Partially substituting alfalfa hay with hemp forage in the diet of goats improved feed efficiency, ruminal fermentation pattern and microbial profiles.

The use of hemp as a forage source in livestock diets has been less studied because bioactive residues in animal tissues may pose a risk to consumers. This study investigated the effects of partial substitution of alfalfa hay (AH) with hemp forage (HF) in growing goat diets on growth performance, carcass traits, ruminal fermentation characteristics, rumen microbial communities, blood biochemistry, and antioxidant indices. Forty Xiangdong black goats with body weight (BW) 7.82 ± 0.57 kg (mean ± SD) were grouped by BW and randomly assigned into one of the four treatment diets (n = 10/treatment) in a completely randomized design. The goats were fed ad libitum total mixed rations containing 60% forage and 40% concentrate (DM basis). The diets included control (CON; 60% AH and 40% concentrate), 55% AH and 5% HF (HF5), 50% AH and 10% HF (HF10), and 40% AH and 20% HF (HF20). Increasing the substitution of HF for AH linearly decreased (P < 0.01) DM intake and improved feed conversion efficiency. However, final BW, average daily gain, carcass traits, meat quality, and most blood biochemistry indices did not differ among treatments. The ruminal NH3-N concentration and blood urine nitrogen linearly increased (P < 0.01) with increasing substitution rate of HF, whereas the total volatile fatty acids concentration quadratically changed (P < 0.01). Substitution of AH with HF had no effect on the diversity and richness of ruminal microbes, though it linearly decreased (P = 0.040) Prevotella_1 and linearly increased (P = 0.017) Rikenellaceae_RC9_gut_group. The cannabinoids and/or their metabolites were detected in both ruminal filtrates (8) and plasma (4), however, no detectable cannabinoid-related residues were observed in meat. These results indicate that the HF could be used to partially substitute AH in goat diets, whereas the effects vary between substitution rates of HF for AH. Although no cannabinoid-related residues were detected in meat, the presence of cannabinoids residues in blood warrants further study of HF feeding to confirm the cannabinoids residues are not present in the animal products.

Rolapitant treats lung cancer by targeting deubiquitinase OTUD3.

BACKGROUND: Lung cancer is cancer with the highest morbidity and mortality in the world and poses a serious threat to human health. Therefore, discovering new treatments is urgently needed to improve lung cancer prognosis. Small molecule inhibitors targeting the ubiquitin-proteasome system have achieved great success, in which deubiquitinase inhibitors have broad clinical applications. The deubiquitylase OTUD3 was reported to promote lung tumorigenesis by stabilizing oncoprotein GRP78, implying that inhibition of OTUD3 may be a therapeutic strategy for lung cancer. RESULTS: In this study, we identified a small molecule inhibitor of OTUD3, Rolapitant, by computer-aided virtual screening and biological experimental verification from FDA-approved drugs library. Rolapitant inhibited the proliferation of lung cancer cells by inhibiting deubiquitinating activity of OTUD3. Quantitative proteomic profiling indicated that Rolapitant significantly upregulated the expression of death receptor 5 (DR5). Rolapitant also promoted lung cancer cell apoptosis through upregulating cell surface expression of DR5 and enhanced TRAIL-induced apoptosis. Mechanistically, Rolapitant directly targeted the OTUD3-GRP78 axis to trigger endoplasmic reticulum (ER) stress-C/EBP homologous protein (CHOP)-DR5 signaling, sensitizing lung cancer cells to TRAIL-induced apoptosis. In the vivo assays, Rolapitant suppressed the growth of lung cancer xenografts in immunocompromised mice at suitable dosages without apparent toxicity. CONCLUSION: In summary, the present study identifies Rolapitant as a novel inhibitor of deubiquitinase OTUD3 and establishes that the OTUD3-GRP78 axis is a potential therapeutic target for lung cancer.

A novel nucleic acid linker for multi-gene expression enhances plant and animal synthetic biology.

The production of compact vectors for gene stacking is hindered by a lack of effective linkers. Here, we report that a 26-nt nucleic acid linker, NAL1, from the fungus Glarea lozoyensis and its truncated derivatives could connect two genes as a bicistron, enabling independent translation in a maize protoplast transient expression system and human 293 T cells. The optimized 9-nt NAL10 linker was then used to connect four genes driven by a bidirectional promoter; this combination was successfully used to reconstruct the astaxanthin biosynthesis pathway in transgenic maize. The short and efficient nucleic acid linker NAL10 can be widely used in multi-gene expression and synthetic biology in animals and plants.

Identification and characterization of yellow stripe-like genes in maize suggest their roles in the uptake and transport of zinc and iron.

