Improvement of the production and quality of Cordyceps javanica conidia for the control of Diaphorina citri adults.

This study aimed to evaluate the use of palm kernel meal (PKM) in the traditional solid-state fermentation system to improve the production and quality of Cordyceps javanica conidia. The impact of PKM was determined by measuring conidia yield, viability, hydrophobicity, shelf life, and conidia pathogenicity against Diaphorina citri adults. By supplementing rice grains with 5% palm kernel meal increased the conidial yield by up to 40%, without compromising conidia viability and hydrophobicity. In addition, conidia caused higher levels of mortality by mycosis against D. citri adults (90%), relative to conidia harvested from rice (52%). The conidia recovered from rice/palm kernel meal mixtures also retained viability greater than 90% after storage for 10 months at 4 °C, while the conidia produced on rice reached 80%. Thus, conidia produced in the presence of palm kernel meal can be consumed immediately or in the medium term. Some process advantages of the palm kernel meal as co-substrate in the traditional production system of C. javanica are also mentioned. These results are attractive for improving the mycoinsecticide production process, with excellent cost-benefit and minimal changes in infrastructure and process.

Optimization of cultural and nutritional conditions to enhance mycelial biomass of Cordyceps militaris using statistical approach.

Cordyceps militaris is a fungus with numerous therapeutic properties that has gained worldwide popularity due to its potential health benefits. The fruiting body of this mushroom is highly expensive and takes a longer time to produce, making mycelial a sustainable and cost-effective alternative. The study investigates and optimizes cultural and nutritional conditions to maximize mycelial biomass. The initial optimization was done by the conventional single-factor approach, followed by Plackett-Burman design to screen the most significant variables, with yeast extract, temperature, and glucose being the most significant, contributing 11.58%, 49.74%, and 27.98%, respectively, in mycelial biomass production. These variables were then optimized using response surface methodology (RSM) based on central composite design (CCD). The study observed that temperature and glucose had the highest impact on mycelial biomass, with p-values of 0.0128 and 0.0191, respectively. Under the optimized conditions, temperature 20 °C, glucose 2.5% (w/v), and yeast extract 0.8% (w/v), the maximal yield of mycelial biomass reached 547 ± 2.09 mg/100 mL, which was 1.95-fold higher than the yield in the basal medium. These findings suggest that optimizing the cultural and nutritional conditions can enhance mycelial biomass production of Cordyceps militaris, offering a sustainable and cost-effective source of this valuable fungus.

Isolation and molecular characterization of Cordyceps sp. from Bemisia tabaci (Hemiptera: Aleyrodidae) and pathogenic to Glycaspis brimblecombei (Hemiptera: Aphalaridae) / Isolamento e caracterização molecular de Cordyceps sp. de Bemisia tabaci e patogênico para Glycaspis brimblecombei (Hemiptera: Aphalaridae)

The Brazilian forestry sector stands out for its technology, forestry management practices, social and environmental responsibility and, mainly, for its high productivity and exotic pests can reduce it. The red gum lerp psyllid Glycaspis brimblecombei (Moore, 1964) (Hemiptera: Aphalaridae) is an important pest in Eucalyptus plantations. The parasitoid Psyllaephagus bliteus (Riek, 1962) (Hymenoptera: Encyrtidae), predatory bugs and entomopathogenic fungi such as Beauveria bassiana and Metarhizium anisopliae are the natural enemies and used in the biological control of the red gum lerp psyllid. The use of entomopathogenic fungi against exotic pests is increasing in the forestry sector and the prospecting and identification of fungus isolates is important for integrated pest management. The objective of this work was the isolation and molecular identification of Cordyceps spp. And to evaluate the pathogenicity of isolates, obtained from Bemisia tabaci (Gennadius, 1889) (Hemiptera: Aleyrodidae) adults, against to the red gum lerp psyllid G. brimblecombei. The fungi were isolated from B. tabaci adults found in soybean and tomato crops and molecularly identified. The conidia obtained were suspended in solution with Tween 80 (0.1%) at a concentration of 1.0 × 108 conidia/mL and sprayed on ten G. brimblecombei nymphs per Eucalyptus leaf cut and placed on a hydroretentive gel inside per Petri dishes as a replication. The number of dead insects was quantified, daily, for seven days, and transferred to humid chambers. Cordyceps javanica (LCBPF 11) and C. fumosorosea (LCBPF 12 and LCBPF 63) were identified with a molecular analysis and all isolates were pathogenic to the insects and indicates that they could be used to manage G. brimblecombei and adds to reports that, normally, fungi cause greater mortality on insects of the same order as that from which they were isolated.

