Seleksi Galur Lactobacillus Plantarum Untuk Inokulum Pada Pembuatan Silase

Eleven strains of Lactobacillus plantarum have been evaluated for their growth rate and ability to reduce pH during corn ensiling. Ensiling was carried out in small scale using plastic bag for 14 days. Soluble carbohydrate consumption, organic acids production and pH reduction were measured during fermentation. The growth rate of each strain was also determined in liquid medium. All of the 11 strains of L. plantarum tested were able to improve the ensiling process. However, strains T-16, T-25 and S-98 grew in liquid medium faster than the others, and were able to reduce pH of silage quicker than the other strains. The pH of silage inoculated with strain T-16, T-25 and S-98 decreased from about 5.60 to 4.23, 4.45 and 4.40, respectively, in two days, and to 3.98, 4.05 and 4.03, respectively, after 14 days fermentation. The acetic acid content of silages inoculated with those strains was low. Strains T-16 and T-25 were suitable as potential silage inoculants and their characteristics were comparable to the one isolated from a commercial silage inoculant

Supplementation of two novel probiotics in the diet of lactating dairy cows

Forty-eight multiparous Holstein cows (121 ñ 22 DIM) were used in a randomized complete block design to evaluate supplementation of two probiotic strains Pediococcus acidilactici 19839 (PED) or Bacillus subtilis 15541 (BAC) on the effect of milk yield and composition, total tract nutrient digestibility, rumen pH and volatile fatty acid concentration. Cows were housed in a free-stall barn, milked three times a day, and fed twice daily for 105 d. All treatments consisted of a basal TMR diet, top-dressed with a specific supplement: 1) control (CON) with no probiotics; 2) PED fed at 1 à1010 CFU/d; 3) BAC fed at 1 à1010 CFU/d and; 4) basal TMR supplemented with a combination of Enterococcus faecium at 1 à1010 CFU/d and yeast (PRO). Individual feed intake and milk yield data were recorded daily and averaged weekly. Two blocks contained rumen fistulated cows and were used for rumen pH measurements and rumen fluid collection on d 105. Data were analyzed using a mixed model with week, treatment and their interaction as fixed effects with pre-experiment milk yield as a covariate and cow and block as random effects. Dry matter intake was similar across treatments with an average of 24.3 ñ 0.8 kg/d. Milk yield averaged 37.4 ñ 1.4 kg/d across treatments; analyses of data in 5-wk periods showed that PED resulted in additional 3.9 ñ 2.9 kg/d for the first period compared to all other treatments. Concentration of milk fat and protein were similar across treatments with averages of 3.63 ñ 0.02 % and 3.05 ñ 0.06 %. Digestibility of dry matter, organic matter, and protein were similar across treatments and averaged 66.65 ñ 1.48 %, 68.88 ñ 1.43 %, and 67.11 ñ 1.81 %. Similarly, acid and neutral detergent fiber digestibility was similar for all treatments. Mean daily rumen pH was 5.69 ñ 0.05 across treatments. Proportions of acetate, propionate and butyrate averaged 57.1 ñ 1.8%, 26.6 ñ 2.3% and 11.1 ñ 0.7%, across treatments respectively. Although the mechanism for transient increase in milk yield remains to be elucidated, the results demonstrate that, in dairy cattle, supplementation with the specific strain P. acidilactici 19839 has the potential to improve lactation performance without detrimental effects on digestibility, rumen pH, and VFA concentration

Screening of Lactic Acid Bacteria From Rumen Liquor and King Grass Silage as Well as Their Antibacterial Activities

Probiotic is a live microbial culture which has positive effect on animal by improving the natural balance of microflora in the digestive tract. This experiment aimed to screen and identify indigenous lactic acid bacteria (LAB) from rumen liquor and king grass (Pennisetum hybrid) silage as a probiotic candidate and to evaluate their resistance in low pH, and inhibitory activities against pathogenic bacteria. The LAB isolate was characterized by a clear zone formed on MRSA medium + CaCO3 0.2% (w/v) and further identified by morphological and biochemical assays. The selected isolates were evaluated for their viability in low pH, pathogenic bacterial inhibition, and lactic acid production. The experimental arrangement was a factorial block design (4 x 2) consisted of four isolates and two levels of pH value (pH 2 and 3), each treatment in 3 equal replicates. The result showed that four isolates (two isolates from the rumen liquor of fistulated cattle and two isolates from silage) were identified as lactic acid bacteria. The four isolates showed inhibition activity against Escherichia coli, Bacillus subtilis, Staphylococcus aureus and performed viability at low pH during 2 h treatment. The highest lactic acid production was obtained from isolates Sil.3 (21.42%) and followed by CR2 (19.88%), CR1 (15.40%) and Sil.9 (15.08%). Biochemical identification by standard of analytical profile index (API) 50 CHL kit showed that the selected isolates CR1 was Lactobacillus paracasei ssp. paracasei 3 (91.5%), L. paracasei ssp. paracasei 3 (76.5%), Sil.3 was Lactobacillus brevis (95.1%), and Sil.9 was Lactobacillus collinoides (92.5%). In conclusion, probiotic candidates isolated from rumen liquor are confirmed as L. paracasei ssp. paracasei (CR1 and CR2), while two other isolates from king grass silage are identified as L. brevis (Sil.3) and L. collinoides (Sil.9). L. brevis (Sil.3) and L. paracasei ssp. paracasei (CR1) has higher inhibition against pathogenic bacteria (E. coli, S. aureus, and B. subtilis) than L. paracasei ssp. paracasei (CR2) and L. collinoides (Sil.9)