BACKGROUND: Yellow Stripe-Like (YSL) proteins are involved in the uptake and transport of metal ions. They play important roles in maintaining the zinc and iron homeostasis in Arabidopsis, rice (Oryza sativa), and barley (Hordeum vulgare). However, proteins in this family have not been fully identified and comprehensively analyzed in maize (Zea mays L.). RESULTS: In this study, we identified 19 ZmYSLs in the maize genome and analyzed their structural features. The results of a phylogenetic analysis showed that ZmYSLs are homologous to YSLs of Arabidopsis and rice, and these proteins are divided into four independent branches. Although their exons and introns have structural differences, the motif structure is relatively conserved. Analysis of the cis-regulatory elements in the promoters indicated that ZmYSLs might play a role in response to hypoxia and light. The results of RNA sequencing and quantitative real-time PCR analysis revealed that ZmYSLs are expressed in various tissues and respond differently to zinc and iron deficiency. The subcellular localization of ZmYSLs in the protoplast of maize mesophyll cells showed that they may function in the membrane system. CONCLUSIONS: This study provided important information for the further functional analysis of ZmYSL, especially in the spatio-temporal expression and adaptation to nutrient deficiency stress. Our findings provided important genes resources for the maize biofortification.

Effects of dietary Clostridium butyricum and rumen protected fat on meat quality, oxidative stability, and chemical composition of finishing goats.

BACKGROUND: Clostridium butyricum (CB) is a probiotic that can regulate intestinal microbial composition and improve meat quality. Rumen protected fat (RPF) has been shown to increase the dietary energy density and provide essential fatty acids. However, it is still unknown whether dietary supplementation with CB and RPF exerts beneficial effects on growth performance and nutritional value of goat meat. This study aimed to investigate the effects of dietary CB and RPF supplementation on growth performance, meat quality, oxidative stability, and meat nutritional value of finishing goats. Thirty-two goats (initial body weight, 20.5 ± 0.82 kg) were used in a completely randomized block design with a 2 RPF supplementation (0 vs. 30 g/d) × 2 CB supplementation (0 vs. 1.0 g/d) factorial treatment arrangement. The experiment included a 14-d adaptation and 70-d data and sample collection period. The goats were fed a diet consisted of 400 g/kg peanut seedling and 600 g/kg corn-based concentrate (dry matter basis). RESULT: Interaction between CB and RPF was rarely observed on the variables measured, except that shear force was reduced (P < 0.05) by adding CB or RPF alone or their combination; the increased intramuscular fat (IMF) content with adding RPF was more pronounced (P < 0.05) with CB than without CB addition. The pH24h (P = 0.009), a* values (P = 0.007), total antioxidant capacity (P = 0.050), glutathione peroxidase activities (P = 0.006), concentrations of 18:3 (P < 0.001), 20:5 (P = 0.003) and total polyunsaturated fatty acids (P = 0.048) were increased, whereas the L* values (P < 0.001), shear force (P = 0.050) and malondialdehyde content (P = 0.044) were decreased by adding CB. Furthermore, CB supplementation increased essential amino acid (P = 0.027), flavor amino acid (P = 0.010) and total amino acid contents (P = 0.024) as well as upregulated the expression of lipoprotein lipase (P = 0.034) and peroxisome proliferator-activated receptor γ (PPARγ) (P = 0.012), and downregulated the expression of stearoyl-CoA desaturase (SCD) (P = 0.034). The RPF supplementation increased dry matter intake (P = 0.005), averaged daily gain (trend, P = 0.058), hot carcass weight (P = 0.046), backfat thickness (P = 0.006), concentrations of 16:0 (P < 0.001) and c9-18:1 (P = 0.002), and decreased the shear force (P < 0.001), isoleucine (P = 0.049) and lysine content (P = 0.003) of meat. In addition, the expressions of acetyl-CoA carboxylase (P = 0.003), fatty acid synthase (P = 0.038), SCD (P < 0.001) and PPARγ (P = 0.022) were upregulated due to RPF supplementation, resulting in higher (P < 0.001) content of IMF. CONCLUSIONS: CB and RPF could be fed to goats for improving the growth performance, carcass traits and meat quality, and promote fat deposition by upregulating the expression of lipogenic genes of Longissimus thoracis muscle.

Effects of castration and eucalyptus oil supplementation on growth performance, nutrient digestibility, and blood-immunity indicators of male Holstein calves.