O setor florestal brasileiro se destaca pela tecnologia, práticas de manejo florestal, responsabilidade social e ambiental e, principalmente, alta produtividade, mas pragas exóticas podem reduzir isto. O parasitoide Psyllaephagus bliteus (Riek, 1962) (Hymenoptera: Encyrtidae), percevejos predadores e fungos entomopatogênicos como Beauveria bassiana e Metarhizium anisopliae são inimigos naturais usados no controle biológico do psilídeo-de-concha do eucalipto Glycaspis brimblecombei (Moore, 1964) (Hemiptera: Aphalaridae), praga em plantios de eucalipto. A utilização de fungos entomopatogênicos contra pragas exóticas vem aumentando no setor florestal e a prospecção e identificação de isolados fúngicos é importante para o manejo integrado de pragas. O objetivo deste estudo foi o isolamento e identificação molecular de Cordyceps spp. e avaliar a patogenicidade dos isolados de Cordyceps javanica (LCBPF 11) e Cordyceps fumosorosea (LCBPF 12 e LCBPF 63), obtidos de adultos de Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae), ao psilídeo-de-concha G. brimblecombei. Os fungos foram isolados de adultos de B. tabaci encontrados em culturas de soja e tomate e identificados molecularmente. Os conídios foram suspensos em solução com Tween 80 (0,1%) na concentração de 1,0 x 108 conídios/mL e pulverizados sobre dez ninfas de G. brimblecombei por folha de Eucalyptus cortada e colocada sobre gel hidrorretentor no interior de placas de Petri por repetição. O número de insetos mortos foi quantificado, diariamente, por sete dias e transferidos para câmaras úmidas. Cordyceps javanica (LCBPF 11) e C. fumosorosea (LCBPF 12 e LCBPF 63) foram identificados por análise molecular e todos os isolados foram patogênicos para os insetos. Isto indica serem importantes para o manejo de G. brimblecombei e confirma relatos de que, normalmente, fungos causam maiores mortalidades em insetos da mesma ordem daquela da qual foram isolados.

Enhanced production of cordycepin under solid-state fermentation of Cordyceps militaris by using combinations of grains/substrates.

This manuscript deals with cordycepin, an interesting secondary compound produced from entomopathogenic fungus, Cordyceps. It has attracted commercial interest due to its immense pharmacological importance beneficial to human health. In this study, the contents of cordycepin and its derivatives, like adenine and adenosine, were evaluated through solid-state fermentation using combinations of various grains as substrate. Treatment with grain combination numbers 2, 7, 8, and 9 exhibited higher cordycepin content (1.621, 1.929, 1.895, and 1.996 mg/g cordycepin, respectively) than control (rice). The grain combination number 7 exhibited significantly higher adenine content (700 mg/g) than the control and all other combinations. Treatments with grain combination numbers 2, 5, and 7 exhibited higher adenosine content (2.719, 2.938, and 3.392 mg/g, respectively); however, no significant increase in adenosine content was noted in any treatments. The biomass including fresh mycelium and fruit body was found higher in grain combination numbers 7 and 9, leading to enhanced cordycepin content. Overall, the increase in the fresh biomass significantly enhanced cordycepin accumulation. The level of cordycepin was recorded as higher than that of its derivatives, adenosine and adenine. The grain combination of rice, wheat, jowar, bajra, and sugarcane bagasse added to basal medium exhibited the highest cordycepin content and was found suitable for solid-state fermentation of Cordyceps militaris. To our understanding, the present study is the first to use combinations of cereals for the production of cordycepin from C. militaris.

