Endophytic bacteria for Cd remediation in rice: Unraveling the Cd tolerance mechanisms of Cupriavidus metallidurans CML2.

The utility of endophytic bacteria in Cadmium (Cd) remediation has gained significant attention due to their ability to alleviate metal-induced stress and enhance plant growth. Here, we investigate C. metallidurans CML2, an endophytic bacterial strain prevalent in rice, showing resilience against 2400 mg/L of Cd(II). We conducted an in-depth integrated morphological and transcriptomic analysis illustrating the multifarious mechanisms CML2 employs to combat Cd, including the formation of biofilm and CdO nanoparticles, upregulation of genes involved in periplasmic immobilization, and the utilization of RND efflux pumps to extract excess Cd ions. Beyond Cd, CML2 exhibited robust tolerance to an array of heavy metals, including Mn2+, Se4+, Ni2+, Cu2+, and Hg2+, demonstrating effective Cd(II) removal capacity. Furthermore, CML2 has exhibited plant growth-promoting properties through the production of indole-3-acetic acid (IAA) at 0.93 mg/L, soluble phosphorus compounds at 1.11 mg/L, and siderophores at 22.67%. Supportively, pot experiments indicated an increase in root lengths and a decrease in Cd bioaccumulation in rice seedlings inoculated with CML2, consequently reducing Cd translocation rates from 43% to 31%. These findings not only contribute to the understanding of Cd resistance mechanisms in C. metallidurans, but also underscore CML2's promising application in Cd remediation within rice farming ecosystems.

Biochemical characterization and molecular modification of a zearalenone hydrolyzing enzyme Zhd11D from Phialophora attinorum.

ZEN lactone hydrolase (ZHD) can hydrolyze zearalenone (ZEN) to less or non-toxic product, providing an environment-friendly way for food or feeds-containing ZENs detoxification. Here, a newly identified ZHD from Phialophora attinorum, annotated as Zhd11D, was characterized to exhibit highest activity against ZEN at pH 8.0 and 35 â„ƒ with a specific activity of 304.7 U/mg, which was far higher than most of the reported ZHDs. A nonspecific protein engineering method was introduced through fusing a segment of amphiphilic short peptide S1 at the N-terminus of Zhd11D, resulting in both improved activity (1.5-fold) and thermostability (2-fold at 40 â„ƒ). Biochemical analysis demonstrated that self-aggregation caused by intermolecular interactions between S1 contributed to the improvement of the enzymatic properties of Zhd11D. Additionally, S1-Zhd11D showed a higher hydrolysis rate of ZEN than Zhd11D in peanut oil.

Biocontrol efficacy of atoxigenic Aspergillus flavus strains against aflatoxin contamination in peanut field in Guangdong province, South China.

Application of atoxigenic strains of Aspergillus flavusto soils is the most successful aflatoxin biological control approach. The objective of this study was to evaluate the efficacies of native non-aflatoxin producing (atoxigenic) strains as a biocontrol agent in peanut field in China. The competitive atoxigenic A. flavus strains (JS4, SI1and SXN) isolated from different crops, in China were used for field evaluation. The strains applied during the growing season (June - October, 2016) in the field at rate of 25 kg inoculum/hectare. The colonization of these biocontrol agents has been investigated and the population of A. flavus communities in soil were determined. The incidences of toxin producing (toxigenic) A. flavus strains and aflatoxin contamination in peanuts were also determined. Treated plots produced significant reductions in the incidence of toxigenic isolates of A. flavus in soil. However, the total fungal densities were not significantly different (p > 0.05) after treatments. Large percentage of aflatoxin reductions, ranging from 82.8% (SXN) up to 87.2% (JS4) were recorded in treated plots. Generally, the results suggest that the strategy can be used to control aflatoxin contamination and continuous evaluation should be done.

Cu2+-modified hollow carbon nanospheres: an unusual nanozyme with enhanced peroxidase-like activity.