The purpose of this study was to investigate the effects of early castration and eucalyptus oil (EUC) supplementation on dry matter intake (DMI), growth performance, and immune response of Holstein calves. Fifty-six male Holstein calves 52 d old and with an initial body weight (BW) of 63.5 ± 5.27 kg were used. The animals were blocked by BW and randomly assigned into 1 of the 4 treatment groups in a randomized complete block design with a 2 (no castration vs. castration) × 2 (without vs. with EUC) factorial arrangement of treatments. The treatments were (1) uncastrated calves fed without EUC, (2) uncastrated calves fed 0.5 g/d EUC (EUC group), (3) castrated calves (steers) fed without EUC (castrated group), and (4) steers fed with 0.5 g/d EUC (castrated + EUC). The experiment was 8 wk long, including pre- and postweaning (weaned at 72 d). The EUC × castrated interactions were not significant for DMI, growth performance, nutrient digestibility, and immune response. Castration did not affect the DMI, final BW, average daily gain (ADG), or feed efficiency, except that the ADG was greater for bull calves than for steers at postweaning. Supplementation with EUC increased DMI pre- and postweaning and increased the ADG of weaned calves. Digestibility in the total digestive tract was not affected by castration (except for organic matter digestibility), whereas adding EUC improved the digestibility of dry matter, acid detergent fiber, and crude protein. Blood concentration of IL-6 at d 94 was decreased by feeding EUC. These results indicate that the EUC could be fed to either intact or castrated dairy calves to promote growth and health postweaning; castration before weaning may reduce ADG and cause inflammatory stress without affecting feed intake or feed efficiency.

ZmGluTR1 is involved in chlorophyll biosynthesis and is essential for maize development.

Chlorophyll is the most important carrier of photosynthesis in plants and is therefore vital for plant growth and development. Synthesis of 5-aminolevulinic acid (ALA) is initiated and catalyzed by glutamyl-tRNA reductase (GluTR) and is the rate-limiting step in chlorophyll biosynthesis. GluTR is controlled by several regulating factors. Although many studies have investigated the structure and function of GluTR in plants, the maize (Zea mays L.) GluTR has not yet been reported. Here, we isolated and identified the first loss-of-function mutant of GluTR in plants from a maize mutagenic population. The stop-gain mutation in ZmGluTR1 resulted in leaf etiolation throughout the growing season. The level of intermediates of chlorophyll biosynthesis and photosynthetic pigments decreased markedly and abnormal chloroplast structure was also observed in the mutants. Further analysis revealed that the deletion of carboxyl terminal (C-terminal) led to premature transcription termination and this hindered the interaction with FLUORESCENT (FLU), thereby influencing the stability of mutated ZmGluTR1 and leading to abolish interaction with GluTR-binding protein (GluBP). Moreover, mutations in the catalytic domain or nicotinamide adenine dinucleotide phosphate (NADPH) binding domain were lethal under normal growth conditions. These results indicate that ZmGluTR1 plays a fundamental role in chlorophyll biosynthesis and maize development.

Selective Passivation of Surface toward Bright Yellow Defective Emission of CdS Quantum Dots.

CdE (E = S, Se) quantum dots (QDs) with a broad and large Stokes shift PL emission have emerged as potential materials for white-light LEDs. However, this surface-related emission of nanocrystals is currently limited by low quantum efficiency. Herein, a convenient noninjected one-pot method at a relatively low temperature to prepare CdS QDs was readily achieved. The CdS-368 QD displays intense broad yellow emission in both solution and the solid state at room temperature. The coligation of organic and inorganic ligands passivates the electron trap states at the QD surface and suppresses nonradiative recombination, which is responsible for the high stability of colloids in organic solvents and the distinct fluorescence quantum yield.

Doping transition metals to modulate the chirality and photocatalytic activity of rare earth clusters.

In this paper, we report the successful assembly of achiral {Ln6M} ([Ln6M(µ3-OH)8(acac)12(CH3O)x(CH3OH)y], Ln = La, M = Mn, Co, Fe) and chiral {Nd9Fe2} ([Nd9Fe2(µ4-O)(µ3-OH)14(acac)16(NO3)(CH3OH)2(H2O)3]) rare earth clusters using achiral rigid ligands and a transition metal doping strategy. {Ln6M} can be viewed as the fusion of two {Ln3M} tetrahedrons by sharing vertices. {Nd9Fe2} results from the fusion of four {Ln3M} tetrahedrons by vertice and edge sharing. The substitution of Ln with transition metal leads to changes in the coordination pattern around neighboring Ln, which triggers the switch of metal center chirality. This study demonstrates the potentiality of utilizing transition metal doping and rigid ligand to control the chirality of rare earth clusters. In addition, the photocatalytic CO2 activity of these transition metal-doped rare earth clusters has been studied.

Effect of forage types differing in undigested neutral detergent fiber concentration and forage inclusion rate on reticulo-ruminal motility and fermentation, total tract barrier function, and blood metabolites of finishing beef heifers.

