

Simultaneous Quantification of Aflatoxin B1, T-2 Toxin, Ochratoxin A and Deoxynivalenol in Dried Seafood Products by LC-MS/MS.

Mycotoxins are secondary metabolites produced by fungi. These contaminate dried seafoods during processing and storage and represent a potential health hazard for consumers. A sensitive, selective and accurate liquid chromatography/tandem mass spectrometry (LC-MS/MS) method was established for simultaneous quantification of four common mycotoxins (aflatoxin B1 (AFB1), T-2 toxin (T-2), ochratoxin A (OTA) and deoxynivalenol (DON)) in dried shrimp, dried fish and dried mussel products. Mycotoxins were extracted from dried seafood samples by acetonitrile/water (85/15, v/v), subjected to ultrasound for 60 min at 20 °C and cleaned up by defatting with n-hexane. The sample matrix affected the linearity of detection (R2 ≥ 0.9974). The limit of detection (LOD) and limit of quantification (LOQ) in dried seafood products varied from 0.1 to 2.0 µg·kg-1 and 0.3 to 5.0 µg·kg-1, respectively. The method was validated by spiking samples with specific mycotoxin levels, and the recoveries, intra-relative standard deviation (RSDs) and inter-RSDs ranged between 72.2-98.4%, 2.8-10.6%, and 5.5-15.4%, respectively. This method was used to analyze 40 dried seafood products purchased from the Zhanjiang seafood market. Results of this product sampling showed that while no DON was detected, AFB1, T-2 and OTA were detected in 30.8%, 17.5% and 33.3% of the samples, respectively. AFB1, T-2 and OTA concentrations varied at 0.58-0.89, 0.55-1.34 and 0.36-1.51 µg·kg-1, respectively. Relatively high frequency of contamination and the presence of AFB1, OTA and T-2 residues indicate the need to monitor mycotoxins in dried seafood products.

Vibrio parahaemolyticus Infection in Mice Reduces Protective Gut Microbiota, Augmenting Disease Pathways.

Vibrio parahaemolyticus (Vp), a major food-borne pathogen, is responsible for severe infections such as gastroenteritis and septicemia, which may be accompanied by life-threatening complications. While studies have evaluated factors that affect the virulence of the pathogen, none have investigated the interaction of Vp with gut microbiota. To address this knowledge gap, we compared the effect of Vp on gut bacterial community structure, immunity, liver and kidney function, in pseudo germ-free (PGF) mice and normal (control) mice. Significant damage to the ileum was observed in normal mice compared with the PGF mice. The inflammatory factors IL-1ß, IL-6, and TNF-α in normal mice were ∼2.5-fold higher than in the PGF mice, and liver (ALT, AST, ALP) and kidney (BUN) function indices were ∼1.6-fold higher. The Vp infection substantially reduced species composition and richness of the gut microbial communities. In particular, there was a shift in keystone taxa, from protective species of genera Bacteroides, Lactobacillus, Bifidobacterium, and Akkermansia in the gut of control mice to opportunistic pathogens Enterobacteriaceae, Proteus, Prevotella, and Sutterella in Vp-infected mice, thus affecting microbiota-related biological functions in the mice. Specifically, pathways involved in infectious diseases and ion channels were significantly augmented in infected mice, while the pathways involved in metabolism, digestion and cell growth declined. We propose that the normal mice are more prone to Vp infection because of the alteration in gut-microbe-mediated functions. All these effects reduce intestinal resistance, with marked damage to the gut lining and pathogen leakage into the blood culminating in liver and kidney damage. These findings greatly advance our understanding of the mechanisms underlying interactions between Vp, the gut microbiota and the infected host.

Nanohybridization of Keggin polyoxometalate clusters and reduced graphene oxide for lithium-ion batteries.

The nanocomposites of reduced graphene oxide (RGO) and polyoxometalates (POMs) have been considered to be effective to boost more Li+ to participate in intercalation/deintercalation process of lithium-ion batteries (LIBs). In this paper, a nanocomposite (PMo12@RGO-AIL) with electrostatic interaction of RGO and Keggin-type [PMo12O40]3- has been fabricated and characterized by XRD, XPS, SEM, and TEM. To prepare PMo12@RGO-AIL, a strategy of covalent modification is developed between amino-based ionic liquid and RGO, helping to achieve the uniform dispersion of [PMo12O40]3-. When the PMo12@RGO-AIL was used as a cathode for LIBs, it could exhibit more excellent reversible capacity, cycle stability, and rate capability than those of samples without modifying by ionic liquids. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s11051-020-05108-x.

Diversity and succession of microbial communities and chemical analysis in dried Lutianus erythropterus during storage.