The effect of low-dose Cordyceps on ischemia-reperfusion injury of the kidney in rats.

OBJECTIVE: None of studies have been conducted in terms of demonstrating the same effect with the low dose in cordycepin. In our study, we analyzed the histopathological and biochemical changes of low-dose Cordycepin(c) on a rat model in the kidney. MATERIALS AND METHODS: Twenty-four male Wistar Albino rats were randomly allocated to three groups (n = 8): the sham-control group (Group 1), the renal I/R-untreated (Group 2) group, and the I/R-C-treated (Group 3) group. Cordyceps was administered intraperitoneally at 5 mg/kg twice. Renal histological changes were compared and the relevant parameters of oxidative stress and inflammation were detected. RESULTS: In blood and tissue biochemistry, it was observed that IL-1 Beta, IL 6, TNF alpha, MDA, TOS, and OSI increased in Group 2 and decreased in Group 3. It was determined that TAS values were increased in Group 3, and decreased in Group 2. In the histopathological evaluation, while Group 1 was evaluated as normal, significant kidney damage was detected in Group 2. It was determined that there was a significant decrease in kidney damage in Group 3. CONCLUSION: These results suggest that low dose Cordycepin was as effective as normal dose on renal ischemic reperfusion and reduction of damage.

OBJETIVO: Ninguno de los estudios se ha realizado en términos de demostrar el mismo efecto con la dosis baja de cordicepina. En nuestro estudio, analizamos los cambios histopatológicos y bioquímicos de Cordycepin(c) en dosis bajas en un modelo de rata con isquemia-reperfusión (I/R) inducida en el riñón. MATERIALES Y MÉTODOS: Veinticuatro ratas macho Wistar Albino se asignaron al azar a tres grupos (n = 8): el grupo de control simulado (Grupo 1), el grupo sin tratamiento I/R renal (Grupo 2) y el grupo tratado con I/R-C (Grupo 3). Cordyceps se administró por vía intraperitoneal a 5 mg/kg dos veces. Se compararon los cambios histológicos renales y se detectaron los parámetros relevantes de estrés oxidativo e inflamación. RESULTADOS: En bioquímica sanguínea y tisular se observó que IL-1 Beta, IL 6, TNF alfa, MDA, TOS y OSI aumentaron en el Grupo 2 y disminuyeron en el Grupo 3. Se determinó que los valores de TAS aumentaron en el Grupo 3, y disminuyó en el Grupo 2. En la evaluación histopatológica, mientras que el Grupo 1 fue evaluado como normal, se detectó daño renal significativo en el Grupo 2. Se determinó que hubo una disminución significativa del daño renal en el grupo 3. CONCLUSIÓN: Estos resultados sugieren que la cordicepina en dosis bajas fue tan efectiva como la dosis normal en la reperfusión isquémica renal y la reducción del daño.

Entomopathogenic Fungi, Isolated From Soils and Bemisia tabaci (Hemiptera: Aleyrodidae) Adults, to Manage the Eucalyptus Red Gum Lerp Psyllid Glycaspis brimblecombei (Hemiptera: Aphalaridae).

