• Asian-Australasian Journal of Animal Sciences
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• v.13 no.1
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• pp.23-30
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• 2000

The effects of grass lipids and long chain fatty acids (LCFA; palmitic, stearic and oleic acids), at low concentrations (0.001~0.02%), on the growth and enzyme activity of two strains of anaerobic fungi, monocentric strain Piromyces rhizinflata B157 and polycentric strain Orpinomyces joyonii SG4, were investigated. The addition of grass lipids to the medium significantly (p<0.05) decreased filter paper (FP) cellulose digestion, cellulase activity and fungal growth compared to control treatment. However, LCFA did not have any significant inhibitory effects on fungal growth and enzyme activity, which, however, were significantly (p<0.05) stimulated by the addition of oleic acid as have been observed in rumen bacteria and protozoa. This is the first report to our knowledge on the effects of LCFA on the rumen anaerobic fungi. Continued work is needed to identify the mode of action of LCFA in different fungal strains and to verify whether these microorganisms have ability to hydrogenate unsaturated fatty acids to saturated fatty acids.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.10
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• pp.1416-1423
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• 2013

The metabolomic profile of the anaerobic fungus Piromyces sp. F1, isolated from the rumen of goats, and how this is affected by the presence of naturally associated methanogens, was analyzed by nuclear magnetic resonance spectroscopy. The major metabolites in the fungal monoculture were formate, lactate, ethanol, acetate, succinate, sugars/amino acids and ${\alpha}$-ketoglutarate, whereas the co-cultures of anaerobic fungi and associated methanogens produced citrate. This is the first report of citrate as a major metabolite of anaerobic fungi. Univariate analysis showed that the mean values of formate, lactate, ethanol, citrate, succinate and acetate in co-cultures were significantly higher than those in the fungal monoculture, while the mean values of glucose and ${\alpha}$-ketoglutarate were significantly reduced in co-cultures. Unsupervised principal components analysis revealed separation of metabolite profiles of the fungal mono-culture and co-cultures. In conclusion, the novel finding of citrate as one of the major metabolites of anaerobic fungi associated with methanogens may suggest a new yet to be identified pathway exists in co-culture. Anaerobic fungal metabolism was shifted by associated methanogens, indicating that anaerobic fungi are important providers of substrates for methanogens in the rumen and thus play a key role in ruminal methanogenesis.

• Journal of The Korean Society of Grassland and Forage Science
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• v.39 no.3
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• pp.132-140
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• 2019

Fermented total mixed ration (TMR) is a novel feed for ruminants in South Korea. The purpose of this study was to evaluate the effects of lactic acid bacteria (LAB) on the quality of TMR and in vitro ruminal fermentation. Strains of three LAB spp. (Lactobacillus plantarum, L. brevis, L. mucosae) were used in fermentation of TMR. Inoculations with the three LAB spp. lowered pH and increased concentrations of lactic acid, acetic acid, and total organic acid compared to non-LAB inoculated control (only addition of an equivalent amount of water) (p<0.05). Bacterial composition indicated that aerobic bacteria and LAB were higher. However, E. coli were lower in the fermented TMR than those in the control treatment (p<0.05). Among the treatments, L. brevis treatment had the highest concentration of total organic acid without fungus detection. Gas production, pH, and ammonia-nitrogen during ruminal in vitro incubation did not differ throughout incubation. However, ruminal total VFA concentration was higher (p<0.05) in the LAB spp. treatments than the control treatment at 48 hours. Overall, the use of L. brevis as an inoculant for fermentation of high moisture. TMR could inhibit fungi growth and promote lactic fermentation, and enhance digestion in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.3
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• pp.357-365
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• 2019

