Metagenomic analysis reveals a dynamic rumen microbiome with diversified adaptive functions in response to dietary protein restriction and re-alimentation.

Understanding the dynamics of the rumen microbiome is crucial for optimizing ruminal fermentation to improve feed efficiency and addressing concerns regarding antibiotic resistance in the livestock production industry. This study aimed to investigate the adaptive effects of microbiome and the properties of carbohydrate-active enzymes (CAZy) and antibiotic resistance genes (ARGs) in response to dietary protein shifts. Twelve Charolais bulls were randomly divided into two groups based on initial body weight: 1) Treatment (REC), where the animals received a 7 % CP diet in a 4-week restriction period, followed by a 13 % CP diet in a 2-week re-alimentation period; 2) Control (CON), where the animals were fed the 13 % CP diet both in the restriction period and the re-alimentation period. Protein restriction decreased the concentrations of acetate, propionate, isovalerate, glutamine, glutamate, and isoleucine (P < 0.05), while protein re-alimentation increased the concentrations of arginine, methionine sulfoxide, lysine, and glutamate (P < 0.05). Protein restriction decreased the relative abundances of Bacteroidota but increased Proteobacteria, with no difference observed after re-alimentation. Protein restriction decreased relative abundances of the genera Bacteroides, Prevotella, and Bifidobacterium. Following protein recovery, Escherichia was enriched in CON, while Pusillibacter was enriched in REC, indicating that distinct microbial adaptations to protein shifts. Protein restriction increased GH97 while reducing GH94 and GT35 compared to CON. Protein restriction decreased abundances of KO genes involved in VFA production pathways, while they were recovered in the re-alimentation period. Protein restriction reduced tet(W/32/O) abundances but increased those of tet(X), nimJ, and rpoB2. Following protein re-alimentation, there was a decrease in ErmQ and tet(W/N/W), and an increase in Mef(En2) compared to CON, highlighting the impact of dietary protein on the distribution of antibiotic-resistant bacteria. Overall, comprehensive metagenomic analysis reveals the dynamic adaptability of the microbiome in response to dietary shifts, indicating its capacity to modulate carbohydrate metabolism and ARGs in response to protein availability.

Impact of dietary supplementation with ß-alanine on the rumen microbial crude protein supply, nutrient digestibility and nitrogen retention in beef steers elucidated through sequencing the rumen bacterial community.

This study investigated the effects of ß-alanine (ß-Ala) on rumen fermentation, nutrient digestibility, nitrogen (N) metabolism, plasma biochemical parameters, and rumen bacterial communities in beef steers. Six steers with initial liveweight of 252.8 ± 5.2 kg and 3 treatments of supplementing with 0, 30, or 60 g ß-Ala per day to basal diet were allocated in a replicated 3 × 3 Latin square design. Each experimental period was 20 d, of which the first 15 d were for adaptation and the subsequent 5 d were for sampling. The results showed that ß-Ala linearly increased the ruminal concentration of microbial crude protein (MCP) (P = 0.005), but it did not affect the ruminal concentrations of ammonia N and total volatile fatty acids (P > 0.10). ß-Ala also linearly increased the dry matter (DM) (P = 0.009), organic matter (OM) (P = 0.017) and crude protein (CP) (P = 0.043) digestibility, tended to decrease the acid detergent fiber digestibility (P = 0.077), but it did not affect the neutral detergent fiber digestibility (P = 0.641). ß-Ala quadratically increased the relative abundance of ruminal Bacteroidota (P = 0.021) at the phylum level, and increased Prevotella (P = 0.028) and Prevotellaceae_UCG-003 (P = 0.014), and decreased the relative abundance of NK4A214_group (P = 0.009) at the genus level. Feeding steers with ß-Ala linearly increased the urinary N (P = 0.006), urea excretions (P = 0.002) and the N retention (P = 0.004), but it did not affect the N utilization efficiency (P = 0.120). ß-Ala quadratically increased the plasma concentration of the total antioxidant capacity (P = 0.011) and linearly increased the plasma concentration of insulin-like growth factor-1 (P < 0.001). In summary, dietary supplementation with ß-Ala improved the rumen MCP supply and increased the digestibility of DM, OM, CP and the N retention. Further research is necessary to verify the ruminal degradability of ß-Ala and to investigate the mechanism of the impact of absorbed ß-Ala on the anti-oxidative ability in steers.

Immune Checkpoint Inhibitors Serve as the First-Line Treatment for Advanced Head and Neck Cancer.