The parasitoid Psyllaephagus bliteus Riek (Hymenoptera: Encyrtidae), entomopathogenic fungi, and chemical insecticides are the main strategies to manage the eucalypts pest Glycaspis brimblecombei Moore. The objective of this study was to isolate and to identify entomopathogenic fungi, collected from Bemisia tabaci Gennadius adults in soybean and tomato crops, and from soil samples in eucalypts, soybean, and native forest areas to evaluate their potential to manage G. brimblecombei. Twelve Beauveria and Cordyceps isolates were selected and compared with the commercial products Boveril Beauveria bassiana Bals. (Hypocreales: Cordycipitaceae), Metarril Metarhizium anisopliae Metschn. (Hypocreales: Clavicipitaceae), and Octane Cordyceps fumosorosea Wize (Hypocreales: Cordycipitaceae) and their respective strains. The fungal isolates were diluted in 0.1% aqueous Tween 80 at a concentration of 1.0 × 108 conidia/ml and sprayed on the G. brimblecombei nymphs with or without lerps. Pest mortality was higher and the TL50 and TL90 lower with the isolates LCBPF 11 C. javanica Frieder. & Bally (Hypocreales: Cordycipitaceae), LCBPF 12 (C. fumosorosea), and LCBPF 67 (C. fumosorosea) from B. tabaci adults. Fungi of the genera Beauveria and Cordyceps developed and caused high mortality of G. brimblecombei nymphs with lerps. The B. bassiana, C. cateniannulata Liang (Hypocreales: Cordycipitaceae), C. fumosorosea, C. javanica, and M. anisopliae isolates showed potential to manage G. brimblecombei. The lerp of this insect enhances entomopathogenic fungus development as a source of inoculum accelerating G. brimblecombei nymph mortality. Entomopathogenic fungi isolated from insects and soils are effective against G. brimblecombei and the presence of the lerp of this insect increases the effectiveness of its control.

Cordyceps militaris Inhibited Angiotensin-Converting Enzyme through Molecular Interaction between Cordycepin and ACE C-Domain.

One of the most important therapeutic modalities for the management of hypertension is the inhibition of the angiotensin-converting enzyme (ACE). Cordyceps militaris has received substantial attention because to its therapeutic potential and biological value. To gather information about the antihypertensive properties of C. militaris, the ACE inhibitory activity was evaluated. An ethanolic extract of the fruiting body of C. militaris was obtained, and the extract was separated by UHPLC method with a fluorescence detector for the quantification of cordycepin and adenosine. The ethanolic extract had a considerably higher cordycepin level. Additionally, an in vitro kinetic analysis was carried out to find out how much C. militaris extract inhibited ACE. This extract exhibited non-competitive inhibition on ACE. The Ki value of the C. militaris extract against ACE was found to be 8.7 µg/mL. To the best of our knowledge, this is the first report of the analysis of a protein cavity together with molecular docking carried out to comprehend the intermolecular interactions between cordycepin and the ACE C-domain, which impact the spatial conformation of the enzyme and reduce its capacity to break down the substrate. According to a molecular docking, hydrogen bonding interactions between the chemicals and the ACE S2' subsite are primarily responsible for cordycepin inhibition at the ACE C domain. All these findings suggest that C. militaris extract are a kind of natural ACE inhibitor, and cordycepin has the potential as an ACE inhibitor.

Enhanced production of cordycepic acid from Cordyceps cicadae isolated from a wild environment.

Cordyceps acid is an active component of Cordyceps cicadae and has a variety of medicinal uses, including anti-tumor effects, the prevention of cerebral hemorrhaging and myocardial infarction, and the inhibition of a wide range of bacteria. The objectives of this study were to identify C. cicadae fungi and optimize the culture conditions to obtain a high yield of cordycepic acid. First, a wild C. cicadae was identified by morphological observation and rDNA sequence analysis. Secondly, the optimal fermentation conditions were determined using a single-factor method, a Plackett-Burman design, and a Box-Behnken response surface. Finally, using the yield of fruit bodies and the content of cordyceps acid as indices, combined with a single-factor experiment and a response surface design, the best combination of conditions for cultivation was determined. The results showed that the best combination was as follows: sucrose 2%, tryptone 2%, KH2PO4 0.4%, MgSO4·7H2O 0.4%, an initial pH of the fermentation liquid of 7.0, 5% inoculum, fermentation for 4.5 d, a ratio of medium to liquid of 1:1.7, illumination intensity 150 Lux, illumination time 15 h per day, and 70% humidity. The content of cordycepic acid in the fruiting bodies developed in cultivation was 2.07-fold higher than that in the wild C. cicadae. This study provides a theoretical basis for the large-scale cultivation of C. cicadae with a high concentration of cordycepic acid.

