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Isolation and Characterization of Airborne Mushroom Damaging Trichoderma spp. from Indoor Air of Cultivation Houses Used for Oak Wood Mushroom Production Using Sawdust Media

  • Kim, Jun Young;Kwon, Hyuk Woo;Lee, Dong Hyeung;Ko, Han Kyu;Kim, Seong Hwan
    • The Plant Pathology Journal
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    • v.35 no.6
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    • pp.674-683
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    • 2019
  • Some species of the Trichoderma genus are reported as the major problem in oak wood mushroom production in Korea. In spite of economic loss by the fungi, scientific information on airborne Trichoderma species is not much available. To generate information for disease management development we analyzed airborne Trichoderma. A total of 1,063 fungal isolates were purely obtained from indoor air sampling of cultivation houses used for oak wood mushroom using sawdust media. Among the obtained isolates, 248 isolates were identified as Trichoderma fungi including T. harzianum, T. atroviride, T. citrinoviride, and T. pseudokoningii, by morphological and molecular analysis. T. harzianum was dominant among the four identified species. All the four Trichoderma species grew fast on solid nutrient media tested (potato dextrose agar [PDA], malt extract agar [MEA], Czapek's Dox + yeast extract agar [CYA] and cornmeal dextrose agar). Compact mycelia growth and mass spore production were better on PDA and CYA. In addition, T. harzianum and T. citrinoviride formed greenish and yellowish mycelium and spores on PDA and CYA. Greenish and yellowish pigment was saturated into PDA only by T. pseudokoningii. These four Trichoderma species could produce extracellular enzymes of sawdust substrate degradation such as β-glucosidase, avicelase, CM-cellulase, amylase, pectinase, xylanase, and protease. Their mycelia inhibited the growth of oak wood mushroom mycelia of two tested cultivars on dual culture assay. Among of eleven antifungal agents tested, benomyl was the best to inhibit the growth of the four Trichoderma species. Our results demonstrate that the airborne Trichoderma fungi need to be properly managed in the cultivation houses for safe mushroom production.

Studies on the Mycelial Production of Aspergillus niger with Use of Cassava Flour Starch (Cassava 전분(澱粉)을 이용한 검정곰팡이(Aspergillus niger)의 균체생산(菌體生産)에 관한 연구(硏究))

  • Bang, Jeong-Hee;Kim, Jong-Hyup
    • The Korean Journal of Mycology
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    • v.15 no.4
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    • pp.231-237
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    • 1987
  • A. niger IMI 41873 was shake-cultured in Cassava starch medium, then mycelial dry weight and mycelial protein were measured. The effects of Cassava starch concentrations, various nitrogen sources and concentrations on the levels of mycelial production and mycelial protein were investigated. The results were as follows; In each medium containg 4%, 6%, 8% and 10% of Cassava starch, mycelial production was 13.35 mg/ml, the medium containg 6% of Cassava starch was proved to be most effective. The levels of mycelial protein in shake culture with medium of 6% Cassava starch and $NaNO_3(1.5\;g/l)$ were the highest. Among nitrate, ammonium sulfate, and urea, the nitrate was the most effective on mycelial production.

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Optimal Conditions for the Production of Salt-tolerant Protease from Aspergillus sp. 101 and Its Characteristics (Aspergillus sp. 101로부터 내염성 단백분해효소 생산을 위한 최적 조건 및 특성)

  • Hwang, Joo-Yeon;Choi, Seung-Hwa;Lee, Si-Kyung;Kim, Sang-Moo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.11
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    • pp.1612-1617
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    • 2009
  • Aspergillus sp. 101 was isolated from the Korean traditional soybean paste for the production of a salt-tolerant protease. The optimal condition for the production of a salt-tolerant protease was determined with various energy sources such as carbon, nitrogen, and protein, and at different culture conditions such as temperature, pH, incubation time and NaCl concentration. The most favorable organic nitrogen sources were 2% defatted soybean flour (DSF) and soy protein isolate (SPI). Optimal pH and temperature were pH 6.0 and $25{\sim}27^{\circ}C$, respectively. Therefore, Aspergillus sp. 101 protease was a mild acid (or neutral) protease. Protease production was the highest at 0.1% concentration of $CaCO_3,\;K_2HPO_4$ and Arabicgum. Aspergillus sp. 101 could grow in culture medium at 15% NaCl concentration and produce a salt-tolerant protease even at 7% NaCl. The cell mass and protease activity of Aspergillus sp. 101 cultured in a modified medium was comparatively higher in Czapek dox and protease producing media. Hence, Aspergillus sp. 101 protease can be utilized in soy or fish sauce industry as a salt-tolerant protease starter.

