The effect of gamma irradiation of microbiological and physicochemical properties of raw ingredients (thirteen kinds of protein sources) for the mixed feed were investigated. The total aerobic bacteria counts in the samples was ranged from $10^2$ to $10^7$cells/g. After 5 to 7 kGy irradiation, the total count was reduced by 3 to 4 log cycles. Coliforms were detected in seven samples and the range was from $10^4$ to $10^6$cells/g. Enteric pathogens were found only in fish meal and poultry byproduct as $10^2$ to $10^6$cells/g. They were sensitive to radiation and completely sterilized by 3 to 5 kGy irradiation. Total fungi count was ranged from $10^1$ to $10^4$cells/g in all samples. They were osmophiles such as Aspergillus, Penicillium, Cladosporium, Aureobasidium and Rhizopus and were destroyed by 3 to 7 kGy irradiation. Five species of potential mycotoxin producers including Aspergillus flavus were also identified. Physicochemical properties, such as total amino acid content, TBA value, POV and color difference showed that irradiation with optimum does was less detrimental than autoclaving.
Objective: The study was conducted to determine the effects of body weight (BW) and fiber sources on nutrient digestibility, fiber fermentation and short chain fatty acids (SCFA) concentration in different intestinal segments of growing pigs fed high-fiber diets. Methods: Nine barrows with initial BW of 25.17±0.73 kg and 9 barrows with initial BW of 63.47±2.18 kg were allotted to a duplicate 9×2 Youden Square design with 3 dietary treatments and 2 periods. The dietary treatments were formulated with 3 different high-fiber ingredients: corn bran, sugar beet pulp, and soybean hulls, respectively. Each diet was fed to 3 barrows with different stage of BW in each period. Results: There were no differences in the apparent ileal digestibility (AID) of most nutrients between pigs at different BW stages. Pigs at 60 kg had greater (p<0.05) apparent total tract digestibility (ATTD) of total dietary fiber (TDF), soluble dietary fiber (SDF) and insoluble dietary fiber (IDF), and had greater (p<0.05) hindgut disappearance of IDF and cellulose than pigs at 25 kg. The acetate, propionate and total SCFA concentrations in ileal digesta and feces of pigs at 60 kg were greater (p<0.05) than those of pigs at 25 kg. In addition, fiber sources affected (p<0.05) the AID of gross energy (GE), organic matter (OM), ether extract (EE), crude protein, SDF and hemicellulose, the hindgut disappearance and ATTD of dietary fiber components, the lactate and propionate concentrations in ileal digesta and the butyrate, valerate and total SCFA concentrations in feces. There were interactions (p<0.05) between BW and fiber sources on the AID of GE, OM, EE, SDF, hemicellulose, the ATTD of EE, TDF, and IDF, and the hindgut disappearance of SDF and hemicellulose. Conclusion: Increasing BW mainly improved the digestibility of dietary fiber fractions, and the dietary fiber sources influenced the digestibility of almost all the dietary nutrients in growing pigs.
In this study, we measured antioxidant activity as DPPH radical scavenging and the total polyphenol content of pulverized and lyophilized oak pollens inoculated with fungi to confirm the husk removal effect. The total polyphenol content of oak pollen was highest in lyophilized pollen medium inoculated with Armillaria mellea, and was lowest in pollen inoculated with Lentinula edodes. Total polyphenol content of the lyophilized pollen was higher than that of the refined pollen and the pulverized pollen in oak pollen germinated with A. mellea. The total polyphenol content of the lyophilized oak pollen germinated with A. mellea was 1.4-fold higher than that extracted with water. Measurement of antioxidant activity using the DPPH (2, 2 diphenyl-1-picrylhydrazyl) free radical scavenging method showed that the lyophilized oak pollen germinated with A. mellea had the highest and that germinated with L. edodes was lowest in antioxidant activities. The lyophilized oak pollen germinated with A. mellea was 2 to 4 times higher than that extracted with water in the antioxidant activity of DPPH free radical scavenging. Many germinated cells were formed around pore of acorn pollen inoculated with L. edodes, while those were formed at the end of hyphae derived from oak pollen inoculated with A. mellea.
The influences of applications of organochlorine insecticide (HCH: Hexachlorocyclohexane, 10 ppm), fungicide (TPN: Tetrachloroisophthalonitrile, 40 ppm) and manure ($3Kg/m^2$) each or together on changes in soil microflora for consecutive years were investigated in the experimental field plots. The insecticide had a little effect on soil microbial numbers. In particular, the number of total bacteria, Gram-negative bacteria and fungi were gradually increased at the latter stage of the consecutive application, but the number of sporeforming bacteria reduced. The fungicide reduced the counts of sporeforming bacteria, actinomycetes and fungi respectively, whereas increased prominently the counts of total bacteria and Gram-negative bacteria. TPN-resistant bacteria, particulary TPN-resistant Gram-negative bacteria were gradually accumulated by the long-term application of TPN, and further the number of TPN-resistant total bacteria and the of TPN-resistant Gram-negative bacteria correlated fairly well during all the period. The influences of combined applications of both HCH and TPN on the number of soil microorganisms were equal to the respective sums of the effects of single application of each pesticide. The combined application of manure and these pesticides elevated the increasing extents of microbial numbers, while weakened the detrimental efforts of these pesticides on microbial numbers. These data suggest that the long-term application of these materials have resulted in the remarkable changes of composition of soil microflora.
