Reducing transmission of high-risk antibiotic resistance genes in whole-crop corn silage through lactic acid bacteria inoculation and increasing ensiling temperature.

The microbial hosts of antibiotic resistance genes (ARGs) found epiphytically on plant materials could grow and flourish during silage fermentation. This study employed metagenomic analysis and elucidated the occurrence and transmission mechanisms of ARGs and their microbial hosts in whole-crop corn silage inoculated with homofermentative strain Lactiplantibacillus plantarum or heterofermentative strain Lentilactobacillus buchneri ensiled under different temperature (20 and 30 °C). The results revealed that the corn silage was dominated by Lactobacillus, Leuconostoc, Lentilactobacillus, and Latilactobacillus. Both the ensiling temperature and inoculation had greatly modified the silage microbiota. However, regardless of the ensiling temperature, L. buchneri had significantly higher ARGs, while it only exhibited significantly higher mobile genetic elements (MGEs) in low temperature treatments. The microbial community of the corn silage hosted highly diverse form of ARGs, which were primarily MacB, RanA, bcrA, msbA, TetA (58), and TetT and mainly corresponded to macrolides and tetracyclines drug classes. Plasmids were identified as the most abundant MGEs with significant correlation with some high-risk ARGs (tetM, TolC, mdtH, and NorA), and their abundances have been reduced by ensiling process. Furthermore, higher temperature and L. buchneri reduced abundances of high-risk ARGs by modifying their hosts and reduced their transmission in the silage. Therefore, ensiling, L. buchneri inoculation and higher storage temperature could improve the biosafety of corn silage.

Harvest Timing of Standing Corn Using Near-Infrared Reflectance Spectroscopy.

Harvesting corn at the proper maturity is important for managing its nutritive value as livestock feed. Standing whole-plant moisture content is commonly utilized as a surrogate for corn maturity. However, sampling whole plants is time consuming and requires equipment not commonly found on farms. This study evaluated three methods of estimating standing moisture content. The most convenient and accurate approach involved predicting ear moisture using handheld near-infrared reflectance spectrometers and applying a previously established relationship to estimate whole-plant moisture from the ear moisture. The ear moisture model was developed using a partial least squares regression model in the 2021 growing season utilizing reference data from 610 corn plants. Ear moisture contents ranged from 26 to 80 %w.b., corresponding to a whole-plant moisture range of 55 to 81 %w.b. The model was evaluated with a validation dataset of 330 plants collected in a subsequent growing year. The model could predict whole-plant moisture in 2022 plants with a standard error of prediction of 2.7 and an R2P of 0.88. Additionally, the transfer of calibrations between three spectrometers was evaluated. This revealed significant spectrometer-to-spectrometer differences that could be mitigated by including more than one spectrometer in the calibration dataset. While this result shows promise for the method, further work should be conducted to establish calibration stability in a larger geographical region.

Effects of Whole-Plant Corn Silage on Growth Performance, Serum Biochemical Indices, and Fecal Microorganisms in Hezuo Pigs.

In this study, we investigated the effects of the dietary inclusion of different proportions of whole-plant corn silage on growth performance, serum biochemical indexes, and intestinal microorganisms in Hezuo pigs. Thirty-two two-month-old Hezuo pigs (body weight: 7.88 ± 0.81 kg) were randomly divided into four groups of eight pigs (half male, half female) each. The control (CON) group received a basal diet, while the three experimental groups were fed the basal diet, part of which had been replaced with 5%, 10%, and 15% whole-plant corn silage, respectively. The experiment lasted for 127 days, including 7 days of pre-testing and 120 days of formal testing. At the end of the experiment, blood and fecal samples were collected. Compared with the CON group, the feed-to-gain ratio was significantly lower in the 10% test group (p < 0.05), whereas the total protein, albumin, triglyceride, and glucose contents were significantly higher (p < 0.05). No significant differences in total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, creatinine, urea, aspartate aminotransferase, and alanine aminotransferase were observed among the groups (p > 0.05). The addition of whole-plant corn silage to the diet significantly increased alpha diversity in the pig gut based on 16S rRNA gene sequencing. The principal coordinate analysis results showed significant clustering of the different groups (p < 0.05). At the phylum level, the addition of whole-plant corn silage to the diet significantly decreased (p < 0.05) the relative abundance of Firmicutes and significantly increased (p < 0.05) that of Bacteroidetes. At the genus level, the relative abundance of Streptococcus significantly decreased (p < 0.05) with increasing silage supplementation levels, whereas species diversity significantly increased (p < 0.05). In conclusion, 10% is the recommended inclusion ratio for whole-plant corn silage in the diets of pigs.

Partitioning of nutrient variation in alfalfa and corn silage by source on New York dairy farms.

