Do cows with stereotypic tongue-rolling behaviour cope better with their environment?

Introduction: Stereotypic behaviours, especially oral stereotypic behaviours, are frequently expressed in farm animals. Tongue-rolling is the most common oral stereotypic behaviour in dairy cows (Bos taurus). If animals frequently display stereotypic behaviours, this is an indication of poor welfare. It has been suggested that animals express stereotypic behaviours as a way of coping with stress. As a result, animals with stereotypic behaviours may have lower levels of stress hormones than animals without stereotypic behaviours. Methods: In this study, 916 Holstein cows in the first lactation were subjected to scan sampling behavioural observations 200 times for 10 days. All cows were assigned to either a stereotypic behaviours group (SB) or a control group (CON). The SB group was further subdivided into a tongue-rolling group (TR) and an other-stereotypic behaviours group (OS). The TR group was also split into an only tongue-rolling group (OTR) and a mixed tongue-rolling and other stereotypic behaviours group (TROS). Some cows in the TR group belonged to an extreme tongue-rolling group (ETR). Hair and saliva samples were collected from 601 cows to test cortisol concentrations and dairy herd improvement (DHI) data were collected from a total of 762 cows. Results: There were no differences in hair or saliva cortisol concentrations between the groups (p>0.05), and the frequencies of tongue-rolling were not associated with cortisol concentrations (p>0.05). For DHI in cows, the milk protein percentage (p = 0.028), milk true protein percentage (p = 0.021) and milk crude protein percentage (p = 0.023) of cows in the ETR group were significantly lower than those in the CON group. For cows in ETR group, as the frequencies of tongue-rolling increased, the milk protein percentage (p = 0.034, r = 0.365), milk true protein percentage (p = 0.022, r = 0.393) and milk crude protein percentage (p = 0.035, r = 0.363) increased. Discussion: We investigated the relationship between stereotypic behaviours and stress by using a non-invasive sampling method to minimise harm to the cows. We suggest that tongue-rolling may not be a way for cows to cope with stress, at least in terms of cortisol concentrations.

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.

Impact of Heat Stress on Blood, Production, and Physiological Indicators in Heat-Tolerant and Heat-Sensitive Dairy Cows.

Heat stress affects production and health in cows severely. Since it is difficult to define heat-tolerant animals, studies of response to heat stress are important for understanding dairy cows' health and production. However, information on the impact of heat stress on various indicators in heat-tolerant and heat-sensitive cows is sparse. This study aimed to investigate the effects of heat stress (HS) on blood, production, and physiological indicators in heat-tolerant and heat-sensitive cows. A total of 43 dairy cows were used from 9 May to 7 August 2021, under Temperature-Humidity Index (THI) measurements that ranged from 65.9 to 86.7. We identified cows that were tolerant or sensitive to HS based on the slope of the response of physiological and production traits against THI during the HS period by using a clustering method. After HS, serum glucose (Glu), cortisol (COR), 5-hydroxytryptamine (5-HT), and interleukin-6 (IL-6) levels of cows in the heat-tolerant group were lower than in the heat-sensitive group (p < 0.05). With THI as the predictor, the R2 for predicting respiration rate (RR) and body surface temperature (BT) in heat-tolerant cows was 0.15 and 0.16, respectively, whereas the R2 for predicting RR and BT in heat-sensitive cows was 0.19 and 0.18, respectively. There were low to moderate, positive correlations between RR, BT, and MY with THI, with Pearson correlation coefficients ranging from r = 0.11 to 0.4 in the heat-tolerant group, and from r = 0.24 to 0.43 in the heat-sensitive group. There was a low positive correlation between VT and THI, with a Spearman correlation coefficient of r = 0.07 in the heat-sensitive group. The heat-tolerant dairy cows had lower MY losses and had lower MY (p = 0.0007) in mixed models. Heat-tolerant cows with low-stress levels, through upregulating RR rapidly, increased their adaptability to thermal environments. They have better thermoregulation capability; the hypothalamic-pituitary-adrenal (HPA) axis regulated the thermoregulatory in animals by releasing a variety of neurotransmitters and hormones.

Ubiquitylome study reveals the regulatory effect of α-lipoic acid on ubiquitination of key proteins in tryptophan metabolism pathway of pig liver.

