Hyphopichia Burtonii Research Articles

Effect of Ozonized Water against Pathogenic Bacteria and Filamentous Fungi on Stainless Steel

Ozone is a molecule that has gained increasing interest in recent years by food industries for sanitization of food-grade surfaces. Compared to chemical sanitizers such as chlorine, hydrogen peroxide, or peracetic acid, ozone shows undeniable advantages, such as the absence of by-products that should affect human health or the possibility of generating it when needed. Therefore, the aim of this paper was the assessment of the resistance to ozonized water of two pathogenic bacteria (Listeria monocytogenes, Salmonella) and of three airborne food-spoiling fungi (Aspergillus brasiliensis, Hyphopichia burtonii, and Penicillium nordicum) inoculated on stainless steel tiles and treated in static conditions with 1 to 6 mg L−1 (pathogens) or 8.5 mg L−1 (filamentous fungi). Ozonized water gave different results based on the tested microorganisms: pathogenic bacteria proved markedly more sensible to ozone than filamentous fungi, even if great differences were observed at inter- and intra-specific levels for both categories of microorganisms. Nevertheless, the non-linear inactivation kinetics of the studied strains made the calculation of a punctual F-value difficult, so in industrial practice, adequate tailoring of the treatments to be applied, based on the registered extrinsic factors and the industrial bio-burden, would be appropriate.

Cyberlindnera fabianii, an Uncommon Yeast Responsible for Gluten Bread Spoilage.

A single strain of yeast was isolated from industrial gluten bread (GB) purchased from a local supermarket. This strain is responsible for spoilage consisting of white powdery and filamentous colonies due to the fragmentation of hyphae into short lengths (dust-type spots), similar to the spoilage produced by chalk yeasts such as Hyphopichia burtonii, Wickerhamomyces anomalus and Saccharomycopsis fibuligera. The isolated strains were identified initially by traditional methods as Wickerhamomyces anomalus, but with genomic analysis, they were definitively identified as Cyberlindnera fabianii, a rare ascomycetous opportunistic yeast species with low virulence attributes, uncommonly implicated in bread spoilage. However, these results demonstrate that this strain is phenotypically similar to Wi. anomalus. Cy. fabianii grew in GB because of its physicochemical characteristics which included pH 5.34, Aw 0.97 and a moisture of about 50.36. This spoilage was also confirmed by the presence of various compounds typical of yeasts, derived from sugar fermentation and amino acid degradation. These compounds included alcohols (ethanol, 1-propanol, isobutyl alcohol, isoamyl alcohol and n-amyl alcohol), organic acids (acetic and pentanoic acids) and esters (Ethylacetate, n-propil acetate, Ethylbutirrate, Isoamylacetate and Ethylpentanoate), identified in higher concentrations in the spoiled samples than in the unspoiled samples. The concentration of acetic acid was lower only in the spoiled samples, but this effect may be due to the consumption of this compound to produce acetate esters, which predominate in the spoiled samples.

First record of Hyphopichia burtonii isolated from the storage pest Sitophilus zeamais and its bioactivity against mycotoxigenic fungi

Summary Corn weevil (Sitophilus zeamais) is one of the most destructive pests of corn seeds during storage. The weevil may be a vector of mycotoxigenic fungi or yeast contaminating seed lots. In this study, an unknown yeast species was isolated from corn weevils found in stored corn seeds. We hypothesized that this yeast had an antifungal activity thereby inhibiting growth of mycotoxigenic fungi in corn seeds. The yeast species was identified as Hyphopichia burtonii, using combined morphological and molecular assays, and its potential inhibitory activity was assessed in vitro (spread plate and dual culture) against three known mycotoxigenic fungi, Fusarium verticillioides, Aspergillus niger and A. fl avus. Screening of the antagonistic activity of the yeast isolate showed 50 – 69% colony growth inhibition of three fungi when the yeast was spread plated on PDA but only slight inhibition (5.8 – 13.7% growth inhibition) in the dual culture assay. The sporulation of the fungi was also affected at 57 – 96% and 29 – 40% in spread plating and dual culture assay, respectively. In addition, volatile and non-volatile fractions also showed a reduction in mycelial growth. Variable responses were observed among the mycotoxigenic fungi. Further research would be interesting on the potential utilization of the antagonistic yeast to reduce fungal growth and sporulation, and possible mitigation of mycotoxin contamination in corn grains. To our knowledge, this is the first record of H. burtonii isolated from an insect, specifically S. zeamais.

