The effects of new strains of lactic acid bacteria on alfalfa silage fermentation were evaluated. The experiment was performed using a completely randomized design (with three replicates) based on a 6 × 6 factorial assay with 6 inoculants (I): Control (CTRL), Commercial inoculant (CI),Lactobacillus pentosus14.7SE (LPE),Lactobacillus plantarum3.7E (LP),Pediococcus pentosaceus14.15SE (PP), andLactobacillus plantarum3.7E + Pediococcus pentosaceus14.15SE (LP + PP), and six fermentation periods (P): 1, 3, 7, 14, 28 and 56 days. Alfalfa was wilted for 6 h in the field, which increased the dry matter content to 368 g/kg as fed. The CP and yeast population decreased during the fermentation process. Silage inoculated with the PP strain had the lowest pH values beginning at 14 d of fermentation and the lowest acetic acid concentration on the last day of fermentation. New strains more efficiently regulated enterobacteria and mold populations at days 56 and 28, respectively. Silages inoculated with the PP strain had a higher coefficient ofin vitrodry matter digestibility than LP silages. All of the tested novel strains resulted in positive effects on at least one chemical property of the silage during the fermentation process. However, the adding ofP.pentosaceuscan be indicated as the better for silage quality considering the tested treatments in the present study.

The first part of the study aimed to isolate, characterize, and identify wild lactic acid bacteria (LAB) strains from alfalfa silage produced in a tropical area. LAB strains were isolated from alfalfa silage ensiled for 1, 3, 7, 14, 28, and 56 days (d) and were identified by sequencing the 16S rRNA gene. The second part aimed to investigate the effects of wild LAB strains on the nutritive and fermentative characteristics of alfalfa silage. This trial was conducted according to a completely randomized design in a 4 × 2 factorial scheme [four inoculants (I) × two harvests (H)], (n = 4). The inoculants were: (1) no inoculant (CTRL), (2) Lactobacillus pentosus (AV 14.17); (3) L. pentosus + Lactobacillus brevis + Pediococcus acidilactici (Combo); and (4) commercial inoculant (CI). Alfalfa forage (7 kg) was ensiled in 10 L buckets and opened after 90 d. Seventy-seven strains were isolated. Pediococcus, Lactobacillus, and Weissella represented 52.0, 24.7, and 20.8% of the isolates, respectively. For the first harvest, Combo, CI, and all inoculated silages showed lower acid detergent fiber ADF, neutral detergent fiber (NDF), and ammonia nitrogen (NH₃-N), respectively. Silage fermented with AV14.17 presented greater residual water-soluble carbohydrate (WSC) in the second harvest and showed the lowest pH in both harvests. AV14.17 strain has potential as an inoculant for alfalfa silage production.

Two separate experiments were carried out to evaluate the effects of incremental doses of 10 exogenous endo-acting α-amylase and exo-acting glucoamylase; 1LAT (bacterial α-amylase), 2AK, 3AC, 4Cs4, 5Trga, 6Afuga, 7Fvga, and 10Tg (fungal α-amylases, glucoamylases, and α-glucosidase), 8Star and 9Syn (fungal amylase-mixtures; experiment 1) and three exogenous proteases; 11P14L, 12P7L, and 13P30L (bacterial proteases; experiment 2) on in vitro dry matter digestibility (IVDMD) and in vitro starch digestibility (IVSD) of mature dent corn grain using a batch culture system. Incremental doses of the exogenous enzymes (0, 0.25, 0.50, 0.75, and 1.00 mg/g of dried substrate) were applied directly to the substrate (0.5 g of ground corn, 4 mm) in sextuplicate (experiment 1) or quadruplicate (experiment 2) within F57 filter bags, which were incubated at 39 °C in buffered rumen fluid for 7 h. Rumen fluid was collected 2–3 h after the morning feeding from three lactating dairy cows and pooled. Cows were consuming a midlactation total mixed ration (TMR; 1.60 Mcal/kg DM and 15.4%; net energy of lactation and crude protein, respectively). Three independent runs were carried out for each experiment. Data were analyzed as a randomized complete block design using run as the blocking factor. Dose was used as a fixed factor while run was considered a random factor. Linear, quadratic, and cubic orthogonal contrasts were also tested. In experiment 1, enzymes 2AK, 3AC, and 10Tg did not increase (P > 0.10) IVDMD and IVSD, whereas 0.25 mg of enzymes 1LAT, 5Trga, and 8Star increased (P < 0.01) IVDMD by 23%, 47%, and 62% and IVSD by 35%, 41%, and 58%, respectively, compared with the control. Enzymes 4Cs4, 6Afuga, 7Fvga, and 9Syn linearly increased IVDMD and IVSD (P < 0.01). Greatest increases in IVDMD (82.9%) and IVSD (85.9%) resulted with 1 mg of 6Afuga compared to control. In experiment 2, the lowest dose of exogenous proteases 11P14L and 12P7L increased (P < 0.01) IVDMD by 98% and 87% and IVSD by 57% and 64%, respectively, whereas the highest dose of 13P30L increased (P = 0.02) IVDMD by 44.8% and IVSD by 30%, relative to the control. In conclusion, IVSD and IVDMD were increased by one α-amylase, certain glucoamylases, and all proteases tested, with the glucoamylase 6Afuga in experiment 1 and the neutral protease 12P7L in experiment 2, increasing IVDMD and IVSD to the greater extents. Future in vivo studies are required to validate these findings before these enzyme additives can be recommended for improving the digestibility of mature dent corn grain.

The aerobic deterioration of silage nutrients is inevitable in tropical countries, causing negative consequences in animal production systems. Aiming to minimize the losses, the effects of Lactobacillus buchneri strains on fermentation profile and aerobic stability of corn silages were evaluated. The experiment was conducted under a completely randomized design with 13 treatments and three replicates. The treatments were noninoculated, commercial L. buchneri (CI), and 11 wild strains of L. buchneri: LB-56.1, LB-56.2, LB-56.4, LB-56.7, LB-56.8, LB-56.9, LB-56.21, LB-56.22, LB-56.25, LB-56.26, and LB-56.27. The treatments could be divided into three different groups according to silage pH and acetic acid concentration. Silages inoculated with LB-56.1, LB-56.4, and LB-56.9 presented higher pH, whereas intermediate values were observed for LB-56.2, LB-56.7, and LB-56.8. The highest acetic acid production was observed for LB-56.1 and LB-56.7. On the other hand, lowest concentrations were found for CI, LB-56.22, LB-56.25, LB-56.26, and LB-56.27. Higher amounts of NH₃–N were observed for LB-56.8, LB-56.21, LB-56.22, and LB-56.27 silages than others. Silage inoculation with CI, LB-56.1, LB-56.2, LB-56.4, LB-56.8, LB-56.9, and LB-56.25 strains had higher aerobic stability than others (59.7 vs. 41.2 h). The L. buchneri strains LB-56.1, LB-56.2, LB-56.4, LB-56.8, LB-56.9, and LB-56.25 provided potential features to improve the aerobic stability of corn silage.