Variation in feed components contributes to variation and uncertainty of diets delivered to dairy cows. Forages often have a high inclusion rate (50 to 70% of DM fed) and variable composition, thus are an important contributor to nutrient variability of delivered diets. Our objective was to quantify the variation and identify the main sources of variability in corn silage and alfalfa-grass haylage composition at harvest (fresh forage) and feed-out (fermented forage) on NY dairy farms. Corn silage and alfalfa-grass haylage were sampled on 8 NY commercial dairy farms during harvest in the summer and fall of 2020 and during their subsequent feed-out in the winter and spring of 2021. At harvest, a composite sample of fresh chopped forage of every 8-ha section of individual fields was collected from piles delivered for silo filling. During a 16-week feed-out period, 2 independent samples of each forage were collected 3 times per week. The fields-of-origin of each forage sample during feed-out were identified and recorded using silo maps created at filling. A mixed-model analysis quantified the variance of corn silage DM, NDF, and starch and haylage DM, NDF, and CP content. Fixed effects included soil type, weather conditions, and management practices during harvest and feed-out while random effects were farm, silo unit, field, and day. At harvest, between-farm variability was the largest source of variation for both corn silage and haylage, but within-farm sources of variation exceeded farm-to-farm variation for haylage at feed-out. At feed-out, haylage DM and NDF content had higher within-farm variability than corn silage. In contrast, corn silage starch showed higher within-farm variation at feed-out than haylage CP content. For DM content at feed-out, day-to-day variation was the most relevant source of within-farm variation for both forages. However, for the nutrient components at feed-out (NDF and CP for haylage; NDF and starch for corn silage) silo-to-silo variation was the largest source of variability. Weather conditions systematically explained a proportion of the farm-to-farm variability for both forages at harvest and feed-out. We concluded that because of the high farm-to-farm variation, corn silage and haylage must be sampled on individual farms. We also concluded that due to the high silo-to-silo variability, and the still significant day-to-day and field-to-field variability within-farm, corn silage and haylage should be sampled within individual silos to better capture changes in forage components at feed-out.

Impact of Supplementing a Backgrounding Diet with Nonprotein Nitrogen on In Vitro Methane Production, Nutrient Digestibility, and Steer Performance.

Two experiments were conducted to evaluate the effect of nonprotein nitrogen (NPN) supplementation on in vitro fermentation and animal performance using a backgrounding diet. In experiment 1, incubations were conducted on three separate days (replicates). Treatments were control (CTL, without NPN), urea (U), urea-biuret (UB), and urea-biuret-nitrate (UBN) mixtures. Except for control, treatments were isonitrogenous using 1% U inclusion as a reference. Ruminal fluid was collected from two Angus-crossbred steers fed a backgrounding diet plus 100 g of a UBN mixture for at least 35 d. The concentration of volatile fatty acids (VFA) and ammonia nitrogen (NH3-N), in vitro organic matter digestibility (IVOMD), and total gas and methane (CH4) production were determined at 24 h of incubation. In experiment 2, 72 Angus-crossbred yearling steers (303 ±â€…29 kg of body weight [BW]) were stratified by BW and randomly allocated in nine pens (eight animals/pen and three pens/treatment). Steers consumed a backgrounding diet formulated to match the diet used in the in vitro fermentation experiment. Treatments were U, UB, and UBN and were isonitrogenous using 1% U inclusion as a reference. Steers were adapted to the NPN supplementation for 17 d. Then, digestibility evaluation was performed after 13 d of full NPN supplementation for 4 d using 36 steers (12 steers/treatment). After that, steer performance was evaluated for 56 d (24 steers/treatment). In experiment 1, NPN supplementation increased the concentration of NH3-N and VFA (P < 0.01) without affecting the IVOMD (P = 0.48), total gas (P = 0.51), and CH4 production (P = 0.57). Additionally, in vitro fermentation parameters did not differ (P > 0.05) among NPN sources. In experiment 2, NPN supplementation did not change dry matter and nutrient intake (P > 0.05). However, UB and UBN showed lower (P < 0.05) nutrient digestibility than U, except for starch (P = 0.20). Dry matter intake (P = 0.28), average daily gain (P = 0.88), and gain:feed (P = 0.63) did not differ among steers receiving NPN mixtures. In conclusion, tested NPN mixtures have the potential to be included in the backgrounding diets without any apparent negative effects on animal performance and warrant further studies to evaluate other variables to fully assess the response of feeding these novel NPN mixtures.

Nonprotein nitrogen (NPN) supplements can be used as a nitrogen source for ruminants fed low-protein diets. The most common NPN source is urea, included typically at a range between 0.5% and 1% of the diet dry matter in growing beef cattle. Although other NPN sources and mixtures are available, there is scarce information regarding their use in ruminant production. Two experiments were conducted to evaluate the effect of NPN sources on in vitro fermentation and animal performance using a backgrounding diet. In experiment 1, three different incubations were performed for 24 h. Treatments were control (without NPN), urea (U), urea­biuret (UB), and urea­biuret­nitrate (UBN) mixtures. In experiment 2, 72 crossbred yearling steers were randomly assigned to one of the following treatments: U, UB, and UBN mixtures. Diets were formulated to contain the same nitrogen concentration in both experiments. In experiment 1, supplementation of NPN increased the in vitro fermentation, but there were no differences among NPN sources. In experiment 2, steers performed similarly among NPN sources. These findings suggest that NPN mixtures have the potential to be included in the backgrounding diets without detrimental effects. Further studies should evaluate other variables (e.g., fermentation dynamic and microbial protein supply) when using these novel mixtures.

