Infección por Aspergillus flavus y Rhizopus oryzae complex en paciente con diabetes mellitus / Infection due to Aspergillus flavus and Rhizopus oryzae complex in a patient with diabetes mellitus

La sinusitis micótica es una condición patológica que puede presentarse en pacientes con diabetes mellitus y estar asociada a una crisis hiperglucémica. Es una entidad agresiva con complicaciones locales que incluyen afectación de la órbita y el sistema nervioso central, y compromiso vascular. A pesar del tratamiento quirúrgico y antimicótico, la mortalidad es de hasta el 75 %. Se describe el caso de una paciente con diagnóstico de cetoacidosis diabética y signos de oftalmoplejía unilateral que llevaron al estudio con resonancia magnética del sistema nervioso central; se encontraron signos de sinusitis, meningitis y cerebritis. Los estudios microbiológicos iniciales fueron negativos, y los biomarcadores galactomanano sérico y el antígeno de Cryptococcus también fueron negativos. Tras el manejo quirúrgico, se llegó a la identificación de Aspergillus flavus y Rhizopus spp. en el tejido de los senos paranasales. La paciente recibió tratamiento con posaconazol y, tras dos meses de seguimiento, había presentado mejoría clínica. La infección fúngica dual y la infección por A. flavus son entidades poco frecuentes y de relevancia clínica, sin casos presentados previamente en nuestro país por lo que este corresponde a un caso de interés clínico.

Fungal sinusitis is a pathology that can occur in patients with diabetes mellitus and be associated with a hyperglycemic crisis. It is an aggressive entity with local complications that include involvement of the orbit or the central nervous system, and vascular involvement. Despite surgical and antifungal treatment, mortality raises up to 75%. We report the case of a female patient with a diagnosis of diabetic ketoacidosis and signs of unilateral ophthalmoplegia, which led to the study with magnetic resonance imaging of the central nervous system, finding signs of sinusitis, meningitis, and cerebritis. Initial microbiological studies were negative, and biomarkers such as serum galactomannan and Cryptococcus antigen were also negative. After surgical management and the identification of Aspergillus flavus and Rhizopus spp. in sinus tissue, the patient received treatment with posaconazole and after two months of follow-up she presented clinical improvement. Dual fungal infection and infection by A. flavus are uncommon and clinically relevant entities, with no cases previously reported in our country, therefore this corresponds to a case of clinical interest.

Fungal decomposition of chicken-feather waste in submerged and solid-state fermentation / Decomposição fúngica de resíduos de pena de frango em fermentação submersa e de estado sólido

Abstract Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.

Resumo A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.

Fungal decomposition of chicken-feather waste in submerged and solid-state fermentation / Decomposição fúngica de resíduos de pena de frango em fermentação submersa e de estado sólido

Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.

A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.

Fungal decomposition of chicken-feather waste in submerged and solid-state fermentation

Abstract Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.

Resumo A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.

Bio-valorization of palm oil mill effluent waste for the potential production of renewable biomass fuel pellets

Aims@#The primary aim of this study was to utilize abundant palm oil mill effluent (POME) waste and turn it into a value-added product of biomass fuel with high calorific energy value (CEV) via fermentation and drying process, then simultaneously reduce abundant liquid waste.@*Methodology and results@#POME is available abundantly in Malaysia and only a small portion of it is utilized to produce other value-added products. In this study, fermentation of POME in the presence of bacteria (Lysinibacillus sp.) and fungus (Aspergillus flavus) separately at 37 °C, 180 rpm for 5 days, followed by overnight oven-drying at 85 °C was conducted. Four fermentation medium conditions were performed, viz.: (1) autoclaved POME, (2) autoclaved POME with the addition of Lysinibacillus sp., (3) autoclaved POME with the addition of A. flavus and (4) POME as it is (non-sterile).@*Conclusion, significance and impact of study@#Among all conditions, fermentation utilizing autoclaved POME in the presence of A. flavus evinced the highest CEV of 25.18 MJ/kg. The fermentation in the presence of Lysinibacillus sp. strain revealed high COD and BOD removal efficiency of 59.20% and 320.44 mg/L as well as the highest reduction of oils and grease among other groups with the value of 15.84%. Future research directions are proposed for the elucidation of co-fermentation in the presence of both Lysinibacillus sp. and A. flavus.

