Mechanism and functional substances of Saiga antelope horn in treating hypertension with liver-yang hyperactivity syndrome explored using network pharmacology and metabolomics.

ETHNOPHARMACOLOGICAL RELEVANCE: Saiga antelope horn (SAH) is a traditional Chinese medicine for treating hypertension with liver-yang hyperactivity syndrome (Gan-Yang-Shang-Kang, GYSK), that has a long history of clinical application and precise efficacy, but its mechanism and functional substances are still unknown. Based on the demand for alternative research on the rare and endangered SAH, the group designed and carried out the following studies. AIM OF THE STUDY: The purpose of this research was to demonstrate the functional substances and mechanisms of SAH in the treatment of GYSK hypertension. MATERIALS AND METHODS: The GYSK-SHR model was constructed by administering a decoction of aconite to spontaneously hypertensive rats (SHRs). Blood pressure (BP), behavioural tests related to GYSK, and pathological changes in the kidneys, heart and aorta were measured to investigate the effects of SAH on GYSK-SHRs. Proteomic analysis was used to identify the keratins and peptides of SAH. Moreover, network pharmacology and plasma metabolomics studies were carried out to reveal the mechanisms by which functional peptides in SAH regulate GYSK-hypertension. RESULTS: SAH has a significant antihypertensive effect on GYSK hypertensive animals. It has also been proven to be effective in protecting the function and structural integrity of the kidneys, heart and aorta. Moreover, SAH improved the abnormalities of 31 plasma biomarkers in rats. By constructing a "biomarker-target-peptide" network, 10 functional peptides and two key targets were screened for antihypertensive effects of SAH. The results indicated that SAH may exert a therapeutic effect by re-establishing the imbalance of renin-angiotensin (RAS) system. CONCLUSIONS: Functional peptides from keratin contained in SAH are the main material basis for the treatment of GYSK-hypertension and exhibited the protective effect on the GYSK-SHR model through the RAS system.

Calves peak-end memory of pain.

In humans, the 'peak-end' rule states that recollection of an experience is most often influenced by the peak (the most intense moment) and end of the experience. We investigated whether calves followed the peak-end rule in their memory of a painful procedure: disbudding. As proxies for retrospective and 'real-time' reports of pain, we used conditioned place aversion, and reflex pain behaviours. In two separate trials, calves were subjected to two disbudding conditioning sessions (one horn per treatment), acting as their own control. In the first trial, calves (n = 22) were disbudded and remained in a pen for 4 h, and disbudded and left in another pen for 4 h with an additional 2 h following an analgesic treatment. In the second trial, calves (n = 22) were disbudded and left in pens for 6 h during both treatments, receiving the analgesic at either 2 h or 4 h after disbudding. Calves were then tested for place aversion. For both trials we did not observe a preference for the pens where calves received analgesic treatment towards the end of the session. We did not find an association between aversion and the sum, peak or end of pain behaviours. Results are not consistent with a peak-end effect in calves' memory of pain.

Combination of mathematics and label-free proteomics for discovering keratin-derived specific peptide biomarkers to distinguish animal horn-derived traditional Chinese medicines.

Saiga antelope horn and Rhinoceros horn have been used in traditional Chinese medicine for thousands of years. However, due to the protection of wildlife, the application of these rare animal horns has been restricted or prohibited. Therefore, water buffalo horn, goat horn, and yak horn have been applied as alternatives to Rhinoceros horn or Saiga antelope horn in a clinic. It is extremely difficult to distinguish normal animal horns in powdered or decocted form, especially identifying related species such as water buffalo horn, yak horn, and cattle horn. In this work, mathematics set and label-free proteomics analysis were combined for discovering keratin-derived specific peptide biomarkers. By using mathematics set analysis after nano liquid chromatography-tandem mass spectrometry-based proteomics, the selected species-specific peptides could be used to identify the authenticity of the Saiga antelope horn and goat horn. Furthermore, peptide biomarkers were selected to distinguish related species-derived horns, water buffalo horn, yak horn, and cattle horn. In total, eight peptide biomarkers were selected and applied for simultaneously distinguishing different horn samples. The present strategy provides a method for peptide biomarkers discovery and also has positive significance for ensuring the quality and efficacy of animal horn-derived traditional Chinese medicines and their products.

Indication of social buffering in disbudded calves.