This study evaluated the effects of forages (BarS vs. STR) that differ in the uNDF concentration and FI rate on ruminal fermentation, total tract barrier function, reticulo-ruminal motility, and blood metabolites of beef heifers. Six ruminally cannulated Hereford × Simmental heifers (699 ± 69.1 kg BW) were used in a 6 × 6 Latin square (26 d periods) with a 2 × 3 factorial treatment arrangement. However, 1 heifer was removed from the study after period 2 due to health problems unrelated to treatment, resulting in an incomplete 6 × 6 Latin square design. Barley grain-based diets were formulated using BarS or wheat STR to alter uNDF (7.1% vs. 8.5% DM) with FI rates of 5%, 10%, or 15% of DM. There were limited interactions between the forage type and FI. DM intake was not affected (P ≥ 0.10) by forage type or FI. Use of STR vs. BarS increased uNDF intake (P < 0.001). Increasing FI increased (P < 0.001) uNDF intake for those fed 15% forage. Ruminal pH was not affected (P ≥ 0.10) by forage type; however, cattle fed 5% FI had lesser (P = 0.017) mean ruminal pH and maximum pH (P = 0.018) than those fed 10% and 15% of forage. The total SCFA concentration was not affected by forage type (P = 0.84) but cattle fed the 5% FI rate had lesser (P < 0.001) molar proportion of acetate when compared with cattle fed 10% and 15% forage. Increasing the FI rate decreased the molar proportion of propionate (P < 0.001). Feeding STR relative to BarS decreased (P = 0.041) the reticulo-ruminal contraction duration. In contrast, cattle fed the 10% and 15% FI rates had a greater (P = 0.028) contraction frequency with lower (P = 0.048) contraction area than those fed 5% forage. Plasma glucose, serum insulin, and serum amyloid A were not affected by forage type or FI rate (P ≥ 0.10). Cattle fed 15% forage had lesser (P = 0.040) concentration of serum haptoglobin when compared with cattle fed 5% or 10% forage. In conclusion, forage type used to affect the dietary uNDF concentration, and FI rate act independently suggesting that the provision of STR to increase uNDF reduces reticulo-ruminal contraction duration and total tract permeability but may not affect ruminal pH. Increasing the FI increased dietary uNDF, stabilized ruminal pH, stimulated more frequent reticulo-ruminal contractions, and may decrease the permeability of the gastrointestinal tract and systemic inflammation.

As a strategy to improve performance, feedlot cattle are often fed diets containing a high proportion of grain with minimal forage. However, diets with insufficient fiber may predispose cattle to nutritional disorders, such as ruminal acidosis, which leads to poor performance and increases production costs. Given the growing concern regarding minimal fiber requirement, the present study was designed to investigate the effects of forages differing in undigested neutral detergent fiber (uNDF; barley silage [BarS] vs. straw [STR]) concentration and forage inclusion (FI) rate (5%, 10%, or 15% of dietary dry matter [DM]) on reticulo-ruminal motility and fermentation, total tract barrier function, and blood metabolites of beef cattle. The inclusion of STR increased uNDF intake and rumination rate, but decreased reticulo-ruminal contraction duration, with no effect on ruminal pH and total short-chain fatty acid (SCFA) concentration. Decreasing the FI rate decreased uNDF intake, ruminating time, ruminal pH, and reticular contraction frequency, but increased the molar proportion of propionate and concentration of serum haptoglobin. In conclusion, forage type and FI rate act independently suggesting that providing forages that increase dietary uNDF may stimulate the frequency of reticulo-ruminal contractions without affecting ruminal pH. However, increasing FI inadvertently increased dietary uNDF, stabilized ruminal pH, increased rumination time, stimulated ruminal contractions, and decreased indicators of systemic inflammation.

Effect of physically effective neutral detergent fiber and undigested neutral detergent fiber on eating behavior, ruminal fermentation and motility, barrier function, blood metabolites, and total tract digestibility in finishing cattle.