Buffaloes (Bubalus bubalis) mycotic eye infection: reported herd cases in Thi-Qar province, Iraq / Infecção ocular micótica em búfalos (Bubalus bubalis): relato de casos em rebanho na província de Thi-Qar, Iraque

Buffaloes are one of the important farm animals in the south of Iraq and play an essential economical role mainly acting as dairy, meat, and draft animals. This study intended to diagnose buffalo mycotic eye infections in Thi-Qar province/Iraq. Some 250 buffaloes in the herd of 3,700 animals suffered from eye infections from December 2017 to November 2018. Eye swabs were collected from each infected eye of the affected buffaloes of both sexes before treatment. The animals were in different age groups. All samples were transferred to the laboratory in transfer media, and cultured on Sabouraud dextrose (SDA) agar with and without 0.05 g/mL and 0.4 g/mL chloramphenicol and cycloheximide, respectively. Later, the agars were incubated at 25oC and 37oC. The total percentage of eye infection was (6.75%), constituting (49.2%) mycotic infections. The predominant clinical manifestations that appeared on the infected buffaloes were eye inflammation represented by congestion, lacrimation, the opacity of cornea and edema, and reduced productivity of the infected animals. Different fungal isolates were identified from the samples including Aspergillus fumigates, Aspergillus flavus, Aspergillus niger, Penicillium spp., Alternaria spp., Fusarium spp., Candida spp., Cladosporium spp., Rhodotorula spp., Mucor spp. and Rhizopus spp. Calves buffaloes below one-year-old were more prone to mycotic infection than one-year-old or more. Additionally, male buffaloes were more susceptible to infection than females. In conclusion, this study isolated various types of fungus from the inflamed eyes of buffaloes. Fungal eye infection and the potential risk factors for fungal keratitis in buffaloes were also investigated. The study also approved the rapid diagnosis of fungi by direct microscopic detection and culture. The author recommends future studies including large numbers of the buffalo herd in Iraq to determine the epidemiology of this condition in the country.(AU)

Os búfalos são um dos animais de fazenda mais importantes no sul do Iraque e desempenham um papel econômico essencial, atuando principalmente na produção de leite, carne e como animal de tração. Este estudo objetivou diagnosticar infecções oculares micóticas em búfalos na província de Thi-Qar, Iraque. 250 búfalos no rebanho de um total de 3700 animais apresentaram infecção ocular durante o período compreendido entre dezembro de 2017 e novembro de 2018. Os esfregaços oculares foram colhidos dos olhos infectados dos búfalos afetados de ambos os sexos antes do tratamento. Os animais estavam em diferentes faixas etárias. Todas as amostras foram transferidas para o laboratório por meio de transferência e cultivadas em Ágar Sabouraud e Dextrose (SDA) com e sem 0,05 g/mL e 0,4 g/mL de cloranfenicol e cicloheximida, respectivamente. Posteriormente, os ágares foram incubados a 25ºC e 37ºC. A porcentagem total de infecção ocular foi de 6,75%, representando 49,2% de infecção micótica. As manifestações clínicas predominantes nos búfalos infectados foram inflamação ocular com congestão, lacrimejamento, opacidade da córnea e edema. Os animais acometidos também apresentaram redução de produtividade. Diferentes isolados de fungos foram identificados a partir das amostras, incluindo Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Penicillium spp., Alternaria spp., Fusarium spp., Candida spp., Cladosporium spp., Rhodotorula spp., Mucour spp. e Rizopus spp. Búfalos com menos de um ano de idade foram mais propensos a infecções micóticas do que com um ano ou mais. Além disso, os búfalos machos foram mais suscetíveis a infecção do que as fêmeas. Em conclusão, este estudo registrou o isolamento de vários tipos de fungos em olhos inflamados de búfalos. Além disso, a infecção ocular por fungos e os fatores de risco potenciais para ceratite fúngica em búfalos também foram observados. O estudo também aprovou o diagnóstico rápido de fungos por detecção microscópica direta e cultura. O autor recomenda outro estudo futuro, incluindo um grande número de rebanhos de búfalos no Iraque para determinar a epidemiologia desta condição no país.(AU)