Nutrient intake, productive and metabolic parameters of Nellore bulls feed a forage free diet using cottonseed cake as a fiber source.

We hypothesized that cottonseed cake in confined Nellore young bulls' diet as fiber source, could maintain or improve the nutrient intake, and productive and metabolic parameters. It was evaluated the total replacement of whole plant corn silage (WPCS) by cottonseed cake (CSC) as a source of fiber in the diet. A completely randomized design with two treatments and 12 replications was used. The treatments were two experimental diets containing 300 g/kg of WPCS or CSC as roughage. All animals at 0, 15, 30, 60 and 112 days were weighed to monitor productive performance and nutrient intake. Dry matter intakes, organic matter, neutral detergent fiber, physically effective neutral detergent fiber, crude protein and non-fiber carbohydrates of the WPCS treatment cattle were higher (p < 0.05), and the ethereal extract intake of cattle fed CSC diet was higher. The BW of cattle fed WPCS was higher in the two initial periods of confinement (p < 0.05). Animals from both treatments showed linear growth during the confinement period. There was a fiber source × period interaction for total weight gain (TWG), average daily gain (ADG) and feed efficiency (FE) (p < 0.05), with the lowest in the first 15 days and the highest at 30 days, reflecting a compensatory gain in animals fed with CSC. Bulls fed with CSC showed negative TWG and ADG in the first period (p < 0.05), but recovered in the following periods, surpassing that of the SC treatment in the last two periods. There was negative EF of bulls fed with CSC in the first period (P < 0.05), but in the following periods the CSC diet did not differ from the WPCS diet. The animals fed CSC diet showed a higher concentration of cholesterol and glucose in the blood plasma (p < 0.05). The forage-free diet containing cottonseed cake as a source of fiber replacing corn silage promoted better animal performance.

Effect of feeding corn silage on semen quality and spermatogenesis of bulls.

Bovine reproduction, including male fertility traits like semen quality, are influenced by a variety of different factors like breed, nutrition, environment, and feeding management. Diet in a crucial determinant, and in this regard although corn silage is generally considered to be a favorable roughage for fattening meat type breeds, it tends to have a negative impact on semen quality. In the current study, alfalfa hay was substituted by corn silage as a roughage source in the diet of bulls to investigate its effects on the fertility of breeding bulls. A feeding trail spanning 140 days was conducted, with semen collection occurring twice a week commencing 60 days after the start of trial. Semen quality parameters, serum antioxidant indexes, sex hormone content in semen, rumen microflora, and sperm transcriptome were characterized. Feeding corn silage enhanced host antioxidant capacity, significantly decreased spermatozoal motility and increased sperm deformity rate in bulls. Furthermore, follicle-stimulating hormone (FSH) and luteinizing hormone (LH) content in semen were significantly decreased (P < 0.05), and the inhibin B (INHB) content was significantly increased (P < 0.01). Feeding corn silage led to significant changes in the diversity of rumen microbiota of cattle at the phylum and genus levels, some of which were significantly correlated with semen quality. Subsequent RNA sequencing indicated that DHH and PITHD1, two genes related to sperm and reproductive development, were differentially expressed, and enrichment analysis also identified several pathways and biological functions relevant to sperm development and reproduction. These results indicate that feeding corn silage modulates semen quality via different pathways. Firstly, corn silage metabolites likely affect the secretion of INHB through the testicular capillaries, which affects semen quality by regulating genes involved in spermatogenesis. Secondly, low lignin content in silage corn appears to reduce abundance of rumen flora that are positively correlated with semen quality. Overall, results indicate that feeding bulls corn silage as the primary source of forage could negatively impact semen quality and may not be appropriate as the primary roughage of forage for breeding bulls.

Effects of feeding different levels of dietary corn silage on growth performance, rumen fermentation and bacterial community of post-weaning dairy calves.

