Characteristic of Chocolate Candy Produced from Fermented Cocoa Bean with Lactobacillus plantarum HL-15 Starter Culture for Inhibition Growth of Mycotoxin-Producing Fungi

Chocolate candy is a snack product that is liked by many people.The presence of mycotoxin producing fungi is a problem in chocolate product.The objective of this research was to observe the characteristic of chocolate candy which is produced from fermented cocoa bean with Lactobacillus plantarum HL-15 starter culture.The cocoa bean(47.5 kg)is fermented by adding L.plantarum HL-15 culture about 500 mL(10^10 CFU/mL)in the new and old fermentation box and in another new and old fermentation box without adding culture.Chocolate candy processing is done based on standard processing of chocolate candy.The results showed that the addition of L.plantarum HL-15 culture in cocoa bean fermentation and use of the new fermentation box give lower fungi concentration on chocolate candy,reducing 1 log cycle from 1.7×10^3 to<10^2 colony/g,while the addition of L.plantarum HL-15 culture in the old fermentation box is not effect.The average aw value of chocolate candy is 0.64 and pH value is 6.7.Fat content of chocolate candy is about 44.9%-46.2%.

Establishment and Application of a Real-time PCR Method for Detecting stx2 Gene in Shiga Toxin-producing Escherichia coli(STEC)

[Objective] This study aimed to establish a real-time PCR method for de- tecting stx2 gene in Shiga toxin-producing E. coli （STEC）. [Method] According to the known STEC stx2 gene sequences published in GenBank, PCR primers and probes were designed based on the conserved region to construct recombinant plasmid as a positive template, thus optimizing the reaction conditions and establishing the real- time PCR method. [Result] A standard curve was established based on the opti- mized real-time PCR system, indicting a good linear correlation between the initial template concentration and Ct value, with the correlation coefficient F^e of above 0.995. The established method had a good specificity, without non-specific amplifica- tion for 10 non-STEC intestinal bacterial strains; the detection limit of initial template was 1.0x102 copies/μI, indicating a high sensitivity; furthermore, the coefficients of variation within and among batches were lower than 1% and 5% respectively, sug- gesting a good repeatability. [Conclusion] In this study, a real-time PCR method was successfully established for detecting STEC stx2 gene, which provided technical means for rapid detection of STEC in samples.

Morphological Characterization of Arbuscular Mycorrhizal Fungi Associated with the Rhizosphere According to the Age of Xanthosoma sagittifolium L. Schott Plants in the Field

The objective of this work was to carry out a morphological characterization of arbuscular mycorrhizal fungi in the rhizosphere of Xanthosoma sagittifolium L. Schott plants. The plant material used was the white and red cultivars of X. sagittifolium, belonging to age intervals of 3 - 6, 6 - 9, and 9 - 12 months. Three harvest sites were chosen in the Central Region of Cameroon. In each site, soil from the rhizosphere and plant roots was collected in a randomized manner. In the field, the agronomic parameters were evaluated. The physicochemical characteristics of the soils, the mycorrhization index, and the morphological characterization of the mycorrhizal types of each site were carried out. The results obtained show that the agronomic growth parameters varied significantly using the Student Newman and Keuls Test depending on the harvest sites. The soils’ pH in all sites was acidic and ranged between 4.6 and 5.8. The Nkometou site has a loamy texture while the Olembe and Soa sites have loam-clay-sandy and loam-clay textures respectively. The highest mycorrhization frequencies appeared at the Nkometou site, with 75 and 87.33% of the white and red cultivars plant roots at 6 - 9 and 3 - 6 months. The relative abundance of AMF arbuscular mycorrhizal fungal spores in the rhizosphere of X. sagittifolium plants varied with age and cultivar. There were 673 spores between 9 - 12 months in Nkometou in the red cultivar. Six AMF genera were identified in all the different soils collected: Acaulospora sp., Funneliformis sp., Gigaspora sp., Glomus sp., Scutellospora sp., and Septoglomus sp. The genus Glomus sp. was the most present at all age intervals in both cultivars.