A Cu2+-modified carboxylated hollow carbon nanospheres (Cu2+-HCNSs-COOH) was designed with enhanced peroxidase-like activity for the detection of hydrogen peroxide (H2O2) and degradation of methylene blue (MB). Hollow polymer nanospheres were fabricated from aniline, pyrrole, Triton-100, and ammonium persulfate via confined interfacial copolymerization reaction, which can be pyrolyzed to create HCNSs with the hollow gap diameter of about 20 nm under high temperature. Combining the synergistic effect of coordination and electrostatic interaction, Cu2+-HCNSs-COOH was constructed by anchoring Cu2+ on the surface of HCNSs-COOH. Furthermore, Cu2+-HCNSs-COOH has higher affinity for 3,3',5,5'-tetramethylbenzidine and H2O2 of 0.20 mM and 0.88 mM, respectively. Based on the rapid response of Cu2+-HCNSs-COOH to H2O2, we constructed a colorimetric sensing platform by detecting the absorbance of the 3,3',5,5'-tetramethylbenzidine-H2O2 system at 652 nm for quantifying H2O2, which holds good linear relationship between 1 and 150 µM and has a detection limit of 0.61 µM. We also investigated the degradation of MB in the presence of Cu2+-HCNSs-COOH and H2O2, which can degrade 80.7% pollutants within 30 min. This research developed an unusual nanozyme for bioassays and water pollution treatment, which broadened the way for the rapid development of clinical diagnostics and water pollution treatment.

Is the rate of adverse reaction to rabies vaccine in China really as the public thinks?-based on a meta analysis.

OBJECTIVE: To evaluate the rate of adverse reaction toward rabies vaccine in China from 2008 to 2019, explore its characteristics and to provide a scientific and objective basis for future policy decisions. METHODS: Literature on the rate of adverse reaction to rabies vaccine in China from 2008 to 2019 was retrieved and collected in CNKI, Wanfang, VIP databases, PubMed and Embase. A meta analysis was carried out then. RESULTS: Totally, 35 articles were included. The combined rate of adverse reaction to rabies vaccine was 5.6% (95% CI = 5.1% - 6.0%). Adverse reactions to rabies vaccine were 11.3% and 4.5% before and after 2011, 5.3% and 7.1% in the eastern and midwestern regions, 13.2%, 25.5% and 3.7% in the tertiary hospitals, secondary hospitals and primary medical institutions, respectively. And 13.3% were in the areas where the Changchun immortal vaccine was used. The combined rate of adverse reaction to rabies vaccine inoculated in Changsheng vaccine area was 5.1%. CONCLUSION: After 2011, the rate of adverse reaction to rabies vaccine in China has decreased dramatically. The rate of adverse reaction to rabies vaccine in the midwestern regions is higher than the counterpart in the eastern regions. The primary medical institutions are lower than the counterpart in the tertiary and secondary hospitals.

Prediction of Delayed Neurodevelopment in Infants Using Brainstem Auditory Evoked Potentials and the Bayley II Scales.

Background: Brainstem auditory evoked potentials (BAEP) provide an objective analysis of central nervous system function and development in infants. This study proposed to examine the relationship between infant BAEP values at age 6 months, and their neurodevelopment at age 2 years assessed by the mental development indices (MDI), a form of Bayley Scales of Infant Development. We hypothesized that in infants with BAEP values outside normal range, there may be neurodevelopmental delays, as shown by their MDI scores. Methods: An exploratory investigation was conducted using preterm (28-36 weeks gestation; 95 cases) and term infants (≥37 weeks gestation; 100 cases) who were born with specific perinatal conditions. BAEP values were recorded in these infants from 1 to 8 months of age, and compared with MDI scores in these infants at age 2 years. A multivariate linear regressions model was performed to test the associations between all variables and MDI scores. Stratified linear regression was used to test the interactions between gestational age and BAEP values with MDI scores. Significance was determined at a p < 0.05. Results: We found that BAEP values were inversely associated with MDI scores in premature infants (ß = -1.89; 95% confidence interval = -3.42 to -0.36), and that the effect of gestational age and BAEP values on the MDI scores is decreased by 1.89 points due to the interaction between these two variables. In premature babies, the lower the BAEP value below the mean, the greater the decrease in MDI score at age 2 years. Asphyxia and lower socioeconomic status in the family were also covariates associated with lower MDI scores at age 2 years. Conclusion: The data provided evidence that BAEP values outside the normal range in premature infants at age 6 months may predict developmental delays in cognitive and motor skills, as shown by MDI scores. We propose that BAEP assessment may be utilized as a potential indicator for neurodevelopment, and suggest that early intellectual and public health interventions should be encouraged to enrich neurodevelopment in premature babies with BAEP values outside the normal range.

