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.

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.

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.

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.

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.

Udder skin surface temperature variation pre- and post- milking in dairy cows as determined by infrared thermography.

The objectives of the research reported in this Research Communication were to compare the variation of hind quarter skin surface temperature pre- and post- milking in dairy cows and to determine the optimal time to capture images by infrared thermography for improving the sensitivity and specificity of mastitis detection in dairy cows. Hind quarter infrared images of 102 Holstein dairy cows were captured from the caudal view by an infrared camera pre-milking and post-milking. The udder skin surface temperature was measured with the help of the image processing software. No significant difference was found between the left and right quarter skin surface temperature pre- and post- milking. The hind quarter skin surface temperature pre-milking was not significantly influenced by milk yield, but exhibited a rising trend along with the increase of milk yield. The hind quarter skin surface temperature post-milking was significantly influenced by milk yield. This leads us to conclude that the sensitivity and specificity of IRT in mastitis detection may be influenced by milk yield and it may be better to capture the infrared images of cow udders pre-milking.

Chronic heat stress increases insulin-like growth factor-1(IGF-1) but does not affect IGF-binding proteins in growing pigs.

The insulin-like activity of insulin-like growth factor-1 (IGF-1) is heavily blunted by IGF binding proteins (IGFBPs, including IGFBP-1 and IGFBP-3). The effects of heat stress (HS) on the IGF-1 and IGFBPs in growing pigs are still not clear. This study aimed to investigate the alterations of IGF-1 and IGFBPs under chronic HS in growing pigs. Twenty-seven growing large white barrows with similar body weight were collected from nine litters and were assigned into three treatments. The litter effect is balanced in all treatments. Treatments were: 1) thermal-neutral (TN) group (23 °C), 2) chronic HS group (33 °C), and 3) pair-fed in TN condition (PFTN). The experiment lasted for 21 days. Compared with TN controls, decreased FI, lower average body weight gain, higher rectal temperature and increased respiration rates were observed in HS pigs. On D7, increased plasma insulin concentration and insulin:glucose ratio were observed in HS pigs compared to TN controls. A overall elevation of plasma IGF-1:IGFBP-3 ratio was detected in HS pigs compared with PFTN and TN counterparts. Besides, hepatic IGF-1 gene expression of HS pigs was 50% higher than TN counterparts. PFTN pigs, however, had no differences in plasma IGF-1:IGFBP-3 ratio and hepatic IGF-1 gene expression, compared with TN pigs. PFTN pigs increased plasma IGFBPs concentration and hepatic IGFBPs gene expression, compared with TN controls. However, no differences in plasma IGFBPs concentration and hepatic IGFBPs gene expression were observed between TN and HS group. Liver IRS-1 gene and protein expressions of HS pigs tended to be increased compared with TN controls, while PFTN pigs still kept liver IRS-1 gene and protein expressions in normal level. High temperature (33 °C), independent of feed intake reduction, increased IGF-1 but did not affect IGFBPs levels in growing pigs.

Proteomic changes of the porcine skeletal muscle in response to chronic heat stress.

BACKGROUND: Heat stress (HS) has an adverse effect on meat quality; however, the underlying molecular mechanisms altering meat quality due to muscle responses to stress remain unclear. Sixteen castrated male crossbreeds between Landrace × Yorkshire sows and Duroc boars (79.00 ± 1.50 kg body weight) were exposed to either thermal neutral (22 °C, n = 8) or HS (30 °C, n = 8) conditions for 3 weeks. Subsequently, the longissimus dorsi (LD) muscle of all pigs was assayed for meat quality parameters and proteome analysis. RESULTS: HS decreased post mortem (24 h) pH and intramuscular fat, changed ultimate L*, a* and b* values and increased drip loss and shear force. Proteome analysis of the LD was conducted by two-dimensional gel electrophoresis and mass spectrometry. A total of 23 differentially expressed proteins were identified, of which three were verified by western blotting analysis. The identified proteins were involved in six types of biological process: carbohydrate metabolism, myofibrillar and cytoskeleton structure, stress response, antioxidant and detoxification, calcium binding and cellular apoptosis. Interestingly, HS induced higher levels of heat shock protein, antioxidants and calcium binding proteins, which are involved in the mechanisms of defense and homeostasis. CONCLUSION: The results indicate that HS-induced changes in the expression of myofibrillar proteins, glucose and energy metabolism-related proteins, heat shock protein and antioxidant enzymes might, at least partly, contribute to increase in meat tenderness. These findings will provide the foundation for developing future mitigating solutions and preventative therapies to reduce the detrimental effects of chronic HS on muscle function, metabolism and meat quality. © 2017 Society of Chemical Industry.