This study evaluated the effects of physically effective neutral detergent fiber (peNDF) and undigested neutral detergent fiber (uNDF) on eating behavior, ruminal fermentation and motility, barrier function, blood metabolites, and total tract nutrient digestibility for finishing cattle. Six Simmental heifers (668 ± 28.4 kg BW) were used in a replicated 3 × 3 Latin square (21 d periods) balanced for carry-over effects. Treatments included a control (CON; Table 1) with no forage peNDF and minimal uNDF (peNDF: 0.0%, and uNDF: 4.88 ± 0.01; 95.15% barley grain, 4.51% vitamin and mineral supplement, and 0.34% urea on a DM basis). Pelleted wheat straw (PELL) was included at 10% of dietary DM by replacing barley grain to provide added uNDF but no forage peNDF (peNDF: 0.00%, and uNDF: 6.78 ± 0.02%). Finally, chopped wheat straw (STR) was included as a replacement for pelleted wheat straw to provide forage peNDF and uNDF (peNDF: 1.74 ± 0.06%, and uNDF: 6.86 ± 0.03%). Dry matter intake was not affected (P = 0.93) by treatments. Cattle fed CON spent less time ruminating (P = 0.010) and had less meals/d (P = 0.035) when compared with cattle fed STR, with those fed PELL being intermediate but not different from other treatments. Cattle fed CON had lesser ruminal pH (P = 0.020), and a greater duration that pH was < 5.5 (P = 0.020) as compared to cattle fed STR, with those fed PELL being intermediate but not different. Cattle fed CON and PELL had greater total short-chain fatty acid concentration (P = 0.003) and molar proportion of propionate (P < 0.001) when compared with cattle fed STR. Cattle fed STR had greater (P = 0.010) total ruminal pool size when compared with cattle fed CON and PELL. Cattle fed CON had greater (P = 0.043) duration between ruminal contractions when compared with cattle fed STR, with those fed PELL being intermediate. Cattle fed CON had greater serum amyloid A (P = 0.003) and haptoglobin (P < 0.001) concentration when compared with the other treatments. Cattle fed CON had greater dry matter (P < 0.001) digestibility when compared with the other treatments. In conclusion, inclusion of PELL and STR impacted eating behavior, but only STR affected ruminal fermentation, ruminal motility, systemic inflammation, and total tract nutrient digestibility in finishing cattle. These results are interpreted to suggest that the combination of peNDF and uNDF may be better than uNDF alone to adequately capture biological effects of NDF in high-concentrate diets.

Normally forages are included at minimal levels when formulating finishing cattle diets due to lower digestibility and higher operational and economic costs than concentrates. However, insufficient fiber may increase the risk of nutritional disorders such as ruminal acidosis, negatively impacting health, and consequently growth performance of feedlot cattle. An understanding of the minimum forage requirement and the constituent factors that best explain that requirement can help to promote health and performance while minimizing cost. The present study compared the effects of physically effective neutral detergent fiber (peNDF) and undigested neutral detergent fiber (uNDF) in concentrate-based finishing diets. Feeding more peNDF increased ruminating time and ruminal pH, but decreased time between ruminal contractions, total short-chain fatty acid concentration, and the molar proportion of propionate when compared with feeding a diet with minimal peNDF and uNDF. Responses to feeding uNDF with no peNDF were generally intermediate but not different from other treatments. In addition, feeding more peNDF decreased indicators of systemic inflammation and dry matter digestibility when compared with the diet with less peNDF or uNDF. In conclusion, peNDF, or at least the combination of peNDF and uNDF may be better than uNDF alone to stimulate rumination and ruminal motility, thereby stabilizing ruminal pH. Using both peNDF and uNDF may be useful in characterizing the value of fiber in finishing diets fed to feedlot cattle.

The ileal microbiome and mucosal immune profiles in response to dietary supplementation of ultra-grinded Astragalus membranaceus in weaned goats.

Weaning goats are susceptible to diarrhea and have weakened immune functions due to physiological, dietary and environmental stresses. Astragalus membranaceus (A. membranaceus), a traditional Chinese medicinal herb, has been shown to improve growth performance and immunity in weaned ruminants. However, the influence mechanism of A. membranaceus on intestinal microbiota and mucosal immunity in weaned goats is still unknown. This study investigated the effects of ultra-grinded A. membranaceus (UGAM) on the immune function and microbial community in the ileum of weaned goats. Eighteen healthy weaned Xiangdong black goats (BW, 5.30 ± 1.388 kg) were used in a study of completely randomized block design with 28 days long. The animals were randomly assigned to either a basal diet supplemented with 10 g/d of milk replacer (CON, n = 9) or the CON diet supplemented with 10 g/head UGAM (UGAM, n = 9). Supplementation of UGAM increased (p < 0.05) the plasma concentrations of total protein and albumin. Meanwhile, the addition of UGAM reduced (p < 0.05) the relative mRNA expression of the IL-6 gene (a marker of inflammation), indicating the potential immunomodulatory effect of UGAM. Moreover, the relative abundances of Verrucomicrobiota and Mycoplasma were lower (p < 0.05) in the ileum of goats supplemented with UGAM than CON. These findings suggest that dietary supplementation of UGAM may have enhanced the ileum health of weaned goats by reducing inflammation factor expression and reducing the relative abundance of pathogenic microbes. The observed beneficial effects of ultra-grinded A. membranaceus on ileal mucosal immune and the community of ileal microbiota indicate its potential to be used as a viable option for promoting the well-being of weaned goats under weaning stress.