Embodied Emotion Regulation: The Influence of Implicit Emotional Compatibility on Creative Thinking.

The regulatory effect of embodied emotion on one's general emotion and the impact of the compatibility or incompatibility of the two types of emotion on creative thinking are still debatable. The purpose of this study is to investigate these issues experimentally. In Experiment 1, participants completed an explicit positive and negative emotion test [Positive and Negative Affect Schedule (PANAS)] and an implicit positive and negative emotion test [Implicit Positive and Negative Affect Test (IPANAT)] twice on a computer after emotional video priming was used to induce negative emotions and facial expression manipulation was performed to induce embodied positive or negative emotions. It was found that maintaining the expression of a suppressed smile was helpful in regulating negative emotions (p = 0.047). Specifically, the implicit negative emotions induced by facial expression manipulation had a positive regulating effect on the implicit negative emotions induced by the video (T1, M = 47.813; to T2, M = 44.188). In Experiment 2, the positive or negative emotions of the participants were induced using emotional videos, and facial expression manipulation was used to induce their embodied positive or negative emotions. Then, the participants completed a creative test by completing alternative use tasks (AUTs) and Chinese character riddles. The AUT fluency score in the emotionally compatible group was significantly higher than that in the emotionally incompatible group (p = 0.032), but while experiencing negative emotions, the emotionally compatible group had a significantly higher originality score and insight in Chinese character riddle score than the emotionally incompatible group (p = 0.017, p = 0.004). Therefore, embodied negative emotion has a significant regulating effect on implicit negative emotion. The compatibility of emotion activated by facial expression and viewing a video contributes to creative thinking, whereas the incompatibility of emotion hinders creative thinking. The compatibility of emotion under positive emotions improved thinking fluency, whereas under negative emotions, it activated originality and insight in creative thinking. The influence of such emotional compatibility on creative thinking may be due to the regulating effect of embodied emotions on implicit emotions induced by emotional stimuli.

Hollow copper sulfide nanocubes as multifunctional nanozymes for colorimetric detection of dopamine and electrochemical detection of glucose.

Nanozymes have fascinated increasing attention in the field of artificial enzyme. Designing an ideal nanozyme usually requires a synergic advantage of reasonable nanostructures and large specific surface area for ensuring excellent mimicking-enzyme catalytic activity. Here we report a CuS nanozyme with hollow nanocube structure (h-CuS NCs), which has a large surface area of 57.84 m2 g-1, and thus realizes excellent mimicking-enzyme catalytic activity. Expectedly, our directed design of h-CuS NCs nanozymes has an affinity for H2O2 of 0.94 mM, which is outstanding among the state-of-the-art Cu-based nanozymes. Furthermore, this nanozyme acts as a multifunctional catalyst to induce luminol chemiluminescence and oxide 3, 3', 5, 5'-tetramethylbenzidine (TMB) in the presence of H2O2, and displays distinguished electrocatalytic activity to glucose oxidation. More intriguingly, the nanozyme can produce a promising photothermal effect under the illumination of near-infrared light. This work will provide a prototype for rational design of distinct nanostructures as multifunctional nanozymes in the area of electrochemical sensing, mimicking-enzyme catalytic biosening and cancer therapy.

A new cold-active and alkaline pectate lyase from Antarctic bacterium with high catalytic efficiency.