The decline in antioxidant defenses make it easily for human and animals to suffer from liver damage and diseases induced by oxidative stress, causing enormous losses to human health and livestock production. As one of the canonical protein post-translational modifications (PTMs), ubiquitination is widely involved in cell proliferation, apoptosis and damage/repair response, and is proven to be involved in the ability of mammals to resist oxidative stress. To explore whether α-lipoic acid (LA), a safe and efficient antioxidant, plays a role in regulating liver antioxidant status by PTMs, proteins in livers of pigs fed with LA were analyzed at the level of proteome and ubiquitylome. Based on proteome-wide enrichment of ubiquitination, a total of 7274 proteins were identified and 5326 were quantified, we also identified 1564 ubiquitination sites in 580 ubiquitinated proteins, among which there were 136 differentially ubiquitinated sites in 103 differentially ubiquitinated proteins upon LA. Further bioinformatics analysis showed that these differential proteins were mainly enriched in tryptophan metabolic pathway, and accompanied by significantly improvement of liver antioxidant capacity. We revealed the regulatory effect of LA on ubiquitination of kynurenine 3-monooxygenase (KMO) and other key proteins in tryptophan metabolism pathway of pig liver for the first time.

GRP94 Inhabits the Immortalized Porcine Hepatic Stellate Cells Apoptosis under Endoplasmic Reticulum Stress through Modulating the Expression of IGF-1 and Ubiquitin.

Endoplasmic reticulum stress (ERS) is closely related to the occurrence and progression of metabolic liver disease. The treatment targeting glucose-regulated protein 94 (GRP94) for liver disease has gotten much attention, but the specific effect of GRP94 on hepatocyte apoptosis is still unclear. So far, all the studies on GRP94 have been conducted in mice or rats, and little study has been reported on pigs, which share more similarities with humans. In this study, we used low-dose (LD) and high-dose (HD) tunicamycin (TM) to establish ERS models on piglet livers and immortalized porcine hepatic stellate cells (HSCs). On the piglet ERS model we found that ERS could significantly (p < 0.01) stimulate the secretion and synthesis of insulin-like growth factor (IGF-1), IGF-1 receptor (IGF-1R), and IGF-binding protein (IGFBP)-1 and IGFBP-3; however, with the increase in ERS degree, the effect of promoting secretion and synthesis significantly (p < 0.01) decreased. In addition, the ubiquitin protein and ubiquitination-related gene were significantly increased (p < 0.05) in the LD group compared with the vehicle group. The protein level of Active-caspase 3 was significantly increased (p < 0.01) in the HD group, however, the TUNEL staining showed there was no significant apoptosis in the piglet liver ERS model. To explore the biofunction of ER chaperone GRP94, we used shRNA to knock down the expression of GRP94 in porcine HSCs. Interestingly, on porcine HSCs, the knockdown of GRP94 significantly (p < 0.05) decreased the secretion of IGF-1, IGFBP-1 and IGFBP-3 under ERS, but had no significant effect on these under normal condition, and knockdown GRP94 had a significant (p < 0.01) effect on the UBE2E gene and ubiquitin protein from the analysis of two-way ANOVA. On porcine HSCs apoptosis, the knockdown of GRP94 increased the cell apoptosis in TUNEL staining, and the two-way ANOVA analysis shows that knockdown GRP94 had a significant (p < 0.01) effect on the protein levels of Bcl-2 and Caspase-3. For CCK-8 assay, ERS had a significant inhibitory(p < 0.05) effect on cell proliferation when treated with ERS for 24 h, and both knockdown GRP94 and ERS had a significant inhibitory(p < 0.05) effect on cell proliferation when treated with ERS for 36 h and 48 h. We concluded that GRP94 can protect the cell from ERS-induced apoptosis by promoting the IGF-1 system and ubiquitin. These results provide valuable information on the adaptive mechanisms of the liver under ERS, and could help identify vital functional genes to be applied as possible diagnostic biomarkers and treatments for diseases induced by ERS in the future.

Current Attitudes of Chinese Dairy Practitioners to Pain and Its Management in Intensively Raised Dairy Cattle.

