Lacticaseibacillus parahuelsenbergensis sp. nov., Lacticaseibacillus styriensis sp. nov. and Lacticaseibacillus zeae subsp. silagei subsp. nov., isolated from different grass and corn silage.

Four rod-shaped, non-motile, non-spore-forming, facultative anaerobic, Gram-stain-positive lactic acid bacteria, designated as EB0058T, SCR0080, LD0937T and SCR0063T, were isolated from different corn and grass silage samples. The isolated strains were characterized using a polyphasic approach and EB0058T and SCR0080 were identified as Lacticaseibacillus zeae by 16S rRNA gene sequence analysis. Based on whole-genome sequence-based characterization, EB0058T and SCR0080 were separated into a distinct clade from Lacticaseibacillus zeae DSM 20178T, together with CECT9104 and UD2202, whose genomic sequences are available from NCBI GenBank. The average nucleotide identity (ANI) values within the new subgroup are 99.9 % and the digital DNA-DNA hybridization (dDDH) values are 99.3-99.9 %, respectively. In contrast, comparison of the new subgroup with publicly available genomic sequences of L. zeae strains, including the type strain DSM 20178T, revealed dDDH values of 70.2-72.5 % and ANI values of 96.2-96.6 %. Based on their chemotaxonomic, phenotypic and phylogenetic characteristics, EB0058T and SCR0080 represent a new subspecies of L. zeae. The name Lacticaseibacillus zeae subsp. silagei subsp. nov. is proposed with the type strain EB0058T (=DSM 116376T=NCIMB 15474T). According to the results of 16S rRNA gene sequencing, LD0937T and SCR0063T are members of the Lacticaseibacillus group. The dDDH value between the isolates LD0937T and SCR0063T was 67.6 %, which is below the species threshold of 70 %, clearly showing that these two isolates belong to different species. For both strains, whole genome-sequencing revealed that the closest relatives within the Lacticaseibacillus group were Lacticaseibacillus huelsenbergensis DSM 115425 (dDDH 66.5 and 65.9 %) and Lacticaseibacillus casei DSM 20011T (dDDH 64.1 and 64.9 %). Based on the genomic, chemotaxonomic and morphological data obtained in this study, two novel species, Lacticaseibacillus parahuelsenbergensis sp. nov. and Lacticaseibacillus styriensis sp. nov. are proposed and the type strains are LD0937T (=DSM 116105T=NCIMB 15471T) and SCR0063T (=DSM 116297T=NCIMB 15473T), respectively.

Effects of protein and forage source on performance, and splanchnic and mammary net fluxes of nutrients in lactating dairy cows.

This study was conducted to determine if the decreased MP supply predicted by the NRC (2001) when canola meal (CM) substitutes soybean meal (SBM) was supported by direct measurement of net portal absorption of AA or energy-yielding nutrients, plus the impact of the type of forage in CM-based rations. Nine Holstein cows with indwelling catheters in splanchnic blood vessels, 8 also with a ruminal cannula were used to examine the effects of protein source in corn silage-based diets, comparing SBM versus CM, and forage source in CM-based diets, comparing corn versus grass silage. The cows were allocated to a triple 3 × 3 Latin square design with 21-d periods. The 3 experimental diets, formulated to be isoenergetic and isonitrogenous, were based on: 1) SBM and corn silage (SoyCorn); 2) CM and corn silage (CanCorn) and 3) CM and cool-season grass silage (CanGrass). Averages of intake, milk yield and milk composition of the last 3 d of each period were used for statistical analyses. On d 21 of each period, 6 sets of arterial, portal, hepatic and mammary blood samples and 2 ruminal fluid samples were collected. On d 12 of period 2, the protein sources were incubated in nylon bags to determine 16h-ruminal disappearance of DM and N and to obtain 16-h residues. Finally, 5 d after the completion of the Latin square design, the mobile bag technique was used to determine DM and N intestinal disappearance of the 16-h residues of SBM and CM. Pre-planned contrasts were used to compare the effect of the protein source in cows fed corn silage, i.e., SoyCorn versus CanCorn, and the effect of forage in cows fed CM, i.e., CanCorn versus CanGrass. Data of the cow without a rumen canula could not be used because of health problem. In corn silage-based diets, substitution of SBM by CM tended to increase milk (6%) and milk fat (7%) yields. The 8% higher ruminal N disappearance and the 19% decreased MP supply from RUP predicted by NRC (2001) were not supported by the 25% decrease in ruminal ammonia concentration, similar net portal absorption of AA (except 22% higher for Met), and the 14% decrease in urea hepatic removal when CM substituted SBM. Ruminal incubation of CM in nylon bags does not appear suitable for adequate determination of the rumen by-pass of a protein source like CM. Inclusion of grass silage rather than corn silage in CM-based diets tended to increase milk (6%) and increased milk lactose (8%) yields. Neither protein nor forage source resulted in variations of metabolism of energy-yielding nutrients that could explain observed increments in cow performance. The present study indicates no decreased AA availability when CM substitutes SBM. Therefore, substitution of SBM by CM in diets based on corn silage and CM in corn- or grass silage-diets can be used successfully in high producing dairy cows.