Cold-active enzymes have become attractive biocatalysts in biotechnological applications for their ability to retain high catalytic activity below 30 °C, which allows energy reduction and cost saving. Here, a 1041 bp gene pel1 encoding a 34.7 KDa pectate lyase was cloned from a facultatively psychrophilic Antarctic bacterium Massilia eurypsychrophila and heterologously expressed in Escherichia coli. PEL1 presented the highest 66% identity to the reported mesophilic pectate lyase PLXc. The purified PEL1 exhibits the optimum temperature and pH of 30 °C and 10 toward polygalacturonic acid, respectively. PEL1 is a cold-active enzyme that can retain 60% and 25% relative activity at 10 °C and 0 °C, respectively, while it loses most of activity at 40 °C for 10 min. PEL1 has the highest specific activity (78.75 U mg-1) than all other reported cold-active pectinase, making it a better choice for use in industry. Based on the detailed sequence and structure comparison between PEL1 and PLXc and mutation analysis, more flexible structure and some loop regions may contribute to the cold activity and thermal instability of PEL1. Our investigations of the cold-active mechanism of PEL1 might guide the rational design of PEL1 and other related enzymes.

A Consensus Ochratoxin A Biosynthetic Pathway: Insights from the Genome Sequence of Aspergillus ochraceus and a Comparative Genomic Analysis.

Ochratoxin A (OTA) is a toxic secondary metabolite produced by Aspergillus and Penicillium species that widely contaminates food and feed. We sequenced and assembled the complete ∼37-Mb genome of Aspergillusochraceus fc-1, a well-known producer of OTA. Key genes of the OTA biosynthetic pathway were identified by comparative genomic analyses with five other sequenced OTA-producing fungi: A. carbonarius, A. niger, A. steynii, A. westerdijkiae, and Penicillium nordicum OTA production was completely inhibited in the deletion mutants (ΔotaA, ΔotaB, ΔotaC, ΔotaD, and ΔotaR1), and OTA biosynthesis was restored by feeding a postblock substrate to the corresponding mutant. The OTA biosynthetic pathway was unblocked in the ΔotaD mutant by the addition of heterologously expressed halogenase. OTA biosynthesis begins with a polyketide synthase (PKS), OtaA, utilizing acetyl coenzyme A (acetyl-CoA) and malonyl-CoA to synthesize 7-methylmellein, which is oxidized to OTß by cytochrome P450 monooxygenase (OtaC). OTß and l-ß-phenylalanine are combined by a nonribosomal peptide synthetase (NRPS), OtaB, to form an amide bond to synthesize OTB. Finally, OTB is chlorinated by a halogenase (OtaD) to OTA. The otaABCD genes were expressed at low levels in the ΔotaR1 mutant. A second regulator, otaR2, which is adjacent to the biosynthetic gene, could modulate only the expression of otaA, otaB, and otaD Thus, we have identified a consensus OTA biosynthetic pathway that can be used to prevent and control OTA synthesis and will help us understand the variation and production of the intermediate components in the biosynthetic pathway.IMPORTANCE Ochratoxin A (OTA) is a significant mycotoxin that contaminates cereal products, coffee, grapes, wine, cheese, and meat. OTA is nephrotoxic, carcinogenic, teratogenic, and immunotoxic. OTA contamination is a serious threat to food safety, endangers human health, and can cause huge economic losses. At present, >20 species of the genera Aspergillus and Penicillium are known to produce OTA. Here we demonstrate that a consensus OTA biosynthetic pathway exists in all OTA-producing fungi and is encoded by a gene cluster containing four highly conserved biosynthetic genes and a bZIP transcription factor.

Expression, functional analysis and mutation of a novel neutral zearalenone-degrading enzyme.