Pain in dairy cattle is gaining attention globally. This study investigated the current attitudes of Chinese dairy practitioners to pain and its management in intensively raised dairy cattle. A total of 465 valid questionnaires with 26 painful conditions scored on numerical rating scales were collected from dairy practitioners. Data were analysed by descriptive statistics, analysis of variance, principal component analysis, and multivariate regression models. Dystocia was perceived as the most painful, while mild mastitis with milk changes only was perceived as the least painful. Respondents who agreed with the statement "pain management is worthwhile" tended to give a higher pain score. Young respondents (≤23 years old) and those from farms with ≤1000 cattle had lower pain scores for conditions with severe pain and low variability but higher pain scores for conditions with less severe pain and high variability, whereas highly educated respondents had consistently lower pain scores. As for pain management, older respondents (≥24 years old) tended to choose non-steroidal anti-inflammatory drugs, and farms with >1000 cattle were more likely to use analgesics. Training in pain perception and management should be emphasised with the hope of promoting animal welfare and reducing unnecessary production losses.

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.

4-tert-butylphenol triggers common carp hepatocytes ferroptosis via oxidative stress, iron overload, SLC7A11/GSH/GPX4 axis, and ATF4/HSPA5/GPX4 axis.

4-tert-butylphenol (4-tBP) is a toxic environmental pollutant with moderate bioaccumulation, environmental persistence, and long-term toxicity. Its toxicity to aquatic organisms has become an issue of concern. However, the molecular mechanism of 4-tBP toxicity to aquatic organisms remained unclear. Liver is a target organ for environmental pollutants. Here, we established 4-tBP-exposed toxicity model in vivo and primary hepatocyte model in vitro in common carp (Cyprinus carpio L.). We found increased hepatic-somatic index (HSI) and abnormal serum biochemical indexes (ALT, AST, and LDH) after 4-tBP exposure, indicating liver damage. We further revealed that 4-tBP damaged the structural integrity of the livers with typical features of ferroptosis. Based on toxicogenomics analysis, we found ferroptosis is likely to be involved in the mechanism of 4-tBP-induced liver damage. Moreover, our in vivo and in vitro experiment provided evidences that 4-tBP-exposure led to excess oxidative stress, iron overload, decreased MMP, and abnormal expression of ferroptosis-related factors. Interestingly, ferrostatin-1 (Fer-1, a ferroptosis inhibitor) pretreatment alleviated above changes. In summary, we demonstrated that 4-tBP triggered hepatocytes ferroptosis via oxidative stress, iron overload, SLC7A11/GSH/GPX4 axis, and ATF4/HSPA5/GPX4 axis. For the first time, we discovered that Fer-1 can ameliorate the toxicity of 4-tBP, which needs more investigations. Our results provided a scientific basis of molecular mechanism of 4-tBP-induced fish poisoning.

Acute Endoplasmic Reticulum Stress Induces Inflammation Reaction, Complement System Activation, and Lipid Metabolism Disorder of Piglet Livers: A Proteomic Approach.

Endoplasmic reticulum stress (ERS) is closely associated with the occurrence and development of many liver diseases. ERS models mostly include experimental animals such as rats and mice. However, pigs are more similar to humans with regards to digestion and metabolism, especially liver construction, yet few reports on ERS in pigs exist. In order to explore changes in the liver under ERS, we used tunicamycin (TM), which can cause liver jaundice and damage liver function, to establish acute ERS models in piglets using a low TM dosage (LD, 0.1 mg/kg body weight (bw)), high TM dosage (HD, 0.3 mg/kg bw), or vehicle for 48 h. We found that both LD- and HD-induced ERS, as verified by the ERS-linked proteins. Furthermore, the concentrations of the proinflammatory cytokines, namely, TNF-α and IL-6 were elevated in TM-treated piglet livers, and the plasma levels of IL-6 and CRP were also higher, indicating the occurrence of inflammation in TM-treated piglets. The complement system was activated in TM-treated piglets, as indicated by increased levels of complement factors and activation products C3, C5a, and AP50. In order to gain insights into the global changes in liver proteins under ERS, we performed an isobaric tag for relative and absolute quantitation (iTRAQ)-based proteomic analysis on the livers of HD- and vehicle-treated piglets. Proteomic analysis identified 311 differentially expressed proteins (DEPs) between the two groups, and a Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis suggested that the DEPs were mainly enriched in signaling pathways such as metabolic pathways, protein processing in the endoplasmic reticulum, and complement and coagulation cascades. Many proteins involved in protein folding, lipid transport, and oxidation were upregulated. Proteins involved in lipid synthesis were downregulated to alleviate liver steatosis, and most complement factors were upregulated to protect the body, and Pearson correlation analysis found that most of the DEPs in the complement and coagulation pathway were significantly correlated with plasma CRP, IL6 and AP50. Our results revealed that TM can activate ERS, marked by liver injury and steatosis, inflammatory reactions, and complement activation in piglets.

