This study was conducted to screen dual-purpose lactic acid bacteria (LAB) from uncontrolled farm-scale silage, and then we confirmed their effects on corn silage. The LAB were isolated from eight farm-scale corn silages, and then we screened the antifungal activity against Fusarium graminearum and the carboxylesterase activity using spectrophotometer with p-nitrophenyl octanoate as substrate and McIlvane solution as buffer. From a total of 25 isolates, 5M2 and 6M1 isolates were selected as silage inoculants because presented both activities of antifungal and carboxylesterase. According 16S rRNA gene sequencing method, 5M2 isolate had 100.0% similarity with Lactobacillus brevis, and 6M1 isolate had 99.7% similarity with L. buchneri. Corn forage was ensiled in bale silo (500 kg) for 72 d without inoculant (CON) or with mixture of selected isolates at 1:1 ratio (INO). The INO silage had higher nutrient digestibility in the rumen than CON silage. Acetate was higher and yeasts were lower in INO silage than in CON silage on the day of silo opening. In all days of aerobic exposure, yeasts were lower in INO silage than CON silage. The present study concluded that Lactobacillus brevis 5M2 and L. buchneri 6M1 confirmed antifungal and carboxylesterase activities on farm-scale corn silage.
1 June 2020 in Asian-Australasian Journal of Animal Sciences
Objective: This study was conducted to confirm the effects of new inoculants producing-antifungal or esterase substances on rye silage and its rumen fermentation indices by comparing wild with mutated types.Methods: Rye harvested at dough stage was ensiled into 3 L mini bucket silo (1 kg) for 90 d in triplicate following: distilled water at 20 μL/g (CON); Lactobacillus brevis 100D8 (AT) and its inactivation of antifungal genes (AT-m) at 1.2×105 cfu/g, respectively; and Leuconostoc holzapfelii 5H4 (FD) and its inactivation of esterase genes (FD-est) at 1.0×105 cfu/g, respectively. After silo opened, silage was sub-sampled for the analysis of ensiling quality and its rumen fermentation indices.Results: Among the wild type inoculants (CON vs AT vs FD), FD inoculant had higher (p<0.05) in vitro digestibilities of dry matter and neutral detergent fiber, the total degradable fraction, and total volatile fatty acid in rumen, while AT inoculant had higher (p<0.05) lactate, acetate, and lactic acid bacteria in silage. Silage pH and the potentially degradable fraction in rumen increased (p<0.05) by inactivation of antifungal activity (AT vs AT-m), but lactate, acetate, and lactic acid bacteria of silage decreased (p<0.05). In silage, acetate increased (p<0.05) by inactivation of esterase activity (FD vs FD-est) with decreases (p<0.05) of pH, ammonia-N, lactate, and yeast. Moreover, inactivation of esterase activity clearly decreased (p<0.05) in vitro digestibilities of dry matter and neutral detergent fiber, the total degradable fraction, and total volatile fatty acid in the rumen.Conclusion: This study concluded that FD inoculant confirmed esterase activity on rye silage harvested at dough stage, while AT inoculant could not be confirmed with antifungal activity due to the absence of mold in all silages.
1 January 2019 in Journal of Applied Animal Research
This study estimated the effects of a new inoculant producing antifungal and esterase activity on quality of high moisture sorghum-sudangrass (SS) silage with two different hybrids (SX-17 and Speed-up). The SS hybrids were chopped, treated without an inoculant (CON) and with an inoculant containing Lactobacillus plantarum R48-27 and Lactobacillus buchineri R4-26 at ratio 1:1 (INO), then ensiled into 20-L mini silo in quadruplicate for 60 days. After ensiling, silage was placed under aerobic condition for 8 days to estimate yeast and mold. The INO silages had higher (P < 0.05) dry matter, crude protein, neutral detergent fibre, and acid detergent fibre than those of CON silages. The INO silages also had higher (P < 0.05) pH and acetate, but lower (P < 0.05) ammonia-N, butyrate, and lactate to acetate ratio than those of CON silages. Applied INO in both hybrids had lower (P < 0.05) yeast after 4–8 days of aerobic exposure than CON. In rumen, INO silages had higher (P < 0.05) in vitro dry matter digestibility, pH, ammonia-N, and acetate than those of CON silages. In conclusion, the new inoculant application improved not only fermentation quality, but also rumen digestibility of high moisture SS silage.