OBJECTIVES: Immune checkpoint inhibitor (ICI) therapy has demonstrated substantial benefits for certain patients. We try to evaluate the merits and demerits of each immunotherapy to aid clinical treatment. METHODS: We conducted a comprehensive search of the PubMed, Embase, and Cochrane databases for randomized clinical trials published as of June 10, 2023. Our study included published clinical trials of ICI monotherapy or combination therapy, along with data on treatment-related adverse events (TRAE). Data regarding survival efficacy and adverse events of each randomized controlled trial (RCT) were collected. The Bayesian random effects model was utilized for the network meta-analysis (NMA). RESULTS: This study incorporated 19 RCTs, involving 5900 patients. Among 14 treatment regimens, Pembrolizumab combined with chemotherapy emerged as the most promising primary treatment for overall survival (OS) and objective response rate (ORR). Toripalimab combined with chemotherapy exhibited the highest likelihood of becoming the primary treatment for extending progression-free survival (PFS). Durvalumab showed the lowest probability of adverse events, suggesting a safer profile compared with other drugs. Camrelizumab combined with chemotherapy demonstrated a heightened risk of adverse events. Dual ICI Nivolumab/Ipilimumab surpassed Durvalumab/Tremelimumab in terms of ORR and adverse events. The standard of care (SOC) regimen did not exhibit strong performance across the four outcome indicators. CONCLUSION: Our analysis suggests that the integration of chemotherapy agents with ICIs enhances its efficacy as a first-line treatment for patients with advanced head and neck cancer (HNC). LEVEL OF EVIDENCE: 1 Laryngoscope, 134:749-761, 2024.

Automated recognition of the major muscle injury in athletes on X-ray CT images1.

BACKGROUND: In this research, imaging techniques such as CT and X-ray are used to locate important muscles in the shoulders and legs. Athletes who participate in sports that require running, jumping, or throwing are more likely to get injuries such as sprains, strains, tendinitis, fractures, and dislocations. One proposed automated technique has the overarching goal of enhancing recognition. OBJECTIVE: This study aims to determine how to recognize the major muscles in the shoulder and leg utilizing X-ray CT images as its primary diagnostic tool. METHODS: Using a shape model, discovering landmarks, and generating a form model are the steps necessary to identify injuries in key shoulder and leg muscles. The method also involves identifying injuries in significant abdominal muscles. The use of adversarial deep learning, and more specifically Deep-Injury Region Identification, can improve the ability to identify damaged muscle in X-ray and CT images. RESULTS: Applying the proposed diagnostic model to 150 sets of CT images, the study results show that Jaccard similarity coefficient (JSC) rate for the procedure is 0.724, the repeatability is 0.678, and the accuracy is 94.9% respectively. CONCLUSION: The study results demonstrate feasibility of using adversarial deep learning and deep-injury region identification to automatically detect severe muscle injuries in the shoulder and leg, which can enhance the identification and diagnosis of injuries in athletes, especially for those who compete in sports that include running, jumping, and throwing.

Evaluation of Holstein cows with different tongue-rolling frequencies: stress immunity, rumen environment and general behavioural activity.

BACKGROUND: The tongue-rolling behaviour of cows is regarded as an outward sign of stressed animals in a low welfare status. The primary aim of this observational study was to evaluate the association between the frequency of tongue-rolling behaviour and its physiological function. The secondary aim was to explore the relationship between general activities and the frequency of tongue-rolling behaviour of cows. A total of 126 scan sampling behavioural observations were collected over 7 d on 348 Holstein cows with the same lactation stage in the same barn. The tongue-rolling frequency was defined as the number of tongue-rolling observations as a percentage to the total observations per individual cow. According to their tongue-rolling frequency, the cows were grouped into the CON (no tongue-rolling), LT (frequency 1%), MT (frequency 5%), and HT (frequency 10%) groups. Six cows from each group were randomly selected for sampling. Serum samples, rumen fluid, milk yield, and background information were collected. The general behaviour data during 72 continuous hours of dairy cows, including eating time, rumination time, food time (eating time + rumination time), and lying time, were recorded by the collar sensor. RESULTS: Cortisol (P = 0.012), γ-hydroxybutyric acid (P = 0.008), epinephrine (P = 0.030), and dopamine (P = 0.047) levels were significantly higher in tongue-rolling groups than in the CON group. Cortisol levels and tongue-rolling frequency had a moderate positive correlation (linearly r = 0.363). With the increase in tongue-rolling frequency, the rumen pH decreased first and then increased (P = 0.013), comparing to the CON group. HT cows had significantly less food time than CON cows (P = 0.035). The frequency of tongue-rolling had a moderate negative relationship with rumination time (r = -0.384) and food time (r = -0.492). CONCLUSIONS: The tongue-rolling behaviour is considered as a passive coping mechanism, as the stress response in cows with high tongue-rolling frequency increased. Food intake and rumination activities were all closely related to the occurrence of tongue-rolling behaviour.

