Growth performance, blood metabolites, ruminal fermentation, and bacterial community in preweaning dairy calves fed corn silage-included starter and total mixed ration.

The objective of this study was to evaluate the effects of the inclusion of whole-plant corn silage (WPCS) in a starter or total mixed ration (TMR) on growth, blood metabolites, ruminal fermentation, and microbial community in preweaning dairy calves. A total of 45 healthy dairy calves were blocked by date of birth and randomly assigned to 1 of 3 treatments: 100% calf starter (CONS), a mix of 85% calf starter and 15% WPCS [dry matter (DM) basis; CSCS], or 100% WPCS-based lactation TMR (CTMR). Pasteurized normal milk was fed to all the animals under the same regimen. The experiment ran from when the calves were 2 d old to weaning at 63 d. Milk and feed intakes were recorded daily. Growth performance data and blood samples were collected on wk 3, 5, 7, and 9 of the experiment. Rumen fluid was sampled at 40 and 60 d. The 3 treatments had different particle size fractions. The CSCS group had greater medium fraction (<19 mm, >8 mm) and particles retained on 8-mm sieves than the other 2 groups, whereas the CTMR group had the greatest long (>19 mm) and fine (<4 mm) fractions and physically effective neutral detergent fiber (NDF) on 8- and 4-mm sieves, but had the smallest short fraction (<8 mm, >4 mm) and particles retained on 4-mm sieves. The 24-h in vitro digestibility of DM, crude protein (CP), NDF, and acid detergent fiber (ADF) were decreased in order by the CONS, CSCS, and CTMR groups. Compared with the CONS group, the digestibility of ether extract (EE) was lower in the CSCS and CTMR groups, whereas the digestibility of starch was similar among treatments. During the experimental period, the DM, CP, and metabolizable energy intakes from milk, solid feed, and total feed were not affected by treatments. The NDF, ADF, and EE intakes and potentially digestible intakes were greater in the CTMR group than in the other 2 groups. With the exception that body barrel was greater for calves fed CSCS, growth parameters and blood metabolites were similar among treatments. Compared with the CSCS group, the CTMR group had greater rumen pH and total volatile fatty acids, propionate, and isovalerate concentrations, but a lower acetate:propionate ratio. The CTMR group had greater relative abundances of some cellulolytic bacteria (Rikenellaceae RC9 gut group, Christensenellaceae R7, Ruminococcaceae NK4A214, Ruminococcaceae UCG, Ruminococcus, and Erysipelotrichaceae UCG) in the rumen, which may be beneficial for the early acquisition of specific adult-associated microorganisms. In summary, a WPCS-based lactation TMR, but not the WPCS-included starter, had the potential to be an alternative starter in preweaning calves without having significant adverse effects. These findings provide theoretical and practical implications for the rational application of TMR in the early life of dairy calves.

Competitive Analysis of Rumen and Hindgut Microbiota Composition and Fermentation Function in Diarrheic and Non-Diarrheic Postpartum Dairy Cows.

Postpartum dairy cows can develop nutritional diarrhea when their diet is abruptly changed for milk production. However, it is unclear whether nutritional diarrhea develops as a result of gut acidosis and/or dysbiosis. This study aimed to uncover changes in the gastrointestinal microbiota and its fermentation parameters in response to nutritional diarrhea in postpartum dairy cows. Rumen and fecal samples were collected from twenty-four postpartum cows fed with the same diet but with different fecal scores: the low-fecal-score (LFS: diarrheic) group and high-fecal-score (HFS: non-diarrheic) group. A microbiota difference was only observed for fecal microbiota, with the relative abundance of Defluviitaleaceae_UCG-011 and Lachnospiraceae_UCG-001 tending (p < 0.10) to be higher in HFS cows compared to LFS cows, and Frisingicoccus were only detected in HFS cows. The fecal bacterial community in LFS cows had higher robustness (p < 0.05) compared to that in HFS cows, and also had lower negative cohesion (less competitive behaviors) and higher positive cohesion (more cooperative behaviors) (p < 0.05) compared that in to HFS cows. Lower total volatile fatty acids and higher ammonia nitrogen (p < 0.05) were observed in LFS cows' feces compared to HFS cows. The observed shift in fecal bacterial composition, community networks, and metabolites suggests that hindgut dysbiosis could be related to nutritional diarrhea in postpartum cows.