Soil Physico-Chemical Properties and Different Altitudes Affect Arbuscular Mycorrhizal Fungi Abundance and Colonization in Cacao Plantations of Cameroon

This study aims to investigate the abundance of AMF according to soil properties and altitudes in different cacao plantations of Cameroon. Physico-chemical analyses were made on soil samples collected from three agro-ecological zones. Soil samples were also used to evaluate directly the AMF abundance following the various altitudes and after trapping by sorghum plant. The results showed that soil properties, AMF spore abundances and colonization fluctuated significantly at different altitudes. The most represented texture was sandy loam. The bimodal zone presented a homogeneous texture (sandy loam) in all its localities. Cacao soil chemical characteristics showed that, the highest nitrogen rate (0.47%;p 0.05, Scott-Knott test) was recorded at Melong in a monomodal zone while Tonga in the Western highlands displayed the lowest rate (0.13%). Soil P concentration was significantly high in monomodal zones (Mbanga and Melong). Soil pH level indicated that the soil from Tonga in the Western highlands was neutral (pH = 6.67), and soils of other localities under study were acidic with the lowest (4.75) pH level recorded at Melong in a monomodal zone. In soil samples, the highest spore density (1.03 spores/g soil) was observed at Ntui in Bimodal zone, while the lowest spore density (0.26 spores/g soil) was observed at Bafang in the Western highlands. Root colonization showed that the sample from Bokito in a bimodal zone displayed the best frequency of mycorrhization (86.11%) while the sample from Bafang in the Western highlands recorded the lowest (27.11%). The PCA analysis highlighted that available phosphorus, pH and altitude all strongly correlated with AMF root colonization ability and can be used as a predictor of AMF colonization ability in cacao rhizosphere.

Reclassification and Nomenclature of Common Pathogenic Fungi

The accurate identification of pathogenic fungi is crucial for diagnosing and treating fungal infections. Recent advancements in molecular biotechnology, phylogenetic analysis, and the requirements of the Melbourne Code have led to changes in the classification and naming of fungi. These changes have caused significant confusion for medical laboratories and clinical personnel. This article summarizes the classification and new names of common pathogenic fungi to enhance fungi identification skills and provide accurate information for clinical diagnosis and treatment.

Secondary Metabolites of Entomopathogenic Fungi, Biological Alternative for the Control of Agricultural Pests and Disease: Present and Perspectives

The use of entomopathogenic fungi (EF) in recent years has been highly effective against the different orders of insects considered pests of agricultural importance and their conidia have been commonly applied, but it has been reported that these are sensitive to the environmental conditions. For this reason, biopesticides products have been formulated based on secondary metabolites, recently. These biomolecules participate as biological control agent, such as: cyclic depsipeptides, amino acids, polyketides, polyphenols and terpenoids, affecting their morphology, life cycle and insect behavior. The use of secondary metabolites of entomopathogenic fungi opens the possibility of application in a more efficient way for the control of agricultural pests in a compatible with the environment and human health;therefore, it is important to know, analyzing the type of molecules, their effects, and their different methods of application.

Fungi Associated with Sand and Plants from Marine Coastlines: Potential Relevance for Human Health

The fungal community associated with beach sand and plants located along marine coasts are an under-studied area of research despite its potential relevance to human health. In this study, we isolated and identified the cultivable mycobiota associated with sand and plants collected along the coast of Gran Canaria (Spain) using culture-dependent and -independent methods. Clinically relevant species belonging to Cryptococcus spp. and related genera such as Naganishia and Papilotrema were isolated and identified from shoreline plants. Moreover, Candida tropicalis was isolated from beach sand, and Aspergillus fumigatus and Aspergillus terreus strains were associated with both types of samples (i.e., plants and beach sand). We conclude that beach sand and shoreline plants are potential reservoirs of fungi of high clinical interest. We recommend including beach sand and plants from the environment when assessing the quality of marine coastal systems. Our results open a framework for studying the natural marine environment and its role in the epidemiology of infectious diseases in order to more accurately manage public health.