Aldehyde Dehydrogenase 2 as a Therapeutic Target in Oxidative Stress-Related Diseases: Post-Translational Modifications Deserve More Attention.

Aldehyde dehydrogenase 2 (ALDH2) has both dehydrogenase and esterase activity; its dehydrogenase activity is closely related to the metabolism of aldehydes produced under oxidative stress (OS). In this review, we recapitulate the enzyme activity of ALDH2 in combination with its protein structure, summarize and show the main mechanisms of ALDH2 participating in metabolism of aldehydes in vivo as comprehensively as possible; we also integrate the key regulatory mechanisms of ALDH2 participating in a variety of physiological and pathological processes related to OS, including tissue and organ fibrosis, apoptosis, aging, and nerve injury-related diseases. On this basis, the regulatory effects and application prospects of activators, inhibitors, and protein post-translational modifications (PTMs, such as phosphorylation, acetylation, S-nitrosylation, nitration, ubiquitination, and glycosylation) on ALDH2 are discussed and prospected. Herein, we aimed to lay a foundation for further research into the mechanism of ALDH2 in oxidative stress-related disease and provide a basis for better use of the ALDH2 function in research and the clinic.

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 a synthetic feline facial pheromone product on stress during transport in domestic cats: a randomised controlled pilot study.

OBJECTIVES: Transport can be a stressful experience for domestic cats. The objective of this study was to evaluate the effect of a new synthetic feline facial pheromone product on relieving stress in domestic cats during short-distance transport. METHODS: A randomised, blinded, placebo-controlled pilot study was conducted. Prior to baseline screening, cat owners were trained via online meetings in the assessment of stress-related behaviours and overall stress visual analogue scoring. All assessments were completed 30 mins after the start of transport. A total of 150 domestic cats exhibiting stress-related behaviours during baseline screening were recruited and randomly assigned to either the treatment group (n = 75) or the placebo group (n = 75). For the placebo-controlled intervention, the product and placebo, which were identical in appearance, were provided to the treatment and placebo groups, respectively. Fifteen minutes after spraying the carriers with the product or placebo, cat owners were required to take their cat out and complete the same assessments (baseline screening). RESULTS: After the intervention, the performance of the stress-related behaviours curling, lack of motion and meowing during transport outside the home was significantly reduced in the treatment group compared with the placebo group (P <0.05). When the baseline stress visual analogue scale (VAS) score was >2.94 cm, cats in the treatment group had significantly lower post-intervention stress VAS scores compared with those in the placebo group (P <0.05). CONCLUSIONS AND RELEVANCE: A synthetic feline facial pheromone product can alleviate short-distance transport-related stress in domestic cats with higher stress scores. This finding will benefit domestic cats during transport outside of the home.

Research Progress on Oxidative Stress and Its Nutritional Regulation Strategies in Pigs.

Oxidative stress refers to the dramatic increase in the production of free radicals in human and animal bodies or the decrease in the ability to scavenging free radicals, thus breaking the antioxidation-oxidation balance. Various factors can induce oxidative stress in pig production. Oxidative stress has an important effect on pig performance and healthy growth, and has become one of the important factors restricting pig production. Based on the overview of the generation of oxidative stress, its effects on pigs, and signal transduction pathways, this paper discussed the nutritional measures to alleviate oxidative stress in pigs, in order to provide ideas for the nutritional research of anti-oxidative stress in pigs.

Cadmium-induced Oxidative Stress and Immunosuppression Mediated Mitochondrial Apoptosis via JNK-FoxO3a-PUMA pathway in Common Carp (Cyprinus carpio L.) Gills.