Sanitizing of stainless steel surfaces in the food industry: Effect of gaseous ozone against pathogens and filamentous fungi

AbstractThe aim of this work was to evaluate the resistance of two pathogenic bacteria (Listeria monocytogenes and Salmonella enterica subsp. enterica serotype Senftenberg), and three airborne food‐spoiling filamentous fungi (Hyphopichia burtonii, Penicillium nordicum, and Aspergillus brasiliensis ATCC 16404) to gaseous ozone on stainless steel. Tests were carried out by exposing inoculated tiles to gaseous ozone in a laboratorial chamber at concentrations up to 50 mg/L and at times up to 180 min (bacteria) or 300 min (filamentous fungi). For bacteria, the resistance to gaseous ozone of L. monocytogenes proved markedly higher than that of S. enterica. Their D‐values increased by seven times (from 27.3 to 200.7 min for the strain of L. monocytogenes studied; from 7.3 to 48.0 min for the Salmonella species studied) when the ozone concentration were reduced from 12 to 4 mg/L. For filamentous fungi, a substantial effect was observed only at 50 mg/L on single‐layered inocula: at these conditions, A. brasiliensis ATCC 16404 proved the most resistant strain to gaseous ozone, its D‐value (134.4 min) being higher than those registered for the tested strains of H. burtonii (9.9 min) and P. nordicum (17.1 min). The results demonstrated that the use of gaseous ozone as a sanitizing agent for environments and working surfaces seems to be a potential alternative to chemical sanitizers against the two pathogens studied, whereas it seemed effective against the three filamentous fungi studied only in limited circumstances.

Phyllosphere to Ferment: Site Conditions Structure Cacao Pod and Fermentation Microbiomes in Hawaiʻi

Microbial fermentation is crucial to the development of flavors in cacao-based products, but how the abiotic environment and pod surface microbiomes can contribute to these different sensory profiles is underdeveloped. To better understand the connections between environment, plant variety, associated pod phyllosphere microbiomes, and the fermentation microbiome, we characterized the fungi and bacteria on pod surfaces and early fermentations of two cacao varieties across wet and dry sites on the island of O‘ahu, Hawai‘i. We showed that site (and the environmental conditions linked to those sites) significantly influenced the microbiomes of both the pod phyllosphere and fermentations but did not significantly explain the differences between cacao varieties. Certain taxa of bacteria and fungi seemingly prefer the wet Kualoa Ranch site compared with the drier Urban Garden Center Site. Rare members of the pod phyllosphere communities became highly enriched during fermentation, where basidiomycetous yeasts ( Filobasidium sp., Hannaella zeae, Rhodotorula mucilaginosa, R. paludigena, and Sporobolomyces carnicolor) became dominant at the wet site, and both ascomycetous and basidiomycetous yeasts ( Hyphopichia burtonii, Meyerozyma sp., Yamadazyma phyllophila, Filobasidium sp., Kwoniella heveanensis, R. mucilaginosa, Sporobolomyces sp., and S. carnicolor) became dominant at the dry site. The agricultural production site where cacao plants are grown should be considered carefully because it is likely important in structuring microbes that are present on pod surfaces and the microbes (mostly yeasts) that will be carried forward into fermentation. These yeasts may contribute to the development of the unique fruity or floral flavors in chocolates produced in Hawai‘i. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