Novel Process Methods for the Whole Cottonseed: Effect on the Digestibility, Productivity, Fat Profile, and Milk Gossypol Levels in Lactating Dairy Cows.

In this study, we aimed to determine the effect of mixed-process methods on the ruminal degradability of whole cottonseed (WCS) both in situ and in vitro, and the effect on the production performance of dairy cows. Eight WCS process methods were tested on the ruminal digestibility, including crush-alkali 1 (CA1), crush-alkali 2 (CA2), crush-alkali 3 (CA3), alkali 1-crush (A1C), alkali 2-crush (A2C), alkali 3-crush (A3C), crush-only (CO), and non-processed. Alkali 1, 2, and 3 indicate the supplementation of alkali to WCS at the dose of 4% on dry matter (DM) base as followed: 4% NaOH, 2% NaOH + 2% CaO, and 2% NaOH + 2% CaCl2 alkaline, respectively. Among all treatments, CA2 showed the highest WCS ruminal degradation in situ and the highest intestinal digestibility of WCS in vitro. Furthermore, an animal experiment was conducted for 60 days on 30 Holstein dairy cows, using a diet without WCS (CON group), a diet containing 8% non-processed WCS (NP group), and a diet containing 8% CA2-treated WCS (CA2 group). The results indicated that the dry matter intake, 4% fat-corrected milk production, milk protein, milk fat, and content of short-chain saturated fatty acid of milk in the CA2 group were significantly higher (P < 0.05) than CON group. Furthermore, DMI, the CLA was significantly greater (P < 0.05) in the CA2 group than the other groups. Additionally, the free gossypol concentration in serum or milk was under safety level in the three groups. Overall, crush and alkalization (NaOH: CaO = 1:1) treatment could improve the utilization of WCS in dairy farms.

Rumen Fermentation, Digestive Enzyme Activity, and Bacteria Composition between Pre-Weaning and Post-Weaning Dairy Calves.

To better understand the transition of rumen function during the weaning period in dairy calves, sixteen Holstein dairy calves were selected and divided into two groups: pre-weaning (age = 56 ± 7 day, n = 8) and post-weaning (age = 80 ± 6 day, n = 8). The rumen fluid was obtained by an oral gastric tube. The rumen fermentation profile, enzyme activity, bacteria composition, and their inter-relationship were investigated. The results indicated that the post-weaning calves had a higher rumen acetate, propionate, butyrate, and microbial crude protein (MCP) than the pre-weaning calves (p < 0.05). The rumen pH in the post-weaning calves was lower than the pre-weaning calves (p < 0.05). The protease, carboxymethyl cellulase, cellobiohydrolase, and glucosidase in the post-weaning calves had a lower trend than the pre-weaning calves (0.05 < p < 0.1). There was no difference in α and ß diversity between the two groups. Linear discriminant analysis showed that the phylum of Fibrobacteres in the post-weaning group was higher than the pre-weaning group. At the genus level, Shuttleworthia, Rikenellaceae, Fibrobacter, and Syntrophococcus could be worked as the unique bacteria in the post-weaning group. The rumen bacteria network node degree in the post-weaning group was higher than the pre-weaning group (16.54 vs. 9.5). The Shuttleworthia genus was highly positively correlated with MCP, propionate, total volatile fatty acid, glucosidase, acetate, and butyrate (r > 0.65, and p < 0.01). Our study provided new information about the rumen enzyme activity and its relationship with bacteria, which help us to better understand the effects of weaning on the rumen function.

Adsorbents Reduce Aflatoxin M1 Residue in Milk of Healthy Dairy Cow Exposed to Moderate Level Aflatoxin B1 in Diet and Its Exposure Risk for Humans.