The population, diversity and succession of microbial communities and chemical characteristics of dried crimson snapper (Lutianus erythropterus) during storage for 50 days were investigated. The population of bacteria, yeasts and filamentous fungi in the samples were enumerated by culture methods using appropriate agar media. The amplicons of the 16S rRNA of bacteria and the ITS region of fungi were sequenced and compared with gene libraries to obtain the identity and abundance of microorganisms in the community. Free amino acids and several other chemical characteristics were determined by HPLC and corresponding chemical reaction methods. Before storage, the average counts of bacteria, yeasts and filamentous fungi in the dried fish were 3.2, 2.5 and 2.2 log CFU/g, which increased to 4.6, 3.6 and 3.9 log CFU/g, respectively after storage. Major succession in the bacterial > fungal communities occurred during storage as evidenced by a significant decline in the number and diversity of microbial communities. Predominant bacterial genera were Phytobacteria, Vibrio, Acinetobacter and Macrococcus in the freshly dried fish, which were replaced by Bacillaceae, Halomonas, Lentibacillus, Alkalibacillus after storage. The fungal community of the freshly dried fish consisted of Penicillum > Yamadazyma, Malassezia, Candida and Eurotiales with Penicillum being the most dominant. Penicillium camemberti was the most abundant fungal species in the dried fish before storage with most dominant after storage and accounted for >96% of the abundance. The succession in the microbial community was accompanied by major changes in chemical characteristics with a significant decrease in fat, and an increase in total free amino acids and total volatile basic nitrogen (TVB-N).

Effects of T-2 toxin on digestive enzyme activity, intestinal histopathology and growth in shrimp Litopenaeus vannamei.

T-2 toxin (T-2), a naturally occurring mycotoxin that often accumulates in aquatic animals via contaminated feed, is toxic to animals, including humans. In this study, six groups of shrimp (n = 30 shrimps/group) were given T-2 in feed at concentrations of 0-12.2 mg/kg for 20 days. T-2 accumulation, intestinal histopathology, digestive enzyme activities and subsequent effects on shrimp are reported. Compared to the control, T-2 significantly reduced weight gain, specific growth rate, and survival. The histopathology of shrimp intestine showed concentration-dependent degenerative and necrotic changes in response to dietary T-2. Progressive damage to the microstructures of shrimp intestine occurred with increasing dietary T-2 concentrations, with initial inflammation of the mucosal tissue at T-2 concentrations of 0.5 and 1.2 mg/kg, progressing to disappearance of intestinal villi and degeneration and necrosis of the submucosa at 12.2 mg/kg. Intestinal amylase and protease activities increased at low T-2 concentrations but showed significant inhibition at high concentrations; however, the opposite trend occurred for lipase activity. Collectively, these results indicate that digestive enzyme activities and mucosal structures are markedly affected by exposure to T-2, and these may have contributed to the lower survival rate of shrimp.

Influence of food matrix type on extracellular products of Vibrio parahaemolyticus.

BACKGROUND: Two strains of Vibrio parahaemolyticus (ATCC 17802 and 33847) in shrimp, oyster, freshwater fish, pork, chicken and egg fried rice were evaluated for production of hemolysin and exoenzymes of potential importance to the pathogenicity of this bacterium. RESULTS: The two strains of V. parahaemolyticus produced hemolysin, gelatinase, caseinase, phospholipase, urease, DNase and amylase in selected food matrices. Significantly higher (p < 0.05) hemolytic activity was produced by V. parahaemolyticus in egg fried rice > shrimp > freshwater fish > chicken > oyster > pork. But the exoenzyme activities were not consistent with the hemolytic activity profile, being significantly higher (p < 0.05) in shrimp > freshwater fish > chicken > oyster > pork > egg fried rice. Filtrates of V. parahaemolyticus from shrimp, freshwater fish and chicken given intraperitoneally to adult mice induced marked liver and kidney damage and were highly lethal compared with the filtrates of V. parahaemolyticus from oyster > egg fried rice > pork. CONCLUSION: From in vitro and in vivo tests, it appears that the food matrix type has a significant impact on the activity of extracellular products and the pathogenicity of V. parahaemolyticus. From a food safety aspect, it is important to determine which food matrices can stimulate V. parahaemolyticus to produce additional extracellular factors. This is the first report of non-seafood including freshwater fish and chicken contaminated with V. parahaemolyticus to have been shown to be toxic to mice in vivo.

Growth and Hemolysin Production Behavior of Vibrio parahaemolyticus in Different Food Matrices.