Most dairy calves are housed individually in early ontogeny but social housing has positive effects on calf welfare including an advantage of social buffering, i.e., when negative effects of stress are mitigated through social support of conspecific. The effects of social buffering has not yet been examined in relation to disbudding; a painful husbandry procedure commonly performed on young dairy calves. The objective of this study was to investigate the effect of pair versus individual housing on calves' behavioral reaction to disbudding. In total 52 female calves were randomly allocated either to individual (n = 16) or pair housing (n = 36, 18 focal). Calves were hot-iron disbudded with a local anesthetic and their spontaneous behavior in home pens was recorded for 24 h pre- and post-disbudding. Eating forage, ruminating, resting, exploration, play, self-grooming, and pain-related behaviors were quantified during eight 20 min intervals during the 24 h periods pre- as well as post-disbudding. In pair-housed (PAIR) calves social resting, active and passive allo-grooming were additionally recorded. The differences between individually housed (INDI, n = 10) and PAIR calves (n = 12) were tested by general linear models. The changes in pre- and post-disbudding behaviors in all calves as well as in social behaviors of PAIR calves were tested by paired t-test. We found that head shaking (t = - 3.46, P = 0.0024), head rubbing (t = 4.96, P < 0.0001) and self-grooming (t = 2.11, P = 0.04) increased in all calves after disbudding. Eating forage increased only in PAIR calves (t = 2.50, P = 0.030) which also resulted in a difference between treatments with PAIR calves fed more often than INDI calves (F1,18 = 12.96, P = 0.002). Differences in eating forage may be an indication of improved ability of PAIR calves to recover from disbudding. No other significant differences were detected between treatment groups which might have been caused by our limited sample. Our results provide the first evidence that housing treatment affects calves' reactions to disbudding, with possible indication of social buffering.

Effect of plane of nutrition and analgesic drug treatment on wound healing and pain following cautery disbudding in preweaning dairy calves.

The objective of this study was to determine the effect of a biologically normal plane of nutrition compared with a limited plane on the primary outcome wound healing, and one dose of nonsteroidal anti-inflammatory drug (NSAID) compared with 2 on the secondary outcomes: lying behavior, haptoglobin concentrations, and mechanical nociceptive threshold (MNT) in calves disbudded via cautery iron. Eighty female Holstein calves were enrolled at birth, individually housed, and fed via a Calf Rail system (Förster Technik). A 2 × 2 factorial design was used to assess the effect of plane of nutrition and an additional NSAID. Calves were randomly assigned to a biologically normal plane of nutrition (BN; offered up to 15 L/d) or a limited plane (LP; offered up to 6 L/d) and to receive one or 2 doses of meloxicam. All calves received a lidocaine cornual nerve block and a subcutaneous injection of meloxicam 15 min before cautery disbudding at 18 to 25 d of age, and half the calves received an additional injection of meloxicam (0.5 mg/kg) 3 d after disbudding. Tissue type present, wound diameter, and wound depth were evaluated 2 times per week for 7 to 8 wk as measures of wound healing, lying behavior was recorded beginning 1 to 2 wk before disbudding until 7 to 8 wk after as a behavioral indicator of pain, haptoglobin concentrations were measured once per day for 7 d after disbudding, and MNT was evaluated 2 times/wk for 3 wk. Survival analyses were analyzed using Cox regression models (wound healing) and continuous data were analyzed using mixed-effect linear regression models. Only 12% of horn buds were completely healed by 7 to 8 wk after disbudding and 54% had re-epithelized at this time. At any time, wounds from BN calves were more likely to have had re-epithelization occur compared with wounds from LP calves (hazard ratio: 1.93, 95% CI: 1.18-3.14). Wounds from calves that received only one dose of NSAID were more likely to have re-epithelization occur, compared with wounds from calves given 2 doses (hazard ratio: 1.87, 95% CI: 1.15-3.05). Wounds from BN calves had smaller diameters and depths over time beginning on wk 3 compared with LP calves. Wounds from calves that received an additional NSAID had larger diameters and depths over time beginning on wk 4 and 3 respectively, compared with calves that only received one dose of NSAID. Calves that received an extra NSAID tended to be less sensitive 7, 10, and 17 d after disbudding compared with calves that only received one dose and spent less time lying in the week after disbudding. Calves on the BN milk program were more active compared with LP calves with lower lying times, fewer lying bouts per day, and longer average lying bouts. Our results indicate that a BN milk feeding program for calves can result in faster healing times and more activity, and that providing an extra NSAID 3 d after disbudding appears to slow the healing process but may result in less pain experienced by the calf 1 to 2 wk after the procedure. This study is also among the first to demonstrate that after the complete removal of the horn bud, wounds can take more than 8 weeks to re-epithelize and fully heal.