In vitro antibacterial activity of endophytic fungus Aspergillus flavus IBRL-C8 from Senna siamea against Gram-bacteria

Aims@#To evaluate the antibacterial efficacy of ethyl acetate extract of Aspergillus flavus IBRL-C8 against Gram-positive and Gram-negative bacteria.@*Methodology and results@#In this experiment, an endophytic fungus which identified as A. flavus IBRL-C8 was extracted using ethyl acetate and methanol, from Senna siamea, prior to in vitro antibacterial test on eight Gram-bacteria. The results were significantly more enunciated to the ethyl acetate extract since the Gram-bacteria signified 9.0 to 20.0 mm of inhibition zones on Muller Hinton Agar (MHA) during disc diffusion assay. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the extract were ranged from 125-1000 µg/mL and 125-2000 µg/mL, respectively. Time-kill assay depicted the ethyl acetate extract of A. flavus IBRL-C8 exceptionally retarded methicillin-resistant Staphylococcus aureus (MRSA) and also manifested extended antibacterial activity. The maximum reduction in cell numbers occurred at 2MIC concentration (250 µg/mL) during the interval time of 16 h. The malformations noticed from microscopic observations where the transformation of structural annihilation from regular spherical morphology to non-spherical shape with an irregular surface and also disruption around the cell membrane when the MRSA treated with ethyl acetate extract of A. flavus IBRL-C8. @*Conclusion, significance and impact of study@#This study proposed the ethyl acetate extract of A. flavus IBRL-C8 as a potential antibacterial agent against MRSA infection, which can be useful in pharmaceutical application.

Production and optimization of enzyme xylanase by Aspergillus flavus using agricultural waste residues

This study aims to isolate xylanase-producing fungi, i.e., Aspergillus flavus, isolated from the litter of theOrchha forest, and efforts are made to culture fungi on cheaper agricultural substrates, i.e., wheat bran and corncobs. After isolation, the xylanolytic activity was tested on the malt extract agar culture medium. Optimizationof growth parameters were also carried out with wheat bran and corn cobs. It was found that maximum xylanasewas produced on the sixthand eighth days of incubation, with pH of 6.0 and 7.0, and substrates amount for corncobs and wheat bran of 16 and 18 mg/ml, respectively. Temperature of 30°C, peptone concentration of 0.5 mg/ml, and yeast extract concentration of 0.75 mg/ml were the same for both substrates. The enzyme producedthrough optimized conditions was assayed for its maximum activity. It was found that the enzyme showedmaximum activity at 15 and 30 minutes of incubation, and at 60° and 55°C temperature for corn cobs andwheat bran, respectively. The pH and substrate (oat spelt xylan) concentrations were optimized at 5.5 and 15mg/ml, respectively. In. this study, the production of xylanase with the use of cheaper agricultural waste, suchas wheat bran and corn cobs, will not only reduce the cost of production but will also help in their eradicationfrom the environment.

Multiple invasive aspergillus brain abscess in an immunocompetent patient without any history of trauma

Aspergillus is a fungus found in the environment. In an immunocompetent person, inhalation of spores may cause localized infection. In immune compromised patients, these fungi can cause life-threatening invasive infections which has high morbidity and mortality. Invasive aspergillosis has a poor prognosis. Intracranial aspergillosis is an extremely rare manifestation of invasive aspergillosis in immunocompetent individuals. A case of 60-year-old immunocompetent male is reported who had multiple Aspergillus brain abscess.