Salvage Chemotherapy in Recurrent Platinum-Resistant or Refractory Epithelial Ovarian Cancer with Carboplatin and Distearoylphosphatidylcholine Pegylated Liposomal Doxorubicin (Lipo-Dox®)

  • Khemapech, Nipon;Oranratanaphan, S.;Termrungruanglert, W.;Lertkhachonsuk, R.;Vasurattana, A.
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.3
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    • pp.2131-2135
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    • 2013
  • Background: To evaluate the efficacy and safety of distearoylphosphatidylcholine pegylated liposomal doxorubicin (DPLD) combined with carboplatin for the treatment of platinum resistant or refractory epithelial ovarian cancer (EOC) or fallopian tube cancer. Materials and Methods: A retrospective analysis of women who received DPLD with carboplatin for recurrent EOC or fallopian tube cancer in King Chulalongkorn Memorial Hospital Thailand from January 2006 to August 2011 was conducted. Patients were identified from the medical records and data on demographic factors, stage, histology, surgical findings, cytoreduction status, and prior chemotherapies were abstracted. The efficacy and toxicity of DPLD/carboplatin were evaluated. Progression-free (PFS) and overall survival (OS) were estimated by the Kaplan-Meier method. Results: A total of 65 patients, 64 with platinum resistant or refractory epithelial ovarian cancer and 1 with fallopian tube cancer, were enrolled. DPLD and carboplatin were given for an average of 4.46 cycles per patient with a total of 273 cycles. Among the 65 evaluable patients, 0% achieved CR, 7.69% PR, 15.4% SD and 76.% PD. The overall response rate was 23.1%. With a median follow-up of 27.4 months, the median progression-free and median overall survival in the 36 patients was 4.46 months and 8.76 months respectively. In the aspect of side effects, palmar-plantar erythrodysesthesia (PPE) occurred in 33.3% (Grade I 22.2%, Grade II 11.1%) and mucositis in 41.7% (Grade I 27.8%, Grade II 13.9%) of all treatment cycles, all Grade 1 or 2. Anemia, leukopenia and thrombocytopenia occurred in 58.3% (Grade I 41.7%, Grade II 16.7%), 66.7% (Grade I 47.2%, Grade II 19.4%), and 22.2% (Grade I 16.6%, Grade II 5.56%) of cycle respectively, and were mostly Grade 1 or 2. Conclusions: DPLD, the second-generation PLD drug combined with carboplatin every 4 weeks, is effective and has low toxicity for treatment of patients with recurrent platinum-resistant or refractory epithelial ovarian cancer.

Comparative Outcome of Thai Pediatric Osteosarcoma Treated with Two Protocols: the Role of High-Dose Methotrexate (HDMTX) in a Single Institute Experience