Buffering lidocaine heightens aversion to cornual nerve injections in dairy calves.

Best practice is to numb the horn buds before disbudding dairy calves, which can be achieved by injecting lidocaine, a local anesthetic, around the cornual nerve. In humans, the acute pain that occurs upon injection of lidocaine can be reduced by neutralizing the pH of the solution with an alkalizing agent, such as sodium bicarbonate. We evaluated whether buffering lidocaine would reduce calves' behavioral and physiological responses to cornual nerve blocks on the left and right sides of the head. Female Holstein calves were assigned to 1 of 3 treatments: cornual nerve blocks with 5.5 mL of unbuffered lidocaine (n = 9), buffered lidocaine (n = 9), or a sham procedure with no needle inserted (n = 9). Calves that received either type of lidocaine struggled more during the injection than sham calves. However, contrary to our hypothesis, struggling was most marked in calves that received buffered lidocaine. Similarly, calves administered unbuffered or buffered lidocaine had elevated heart rates for 1 or 3 min after the first injection, respectively, compared with at the end of the 5-min observation period. Calves in the buffered treatment had lower eye temperatures in the first half of the observation period compared with the second half, consistent with responses cattle show to other aversive procedures, but no changes over time were observed in the other 2 treatments. We detected no treatment differences in heart rate variability measures. These results suggest that cornual nerve blocks are aversive, at least in the short term, and that buffering the lidocaine worsens the calf's response to this procedure.

[Identification of specific peptides from Bubali Cornu by ultra-performance liquid chromatography-tandem mass spectrometry].

This paper explored the specific peptides from Bubali Cornu by ultra-performance liquid chromatography-tandem mass spectrometry and based on mathematics set theory. Following the profile analysis of peptides from Bubali Cornu, Bovis Grunniens Cornu, Caprae Hircus Cornu, and Suis Cornu by nano LC-LTQ-Obitrap-MS after digestion with trypsin, the relationship of peptide composition among different samples was analyzed using the mathematics set theory. The ones that existed only in the Bubali Cornu set rather than in any other set were considered as the specific peptides of Bubali Cornu. The further bioinformatic analysis revealed four specific peptides from Bubali Cornu, whose specificity was verified by UPLC-QQQ-MS. The results showed that these four peptides could be used for distinguishing Bubali Cornu from Caprae Hircus Cornu and Suis Cornu. This study has provided a rapid and simple method for seeking the specific peptides in animal medicines, which can be utilized for quality evaluation of animal medicines, thus making them authenticable and traceable.

The effect of transdermal flunixin meglumine on blood cortisol levels in dairy calves after cautery disbudding with local anesthesia.

The objective of this study was to evaluate the effect of the nonsteroidal anti-inflammatory drug transdermal flunixin meglumine (Finadyne Transdermal) on plasma cortisol, average daily weight gain, and standing and lying behavior of calves, when given at the time of disbudding combined with local anesthesia. A sedative was not used to minimize pharmacological interactions. Seventy-one female Holstein Friesian calves aged 13 ± 2 d, with an average weight of 48.9 ± 4.26 kg were enrolled in the study. All calves were randomly assigned to one of 3 treatment groups: (1) control group (CON, n = 27), (2) 1-flunixin group (1-FLU, n = 26) with a single administration of transdermal flunixin meglumine at disbudding, and (3) 2-flunixin group (2-FLU, n = 24) with 2 administrations of transdermal flunixin meglumine, the first treatment at disbudding and the second 6 h after disbudding. Although the CON group received a placebo, 1-FLU and 2-FLU received flunixin meglumine transdermally. To account for plasma cortisol changes due to manipulation and handling of the calves, a sham disbudding procedure was performed one week before disbudding took place. Sham disbudding was conducted by using a cold cautery dehorner applied to each horn bud for 10 s. Disbudding was performed in a similar way by using a hot cautery dehorner. Plasma samples were collected to measure the stress biomarker cortisol at 7 different time points. Body weights were measured 4 times in 2 wk. Standing and lying behavior was assessed via 3-dimensional accelerometer. During sham disbudding and disbudding mean plasma cortisol concentrations were 6.09 ± 2.5 ng/mL and 5.16 ± 2.8 ng/mL, respectively. Treatment tended to have an effect on plasma cortisol concentrations during sham disbudding and had an effect on plasma cortisol concentrations during disbudding. Plasma cortisol concentrations were affected by treatment 2 h after disbudding in comparison to CON group. Furthermore, there was a significant effect on plasma cortisol concentrations 6 h after disbudding in contrast to CON. A return to baseline plasma cortisol levels (initial concentrations) was not achieved in CON during disbudding. There was no statistical difference between average daily weight gain and the treatment procedure. Total lying time was not affected by treatment after disbudding. In conclusion, transdermal flunixin meglumine given at the time of disbudding combined with local anesthesia decreased concentrations of the stress biomarker cortisol, but a second dose 6 h after disbudding had no further effect on plasma cortisol levels.