Antimicrobial activities of endophytic fungi residing in Aloe vera against diabetic wound pathogens

Aims@#Endophytic fungi are organisms that colonize living plant tissues without causing significant symptoms of diseases. They are known as the repository of bioactive compounds. This study was aimed to isolate endophytic fungi associated with Aloe vera and to assess their antimicrobial activity against the diabetic wound pathogens that result to lower limb amputations. @*Methodology and results@#In this study, 84 endophytic fungi were isolated from A. vera by using 5% sodium hypochlorite and 70% ethanol as surface sterilants. On preliminary antimicrobial screening by agar plug assay, 12 (14.3%) isolates showed significant antagonistic activity on the test microorganisms. Two of the endophytes were identified as Aspergillus flavus and Colletotrichum gloeosporioides through morphological and molecular techniques. The fungal cultures fermented in shake flask were extracted subsequently with hexane, dichloromethane, ethyl acetate, and n-butanol. Ethyl acetate and dichloromethane extracts of A. flavus and C. gloeosporioides inhibited all test bacteria. The compounds in the extracts were seemed to be semi-polar in nature. On broth microdilution assay, minimal inhibitory and minimal lethality concentration of ethyl acetate extracts of the fungi were found in the range of 0.63−2.50 and 1.25−5.00 mg/mL, respectively. @*Conclusion, significance and impact of study@#This study reveals that A. vera harbours a wide diversity of endophytic fungi. A. flavus and C. gloeosporioides showing broad spectrum antimicrobial activity on wound pathogens may be potential sources of bioactive compounds. Further investigations should be conducted to isolate and identify the antimicrobial compound produced by these fungal isolates.

A comparative study on enzyme production ability of microorganism producing γ-aminobutyric acid in processing of Sojae Semen Praeparatum at different pH and temperatures / 中草药

Objective: To lay a foundation for elucidating the mechanism of microbial regulation of γ-amino butyric acid (GABA) formation in the processing of Sojae Semen Praeparatum. Methods: The ability of glutamic acid decarboxylase and protease (neutral protease, alkaline protease and acid protease) production of Bacillus subtilis, Enterococcus avium, Enterococcus faecium, Bacillus amyloliquefaciens, Aspergillus niger, Aspergillus flavus, Aspergillus tamarii, Penicillium citrinum, Cladosporium tenuissimum, Phanerochaete sordida, Rhizopus oryzae, Sporidiobolus salmonicolor in different pH and temperature conditions were determined by Berthelot colorimetry and Folin phenol method. Results: The GAD and protease activities of the twelve strains were stronger at pH 5-7 and temperature 28-37 ℃. The highest GAD enzyme activity from A. flavus was 41.97 U/h, the optimum pH was 7 and the temperature was 28 ℃, followed by S. salmonicolor, A. niger, R. oryzae, and B. subtilis, respectively. The enzyme activities were 29.04, 25.78, 22.42 and 19.43 U/h. The highest neutral protease activity of B. subtilis was 24.80 U/mL, the optimum pH was 7 and the temperature was 37 ℃, followed by B. amyloliquefaciens, R. oryzae and E. avium. The enzyme activities were 16.86, 12.51 and 9.18 U/mL. The highest alkaline protease activity of B. amyloliquefaciens was 13.29 U/mL, the optimum pH was 7 and the temperature was 34 ℃, followed by B. subtilis and R. oryzae. The enzyme activities were 8.86 and 6.20 U/mL, respectively. The ability of 12 kinds of microorganisms to produce acid protease was generally poor. Conclusion: The optimal pH and temperature of the 12 kinds of microorganisms selected for this experiment are basically consistent with the natural processing of Sojae Semen Praeparatum. Among them, fungi have stronger GAD production capacity and bacteria have stronger neutral protease production capacity. The high GABA concentration of Sojae Semen Praeparatum is caused by the joint action of multiple strains.

Research progress on growth of Aspergillus flavus and its toxins production controlled by plant essential oils and its active ingredients / 中草药

Aspergillus flavus is a pathogenic fungus, which can easily contaminate Chinese herbal medicines, food, and agricultural commodities. Aflatoxin (AFT) produced by the secondary metabolism of A. flavus caused serious threats to the health of people and animals due to its carcinogenic and highly toxic characteristics. Therefore, the search for inhibitors to A. flavus and its toxins has been widely concerned in the world. Essential oil is a natural antifungal agent extracted and highly concentrated from higher plants. It has the advantages of good antifungal effect, low pollution, and relatively safe to human body. Some kinds of them, including Thymus vulgaris, Origanum virens, Perilla frutescens, Foeniculum vulgare, and so on, displayed their antifungal activities. Specifically, active ingredients such as alcohols, aldehydes, and phenols have relatively good antifungal effects. The research progress on pathogenicity of A. flavus and AFT and the prevention and control by essential oils are reviewed in this article, which provided a reference for further exploring the antifungal mechanism of essential oils and developing natural and green antifungal products.