DNA metabarcoding analysis of fungal community on surface of four root herbs

ObjectiveAngelicae Sinensis Radix (ASR, Danggui in Chinese), Cistanches Herba (CH, Roucongrong in Chinese), Ginseng Radix et Rhizoma (PG, Renshen in Chinese), and Panacis Quinquefolii Radix (PQ, Xiyangshen in Chinese), widely used as medicine and dietary supplement around the world, are susceptible to fungal and mycotoxin contamination. In this study, we aim to analyze their fungal community by DNA metabarcoding. MethodsA total of 12 root samples were collected from three main production areas in China. The samples were divided into four groups based on herb species, including ASR, CH, PG, and PQ groups. The fungal community on the surface of four root groups was investigated through DNA metabarcoding via targeting the internal transcribed spacer 2 region (ITS2). ResultsAll the 12 samples were detected with fungal contamination. Rhizopus (13.04%−74.03%), Aspergillus (1.76%−23.92%), and Fusarium (0.26%−15.27%) were the predominant genera. Ten important fungi were identified at the species level, including two potential toxigenic fungi (Penicillium citrinum and P. oxalicum) and eight human pathogenic fungi (Alternaria infectoria, Candida sake, Hyphopichia burtonii, Malassezia globosa, M. restricta, Rhizopus arrhizus, Rhodotorula mucilaginosa, and Ochroconis tshawytschae). Fungal community in ASR and CH groups was significantly different from other groups, while fungal community in PG and PQ groups was relatively similar. ConclusionDNA metabarcoding revealed the fungal community in four important root herbs. This study provided an important reference for preventing root herbs against fungal and mycotoxin contamination.

Three new yeast species from flowers of Camellia sinensis var. assamica collected in Northern Thailand and their tannin tolerance characterization.

Our recent research study focused on Miang fermentation revealed that tannin-tolerant yeasts and bacteria play vital roles in the Miang production process. A high proportion of yeast species are associated with plants, insects, or both, and nectar is one of the unexplored sources of yeast biodiversity. Therefore, this study aimed to isolate and identify yeasts of tea flowers of Camellia sinensis var. assamica and to investigate their tannin tolerance, which is a property essential to Miang production processes. A total of 82 yeasts were recovered from a total of 53 flower samples in Northern Thailand. It was found that two and eight yeast strains were distinct from all other known species within the genera Metschnikowia and Wickerhamiella, respectively. These yeast strains were described as three new species, namely, Metschnikowia lannaensis, Wickerhamiella camelliae, and W. thailandensis. The identification of these species was based on phenotypic (morphological, biochemical, and physiological characteristics) and phylogenetic analyses of a combination of the internal transcribed spacer (ITS) regions and the D1/D2 domains of the large subunit (LSU) ribosomal RNA gene. The yeast diversity in tea flowers acquired from Chiang Mai, Lampang, and Nan provinces had a positive correlation with those acquired from Phayao, Chiang Rai, and Phrae, respectively. Wickerhamiella azyma, Candida leandrae, and W. thailandensis were the species uniquely found in tea flowers collected from Nan and Phrae, Chiang Mai, and Lampang provinces, respectively. Some of the tannin-tolerant and/or tannase-producing yeasts were associated with yeasts in the commercial Miang process and those found during Miang production, i.e., C. tropicalis, Hyphopichia burtonii, Meyerozyma caribbica, Pichia manshurica, C. orthopsilosis, Cyberlindnera fabianii, Hanseniaspora uvarum, and Wickerhamomyces anomalus. In conclusion, these studies suggest that floral nectar could support the formation of yeast communities that are beneficial for Miang production.

Making sugar-free bread with wheat flour and halotolerant <i>Hyphopichia burtonii</i>-enriched culture

We re-evaluated the feasibility of fermentation seeds made from Inaniwa udon, a traditional wheat noodle. Inaniwa udon dough was cultivated in a 5 % saline medium at 29 ˚C for three days (hereinafter ‘Hyphopichia-enriched starter’). Without adding sugar or yeast, this Hyphopichia-enriched starter could be used to make a primitive type of bread whose ingredients were limited to flour, salt and a starter. A typical strain of this culture, Hyphopichia burtonii M2, had much higher α-amylase activity than the type culture of H. burtonii, NBRC 10837T. When used for bread making, both strong and weak flour yielded bread with a specific volume of 2.0–2.2 cm3/g. The fresh bread crumb tended to be dark in color, with an L* of 63–70.