  • Choeyprasert, Worawut;Pakakasama, Samart;Sirachainan, Nongnuch;Songdej, Duantida;Chuansumrit, Ampaiwan;Anurathapan, Usanarat;Hongeng, Suradej;Nartthanarung, Adisak
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.22
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    • pp.9823-9829
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    • 2014
  • Background: High-dose methotrexate (HD-MTX) is recognized as an efficient component of therapy against pediatric osteosarcoma in combination with other drugs such as cisplatin (CDP), carboplatin (CBDCA), doxorubicin (ADM), etoposide (VP-16) and ifosfamide (IFO). Objectives: To demonstrate the feasibility and effectiveness of the HD-MTX/CDP/DOX/VP-16/IFO [MTX(+)] protocol comparable to CDP/ADM/CBDCA/IFO [MTX(-)] for treating childhood osteosarcoma at Ramathibodi Hospital (1999-2014). Materials and Methods: A retrospective analysis was conducted of osteosarcoma patients aged less than 18 years treated with two chemotherapeutic regimens between 1999 and 2014. A total of 45 patients received the MTX(-) and 21 the MTX(+) protocol. Results: Overall limb-salvage and amputation rate were 12.9% and 77.7%, respectively. Kaplan-Meier analysis results for 3-year disease free survival (DFS) and overall survival (OS) regardless of treatment regimens were $43.4{\pm}6.0%$ and $53.2{\pm}6.1%$ respectively. The 3-year DFS and OS were improved significantly with the MTX(+) protocol compared to MTX(-) protocol (p=0.010 and p=0.009, log rank test) [$69.8{\pm}10.5%$, $79.8{\pm}9.1%$ for MTX(+) and $31.1{\pm}6.9%$, $42.2{\pm}7.4%$ for MTX(-) protocol, respectively]. Patients with metastatic osteosarcoma treated with the MTX(+) protocol had statistically significant higher 3-year DFS and OS than those treated with the MTX(-) protocol ($66.7{\pm}13.6%$ and $15.0{\pm}8.0%$ for 3-year DFS, p=0.010, $73.3{\pm}13.2%$ and $20{\pm}8.9%$ for 3-year OS, p=0.006, respectively). The independent risk factors for having inferior 3-year DFS and OS were poor histological response (tumor necrosis <90%) and treatment with the MTX(-) protocol. The multivariate analysis identified only the treatment with the MTX(-) protocol as an independent predictor of inferior OS with a hazard ratio (HR) of 3.53 (95% confidence interval of 1.2-10.41, p=0.022). Conclusions: Our study demonstrated the tolerability, feasibility and efficacy of the HDMTX-based regimen improving the survival rate in pediatric osteosarcoma cases, in line with reports from developed countries.

Studies on the Varietal Resistance and Effects of Nutrients for Fungal Growth of Pepper Anthracnose Disease Caused by Colletotrichum dematium f. sp. capsicum (고추 탄달병(炭疸病)에 대한 품종(品種) 저항성(抵抗性) 및 병원균(病原菌) 생장(生長)에 미치는 영양원(營養源)의 효과(效果))

  • Chang, Sun-Hwa;Chung, Bong-Koo
    • The Korean Journal of Mycology
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    • v.13 no.4
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    • pp.227-233
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    • 1985
  • Studies on the varietal resistance and effects of nutrients for fungal growth were carried out in order to obtain basic materials for breeding resistant variety and control measures of the red pepper anthracnose disease caused by Colletotrichum dematium f. sp. capsicum. Four cultivars such as Kumchang No. 2, Bulamhouse, Pakistan, Hongilpum were resistant among twenty-one pepper cultivars, and five cultivars including Taiwan pepper were moderate. The remaining twelve cultivars including H-038 and Saegochu were susceptible. These susceptible cultivars were mostly belonged to sweet taste cultivars. Glucose was known the best source for fungal growth, and near 3 percent of carbon concentration was the best for mycelial growth of the fungus. Conidial sporulation was rather decreased by adding high concentrations of C-source, whereas fungal dry weight was a positive tendency in proportion to increasing carbon concentrations. N-sources and vitamins were not remarkable as that for carbon, and rather a decreasing trend for mycelial growth by adding N-source. Especially, the lowest of mycelial growth was in the case of Czapek-Dox plus ammonium sulfate. The medium plus vitamins either niacin or thiamine was slightly increased to mycelial growth.

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Composition of a New Medium for Mycelial Growth of Hericium erinaceus (노루궁뎅이버섯(Hericium erinaceus)의 새로운 균사배양기의 조성)

  • Ko, Han-Gyu;Kim, Dong-Myong;Park, Won-Mok
    • The Korean Journal of Mycology
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    • v.25 no.4 s.83
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    • pp.369-376
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    • 1997
  • These researches were carried out for improvement of medium for mycelial growth of Hericium erinaceus isolate KU-1. It grew well at pH 4 and $25^{\circ}C$. Glucose and sucrose were favorable carbon sources for mycelial growth. As nitrogen sources, ammonium acetate and arginine enhanced mycelial growth. Optimum C/N ratio was 200. Based On the results, the following recipe is suggested for synthetic medium for the mycelial growth: glucose 18.02 g, arginine 2.613 g, ammonium acetate 2.313 g, $CaCl_2\;0.33\;g$, $KH_2PO_4\;8.5\;g$, $MgSO_4{\cdot}7H_2O\;2.0\;g$, $FeSO_4{\cdot}7H_2O\;0.02\;g$, $ZnSO_4{cdot}7H_2O\;0.02\;g$, $MnSO_4{\cdot}7H_2O\;0.02\;g$, water 1 liter. This medium was superior for the mycelial growth to other conventional media such as Yeast malt extract agar (YMA), Park medium, Potato dextrose agar (PDA), Malt extract agar, Czapek-dox agar, Macaya-lizano medium and Yeast extract agar. This new synthetic medium is designated as Ko medium.