The gas production, ruminal fermentation parameters, and microbiota in response to Clostridium butyricum supplementation on in vitro varying with media pH levels.

The aim of this study was to investigate the gas production (GP), dry matter disappearance (DMD), fermentation parameters, and rumen microbiota in response to Clostridium butyricum (CB) supplementation in batch culture using a high forage substrate. The doses of CB were supplemented at 0 (Control), 0.5 × 106, 1 × 106, and 2 × 106 CFU/bottle, respectively, at either media pH 6.0 or pH 6.6. The 16S rRNA gene sequencing was used to detect the microbiota of fermentation culture in control and 1 × 106 CFU/bottle after 24 h of incubation. The results showed that the GP (p < 0.001), DMD (p = 0.008), total volatile fatty acid (VFA) concentration (p < 0.001), acetate to propionate ratio (p < 0.001), and NH3-N concentration (p < 0.001) were greater at media pH 6.6 than pH 6.0. Furthermore, the linearly increased DMD (pH 6.0, p = 0.002; pH 6.6, p < 0.001) and quadratically increased butyrate proportion (pH 6.0, p = 0.076; pH 6.6, p < 0.053) and NH3-N concentration (pH 6.0, p = 0.003; pH 6.6, p = 0.014) were observed with increasing doses of CB. The Alpha diversity indexes of OTU number and Chao1 were higher (p = 0.045) at media pH 6.6 than pH 6.0, but they were not affected by CB supplementation. The PCoA analysis (unweighted uniFrac) demonstrated that the clustering of the bacterial microbiota of control and CB were distinctly separated from each other at media pH 6.0. At the phylum level, the abundance of Bacteroidota (p < 0.001) decreased, whereas that of Firmicutes (p = 0.026) increased when the media pH was elevated from 6.0 to 6.6. Supplementation of CB increased relative abundances of Rikenellaceae_RC9_gut_group (p = 0.002), Christensenellaceae_R-7_group (p < 0.001), and NK4A214_group (p = 0.002) at genus level. Interactions between media pH and CB addition were observed for bacteria at both phylum and genus levels. These results indicated that increasing the media pH level and CB supplementation increased in vitro rumen digestibility, and altered the ruminal fermentation pattern (by media pH) and microbiota.

Comparison of rumen bacterial communities between yaks (Bos grunniens) and Qaidam cattle (Bos taurus) fed a low protein diet with different energy levels.

The yak (Bos grunniens), an indigenous bovine on the Qinghai-Tibetan plateau (QTP), is reported to digest low quality forage to a greater extent and to require less protein and energy for maintenance than the introduced Qaidam cattle (Bos taurus). Ruminal bacteria play a major role in feed degradation, and therefore, we hypothesized that ruminal bacteria composition would differ between yaks and cattle, and confer an advantage to yaks for poor quality diets. To test our hypothesis, we determined the ruminal bacteria profiles, rumen fermentation parameters, and enzyme activities in these bovine species consuming a low-protein diet differing in energy level. Six castrated yaks (155 ± 5.8 kg) and 6 castrated Qaidam cattle (154 ± 8.0 kg) were used in two concurrent 4 × 4 Latin square designs with 2 additional animals of each species in each period. The animals were offered a low-protein diet of 70.4 g/kg dry matter (DM) and one of four metabolizable energy levels, namely 6.62, 8.02, 9.42, and 10.80 MJ/kg. Ruminal pH, concentrations of ammonia-N and total volatile fatty acids (VFAs), the molar proportion of acetate, and the ratio of acetate to propionate (A:P) were greater (P < 0.05), whereas the molar proportion of propionate was lesser (P = 0.043) in yaks than in cattle. With increasing dietary energy level, ruminal pH, the molar proportion of acetate and the ratio of A:P decreased linearly (P < 0.05), whereas, the concentration of total VFAs, molar proportions of propionate, butyrate, iso-butyrate, and iso-valerate and concentration of ammonia-N increased linearly (P < 0.05). The relative abundance (RA) of Firmicutes increased linearly (P < 0.01), whereas, the RA of Bacteroidetes decreased linearly (P < 0.001) with increasing energy level in both bovine species. The RAs of Prevotella and Rikenellaceae_RC9_gut_group decreased linearly (P < 0.05) with increasing energy level in both yaks and cattle. The RAs of fibrolytic (e.g., Rikenellaceae_RC9_gut_group), and H2-incorporating (e.g., Quinella) bacteria were greater (P < 0.05) in yaks than in cattle. We concluded that the two bovines differ in ruminal bacterial profiles and rumen fermentation parameters, and confer an advantage to yaks over cattle in consuming a low protein diet with differing energy level.

Effect of Clostridium butyricum Supplementation on in vitro Rumen Fermentation and Microbiota With High Grain Substrate Varying With Media pH Levels.