OBJECTIVE: The objective of this study was to evaluate the growth performance, rumen fermentation parameters and bacterial community of post-weaning dairy calves in response to five diets varying in corn silage (CS) inclusion. METHODS: A total of forty Holstein weaned bull calves (80±3 days of age;128.2±5.03 kg at study initiation) were randomized into five groups (8 calves/group) with each receiving one of five dietary treatments offered as total mixed ration in a 123-d feeding study. Dietary treatments were control diet (CON; 0% CS dry matter [DM]); Treatment 1 (T1; 27.2% CS DM); Treatment 2 (T2; 46.5% CS DM); Treatment 3 (T3; 54.8% CS DM); and Treatment 4 (T4; 67.2% CS DM) with all diets balanced for similar protein and energy concentration. RESULTS: Results showed that calves offered CS had greater average daily gain, body length and chest depth growth, meanwhile altered rumen fermentation indicated by decreased rumen acetate concentrations. Principal coordinate analysis showed the rumen bacterial community structure was affected by varying CS inclusion diets. Bacteroidetes and Firmicutes were the predominant bacterial phyla in the calf rumens across all treatments. At the genus level, the abundance of Bacteroidales_RF16_group was increased, whereas Unclassified_ Lachnospiraceae was decreased for calves fed CS. Furthermore, Spearman's correlation test between the rumen bacteria and rumen fermentation parameters indicated that Bacteroidales_RF16_group and Unclassified Lachnospiraceae were positively correlated with propionate and acetate, respectively. CONCLUSION: The results of the current study suggested that diet CS inclusion was beneficial for post-weaning dairy calf growth, with 27.2% to 46.5% CS of diet DM recommended to achieve improved growth performance. Bacteroidales_RF16_group and Unclassified Lachnospiraceae play an important role in the rumen fermentation pattern for post-weaning calves fed CS.

Impact of corn silage substitution for dry alfalfa on milk fatty acid profile, nitrogen utilization, plasma biochemical markers, rumen fermentation, and antioxidant capacity in Mahabadi lactating goats.

This study aimed to investigate the effects of substituting conventional dry forage with corn silage, a more cost-effective supplementary feed, on a variety of parameters in lactating Mahabadi goats, native to arid and semi-arid regions. A total of fifty lactating Mahabadi goats, aged between 2 and 5 years and weighing an average of 45.3 ± 7.20 kg, were randomly allocated into two experimental groups. The experimental treatments consisted of a control diet and a diet incorporating corn silage at 20 % of dry matter (DM). Various measurements were conducted to assess milk compositions, fatty acid profiles, rumen fermentation parameters, selected plasma biochemical parameters, nitrogen balance, and antioxidant capacity. The results revealed that the substitution of dry forage with corn silage had no statistically significant impact on milk yield, dry matter intake (DMI), body weight, as well as the proportions of monounsaturated and unsaturated fatty acids, and specifically C18:1C9 in milk, non-esterified fatty acid content in milk, rumen concentration of volatile fatty acids (VFA), microbial nitrogen, and nitrogen balance (p > 0.05). Nonetheless, this substitution led to a reduction in the percentage of milk polyunsaturated fatty acids and plasma urea nitrogen content (p < 0.05). It is noteworthy that goats fed with corn silage exhibited a notably higher antioxidant capacity in rumen liquor (p < 0.05), plasma, and milk (p < 0.01) compared to those receiving the control diet. The substitution of conventional dry forage with corn silage exhibited no adverse effects on the performance of lactating Mahabadi goats. On the contrary, it contributed to an enhancement in the antioxidant status. These findings offer valuable insights for optimizing dietary composition by incorporating corn silage at 20 % of DM, thereby improving milk quality, reducing costs, and promoting overall health in lactating goats, particularly within arid and semi-arid regions.

Optimizing corn silage quality during hot summer conditions of the tropics: investigating the effect of additives on in-silo fermentation characteristics, nutrient profiles, digestibility and post-ensiling stability.

Corn crop grown and ensiled at high temperature have lower water soluble carbohydrates (WSC), epiphytic lactic acid bacteria (LAB) population, lactic acid concentration, fermentation quality and aerobic stability. This study systematically investigated the effects of heterofermentative LAB (hetLAB), homofermentative LAB (homLAB), molasses and their mixture (MIX) on in-silo fermentation characteristics, chemical profiles, Cornell Net Carbohydrate and Protein System (CNCPS) carbohydrate subfractions, in vitro digestibility (DMD), microbial count, and post-ensiling aerobic stability of whole crop corn silage during hot summer (30 to 45°C) condition. Corn hybrids (P30K08 and DK6789) were ensiled at targeted dry matter (DM) of 330 g/kg for 0, 3, 7, 21, and 150 days in 3 L silos, without additive (CCS) or treated with hetLAB (4×106 cfu/g Lactobacillus buchneri), homLAB (1×106 cfu/g of L. plantarum), molasses (3% of fresh forage) or MIX (half of individual doses of homLAB, hetLAB and molasses) additives. The CCS, homLAB, hetLAB, molasses, or MIX treated chopped material of each hybrid were ensiled in 16 replicate silos at a density of 260 kg of DM/m3. Compared to CCS, the additives significantly improved silage nutritional and fermentation quality, DM digestibility (in vitro), count of LAB, DM recovery and aerobic stability, and decreased counts of yeast and mold. Among the inoculants, the homLAB and MIX inoculated silages had greatest improvement in fermentation quality and nutritional value. The homLAB produced corn silage with the highest (P < 0.05) content of lactic acid, and soluble carbohydrates, and lowest contents of acetic acid, NH3-N and pH, demonstrating desirable and restricted in silo fermentation. On the other hand, the hetLAB inoculated silages had the greatest (P < 0.05) value of acetic acids, highlighting greater aerobic stability. Interestingly, the MIX silages followed the hetLAB in acetic acid value and homLAB in lactic acid value. Notably, without additive stable pH was not achieved during 21 days, with application of molasses, hetLAB and the MIX inoculants stable pH was achieved during 7 days, and with homLAB stable pH was achieved during the first 3 days of ensiling. The greatest numbers of viable LAB were recorded in homLAB (8.13 log cfu/g) and MIX (7.89 log cfu/g) inoculated silages, while the lowest for CCS (6.29 log cfu/g). The lowest yeast (1.48 log cfu/g) and mold (0.22 log cfu/g) were recorded for hetLAB inoculated silage. The greatest (P < 0.05) DM recovery was recorded for hetLAB (97.3%) and MIX (96.9%), and the lowest for the control silage (92.9%). All additives significantly improved the aerobic stability of corn silage, and the greatest value of >72 h was recorded for hetLAB and MIX inoculats, and the lowest for CSC (25 h). In conclusion, additives application can improve fermentation quality, nutritional value, DM recovery and aerobic stability of whole crop corn silage under hot summer conditions of the tropics. The MIX inoculant showed potential to improve in-silo fermentation, and aerobic stability at the same time, however, further investigation are required, particularly with respect of dose rate.