Effects of taurine on rumen fermentation, nutrient digestion, rumen bacterial community and metabolomics and nitrogen metabolism in beef steers.

BACKGROUND: The objectives of this study were to investigate the effects of taurine on rumen fermentation, rumen bacterial community and metabolomics, nitrogen metabolism and plasma biochemical parameters in beef steers. Six castrated Simmental steers (liveweight 402 ± 34 kg) and three levels of taurine (0, 20, 40 g d-1 ) were assigned in a replicated 3 × 3 Latin square design. Each experimental period included 15 days for adaptation and 5 days for sampling. RESULTS: Supplementing taurine did not affect the ruminal pH or concentrations of ammonia nitrogen and volatile fatty acids (P > 0.10), but linearly increased the ruminal concentrations of taurine (P < 0.001) and microbial crude protein (P = 0.041). Supplementing taurine linearly increased the neutral detergent fiber digestibility (P = 0.018), and tended to linearly increase dry matter digestibility (P = 0.095), tended to increase the fecal nitrogen excretion (P = 0.065) and increased the urinary taurine excretion (P < 0.001). Supplementing taurine quadratically increased the plasma concentration of triglycerides (P = 0.017), tended to linearly decrease growth hormone (P = 0.074), but did not affect other plasma parameters (P > 0.10). Supplementing taurine modified the rumen bacterial community and increased the ruminal concentration of taurine metabolite 2-hydroxyethoxysulfonic acid (P < 0.001). CONCLUSION: It was concluded that taurine improved ruminal microbial crude protein synthesis and increased fiber digestibility through modifying rumen bacterial community. It is necessary to clarify the ruminal hydrolysis of taurine in steers. © 2023 Society of Chemical Industry.

Feed tossing behaviour of Holstein cows: evaluation of physiological stress state and rumen fermentation function.

BACKGROUND: Abnormal or stereotyped behaviours in dairy cows are common in large-scale indoor farms and are usually accompanied by high physiological stress levels. Feed tossing is an abnormal behaviour commonly seen in cows while being fed, making farm management difficult. However, the reasons behind this behaviour have not been sufficiently reported. The objective of this study was to explore the changes in rumen fermentation, serum indicators, inflammatory conditions and the performance of cows with feed tossing behaviour. Holstein cows with similar lactation stages in the same barn were subjected to behaviour observations two times per day for 21 consecutive days. Ten cows with feed tossing behaviour (FT) and ten cows without abnormal behaviours (CON) were selected for further sampling. Plasma samples, rumen fluid, milk yield data of cows, and an indoor environment temperature-humidity index (THI) were collected. RESULTS: There was no significant difference in average daily milk yield during the observation period between feed-tossing cows (n = 68) and the other cows (n = 112). The number of cows showing FT behaviour had a moderately strong negative linear correlation with the THI of the environment. Compared to the CON cows, the FT cows had higher cortisol, norepinephrine and urea nitrogen levels in plasma, as well as higher plasma levels of inflammatory indicators, including total protein, lactate dehydrogenase, albumin, aspartate aminotransferase levels, and the ratio of aspartate aminotransferase to alanine aminotransferase. The FT cows had no significant variations from the CON cows regarding their rumen fermentation indicators, such as pH, ammonia nitrogen, and volatile fatty acids. In addition, 16S rRNA analysis revealed that there might be no clear association between the diversity and abundance of rumen bacteria and feed tossing behaviour. CONCLUSIONS: Our findings suggested that cows might have suffered from high levels of physiological stress and immune state for a long period when they exhibited FT behaviour. The environmental THI could affect the FT behaviour of cows; as the THI increases, the willingness of cows to throw decreases. This work provided the first evidence that feed tossing might be a response associated with high levels of physiological stress and immune. It also explored our insights into a commonly observed behavioural response to cow welfare traits.

Lipidomic profiling using GC and LC-MS/MS revealed the improved milk quality and lipid composition in dairy cows supplemented with citrus peel extract.