Clostridium butyricum (C. butyricum) can survive at low pH, and it has been widely used as an alternative to antibiotics for the improvement of feed efficiency and animal health in monogastrics. A recent study suggested that the improved ruminal fermentation with supplementing C. butyricum is may be associated with increasing the abundance of rumen microbiota in Holstein heifers, as ruminal pH plays a key role in rumen microbiota and the probiotics are often active in a dose-dependent manner. The objective of this study was to determine the effects of increasing the doses of C. butyricum on gas production (GP) kinetics, dry matter disappearance (DMD), fermentation characteristics, and rumen microbiota using a high grain substrate in batch culture varying with media pH levels. The doses of C. butyricum were supplemented at 0 (control), 0.5 × 106, 1 × 106, and 2 × 106 CFU/bottle, respectively, at either media pH 6.0 or pH 6.6. The fermentation microbiota at 0 and 1 × 106 CFU/bottle were determined using the 16S rRNA high throughput sequencing technology. Overall, the GP, DMD, total volatile fatty acid (VFA) concentration, and the ratio of acetate:propionate were higher (P <0.01) at media pH 6.6 than at pH 6.0. However, there was interaction between pH × dose of C. butyricum for rate constant of GP (P = 0.01), average GP rate (P = 0.07), and volume of GP (P = 0.06); with the increase in C. butyricum supplementation, the GP kinetics were not changed at media pH 6.0, but the volume (P = 0.02), rate of GP (P = 0.01), and average GP rate (P = 0.01) were quadratically changed at media pH 6.6. The DMD was not affected by increasing the supplementation of C. butyricum. The molar proportions of propionate (P <0.09), butyrate (P <0.06), and NH3-N concentration (P = 0.02) were quadratically changed with increasing supplementation of C. butyricum regardless of media pH levels. The interactions between media pH level and dose of C. butyricum supplementation were noticed for alpha diversity indexes of Shannon (P = 0.02) and Evenness (P = 0.04). The alpha diversity indexes increased (P <0.05) except for Chao1 with supplementation of C. butyricum. The unweighted uniFrac analysis showed that the group of control at media pH 6.0 and control at media pH 6.6, and supplementation of C. butyricum and control at media pH 6.0 clustered separately from each other. At the phylum level, relative abundance (RA) of Bacteroidota was lower (P <0.01) and Firmicutes was higher (P <0.01) at media pH 6.6 than pH 6.0. Moreover, RA of Proteobacteria decreased (P <0.05) with supplemented C. butyricum at either media pH 6.6 or pH 6.0. At media pH 6.6, RA of Rikenellaceae_RC9_gut_group and Prevotella were decreased, and CAG-352 was increased (at genus level) compared to pH 6.0. Supplementation of C. butyricum decreased RA of Rikenellaceae_RC9_gut_group and increased CAG-352 at media pH 6.0. It could hence be concluded that manipulating media pH level and supplementation of C. butyricum effectively modulated in vitro rumen fermentation characteristics and microbiota but in a dose depending manner of C. butyricum addition.

Co-expression of transcription factors ZmC1 and ZmR2 establishes an efficient and accurate haploid embryo identification system in maize.

Because of their high efficiency during chromosome doubling, immature haploid maize (Zea mays L.) embryos are useful for doubled haploid production. The R1-nj marker is commonly used in doubled haploid breeding and has improved the efficiency of haploid identification. However, its effectiveness is limited by genetic background and environmental factors. We addressed this technical challenge by developing an efficient and accurate haploid embryo identification marker through co-expression of two transcription factor genes (ZmC1 and ZmR2) driven by the embryo-aleurone-specific bidirectional promoter PZmBD1 ; these factors can activate anthocyanin biosynthesis in the embryo and aleurone layer during early seed development. We developed a new haploid inducer, Maize Anthocyanin Gene InduCer 1 (MAGIC1), by introducing the transgenes into the haploid inducer line CAU6. MAGIC1 could identify haploids at 12 days after pollination, which is nine days earlier than CAU6. Importantly, MAGIC1 increased haploid identification accuracy to 99.1%, compared with 88.3% for CAU6. In addition, MAGIC1 could effectively overcome the inhibition of anthocyanin synthesis in some germplasms. Furthermore, an upgraded anthocyanin marker was developed from ZmC1 and ZmR2 to generate MAGIC2, which could identify haploids from diploids due to differential anthocyanin accumulation in immature embryos, coleoptiles, sheaths, roots, leaves, and dry seeds. This haploid identification system is more efficient and accurate than the conventional R1-nj-based method, and it simplifies the haploid identification process. Therefore, this system provides technical support for large-scale doubled haploid line production.