Cadmium (Cd)-caused water environment pollution has become a matter of concern. Gill is an organ with respiratory and mucosal immune functions, and is also one of the organs directly attacked by pollutants. It was found that excess Cd could cause Cd accumulation and gill injury in carp. However, the mechanism of Cd-caused damage in common carp gills is still unclear. Oxidative stress, immunosuppression, and apoptosis took part in the mechanism of poisoning caused by some harmful substances. The aim of the study was to investigate complex molecular mechanism of apoptotic injury caused by Cd in common carp gills. Hence, in this study, we established a Cd poisoning model to explore whether excess Cd can induce apoptosis through observing histomorphology and apoptotic cells; and determining mineral elements, oxidative stress-related factors, immune-related, and apoptosis-related genes in common carp gills. Fifty-four fish were randomly separated into the control group and the Cd group and were cultured for 45 days. The water of the control group was drinking water and the water of the Cd group was CdCl2-added drinking water (0.26 mg/L Cd). In our results, we found that excess Cd increased Cd level, decreased the levels of essential mineral elements (Cu, Fe, Zn, and Mn), damaged mitochondria, and increased apoptotic cells in common carp gills, meaning that excess Cd caused Cd accumulation and apoptotic injury via mitochondrion in common carp gills. Furthermore, we found that Cd inhibited anti-apoptosis-related gene Bcl-2 and stimulated pro-apoptosis-related genes (JNK, FoxO3a, PUMA, Bax, Apaf-1, Caspase-9, and Caspase-3) on 15th, 30th, and 45th days. Above data meant that Cd exposure caused apoptosis via mitochondrion and JNK-FoxO3a-PUMA pathway in common carp gills. In addition, in our experiment, Cd treatment increased oxidants (H2O2 and MDA) and decreased antioxidants (CAT, GPx, GST, SOD, T-AOC, and GSH), indicating that Cd caused oxidative stress via oxidation/antioxidation imbalance. Meanwhile, compared to the control group, T-help 17 (Th17) cell-related factors (IL-17, TNF-α, and RORγ) were up-regulated, regulatory T (Treg) cell-related factors (IL-10 and Foxp3) were down-regulated, and IL-17/IL-10, TNF-α/IL-10, and RORγ/Foxp3 were increased in Cd-exposed group; meaning that excess Cd induced immunosuppression via the imbalance of Th17/Treg cells. Taken together, our findings indicated that JNK-FoxO3a-PUMA pathway and mitochondrion participated in oxidative stress and immunosuppression-mediated apoptosis caused by Cd in common carp (Cyprinus carpio L.) gills. Our data provided new perspectives on the negative effects of heavy metal pollutants on fish.

Short communication: The lag response of daily milk yield to heat stress in dairy cows.

Previous studies suggest that there exists a lag relationship between daily milk yield and heat stress. The values of heat stress indicators (e.g., temperature-humidity index and ambient temperature) before test day have a simple correlation with daily milk yield on test day. However, the simple correlation might not be the best description because daily milk yield and heat stress indicators have a nature of time series in common, and their correlations are cross correlations that could be affected by autocorrelations. We hope to give a more reliable estimation on the lag relationship of daily milk yield via excluding autocorrelations with transfer function modeling. In this study, we found a lag relationship between daily milk yield and heat stress indicators based on transfer function modeling. Heat stress indicators included ambient temperature and temperature-humidity index. The daily milk yield data from 123 cows were obtained during a consecutive 63-d period (July 10-September 10, 2016). The mean daily milk yield (MY) and the maximum daily ambient temperature (TA_max) satisfied the stationary hypothesis, and the cross correlation between them was calculated. Before excluding autocorrelation, MY at 0 to 4 d after test day had significant cross correlations with TA_max on test day. After excluding the influence of autocorrelations, MY at 1 to 3 d after the test day had significant cross correlations with TA_max on test day. This result suggested that MY would respond to TA_max 1 d after the test day. In addition, the strength of cross correlations between MY and TA_max decreased from 1 to 3 d in sequence, implying a declining lag response of MY that would last for 3 d. The transfer function model for MY and TA_max is written as: MYt = 16.90 + 0.74MYt- 1 - 0.25TA_maxt- 1 + Nt, where Nt is white noise. This model can be used to track and predict the dynamic response of MY to TA_max.