Diversity of entomopathogenic fungi from soils of eucalyptus and soybean crops and natural forest areas / Diversidade de fungos entomopatogênicos em solos de culturas de eucalipto e soja e área de mata nativa

Soils present high fungal diversity, including entomopathogenic species. These fungi are used in pest control, providing easy production, multiplication, application, and dispersion in the field. The objective of the present study was to evaluate entomopathogenic fungal diversity in soils from eucalyptus and soybean crops and natural forest areas. These fungi were isolated using the "Bait Method" with Tenebrio molitor (Linnaeus, 1758) (Coleoptera: Tenebrionidae) larvae from 10 soil samples per area, collected at 10 cm deep in a zig-zag pattern. The isolated entomopathogenic fungi were cultivated in Petri dishes using PDA medium and their mycelia separated after seven days of incubation in a BOD-type chamber. Species of Aspergillus, Beauveria, Cordyceps, Fusarium, Metarhizium, Penicillium and Purpureocillium were identified. The "Bait Method" with T. molitor larvae is efficient to isolate entomopathogenic fungi with higher diversity from soils of the natural forest than the cultivated area.

A diversidade de fungos, incluindo espécies entomopatogênicas, é alta nos solos. Esses fungos são utilizados no manejo de pragas com facilidade de produção, multiplicação, aplicação e dispersão no campo. O objetivo foi avaliar a diversidade de fungos entomopatogênicos em solos de culturas de eucalipto e soja e áreas de mata nativa. Fungos entomopatogênicos foram isolados pelo "Bait Method" com larvas de Tenebrio molitor (Linnaeus, 1758) (Coleoptera: Tenebrionidae) de 10 amostras de solo por área, coletadas a 10 cm de profundidade em zig-zag. Os fungos isolados foram cultivados em três placas de Petri em meio BDA e seus micélios separados após sete dias de incubação em câmara tipo BOD. Fungos dos gêneros Aspergillus, Beauveria, Cordyceps, Fusarium, Metarhizium, Penicillium e Purpureocillium foram identificados. O "Bait Method" com larvas de T. molitor é eficiente para isolar fungos entomopatogênicos com maior diversidade em solos de área de mata nativa que naqueles com culturas de eucalipto e soja.

Microbial Control of Leptopharsa heveae Drake & Poor (Hemiptera: Tingidae) in Brazilian Rubber Tree Plantations: a Brief Historical Account and Identification of Entomopathogenic Fungi by Means of Multigene Phylogeny.

Over the last decades, a few fungal species have been mentioned in the literature associated with the rubber tree lace bug, Leptopharsa heveae Drake & Poor (Hemiptera: Tingidae). The rubber plantation area treated with these biocontrol agents has been steady since the 1990s, estimated in 10,000-15,000 ha per year. A few large-scale rubber operations, one biocontrol company, and three government-owned laboratories were, and currently one still is, involved in their commercialization. One species, currently referred to as Sporothrix insectorum, has been historically deployed in biocontrol applications in Brazilian rubber farms. However, L. heveae-infecting isolates have only been identified through morphological examinations; therefore, proper molecular assessments are needed for accurate identifications. Hence, DNA of six L. heveae-infecting isolates (five of which have been deployed in field applications) were extracted and sequenced. Multigene phylogeny found that both Simplicillium lanosoniveum and Cordyceps (formerly Isaria) sp. have been sprayed on rubber plantations to manage L. heveae populations, although the former is the only one currently applied. Simplicillium lanosoniveum and Cordyceps sp. have no relation whatsoever to true Sporothrix species associated with human and animal diseases. Therefore, our molecular data may encourage biocontrol companies to register mycoinsecticides targeting L. heveae. We also added unpublished historical accounts after contacting key contributors to the launching of this not so well-known biocontrol program in the 1980s.