The crops and grains were often contaminated by high level of mycotoxin zearalenone (ZEN). In order to remove ZEN and keep food safe, ZEN-degrading or detoxifying enzymes are urgently needed. Here, a newly identified lactonohydrolase responsible for the detoxification of ZEN, annotated as Zhd518, was expressed and characterized. Zhd518 showed 65% amino acid identity with Zhd101, which was widely studied for its ZEN-degrading ability. A detailed activity measurement method of ZEN-degrading enzyme was provided. Biochemical analysis indicated that the purified recombinant Zhd518 from E. coli exhibited a high activity against ZEN (207.0 U/mg), with the optimal temperature and pH of 40 °C and 8.0, respectively. The Zhd518 can degrade ZEN derivatives, and the specific activities against α-Zearalenol, ß-Zearalenol, α-Zearalanol and ß-Zearalanol were 23.0 U/mg, 64.7 U/mg, 119.8 U/mg and 66.5 U/mg, respectively. The active sites of Zhd518 were predicted by structure modeling and determined by mutation analysis. A point mutant N156H exhibited 3.3-fold activity against α-Zearalenol comparing to Zhd518. Zhd518 is the first reported neutral and the second characterized ZEN-degrading enzyme, which provides a new and more excellent candidate for ZEN detoxifying in food and feed industry.

pH-Signaling Transcription Factor AopacC Regulates Ochratoxin A Biosynthesis in Aspergillus ochraceus.

In Aspergillus and Penicillium species, an essential pH-response transcription factor pacC is involved in growth, pathogenicity, and toxigenicity. To investigate the connection between ochratoxin A (OTA) biosynthesis and ambient pH, the AopacC in Aspergillus ochraceus was functionally characterized using a loss-of-function mutant. The mycelium growth was inhibited under pH 4.5 and 10.0, while the sporulation increased under alkaline condition. A reduction of mycelium growth and an elevation of sporulation was observed in Δ AopacC mutant. Compared to neutral condition, OTA contents were respectively reduced by 71.6 and 79.8% under acidic and alkaline conditions. The expression of AopacC increased with the elevated pH, and deleting AopacC dramatically decreased OTA production and biosynthetic genes Aopks expression. Additionally, the Δ AopacC mutant exhibited attenuated infection ability toward pear fruits. These results suggest that AopacC is an alkaline-induced regulator responsible for growth and OTA biosynthesis in A. ochraceus and this regulatory mechanism might be pH-dependent.

Isolation and characterization of Aspergillus flavus strains in China.

Important staple foods (peanuts, maize and rice) are susceptible to contamination by aflatoxin (AF)-producing fungi such as Aspergillus flavus. The objective of this study was to explore non-aflatoxin-producing (atoxigenic) A. flavus strains as biocontrol agents for the control of AFs. In the current study, a total of 724 A. flavus strains were isolated from different regions of China. Polyphasic approaches were utilized for species identification. Non-aflatoxin and non-cyclopiazonic acid (CPA)-producing strains were further screened for aflatoxin B1 (AFB1) biosynthesis pathway gene clusters using a PCR assay. Strains lacking an amplicon for the regulatory gene aflR were then analyzed for the presence of the other 28 biosynthetic genes. Only 229 (32%) of the A. flavus strains were found to be atoxigenic. Smaller (S) sclerotial phenotypes were dominant (51%) compared to large (L, 34%) and non-sclerotial (NS, 15%) phenotypes. Among the atoxigenic strains, 24 strains were PCR-negative for the fas-1 and aflJ genes. Sixteen (67%) atoxigenic A. flavus strains were PCRnegative for 10 or more of the biosynthetic genes. Altogether, 18 new PCR product patterns were observed, indicating great diversity in the AFB1 biosynthesis pathway. The current study demonstrates that many atoxigenic A. flavus strains can be isolated from different regions of China. In the future laboratory as well as field based studies are recommended to test these atoxigenic strains as biocontrol agents for aflatoxin contamination.

Genetic engineering modification and fermentation optimization for extracellular production of recombinant proteins using Escherichia coli.