Eight lactating cows were used to determine the effects of citrus peel extract (CPE) on milk performance, antioxidant properties, and milk lipids composition. CPE supplementation up to 150 g/d (CPE150) increased milk yield and the proportions of unsaturated fatty acids of conjugated linoleic acid. CPE with abundant polyphenol and flavonoids can transfer these bioactive substances to mammary gland and improve the antioxidant properties of milk obtained from cows. Lipidomics revealed that 56 lipid species were altered between CON vs CPE150, and there were five key differential metabolic pathways. In particular, milk phosphatidylethanolamine and phosphatidylcholine were significantly increased with dietary CPE supplementation. In summary, our results provide insights into the modifications in the milk components and milk quality of dairy cows received CPE. The inclusion of CPE in the diet of dairy cows may be an effective and natural way to increase the antioxidant amounts and beneficial lipids in milk.

Potentials of using dietary plant secondary metabolites to mitigate nitrous oxide emissions from excreta of cattle: Impacts, mechanisms and perspectives.

Nitrous oxide (N2O) is a potent greenhouse gas as well as the key component depleting the ozone sphere of the earth. Cattle have high feed and water intakes and excrete large amounts of urine and feces. N2O can be produced from cattle excreta during storage and use as fertilizer. Mitigating the N2O emissions from cattle excreta during production is important for protecting the environment and the sustainable development of the cattle industry. Feeding cattle with low-protein diets increases N utilization rates, decreases N excretion and consequently reduces N2O emissions. However, this approach cannot be applied in the long term because of its negative impact on animal performance. Recent studies showed that dietary inclusion of some plant secondary metabolites such as tannins, anthocyanins, glucosinolates and aucubin could manipulate the N excretion and the urinary components and consequently regulate N2O emissions from cattle excreta. This review summarized the recent developments in the effects of dietary tannins, anthocyanins and glucosinolates on the metabolism of cattle and the N2O emissions from cattle excreta and concluded that dietary inclusion of tannins or anthocyanins could considerably reduce N2O emissions from cattle excreta.

Decreasing ruminal methane production through enhancing the sulfate reduction pathway.

Methane (CH4) production from ruminants accounts for 16% of the global greenhouse gas emissions and represents 2% to 12% of feed energy. Mitigating CH4 production from ruminants is of great importance for sustainable development of the ruminant industry. H2 is the primary substrate for CH4 production in the processes of ruminal methanogenesis. Sulfate reducing bacteria are able to compete with methanogens for H2 in the rumen, and consequently inhibit the methanogenesis. Enhancing the ruminal sulfate reducing pathway is an important approach to mitigate CH4 emissions in ruminants. The review summarized the effects of sulfate and elemental S on ruminal methanogenesis, and clarified the related mechanisms through the impacts of sulfate and elemental S on major ruminal sulfate reducing bacteria. Enhancing the activities of the major ruminal sulfate reducing bacteria including Desulfovibrio, Desulfohalobium and Sulfolobus through dietary sulfate addition, elemental S and dried distillers grains with solubles can effectively decrease the ruminal CH4 emissions. Suitable levels of dietary addition with different S sources for reducing the ruminal CH4 production, as well as maintaining the performance and health of ruminants, need to be investigated in the future.

Model for preventing tissue injury in the process of athlete training / Modelo para prevenção de lesões teciduais no processo de treinamento de atletas / Modelo para prevención de lesiones tisulares en el proceso de entrenamiento de atletas

ABSTRACT Introduction: The topic of preventing tissue damage during sports is a heavily debated one. Uncovering factors related to sports injuries is essential to deal with this issue. Objective: To analyze the causes of athletes' sports injuries, providing a theoretical basis for reducing the occurrence of sports injuries and increasing the success rate of athletes. Methods: This article uses statistical methods to investigate and analyze the causes of athletes' sports injuries. Results: Four important factors lead to athletes' sports injuries: poor physical fitness, inadequate warm-up activities, incorrect techniques, and excessive loads. Conclusion: Coaches should systematically prepare multi-year training plans, step by step, and pay attention to long-term effects. This is the only way athletes can achieve their best performance at the appropriate age. Level of evidence II; Therapeutic studies - investigation of treatment results.