Resistance of Microbial Community of Artemisia annua L.to Pathogenic Fungi

[Objectives]This paper was to figure out whether the dominant bacterial community has the role and effect of bacterial community and its defense mechanism against potential pathogenic fungi in Artemisia annua,and thus establish a systematic model of bacteria-fungus-plant.[Methods]Fifty-eight strains of bacteria and one strain of pathogenic fungi,Globisporangium ultimatum,were used for the experiments.These 58 bacterial strains were assembled into a bacterial community,and the bacteria with abundance in the top 1%were reassembled into a dominant bacterial community as measured by 16S rDNA.[Results]The growth of A.annua seedlings inoculated with bacterial communities and pathogenic fungi or dominant bacterial communities and pathogenic fungi was significantly better than that of A.annua seedlings inoculated with pathogenic fungi during in vitro confrontation,which was evident in both enzymatic and non-enzymatic antioxidant assays.[Conclusions]The results suggest that the dominant bacterial community has a crucial role as a representative core microbial community of synthetic bacterial community,which can protect plants by interfering with the growth of phytopathogenic fungi mediated by chemical signals,and can be used as the main synthetic community of biocides to achieve the effect of biocontrol.

Dynamic Study on Soil Fungi Population in Artificial Vegetation Areas of Shapotou in Ningxia

[Objective] The changes of fungi in different soil layers and sand dunes of artificial vegetation areas in Ningxia Shapotou during different times were studied. [Method]The number of soil fungi in different soil layers at same plot and in same soil layer at different plots were changed significantly.The quantitative distribution of fungi was the most in Cuiliugou but the fewest in quicksand among all soil types.[Result] The quantity of soil fungi in grasslands was varied in different soil layers at the same sample and in the same soil layer at different sample plots. The quantity of fungi from different soil types distributed mostly in Cuiliugou, a natural desert steppe, but little in quicksand. The species and quantity of fungi in CuiLiugou was the most in different types of sand-fixing lands. [Conclusion] The quantity of soil fungi in artificial vegetation areas raises with the increase of sand-fixing ages, The enhancement of vegetation construction and artificial management measures in inland is helpful for improving present situation of desertification.

A Recycling Agriculture Model Based on Edible Fungi Cultivation

[Objective]This study was to explore the model for the efficient utilization of agricultural resources,especially crop straw and livestock waste.[Methods]With Shandong Jiufa Edible Fungus Co.,Ltd.as a case,this paper described a recycling agriculture model based on edible fungi,and analyzed its economic,ecological and social benefit [Results]The recycling agriculture model,guided by the scale production,processing and vendition of medium for the culture edible fungi,mainly makes use of a large amount of agriculture wastes like wheat straw,livestock waste,for developing edible fungi industry,aiming at getting the intensive optimization of fund,technology,raw materials and products at village level and more surplus labors engaged in edible fungi production industry into reality.Moreover,this model provides multi-solutions to the utilization of a great deal of fungus dreg,which can not only solve the problem of fungus dreg generated in the edible fungi production,but also extend industrial chain and enhance the income of practitioners,further realizing maximum conversion of agricultural resources.[Conclusion]This model can make well use of agricultural resources and reduce the agricultural waste pollution,significantly increasing economic,ecological and social benefits.