Effect of Regulation of Whole-Plant Corn Silage Inoculated with Lactobacillus buchneri or Bacillus licheniformis Regarding the Dynamics of Bacterial and Fungal Communities on Aerobic Stability.

Enhancing the aerobic stability of whole-plant corn silage is essential for producing high-quality silage. Our research assessed the effect of inoculation with Lactobacillus buchneri or Bacillus licheniformis and its modulation of the bacterial and fungal microbial community structure in an aerobic stage of whole-plant corn silage. Following treatment with a distilled sterile water control, Lactobacillus buchneri, and Bacillus licheniformis (2 × 105 cfu/g), whole-plant corn was ensiled for 60 days. Samples were taken on days 0, 3, and 7 of aerobic exposure, and the results showed that inoculation with Lactobacillus buchneri or Bacillus licheniformis improved the aerobic stability of silage when compared to the effect of the control (p < 0.05). Inoculation with Bacillus licheniformis attenuated the increase in pH value and the decrease in lactic acid in the aerobic stage (p < 0.05), reducing the filamentous fungal counts. On the other hand, inoculation with Lactobacillus buchneri or Bacillus licheniformis increased the diversity of the fungal communities (p < 0.05), complicating the correlation between bacteria or fungi, reducing the relative abundance of Acetobacter and Paenibacillus in bacterial communities, and inhibiting the tendency of Monascus to replace Issatchenkia in fungal communities, thus delaying the aerobic spoilage process. Due to the prevention of the development of aerobic spoilage microorganisms, the silage injected with Lactobacillus buchneri or Bacillus licheniformis exhibited improved aerobic stability.

Lactational performance, ruminal fermentation, and enteric gas emission of dairy cows fed an amylase-enabled corn silage in diets with different starch concentrations.

This study investigated the effects of feeding an amylase-enabled corn silage (ACS) on the performance and enteric gas emissions in lactating dairy cows. Following a 2-wk covariate period, 48 mid-lactation Holstein cows were assigned to 1 of 3 treatments in a 10-wk randomized complete block design experiment. Treatments were diets containing the same proportion of corn silage (40% of dietary DM) as follows: (1) a conventional hybrid corn silage control (CON), (2) ACS replacing the control silage (ADR), and (3) the ADR diet replacing soybean hulls with ground corn grain to achieve the same dietary starch concentration as CON (ASR). Control corn silage and ACS were harvested on the same day and contained 40.3% and 37.1% DM and (% of DM): 37.2% and 41.0% NDF and 37.1% and 30.0% starch, respectively. Enteric gas emissions were measured using the GreenFeed system. Two cows were culled due to health-related issues during the covariate period. Ruminal fluid was collected from 24 cows (8 per treatment) using the orogastric ruminal sampling technique. When compared with CON, cows fed ADR had increased DMI during experimental wk 3, 4, and 9, but treatment did not affect milk or ECM milk yields (39.0 kg/d on average; SEM = 0.89). Compared with CON, feed efficiency (per unit of milk, but not ECM) tended to be lower for ADR, whereas milk true protein concentration (a tendency) and yield were lower for ASR. Milk urea N was decreased by both ADR and ASR diets relative to CON. Compared with CON, daily CH4 emission and emission intensity were increased by ADR but not ASR. Total protozoal count tended to be increased by both diets formulated with ACS when compared with control corn silage. Total-tract digestibility of dietary NDF was greater for ASR, and that of ADF was greater for both ADR and ASR versus CON. The molar proportion of acetate (a tendency) and acetate-to-propionate ratio were increased by ADR, but not ASR, when compared with CON. Replacement of CON with ACS (having lower starch concentration) in the diet of dairy cows increased DMI during the initial weeks of the experiment, maintained ECM, tended to decrease feed efficiency, and increased enteric CH4 emissions, likely due to increased intake of digestible fiber, compared with CON.

