Filamentous fungi with high paraquat-degrading activity isolated from contaminated agricultural soils in northern Thailand.

The contamination of paraquat (1,1'-dimethyl-4,4'-bipyridylium dichloride) herbicide from the farming area has become a public concern in many countries. This herbicide harms to human health and negatively effects the soil fertility. Several methods have been introduced for the remediation of paraquat. In this study, 20 isolates of the paraquat-tolerant fungi were isolated from the contaminated soil samples in northern Thailand. We found that isolate PRPY-2 and PFCM-1 exhibited the highest degradation activity of paraquat on synthetic liquid medium. About 80 and 68% of paraquat were removed by PRPY-2 and PFCM-1 respectively after 15 days of cultivation. Based on the morphological characteristic and molecular analysis, the fungal isolate PRPY-2 and PFCM-1 were identified as Aspergillus tamarii and Cunninghamella sp. respectively. The biosorption of paraquat on these fungal mycelia was also investigated. It was found that only 8-10% of paraquat could be detected on their mycelia, while 24-46% of paraquat was degraded by fungal mycelia. This is the first report on paraquat degrading ability by A. tamarii and Cunninghamella sp. It is demonstrated that these filamentous fungi are promising microorganisms available for remediation of paraquat contaminated environment.

Use of Bacillus subtilis isolates from Tua-nao towards nutritional improvement of soya bean hull for monogastric feed application.

UNLABELLED: Soya bean hull (SBH) is a cheap and high-fibre content feed ingredient that obtained after soya bean oil extraction. Microbial fermentation was expected to improve SBH qualities before applying to animals, especially monogastric animals. Two bacterial strains, Bacillus subtilis MR10 and TK8 that were isolated from Tua-nao, a traditional fermented soya bean in northern Thailand, were used for fermented soya bean hull (FSBH) production. Both could easily grow at 37°C in SBH as the sole substrate. MR10 produced the highest ß-mannanase activity (400 U g(-1) SBH) on day 2, while TK8 produced the highest cellulase activity (14·5 U g(-1) SBH) on day 3. After fermentation, the nutritional quality of SBH was obviously improved by an increase in soluble sugars, soluble proteins, crude protein and crude lipid, and a decrease in the content of raffinose family oligosaccharides. Scavenging activity (%) of SBH against ABTS radical cation was also increased from 14 to 27 and 20% by MR10 and TK8 fermentation, respectively. According to the GRAS property of these both strains and various improvements of nutritional values, the fermented SBH proved to be a potential feed ingredient, especially for the monogastric animals. SIGNIFICANCE AND IMPACT OF THE STUDY: Normally, soya bean hull has been recognized as only a worthless by-product from soya bean oil production process because of its low utilizable nutrients. Our study introduced an alternative way to utilize this worthless residue using biotechnological knowledge. The nutritional quality of soya bean hull was improved by microbial fermentation. Fermented soya bean hull can be used as a cheap, safe and high-nutrient feed ingredient for livestock production, especially monogastric animals, to promote their growth performances, instead of using antibiotics in some regions of the world.

Screening and selection of Bacillus spp. for fermented corticate soybean meal production.

AIMS: To screen and select the Bacillus spp. from Tua-nao of northern Thailand for fermented corticate soybean meal (FCSBM) production. METHODS AND RESULTS: After isolation of Bacillus spp. from Tua-nao was carried out, cellulase, hemicellulases (i.e., ß-mannanase and xylanase) and phytase production by isolated Bacillus spp. were determined. B. subtilis isolate MR10 showed the highest ß-mannanase, xylanase and phytase production at 280, 41 and 16 U g(-1) substrate, respectively, while the highest cellulase production was found in TK8 at 25 U g(-1) substrate. FCSBMs produced by single starter and mixed starter of both isolates showed the better properties than those of corticate soybean meal (CSBM), i.e., higher in soluble sugar, protein and phosphate content, smaller sugar molecules and better digestibility and absorbability than those of CSBM. Moreover, FCSBMs had no toxicity effect on mouse fibroblast cell line (3T3) but had an inhibitory effect on lung cancer cell line (CorL23). CONCLUSIONS: B. subtilis isolate MR10 and TK8 were selected for FCSBMs production because of their role as nutritional enhancer for CSBM and their safety. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study were useful for FCSBM production process that can be applied as feed ingredient for monogastric animals.

Response surface method for polyhydroxybutyrate (PHB) bioplastic accumulation in Bacillus drentensis BP17 using pineapple peel.

Polyhydroxybutyrate (PHB) is a biodegradable biopolymer which is useful for various applications including packing, medical and coating materials. An endospore-forming bacterium (strain BP17) was isolated from composted soil and evaluated for PHB production. Strain BP17, taxonomically identified as Bacillus drentensis, showed enhanced PHB accumulation and was selected for further studies. To achieve maximum PHB production, the culture conditions for B. drentensis BP17 were optimized through response surface methodology (RSM) employing central composite rotatable design (CCRD). The final optimum fermentation conditions included: pineapple peel solution, 11.5% (v/v); tryptic soy broth (TSB), 60 g/L; pH, 6.0; inoculum size, 10% (v/v) and temperature, 28°C for 36 h. This optimization yielded 5.55 g/L of PHB compared to the non-optimized condition (0.17 g/L). PHB accumulated by B. drentensis BP17 had a polydispersity value of 1.59 and an average molecular weight of 1.15x105 Da. Thermal analyses revealed that PHB existed as a thermally stable semi-crystalline polymer, exhibiting a thermal degradation temperature of 228°C, a melting temperature of 172°C and an apparent melting enthalpy of fusion of 83.69 J/g. It is evident that B. drentensis strain BP17 is a promising bacterium candidate for PHB production using agricultural waste, such as pineapple peel as a low-cost alternative carbon source for PHB production.