Metabolic Profiling of Plasma in Different Calving Body Condition Score Cows Using an Untargeted Liquid Chromatography-Mass Spectrometry Metabolomics Approach.

This study was undertaken to identify metabolite differences in plasma of dairy cows with a normal or high calving body condition score (CBCS), using untargeted liquid chromatography-mass spectrometry (LC-MS) metabolomics. Sixteen multiparous dairy cows were assigned to one of two groups based on CBCS (0 to 5 scale): Normal group (NBCS, 3.25 ≤ BCS ≤ 3.5, n = 8), and high BCS group (HBCS, BCS ≥ 4, n = 8). Plasma samples were collected for metabolomics analysis and evaluation of biomarkers of lipid metabolism (nonesterified fatty acid (NEFA) and ß-hydroxybutyrate (BHB)), and cytokines (leptin, adiponectin, tumor necrosis factor-α (TNF-α) and interleukin 6 (IL-6)). A total of 23 differential metabolites were identified, and functional analyses were performed using the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Among these metabolites, the concentrations of six lysophosphatidylcholines and one phosphatidylethanolamine, were lower in the HBCS group than in the NBCS group (p < 0.01). Furthermore, these metabolites were involved in these four pathways, among others: glycerophospholipid metabolism, retrograde endocannabinoid signaling, autophagy, and glycosylphosphatidylinositol (GPI)-anchor biosynthesis (p < 0.05). In addition, plasma concentrations of leptin (p = 0.06) and TNF-α (p = 0.08) tended to be greater while adiponectin (p = 0.09) lower in HBCS cows than in NBCS cows. The concentrations of NEFA, BHB, or IL-6 did not differ between NBCS and HBCS groups. More importantly, based on the results of the Spearman's correlation analysis, the seven important metabolites were negatively correlated with indices of lipid metabolisms, proinflammatory cytokines, and leptin, but positively correlated with adiponectin. These results demonstrate that CBCS has a measurable impact on the plasma metabolic profile, even when NEFA and BHB are not different. In addition, the identified differential metabolites were significantly correlated to lipid metabolism and inflammation in the over-conditioned fresh cows, which are expected to render a metabolic basis for the diseases associated with over-conditioned dry cows.

Predicting rectal temperature and respiration rate responses in lactating dairy cows exposed to heat stress.

Milk production and time effects are considered related to heat stress but they have not yet been combined in predictive models. In two parts, this study aimed to develop new models to predict heat stress (rectal temperature and respiration rate) of lactating dairy cows by inputting predictors, including ambient temperature (Ta), relative humidity (RH), wind speed (WS), milk yield (MY), and time blocks. In the first part of the study, we built the quantitative foundation for the second part, including the regression relation between respiration rate and rectal temperature (to convert predicted respiration rate to predicted body temperature), as well as between rectal temperature and respiration rate when heat stress was triggered (to recognize whether herds were under stress). In the second part, we built models that combined the abovementioned predictors to predict respiration rate. In part I, data were obtained from 45 high-producing Holstein cows within a Ta range of 9.5 to 30.8°C. We found a very strong correlation between mean respiration rate (MRR) and mean rectal temperature (MRT), where MRT = 0.021 × MRR + 37.6 (R2 = 0.925), suggesting that for each 4.8 breaths per minute (bpm) increase of MRR, MRT would be expected to increase by 0.1°C. Rectal temperature was determined to be 38.6°C when heat stress was triggered, which corresponded to a respiration rate of 48 bpm. In part II, data were obtained in 3 stalls within a Ta range of 6.9 to 33.3°C over 3 time blocks, all of which were the 90 min preceding milking (0630-0800, 1230-1400, and 1830-2000 h). We found a nonlinear response of MRR to Ta, which could be linearized by the quadratic term of Ta. The response of MRR was the highest in the 0630-0800 h block, followed by 1230-1400 h, and finally 1830-2000 h. We proposed a model combining 3 time blocks (R2 = 0.836): MRR in 0630-0800 h was determined to 56.28 + (-3.40 + 0.11 × Ta + 0.02 × RH) × Ta - 0.21 × RH - 2.82 × WS + 0.62 × MY; MRR in 1230-1400 h and 1830-2000 h were 4.6 and 10.3 bpm lower than that in 0630-0800 h, respectively (reducing the intercept of the expression in 0630-0800 h). Compared with temperature-humidity index equations, the proposed model performed better at suppressing prediction error, and had better sensitivity and accuracy in recognizing whether heat stress was triggered.