RESUMO Introdução: A prevenção de danos teciduais durante a prática esportiva é um tópico acaloradamente debatido. Descobrir os fatores relacionados a ferimentos esportivos é essencial para lidar com esse agravo. Objetivo: Analisar as causas dos ferimentos esportivos em atletas, fornecendo uma base teórica para reduzir o número de ferimentos esportivos e aumentar a taxa de sucesso de atletas. Métodos: Esse artigo usa métodos estatísticos para investigar e analisar a causa das lesões esportivas em atletas. Resultados: Quatro fatores importantes levam às lesões esportivas dos atletas: preparo físico ruim, aquecimento inadequado, técnica incorreta e cargas excessivas. Conclusão: Técnicos deveriam preparar, passo-a-passo, treinamentos sistemáticos com duração de vários anos, levando em consideração seus efeitos de longo prazo. Só assim atletas podem atingir sua melhor performance na idade adequada Nível de evidência II; Estudos terapêuticos - investigação de resultados de tratamento.

RESUMEN Introducción: La prevención de daños tisulares durante la práctica deportiva es un tópico muy debatido. Descubrir los factores relacionados con las lesiones deportivas es esencial para lidiar con este problema. Objetivo: Analizar las causas de las lesiones deportivas en atletas, suministrando una base teórica para reducir el número de lesiones deportivas y aumentar la tasa de éxito de atletas. Métodos: Este artículo usa métodos estadísticos para investigar y analizar la causa de las lesiones deportivas en atletas. Resultados: Cuatro factores importantes ocasionan las lesiones deportivas de los atletas: preparación física insuficiente, calentamiento inadecuado, técnica incorrecta y cargas excesivas. Conclusión: Técnicos deberían preparar, paso a paso, entrenamientos sistemáticos con duración de varios años, teniendo en cuenta sus efectos a largo plazo. Solo así los atletas podrían alcanzar su mejor desempeño en la edad adecuada. Nivel de evidencia II; Estudios terapéuticos - investigación de resultados de tratamiento.

Physiological Indicators and Production Performance of Dairy Cows With Tongue Rolling Stereotyped Behavior.

Non-nutritive oral behaviors, especially tongue rolling, are prevalent in the stabled cow population. These behaviors mean that the environment or management process might not suit the cows, suggesting low welfare. However, few researches have reported the physiological indicators or production performance of dairy cows with the stereotyped behavior. This study aimed to determine physical conditions, daily activity, rumen fermentation, and milk production of cows with tongue-rolling behavior. Three hundred and fifty nine Holstein cows in the same barn and lactation stage were subjected to scan sampling behavior observations 126 times for 7 days. Ten cows with high-frequency tongue-rolling behavior (TON) and 10 cows without abnormal oral behavior (CON) were selected for further study. Serum sample, ruminal fluid, milk sample, and behavior record video of TON and CON cows were collected. TON cows had more drinking behavior and more stable lying behavior than the CON cows during the daytime. The body condition score of the TON cows decreased, while the milk yield, yield of milk fat, protein, and lactose in the study period increased. The TON cows had lower ruminal fluid pH, acetate/propionate ratio, and total volatile acid. The bacterial diversity in the ruminal fluid was not different between the two groups. Compared to CON cows, the TON cows had a higher level of serum stress indicators, such as cortisol, thyroid hormone, and norepinephrine, which positively correlated to the frequency of tongue-rolling behavior. Meanwhile, the TON cows had a higher level of lactate dehydrogenase, serum glucose, total triglyceride, total cholesterol, and Interleukin 6. Overall, it means they suffer from higher levels of stress and have higher energy metabolism for a long time when cows show tongue-rolling behavior. TON cows had suffered a higher stress level and had higher energy metabolic status for a long time. The TON cows might have better heat tolerance to the thermal environment by more lying and drinking time. Our data revealed the changes in milk production, physiological stress indicators of dairy cows with high-frequency tongue rolling behavior, which will provide essential knowledge for the in-depth understanding of tongue rolling behavior in dairy cows.

Effect of Grape Marc Added Diet on Live Weight Gain, Blood Parameters, Nitrogen Excretion, and Behaviour of Sheep.

A 39-day field experiment was conducted to assess the effect of grape marc (GM) feeding on sheep productivity, health, and environmental sustainability. Forty merino sheep were divided into two dietary groups, each having five replications (n = 4 sheep/replication). Experimental diet consisted of: (i) control: 55% lucerne hay + 40% wheat grain + 5% faba bean; (ii) GM treatment: control diet with 20% replaced by GM on a dry matter (DM) basis. The GM treatment contained 2-10% higher phytochemical contents than the control. The DMI from the GM treatment was 15% higher than the control (p < 0.001). No difference was found in sheep live weight gain, behaviour, and quality between groups (p > 0.05). No difference was found in total faecal production, faecal organic matter, and nitrogen contents (p > 0.05) and parasitic egg count. The GM treatment led to higher nitrogen intake (23.1 vs. 27.2 g/d) and faecal nitrogen excretion (6.3 vs. 8.7 g/d) compared to the control. Urinary creatinine, allantoin, and purine derivatives were lower in the GM treatment than control (p < 0.05). However, both groups had similar purine derivatives/DMI (i.e., indicator of rumen microbial protein synthesis efficiency; p > 0.05). Overall, the results showed that GM can replace 20% of the control ration to maintain sheep productivity, health, and environmental sustainability.