Study on Inhibitory Effect of Volatile Oils in 16 Plants on Aspergillus flavus of Platycladi Semen / 中国实验方剂学杂志

Objective::To select volatile oils from 16 species of plants (Cymbopogon citratus, Pelargonium graveolens, Pinus tabulieformis, Litsea cubeba, Mentha haplocalyx, Zingiber officinale, Syzygium aromaticum, Curcuma longa, Zanthoxylum bungeanum, Cinnamomum cassia, Ocimum basilicum, Rosmarinus officinalis, Zanthoxylum schinifolium, Zanthoxylum armatum, Illicium verum, Myristica fragrans) that have good inhibitory effect on the growth of Aspergillus flavus. Method::Aspergillus flavus was isolated from the surface of Platycladi Semen medicinal materials by plate culture method. The volatile oils of 16 plants were extracted by steam distillation. The colony diameter of Aspergillus flavus was determined by fumigation of filter paper, and the effect of volatile oils on the growth of Aspergillus flavus was systematically studied. Result::Aspergillus flavus was successfully isolated from Platycladi Semen by means of morphological, microscopic and DNA barcoding identification methods, the bacteriostatic rates of the above 16 kinds of volatile oils against Aspergillus flavus were 2.93%, 0.05%, 0.37%, 76.07%, 0.34%, 0.15%, 50.05%, 8.51%, 1.43%, 58.20%, 0.07%, 2.60%, 8.73%, 100.00%, 52.62%, 0.07%, respectively. Conclusion::The volatile oils of 16 plants all have different degrees of antibacterial activities for Aspergillus flavus, and volatile oils of Zanthoxylum armatum, Litsea cubeba and Cinnamomum cassia have good inhibitory effect. This study can provide a theoretical basis for the prevention and control of Aspergillus flavus in the growth and storage of Platycladi Semen, and provide a basis for further research on plant volatile oil as bacteriostatic agents in the storage process of traditional Chinese medicine.

El factor de transcripción bZIP Afap1 afecta al estrés oxidativo y la biosíntesis de aflatoxinas en Aspergillus flavus / The bZIP transcription factor Afap1 mediates the oxidative stress response and aflatoxin biosynthesis in Aspergillus flavus

Abstract Aflatoxin is a carcinogenic secondary metabolite produced mainly by Aspergillus flavus and Aspergillus parasiticus, which can seriously endanger the health of humans and animals. Oxidative stress is a common defense response, and it is known that reactive oxygen species (ROS) can induce the synthesis of a series of secondary metabolites, including aflatoxin. By using mutants lacking the afap 1 gene, the role of afap 1 gene in oxidative stress and aflatoxin synthesis was assessed. The growth of the mutant strains was significantly inhibited by the increase in the concentration of H2O2, inhibition was complete at 40mmol/l. However, in the quantitative analysis by HPLC, the concentration of AFB1 increased with the increased H 2O 2 until 10mmol/l. Following an analysis based on the information provided by the NCBI BLAST analysis, it was assumed that Afap1, a basic leucine zipper (bZIP) transcription factor, was associated with the oxidative stress in this fungus. Treatment with 5mmol/l H 2O 2 completely inhibited the growth of the mutant strains in afap 1 but did not affect the growth of the CA14PTs strain (non-mutant strain). In addition, the concentration of AFB 1 in the mutant strains was approximately V of that observed in the CA14PTs strain. These results suggested that Afap1 plays a key role in the regulation of oxidative stress and aflatoxin production in A. flavus. ©2018 Published by Elsevier España, S.L.U. on behalf of Asociación Argentina de Microbiología. This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/ licenses/by-nc-nd/4.0/).

