The effect of anaerobic fungal inoculation on the fermentation characteristics of rice straw silages.

AIMS: To identify whether the supplement of anaerobic fungi isolates with cellulolytic activities accelerates the silage fermentation. METHODS AND RESULTS: Three fungal isolates with the highest cellulolytic activities among 45 strains of anaerobic fungal stock in our laboratory were selected and used as silage inoculants. The rice straw (RS) was ensiled for 10, 30, 60, 90 and 120 days with four treatments of anaerobic fungi derived from the control (no fungus), Piromyces M014 (isolated from the rumen of the Korean native goat), Orpinomyces R001 (isolated from the duodenum of Korean native cattle) and Neocallimastix M010 (isolated from the guts of termites), respectively. The silages inoculated with pure strains of fungi showed a higher fungal population (P < 0.05) when compared to the control silage. In situ ruminal DM disappearance of RS silage (RSS) was improved with fungal treatment. SEM observation showed live fungal cells inoculated in RS could survive during the ensiling process. Overall, this study indicated that the inoculation of anaerobic fungi decreased the cell wall content of the RSS and increased in situ dry matter disappearance. CONCLUSIONS: The supplementation of anaerobic fungi isolates to RSS as a silage inoculant improves the RSS quality. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study showing the potential application of supplement of anaerobic fungi isolated from the guts may be applied industrially as an alternate feed additive that improves the silage quality.

Fibrolytic potential of anaerobic fungi (Piromyces sp.) isolated from wild cattle and blue bulls in pure culture and effect of their addition on in vitro fermentation of wheat straw and methane emission by rumen fluid of buffaloes.

BACKGROUND: Ten isolates of anaerobic fungi of Piromyces genus from wild cattle and blue bulls (five isolates from each host species) were evaluated for their fibrolytic ability in pure culture, their suitability for use as a microbial additive in buffaloes and their effect on methane emission. RESULTS: In pure culture, only two out of five isolates from wild cattle degraded wheat straw efficiently, whereas all five isolates from wild blue bulls did. Isolate CF1 (from cattle) showed the highest apparent digestibility (53.4%), true digestibility (70.8%) and neutral detergent fibre digestibility (75.0%) of wheat straw after 5 days of incubation. When added to buffalo rumen fluid, all five isolates from cattle increased (P < 0.05) in vitro apparent digestibility of wheat straw compared with the control (received autoclaved culture), but all five isolates from blue bulls failed to influence in vitro digestibility of wheat straw. Isolate CF1 showed the highest stimulating effect on straw digestion by buffalo rumen fluid microbes and increased apparent digestibility (51.9 vs 29.4%, P < 0.05), true digestibility (57.9 vs 36.5%, P < 0.05) and neutral detergent fibre digestibility (51.5 vs 26.9%, P < 0.05) of wheat straw compared with the control after 24 h of fermentation. There were also significant increases in fungal count and enzyme activities of carboxymethylcellulase and xylanase in the CF1-added group compared with the control group. Gas and methane production g(-1) truly digested dry matter of straw were comparable among all groups including the control. CONCLUSION: Wild cattle and blue bulls harbour some anaerobic fungal strains with strong capability to hydrolyse fibre. The fungal isolate CF1 has high potential for use as a microbial feed additive in buffaloes to improve digestibility of fibrous feeds without increasing methane emission per unit of digested feed.

Effect of anaerobic fungi on in vitro feed digestion by mixed rumen microflora of buffalo.

Five strains of anaerobic fungi isolated from the faeces of wild (hog deer, Cervus porcinus; blackbuck, Antelope cervicapra; spotted deer, Axis axis; nilgai, Baselophus tragocamelus) and rumen liquor of domestic (sheep, Ovies aries) ruminants showing high fibrolytic enzyme producing ability were added to mixed rumen microflora of buffalo to study their effect on the digestibility of lignocellulosic feed (wheat straw and wheat bran in the ratio of 80:20), enzyme production and fermentation end products in in vitro conditions. Among the 5 isolates studied, FNG5 (isolated from nilgai) showed the highest stimulating effect on apparent digestibility (35.31 +/- 1.61% vs. 28.61 +/- 1.55%; P < 0.05), true digestibility (43.64 +/- 1.73% vs. 35.37 +/- 1.65%; P < 0.01), neutral detergent fiber digestibility (29.30 +/- 2.58% vs. 18.47 +/- 2.12; P < 0.01) of feed 24 h after inoculation compared to the control group. The production of carboxymethyl cellulase, xylanase, acetyl esterase and beta-glucosidase was significantly (P < 0.05) higher in the FNG5 inoculated incubation medium. There was no improvement in the digestibility and enzyme production on the addition of the other 4 isolates. Total volatile fatty acid levels as well as the concentration of acetate, propionate, isobutyrate and valerate were significantly higher in the FNG5 added group as compared to the control group. The fungal isolate FNG5 from nilgai, a wild ruminant, was found to be superior to the other isolates tested and appears to have a potential to be used as a feed additive for improving fiber degradation in domestic ruminants.

Effect of phenolic monomers on biomass and hydrolytic enzyme activities of an anaerobic fungus isolated from wild nil gai (Baselophus tragocamelus).

AIMS: To test the anaerobic fungus, Piromyces sp. FNG5, for its tolerance to phenolic monomers released in the rumen by degradation of lignocellulosic poor-quality feeds. METHODS AND RESULTS: Effects of phenolic monomers on biomass and fibrolytic enzyme activities of a pure culture of lignocellulolytic anaerobic fungus (Piromyces sp. FNG5) isolated from faeces of wild nil gai (blue bull, Baselophus tragocamelus) were evaluated. There was a reduction in fungal biomass at 1 mm concentration of catechol with complete inhibition at 10 mm. p-Coumaric acid caused a reduction in biomass at 10 mm and no growth was observed above 20 mm concentration. The fungal isolate could tolerate up to 5 mm of ferulic acid without any reduction in biomass level, and was able to grow to some extent up to the highest level of ferulic acid tested (20 mm). Vanillic acid had no effect on biomass of the fungus even up to 50 mm level. The phenolic monomers varied in their potential to inhibit the secretion of carboxymethyl cellulase, xylanase, beta-glucosidase and acetyl esterase activities with catechol being the most inhibitory and vanillic acid being the least inhibitory. After 14 days of incubation, 38.49-65.14%p-Coumaric acid, 65.22-74.10% ferulic acid and 34.13-66.78% vanillic acid disappeared from the medium under anaerobic conditions. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: It is concluded that the anaerobic fungus Piromyces sp. FNG5 is tolerant to phenolic monomers and has ability to degrade them. Therefore, such anaerobic fungi may play an important role in fibre degradation in the rumen.