Effects of dietary inclusion with rapeseed cake containing high glucosinolates on nitrogen metabolism and urine nitrous oxide emissions in steers.

Two consecutive trials were conducted to investigate the effects of glucosinolates (GLS) in rapeseed cake (RSC) on nitrogen (N) metabolism and urine nitrous oxide (N2O) emissions in steers. In trial 1, 8 steers and 4 levels of RSC, i.e. 0, 2.7%, 5.4% and 8.0% dry matter (DM) (0, 6.0, 12.1, 18.1 µmol GLS/g DM) were allocated in a replicated 4 × 4 Latin square. In trial 2, the static incubation technique was used for measuring the N2O emissions of the urine samples collected from trial 1. The results of trial 1 indicated that dietary inclusion of RSC decreased the digested N and increased the fecal N excretion (P < 0.01), whereas it did not affect the urinary N excretion, total N excretion and N retention (P > 0.10). Dietary inclusion of RSC decreased the urinary excretion of urea while it increased allantoin, total purine derivatives, the predicted rumen microbial N flow and thiocyanate (SCN) (P < 0.05). Dietary inclusion of RSC did not affect the plasma triiodothyronine and thyroxine while it down-regulated the plasma relative concentrations of 4-aminohippuric acid, 3α,7α-dihydroxycoprostanic acid, phosphatidylserine (14:0/16:0), 6ß-hydroxyprogesterone, pyrrhoxanthinol, tatridin B, mandelonitrile rutinoside, taraxacoside (P < 0.05), and up-regulated hypoglycin B, neuromedin N (1-4), dhurrin, 5-deoxykievitone (P < 0.01). The results of trial 2 indicated that dietary RSC increased the steer urine N2O-N fluxes, the ratio of N2O-N to N application and the estimated steer urine N2O-N emissions (P < 0.01). A close correlation was found between the estimated steer urine N2O-N emissions and the output of urinary SCN (P < 0.001). In conclusion, dietary RSC increased the fecal N excretion, whereas it did not affect the urinary N excretion and the N retention rate in steers. Dietary RSC increased rather than decreased the urine N2O-N emissions even though it decreased the urinary excretion of urea. The SCN excreted in urine could be the major factor in increasing the urine N2O-N emissions. Whether other metabolites excreted into urine from RSC have an impact on the urine N2O-N emissions in steers needs to be investigated in the future.

Thiocyanate increases the nitrous oxide formation process through modifying the soil bacterial community.

BACKGROUND: Nitrous oxide (N2 O) is a potent greenhouse gas depleting the stratospheric ozone. Previous studies reported that the thiocyanate (TC) excretion in the urine of cattle fed rapeseed meals containing glucosinolates was positively correlated with the N2 O-nitrogen (N) emissions. The objectives of the experiment were to verify the effects and the mechanism of TC on the N2 O-N emissions from the soil applied with artificial urine using static incubation technique. Four levels of TC, that is 0.00, 0.26, 0.78 and 2.33 mmol L-1 were composited in artificial urine as experimental treatments. Soil inorganic N and bacterial community were analyzed to elucidate the effects of TC on the N2 O-N emissions of artificial urine. RESULTS: Adding TC increased the N2 O-N fluxes, the N2 O-N to N application ratio, and the estimated N2 O-N emissions from the soil applied with artificial urine both linearly and quadratically. The estimated N2 O-N emission (Y, in µmol) was increased with the TC adding level (X, in µmol) in a quadratic manner: Y = 52.57 + 4.47 X - 0.123 X 2 (R 2  = 0.70). Adding TC did not affect the soil bacterial diversity and richness, but increased the relative abundances of Nitrosomonas (both for nitrification and denitrification) and Hyphomicrobium, Lysobacter and Terrimonas (for denitrification), and tended to increase the relative abundances of denitrification and dissimilatory nitrate reduction to ammonium. CONCLUSION: TC increased the N2 O-N emissions of the soil applied with artificial urine possibly through enhancing the denitrification of nitrifiers in the soil. Field experiments are necessary to verify the laboratory results. © 2021 Society of Chemical Industry.