This study investigated the effect of moderate risk level (8 µg/kg) AFB1 in diet supplemented with or without adsorbents on lactation performance, serum parameters, milk AFM1 content of healthy lactating cows and the AFM1 residue exposure risk in different human age groups. Forty late healthy lactating Holstein cows (270 ± 22 d in milk; daily milk yield 21 ± 3.1 kg/d) were randomly assigned to four treatments: control diet without AFB1 and adsorbents (CON), CON with 8 µg/kg AFB1 (dry matter basis, AF), AF + 15 g/d adsorbent 1 (AD1), AF + 15 g/d adsorbent 2 (AD2). The experiment lasted for 19 days, including an AFB1-challenge phase (day 1 to 14) and an AFB1-withdraw phase (day 15 to 19). Results showed that both AFB1 and adsorbents treatments had no significant effects on the DMI, milk yield, 3.5% FCM yield, milk components and serum parameters. Compared with the AF, AD1 and AD2 had significantly lower milk AFM1 concentrations (93 ng/L vs. 46 ng/L vs. 51 ng/L) and transfer rates of dietary AFB1 into milk AFM1 (1.16% vs. 0.57% vs. 0.63%) (p < 0.05). Children aged 2-4 years old had the highest exposure risk to AFM1 in milk in AF, with an EDI of 1.02 ng/kg bw/day and a HI of 5.11 (HI > 1 indicates a potential risk for liver cancer). Both AD1 and AD2 had obviously reductions in EDI and HI for all population groups, whereas, the EDI (≥0.25 ng/kg bw/day) and HI (≥1.23) of children aged 2-11 years old were still higher than the suggested tolerable daily intake (TDI) of 0.20 ng/kg bw/day and 1.00 (HI). In conclusion, moderate risk level AFB1 in the diet of healthy lactating cows could cause a public health hazard and adding adsorbents in the dairy diet is an effective measure to remit AFM1 residue in milk and its exposure risk for humans.

Effects of Age, Diet CP, NDF, EE, and Starch on the Rumen Bacteria Community and Function in Dairy Cattle.

To understand the effects of diet and age on the rumen bacterial community and function, forty-eight dairy cattle at 1.5 (M1.5), 6 (M6), 9 (M9), 18 (M18), 23 (M23), and 27 (M27) months old were selected. Rumen fermentation profile, enzyme activity, and bacteria community in rumen fluid were measured. The acetate to propionate ratio (A/P) at M9, M18, and M23 was higher than other ages, and M6 was the lowest (p < 0.05). The total volatile fatty acid (TVFA) at M23 and M27 was higher than at other ages (p < 0.05). The urease at M18 was lower than at M1.5, M6, and M9, and the xylanase at M18 was higher than at M1.5, M23, and M27 (p < 0.05). Thirty-three bacteria were identified as biomarkers of the different groups based on the linear discriminant analysis (LDA) when the LDA score >4. The variation partitioning approach analysis showed that the age and diet had a 7.98 and 32.49% contribution to the rumen bacteria community variation, respectively. The richness of Succinivibrionaceae_UCG-002 and Fibrobacter were positive correlated with age (r > 0.60, p < 0.01) and positively correlated with TVFA and acetate (r > 0.50, p < 0.01). The Lachnospiraceae_AC2044_group, Pseudobutyrivibrio, and Saccharofermentans has a positive correlation (r > 0.80, p < 0.05) with diet fiber and a negative correlation (r < -0.80, p < 0.05) with diet protein and starch, which were also positively correlated with the acetate and A/P (r > 0.50, p < 0.01). The genera of Lachnospiraceae_AC2044_group, Pseudobutyrivibrio, and Saccharofermentans could be worked as the target bacteria to modulate the rumen fermentation by diet; meanwhile, the high age correlated bacteria such as Succinivibrionaceae_UCG-002 and Fibrobacter also should be considered when shaping the rumen function.

Effects of Rumen-Protected Niacin on Dry Matter Intake, Milk Production, Apparent Total Tract Digestibility, and Faecal Bacterial Community in Multiparous Holstein Dairy Cow during the Postpartum Period.