Transcriptome analysis reveals that constant heat stress modifies the metabolism and structure of the porcine longissimus dorsi skeletal muscle.

Exposure to high ambient temperatures is detrimental to pig rearing and porcine meat quality. Deep molecular sequencing allows for genomic characterization of porcine skeletal muscles and helps understand how the genomic landscape may impact meat quality. To this end, we performed mRNA-seq to molecularly dissect the impact of heat stress on porcine skeletal muscles, longissimus dorsi. Sixteen castrated, male DLY pigs [which are crossbreeds between Duroc (D) boars and Landrace (L) × Yorkshire (Y) sows, 79.0 ± 1.5 kg BW] were evenly split into two groups that were subjected to either control (CON) (22 °C; 55 % humidity) or constant heat stress (H30; 30 °C; 55 % humidity) conditions for 21 days. Seventy-eight genes were found to be differentially expressed, of which 37 were up-regulated and 41 were down-regulated owing to constant heat stress. We predicted 5247 unknown genes and 6108 novel transcribed units attributed to alternative splicing (AS) events in the skeletal muscle. Furthermore, 30,761 and 31,360 AS events were observed in the CON and H30 RNA-seq libraries, respectively. The differentially expressed genes in the porcine skeletal muscles were involved in glycolysis, lactate metabolism, lipid metabolism, cellular defense, and stress responses. Additionally, the expression levels of these genes were associated with variations in meat quality between the CON and H30 groups, indicating that heat stress modulated genes crucial to skeletal muscle development and metabolism. Our transcriptomic analysis provides valuable information for understanding the molecular mechanisms governing porcine skeletal muscle development. Such insights may lead to innovative strategies to improve meat quality of pigs under heat stress.

Chronic Heat Stress Induces Immune Response, Oxidative Stress Response, and Apoptosis of Finishing Pig Liver: A Proteomic Approach.

Heat stress (HS) negatively affects human health, animal welfare, and livestock production. We analyzed the hepatic proteomes of finishing pigs subjected to chronic heat stress (HS), thermal neutral (TN), and restricted feed intake conditions, identifying differences between direct and indirect (via reduced feed intake) HS. Twenty-four castrated male pigs were randomly allocated to three treatments for three weeks: (1) thermal neutral (TN) (22 °C) with ad libitum feeding; (2) chronic HS (30 °C) with ad libitum feeding; and (3) TN, pair-fed to HS intake (PF). Hepatic proteome analysis was conducted using two-dimensional gel electrophoresis and mass spectrometry. Both HS and PF significantly reduced liver weight (p < 0.05). Forty-five hepatic proteins were differentially abundant when comparing HS with TN (37), PF with TN (29), and HS with PF (16). These proteins are involved in heat shock response and immune defense, oxidative stress response, cellular apoptosis, metabolism, signal transduction, and cytoskeleton. We also observed increased abundance of proteins and enzymes associated with heat shock response and immune defense, reduced the redox state, enhanced multiple antioxidant abilities, and increased apoptosis in HS liver. Heat-load, independent of reduced feed intake, induced an innate immune response, while food restriction caused stress and cellular apoptosis. Our results provide novel insights into the effects of chronic HS on liver.

Nutritional and physiological responses of finishing pigs exposed to a permanent heat exposure during three weeks.