Development, in vitro biocompatibility, and antitumor efficacy of acetic acid-modified Cordyceps sinensis polysaccharide nanoparticle drug delivery system

Docetaxel-loaded acetic acid conjugated Cordyceps sinensis polysaccharide (DTX-AA-CSP) nanoparticles were prepared through dialysis and their release rates in vitro, particle sizes, zeta potentials, drug loading capacities, and encapsulation efficiencies were characterized for the synthesis of AA-modified CSPs from traditional Chinese medicine Cordyceps sinensis (Berk.) Sacc. Then, the AA-modified CSPs were characterized by 1H-NMR and FT-IR. Furthermore, the biocompatibility of the delivery carrier (AA-CSP nanoparticles) was assessed on human umbilical vein endothelial cells. In vitro antitumor activity studies on DTX-AA-CSP nanoparticles were conducted on the human liver (HepG2) and colon cancer cells (SW480). The DTX-AA-CSP nanoparticles were spherical and had an average size of 98.91±0.29 nm and zeta potential within the −19.75±1.13 mV. The encapsulation efficiency and loading capacity were 80.95%±0.43% and 8.09%±0.04%, respectively. In vitro, DTX from the DTX-AA-CSP nanoparticles exhibited a sustained release, and the anticancer activities of DTX-AA-CSP nanoparticles against SW480 and HepG2 were significantly higher than those of marketed docetaxel injection (Taxotere®) in nearly all the tested concentrations. The AA-CSP nanoparticles showed good biocompatibility. This study provided a promising biocompatible delivery system for carrying antitumor drugs for cancer therapy

Optimization of liquid fermentation conditions for biotransformation zein by Cordyceps militaris 202 and characterization of physicochemical and functional properties of fermentative hydrolysates

Cordyceps militaris 202 is a potential fungus for biotransformation zein, due to its various proteases, high tolerance and viability in nature. In this article, single factor experiment and response surface methodology were applied to optimize the liquid fermentation conditions and improve the ability of biotransformation zein. The optimized fermentation conditions were as follows: inoculum concentration of 19%, volume of liquor of 130 mL/500 mL and pH of 4.7. Under this condition, the degree of hydrolysis (DH) was 27.31%. The zein hydrolysates from fungi fermentation maintained a high thermal stability. Compared to the original zein, the zein hydrolysates were found to have high solubility, which most likely results in improved foaming and emulsifying properties. Overall, this research demonstrates that hydrolysis of zein by C. militaris 202 is a potential method for improving the functional properties of zein, and the zein hydrolysates can be used as functional ingredients with an increased antioxidant effect in both food and non-food applications.(AU)

Optimization of liquid fermentation conditions for biotransformation zein by Cordyceps militaris 202 and characterization of physicochemical and functional properties of fermentative hydrolysates

Abstract Cordyceps militaris 202 is a potential fungus for biotransformation zein, due to its various proteases, high tolerance and viability in nature. In this article, single factor experiment and response surface methodology were applied to optimize the liquid fermentation conditions and improve the ability of biotransformation zein. The optimized fermentation conditions were as follows: inoculum concentration of 19%, volume of liquor of 130 mL/500 mL and pH of 4.7. Under this condition, the degree of hydrolysis (DH) was 27.31%. The zein hydrolysates from fungi fermentation maintained a high thermal stability. Compared to the original zein, the zein hydrolysates were found to have high solubility, which most likely results in improved foaming and emulsifying properties. Overall, this research demonstrates that hydrolysis of zein by C. militaris 202 is a potential method for improving the functional properties of zein, and the zein hydrolysates can be used as functional ingredients with an increased antioxidant effect in both food and non-food applications.

Genetic diversity of the entomopathogenic fungus Cordyceps tenuipes in forests and butterfly gardens in Quindío, Colombia.