Effects of feeding different dietary rates of mixed fodder beet tops-wheat straw silage on the performance of Holstein lactating cows.

Rations containing different rates of the mixed fodder beet tops-wheat straw silage (BS), instead of corn silage (CS), were given to 30 mid-lactation Holstein cows (all in parity 2) to measure the effects on feed consumption, milk production efficiency, milk chemistry, urinary purine-derivatives (PD), blood chemistry, antioxidant levels, and in vitro methane (CH4) emission. The BS was prepared by mixing the fodder beet tops with wheat straw at a ratio of 9:1 based on fresh weight. The experimental design was completely randomized (one 28-d period with 21-d adaptation) using 30 cows (10 animals/treatment) and 3 treatments. The treatments were 1) a diet containing CS only (25 g/100 g DM) (CSD), 2) a diet containing 50% CS (12.5 g/100 g DM) and 50% BS (12.5 g/100 g DM) (CBSD), and 3) diet containing BS only (25 g/100 g DM) (BSD). Each animal (as an experimental unit) was housed individually in the tie stall and had ad libitum access to its diet. Dietary replacing 50% of CS with BS showed no significant differences in milk production, fat-corrected milk, fat and protein yields, feed efficiency, and apparent digestibility, however, these variables were less (P < 0.05) in the cows fed with BSD. Cows fed on BSD had less intakes of DM, organic matter, crude protein, and neutral detergent fiber but greater oxalic acid intake and blood urea-N, as compared to the other cows. Milk percentages of fat, protein, lactose, urea N, blood serum glucose, triglyceride, cholesterol, total protein, albumin, globulin, Ca, and P, as well as in vitro ruminal pH, were not affected by the diets. Saturated fatty acids concentration was less and monounsaturated FA and polyunsaturated FA (PUFA) was greater in the milk of cows receiving CBSD, compared to the other groups. The inclusion of both BS rates in the diet decreased the in vitro gas production, protozoa number, and CH4 emission in comparison to the control. Cows fed BSD had decreased levels of urinary allantoin, PD excreted or absorbed, and estimated microbial-N synthesis than the control and CBSD-fed groups. The milk and blood total antioxidant capacity (TAC) of the animals fed CBSD was the maximum among the cows. Overall, under the current experimental conditions, replacing 50% of dietary CS with BS did not affect milk production, but increased milk PUFA, as well as blood and milk TAC, and decreased in vitro CH4 emission, so it's feeding to lactating Holstein cows is recommended.

Effect of nicotinic acid supplementation on digestion, metabolism, microbiome, and production in late-lactation Holstein cows.

The purpose of this experiment was to determine if nicotinic acid (NA) effects on dairy cows and rumen microbial characteristics are forage type dependent (corn silage, CS; grass silage, GS). Four late lactation (days in milk = 225 +/- 12 d) Holstein cows were used in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. The main effects were a CS (66.10% CS) based diet or a GS (79.59%) based diet with or without 12 g/d NA. Each experimental period lasted for 28 d. Milk production and milk components, blood metabolites, apparent total-tract nutrient digestibilities, minutes rumen pH were below 5.8 as an indicator of ruminal acidosis, and body temperature changes were analyzed as indicators of heat stress. Nicotinic acid supplementation did not improve apparent total-tract nutrient digestibility. Feeding a GS-based diet improved NDF and hemicellulose digestibility. Feeding a CS-based diet increased the apparent total-tract digestibility of fat, and minutes rumen pH below 5.8 for a greater proportion of the time. The CS-based diet also improved milk yield, milk fat and protein yields, and energy-corrected milk yield; however, somatic cell count and BHB were also increased. Supplementing NA tended to decrease nonesterified fatty acids, especially when combined with GS where DMI was low. There was a trend for the total protozoa population to increase when GS and NA were fed but decreased when CS and NA were fed. Nicotinic acid tended to decrease rumen protozoal populations of Dasytricha, but increased populations of Ophryoscolex and Diplodiniinae with GS diets and decreased with CS diets. Entodiniinae were increased with CS but NA had no effect. Body temperature was increased when a CS-based diet was fed when compared with a GS-based diet. More research is needed to determine how NA can affect rumen microbial protein synthesis and what kind of diets will provide the optimum effect.

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.

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.

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.

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.

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.

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.

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.