Phytopathogenic organisms and mycotoxigenic fungi: Why do we control one and neglect the other? A biological control perspective in Malaysia

In this review, we present the current information on development and applications of biological control against phytopathogenic organisms as well as mycotoxigenic fungi in Malaysia as part of the integrated pest management (IPM) programs in a collective effort to achieve food security. Although the biological control of phytopathogenic organisms of economically important crops is well established and widely practiced in Malaysia with considerable success, the same cannot be said for mycotoxigenic fungi. This is surprising because the year round hot and humid Malaysian tropical climate is very conducive for the colonization of mycotoxigenic fungi and the potential contamination with mycotoxins. This suggests that less focus has been made on the control of mycotoxigenic species in the genera Aspergillus, Fusarium, and Penicillium in Malaysia, despite the food security and health implications of exposure to the mycotoxins produced by these species. At present, there is limited research in Malaysia related to biological control of the key mycotoxins, especially aflatoxins, Fusarium‐related mycotoxins, and ochratoxin A, in key food and feed chains. The expected threats of climate change, its impacts on both plant physiology and the proliferation of mycotoxigenic fungi, and the contamination of food and feed commodities with mycotoxins, including the discovery of masked mycotoxins, will pose significant new global challenges that will impact on mycotoxin management strategies in food and feed crops worldwide. Future research, especially in Malaysia, should urgently focus on these challenges to develop IPM strategies that include biological control for minimizing mycotoxins in economically important food and feed chains for the benefit of ensuring food safety and food security under climate change scenarios.     

Recent developments and applications of hyperspectral imaging for rapid detection of mycotoxins and mycotoxigenic fungi in food products

Mycotoxins are the foremost naturally occurring contaminants of food products such as corn, peanuts, tree nuts, and wheat. As the secondary metabolites, mycotoxins are mainly synthesized by many species of the genera Aspergillus, Fusarium and Penicillium, and are considered highly toxic and carcinogenic to humans and animals. Most mycotoxins are detected and quantified by analytical chemistry-based methods. While mycotoxigenic fungi are usually identified and quantified by biological methods. However, these methods are time-consuming, laborious, costly, and inconsistent because of the variability of the grain-sampling process. It is desirable to develop rapid, non-destructive and efficient methods that objectively measure and evaluate mycotoxins and mycotoxigenic fungi in food. In recent years, some spectroscopy-based technologies such as hyperspectral imaging (HSI), Raman spectroscopy, and Fourier transform infrared spectroscopy have been extensively investigated for their potential use as tools for the detection, classification, and sorting of mycotoxins and toxigenic fungal contaminants in food. HSI integrates both spatial and spectral information for every pixel in an image, making it suitable for rapid detection of large quantities of samples and more heterogeneous samples and for in-line sorting in the food industry. In order to track the latest research developments in HSI, this paper gives a brief overview of the theories and fundamentals behind the technology and discusses its applications in the field of rapid detection and sorting of mycotoxins and toxigenic fungi in food products. Additionally, advantages and disadvantages of HSI are compared, and its potential use in commercial applications is reported.     

Multiplex real-time PCR for detection and quantification of mycotoxigenic Aspergillus, Penicillium and Fusarium

The most agriculturally and economically important classes of mycotoxins are produced by species of Aspergillus, Penicillium, and Fusarium. Rapid methods to detect mycotoxigenic fungi could help prevent mycotoxins from entering the food chain. The purpose of this research was to develop a multiplex real-time PCR assay to detect and quantify multiple species of mycotoxigenic fungi. A pair of broad-spectrum PCR primers was designed for amplification of the internal transcribed spacer (ITS) regions of rDNA from the mycotoxigenic species. An in silico analysis of the primers revealed the presence of amplification in more than 40 Aspergillus species, 23 Fusarium species, and 32 Penicillium species as well as 64 other fungal genera. Genus-specific Taqman probes were designed from the ITS sequences of the most important mycotoxigenic species of Fusarium, Penicillium, and Aspergillus. The specificity of the probes was established against a wide range of fungal species. As a multiplex assay, the linear range of detection was 1 pg to 10 ng of DNA. The assay was validated by analyzing fungal growth in distiller's grain (DG), an animal feedstock that is a by-product when ethanol is produced from corn. This assay could be used as an initial step to evaluate the mycotoxigenic potential of DG and various other agricultural commodities.     