Cordyceps tenuipes is an entomopathogenic fungus that infects mostly pupae of several lepidopteran families. In Colombia the species has been reported in non-disturbed tropical rain forests and more recently in butterfly gardens. The aim of this study was to assess the genetic diversity in populations of C. tenuipes present in natural (forests) and artificial (e.g. butterfly gardens) environments in the department of Quindío, Colombia, using three molecular nuclear markers ITS, TEF-1α and RPB1. All the samples evaluated corresponded morphologically and phylogenetically to C. tenuipes. The butterfly garden of Quindio Botanical Garden (QBG) showed the highest genetic diversity among all sampling localities and was very similar to that of its adjacent forest. The Amaranta Butterfly Garden (ABG), located north of QBG, showed lower genetic diversity as well as little genetic differentiation with QBG, consistent with the hypothesis of a pathogen transfer from QBG to ABG. Higher FST values were observed for TEF-1α and ITS, revealing genetic differentiation between all demes and the southern forest population. Our research constitutes the first study of the intraspecific diversity of C. tenuipes in Colombia and can serve as the first step in identifying diversity reservoirs and management of epizootic episodes caused by this fungal species.

Optimization of liquid fermentation conditions for biotransformation zein by Cordyceps militaris 202 and characterization of physicochemical and functional properties of fermentative hydrolysates.

Cordyceps militaris 202 is a potential fungus for biotransformation zein, due to its various proteases, high tolerance and viability in nature. In this article, single factor experiment and response surface methodology were applied to optimize the liquid fermentation conditions and improve the ability of biotransformation zein. The optimized fermentation conditions were as follows: inoculum concentration of 19%, volume of liquor of 130mL/500mL and pH of 4.7. Under this condition, the degree of hydrolysis (DH) was 27.31%. The zein hydrolysates from fungi fermentation maintained a high thermal stability. Compared to the original zein, the zein hydrolysates were found to have high solubility, which most likely results in improved foaming and emulsifying properties. Overall, this research demonstrates that hydrolysis of zein by C. militaris 202 is a potential method for improving the functional properties of zein, and the zein hydrolysates can be used as functional ingredients with an increased antioxidant effect in both food and non-food applications.

Evidence for Widespread Associations between Neotropical Hymenopteran Insects and Actinobacteria.

The evolutionary success of hymenopteran insects has been associated with complex physiological and behavioral defense mechanisms against pathogens and parasites. Among these strategies are symbiotic associations between Hymenoptera and antibiotic-producing Actinobacteria, which provide protection to insect hosts. Herein, we examine associations between culturable Actinobacteria and 29 species of tropical hymenopteran insects that span five families, including Apidae (bees), Vespidae (wasps), and Formicidae (ants). In total, 197 Actinobacteria isolates were obtained from 22 of the 29 different insect species sampled. Through 16S rRNA gene sequences of 161 isolates, we show that 91% of the symbionts correspond to members of the genus Streptomyces with less common isolates belonging to Pseudonocardia and Amycolatopsis. Electron microscopy revealed the presence of filamentous bacteria with Streptomyces morphology in brood chambers of two different species of the eusocial wasps. Four fungal strains in the family Ophiocordycipitacea (Hypocreales) known to be specialized insect parasites were also isolated. Bioassay challenges between the Actinobacteria and their possible targeted pathogenic antagonist (both obtained from the same insect at the genus or species level) provide evidence that different Actinobacteria isolates produced antifungal activity, supporting the hypothesis of a defensive association between the insects and these microbe species. Finally, phylogenetic analysis of 16S rRNA and gyrB demonstrate the presence of five Streptomyces lineages associated with a broad range of insect species. Particularly our Clade I is of much interest as it is composed of one 16S rRNA phylotype repeatedly isolated from different insect groups in our sample. This phylotype corresponds to a previously described lineage of host-associated Streptomyces. These results suggest Streptomyces Clade I is a Hymenoptera host-associated lineage spanning several new insect taxa and ranging from the American temperate to the Neotropical region. Our work thus provides important insights into the widespread distribution of Actinobacteria and hymenopteran insects associations, while also pointing at novel resources that could be targeted for the discovery of active natural products with great potential in medical and biotechnological applications.