Hazardous risk of antibiotic resistance genes: Host occurrence, distribution, mobility and vertical transmission from different environments to corn silage.

Antibiotic resistance genes (ARGs) are one of the emerging contaminants posing a great deal of hazardous risk to public health. This study employed metagenomics and deciphered the potential risk of the antibiotic resistome and their vertical transfer to ensiled whole-crop corn silage harvested from six climate zones: 1. Warm temperate-fully humid-hot summer (Cfa), 2. Arid-desert-cold arid (BWk), 3. Snow-desert-cold summer (Dwc), 4. Snow-desert-hot summer (Dwa), 5. Arid-steppe-cold arid (BSk), and 6. Equatorial-desert (Aw) based on the Köppen-Geiger climate classification in China. The findings demonstrate a high diversity of ARGs, which is related to the drug classes of tetracycline, ciprofloxacin, lincosamide, fosfomycin, and beta lactam. Resistome variations are mostly related to variations in microbial composition and fermentation characteristics of the silages from different climate zones, which are indirectly influenced by environmental conditions. The most dominating ARGs in corn silage were tetM, acrA, H-NS, lnuA, emrR, and KpnG, which is primarily hosted by Klebsiella and Lactobacilli. There were 5 high-risk ARGs (tetM, bacA, SHV-1, dfrA17, and QnrS1) in silage from different climate zones, and the tetM was the most prevalent high-risk ARG. However, throughout the ensiling process, the abundance of ARGs, and mobile ARGs were reduced. The resistome contamination in silage from Tibet (Dwc) with high altitude and harsh environment was relatively low due to the low variety and abundance of ARGs, the low abundance of mobile ARGs and high-risk ARGs. In addition, most of the bacteria responsible for the silage fermentation were also found to be the hosts to the ARGs, although their abundance decreased after 90 d of silage fermentation. Hence, we alert the existence of ARGs-related biosafety risk in silages and call for more attention to the silage ARGs, their hosts, and mobile genetic elements in order to curtail their possible risk to public health.

Effects of fermentative quality of corn silage on lactation, blood metabolites, and rumen fermentation in dairy cows.

We investigated the effect of fermentation quality of corn silage on dry matter intake (DMI), milk yield, ruminal fermentation, methane (CH4 ) emissions, and plasma metabolites in lactating cows. Six lactating Holstein cows were used in a 2 × 2 crossover design with two dietary treatments containing high quality corn silage with lower pH (high group) or low quality corn silage with higher pH (low group). The cows were fed partial mixed ration (PMR containing 50%DM of each corn silage) ad libitum plus 0.7 kg/day of concentrates at milking. The DMI of cows in the low group (24.8 kg/day) tended to be lower (p < 0.10) than that in the high group (26.8 kg/day). The dietary treatment did not affect milk yield or milk fat, protein, or lactose concentrations. The ruminal acetic acid proportion of the low group was significantly higher (p < 0.05) than that of the high group. The CH4 emission per DMI of the low group tended to be higher (p < 0.10) than that of the high group. The plasma concentration of the total cholesterol (TCHO) and the activity of aspartate aminotransferase and alanine aminotransferase of the low group were significantly higher than those of the high group.

Responses of rumen microorganisms and metabolites to different roughage of domesticated Tibetan sheep.