Exploration of Islamic medicine plant extracts as powerful antifungals for the prevention of mycotoxigenic Aspergilli growth in organic silage

BACKGROUND: Feed contamination with mycotoxins is a major risk factor for animals and humans as several toxins can exist as residues in meat and milk products, giving rise to carry‐over to consumers via ingestion of foods of animal origin. The starting point for prevention, in this chain, is to eliminate the growth of mycotoxigenic fungi in the animal forage. Ten plant extracts, recommended in Islamic medicine, were evaluated as antifungal agents against mycotoxigenic Aspergilli, i.e. Aspergillus flavus and A. ochraceus, growth in organic maize silage. RESULTS: Most extracts had remarkable antifungal activities using both qualitative and quantitative evaluation methods. Cress (Lepidium sativum) seed extract was proven to be the most powerful among the plants examined. Blending of the most effective extracts (garden cress seed, pomegranate peel and olive leaf extracts), individually at their minimal fungicidal concentrations, with maize silage resulted in the reduction of inoculated A. flavus colony counts by 99.9, 99.6 and 98.7%, respectively, whereas silage blending with the combined extracts completely prohibited fungal growth for up to 30 days of incubation under aerobic conditions. CONCLUSION: Besides the health promoting effects, silage blending with the bioactive plant extracts examined could lead to the required protection from pathogenic and mycotoxigenic fungi. Copyright © 2011 Society of Chemical Industry     

Mycotoxin Contamination of Rice in China

Mycotoxin contamination in rice is generally lower than in other cereals such as corn or wheat. However, over 65% of the population in China consumes rice as a staple food. Due to the diversity of the climate across China, the southern region is characterized by high temperatures and humidities, especially in rainy season. Such conditions are optimal for the growth of fungi. The accumulative and transferrable characteristics of fungi mycotoxins pose a great potential threat as confirmed by high incidences of liver cancer in the Yangtze delta region. Major mycotoxins identified in China are aflatoxins and ochratoxin A, as well as fumonisins. The contents of aflatoxin B₁ (AFB₁) in rice are varied among different provinces and regions and generally less than 5 μg/kg. Although high incidences of positive aflatoxins samples have widely been detected, few samples were detected as exceeding the national's maximum residue limit (10 μg/kg). Limited information is available on risk assessment of human health hazards of mycotoxins in rice, children should be paid more attention to due to their having the highest mycotoxins exposure level, although the risks are generally at low levels from rice. Mycotoxins are mainly distributed in the outer layer of the paddy rice (also called rough rice, referring to whole rice grain with the hulls), and the AFB₁ content in bran is 8.4 times greater than that in brown rice (hulled rice). Further investigation should focus on isolation and identification of mycotoxins‐producing fungal strains, especially unknown mycotoxigenic fungal strains determination. Infection resistant rice breeding of mycotoxigenic fungal species may be a fundamental approach to guaranteeing rice safety in China.     

Contamination of freshly harvested Bambara groundnut (Vigna subterranea) seed from Mpumalanga,South Africa,with mycotoxigenic fungi

Freshly harvested Bambara groundnut (BGN) is occasionally consumed raw and can potentially become infected with mycotoxingenic field fungi. In this study, BGN samples were obtained from 12 farms in three districts of Mpumalanga in South Africa. Eight pooled samples were screened for multi-mycotoxin contamination using Ultra Performance Liquid-Chromatography tandem mass spectrometry (UPLC-MS/MS). To identify mycoflora, 12 samples were screened using conventional and molecular methods. Selected potential mycotoxin producing isolates were screened for mycotoxins using UPLC-MS/MS. No mycotoxins were detected on the freshly harvested BGN samples, but they were infected with various mycotoxin producing fungal species namely Aspergillus flavus (50%), Penicillium citrinum (25%), Penicillium oxalicum (17%), Penicillium citreoviridin (0.8%), and Fusarium verticillioides (0.8%). Following screening of selected fungal cultures, aflatoxin B₁ (0.4, 0.45 and 0.4 ppm) and fumonisin B₁ (0.7 ppm) were detected from A. flavus and F. verticillioides, respectively. Identification of mycotoxigenic fungi on freshly harvested BGN presents a potential health risk.     

Biodiversity of complexes of mycotoxigenic fungal species associated with Fusarium ear rot of maize and Aspergillus rot of grape

Fusarium ear rot of maize and Aspergillus rot of grape are two examples of important plant diseases caused by complexes of species of mycotoxigenic fungi. These complexes of species tend to be closely related, produce different classes of mycotoxins, and can induce disease under different environmental conditions. The infection of maize and grape with multiple fungal species and the resulting production of large classes of mycotoxins is an example of mutual aggressiveness of microorganisms toward host species as well as to humans and animals that eat feed or food derived from the infected and contaminated plants. Infection of crop plant with a complex of microbial species certainly represents a greater threat to a crop plant and to human and animal health than infection of the plant with a single fungal species.     