Objective: The experiment was conducted to evaluate the effects of four fungal pretreatments on the nutritional value of Camellia seed residues, and to evaluate the feeding value of pretreated Camellia seed residues for ruminants. Methods: Camellia seed residues were firstly fermented by four lignin degrading fungi, namely, Phanerochaete chrysosporium (P. chrysosporium)-30942, Trichoderma koningiopsis (T. koningiopsis)-2660, Trichoderma aspellum (T. aspellum)-2527, or T. aspellum-2627, under solid-state fermentation (SSF) conditions at six different incubation times. The nutritional value of each fermented Camellia seed residues was then analyzed. The fermentation profiles, organic matter degradability and metabolizable energy of each pre-treated Camellia seed residue were further evaluated using an in vitro rumen fermentation system. Results: After 5 days of fermentation, P. chrysosporium-30942 had higher degradation of lignin (20.51%), consumed less hemicellulose (4.02%), and the SSF efficiency reached 83.43%. T. koningiopsis-2660 degraded more lignin (21.54%) and consumed less cellulose (20.94%) and hemicellulose (2.51%), the SSF efficiency reached 127.93%. The maximum SSF efficiency was 58.18% for T. aspellum-2527 and 47.61% for T. aspellum-2627, appeared at 30 and 15 days respectively. All the fungal pretreatments significantly improved the crude protein content (p<0.05). The Camellia seed residues pretreated for 5 days were found to possess significantly increased organic matter degradability, volatile fatty acid production and metabolizable energy (p<0.05) after the treatment of either P. chrysosporium-30942, T. koningiopsis-2660 or T. aspellum-2527. The fungal pretreatments did not significantly change the rumen fermentation pattern of Camellia seed residues, with an unchanged ratio of acetate to propionate. Conclusion: The fungi showed excellent potential for the solid-state bioconversion of Camellia seed residues into digestible ruminant energy feed, and their shorter lignin degradation characteristics could reduce loss of the other available carbohydrates during SSF.

• Journal of Animal Science and Technology
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• v.45 no.6
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• pp.1019-1030
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• 2003

The study was conducted to isolate and identify highly fibrolytic anaerobic fungi from the guts of a Hanwoo steer and a Korean native goat, and then investigate the characterization of cellulolytic activity of an anaerobic fungus. Twenty-one anaerobic fungal colonies were isolated in the study, in which 16 colonies were isolated from the rumen contents of the Hanwoo steer and 5 colonies from the duodenal fluids of the Korean native goat. Four anaerobic fungi were selected based on higher cellulolytic enzyme activities to identify under a optical microscope. NLRI-M003 and -T004 belong to Neocallimastix genus and NLRI-M014 belongs to Piromyces genus based on the morphology of their thallus, sporangia, rhizoid and the number of flagella. NLRI-M001 appeared to be an unknown strain of anaerobic fungi due to its different morphology from existing types of anaerobic fungi, though the morpholgoy is similar to Orpinomyces sp. Supplementation of 2% anaerobic fungal culture(NLRI-M003) in rumen-mixed microorganisms increased in vitro DM degradability of rice straw and filter paper up to 4 and 11%, respectively, compared with non-supplementation(control). CMCase and xylanase activities in in vitro culture were also higher in 2% fungal supplementation than controls in both rice straw and filter paper substrates.

• Microbiology and Biotechnology Letters
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• v.25 no.4
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• pp.346-353
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• 1997

During the screening of cellulase producing microorganisms, a fungal strain C-4 was selected from etiolated leaves. Based on taxonomic studies, the fungus could be classified as a strain of Trichoderma sp. When the strain C-4 was cultured in Mandels' media at 28$circ$C for 6 days, the enzyme activities detected in broth were as follows: 8.2 U/ml (28.1 U/mg) of CMCase activity, 0.75 U/ml (2.58 U/mg) of Avicelase activity, 1.67 U/ ml (5.68 U/mg) of $eta$-glucosidase activity. The optimum pH for enzyme induction was 6.2. The crude enzyme retained 100% of its original CMCase activity at 50$circ$C for 1 hr (pH 5.0), and at 4$circ$C for 24 hrs (pH 5.0). There was no effect on the CMCase activity in the presence of 1 mM of CsCl, LiCl, MgCl$_{2}$, and FeCl$_{2}$, respectively. When the crude enzyme was treated with trypsin and chymotrypsin (2% W/w) for 10 minutes, the remaining CMCase activity was 70%, but there was no further loss of activity for 60 minutes treatment at 30$circ$C. The crude enzyme showed the synergism with rumen fluid for the hydrolysis of Avicel and CMC by 118% and 130%, respectively.