Effect of commercial slow-release urea product on in vitro rumen fermentation and ruminal microbial community using RUSITEC technique.

BACKGROUND: The objectives of this study were to determine the effect of commercial slow-release urea (SRU) on in vitro fermentation characteristics, nutrient digestibility, gas production, microbial protein synthesis and bacterial community using a rumen simulation technique (RUSITEC). The experiment was a completely randomized design with four treatments and four replications of each treatment. Treatments were: control diet (no SRU addition), control diet plus 0.28% SRU (U28), or plus 0.56% SRU (U56), and control diet that was modified substituting a part of soybean meal equivalent to 0.35% SRU (MU35; dry matter [DM] basis). The experiment consisted of 8 d of adaptation and 7 d of data and sample collection. Rumen inoculum was obtained from three ruminally fistulated Angus cows fed the same diet to the substrate incubated. RESULTS: Digestibility of DM, organic matter (OM), crude protein (CP), fibre and starch was not affected, but daily production of gas (P < 0.07) and methane (P < 0.05) was quadratically increased with increasing SRU supplementation. The increase of SRU addition did not affect fermentation pH and total volatile fatty acid (VFA) production, whereas linearly (P < 0.01) decreased proportion of propionate, and linearly (P < 0.01) increased acetate to propionate ratio and ammonia nitrogen (N) concentration. The microbial N efficiency was also linearly (P < 0.03) improved with increasing supplementation of SRU. In comparison with control diet, the dietary substitution of SRU for part of soybean meal increased (P < 0.05) the digestibility of DM, OM and CP and decreased (P < 0.02) the total gas production. The total VFA production and acetate to propionate ratio did not differ between control and MU35, whereas the proportion of butyrate was lower (P < 0.05) and that of branched-chain VFA was greater (P < 0.05) with MU35 than control diet. Total and liquid-associated microbial N production as well as ammonia N concentration were greater (P < 0.03) with MU35 than control diet. Observed operational taxonomic units (OTUs), Shannon diversity index, and beta diversity of the microbial community did not differ among treatments. Taxonomic analysis revealed no effect of adding SRU on the relative abundance of bacteria at the phylum level, while at the genus level, the beneficial impact of SRU on relative abundance of Rikenellaceae and Prevotellaceae in feed particle-associated bacteria, and the abundance of Roseburia in liquid associate bacteria was greater (P < 0.05) with MU35. CONCLUSIONS: Supplementation of a dairy cow diet with SRU showed potential of increase in ammonia N concentration and microbial protein production, and change fermentation pattern to more acetate production. Adding SRU in dairy cow diet also showed beneficial effect on improving digestibility of OM and fibre. The results suggest that SRU can partially substitute soybean meal in dairy cow diet to increase microbial protein production without impairing rumen fermentation.

Nitrogen excretion from beef cattle fed a wide range of diets compiled in an intercontinental dataset: a meta-analysis.

Manure N from cattle contributes to nitrate leaching, nitrous oxide, and ammonia emissions. Measurement of manure N outputs on commercial beef cattle operations is laborious, expensive, and impractical; therefore, models are needed to predict N excreted in urine and feces. Building robust prediction models requires extensive data from animals under different management systems worldwide. Thus, the study objectives were to 1) collate an international dataset of N excretion in feces and urine based on individual observations from beef cattle; 2) determine the suitability of key variables for predicting fecal, urinary, and total manure N excretion; and 3) develop robust and reliable N excretion prediction models based on individual observation from beef cattle consuming various diets. A meta-analysis based on individual beef data from different experiments was carried out from a raw dataset including 1,004 observations from 33 experiments collected from 5 research institutes in Europe (n = 3), North America (n = 1), and South America (n = 1). A sequential approach was taken in developing models of increasing complexity by incrementally adding significant variables that affected fecal, urinary, or total manure N excretion. Nitrogen excretion was predicted by fitting linear mixed models with experiment as a random effect. Simple models including dry matter intake (DMI) were better at predicting fecal N excretion than those using only dietary nutrient composition or body weight (BW). Simple models based on N intake performed better for urinary and total manure N excretion than those based on DMI. A model including DMI and dietary component concentrations led to the most robust prediction of fecal and urinary N excretion, generating root mean square prediction errors as a percentage of the observed mean values of 25.0% for feces and 25.6% for urine. Complex total manure N excretion models based on BW and dietary component concentrations led to the lowest prediction errors of about 14.6%. In conclusion, several models to predict N excretion already exist, but the ones developed in this study are based on individual observations encompassing larger variability than the previous developed models. In addition, models that include information on DMI or N intake are required for accurate prediction of fecal, urinary, and total manure N excretion. In the absence of intake data, equations have poor performance as compared with equations based on intake and dietary component concentrations.