Predicting mycotoxins in foods: A review

The need to ensure the microbiological quality and safety of food products has stimulated interest in the use of mathematical models for quantifying and predicting microbial behaviour. For 20 years, predictive microbiology has been developed for predicting the occurrence of food-borne pathogens, although these tools are dedicated to bacteria. Recently, the situation has changed and a growing number of studies are available in the literature dealing with the predictive modelling approach of fungi. To our knowledge the present one is the first review focussed on predictive mycology and food safety, including mycotoxins; existing kinetic and probability models applied to mycotoxigenic fungi germination and growth, and mycotoxin production are reviewed.     

Lactic Acid Bacteria as Antifungal and Anti‐Mycotoxigenic Agents: A Comprehensive Review

Fungal contamination of food and animal feed, especially by mycotoxigenic fungi, is not only a global food quality concern for food manufacturers, but it also poses serious health concerns because of the production of a variety of mycotoxins, some of which present considerable food safety challenges. In today's mega‐scale food and feed productions, which involve a number of processing steps and the use of a variety of ingredients, fungal contamination is regarded as unavoidable, even good manufacturing practices are followed. Chemical preservatives, to some extent, are successful in retarding microbial growth and achieving considerably longer shelf‐life. However, the increasing demand for clean label products requires manufacturers to find natural alternatives to replace chemically derived ingredients to guarantee the clean label. Lactic acid bacteria (LAB), with the status generally recognized as safe (GRAS), are apprehended as an apt choice to be used as natural preservatives in food and animal feed to control fungal growth and subsequent mycotoxin production. LAB species produce a vast spectrum of antifungal metabolites to inhibit fungal growth; and also have the capacity to adsorb, degrade, or detoxify fungal mycotoxins including ochratoxins, aflatoxins, and Fusarium toxins. The potential of many LAB species to circumvent spoilage associated with fungi has been exploited in a variety of human food and animal feed stuff. This review provides the most recent updates on the ability of LAB to serve as antifungal and anti‐mycotoxigenic agents. In addition, some recent trends of the use of LAB as biopreservative agents against fungal growth and mycotoxin production are highlighted.     

隐蔽型真菌毒素的形成及降解方法的研究进展

真菌毒素广泛分布于霉变的或受霉菌污染的粮食、谷物、饲料中,对人类和家畜健康造成严重威胁。植物和微生物可以降解真菌毒素为弱毒性的毒素,但是这种降解并不彻底,隐蔽型的真菌毒素依然存在于植物体内或环境中。本文主要从真菌毒素的分布及危害、植物生长和储存过程中降解真菌毒素的不完全性、微生物转化或降解真菌毒素形成“隐蔽型真菌毒素”、隐蔽型真菌毒素降解过程这四个方面进行论述,希望为今后真菌毒素的完全降解研究提供参考。     

Identification of fungal contamination and determination of mycotoxigenic molds by micellar electrokinetic capillary chromatography in smoked paprika

The purpose of this work was to analyze the fungal contamination in smoked and unsmoked paprika processed from different cultivars of pepper and to investigate the ability of these and other mycotoxigenic molds to grow and synthesize mycotoxins in smoked paprika. Eighteen mycotoxins were evaluated using micellar electrokinetic capillary chromatography. No relevant differences were found in fungal contamination between smoked and unsmoked paprika. The number of yeasts obtained was low, ranging from 0.4 to 3.29 log CFU g(-1); most of the yeast strains were identified as Cryptococcus spp. followed by Candida spp. All mold counts were <4 log CFU g(-1). Aspergillus, Cladosporium, Penicillium, and Fusarium were the predominant hyphomycete genera. Six mycotoxins were identified in the extracts of several strains isolated from paprika and incubated on malt extract agar. Penicillium expansum followed by Penicillium citrinum and Penicillium raistrickii were the dominant mycotoxigenic fungi isolated. Most of themycotoxin-producing fungi produced detectable amounts of mycotoxins when grown on paprika agar.     