The development of novel silage inoculants using strain selection and genetic manipulation techniques

Eighteen strains of Pediococcus acidilactici were screened in the laboratory for their potential as silage inoculants. The grass silage isolate, P. acidilactici G24, was found to be the most suitable, primarily on the basis of its short lag phase. Inoculation of grass of water soluble carbohydrate content greater than lOOg/kg DM with P. acidilactici G24 at a rate of 106/g forage consistently aided silage preservation, as indicated by a more rapid rate of lactic acid production and pH decrease, and silage with a lower pH, a lower ammonia nitrogen content and a higher level of crude protein than uninoculated controls. Furthermore, when compared to inoculants consisting of Lactobacillus plantarum or mixtures of L. plantarum and P. acidilactici, P. acidilactici G24 initiated a more rapid rate of pH decrease and maintained the same silage pH up to 60 days after ensilage, implying that P. acidilactici G24 is superior to these more traditional inoculants. When tested on grass of water soluble carbohydrate content less than 60g/kg DM, P. acidilactici G24 failed to aid preservation. A second inoculant was therefore developed to cope with low water soluble carbohydrate conditions where starch is available as an alternative source of water soluble carbohydrate. This situation is frequently encountered in the ensilage of alfalfa. While no amylolytic strain of lactic acid bacteria suitable as a silage inoculant could be isolated, the amylase enzyme of Lactobacillus amylovorus was found to be well adapted to typical ensiling conditions. A region of DNA coding for the L. amylovorus a-amylase enzyme was therefore cloned and integrated into the Cbh gene of the competitive inoculant strain, L. plantarum LP80. A Lactobacillus host strain was chosen in order to comply with regulations governing the release of genetically engineered organisms into the environment. The amylolytic derivative, L. plantarum LPGAFA5, secreted 49% of the amylase secreted by L. amylovorus, stably maintained amylase activity for 50 generations under non-selective conditions and displayed no reduction in growth rates on glucose based media compared to the parent strain, L. plantarum LP80

Effect of Pediococcus acidilactici on Fermentation Quality, Aerobic Stability and in Vitro Digestibility of Fodder Pea, Barley, and Mixed Silages

This study aimed to evaluate the effects of Pediococcus acidilactici MF098795 strain (1×109 CFU g-1) as a silage inoculant on the fermentation characteristics, microbial population, in vitro digestibility, and aerobic stability of fodder pea (FP), barley (B), and fodder pea-barley mixture (FPB) silages. Six treatment groups were designed: control fodder pea (CFP), control barley (CB), control mixture (CFPB), and inoculated groups FPPA9, BPA9, and FPBPA9. Results indicated that inoculation with P. acidilactici improved the fermentation quality by lowering pH values and increasing lactic acid bacteria (LAB) counts, while reducing yeast and mold populations, particularly in BPA9 and FPBPA9 groups. Aerobic stability parameters, including pH₂ and CO₂ levels, were significantly reduced in the inoculated silages, suggesting enhanced resistance to aerobic spoilage. In vitro digestibility metrics such as organic matter digestibility (OMD), metabolizable energy based on gas production (MEGP), and ME based on OMD (MEOMD) were positively influenced by the inoculant. The highest improvements were observed in FPPA9 and BPA9 groups. In conclusion, the application of Pediococcus acidilactici MF098795 strain at 1×109 CFU g-1 effectively enhanced fermentation efficiency, microbial stability, and digestibility of silages, supporting its potential as a promising biological additive in forage preservation

The role of homofermentative and heterofermentative lactic acid bacteria for alfalfa silage: a meta-analysis