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Physical Characteristics and Antagonistic Effect of Ampelomyces (Ampelomyces의 생리적 특성 및 길항능력)

  • Kim, Ji-Young;Lee, Wang-Hyu;Kim, Hyung-Moo
    • Research in Plant Disease
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    • v.15 no.3
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    • pp.209-216
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    • 2009
  • During the period of June, 2005 to May, 2008, 44 host plants infected with powdery mildew were collected in the Jeon-ju and Jang-su districts of Jeonbuk province and in the Jang-sung district of Jeonnam province. The hyperparasites, Ampelomyces were confirmed in 12 plant species. Most of the pycnidium shapes of Ampelomyces were circular or oval shaped, and the sizes were different even within the same host plant, and also the color of pycnidium was ranged from light brown to dark brown. Ampelomyces species were isolated from 4 hosts including Impatiens balsamina L., Cucurbita pepo, Rudbeckia laciniata var. elatier and Youngia sonchifolia, and thus the most appropriate 12 Ampelomyces strains for the current experiment were selected. When analyzing the selected 12 strains' incubational and nutritional characteristics, the malt extract agar was the most appropriate media. When investigating the effect of osmotic pressure on the spore germination, 0.15M NaCl concentration was the optimum germination concentration. When the isolated Ampelomyces sp. was tested in-vitro, it was found to be effective to control in other plant pathogens, isolated Ampelomyces showed no pathogenicity to the plant. strains isolated . studied on rDNA ITS sequence analysis. The rDNA ITS sequence data of Ampelomyces sp. isolate BSLAH16 from Impatiens balsamina L. were analyzed and identified.

Studies on the Ecology of Occurrence and Identification of Typhula Snow Mold of Graminous Plants -II. Several Factors Affecting Growth of Typhula incarnata- (화본과식물에 발생하는 설부소입균핵병균(雪腐小粒菌核病菌)의 동정 및 발생상태에 관한 연구 -II. Typhula incarnata의 생육에 미치는 몇 가지 요인-)

  • Kim, Jin-Won;Lee, Du-Hyung;Shim, Gyu-Yul
    • The Korean Journal of Mycology
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    • v.20 no.1
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    • pp.37-43
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    • 1992
  • Typhula incarnata grew over a temperature range of -5 to $20^{\circ}C$ with maximum growth at 10 to $15^{\circ}C$. Sclerotial production for T. incarnata was greatest at the higher temperature. Maximum mycelial growth of this pathogen occurred from pH 5.4 to 6.2. When carbon sources were added to a basal salt medium (Czapek's dox agar) at 5 g carbon sources/l, inulin, soluble starch, galactose, glucose, mannose, manitol, sucrose, maltose, cellobirose, trehalose, raffinose, and dextrin supported growth better than other carbon sources did. Of the twenty-three nitrogen sources tested, glycine, serine, ammonium sulfate, asparagine, asparatic acid, and ${\beta}-alanine$ were the most favorable for mycelial growth of T. incarnata. Cystine and cysteine were poor nitrogen sources. Ammonium salt of nitrogen sources supported growth better than nitrate salt of nitrogen sources. Potato dextrose agar, oat meal agar, and V-8 juice agar were the most favorable for mycelial growth and sclerotial formation. Appropriate addition of pepton to PDA decreased mycelial dry weight, but sucrose supported good growth of T. incarnata. Percent viable sclerotia of T. incarnate buried in bentgrass soil decreased from 2 months after treatment remarkably. Trichoderma riride and bacteria were isolated from non-germinated sclerotia. Live orchard grass leaf pieces within the soil were colonized by T. incarnata better than sterile and unsterile dead leaf pieces at $0^{\circ}C$. Saprophytic ability of T. incarnate on sterile leaf sheath occurred better at $0^{\circ}C$ than at $10^{\circ}C$. Saprophytic microflora consisting of Cladosporium sp., Fusarium sp., Mucor sp., Pythium sp., and unidentified fungi were the competitors for the sterilized and unsterilized substrate, but their colonization was not find on live leaf sheath buried in the soil at $0^{\circ}C$. In the effect of fungicides to Typhula snow mold disease of creeping bentgrass, mixture of polyoxin and thiram was the most effective, followed by iprodione, mixture of iprodione and oxine copper, thiophanate-methyl, myclobutanil, and tolclofos-methyl.