Tibetan sheep can utilize high fiber feeds well. However, the mechanisms of rumen microbiota and metabolites in response to different roughage in a housed environment are still unclear. We fed Tibetan sheep with three different roughage diets: 50% whole corn silage (TS), 50% wheatgrass group (TW), and 25% each of whole corn silage and wheatgrass (TM). Subsequently, meat traits, rumen contents 16S rRNA and metabolomics were studied. The results showed that feeding wheat straw to Tibetan sheep significantly increased the abundance of bacteria such as Ruminococcus and Succiniclasticum in the rumen. These microorganisms significantly increased metabolites such as beta-alanyl-L-lysine, butanoic acid and prostaglandin E2. Eventually, production performance, such as carcass weight and intramuscular fat and meat quality characteristics, such as color and tenderness were improved by altering the rumen's amino acid, lipid and carbohydrate metabolism. This study demonstrated that including 25% wheatgrass and 25% whole corn silage in the diet improved the performance of Tibetan sheep, revealing the effect of the diet on the performance of Tibetan sheep through rumen microorganisms and metabolites.

Comprehensive profiling of the metabolome in corn silage inoculated with or without Lactiplantibacillus plantarum using different untargeted metabolomics analyses.

Silage fermentation is a complicated biochemical process involving interactions between microbes and metabolites. However, the overall metabolome feature of ensiled forage and its response to lactic acid bacteria inoculation is poorly understood. Hence, in this study metabolome profiles of whole-plant corn silage inoculated with or without Lactiplantibacillus plantarum were characterised via solid-phase microextraction/gas chromatography/mass spectrometry (SPME-GC-MS), gas chromatography/time-of-flight mass spectrometry (GC-TOF-MS), and Liquid chromatography/Q Exactive HFX mass spectrometry (LC-QE-MS/MS) analysis. There were 2087 identified metabolites including 1143 reliably identified metabolites in fresh and ensiled whole-plant corn. After ensiling, the increased metabolites in whole-plant corn were mainly composed of organic acids, volatile organic compounds (VOC), benzene and substituted derivatives, carboxylic acids and derivatives, fatty acyls, flavonoids, indoles and derivatives, organooxygen compounds (including amines and amides), phenols, pyridines and derivatives, and steroids and steroid derivatives, which includes neurotransmitters and metabolites with aromatic, antioxidant, anti-inflammatory, and antimicrobial activities. Phenylacetaldehyde was the most abundant aromatic metabolite after ensiling. L-isoleucine and oxoproline were the major free amino acids in silage. Ensiling markedly increased the relative abundances of 3-phenyllactic acid, chrysoeriol, 6-O-acetylaustroinulin, acetylcholine, γ-aminobutyric acid, pyridoxine, and alpha-linoleic acid. Inoculation with L. plantarum remarkably changed silage VOC composition, and essential amino acids, 3-phenyllactic acid, and cinnamaldehyde compared with untreated silage. The present study does not only provide a deeper insight into metabolites of the ensiled whole-plant corn but also reveals metabolites with specific biological functions that could be much helpful in screening novel lactic acid bacteria to well ensile forages. Inoculation with L. plantarum significantly affects the metabolome in ensiled whole-plant corn.

Flint corn silage management: influence of maturity stage, inoculation with Lentilactobacillus buchneri, and storage time on fermentation pattern, aerobic stability, and nutritional characteristics.

Introduction: High quality corn silage depends on factors such as corn type, stage of crop development at harvest time, fermentation time, in addition to use or not of inoculants. This study aimed to investigate the impact of maturity stage, bacterial inoculation, and storage time on fermentation, aerobic stability, and nutritional characteristics of flint corn silage and their implications for corn silage management. Methods: A flint corn hybrid was harvested very early, early, and medium (at 250, 300 and 350 g dry matter (DM)/kg as fed, respectively) and ensiled in mini-silos without (control) or with Lentilactobacillus buchneri CNCM I-4323 at 1 × 105 cfu/g for 120, 240 and 360 d to investigate how these factors interact with each other. Results and discussion: There was only a small increase (7 g/kg starch; p = 0.003) in starch digestibility (starch-D) in the silages stored for 360 d when compared to that stored for 240 d, but with no difference for 120 d. Despite the reduced starch-D (526 vs. 694 g/kg starch; p < 0.001), silages produced from medium harvest had higher (p < 0.001) starch content (317 vs. 137 g/kg DM) and higher amount of digestible starch (169 vs. 98.5 g/kg DM; p < 0.001) compared to very early harvest. The 2-way interactions (inoculation × storage time and maturity × storage time) showed that inoculation of corn silage with L. buchneri increased (p < 0.001) the aerobic stability, and that more mature crop silage had higher aerobic stability (140 h; p = 0.036) than the others (118 and 48.5 h for those silages from very early and early harvest). Conclusion: The storage for a longer time (>120 d) with the goal of increasing silage digestibility did not occur. Harvesting whole-crop flint corn with 300 to 350 g/kg DM is desirable to have higher DM yield and starch accumulation. Inoculation with L. buchneri is recommended to preserve the silage against aerobic deterioration. This study has shown the importance of harvesting flint corn at the right time, and the need for inoculation with L. buchneri to ensure greater yield, starch accumulation, and silage preservation, if 120 days of storage are not exceeded.