Molecular biodiversity of mycotoxigenic fungi that threaten food safety

Fungal biodiversity is one of the most important contributors to the occurrence and severity of mycotoxin contamination of crop plants. Phenotypic and metabolic plasticity has enabled mycotoxigenic fungi to colonize a broad range of agriculturally important crops and to adapt to a range of environmental conditions. New mycotoxin-commodity combinations provide evidence for the ability of fungi to adapt to changing conditions and the emergence of genotypes that confer enhanced aggressiveness toward plants and/or altered mycotoxin production profiles. Perhaps the most important contributor to qualitative differences in mycotoxin production among fungi is variation in mycotoxin biosynthetic genes. Molecular genetic and biochemical analyses of toxigenic fungi have elucidated specific differences in biosynthetic genes that are responsible for intra- and inter-specific differences in mycotoxin production. For Aspergillus and Fusarium, the mycotoxigenic genera of greatest concern, variation in biosynthetic genes responsible for production of individual families of mycotoxins appears to be the result of evolutionary adaptation. Examples of such variation have been reported for: a) aflatoxin biosynthetic genes in Aspergillus flavus and Aspergillus parasiticus; b) trichothecene biosynthetic genes within and among Fusarium species; and c) fumonisin biosynthetic genes in Aspergillus and Fusarium species. Understanding the variation in these biosynthetic genes and the basis for variation in mycotoxin production is important for accurate assessment of the risks that fungi pose to food safety and for prevention of mycotoxin contamination of crops in the field and in storage.     

Integrated management of the risks of stored grain spoilage by seedborne fungi and contamination by storage mould mycotoxins – An update

Fungal spoilage of stored grains may occur when activity of water (aw) in cereal grain exceeds a critical limit enabling mould growth. Because it is not feasible to maintain all parts of large grain bulks below this critical moisture limit during prolonged storage time, an infection by seed-borne fungi is not rare in cereal grain stored under humid temperate or hot climates, inducing irreversible qualitative losses. Additionally, some fungal species produce harmful mycotoxins. The most harmful toxigenic species belong to the group of xerophilic species (genera Aspergillus and Penicillium). Because mycotoxin contamination of cereal grain is a worldwide issue for public health and a permanent concern for cereal-food industries facing the challenge of a permanent monitoring mycotoxin content in their primary matters, tolerable levels of mycotoxins are severely regulated worldwide. Mycotoxin-producing species growth is closely dependent of grain moisture levels enabling biological activity in grain ecosystem. Consequently, mould growth in stored grain bulks can be anticipated through early detection of grain and mould respiration. The prevention of mycotoxigenic fungi spoilage of stored grain can be managed by a preventive strategy. The main objective of the review was to describe the different methods, material and practices combined in such an integrated preventive approach. Some solutions potentially acceptable for the decontamination of moderately contaminated grain are also discussed.Integrated management of mould spoilage risks in stored grain is based on five pillars: i/Prevention of mould development by keeping grain moisture below the critical limit of fungal growth; ii/Accurate monitoring of grain aw and temperature changes during the storage period, associated to the monitoring of early indicators of respiration activity of storage fungi; iii/Reduction of grain bulk moistening trends by physical intervention means; iv/Use of physical treatments (ozone, grain peeling or abrasion) to limit mycotoxin contamination transfer to processed cereal products; v/Possible use of bio-competitive strains of fungi or bacteria to prevent the development of mycotoxigenic fungi in grain bulks. The future research needs on this topic are also evocated.     

Multiplex Real‐Time PCR Method for Detection and Quantification of Mycotoxigenic Fungi Belonging to Three Different Genera

Abstract: Cereal crop plants are colonized by many fungal species such as Aspergillus ochraceus and Penicillium verrucosum, which produce ochratoxins, and Fusarium graminearum, which produces trichothecene mycotoxins. A multiplex real‐time PCR method using TaqMan probes was developed to simultaneously detect and quantify these mycotoxigenic Fusarium, Penicillium and Aspergillus species in cereal grains. Primers and probes used in this method were designed targeting the trichothecene synthase (Tri5) gene in trichothecene‐producing Fusarium, rRNA gene in Penicillium verrucosum, and polyketide synthase gene (Pks) in Aspergillus ochraceus. The method was highly specific in detecting fungal species containing these genes and was sensitive, detecting up to 3 pg of genomic DNA. These PCR products were detectable over five orders of magnitude (3 pg to 30 ng of genomic DNA). The method was validated by evaluating sixteen barley culture samples for the presence of deoxynivalenol (DON) and ochratoxin A (OTA) producing fungi. Among the barley culture samples tested, 9 were positive for Fusarium spp, 5 tested positive for Penicillium spp, and 2 tested positive for Aspergillus spp. Results were confirmed by traditional microbiological methods. These results indicate that DON‐ and OTA‐producing fungi can be detected and quantified in a single reaction tube using this multiplex real‐time PCR method. Practical Application: This method would be helpful in detecting and quantifying the mycotoxin producing fungi such as Fusarium, Aspergillus, and Penicillium in cereal grains and cereal‐based foods.     