The growth and hemolytic activity profiles of two Vibrio parahaemolyticus strains (ATCC 17802 and ATCC 33847) in shrimp, oyster, freshwater fish, pork, chicken, and egg fried rice were investigated, and a prediction system for accurate microbial risk assessment was developed. The two V. parahaemolyticus strains displayed a similar growth and hemolysin production pattern in the foods at 37°C. Growth kinetic parameters showed that V. parahaemolyticus displayed higher maximum specific growth rate and shorter lag time values in shrimp > freshwater fish > egg fried rice> oyster > chicken > pork. Notably, there was a similar number of V. parahaemolyticus in all of these samples at the stationary phase. The hemolytic activity of V. parahaemolyticus in foods increased linearly with time ( R2 > 0.97). The rate constant ( K) of hemolytic activity was higher in shrimp, oyster, freshwater fish, and egg fried rice than in pork and chicken. Significantly higher hemolytic activity of V. parahaemolyticus was evident in egg fried rice > shrimp > freshwater fish > chicken > oyster > pork. The above-mentioned results indicate that V. parahaemolyticus could grow well regardless of the food type and that contrary to current belief, it displayed a higher hemolytic activity in some nonseafood products (freshwater fish, egg fried rice, and chicken) than in one seafood (oyster). The prediction system consisting of the growth model and hemolysin production algorithm reported here will fill a gap in predictive microbiology and improve significantly the accuracy of microbial risk assessment of V. parahaemolyticus.

Biotransformation enzyme activities and phase I metabolites analysis in Litopenaeus vannamei following intramuscular administration of T-2 toxin.

T-2 toxin (T-2) is a type-A trichothecene produced by Fusarium that causes toxicity to animals. T-2 contamination of grain-based aquatic feed is a concern for the industries related to edible aquatic crustacean species such as the shrimp industry because it can lead to serious food safety issues. T-2, its metabolites, and selected phase I (EROD, CarE) and phase II (GST, UGT, SULT) detoxification enzymes in hemolymph and tissues were monitored at 0, 5, 10 15, 30, 45, and 60 min following T-2 intramuscular administration (3 mg/kg bw) in shrimp (Litopenaeus vannamei). Marked increases of EROD activity in hepatopancreas and CarE activity in hemolymph, gill, hepatopancreas and intestine were observed followed by increases in phase II enzymes (GST, UGT, SULT) in hepatopancreas, hemolymph, intestine and gill, which remained elevated for an extended period. Time-dependent decrease in shrimp tissue T-2 concentration was observed. HT-2 increased up to 15 min. Most other T-2 metabolites were detected but not T-2 tetraol. Enzyme responses on exposure to T-2 were tissue-specific and time-dependent. Detection results indicated that HT-2 may not be the only important metabolite in aquatic crustacean species. Further investigation into T-2 metabolite toxicity is needed to fully understand the food safety issues related to T-2.

Effects of T-2 Toxin on Pacific White Shrimp Litopenaeus vannamei: Growth, and Antioxidant Defenses and Capacity and Histopathology in the Hepatopancreas.

Modified-masked T-2 toxin (mT-2) formed during metabolism in edible aquatic animals may go undetected by traditional analytical methods, thereby underestimating T-2 toxicity. The effects of T-2 on growth and antioxidant capacity and histopathological changes in the hepatopancreas were studied in Pacific white shrimp Litopenaeus vannamei exposed for 20 d to 0, 0.5, 1.2, 2.4, 4.8, and 12.2 mg/kg of T-2 in their feed. The concentration of mT-2 in the hepatopancreas was detected by liquid chromatography-tandem mass spectrophotometry before and after trifluoroacetic acid (TFA) treatment that converted mT-2 to free T-2. A dose-dependent increase in mT-2 concentration was observed in the hepatopancreas. Dietary exposure to T-2 significantly decreased (P < 0.05) shrimp growth and survival rate compared with the controls. The malondialdehyde (MDA) concentration was significantly increased in shrimp exposed to feed with ≥2.4 mg/kg T-2 (P < 0.05). The antioxidant enzymes, superoxide dismutase (SOD) and glutathione peroxidase (GPx), total antioxidant capacity (T-AOC), and also glutathione (GSH) content increased in shrimp dosed with 2.4-4.8 mg/kg T-2 but declined at the highest dose (12.2 mg/kg), probably indicating an inability to cope with high concentrations of reactive oxygen species (ROS) as evident from a marked increase in MDA (P < 0.05) culminating in cellular toxicity. Histopathological changes in the hepatopancreas were dose dependent, with cell autophagy evident at the highest exposure dose. This is the first report in shrimp of a dose-dependent increase in ROS, SOD enzyme activity, and T-AOC at low T-2 exposures, and associated histopathological changes in the hepatopancreas, in response to dietary T-2. Received January 26, 2016; accepted October 9, 2016.