Screening of Yeasts Isolated from Baijiu Environments for Producing 3-Methylthio-1-propanol and Optimizing Production Conditions.

3-Methylthio-1-propanol (3-Met) is widely used as a flavoring substance and an essential aroma ingredient in many foods. Producing 3-Met by microbial transformation is green and eco-friendly. In the present study, one strain, YHM-G, which produced a high level of 3-Met, was isolated from the Baijiu-producing environment. Strain YHM-G was identified as Hyphopichia burtonii according to its morphological properties, physiological and biochemical characteristics, and ribosomal large subunit 26S rRNA gene D1/D2 domain sequence analysis. The optimal conditions for 3-Met production by YHM-G were obtained by single factor design, Plackett-Burman design, steepest ascent path design and response surface methodology as follows: 42.7 g/L glucose, pH 6, 0.9 g/L yeast extract, 6 g/L L-methionine (L-Met), culture temperature 28 °C, shaking speed 210 rpm, loading volume 50 mL/250 mL, inoculum size 0.5% (v/v), culturing period 48 h and 2.5 g/L Tween-80. Under these optimal conditions, the 3-Met production by strain YHM-G was 3.16 g/L, a value 88.1% higher than that before optimization. Strain YHM-G can also produce a variety of flavor compounds that are important for many foods. This strain thus has the potential to increase the abundance of 3-Met in some fermented foods and enhance their aroma profiles.

Potential of electrolyzed water to inactivate bread and cheese spoilage fungi

Controlling food spoilage fungi remains a challenge for food industries, and regulations on the usage of chemical disinfectants are becoming restrictive. Then, this study aimed to evaluate electrolyzed water (EW) as a sustainable alternative for food spoilage fungi inactivation. The experiment was carried out according to the protocol for testing the antifungal effects of chemical sanitizers by the European Committee for Standardization (CEN), using acidic electrolyzed water (AEW-AAC: 85 ppm; pH: 2.65; ORP: 1120 mV) and a basic electrolyzed water (BEW- pH: 11.12; ORP: −209 mV) to inactivate spoilage fungi strains from bread (Hyphopichia burtonii and Penicillium roqueforti) and cheese (P. roqueforti and Penicillium commune), besides the standard fungi for this type of essay (Candida albicans and Aspergillus brasiliensis). AEW presented a higher antifungal effect, inactivating an average of 89 % of the exposed population when compared to its respective BEW, which inactivates about 81.5 %. In general, the standard strains A. brasiliensis (ATCC 16404) and Candida albicans (ATCC 24433) were more sensitive to both AEW and BEW than the food-spoilage strains. Among those, P. roqueforti strains were the most sensitive, followed by P. commune strains, while H. burtonii strains were the most tolerant. EW can be a sustainable alternative for product surface and facility cleaning with further antifungal action when a sanitization step is not mandatory or needed. Future studies searching for conditions to improve the antifungal action of EW could make their industrial usage more viable.

A High-Throughput Absolute Abundance Quantification Method for the Characterisation of Daqu Core Fungal Communities