This experiment was conducted to find out the changes in microflora of submerged soil uncultivated rice plant by application of CNP herbicide (2, 4, 6 - Trichlorophenyl-4-Nitrophenyl ether) under conditions applied with compost, rice straw, glucose or without organic material. The soil, sandy loam textured was incubated in green house for 66 days. Sampling and analysis of microorganisms were carried out during submergence periods. The results were summarilized as follows. 1. Number of aerobic total bacteria was increased by application of CNP herbicide during submerbed 50 days, afterthen, could not seen the difference. The application of rice straw increased number of aerobic bacteria regardless of CNP herbicide application or not, but glucose tended to decrease. 2. Number of Fungi was constantly maintained at $8-20{\times}10^3$ levels during period of submergence regardless of application of CNP herbicide and organic materials or not. 3. The CNP herbicide application tended to decrease the number of actinomycetes, particularly, in the treatments without organic substances and rice straw were remarkably decreased. 4. Anaerobic-and gram-negative bacteria populations were not showed any difference by application of herbicide and organic materials. 5. The ratios of aerobic bacteria to fungi and aerobic bacteria to actinomycetes appeared high values by application of herbicide and of organic substances. 6. At 66 days after submergence, the ratio of chromogenic actinomycetes to the total number of actinomycetes was lowered in application of herbicide. On the other hand, the percentage of both pretense-positive and cellulase-positive actinomycetes to the total isolates were higher in the treatment with herbicide than An without herbicide, particularly. The ratios of pretense-positive actinomycetes were high in the rice straw application regardless of herbicide application or not, but cellulase-positive actinomycetes was not remarkably difference.
Kim Dong-Ho;Song Hyun-Pa;Lee You-Seok;Cha Bo-Suk;Kim Byung-Keun;Byun Myung-Woo
Food Science and Preservation
/
v.11
no.3
/
pp.394-399
/
2004
The effects of gamma irradiation to improve the hygienic quality and microbiological shelf stability of whole baked egg were investigated by comparison with autoclaving process. The contamination levels of coliform, total aerobic bacteria and fungal group in a fresh egg were 10$^{5}$ CFU/g, 10$^{2}$ CFU/g and 10$^{1}$ CFU/g, respectively. After baking process, total aerobic bacteria and fungi were not exceeded to 10$^{1}$ CFU/g. Also, coliform was not detected under the aseptic process. However, cell counts of the baked egg after packaging reached to 10$^{4}$ CFU/g of total aerobic bacteria, 10$^{1}$ CFU/g of coliform, and 10$^{2}$ CFU/g of fungi. Therefore, it was assumed that microbial contamination of baked and packaged egg was mainly originated from an environmental uptake during packaging process. Microbiological shelf stability of the non sterilized control was about a week. Whereas, the baked eggs irradiated at more than 5 kGy were stable over 12 weeks at ambient condition as like those being autoclaved. Analytical texture profIle was stable within 10 kGy, but it became hardened in the sample treated with autoclaving. About 67$\%$ of panelists identified a sensory difference between non-irradiated and 10 kGy-irradiated sample. The baked egg irradiated at 10 kGy and autoclaved had lower acceptability than the control or samples irradiated lower than 5 kGy. Therefore, it was considered that optimal irradiation dose for radiation sterilization of baked and packaged egg was 5 kGy. At that point, it was recommended that appropriate microbiological shelf-life was 12 weeks at ambient condition.
Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.
Studies were conducted during 2 months from May of 1997 to evaluate the effects of pig manure compost(PMC) on soil microbial flora. To do so, a field experiment of Chinese cabbage(Brassica campestris L.) was conducted in a randomized block design on a sandy loam soil and microbial floral characteristics in soils were analyzed. Treatments to control included the application of PMC at (A) $8Mg\;ha^{-1}$CM-8), (B) $29Mg\;ha^{-1}$(CM-2,9), and (C) $57Mg\;ha^{-1}$(CM-57), and of chemical fertilizer(D) at $320N-80P_2O_5-200K_2O\;kg\;ha^{-1}$(NPK). In each treatment, the rhizosphere and non-rhizosphere soils were tested for the analysis of microbial populations. The populations of bacteria, actinomycetes, and fungi increased in soils with the applications of PMC and chemical fertilizer, but that of Bacillus sp. decreased. However, the population of fluorescent Pseudomonas sp. was reduced in NPK plots only. With increasing application rates of PMC, the number of colony forming units(cfu) of bacteria (Pseudomonas sp. and actinomycetes) and fungi increased. in all PMC-treated plots, the population density peaked at early growth stage for bacteria(including Bacillus sp.), at late growth for fluorscent Pseudomonas sp., and at harvest for fungi and actinomycetes. The rhizosphere effect was greatest for fluorscent Pseudomonas sp. As the application rates of PMC increased, Total N, organic matter, available phosphate, and exchangeable -K, -Ca, and -Mg increased compared to control, but soil pH was lowered. In NPK plots, EC was 3.4-fold and exchangeable K was 5-fold higher than control.