The present study aimed to investigate effects of dual-purpose inoculants (antifungal and carboxylesterase activities) not only on corn silage quality, but also its shelf life against mold contamination at feed-out phase. Corn forage was ensiled for 252 d with different inoculants of the following: control (CON), Lactobacillus brevis 5M2 (5M), Lactobacillus buchneri 6M1 (6M), and mixture of 5M and 6M at 1:1 ratio (MIX). After ensiling, corn silage was contaminated with Fusarium graminearum. Silages applied inoculants had positive effects by increased organic acid and lactic acid bacteria, and decreased undesirable microbes. At feed-out phase, contamination of F. graminearum into corn silage had a negative effect on aerobic stability caused by increased growth of undesirable microbes. However, silages applied inoculants had positive effects by decreased undesirable microbes and extended lactic acid bacteria and aerobic stability. Generally, MIX silage presented better effects on organic acid production, rumen degradation, inhibition of undesirable microbes, and aerobic stability than 5M silage and 6M silage. The present study concluded that application of inoculants into corn silage had positive effects on fermentation characteristics and extended shelf life against mold contamination at feed-out phase. A mixed inoculant appeared to have better effects of antifungal and carboxylesterase than a single inoculant.
This study investigated the effects of essential oil (EO) from three Korean wormwood (Artemisia Montana) plants on in vitro ruminal digestibility, fermentation, and microbial diversity. Dried (0.5 g) soybean meal (SBM) or bermudagrass hay (BGH) were incubated in buffered rumen fluid (40 mL) for 72 h with or without EO (5 mg/kg) from Ganghwa (GA), Injin (IN), or San (SA) wormwood (Experiment 1). Both SA and IN improved (p < 0.05) dry matter digestibility (DMD) of BGH, while GA reduced (p < 0.05) total short-chain fatty acid of BGH and SBM. Besides, SA increased (p < 0.05) numbers of Ruminococcus albus and Streptococcus bovis in SBM. Experiment 2 examined different doses (0, 0.1, 1, and 10 mg/kg) of SA, the most promising EO from Experiment 1. Applying SA at 10 mg/kg gave the highest DMD (L; p < 0.01) and neutral detergent fiber (Q; p < 0.05) digestibility for BGH. Applying SA at 1 mg/kg gave the highest R. albus population (Q; p < 0.05) in SBM. Therefore, SA was better than GA and IN at improving rumen fermentation, and the 0.1 to 1 and 10 mg/kg doses improved ruminal fermentation and in vitro digestibility of SBM and BGH, respectively.
1 December 2018 in Asian-Australasian Journal of Animal Sciences
This study was conducted to evaluate the effect of homo or hetero fermentative inoculants on fermentation quality and aerobic stability of sweet potato vine (SPV) silage containing Italian ryegrass hay as moisture absorbent. The SPV was harvested at 15% dry matter, mixed with Italian ryegrass hay at 1:1 ratio on a fresh weight basis, and chopped to 3 to 5 cm length. After then, the chopped forage mixture was ensiled into 20-L mini silos in quadruplicate for 7, 48, and 100 days after application of microbial inoculants at 1.2×105 colony forming units (cfu)/g of forage following: no inoculant (CON), Lactobacillus plantarum as a homo fermentative (LP), Lactobacillus buchneri as a hetero fermentative (LB), and mixture of LP and LB at 1:1 ratio as a combo fermentative (MIX). The LP and MIX silages had lowest pH (p<0.001) on 7 and 48 days, while MIX and CON silages had greatest lactate concentrations (p<0.05) on 7 and 48 days, respectively. Acetate concentrations were highest (p<0.01) in LB and MIX silages on 7 days, and in LB silage on 48 days, while lactate to acetate ratios were lowest (p<0.001) in LB silages. The chemical compositions and nutrient digestibility of silage ensiled for 100 days was not affected by inoculants. On 100 days of ensiling, LB silage had lowest (p<0.01) lactate concentration and lactate to acetate ratio, but highest acetate concentration. Aerobic stability was highest (p<0.001) in LB silage followed in MIX silage. On contrast, LB silage had lowest (p<0.05) lactic acid bacteria and mold. The results indicated that application of LB solely had a better effect on aerobic stability than not only LP, but also MIX. However, LP application did not show beneficial effects from the viewpoints of fermentation quality and aerobic stability compared to CON.