As a common expression host, Escherichia coli has received more and more attention due to the recently developed secretory expression system, which offers advantages like reduced downstream bioprocesses and improved product quality. These advantages, coupled with high-density fermentation technology, make it a preferred system for large-scale production of many proteins utilized in industry and agriculture at a reduced process cost. To improve the secretion efficiency of target proteins, various strategies, including signal peptide optimization, periplasmic leakage, and chaperones co-expression have been developed. In addition, the optimization of the fermentation conditions such as temperature, inducer, and medium were also taken into account for the extracellular production in the high-density fermentation to reduce the cost of production. Here, these strategies ranging from genetic engineering to fermentation optimization were summarized for the future guidance of extracellular production of recombinant proteins using E. coli.

High copy and stable expression of the xylanase XynHB in Saccharomyces cerevisiae by rDNA-mediated integration.

Xylanase is a widely-used additive in baking industry for enhancing dough and bread quality. Several xylanases used in baking industry were expressed in different systems, but their expression in antibiotic free vector system is highly essential and safe. In the present study, an alternative rDNA-mediated technology was developed to increase the copy number of target gene by integrating it into Saccharomyces cerevisiae genome. A xylanase-encoding gene xynHB from Bacillus sp. was cloned into pHBM367H and integrated into S. cerevisiae genome through rDNA-mediated recombination. Exogenous XynHB expressed by recombinant S. cerevisiae strain A13 exhibited higher degradation activity towards xylan than other transformants. The real-time PCR analysis on A13 genome revealed the presence of 13.64 copies of xynHB gene. Though no antibiotics have been used, the genetic stability and the xylanase activity of xynHB remained stable up to 1,011 generations of cultivation. S. cerevisiae strain A13 expressing xylanase reduced the required kneading time and increased the height and diameter of the dough size, which would be safe and effective in baking industry as no antibiotics-resistance risk. The new effective rDNA-mediated technology without using antibiotics here provides a way to clone other food related industrial enzymes for applications.

Aflatoxin B1 inhibition in Aspergillus flavus by Aspergillus niger through down-regulating expression of major biosynthetic genes and AFB1 degradation by atoxigenic A. flavus.

Twenty Aspergillus niger strains were isolated from peanuts and 14 strains were able to completely inhibit AFB1 production with co-cultivation. By using a Spin-X centrifuge system, it was confirmed that there are some soluble signal molecules or antibiotics involved in the inhibition by A. niger, although they are absent during the initial 24h of A. flavus growth when it is sensitive to inhibition. In A. flavus, 19 of 20 aflatoxin biosynthetic genes were down-regulated by A. niger. Importantly, the expression of aflS was significantly down-regulated, resulting in a reduction of AflS/AflR ratio. The results suggest that A. niger could directly inhibit AFB1 biosynthesis through reducing the abundance of aflS to aflR mRNAs. Interestingly, atoxigenic A. flavus JZ2 and GZ15 effectively degrade AFB1. Two new metabolites were identified and the key toxic lactone and furofuran rings both were destroyed and hydrogenated, meaning that lactonase and reductase might be involved in the degradation process.

A Survey of Aflatoxin-Producing Aspergillus sp. from Peanut Field Soils in Four Agroecological Zones of China.

Peanut pods are easily infected by aflatoxin-producing Aspergillus sp.ecies from field soil. To assess the aflatoxin-producing Aspergillus sp. in different peanut field soils, 344 aflatoxin-producing Aspergillus strains were isolated from 600 soil samples of four agroecological zones in China (the Southeast coastal zone (SEC), the Yangtze River zone (YZR), the Yellow River zone (YR) and the Northeast zone (NE)). Nearly 94.2% (324/344) of strains were A. flavus and 5.8% (20/344) of strains were A. parasiticus. YZR had the highest population density of Aspergillus sp. and positive rate of aflatoxin production in isolated strains (1039.3 cfu·g-1, 80.7%), the second was SEC (191.5 cfu·g-1, 48.7%), the third was YR (26.5 cfu·g-1, 22.7%), and the last was NE (2.4 cfu·g-1, 6.6%). The highest risk of AFB1 contamination on peanut was in YZR which had the largest number of AFB1 producing isolates in 1g soil, followed by SEC and YR, and the lowest was NE. The potential risk of AFB1 contamination in peanuts can increase with increasing population density and a positive rate of aflatoxin-producing Aspergillus sp. in field soils, suggesting that reducing aflatoxigenic Aspergillus sp. in field soils could prevent AFB1 contamination in peanuts.