Lactic acid bacteria (LAB) are usually employed as alfalfa silage inoculants to obtain high-quality feed for animal husbandry. However, the effects of these inoculants are still unclear and need to be studied extensively. Therefore, the objective of this meta-analysis was to quantitatively summarize published research studies that assess the effects of homofermentative (HoLAB) and heterofermentative lactic acid bacteria (HeLAB) on fermentation parameters, nutritive value, microbiological composition and aerobic stability of alfalfa silage. PubMed, ScienceDirect and Scopus have been screened for articles published from 1980 to 2018. The criteria for inclusion were: randomized and controlled trials using alfalfa silage and published in peer-reviewed journals. It was found that inoculation with LAB decreased silage pH, neutral detergent fibre, acid detergent fibre and ammoniacal nitrogen, while it increased dry matter and crude protein compared to control in the pooled raw mean difference random-effect model. Additionally, LAB inoculation decreased acetate, propionate, ethanol and butyrate concentrations, whereas it increased lactate. In addition, inoculants reduced the counts of yeasts and moulds. Lastly, LAB inoculation improved aerobic stability. To the best of our knowledge, this is the first meta-analysis that aims at comparing the application of HoLAB and HeLAB for alfalfa silage. In the pool estimate, positive effects attributable to the application of microbial silage inoculants were found in most of the evaluated parameters; supporting the importance of applying both types of inoculants to improve forage preservation for the livestock industry.EEA RafaelaFil: Blajman, Jesica E. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Blajman, Jesica E. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; ArgentinaFil: Vinderola, Celso Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Lactologia Industrial; ArgentinaFil: Vinderola, Celso Gabriel. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactologia Industrial; ArgentinaFil: Paez, Roxana Beatriz. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; ArgentinaFil: Signorini, Marcelo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; ArgentinaFil: Signorini, Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Epiphytic microbiota in Swedish grass-clover herbage and the effect of silage additives on fermentation profiles and bacterial community compositions of the resulting silages

Aims To investigate the epiphytic microbiota in grass-clover herbage harvested at different sites and occasions and to explore the effect of different silage additives on the resulting silage microbiota.Methods and results Herbage was harvested from grass-clover leys at geographically distributed sites in a long-term field experiment in Sweden, in early and late season of two consecutive years. Different silages were made from the herbage using: (1) no additive, (2) acid-treatment, and (3) inoculation by starter culture. Herbages were analysed for botanical and chemical composition, and the resulting silages for products of fermentation. Bacterial DNA was extracted from herbage and silage samples, followed by sequencing using Illumina 16S rRNA amplicon sequencing. Herbage microbiota showed no clear correlation to site or harvesting time. Silage additives had a major effect on the ensiling process; inoculation resulted in well fermented silages comprising a homogenous microbiota dominated by the genera Lactobacillus and Pediococcus. A minor effect of harvest time was also observed, with generally a more diverse microbiota in second-harvest silages. Untreated silages showed a higher relative abundance (RA) from non-lactic acid bacteria compared to acid-treated silages. In most silages, only a few bacterial amplicon sequence variants contributed to most of the RA.Conclusions The epiphytic microbiota in grass-clover herbage were found to be random and not dependent on site. From a microbial point of view, the most predictable and preferable silage outcome was obtained by inoculation with a starter culture. Acid-treatment with formic- and propionic acid surprisingly resulted in a less preferable silage. Silage making without additives cannot be recommended based on our results

Survey About the Use of Bacterial Inoculants in Brazil: Effects on Silage Quality and Animal Performance

Our objective was to report the effect of bacterial inoculants on silage quality and animal responses in Brazil. A survey of bacterial inoculants utilization in Brazil was made based on a total of 178 published articles assessing a widely varied crops (alfalfa, cabbage, cassava, corn, grass, high-moisture corn (HMC), high-moisture sorghum, millet, oat, orange bagasse, peanut forage, sorghum, soybean, stylosantes Campo Grande, sugarcane, and sunflower). Sugarcane and grass silages comprised 58.1% of the total crops investigated. Homolactic inoculation reduced dry matter (DM) losses in alfalfa silages, but not in corn, grass, HMC, and sorghum silages. Heterolactic inoculation enhanced the aerobic stability of corn and HMC silages. The use of heterofermentative lactic acid-bacteria (LAB) was more effective to improve fermentation of sugarcane silages compared to homofermentative LAB. Inoculation impaired the DM intake in cattle fed corn, grass, and sugarcane silages, but DM intake increased in sheep due to inoculation. In some cases, silage digestibility was affected by inoculation. Positive responses to inoculation occurred most often when the compatibility between the bacterial inoculant and crop was better understood (e.g., homolactic inoculation for grass silage and heterolactic inoculation for sugarcane silage). The performance of animals consuming inoculated silages has been investigated in Brazil only a few times, but the data suggest a greater impact of bacterial inoculants on DM intake and weight gain in cattle and sheep than that indicated in temperate conditions