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Influence of Nutritional Supplementation to the Substrate on Vegetative and Reproductive Growth of Winter Mushroom, Flammulina velutipes (Curt. ex Fr.) Sing. and Chemical Changes of the substrates Produced during Growth of the Fungus (톱밥 배지(培地)에 대(對)한 영양첨가(營養添加)가 팽이버섯의 생장(生長)및 배지(培地)의 화학적(化學的) 성분(成分) 변화(變化)에 미치는 영향(影響))

  • Chang, Hak-Gil
    • The Korean Journal of Mycology
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    • v.4 no.1
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    • pp.31-44
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    • 1976
  • The studies were carried out to examine the effects of supplementation of nutritional substances and physical conditions in substrate on the mycelial growth and yield of fresh sporophores of winter mushroom, Flammulina velutipes(Curt. ex Fr.) Sing. and to obtain further informations on the nutritional requirements of the fungus with reference to improvement of substrate through [analysis of chemical composition of the substrates during the cultivation period. The results obtained are summarized as follows: 1. The best yield of fresh sporophores, 84.4 g per 280 g substrate in a bottle, was obtained from the mixture of poplar sawdust 10 and rice bran 3 by volume when Flammulina velutipes was cultivated on the poplar sawdust supplemented by rice bran, wheat bran, cattle manure and various combinations of these materials as nutrient sources. The substrates of poplar sawdust 10 plus rice bran 3 and 2 or wheat bran 3 with a higher yield of fresh sporophores showed a comparatively higher content of total nitrogen. total sugar, and potassium. 2. The mycelial growth of the fungus was compared on the substrates of poplar sawdust supplemented by the several nutrient sources and poplar sawdust alone. The fastest linear growth occurred on substrates of poplar sawdust alone and poplar sawdust plus cattle manure deficient in sugar and nitrogen sources, but mycelial density was more sparse on the substrates. Also, growth in a solution extracted from these substrates was very meager. 3. In the substrates which varied with bulk density and moisture content optimum bulk density and moisture content for mycelial growth was 0.2g/cc and 72% on a dry weight basis, respectively, but the highest yield of fresh sporophores was obtained at the bulk density of 0.3g/cc and moisture content of 67%. 4. By increasing the ratio of rice bran in poplar sawdust the loss of total weight and ash, content at each stage was increased, and during the cultivation period of 75 days, loss of total weight of the substrates at inoculation was 17.8 to 28.8% and ash content increased about 12%. 5. 11 to 14% of the cellulose and 3 to 4% of the lignin content per original substrate were decreased without a great difference depending of the mixing ratio of rice bran. The soluble glucose concentration in the substrates was increased during the same period. 6. In the process of vegetative and reproductive growth of the fungus upon the substrates, the total nitrogen was increased in quantity per dry weight of sample but was reduced in absolute quantity to a minute extent. There is no great changes in content of organic nitrogen including amino acid nitrogen, and hydrolysable ammonium nitrogen during the vegetative growth period, but occurrence of sporophores resulted in a decrease in the nitrogen content of these forms. On the one hand, by an increase of additive amounts of rice bran, nitrogen contents of these forms were higher and the reduction range during the reproductive growth period became wider. 7. Mycelial growth of the fungus was accelerated in various liquid media supplemented with organic nitrogen sources such as peptone and yeast extract in comparison with addition of inorganic nitrogen sources. Furthermore, mycelial growth was mere vigorous in the media with higher content of organic nitrogen sources.

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