Novel Aflatoxin-Degrading Enzyme from Bacillus shackletonii L7.

Food and feed contamination by aflatoxin (AF)B1 has adverse economic and health consequences. AFB1 degradation by microorganisms or microbial enzymes provides a promising preventive measure. To this end, the present study tested 43 bacterial isolates collected from maize, rice, and soil samples for AFB1-reducing activity. The higher activity was detected in isolate L7, which was identified as Bacillus shackletonii. L7 reduced AFB1, AFB2, and AFM1 levels by 92.1%, 84.1%, and 90.4%, respectively, after 72 h at 37 °C. The L7 culture supernatant degraded more AFB1 than viable cells and cell extracts; and the degradation activity was reduced from 77.9% to 15.3% in the presence of proteinase K and sodium dodecyl sulphate. A thermostable enzyme purified from the boiled supernatant was designated as Bacillus aflatoxin-degrading enzyme (BADE). An overall 9.55-fold purification of BADE with a recovery of 39.92% and an activity of 3.85 × 10³ U·mg-1 was obtained using chromatography on DEAE-Sepharose. BADE had an estimated molecular mass of 22 kDa and exhibited the highest activity at 70 °C and pH 8.0, which was enhanced by Cu2+ and inhibited by Zn2+, Mn2+, Mg2+, and Li⁺. BADE is the major protein involved in AFB1 detoxification. This is the first report of a BADE isolated from B. shackletonii, which has potential applications in the detoxification of aflatoxins during food and feed processing.

Variation in fungal microbiome (mycobiome) and aflatoxins during simulated storage of in-shell peanuts and peanut kernels.

Internal transcribed spacer 2 (ITS2) sequencing was used to characterize the peanut mycobiome during 90 days storage at five conditions. The fungal diversity in in-shell peanuts was higher with 110 operational taxonomic units (OTUs) and 41 genera than peanut kernels (91 OTUs and 37 genera). This means that the micro-environment in shell is more suitable for maintaining fungal diversity. At 20-30 d, Rhizopus, Eurotium and Wallemia were predominant in in-shell peanuts. In peanut kernels, Rhizopus (>30%) and Eurotium (>20%) were predominant at 10-20 d and 30 d, respectively. The relative abundances of Rhizopus, Eurotium and Wallemia were higher than Aspergillus, because they were xerophilic and grew well on substrates with low water activity (aw). During growth, they released metabolic water, thereby favoring the growth of Aspergillus. Therefore, from 30 to 90 d, the relative abundance of Aspergillus increased while that of Rhizopus, Eurotium and Wallemia decreased. Principal Coordinate Analysis (PCoA) revealed that peanuts stored for 60-90 days and for 10-30 days clustered differently from each other. Due to low aw values (0.34-0.72) and low levels of A. flavus, nine of 51 samples were contaminated with aflatoxins.

Ochratoxin A Producing Fungi, Biosynthetic Pathway and Regulatory Mechanisms.

Ochratoxin A (OTA), mainly produced by Aspergillus and Penicillum species, is one of the most important mycotoxin contaminants in agricultural products. It is detrimental to human health because of its nephrotoxicity, hepatotoxicity, carcinogenicity, teratogenicity, and immunosuppression. OTA structurally consists of adihydrocoumarin moiety linked with l-phenylalanine via an amide bond. OTA biosynthesis has been putatively hypothesized, although several contradictions exist on some processes of the biosynthetic pathway. We discuss recent information on molecular studies of OTA biosynthesis despite insufficient genetic background in detail. Accordingly, genetic regulation has also been explored with regard to the interaction between the regulators and the environmental factors. In this review, we focus on three aspects of OTA: OTA-producing strains, OTA biosynthetic pathway and the regulation mechanisms of OTA production. This can pave the way to assist in protecting food and feed from OTA contamination by understanding OTA biosynthetic pathway and regulatory mechanisms.