An inherent issue in high-throughput sequencing applications is that they provide compositional data for relative abundance. This often obscures the true biomass and potential functions of fungi in the community. Therefore, we presented a high-throughput absolute quantification (HAQ) method to quantitatively estimate the fungal abundance in Daqu. In this study, five internal standard plasmids (ISPs) were designed for the fungal ITS2 subregion with high length variations. Five ISPs were then utilised to establish standard curves with a quantitative concentration range of 103–107 cells/g, and this was used to quantify the core fungi, including Basidiomycota, Ascomycota, and Mucoromycota. Using three types of mature Daqu from different regions, we demonstrated that the HAQ method yielded community profiles substantially different from those derived using relative abundances. Then, the HAQ method was applied to the Daqu during fermentation. The initial formation of the Daqu surface occurred in the fourth stage, which was mainly driven by moisture. The key fungi that caused the initial formation of the Daqu surface included Hyphopichia burtonii, Saccharomycopsis fibuligera, and Pichia kudriavzevii. The initial formation of the Daqu core occurred in the fifth stage, which was mainly affected by moisture and reducing the sugar content. The key fungi that cause the initial formation of the Daqu core included S. fibuligera and Paecilomyces verrucosus. We conclude that the HAQ method, when applied to ITS2 gene fungal community profiling, is quantitative and that its use will greatly improve our understanding of the fungal ecosystem in Daqu.

Effectiveness of two UV-C light-emitting diodes (LED) systems in inactivating fungal conidia on polyethylene terephthalate

The present study aimed to assess the effects of two UV-C LED systems on three food-spoilage fungi (Hyphopichia burtonii, Penicillium brevicompactum, Aspergillus brasiliensis) that can contaminate food packaging materials. The UV-C LED systems were used at two distinct light intensities (9.0 mW/cm2 and 5.5 mW/cm2) for up to 800 s. Fungal conidia were deposited on polyethylene terephthalate lids as either a single (4–5 log CFU/lid) or a multiple layer of cells (5–6 log CFU/lid) to evaluate any difference due to a potential “shadow effect”. In the tests performed, A. brasiliensis was the most resistant fungus to the effects of UV-C LED irradiation, with its 1D-value varying from 23 to 58 s (9.0 mW/cm2) and from 46 to 58 s (5.5 mW/cm2) regardless of the light source used or the type of inoculum. In contrast, H. burtonii and P. brevicompactum were inactivated within 5 s when a single layer was used, whereas their 1D-values were equal to 2–4 s (H. burtonii) and to 3–13 s (P. brevicompactum) when a multilayer was tested. Given the low fungal contamination of food packaging devices and the possibility of developing tailored LED systems, UV-C LED technology was confirmed as an interesting method of sanitization for food packaging.Industrial relevance: This study provides data on the behaviour of two different UV-C LED systems in inactivating fungal conidia on polyethylene terephthalate surfaces. This study helps to understand how specific factors (e.g., spatial arrangement of cells; fluctuating fungal loads on packaging devices) can influence the UV-C LED system performance and therefore could be useful for both plant manufacturers and food industries to develop tailored UV-C LED systems.

Efficacy of weak acid preservatives on spoilage fungi of bakery products

Organic acids and their salts are usually the first choice in the bread industry to restrict fungal spoilage, but their efficacy is pH-dependent and spoilage by fungi remains as a common threat. Therefore, this study aimed to evaluate the susceptibility of spoilage fungi of bakery products to acetic, sorbic, and propionic acids at different pH. Penicillium roqueforti, Penicilium paneum, Aspergillus pseudoglaucus, Aspergillus montevidensis and Hyphopichia burtonii strains isolated from spoiled products had their minimum inhibitory concentration (MIC) defined by macrodilution. The concentrations tested were: (i) sorbic acid up to 32 mM; (ii) propionic acid up to 1024 mM and (iii) acetic acid up to 800 mM with pH adjusted in 4.5, 5.0, 5.0 and 6.0 after setting the agent concentration. The lowest MICs for all agents were obtained at pH 4.5, usually doubling with every 0.5 pH increase. P. roqueforti strains isolated from spoiled products were the most resistant to all tested preservatives; while strains of the related species P. paneum, showed similar tolerance to acetic and propionic acids but was double more susceptible to sorbic acid. Strains of A. pseudoglaucus and A. montevidensis were indistinctly susceptible to the preservatives and were the most susceptible species to propionic and acetic acids. H. burtonii strains demonstrated the most variable behaviour in comparison to the other strains being the most susceptible to sorbic acid, were like Aspergillus strains regarding propionic acid, but tolerate well acetic acid. Propionic acid concentrations usually allowed in baked goods are lower than the concentrations required to inhibit the most tolerant isolates tested in this study. The same is true for sorbic acid at higher pH levels. Spoilage species of bakery ware presents a distinct susceptibility profile to the preservatives commonly used in this sector, but the high tolerance observed is a cause of concern.