Resumen La aflatoxina es un metabolito secundario cancerígeno producido principalmente por Aspergillus flavus y Aspergillus parasiticus, que pone en riesgo grave a la salud de los humanos y los animales. El estrés oxidativo es una respuesta de defensa común, y es sabido que las especies reactivas de oxígeno (ROS) pueden inducir la síntesis de una serie de metabolitos secundarios, incluida la aflatoxina. Empleando mutantes carentes del gen afap1 se evaluó el papel de Afap1 en el estrés oxidativo y la síntesis de aflatoxinas. El crecimiento de las cepas mutadas se vio significativamente inhibido con el aumento de la concentración de H 2O 2, la inhibición fue completa a 40mmol/l. Sin embargo, en el análisis cuantitativo por HPLC, la concentración de la aflatoxina AFBi aumentó con el aumento de la concentración de H 2O 2 hasta 10mmol/l. Tras un análisis apoyado en la información provista por la herramienta NCBI BLAST, se supuso que Afap1, un factor de transcripción de la cremallera de leucina básica (bZIP), estaba asociado con el estrés oxidativo en este hongo. El tratamiento con 5mmol/l de H 2O 2 inhibió completamente el crecimiento de las cepas mutantes en afap1, pero no afectó el crecimiento de la cepa CA14PTs (cepa no mutada). Además, la concentración de AFB 1 en las cepas mutadas fue de aproximadamente 1/4 de la observada en CA14PTs. Estos resultados sugieren que Afap1 juega un papel clave en la regulación del estrés oxidativo y la producción de aflatoxinas en A. flavus.

Morphological and toxigenic study of Aspergillus flavus from indoor air

Aim: In the current investigation thirty four A. flavus cultures were isolated from indoor air samples of some farmers' house in coastal region of Odisha. Production of aflatoxin B1 and the pattern of formation of sclerotia by the isolates were studied in vitro. Methodology: Morphological study and production of sclerotia were carried out by observing fungal colonies on culture media. Conidiophores were identified under Phase contrast microscope. Detection of aflatoxin B1 was done by Thin Layer Chromatography technique. Results: The isolates exhibited variation in their colony characteristics and pattern of sclerotia formation. Overall 44% A. flavus isolates produced stable aflatoxin B1 and 41% were producers of sclerotia. About 32% A. flavus isolates produced both sclerotia and aflatoxin B1. Most of the non toxigenic isolates failed to form thick mycelia mat on β-CD-PDA media. Aflatoxin and sclerotia producing isolates were found more in coastal districts than inter-coastal regions. No significant variation in the pattern of sclerotia production was observed among isolates in less aerobic (sealed plates) and aerobic (unsealed plates) condition, however isolates had slightly more preference towards less aerobic environment for sclerotia production. The size of sclerotia was found to be inversely related to number of sclerotia. Interpretation: The presence of diverse A. flavus strains in farmers' indoor air, having the ability to produce either aflatoxin or sclerotia or both, may lead to serious health implications. This issue need to be addressed and a suitable way for proper management of indoor environment of farmers' need to be designed.

Transcriptome and proteome analyses of resistant preharvest peanut seed coat in response to Aspergillus flavus infection

BACKGROUND: The infection of peanut (Arachis hypogaea L.) seed coat by the pathogenic fungus Aspergillus flavus has highly negative economic and health impacts. However, the molecular mechanism underlying such defense response remains poorly understood. This study aims to address this issue by profiling the transcriptomic and proteomic changes that occur during the infection of the resistant peanut cultivar J11 by A. flavus. RESULTS: Transcriptomic study led to the detection of 13,539 genes, among which 663 exhibited differential expression. Further functional analysis found the differentially expressed genes to encode a wide range of pathogenesis- and/or defense-related proteins such as transcription factors, pathogenesis-related proteins, and chitinases. Changes in the expression patterns of these genes might contribute to peanut resistance to A. flavus. On the other hand, the proteomic profiling showed that 314 of the 1382 detected protein candidates were aberrantly expressed as a result of A. flavus invasion. However, the correlation between the transcriptomic and proteomic data was poor. We further demonstrated by in vitro fungistasis tests that hevamine-A, which was enriched at both transcript and protein levels, could directly inhibit the growth of A. flavus. Conclusions: The results demonstrate the power of complementary transcriptomic and proteomic analyses in the study of pathogen defense and resistance in plants and the chitinase could play an important role in the defense response of peanut to A. flavus. The current study also constitutes the first step toward building an integrated omics data platform for the development of Aspergillus-resistant peanut cultivars