Occurrence of mycotoxigenic fungi and mycotoxins in animal feed from bihar,india

A total of 387 samples comprising animal feed, poultry feed and cattle cakes from India were examined in order to isolate mycotoxin-producing fungi as well as mycotoxins. Of 385 Aspergillus flavus group isolates 53% were capable of producing aflatoxins in liquid SMKY medium. Toxigenic strains of other mycotoxigenic fungi, viz A ochraceus, A versicolor, Penicillium citrinum and Fusarium moniliforme, were also recorded. In feed samples, beside the aflatoxins present in 139 samples zearalenone, ochratoxin A and citrinin were also recorded alone or as cocontaminants. A monsoon climate together with the socioeconomic conditions of this region are important determinants for the high incidence of mycotoxins in animal food.     

Advances in Mycotoxin Research: Public Health Perspectives

Aflatoxins, ochratoxins, fumonisins, deoxynivalenol, and zearalenone are of significant public health concern as they can cause serious adverse effects in different organs including the liver, kidney, and immune system in humans. These toxic secondary metabolites are produced by filamentous fungi mainly in the genus Aspergillus, Penicillium, and Fusarium. It is challenging to control the formation of mycotoxins due to the worldwide occurrence of these fungi in food and the environment. In addition to raw agricultural commodities, mycotoxins tend to remain in finished food products as they may not be destroyed by conventional processing techniques. Hence, much of our concern is directed to chronic health effects through long‐term exposure to one or multiple mycotoxins from contaminated foods. Ideally risk assessment requires a comprehensive data, including toxicological and epidemiological studies as well as surveillance and exposure assessment. Setting of regulatory limits for mycotoxins is considered necessary to protect human health from mycotoxin exposure. Although advances in analytical techniques provide basic yet critical tool in regulation as well as all aspects of scientific research, it has been acknowledged that different forms of mycotoxins such as analogs and conjugated mycotoxins may constitute a significant source of dietary exposure. Further studies should be warranted to correlate mycotoxin exposure and human health possibly via identification and validation of suitable biomarkers.     

A sub‐Saharan African perspective on mycotoxins in beer – a review

Beer is an alcoholic beverage made from a cereal grain extract and is widely consumed in sub‐Saharan Africa and the world at large. However, beer consumption could expose consumers to mycotoxins. In this review, we appraised the different mycotoxins associated with beer contamination, elucidating their structures and incidence in cereals involved in beer production. The common mycotoxins that are found within the brewing process are reviewed. These include aflatoxin B₁ (AFB₁), fumonisin (FB), ochratoxin A (OTA), zearalenone (ZEA) and deoxynivalenol (DON), which are the prime contaminants in beer produced in sub‐Saharan Africa. Residual levels of <20% of AFB₁, OTA and FB₂ together with the transformation of ZEA (into a less toxic compound β‐zearalenol) can be achieved during the production of beers originating from Europe/America, while >50% of DON and higher ratios of FB₁ can be recovered in finished beer. Adsorption is the major means of mycotoxin removal during beer production. In contrast, traditional African beer processes show no significant efficient removal of mycotoxins. This is because the prevailing environmental conditions during beer production are favourable to mycotoxigenic fungal proliferation. This subsequently leads to relatively high concentration of mycotoxins in freshly processed beer, with a possible increase during the beer shelf‐life owing to the absence of appropriate microbial stabilisation treatments in the finished processed beer. © 2019 The Institute of Brewing & Distilling     

Decontamination and detoxification strategies for the Fusarium mycotoxin deoxynivalenol in animal feed and the effectiveness of microbial biodegradation