Beer production potentiality of some non-Saccharomyces yeast obtained from a traditional beer starter emao.

The recent realisation regarding the potentiality of the long-neglected non-Saccharomyces yeasts in improving the flavour profile and functionality of alcoholic beverages has pushed researchers to search for such potent strains in many sources. We studied the fungal diversity and the rice beer production capability of the fungal strains isolated from emao-a traditional rice beer starter culture of the Boro community. Fifty distinct colonies were picked from mixed-culture plates, of which ten representative morphotypes were selected for species identification, and simultaneous saccharification and beer fermentation (SSBF) assay. The representative isolates were identified as Hyphopichia burtonii (Hbur-FI38, Hbur-FI44, Hbur-FI47 & Hbur-FI68), Saccharomyces cerevisiae (Scer-FI51), Wickerhamomyces anomalus (Wano-FI52), Candida carpophila (Ccar-FI53), Mucor circinelloides (Mcir-FI60), and Saccharomycopsis malanga (Smal-FI77 and Smal-FI84). The non-Saccharomyces yeast strains Hbur-FI38, Hbur-FI44, Ccar-FI53, and Smal-FI77 showed SSBF capacity on rice substrate producing beer that contained 7-10% (v/v) ethanol. A scaled-up fermentation assay was performed to assess the strain-wise fermentation behaviour in large-scale production. The nutritional, functional, and sensory qualities of theSSBF strain fermented beer were compared to the beer produced by emao. All the strains produced beer with reduced alcohol and energy value while compared to the traditional starter emao. Beer produced by both the strains of H. burtonii stood out with higher ascorbic acid, phenol, and antioxidant property, and improved sensory profile in addition to reduced alcohol and energy value. Such SSBF strains are advantageous over the non-SSBF S. cerevisiae strains as the former can be used for direct beer production from rice substrates.

Amylolytic Fungi in the Ethnic Beer Starter “emao” and Their Beer-Producing Attributes

Naturally occurring autochthonous microbes associated with ethnic beer starters are diverse and important as they play different functional roles in beer fermentations. The study on culturable microbes from the ethnic rice beer starter “emao” of the Bodo community of Assam is limited. Here we isolated and identified the culturable fungal diversity associated with emao and screened them for beer-producing capability from glucose and starch substrates. Based on morphology and molecular characterization, the species identified were Candida glabrata (Cgla_RF2), Cyberlindnera fabianii (Cfab_RF37), Hyphopichia burtonii (Hbur_RF19), Mucor circinelloides (Mcir_RF48), Mucor indicus (Mind_RF25), Penicillium citrinum (Pcit_RF32), Rhodosporidiobolus ruineniae (Rrui_RF4 & Rrui_RF43), Saccharomyces cerevisiae (Scer_RF6), Saccharomycopsis fibuligera (Sfib_RF11), and Wickerhamomyces anomalus (Wano_RF3) among which the relative abundance (RA) of W. anomalus was the highest (24%) followed by C. glabrata and H. burtonii (16% in each). Five (Hbur_RF19, Sfib_RF11, Mind_RF25, Mcir_RF48, and Pcit_RF32) of eleven isolates showed amylase positive in the starch medium. Scer_RF6 showed the highest ethanol tolerance (14% v/v) followed by Hbur_RF19 (12% v/v), Cgla_RF2 (11% v/v) and Wano_RF3 (11% v/v). The amylase-positive strains produced beer-containing ethanol in the range of 3.17–7.3 (% v/v) from rice substrate. Although the rice beer produced by amylase-positive strains showed negligible pH difference, other parameters like ethanol, ascorbic acid, total phenol, and antioxidant properties were varied from beer to beer. Antibacterial activities shown by Mcir_RF48 and Pcit_RF32 against the test bacteria were higher with a 23–35 mm zone of inhibition than the other isolates. The present findings reveal the presence of fungi with antibacterial, amylolytic, ethanol fermenting, and antioxidant producing capacity in emao which could the source for future bioprospection.