Desarrollo y capacidad de remoción de los pesticidas atrazina, clorpirifós y endosulfán por parte de cepas de Aspergillus sección Flavi no toxigénicas / Development and removal ability of non-toxigenic Aspergillus section Flavi in presence of atrazine, chlorpyrifos and endosulfan

This study evaluated the in vitro effect of three concentrations of atrazine, chlorpyrifos and endosulfan on the growth parameters of four non-toxigenic Aspergillus section Flavi strains. The ability of the strains to remove these pesticides in a synthetic medium was also determined. Growth parameters were measured on soil extract solid medium supplied with 5,10 and 20mg/l of each pesticide, and conditioned to -0.70, -2.78, -7.06 and -10.0 water potential (MPa). Removal assays were performed in Czapek Doc medium (CZD) supplied with 20mg/l of each pesticide under optimal environmental conditions (-2.78 of MPa and 25 °C). The residual levels of each pesticide were detected by the reversed-phase HPLC/fluorescence detection system. The lag phases of the strains significantly decreased in the presence of the pesticides with respect to the control media. This result indicates a fast adaptation to the conditions assayed. Similarly, the mycelial growth rates in the different treatments increased depending on pesticide concentrations. Aspergillus oryzae AM 1 and AM 2 strains showed high percentages of atrazine degradation (above 90%), followed by endosulfan (56 and 76%) and chlorpyrifos (50 and 73%) after 30 days of incubation. A significant (p <0.001) correlation (r = 0.974) between removal percentages and growth rate was found. This study shows that non-toxigenic Aspergillus section Flavi strains from agricultural soils are able to effectively grow in the presence of high concentrations of atrazine, chlorpyrifos and endosulfan under a wide range of MPa conditions. Moreover, these strains have the ability to remove high levels of these pesticides in vitro in a short time.

En este estudio se evaluó los efectos in vitro de 3 concentraciones de atrazina, clorpirifós y endosulfán sobre los parámetros de crecimiento de 4 cepas no toxigénicas de Aspergillus sección Flavi. También se evaluó la capacidad de las cepas de remover los pesticidas. Los parámetros de crecimiento se ensayaron en medio agar extracto de suelo suplementado con 5, 10 y 20mg/l de cada pesticida y acondicionado a -0.70, -2.78, -7.06 y -10.0 de potencial de agua (MPa). Los ensayos de remoción se realizaron en medio Czapek Dox con 20mg/l de cada pesticida bajo condiciones óptimas de crecimiento (-2.78 de MPa y 25 °C). Los niveles residuales de atrazina, clorpirifós y endosulfán se detectaron en un sistema HPLC con detección por fluorescencia. La fase de latencia de las cepas disminuyó significantemente en presencia de los pesticidas, indicando una rápida adaptación a dichas condiciones. La velocidad de crecimiento se incrementó considerablemente dependiendo de la concentración de pesticida. Las cepas Aspergillus oryzae AM1 y AM2 mostraron porcentajes elevados de degradación de atrazina (aproximadamente el 90%), seguidos por endosulfán (56 y 76%) y clorpirifós (50 y 73%). Se observó una correlación (r = 0.974) significante (p <0.001) entre el porcentaje de pesticida removido y la velocidad de crecimiento. Este estudio muestra que cepas no-toxigénicas de Aspergillus sección Flavi aisladas de suelos agrícolas desarrollan eficientemente en presencia de altas concentraciones de atrazina, clorpirifós y endosulfán en un amplio rango de MPa. Además, presentan capacidad de remover in vitro altos niveles de pesticidas en corto tiempo.