Effect of Different Arbuscular Mycorrhizal Fungi Strains on Mineral Nutrition and Antioxidant Enzymes of Chrysanthemum morifolium

[Objective] The purpose was to study the effects of different arbuscular mycorrhizal fungi strains（AMF）on the mineral nutrition and antioxidant enzymes of Chrysanthemum morifolium.[Method] The pot experiment was conducted in the greenhouse,C.morifolium ＇Jinba＇ was inoculated with five kinds of AMF,N,P,K,malondial dehyde（MDA）content,as well as the superoxide dismutase（SOD）,peroxidase（POD） and catalase（CAT） activity in roots,leaves and petals of C.morifolium were measured at seedling and flowering stages.[Result] The G.i,G.e and G.m treatments could promote mineral nutrient absorption,increase N,P,K content in roots,leaves and petals of C.morifolium compared with the control without inoculation.The G.d、G.e and G.m treatments could significantly reduce MDA content in roots and petals,thus alleviating membrane permeability and lipid peroxidation.The G.i treatments could also improve the SOD,POD and CAT activities of C.morifolium,thereby increasing the capability of scavenging oxygen free radicals.[Conclusion] According to the comprehensive analysis,G.i was screened out as the best strain to improve mineral nutrition and antioxidant enzyme activities of C.morifolium.

Study on biodegradated ability of thirteen edible fungi to straw

The biodegradated abilities of 13 edible fungi to straw were studied. The results showed that all the experimental fungi except Tricholama mongolicum had definite biodegradated abilities to the lignin and cellulose of straw. The Ideal fungus for straw degradation was screened out as Pleurotus ostreatus, which showed a higher degradation ability for lignin (17.86%) and lower degradation rate for cellulose (2.24%), with a Selection Factor (SF) of 7.97. The degradation rates of lignin and cellulose for other fungi ranged from 2.30% to 16.54% and 5.60% to 17.32%, respectively, and the SF was very low in range of 0.14 to 2.24.The ratio of colony's diameters to the color-zone (d1/d2) and SF are negative correlation, with a correlation coefficient of -0.1476.

Study on the Biological Diversity of Nematode-trapping Fungi in Erhai Lake

[Objective]The research aimed to investigate the biological diversity of nematode-trapping fungi in the sediment of Erhai Lake.[Method]616 pieces of sediments were collected from Erhai Lake.The traditional classification and identification methods were used to isolate,purify and identify.[Result]3 genera and 22 species of nematode-trapping fungi were isolated.Arthrobotrys oligospora,A.musiformis and Dactylella leptospora were the dominant species,and their detection rates were 28.05%,16.04% and 8.92% respectively.By analyzing the diversity of nematode-trapping fungi in four seasons,it was found that the biological diversity was richer in summer,spring and autumn,and the diversity indexes were 2.59,2.47 and 2.34 respectively.The diversity index in winter was 1.48 and was lower.Species forming the adhesive nets were predominant;positive rate was 41.00%.[Conclusion]The rich nematode-trapping fungi resource existed in Erhai Lake,and its biological diversity had the seasonal variation characteristic.The nematode-trapping fungi which formed the viscous net were the dominant species in Erhai Lake.

Chemical Prevention of Myxomycetes Disease in Edible Fungi

[ Objective ] The paper was to find effective chemical methods to control myxomycetes disease in edible fungi. [ Method] Some low toxic and pollution- free chemical reagents were first selected to carry out indoor toxicity test, and their inhibition effects against myxomyeetes were measured. Then the reagents with small effect on the growth of Pleurotu.s ostreatua were selected according to the indoor bioassay results, and carried out control test against myxomyeetes disease in mushroom house. [ Results] The results showed that the combination of Ludanlan sephora flavescens 1 000 times liquid ＋ salicylic acid 300 times liquid, Clolrim- azole suppositories 1 000 times liquid, Ludanlan sophora flavescens 1 000 times liquid and salicylic acid 300 times liquid had the best control efficacy against the disease. They were the preferred reagents for the prevention of myxomycetes disease in production practice. [ Conclusion] The results provided basis for the effec- tive control d myxomycetes disease in edible fungi.