Extensive studies about rumen-protected niacin (RPN) supplementation on dairy cows in early-lactation have been done, but the effects of RPN on changes in dry matter intake (DMI), milk production, feed digestibility, and fecal bacterial community were conflicting. The aim of this study was to investigate them affected by RPN in postpartum cows. Multiparous Holstein dairy cows (n = 12, parity = 3.5 ± 0.5, body weights = 740 ± 28 kg) were divided into two groups supplemented with either 0 (CON) or 20 g/d RPN (RPN). Our results showed that RPN supplementation increased DMI and milk production of cows during the first three weeks after calving (p < 0.05). The concentrations of neuropeptide Y and orexin A were significantly higher in RPN group than that in the CON group during postpartum period (p < 0.05). The apparent total-tract digestibility of nutrients was similar between the CON and RPN groups at 2 weeks after calving (p > 0.05). The 16S rRNA gene sequencing analysis showed that RPN had no impact on the alpha and beta diversity, although 4 genera were changed in cow feces at 14 days after calving. Overall, 20 g/d RPN added to the diet could improve DMI and milk yield up to two weeks after calving with little influence on feed digestibility.

Effects of Different Parts on the Chemical Composition, Silage Fermentation Profile, In Vitro and In Situ Digestibility of Paper Mulberry.

Paper mulberry (Broussonetia papyrifera, PM) is high protein but unutilized as a feed source. The study explores the different parts (leaf, stem, and whole plant) of PM chemical composition, silage fermentation, and in vitro and in situ digestibility, aiming to give some guidelines to PM usage as feed. The result showed that the leaf had a higher fresh weight than the stem (p < 0.05). The dry matter contents of the three groups had no differences. The highest crude protein, ether extract, water-soluble carbohydrate, ash, calcium, phosphorus, amino acid contents, and butter capacity were observed in the leaf (p < 0.05). The stem had the highest (p < 0.05) neutral detergent fiber, acid detergent fiber, and lignin contents. After ensiling, the stem silage had the lowest pH value, ammonia nitrate (NH3-N), lactate, acetate, and propionate (p < 0.05). The leaf silage had the highest pH value (p < 0.05). The lactate, acetate, and propionate in the leaf and whole plant silage had no difference. The butyrate was not detected in all silage. The in vitro and in situ digestibility experiments showed the leaf had the highest digestibility (p < 0.05), which could produce more volatile fatty acids and have a higher effective digestibility. These results allow a greater understanding of PM to be used as a feedstuff.

Effects of Paper Mulberry Silage on the Milk Production, Apparent Digestibility, Antioxidant Capacity, and Fecal Bacteria Composition in Holstein Dairy Cows.

Paper mulberry (Broussonetia papyrifera; PM) is an excellent and extensive type of roughage in Asia. This study aimed to evaluate the effects of PM silage on the milk production, apparent digestibility, antioxidant capacity, and fecal bacteria composition in Holstein dairy cows. Forty-five lactating Holstein dairy cows with a similar milk yield and parity were selected and randomly assigned to three groups. The control group was fed a non-PM silage diet, and the PM-treated groups were fed 4.5 and 9.0% PM silage supplementary diets for 28 days. Then, treatment groups were fed diets containing 13.5 and 18.0% PM silage for the next 28 days, respectively. PM silage increased the milk urea nitrogen and decreased the somatic cell count (p < 0.05), but did not affect the dry matter intake, milk yield, apparent digestibility, and energy balance of dairy cows. PM silage can enhance the blood total antioxidant capacity, superoxide dismutase, and immune globulin content (p < 0.05). The PM silage significantly decreased the relative abundance of the genera Ruminococcaceae UCG-013 and Tyzzerella-4 (p < 0.05). In conclusion, PM silage enhanced the antioxidant capacity and immunity of dairy cows, but did not influence the milk yield, dry matter digestibility, and fecal bacteria composition.

The day-to-day stability of the ruminal and fecal microbiota in lactating dairy cows.