Artificial Grooming during Early Life could Boost the Activity and Human Affinity of Holstein Female Calves.

Early cow-calf separation management induced various welfare problems for dairy calves. We mimicked the maternal licking by manually brushing right after the Holstein female calves were born and during their first week of life, termed artificial grooming (AG). The behavior of these treated calves (AG, n = 17) was compared with the calves without artificial grooming (Con, n = 16) during daily behavioral observation around evening milk feeding and in the open field test (OFT) and novel human test (NHT). The number of calves ingesting starter on day six was recorded. The AG calves were observed to be more active and perform more oral behavior compared with the Con calves around evening milk feeding. In the OFT and NHT, the AG calves were again more active than the Con calves. Moreover, the AG calves tended to be less cautious and had more human interactions than the Con calves in the NHT. There tended to be a higher percentage of AG calves ingesting starter on day 6. In conclusion, artificial grooming during early life could boost the activity and the human affinity of female calves and it might advance their starter diet ingestion.

Effects of Different Farrowing Environments on the Behavior of Sows and Piglets.

We investigated the effect of different farrowing systems on the behavior of sows and piglets. In total, 22 hybrid sows (Yorkshire × Landrace) were randomly assigned into three farrowing systems, farrowing crate (FC), free farrowing pen with sloping walls (FFS), and free farrowing pen with sloping walls and nest materials (FFSN). The FFS and FFSN sows were more active, and exhibited less abnormal behaviors compared with the FC sows before and after parturition. FFS and FFSN piglets were more active compared with FC piglets. The increased activity of the FFS and FFSN sows might encourage the activity of their piglets, resulting in more proximity to sow behavior of their piglets. Providing nest materials improved the investigative behavior of sows and piglets. The total mortality of piglets in the free farrowing systems tended to be higher compared with the farrowing crate system. In conclusion, free farrowing system has beneficial effects on the behavior of sows and piglets but extra care in management needs to be taken to avoid piglet loss. Adding nest material in the farrowing pens is suggested to enrich the behavioral pattern of sows and piglets.

Identification of Metabonomics Changes in Longissimus Dorsi Muscle of Finishing Pigs Following Heat Stress through LC-MS/MS-Based Metabonomics Method.

Heat stress (HS) negatively affects meat quality by affecting material and energy metabolism, and exploring the mechanism underlying the muscle response to chronic HS in finishing pigs is important for the global pork industry. This study investigated changes in the metabolic profiles of the longissimus dorsi (LD) muscle of finishing pigs under high temperature using ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF-MS) and multivariate data analysis (MDA). Castrated male DLY pigs (Duroc × Landrance × Yorkshire pigs, n = 24) from 8 litters were divided into three treatment groups: constant optimal ambient temperature at 22 °C and ad libitum feeding (CR, n = 8); constant high ambient temperature at 30 °C and ad libitum feeding (HS, n = 8); and constant optimal ambient temperature 22 °C and pair-feeding to the control pigs (PF, n = 8). The metabolic profile data from LD muscle samples were analyzed by MDA and external search engines. Nine differential metabolites (L-carnosine, acetylcholine, inosinic acid, L-carnitine, L-anserine, L-α-glycerylphosphorylcholine, acetylcarnitine, thiamine triphosphate, and adenosine thiamine diphosphate) were involved in antioxidant function, lipid metabolism, and cell signal transduction, which may decrease post mortem meat quality and play important roles in anti-HS. Four metabolites (L-carnosine, acetylcholine, inosinic acid, and L-carnitine) were verified, and it was indicated that the muscle L-carnitine content was significantly lower in HS than in CR (p < 0.01). The results show that constant HS affects the metabolites in the LD muscle and leads to coordinated changes in the endogenous antioxidant defense and meat quality of finishing pigs. These metabonomics results provide a basis for researching nutritional strategies to reduce the negative effects of heat stress on livestock and present new insights for further research.