Effects of Dietary Supplementation with 2-Hydroxy-4-(methylthio)-butanoic Acid Isopropyl Ester as a Methionine Supplement on Nitrogen Utilization in Steers.

The objective of the experiment was to investigate the effects of dietary supplementation with 2-hydroxy-4-(methylthio)-butanoic acid isopropyl ester (HMBi) on the nitrogen (N) metabolism in beef steers. The plasma metabolites analyzed by metabolome profiling were used to clarify the impact mechanism. Three Simmental steers (body weight, 593 ± 23 kg) were used as experimental animals. Three levels of HMBi (i.e., 0, 12, and 24 g d-1) were added in a basal ration as experimental treatments. The steers and the dietary treatments were randomly allocated in a 3 × 3 Latin square design. The results showed that supplementing HMBi up to 24 g d-1 did not affect the N retention and N retention rate (NRR), and the fecal N/urinary N ratio even though it tended to linearly increase the uric acid N/urinary N ratio in steers. The results of plasma metabolome profiling showed that supplementing HMBi at 24 g d-1 upregulated the plasma concentrations of L-methionine (Met); Met-related metabolites including betaine, Met sulfoxide, and taurine; and L-isoleucine and tyrosine, whereas it downregulated L-serine, glycine, diaminopimelic acid, and other metabolites. The reason for the nonsignificant effect of HMBi on improving the N utilization in steers could be that the steers used in the experiment were in the fattening period. It is suggested to evaluate the effects of the dietary addition of HMBi using growing cattle in further research.

Adding polyethylene glycol to steer ration containing sorghum tannins increases crude protein digestibility and shifts nitrogen excretion from feces to urine.

The objectives of the experiment were to study the effects of adding polyethylene glycol (PEG) to steer ration containing high sorghum tannins on rumen fermentation, nutrient digestion, nitrogen (N) balance and plasma biochemical parameters. Eight growing steers at 16 months of age were allotted to a replicated 4 × 4 Latin square design with 4 treatments and 4 periods (19 d each). Polyethylene glycol at 0, 1.75, 3.50 and 7.00 g/kg dry matter (DM) were added to a basal ration containing 27.82% DM of sorghum grain (total tannins 3.3 g/kg DM) as the treatments. The results indicated that adding PEG quadratically increased the ruminal pH (P = 0.049), tended to linearly increase the ruminal concentration of total volatile fatty acids (P = 0.070), increased the molar proportion of acetate (P = 0.016), linearly decreased the molar proportion of butyrate (P = 0.015), and tended to increase the molar proportion of iso-valerate (P = 0.061) and the ruminal concentration of ammonia N (P = 0.092). Adding PEG tended to quadratically decrease the relative abundance of methanogenic archaea (P = 0.082), linearly decreased the relative abundance of Fibrobacter succinogenes (P = 0.008) and decreased the relative abundance of Butyrivibrio fibrisolvens (P = 0.048) at 7.00 g/kg DM. Dietary addition with PEG increased the crude protein (CP) digestibility (P < 0.001) and tended to increase the neutral detergent fiber digestibility (P = 0.066) in a linear manner. Adding PEG to basal ration also increased the plasma globulin concentration (P = 0.029) and tended to linearly increase the plasma total protein concentration (P = 0.069). Adding PEG linearly decreased the fecal N excretion (P < 0.001) and the fecal N-to-total N excretion ratio (P = 0.004) and increased the urinary N-to-total N excretion ratio (P = 0.004) and urinary urea excretion (P = 0.010) without affecting the urinary N and total N excretions (P > 0.05). It was concluded that adding PEG effectively improved the CP digestibility of the ration containing high sorghum tannins but increased the urinary urea excretion without improving the N retention and N retention rate in steers.

Comparison between the effects of feeding copper sulphate-treated and untreated rapeseed cake containing high glucosinolates on rumen fermentation, nutrient digestion and nitrogen metabolism in steers.