Effects of endosperm type and storage length of whole-plant corn silage on nitrogen fraction, fermentation products, zein profile, and starch digestibility.

Zeins are commercially important proteins found in corn endosperms. The objective of this study was to evaluate the effect of altering zein levels in corn inbred lines carrying endosperm mutations with differential allelic dosage and analyze the effects on the composition, nutritive value, and starch digestibility of whole-plant corn silage (WPCS) at 5 storage lengths. Three inbred lines carrying 3 different endosperm modifiers (opaque-2 [o2], floury-2 [fl2], and soft endosperm-1 [h1]) were pollinated with 2 pollen sources to form pairs of near-isogenic lines with either 2 or 3 doses of the mutant allele for each endosperm modifier. The experiment was designed as a split-plot design with 3 replications. Pollinated genotype was the main plot factor, and storage length was the subplot-level factor. Agronomic precautions were taken to mimic hybrid WPCS to the extent possible. Samples were collected at approximately 30% dry matter (DM) using a forage harvester and ensiled in heat-sealed plastic bags for 0, 30, 60, 120, and 240 d. Thus, the experiment consisted of 30 treatments (6 genotypes × 5 storage lengths) and 90 ensiling units (3 replications per treatment). Measurements included nutrient analysis, including crude protein, soluble crude protein, amylase-treated neutral detergent fiber, acid detergent fiber, lignin, starch, fermentation end products, zein concentration, and in vitro starch digestibility (ivSD). The nutritional profile of the inbred-based silage samples was similar to hybrid values reported in literature. Significant differences were found in fresh (unfermented) sample kernels for endosperm vitreousness and zein profiles between and within isogenic pairs. The o2 homozygous (3 doses of mutant allele) had the highest reduction in vitreousness level (74.5 to 38%) and zein concentration (6.2 to 4.7% of DM) compared with the heterozygous counterpart (2 doses of mutant allele). All genotypes showed significant reduction of total zeins and α-zeins during progressive storage length. In vitro starch digestibility increased with storage length and had significant effects of genotype and storage length but not for genotype by storage length interaction, which suggests that the storage period did not attenuate the difference in ivSD between near-isogenic pairs caused by zeins in WPCS. Both total zeins and α-zeins showed a strong negative correlation with ivSD, which agrees with the general hypothesis that the degradation of zeins increases ruminal starch degradability. Homozygous o2 was the only mutant with significantly higher ivSD compared with the heterozygous version, which suggests that, if all other conditions remain constant in a WPCS systems, substantial reductions in endosperm α-zeins are required to significantly improve ivSD in the silo.

The effect of biogas slurry application on biomass production and the silage quality of corn.

OBJECTIVE: The objective of this study was to evaluate the effect of biogas slurry application on biomass production and the silage quality of corn. METHODS: A field experiment was conducted in which corn was grown using different biogas slurry application rates. The effect of 25% to 500% biogas slurry nitrogen replacement (T1 to T14) on the yield and quality indices of corn were studied by field plot experiments. RESULTS: The results revealed that biogas slurry application improved the stem diameter and relative feed value of corn silage in treatments T13 and T11. Moreover, the fermentation quality of corn silage was improved due to an increase in lactic acid content; in comparison with the chemical synthetic fertilizer (CF) group. The crude protein contents of corn silage had no obvious change with increasing biogas slurry application. However, the forage quality index of acid detergent fiber was decreased (p<0.05) in the T11 group compared with the CF group. In addition, higher (p<0.05) 30 h in vitro dry matter digestibility and 30 h in vitro neutral detergent fiber digestibility were observed in the T11 and T13 groups than in the CF group. CONCLUSION: Based on these results, it was concluded that the optimum biogas slurry application rate for corn was approximately 350% to 450% biogas slurry nitrogen replacement under the present experimental conditions.

Whole-plant flint corn silage inclusion in total mixed rations for pre- and postweaning dairy calves.

Assuming that acetic acid plays a minor role in the development of ruminal epithelium of preweaning dairy calves, the fiber supply for growing calves has been neglected. More research has been done on including starch and nonfibrous carbohydrates in solid feed for preweaning calves. Accordingly, the fiber requirement of these calves is not well known, as diet recommendations vary greatly. Hence, elucidating the effects of including fiber from long particle sizes in the diet may be essential for helping calves overcome the transition challenge during weaning. Forty-five Holstein calves were used in a randomized block design, considering sex, birth date, and weight at 28 d of age, when the supply of the total mixed ration (TMR) with the inclusion of corn silage started. Three TMR with increasing whole-plant flint corn silage content (0, 10, or 20% on a dry matter basis) were compared: 0CS, 10CS, or 20CS, respectively. During the first 28 d of life, the calves were managed homogeneously and were fed 6 L/d of whole milk, a commercial calf starter pelleted, and water ad libitum. Next, the solid diet was changed to the respective solid feed treatment. Calves were gradually weaned from 52 to 56 d of age but were evaluated for an additional 14 d postweaning. Feed intake was measured daily, while body weight and metabolic indicators of intermediate metabolism were evaluated weekly. Ruminal fluid was collected at 6, 8, and 10 wk of age. Behavioral analysis was conducted on wk 7 (preweaning) and 10 (postweaning). There was a quadratic effect for dry matter intake from wk 7 to 10, with higher intake for the 10CS diet than the 0CS and 20CS diets. Consequently, the 10CS diet also promoted greater average daily gain at wk 8 and 9 compared with the 0CS and 20CS diets. However, the final body weight was not affected by the different solid diets. Silage inclusion in calves' diet positively affected time spent ruminating and chewing pre- and postweaning. Including 10% of whole-plant flint corn silage in the diets of young dairy calves is a strategy to increase total solid intake and decrease acidosis risk by increasing pH and ruminating activity around weaning.