Fungal diversity and mycotoxins detected in maize stored in silo-bags: a review.

Silo-bags are hermetic storage systems that inhibit fungal growth because of their atmosphere with low humidity, as well as low pH and O2 concentrations, and a high CO2 concentration. If a silo-bag with stored maize loses its hermetic nature, it favors the development of fungi and the production of mycotoxins. To the best of our knowledge, this is the first review on the diversity of fungal species and mycotoxins that were reported in maize stored under the environmental conditions provided by silo-bags. The genera Penicillium, Aspergillus and Fusarium were found more frequently, whereas Acremonium spp., Alternaria sp., Candida sp., Cladosporium sp., Debaryomyces spp., Epiconum sp., Eupenicillium spp., Eurotium sp., Eurotium amstelodami, Hyphopichia spp., Hyphopichia burtonii, Moniliella sp., Wallemia sp. and genera within the orden Mucorales were reported less recurrently. Despite finding a great fungal diversity, all of the studies focused their investigations on a small group of toxins: fumonisins (FBs), aflatoxins (AFs), deoxynivalenol (DON), zearalenone (ZEA), patulin (PAT), toxin T2 (T2) and ochratoxin (OT). Of the FBs, fumonisin B1 and fumonisin B2 presented higher incidence percentages, followed by fumonisin B3 . Of the AFs, the only one reported was aflatoxin B1. The mycotoxins DON, ZEA and OT were found with lower incidences, whereas PAT and T2 were not detected. Good management practices of the silo-bags are necessary to achieve a hermetically sealed environment, without exchange of gases and water with the external environment during the storage period. © 2022 Society of Chemical Industry.

Metagenomic analysis decodes the fungal diversity of bio-dynamic preparations

Aim: Biodynamic farming system involves use of 8 different biodynamic preparations (BD 500-BD 507). Multi functionality of any ecosystem is due to its microbial diversity and community composition of microbes. So the present study was aimed to determine the total fungal population viz. unculturable ones, metagenomic analysis was done. Methodology: In the present study, 18S rDNA sequencing of V3-V4 amplicon regions was performed to identify and characterize fungal diversity, which existed in these preparations. Results: Alpha diversity was found to be maximum in BD506 with 868 OTU (operational taxanomic units) and minimum in BD507 with 254 OTU. At phylum level, the most abundant phylum was Ascomycota as recorded in 7 BD preparations with exception in the BD 500 (Unassigned). At genus level highest percentage of OTU abundance was observed for unassigned genus in all BD preparations, except Mortierella in BD 500 and BD 502; Microascus in BD 501 and BD504; Gymnoascus in BD503, Scedosporium in BD 505, Mucor in BD 506 and Hyphopichia in BD 507. On the basis of species diversity, BD502, 503 and 506 showed high percentage of OTU abundance for Mucor racemosus, while Mortierella oligospora was abundant in BD500, Dipodascus geotrichum in BD 501, Kernia pachypleura in BD504, Petriella setifera in BD505 and Hyphopichia burtonii in BD 507. Interpretation: This indicated a unqiue class of fungus predominating each type of BD preparation. Furthermore, a large proportion of unassigned fungi at phylum and genus level were detected in metagenome analysis which might have specific roles in contributing for their overall effectiveness of each kind of BD preparations.

Integrated genomic and transcriptomic analysis reveals unique mechanisms for high osmotolerance and halotolerance in Hyphopichia yeast.