Two trials were carried out to study the effects of copper sulphate (CuSO4 ) on detoxifying glucosinolates (GLS) in rapeseed cake (RSC) and compare the effects of feeding CuSO4 -treated and untreated RSC on nutrient digestion and nitrogen (N) metabolism in steers. In Trial 1, different concentrations of CuSO4 solution (1.6 vs. 3.2 g CuSO4 ·5H2 O L-1 ), soaking temperatures (25 vs. 60°C) and drying methods (air drying at 60°C vs. freeze drying) were allocated in a 2 × 2 × 2 factorial arrangement in vitro. In Trial 2, six steers and dietary inclusions of untreated RSC (control), CuSO4 -treated RSC and CuSO4 -added RSC were assigned in a replicated 3 × 3 Latin square design. CuSO4 treatment in vitro decreased the contents of GLS and thiocyanate (TC) in RSC (p < 0.001). The total amount of GLS and TC decreased by 62.7-68.5% for all treatments. The animal trial showed that CuSO4 -treated RSC inclusion decreased ruminal concentration of valerate (p < 0.01), whereas it did not affect ruminal pH, ammonia N and total volatile fatty acids. Compared with the control, feeding CuSO4 -treated or CuSO4 -added RSC had no effect on plasma concentrations of triiodothyronine and thyroxine, N excretion and N retention. CuSO4 -treated RSC tended to increase neutral detergent fibre digestibility (p = 0.072) and urinary excretion of urea (p = 0.056). Urinary excretion of purine derivatives (p = 0.076) and rumen microbial N supply (p = 0.084) tended to decrease when feeding CuSO4 -treated RSC versus control. TC was found to be the only metabolite of GLS in rumen fluid, plasma and urine. It was feasible to detoxify GLS in RSC using low CuSO4 at room temperature. However, feeding CuSO4 -treated or CuSO4 -added RSC had minor effects on rumen fermentation, nutrient digestion and N metabolism in steers. CuSO4 treatment on RSC for feeding steers seems to be unnecessary.

Effects of different levels of rapeseed cake containing high glucosinolates in steer ration on rumen fermentation, nutrient digestibility and the rumen microbial community.

This trial was conducted to study the effects of dietary rapeseed cake (RSC) containing high glucosinolates (GLS) on rumen fermentation, nutrient digestion and the rumen microbial community in steers. Eight growing steers and four rations containing RSC (GLS 226·1 µmol/g DM) at 0·00, 2·65, 5·35 and 8·00 % DM were assigned in a replicate 4 × 4 Latin square design. The results indicated that increasing RSC levels increased the ruminal concentration of thiocyanate (SCN) (P < 0·01), decreased the ruminal concentration of ammonia nitrogen (NH3-N) and the molar proportion of isovalerate (P < 0·05), did not affect the ruminal concentration of total volatile fatty acids (P > 0·05), decreased the crude protein (CP) digestibility (P < 0·05) and increased the ether extract (EE) digestibility (P < 0·01). Increasing RSC levels tended to decrease the abundances of ruminal Ruminobacter amylophilus (P = 0·055) and Ruminococcus albus (P = 0·086) but did not affect methanogens, protozoa, fungi and other bacteria (P > 0·05). Increasing RSC levels in the ration did not affect the ruminal bacterial diversity (P > 0·05), but it increased the operational taxonomic units and the bacterial richness (P < 0·05) and affected the relative abundances of some bacteria at the phylum level and genus level (P < 0·05). In conclusion, RSC decreased the ruminal concentration of NH3-N and the CP digestibility, increased the EE digestibility and partly affected the ruminal bacterial community. SCN, as the metabolite of GLS, could be a major factor affecting these indices.

Steam Explosion Pretreatment Changes Ruminal Fermentation in vitro of Corn Stover by Shifting Archaeal and Bacterial Community Structure.

Steam explosion is an environment-friendly pretreatment method to improve the subsequent hydrolysis process of lignocellulosic biomass. Steam explosion pretreatment improved ruminal fermentation and changed fermentation pattern of corn stover during ruminal fermentation in vitro. The study gave a comprehensive insight into how stream explosion pretreatment shifted archaeal and bacterial community structure to change ruminal fermentation in vitro of corn stover. Results showed that steam explosion pretreatment dramatically improved the apparent disappearance of dry matter (DM), neutral detergent fiber (NDF), and acid detergent fiber (ADF). Steam explosion pretreatment significantly increased the molar proportion of propionate and decreased the ratio of acetate to propionate. At archaeal level, steam explosion pretreatment significantly increased the relative abundance of Methanobrevibacter, which can effectively remove metabolic hydrogen to keep the fermentation continuing. At bacterial level, the shift in fermentation was achieved by increasing the relative abundance of cellulolytic bacteria and propionate-related bacteria, including Spirochaetes, Elusimicrobia, Fibrobacteres, Prevotella, Treponema, Ruminococcus, and Fibrobacter.