The aim of the current study was to investigate the effect of a permanent heat exposure during 21 days on pig performance, nutrient digestibility, physiological response and key enzyme of skeletal muscle energy metabolism. Twenty-four male finishing pigs (crossbreed castrates, 79.0 ± 1.50 kg body weight) were allocated to three groups (n = 8): (1) Control (ambient temperature (AT) 22°C, ad libitum feeding), (2) Group HE (AT 30°C, ad libitum feeding) and (3) Group PF (AT 22°C, pair-fed to Group HE). The permanent heat exposure decreased feed intake (p < 0.01), daily body weight gain (p < 0.05) and the digestibility of gross energy, dry matter, crude protein and ash (p < 0.05); rectal temperature and respiration rate were significantly increased (p < 0.01). The levels of plasma cortisol, creatine kinase and lactate dehydrogenase were also significantly increased in Group HE (p < 0.05). Furthermore, the heat exposure changed intracellular energy metabolism, where the AMP-activated protein kinase was activated (p = 0.02). This was combined with changes in parameters of glycolysis such as an accumulation of lactic acid (p = 0.02) and a drop of pH24 h (p = 0.02), an increase of hexokinase and pyruvate kinase activity (p < 0.01) and, finally, the maturation process of post mortem muscle was influenced. Due to pair-feeding it was possible to evaluate the effects of heat exposure, which were not dependent on reduced feed intake. Such effects were, e.g., reduced nutrient digestibility and changed activities of several enzymes in muscle and blood serum.

The Expression of Carnosine and Its Effect on the Antioxidant Capacity of Longissimus dorsi Muscle in Finishing Pigs Exposed to Constant Heat Stress.

The objective of this study was to assess the effects of constant high ambient temperatures on meat quality, antioxidant capacity, and carnosine expression in longissimus dorsi muscle of finishing pigs. Castrated 24 male DLY (crossbreeds between Landrace×Yorkshire sows and Duroc boars) pigs were allocated to one of three treatments: constant ambient temperature at 22°C and ad libitum feeding (CON, n = 8); constant high ambient temperature at 30°C and ad libitum feeding (H30, n = 8); and constant ambient temperature at 22°C and pair-fed with H30 (PF, n = 8). Meat quality, malondialdehyde (MDA) content, antioxidant capacity, carnosine content, and carnosine synthetase (CARNS1) mRNA expression in longissimus dorsi muscle were measured after three weeks. The results revealed that H30 had lower pH24 h, redness at 45 min, and yellowness at 24 h post-mortem (p<0.05), and higher drip loss at 48 h and lightness at 24 h post-mortem (p<0.01). Constant heat stress disrupted the pro-oxidant/antioxidant balance in longissimus dorsi muscle with higher MDA content (p<0.01) and lower antioxidant capacity (p<0.01). Carnosine content and CARNS1 mRNA expression in longissimus dorsi muscle of H30 pigs were significantly decreased (p<0.01) after three weeks at 30°C. In conclusion, constant high ambient temperatures affect meat quality and antioxidant capacity negatively, and the reduction of muscle carnosine content is one of the probable reasons.

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.

Corrigendum: Effects of heat stress on endocrine, thermoregulatory, and lactation capacity in heat-tolerant and -sensitive dry cows.

[This corrects the article DOI: 10.3389/fvets.2024.1405263.].

Effects of heat stress on endocrine, thermoregulatory, and lactation capacity in heat-tolerant and -sensitive dry cows.

Introduction: Increasing global temperatures present a significant challenge to livestock production. The dry period is an important stage in the production cycle of cow, and environmental heat stress (HS) during this period can have adverse effects on the subsequent lactation performance. In this study, we aimed to investigate the effects of HS on endocrine, thermoregulatory, and lactation parameters in heat-tolerant dry cows (HTDC) and heat-sensitive dry cows (HSDC). Methods: We measured the respiratory rate (RR), body temperature (BT), and temperature-humidity index (THI) in 66 dry cows during HS. The slopes of RR and BT to THI were determined through analysis of measurements and dry cows background information using a mixed-effects model. Subsequently, the heat tolerance or sensitivity of dry cows was assessed using clustering method (HTDC = 19 and HSDC = 47). Results: Compared with that of HSDC, the RR of HTDC significantly increased after exposure to HS (p < 0.05). The average reduction in milk yield from new lactation to the previous lactation was significantly lower in HTDC compared to HSDC (p < 0.05). Plasma cortisol and non-esterified fatty acid levels were significantly lower in HTDC compared to HSDC (p < 0.05), while plasma triiodothyronine (p = 0.07) and growth hormone (p = 0.08) levels tended to be higher in HTDC relative to HSDC. Discussion: HTDC can more effectively alleviate the impacts of HS through their superior thermoregulation and metabolism, thereby ensuring optimal postpartum lactation performance.

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.

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.

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.

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.