Long transportation duration affects nutrient composition, mycotoxins and microbial community in whole-plant corn silage.

Introduction: Potential nutrient losses and mycotoxin accumulation caused by abnormal fermentation during transportation from cropland to dairy farms leads to the diseases incidence and threatens the health of dairy cows, then further causes financial losses. The aim of this study was to investigate the effects of different transportation times on the nutritional composition, mycotoxins, and microbial communities in whole-plant corn silage (WPCS). Methods: Three groups were subjected to different transport times: DY, short (<200 min); ZY, medium time (300-500 min); and CY, long transport time (>600 min). WPCS were collected from the same field, and nutrient composition and microbial composition before and after transportation were analyzed. Results and discussion: Our results showed that the temperature of WPCS was higher in the ZY and CY groups than in the DY group (P < 0.01). There were no significant differences in dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), ether extract (EE) and starch contents after different transportation times (P > 0.05), whereas the starch and water-soluble carbohydrates (WSC) contents in the CY group was significantly decreased after transport (P < 0.05). Similarly, the concentration of vomitoxin in the DY and CY groups declined markedly (P < 0.05) and the zearalenone content in the DY group also significantly decreased after transportation (P < 0.05). Regarding the analysis of microorganisms in WPCS, UniFrac-distance matrices and Shannon indices showed differences in the ZY group (P < 0.05), but fungal diversities were not influenced by the transport time (P > 0.05). In the ZY group, the relative abundance of Lactiplantibacillus decreased significantly after transportation (P > 0.05), but the relative abundances of unidentified_Chloroplast, Pantoea, Gluconobacter, unidentified Acetobacter and Acinetobacter increased markedly (P < 0.05). In addition, the relative abundances of Acetobacter and Gluconobacter in the CY group increased after transport (P < 0.05). Among fungal communities, a total of three, nine, and ten different fungal flora were observed in the DY, ZY, and CY groups, respectively, although no difference was found in fungal diversity. In conclusion, increased temperature, loss of starch, and mycotoxin variation were found with increased transport time. This might be the result of competition between bacteria and fungi, and novel technologies will need to be utilized for further exploration of the mechanism.

Application of different proportions of sweet sorghum silage as a substitute for corn silage in dairy cows.

This experiment explored the effects of different proportions of sweet sorghum silage as a substitute for corn silage on dry matter intake (DMI), milk yield, milk quality, apparent digestibility, rumen fermentation parameters, serum amino acid profile, and rumen microbial composition of dairy cows. A total of 32 mid-lactation Holstein dairy cows with similar body weights and parities were randomly divided into four treatments: 100% corn silage +0% sorghum silage (CON), 75% corn silage +25% sorghum silage (CS1), 50% corn silage +50% sorghum silage (CS2), and 25% corn silage +75% sorghum silage (CS3). The milk yield was increased (linear, p = .048) as the proportion of sweet sorghum increased. Linear (p = .003) and quadratic (p = .046) increased effects were observed in milk fat as corn silage was replaced with sorghum silage. Compared with the CON diet group, the CS2 and CS3 diet groups had lower dry matter (DM) (linear, p < .001), ether extract (EE) (linear, p < .001), and gross energy (GE) (linear, p = .001) digestibility of the dairy cows. The ruminal fluid aspartate (Asp) level decreased (linear, p = .003) as the proportion of sweet sorghum increased. Linear (p < .05) and quadratic (p < .05) increased effects were observed for the contents of threonine (Thr), glycine (Gly), valine (Val), leucine (Leu), tyrosine (Tyr), and histidine (His) in rumen fluid with the replacement of corn silage with sorghum silage. Cows fed the CS3 diet had greater Faecalibacterium, Bacteroides, and Prevotella ruminicola content/copy number than those fed the CON diet (p < .05). In conclusion, feeding sorghum silage as a replacement for corn silage could increase the milk yield and fat, promote the growth of rumen microbes, and provide more rumen fluid amino acids for the body and microbial utilization. We believe that sorghum silage is feasible for dairy cows, and it is reasonable to replace corn silage with 75% sorghum silage.