In this study, we examined differences between the microbiota of the ruminal fluid (DR) and feces (DF) from five lactating dairy cows over three consecutive days using 16S rRNA gene sequence-based analysis. Results showed significant differences between the microbial communities of the DR and DF. In particular, the relative abundance of the phyla Firmicutes and Actinobacteria was significantly lower (q < 0.001) in DR compared with DF, while the relative abundance of Bacteroidetes was significantly higher in DF than that of DR (q < 0.001). A significantly higher relative abundance of the genera Bifidobacterium, 5-7N15, Clostridium, Epulopiscium, SMB53, Turicibacter, Dorea, Roseburia, and Akkermansia was observed in the DF, while a higher relative abundance of the genera Prevotella, Butyrivibrio, CF231, RFN20, and Succiniclasticum was observed in the DR. A further analysis using the functional prediction program PICRUSt showed that sequences belonging to the 5-7N15, Akkermansia, Bifidobacterium, Clostridium, Dorea, Epulopiscium, Roseburia, and Turicibacter were significantly and positively correlated with glycan biosynthesis and metabolism, while CF231, Prevotella, RFN20, and Succiniclasticum were significantly and positively correlated with amino acid, lipid, carbohydrate, other amino acid, cofactors, and vitamins metabolism. No significant differences were observed across the three consecutive days in either the DR or DF ecosystems, with no significant differences in the diversity or abundance at the phylum and genus levels suggested that there is a limited day-to-day variability in the gut microbiota.

Effects of sulfanilamide on boar sperm quality, bacterial composition, and fertility during liquid storage at 17°C.

Sulfanilamide (SA) is an effective broad-spectrum antibacterial agent in human and veterinary medicine. The purpose of this study was to evaluate the effects of SA on boar sperm quality during liquid storage at 17°C and determine the optimal concentration of SA and its effects on bacterial growth, microbial composition, and maternal fertility. Boar ejaculates were diluted with a basic extender, containing different concentrations of SA, and stored in a 17°C incubator for 6 days. The sperm motility, plasma membrane integrity, and acrosome integrity were measured daily. The results showed that when the concentration of SA was 0.02 g/L, the sperm quality parameters were significantly higher than those of all other treatment groups (p < .05). We also monitored the bacterial growth and compared the differences in the microbial species between the 0.02 g/L SA group and the control by 16S rDNA sequencing. The results revealed that some bacteria, such as Staphylococcus and Pseudomonas, were considerably lower in the 0.02 g/L SA group than in the control group (p < .05). In addition, preserved semen was used for artificial insemination, and results showed that 0.02 g/L SA group had a higher litter size, and its pregnancy rate was 92.5%.

The effects of different levels of superoxide dismutase in Modena on boar semen quality during liquid preservation at 17°C.

This study was conducted to investigate the influence of superoxide dismutase (SOD) on the quality of boar semen during liquid preservation at 17°C. Semen samples from 10 Duroc boars were collected and pooled, divided into five equal parts and diluted with Modena containing different concentrations (0, 100, 200, 300 and 400 U/mL) of SOD. During the process of liquid preservation at 17°C, sperm motility, acrosome integrity, membrane integrity, total antioxidant capacity (T-AOC) activity, malondialdehyde (MDA) content and hydrogen peroxide (H2 O2 ) content were measured and analyzed every 24 h. Meanwhile, effective survival time of boar semen during preservation was evaluated and analyzed. The results indicated that different concentrations of SOD in Modena showed different protective effects on boar sperm quality. Modena supplemented with SOD decreased the effects on reactive oxygen species on boar sperm quality during liquid preservation compared with that of the control group. The added 200 U/mL SOD group showed higher sperm motility, membrane integrity, acrosome integrity, effective survival time and T-AOC activity. Meanwhile, the added 200 U/mL SOD group showed lower MDA content and H2 O2 content. In conclusion, addition of SOD to Modena improved the boar sperm quality by reducing oxidative stress during liquid preservation at 17°C and the optimum concentration was 200 U/mL.