Cordyceps sinensis promotes immune regulation and enhances bacteriostatic activity of PA-824 via IL-10 in Mycobacterium tuberculosis disease

PA-824 is a novel bicyclic nitroimidazole anti-tuberculosis (TB) drug. Cordyceps sinensis (Berk.) Sacc. (CS) was proven to be a good immunomodulatory compound. This research aimed to investigate the effect of CS on PA-824 in Mycobacterium tuberculosis (M.tb) infected mice (female CBA/J mice, 6 to 8 weeks of age and 20±2 g of weight). Mice were randomly assigned to 4 groups: PA-824, CS, PA-824+CS, and control. To verify the effect of PA-824 and CS on M.tb, after drug administration, mice lungs were harvested and bacterial colony formations were measured. Cells were isolated from infected lungs and spleens to analyze the percentage of CD4+ T cells (CD11a positive). Lung cells were cultured to detect the secretion of interferon-γ (IFN-γ) and interleukin-10 (IL-10) by ELISA. IFN-γ and IL-10 double-positive CD4+ cells in peripheral blood were measured by flow cytometry. The expression levels of IL-2 and IL-10 in mice lungs were analyzed by real-time PCR and western blot. Results showed that PA-824 combined with CS led to the lowest lung colony-forming units (CFU) counts among treated groups. Furthermore, this beneficial outcome might be associated with the decreased CD11a on CD4+ cells in mice lungs and spleens. Moreover, the suppressed secretion of IFN-γ and IL-10, and IL-10 expressions, as well as the decreased IFN-γ and IL-10 double-positive CD4+ cells in blood, could also be associated with the positive effect. However, no significant effect on IL-2 production was found. The combination of PA-824 and CS had more effective bacteriostatic and immunomodulatory effects on M.tb infected mice than PA-824 alone. In conclusion, CS has the potential to be an effective adjuvant in TB treatment.

Characterizations of a new Cordyceps cicadae isolate and production of adenosine and cordycepin

Cordyceps is a fastidious pathogenic fungus infecting insects, and recent years have witnessed rapid progress in its medical properties. In this study, a wild isolate, C. cicadae MP12, was characterized through in vitro cultivation and its nuclear small-subunit (SSU) ribosomal DNA (rDNA) data. In vitro culture of C. cicadae MP12 was established by growing its fruiting bodies in a solid matrix. C. cicadae MP12 was inoculated into Cryptotympana atrata cicada pupae for in vivo culture, where the fungi developed its fruiting body as well. The contents of adenosine and cordycepin in dried fruiting bodies after culture were 1421.45µg/g and 1398.12 µg/g, respectively. Therefore, the established cultures from this study could be used for the production of various medically important metabolic substances.

Characterizations of a new Cordyceps cicadae isolate and production of adenosine and cordycepin.

Cordyceps is a fastidious pathogenic fungus infecting insects, and recent years have witnessed rapid progress in its medical properties. In this study, a wild isolate, C. cicadae MP12, was characterized through in vitro cultivation and its nuclear small-subunit (SSU) ribosomal DNA (rDNA) data. In vitro culture of C. cicadae MP12 was established by growing its fruiting bodies in a solid matrix. C. cicadae MP12 was inoculated into Cryptotympana atrata cicada pupae for in vivo culture, where the fungi developed its fruiting body as well. The contents of adenosine and cordycepin in dried fruiting bodies after culture were 1421.45µg/g and 1398.12 µg/g, respectively. Therefore, the established cultures from this study could be used for the production of various medically important metabolic substances.

Characterizations of a new Cordyceps cicadae isolate and production of adenosine and cordycepin

Cordyceps is a fastidious pathogenic fungus infecting insects, and recent years have witnessed rapid progress in its medical properties. In this study, a wild isolate, C. cicadae MP12, was characterized through in vitro cultivation and its nuclear small-subunit (SSU) ribosomal DNA (rDNA) data. In vitro culture of C. cicadae MP12 was established by growing its fruiting bodies in a solid matrix. C. cicadae MP12 was inoculated into Cryptotympana atrata cicada pupae for in vivo culture, where the fungi developed its fruiting body as well. The contents of adenosine and cordycepin in dried fruiting bodies after culture were 1421.45µg/g and 1398.12 µg/g, respectively. Therefore, the established cultures from this study could be used for the production of various medically important metabolic substances.