Effects of Material Physical Properties on White-rot Fungi Mycelial Growth

[ Objective] The research aimed to study effects of material physical properties on white-rot fungi mycelial growth and provide theoretical basis for further expanding the application range of white-rot fungi. [ Method Four common species of white-rot fungi were cultivated by wood meal fowl dung mixture in test tube and culture dishes. The relationship between physical properties of culture material and the growth of these mycelials were studied. [Result] The results showed the water retention capacity of culture material was decreased with the increasing of its grain size and porosity, but the decrease of its specific gravity reduced the material water retention. And the dehydration rate of medium prepared with these materials at the same moisture conditions tended to increase. These physical properties of material, such as grain size, specific gravity, porosity, water retention and water drainage, influenced the growth of white-rot fungi mycelial by affecting the moisture and ventilation condition of media. The results hinted that above material physical properties had feedback effects on the growth of white-rot fungi mycelia. [ Conclution] Physical properties of culture material have significant effects on the growth of white-rot fungi mycelial.

Effects of Arbuscular Mycorrhizal Fungi(AMF) on Growth of Upland Rice under Soil Pb Contamination

[Objective] This study aimed to investigate the effects of arbuscular mycor-rhizal fungi （AMF） on growth of upland rice under soil Pb contamination. [Method] Using potting method, the effects of Glomus mosseae on the growth of Oryzal sati-va L. under different soil Pb concentrations （0, 300, 600 mg/kg） were investigated. [Result] According to the results, the mycorrhizal colonization rate of upland rice in-oculated with Glomus mosseae was significantly reduced （P〈0.05） with the increase of Pb concentration in soil. Compared with non-inoculation treatment, inoculation of arbuscular mycorrhizal fungi significantly improved the biomass of upland rice and Pb concentration of upland rice roots with addition of 300 mg/kg Pb but significantly reduced Pb concentration of upland rice shoots, which was consistent with the re-duced R/S （P〈0.05）; with addition of 600 mg/kg Pb, inoculation of arbuscular mycor-rhizal fungi significantly improved the biomass of upland rice roots （P〈0.05） but sig-nificantly reduced Pb concentration of upland rice shoots and roots （P〈0.05）; there was no significant difference in R/S between inoculation treatment and non-inocula-tion treatment. [Conclusion] This study indicated that inoculating Glomus mosseae under certain Pb concentrations could to some extent al eviate the toxic effects of Pb on Oryzal sativa L.

Effects of Arbuscular Mycorrhizal Fungi on Upland Rice Oxidative Stress Induced by Cu and Pb Contamination in Soil

[Objective] This study aimed to investigate the effect of arbuscular mycor-rhizal fungi on upland rice oxidative stress induced by Cu and Pb contamination in soil. [Method] The upland rice seeds were sowed in pots, in which the soil was previously mixed with a certain amount of Glomus mosseae and 0, 100 and 200 mg/kg Cu, or 0, 300 and 600 mg/kg Pb. In the control treatment, Glomus mosseae was inactivated before mixed into the soil. Then, the physiological and chemical properties of the aboveground parts of rice plants were measured at mature stage. [Result] Compared with the control treatment （NM）, Glomus mosseae （GM） treat-ment inhibited the POD, CAT and SOD activity while increased the soluble protein content under 100 mg/kg Cu and 300 mg/kg Pb treatment, improved the POD and CAT activity and soluble protein content while decreased SOD activity under 200 mg/kg Cu. SOD and POD activity showed no significant difference between NM and GM treatment under 600 mg/kg Pb, but the CAT activity was enhanced and soluble protein content was decreased. [Conclusion] This study wil provide theoretical refer-ence for bioremediation of soil heavy metal pol ution.