• Journal of Animal Science and Technology
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• v.48 no.3
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• pp.415-424
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• 2006

The purpose of this study was to investigate the fungal growth and enzyme production under different carbohydrate substrate conditions. The anaerobic fungus Neocallimastix sp. NLRI-3 isolated from the rumen of Korean native goat was incubated with different carbohydrate media containing 0.2% of glucose, starch, rice straw, filter paper, carboxymethyl cellulose(CMC), Sigmacell cellulose, xylan or xylose, respectively. The culture head gas production was the highest in the culture of filter paper medium, and the lowest in the culture of CMC medium at 96h incubation (P<0.05). The fungal zoospore production reached peak at 72h incubation, and its number was the highest in rice straw medium among the treatments (P<0.05). At 96h incubation, carboxymethyl cellulase(CMCase) activity was the highest in the culture of filter paper medium and the lowest in the culture of starch medium (P<0.05). While xylanase activity was the highest in the culture of rice straw medium and the lowest in the culture of xylose medium(P<0.05) at 72h incubation. There were no differences in culture supernatant protein expression among the treatments. However, the patterns of enzyme expression were different among the treatments with zymogram analysis. Six CMCases and 4 xylanase were detected from the results of zymogram analysis. Therefore the present study indicating that the fungal enzyme expression could be stimulated with insoluble substrates in the culture medium.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.4
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• pp.500-508
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• 2016

This study was conducted to investigate the effects of Momordica charantia saponin (MCS) on ruminal fermentation of maize stover and abundance of selected microbial populations in vitro. Five levels of MCS supplements (0, 0.01, 0.06, 0.30, 0.60 mg/mL) were tested. The pH, $NH_3-N$, and volatile fatty acid were measured at 6, 24, 48 h of in vitro mixed incubation fluids, whilst the selected microbial populations were determined at 6 and 24 h. The high dose of MCS increased the initial fractional rate of degradation at t-value = 0 ($FRD_0$) and the fractional rate of gas production (k), but decreased the theoretical maximum of gas production ($V_F$) and the half-life ($t_{0.5}$) compared with the control. The $NH_3-N$ concentration reached the lowest concentration with 0.01 mg MCS/mL at 6 h. The MSC inclusion increased (p<0.001) the molar proportion of butyrate, isovalerate at 24 h and 48 h, and the molar proportion of acetate at 24 h, but then decreased (p<0.05) them at 48 h. The molar proportion of valerate was increased (p<0.05) at 24 h. The acetate to propionate ratio (A/P; linear, p<0.01) was increased at 24 h, but reached the least value at the level of 0.30 mg/mL MCS. The MCS inclusion decreased (p<0.05) the molar proportion of propionate at 24 h and then increased it at 48 h. The concentration of total volatile fatty acid was decreased (p<0.001) at 24 h, but reached the greatest concentration at the level of 0.01 mg/mL and the least concentration at the level of 0.60 mg/mL. The relative abundance of Ruminococcus albus was increased at 6 h and 24 h, and the relative abundance of Fibrobacter succinogenes was the lowest (p<0.05) at 0.60 mg/mL at 6 h and 24 h. The relative abundance of Butyrivibrio fibrisolvens and fungus reached the greatest value (p<0.05) at low doses of MCS inclusion and the least value (p<0.05) at 0.60 mg/mL at 24 h. The present results demonstrates that a high level of MCS quickly inhibits in vitro fermentation of maize stover, while MCS at low doses has the ability to modulate the ruminal fermentation pattern by regulating the number of functional rumen microbes including cellulolytic bacteria and fungi populations, and may have potential as a feed additive applied in the diets of ruminants.