Bioactive compounds content and their biological properties of acetone extract of Cuscuta reflexa Roxb. grown on various host plants.

The present study is aimed to evaluate the total phenolic and flavonoid contents, and free phenolic compounds of acetone extract of Cuscuta reflexa grown on five different hosts: Coccinia grandis, Ficus racemosa, Samanea saman, Streblus asper and Zollingeria dongnaiensis, and to explore the antioxidant activities, α-glucosidase and tyrosinase inhibitory properties of the extracts. The highest level of total phenolic and flavonoid contents were observed in the extract of C. reflexa that was grown on S. asper (65.45 mg GAE/g extract) and C. grandis (97.83 mg QE/g extract), respectively. According to HPLC results, vanillic acid, rutin and quercetin were found in all extracts of C. reflexa grown on diversified hosts. The extract of C. reflexa grown on C. grandis possessed the greatest antioxidant activities (DPPH; 251.64 µg/mL, FRAP; 26.44 mg GAE/g extract), α-glucosidase inhibition accounted for 84.36 per cent and antityrosinase activity was at 18.29 mg KAE/g sample.

Molecular and morphological characterization of Pyricularia and allied genera.

The phylogenetic relationships of Pyricularia species and species from related genera were established from sequences of the internal transcribed spacer ribosomal RNA gene. Phylogenetic analysis disclosed a consistent correlation with spore morphology. Most Pyricularia species studied, and two species of Dactylaria that have obpyriform conidia, fell within the Magnaporthaceae cluster with high bootstrap support. Pyricularia variabilis was more related to Dactylaria, Tumularia or Ochroconis species than to the Magnaporthaceae. Dactylaria and species of Nakataea, Ochroconis, Pyriculariopsis and Tumularia were distinct from the Magnaporthaceae, and the genus Dactylaria is polyphyletic. The combination of morphological and molecular characters, such as spore morphology and ITS ribosomal DNA sequences data, suggested that conidial shape could be a primary character to distinguish Pyricularia from related genera.

Microfungi on the Pandanaceae: Linocarpon lammiae sp. nov., L. siamensis sp. nov. and L. suthepensis sp. nov. are described with a key to Linocarpon species from the Pandanaceae.

Linocarpon species are reported from Pandanaceae in Australia, Brunei, Hong Kong, Nepal, New Zealand, Philippines, Seychelles, Thailand and Vanuatu. Linocarpon lammiae sp. nov. were collected on decaying leaves of Pandanus tectorius in Hong Kong. Linocarpon siamensis sp. nov. and L. suthepensis sp. nov. were collected from decaying leaves of P. penetrans in Thailand. These taxa are described, illustrated and compared with Linocarpon species with similar ascospore morphology and dimensions. Included are a synoptic table, which compares the new species to similar known species, and a dichotomous key to species of Linocarpon known from members of the Pandanaceae.

L-malic acid production by an albino strain of Monascus araneosus.

An albino mutant was isolated after treating Monascus araneosus AHU9087 with N-methyl-N'-nitro-N-nitrosoguanidine. All other physiological and biochemical characteristics were retained. The mutant did not produce any pigment but produced L-malic acid at 28 g/l, compared with 20 g/l by the parent strain, in media containing 10% (w/v) glucose after incubation under aerobic conditions for 5 days at 37°C.

Thermus 2S from Thai hot springs: isolation and immobilization.

Three strains of thermophilic bacteria producing extracellular protease have been isolated from hot springs in Chiang Mai, Thailand. The bacterium producing the highest amount of protease has been selected and identified as belonging to the genusThermus, and is tentatively calledThermus 2S. The isolate is a Gram-negative, rod shaped bacterium. It exhibited maximum growth around 60°C at pH 7. Entrapment of the microbial cells in calcium alginate maintained the cell viability. Protease production from immobilized cells using 2 g wet cells per 10 ml 3% (w/v) sodium alginate was higher than that from a free-cell system using 2% inoculum.

Endophytic fungi from Amomum siamense.

Endophytic fungi were isolated from apparently healthy organs of the wild ginger Amomum siamense Criab.. including leaves, pseudostems, and rhizomes, collected from two sites in Doi Suthep-Pui National Park, Thailand. Endophytes were relatively common with an isolate prevalence of 70%-83% at the two sites sampled in the wet and dry seasons. The endophyte assemblages from the two sites were diverse and comprised 7 Ascomycetes and 26 mitosporic fungi. Colletotrichum "gloeosporioides" (Penz.) Penz. & Sacc. in Penz., Glomerella spp., xylariaceous fungi, and Phomopsis spp. were consistently dominant as endophytes on Amomum siamense, but many rare species and mycelia sterilia were also recorded. There was no significant difference between the number of isolates recovered from leaves containing vein tissues and those containing intervein tissues, independent of leaf age. Most taxa showed a preference for either leaf tissue or pseudostems. Two new Ascomycetes species. Gaeumannomyces amomi and Leiosphaerella amomi, were discovered from leaves and rhizomes, respectively, and four species of Pyricularia, including three new species, were isolated from leaves.