Diversity of filamentous fungi in organic layers of two forests in Zijin Mountain

A study was conducted to evaluate the cultivable filamentous fungal diversity in organic layers (L, F, and H layers) and A1 layer of two main forest types, Pinus massoniana and Liguidambar formasana mixed forest and Quercus variabilis forest, in Zijin Mountain(325?N, 11848?E), Nanjing, China. A total of 67 taxa comprising 56 Deuteromycetes, 3 Zygomycetes, 5 Asco-mycetes and 3 unidentified fungi were recognized from samples from the forest floor of the two forest types. The most abundant group was Deuteromycetes. The dominant genera in both forests were Alternaria sp., Aspergillus sp., Cladosporium sp., Mucor sp., Penicillium sp., Rhizopus sp., Gliocladium sp. and Trichoderma spp. The fungal diversity was higher in the mixed forest than that in Q. variabilis forest. For both forest types, the maximum fungal diversity was found in layer F and there existed significantly different in fungal diversity between layer F and layer L. In the mixed forest, richness of fungi isolated from needle litter (P. massoniana) was lower than that from leaf litter (L. formasana). The richness of fungi from needle litter increased with the in-crease of forest floor depth, but for leaf litter, the fungal diversity decreased with the depth of forest floor. The co-species of fungi from the two forest types, as well as from two kinds of litters in mixed forest, increased with the depth of the forest floor. The succession of fungi along with the process of decomposition was discussed here. The results also showed that litter quality was a critical factor affecting fungal diversity.

Determination and Analysis of Heavy Metals Contents in Edible Fungi

[Objective] This study aimed to understand the status of heavy metal contamination in edible fungi, thereby providing some scientific basis for the safety of edible fungi. [Method] The heavy metal contamination status in four kinds of edible fungi, i.e., shitake mushroom, mushroom, agaric and needle mushroom, from Guiyang City and Tongren City, Guizhou Province was evaluated by flame atomic absorption spectrophotometry. [Result] Cu, Mn, Cr and Ni were detected in the four kinds of edible fungi from Guiyang and Tongren; the Cr contents in the four kinds of edible fungi from Guiyang and Tongren all exceeded the national standard, and the Cu contents all met the national standard; The Ni contents in agaric and needle mushroom from Guiyang, and the Ni contents in the four kinds of edible fungi from Tongren, all exceeded the national standard; the enrichment capacity of the four kinds of edible fungi for the four kinds of heavy metal elements ranked as Cu＇s Mn＇s Cr＇s Ni＇s; among the four kinds of edible fungi, mushroom showed the strongest enrichment capacity for the four kinds of heavy metal elements. [Conclusion] The heavy metal contamination in the four kinds of edible fungi was all severe in Guiyang and Tongren, especially in Tongren. There has been currently no national standard for Mn content in edible fungi, which needs to be developed as soon as possible.

Optimum conditions for pure culture of major ectomycorrhizal fungi obtained from Pinus sylvestris var. mongolica plantations in southeastern Keerqin sandy lands, China

The effects of medium, pH, water potential and temperature on the culture for three pure strains （Lactarius deliciosus, Boletus edulis and Lactarius insulsus） of ectomycorrhizal fungi from plantation forests of Mongolian pine （Pinus sylvestris var. mongolica） on sandy lands were observed to obtain the optimum conditions for the growth of ectomycorrhizal fungi. The results indicated that the three ectomycorrhizal fungi could grow well in the mediums containing natural components, such as vitamin, pine juice and yeast powder, pH had a slight effect on the growth of the three ectomycorrhizal fungi, and the optimum pH values were 6.0 for L. deliciosus, 5.0 for B. edulis, respectively. However, L. insulsus had a wide pH range, and it grew better than the other two strains in neutral and light alkalescent mediums. Water potential （produced by Polyethylene Glycol, PEG） had significant effects on the ecological adaptability for the tested three fungi strains. All of the three stains grow better at lower PEG concentration （100 g PEG.kg^-1 H2O）. The best water potential was 10% PEG concentration for all of the three stains. Temperatures, especially high temperatures induced the fungi death. The optimum temperature for the growth of ectomycorrhizal fungi was 25-28℃ for all of the three stains.