Journal of The Korean Society of Grassland and Forage Science
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v.33
no.1
/
pp.21-29
/
2013
The studies were conducted to investigate real condition of mycotoxin contamination in the fields before harvest and by the storages of rice straw used as roughage in Korea. It was found mycotoxin contamination before harvest of rice straw that the rice plants were detected deoxynivalenol at the initial growth, ochratoxin A and deoxynivalenol at the middle growth, and deoxynivalenol and zearalenone at the harvest periods in the fields. Also, the rice plants were contaminated by various fungi such as Fusarium sp., Fusarium proliferatum, Penicillium sp., Gibberella sp., Gibberella zeae, Mucor circinelloides and Aspergillus oryzae. The levels of fungal contamination were $10^{3-4}$ cfu/g at the initial growth, and $10^{4-5}$ cfu/g at the middle growth and harvest periods. All storage types of rice straw were contaminated with zearalenone, deoxynivalenol and ochratoxins A. The samples of rice straw contaminating mycotoxins were account for 3% in bundle rice straw, and 38% in both types of square rice straw and rice round bale silage, respectively. When 105 samples of rice bale silage were analyzed for mycotoxins depending on the regional area, mycotoxin contamination was found in 46% of total samples in Korea. Regional contaminations of mycotoxins were respectively 48, 33, 40, 50 and 57% of samples in Gyeonggi, Gangwon, Chungcheng, Yeongnam and Honam area. Rice round bale silage was contaminated by three kinds of mycotoxins (zearalenone, deoxynivalenol and ochratoxinsA) in the all of area without Chungcheong area where was contaminated zearalenone and deoxynivalenol. Ochratoxins A, deoxynivalenol and zearalenone were respectively determinated with the average levels of 2.6, 413 and $338{\mu}g/kg$ in rice round bale silage for the overall area, even if it was some difference depending on each regional area. Therefore, the above results clearly show that the rice straws were exposed to the contamination by mycotoxin and mycotoxigenic fungi before harvest in the fields, and mycotoxin contamination was not dependent on the regional area or the storage types such as bundle rice straw, square rice straw and rice round bale silage.
This study was conducted to investigate the effects of eucalyptus (E. Camaldulensis) crude oils (EuO) supplementation on voluntary feed intake and rumen fermentation characteristics in swamp buffaloes. Four rumen fistulated swamp buffaloes, body weight (BW) of $420{\pm}15.0$ kg, were randomly assigned according to a $2{\times}2$ factorial arrangement in a $4{\times}4$ Latin square design. The dietary treatments were untreated rice straw (RS) without EuO (T1) and with EuO (T2) supplementation, and 3% urea-treated rice straw (UTRS) without EuO (T3) and with EuO (T4) supplementation. The EuO was supplemented at 2 mL/h/d in respective treatment. Experimental animals were kept in individual pens and concentrate mixture was offered at 3 g/kg BW while roughage was fed ad libitum. Total dry matter and roughage intake, and apparent digestibilites of organic matter and neutral detergent fiber were improved (p<0.01) by UTRS. There was no effect of EuO supplementation on feed intake and nutrient digestibility. Ruminal pH and temperature were not (p>0.05) affected by either roughage sources or EuO supplementation. However, buffaloes fed UTRS had higher ruminal ammonia nitrogen and blood urea nitrogen as compared with RS. Total volatile fatty acid and butyrate proportion were similar among treatments, whereas acetate was decreased and propionate molar proportion was increased by EuO supplementation. Feeding UTRS resulted in lower acetate and higher propionate concentration compared to RS. Moreover, supplementation of EuO reduced methane production especially in UTRS treatment. Protozoa populations were reduced by EuO supplementation while fungi zoospores remained the same. Total, amylolytic and cellulolytic bacterial populations were increased (p<0.01) by UTRS; However, EuO supplementation did not affect viable bacteria. Nitrogen intake and in feces were found higher in buffaloes fed UTRS. A positive nitrogen balance (absorption and retention) was in buffaloes fed UTRS. Supplementation of EuO did not affect nitrogen utilization. Both allantoin excretion and absorption and microbial nitrogen supply were increased by UTRS whereas efficiency of microbial protein synthesis was similar in all treatments. Findings of present study suggested that EuO could be used as a feed additive to modify the rumen fermentation in reducing methane production both in RS and UTRS. Feeding UTRS could improve feed intake and efficiency of rumen fermentation in swamp buffaloes. However, more research is warranted to determine the effect of EuO supplementation in production animals.
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