Trichothecenes are a group of mycotoxins mainly produced by fungi of the Fusarium genus. Deoxynivalenol (DON) is one of the most abundant and important trichothecenes in food and feed, and is a significant contaminants due to its frequent occurrence in toxicologically relevant concentrations worldwide. Since toxin production depends strongly on environmental conditions, such as temperature and humidity, Fusarium toxin contamination can not be avoided completely. Therefore, exposure to this toxin is a permanent health risk for both humans and farm animals. As cereal crops are commonly contaminated with DON and animal diets consist mainly of cereals, it can be assumed that animals are frequently exposed to DON-contaminated feeds. Many strategies can be undertaken to reduce the toxic effect of DON. In addition to the general necessity for minimizing all risk factors that might influence the contamination of cereals with DON, such as the so-called field toxins before harvest, several post-harvest strategies can be applied to counteract possible deleterious effects of this mycotoxin in farm animals. Another approach for decontamination in feedstuffs is the use of adsorbent materials. Adsorbent materials may bind mycotoxins in the gastrointestinal tract and reduce absorption and systemic toxicity. It has been shown that some adsorbents are suitable to alleviate the toxic effects of specific mycotoxins, but its efficacy against trichothecenes is practically zero. Therefore, alternative strategies to reduce animal and human health risk are needed. The use of microbial additives is a method which uses microorganisms having the capability to detoxify mycotoxins by metabolism or degradation prior to their resorption in the gastrointestinal tract. DON has been reported to be completely transformed to de-epoxy-DON by ruminal and intestinal microflora. Eubacterium BBSH 797 was capable of DON degradation and counteracted the toxic effects of DON in animals. This review focuses on the efficacy of microbial feed additives in ameliorating the toxic effects of DON. According to the results of experiments to date, it appears that microorganisms are the main living organisms suitable for this mycotoxin biodegradation. However, the use of this approach depends on its effectiveness from both a practical and economic perspective.     

Mycoflora and natural occurrence of aflatoxin,cyclopiazonic acid,fumonisin and ochratoxin A in dried figs

Mycoflora, the mycotoxigenic properties of moulds, and natural contamination with mycotoxins such as aflatoxins (AFs), cyclopiazonic acid (CPA), fumonisin B₁ (FB₁) and ochratoxin A (OTA) were investigated in dried figs. Dry fig samples were collected from orchards during the drying stage in the Aegean Region of Turkey. Fungal isolates were identified using morphological, chemical as well as molecular methods. Mycotoxigenic characteristics of moulds were assessed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). Mycotoxins except CPA (by TLC) were determined by HPLC. All the fig samples were contaminated with moulds and 94.7% contained one or more mycotoxigenic species. The most prevalent moulds present in dried figs belong to the Aspergillus section Nigri members, being 93.9% positive for the samples, followed by Fusarium spp., Aspergillus section Flavi and Penicillium spp. On the other hand, Fusarium spp. had the highest count and the number of fumonisin producing Fusarium was also high. A total of 48% of 115 dried fig samples contained OTA (range?=?0.1–15.3?ng?g^?1), 74.7% of the samples had FB₁ (range?=?0.05–3.65?mg?kg^?1), 10.0% of the samples had aflatoxin (range?=?0.1–763.2?ng?g^?1) and 24.3% of the samples were tentatively identified as being contaminated with CPA (range?=?25–187?ng?g^?1). Dried fig samples were contaminated with one (33.0%), two (47.0%), three (5.2%) and four mycotoxins (3.5%). A total of 11.3% of dried fig samples were not contaminated with any of the four mycotoxins. To the best of our knowledge, CPA and fumonisin have been found for the first time in dried figs.     

Characterizing Soy Sauce Moromi Manufactured by High‐Salt Dilute‐State and Low‐Salt Solid‐State Fermentation Using Multiphase Analyzing Methods

Present study was to characterize the physiochemical properties, free amino acids (FAAs), volatiles and microbial communities of various moromi, respectively sampled from different stages of high‐salt dilute‐state (HSDS) and low‐salt solid‐state (LSSS) fermentation, using multiphase analyzing methods. Phospholipid fatty acids (PLFA) analysis indicated that Gram‐positive bacteria were dominant bacteria and fungi were principal microbes. For DGGE analysis, dominant microbes in moromi were mainly fell into Weissella, Tetragenococcus, Candida, Pichia, and Zygosaccharomyces. During fermentation, the dominant microbes shifted from nonhalophilic and less acid‐tolerant species to halophilic and acid‐tolerant species. Total of 15 FAAs and 44 volatiles were identified in moromi, mainly Glu, Asp, Tyr, and acids, alcohols, esters, aldehydes, respectively. Odor activity values analysis suggested that the final moromi of LSSS fermentation had more complicated odors than that of HSDS fermentation. Conclusively, technological parameters, microbial communities, raw materials and fermentation process may result in the discrepancy of HSDS and LSSS moromi.