The yeast species Hyphopichia is common in nature and strongly competitive under harsh environmental conditions. Here, we characterized Hyphopichia burtonii KJJ43 and H. pseudoburtonii KJS14, which exhibit strong halotolerance, using genomic and transcriptomic analyses. The genomes of H. burtonii and H. pseudoburtonii comprised eight chromosomes with 85.17% nucleotide identity and significant divergence in synteny. Notably, both Hyphopichia genomes possessed extended gene families of amino acid permeases and ATP-binding cassette (ABC) transporters, whose dynamic expression patterns during osmotic stress were revealed using transcriptome profiling. Intriguingly, we found unique features of the HOG pathway activated by Hog1p even under non-osmotic stress conditions and the upregulation of cytosolic Gpd1 protein during osmotic stress. Associated with hyperfilamentation growth under high osmotic conditions, a set of genes in the FLO family with induced expression in response to NaCl, KCl, and sorbitol supplementation were identified. Moreover, comparative transcriptome analysis reveals the NaCl-specific induction of genes involved in amino acid biosynthesis and metabolism, particularly BAT2. This suggests the potential association between oxoacid reaction involving branched-chain amino acids and osmotolerance. The combined omics analysis of two Hyphopichia species provides insights into the novel mechanisms involved in salt and osmo-stress tolerance exploited by diverse eukaryotic organisms.

Comparative Growth Inhibition of Bread Spoilage Fungi by Different Preservative Concentrations Using a Rapid Turbidimetric Assay System.

Bread and intermediate moisture bakery products are mainly spoiled by yeasts and filamentous fungi. The inoculum load and preservation system used determines their shelf life. To extend the shelf life of such commodities, the use of chemical preservatives is the most common way to try and control the initiation of mold spoilage of bread. This study has utilized a rapid turbidimetric assay system (Bioscreen C) to examine the temporal efficacy of calcium propionate (CP) and potassium sorbate (PS) for controlling the growth of important bread spoilage fungi. The objectives were to compare the temporal growth of strains of three important spoilage fungi Hyphopichia burtonii (HB17), Paecilomyces variotii (PV11), and Penicillium roqueforti (PR06) isolated from visibly molded bread to (a) different concentrations of CP and PS (0–128 mM), (b) temperatures (25°C, 30°C), (c) water activity (aw; 0.95, 0.97), and (d) pH (5.0, 5.5). All three abiotic factors, pH, aw, and temperature, and preservative concentrations influenced the relative growth of the species examined. In general, PS was more effective than CP in inhibiting the growth of the strains of these three species. In addition, the Time to Detection (TTD) for the efficacy of the preservatives under the interacting abiotic factors was compared. The strain of Paecilomyces variotii (PV10) was the most tolerant to the preservatives, with the shortest TTD values for both preservatives. P. roqueforti was the most sensitive with the longest TTD values under all conditions examined. These results are discussed in the context of the evolution of resistance to food-grade preservatives by such spoilage fungi in bakery products.

Influence of environmental growth conditions on chalk yeasts causing bread spoilage

The main objective of this study was the investigation of different environmental growth conditions on the growth of chalk yeasts, Hyphopichia burtonii (HB), Wickerhamomyces anomalus (WA) and Saccharomycopsis fibuligera (SF). Through multiple methods, including radial growth, biocontrol assay, growth/no-growth modelling and a bread challenge test, the influence of pH (4.5–6.5), temperature (22–30 °C), propionic acid (0–0.3%) and presence of other spoilage fungi was assessed. This study showed that pH and temperature does not play a major role on the growth of HB, WA and SF. Only WA shows optimal growth at pH 4.5 and 30 °C on MEA. The likeliness of growth is the following: HB >SF >WA, except from WA at pH 4.5/30 °C. Biocontrol activity was observed against Aspergillus niger and Penicillium paneum in the presence of high concentrations of HB at 30 °C. Furthermore, this study showed that HB is most sensitive to propionic acid (PA), followed by SF and WA. The effectiveness of PA was also shown through bread challenge tests. The following order of sensitivity against PA was observed: HB >SF >WA. Breads inoculated with WA had a shorter shelf-life (2–3 days less).