Research Progress on Prevention and Control Measures of Aflatoxin in Chinese Medicinal Materials / 中国实验方剂学杂志

At present,traditional Chinese medicine(TCM) has attracted more and more attention from the international community.The demand for TCM is increasing in the world.The hidden dangers of potential quality and safety of TCM are also becoming increasingly prominent.Aflatoxin contamination has become one of the important factors affecting the safety of Chinese herbal medicines,and it will fundamentally affect human health and life safety.A variety of methods are used to reduce aflatoxins,however,there are few suitable methods that can be widely used in the cost-effective and large-scale promotion of Chinese herbal medicines.Therefore,it is of great significance to continue to study measures to solve the pollution problems of Aspergillus flavus and its toxins.This article summarizes the hazards and contamination status of aflatoxin,the prevention and control of the growth of A. flavus, and the measures for reducing aflatoxin,and looks ahead to the future prevention and control of A. flavus and its toxins,aiming at providing ideas for the pollution problem of A. flavus and its toxin,to ensure the quality of Chinese herbal medicines,so as to ensure clinical safety medication.

Investigation of aflatoxins,mycobiota,and toxigenic fungi during post-harvest handling of Ziziphi Spinosae Semen / 中国中药杂志

Ziziphi Spinosae Semen is one of the Chinese herbal medicine being susceptible to aflatoxins contamination. To investigate the sources of aflatoxins contamination and toxigenic fungi species on Ziziphi Spinosae Semen,32 samples were collected from multiple steps during the post-harvest processing in this study. Aflatoxins in these samples were determined by immunoaffinity column and HPLC coupled with post-column photochemical derivatization. The dilution-plate method was applied to the fungi isolation. The isolated fungi strains were identified by morphological characterization and molecular approaches. The results showed that aflatoxins were detected in 28 samples from every step during the processing of Ziziphi Spinosae Semen. Three samples were detected with aflatoxin B_1 and 2 samples with both aflatoxin B_1 and total aflatoxin exceeding the limit of Chinese Pharmacopoeia. Especially the samples from the washing step,with the highest detected amounts of AFB_1 and AFs were reached 94. 79,121. 43 μg·kg~(-1),respectively. All 32 samples were contaminated by fungi. The fungal counts on the newly harvested samples were 2. 20 × 10~2 CFU·g~(-1). Moreover,it increased as tphreocessing progresses,and achieved 1. 16×10~6 CFU·g~(-1) after washing. A total of 321 isolates were identified to 17 genera. Aspergillus flavus was the main source of aflatoxins during the processing and storage of Ziziphi Spinosae Semen. One isolate of A. flavus was confirmed producing AFB_1 and AFB_2. The fungal count was significantly increased by composting,and Aspergillus was the predominant genus after shell breaking. The contamination level of aflatoxins was increased by composting and washing.

Study on the surface fungus diversity of commercially Platycladi semen based on high-throughput sequencing technology / 药学学报

The structural composition of the surface fungal community of commercially Platycladi semen was analyzed to reveal the surface fungal biodiversity and structural differences. Platycladi semen was collected from Henan, Shandong and Hong Kong, their DNA was extracted, ITS fragments in DNA were amplified by PCR. Miseq was sequenced on Illumina Hiseq 2500 platform after the PCR products were qualified for quality inspection. The sequence OTU cluster was obtained and bioinformatics analysis was carried out. Microbial communities were not observed in the eyes of the Platycladi semen in the three regions. Sequencing results showed that the surface microbial community had high biodiversity, but there were significant differences in species composition. Seven samples o Platycladi semen obtained 345 947 valid sequences, which were divided into 267 OTUs, 3 phylums. 18 classes, 40 orders, 82 families, 120 genus, 191 species fungi. At the genus level, Aspergillus is the dominant species, accounting for the highest proportion, reaching (93.36 ± 6.01)%. Seven samples were contaminated by Aspergillus flavus, and the pollution levels were 14.58%, 15.98%, 17.64%, 16.44%, 0.97%, 23.39% and 18.86%. Except sample No. 5, Aspergillus cibarius was the most abundant, the other six samples were Aspergillus niger, Aspergillus fumigatus and Aspergillus flavus as the core microflora. By analyzing the diversity of fungi distribution in different habitats, we can fully understand the fungi on the surface of Platycladi semen, lay a foundation for early risk warning of Aspergillus flavus contamination and its aflatoxin contamination, and provide a theoretical basis for the quality and safety of Platycladi semen.