A comparison of sedative effects of xylazine alone or combined with levomethadone or ketamine in calves prior to disbudding.

OBJECTIVE: To compare the sedative effects of intramuscular xylazine alone or combined with levomethadone or ketamine in calves before cautery disbudding. STUDY DESIGN: Randomized, blinded, clinical trial. ANIMALS: A total of 28 dairy calves, aged 21 ± 5 days and weighing 61.0 ± 9.3 kg (mean ± standard deviation). METHODS: Calves were randomly allocated to three groups: xylazine (0.1 mg kg-1) and levomethadone (0.05 mg kg-1; group XL), xylazine (0.1 mg kg-1) and ketamine (1 mg kg-1; group XK) and xylazine alone (0.2 mg kg-1; group X). Local anaesthesia (procaine hydrochloride) and meloxicam were administered subcutaneously 15 minutes after sedation and 15 minutes before disbudding. The calves' responses to the administration of local anaesthesia and disbudding were recorded. Sedation was assessed at baseline and at intervals up to 240 minutes postsedation. Times of recumbency, first head lift and first standing were recorded. Drug plasma concentrations were measured. RESULTS: Data were obtained from 27 animals. All protocols resulted in sedation sufficient to administer local anaesthesia and to perform disbudding. Sedation scores significantly correlated with drug plasma concentrations (p ≤ 0.002). Times to recumbency did not differ among protocols (2.8 ± 0.3, 3.1 ± 1.1 and 2.1 ± 0.8 minutes for groups XL, XK and X, respectively), whereas interval from drug(s) administration until first head lift was significantly shorter in group XK than X (47.3 ± 14.1, 34.4 ± 5.3 and 62.6 ± 31.9 minutes for groups XL, XK and X, respectively). The area under the time-sedation curve was significantly greater in group X than XK or XL (754 ± 215, 665 ± 118 and 1005 ± 258 minutes for groups XL, XK and X, respectively). CONCLUSIONS AND CLINICAL RELEVANCE: Levomethadone or ketamine with a low dose of xylazine produced short but sufficient sedation for local anaesthesia and disbudding with minimum resistance.

Dairy calves show behavioural responses to hot iron disbudding after local anaesthesia with procaine.

BACKGROUND: The efficacy of procaine as anaesthetic for disbudding of calves is largely unknown. Likewise, information about effect of interval from local anaesthesia to disbudding and the size of the hot iron used is missing. METHODS: A total of 167 dairy calves were disbudded after cornual nerve block with procaine. Time from administration of anaesthetic to disbudding varied, and calves were disbudded using two different sizes of hot iron. The calves' behavioural responses to disbudding were recorded based on (1) getting up, (2) kicking, and (3) lifting head. Using logistic regression, the associations between calves showing at least one of these three behaviours and interval from administration of anaesthetic to disbudding and size of the hot iron were evaluated. RESULTS: Overall, 42% of calves showed at least one of the three behaviours, even though they had received a cornual nerve block prior to disbudding. Using the large hot iron was associated with a 2.3 times higher odds of the calf reacting behaviourally compared to the small iron. Interval from administration of anaesthetic to disbudding was not significantly associated with the odds of behavioural responses. CONCLUSION: Approximately 40% of calves disbudded after cornual nerve block with procaine showed signs of inadequate local anaesthesia. This was especially problematic when hot irons with larger tips were used.

Conditioned place aversion of caustic paste and hot-iron disbudding in dairy calves.

Cauterization by hot iron and application of caustic paste are 2 common methods of disbudding calves. In this study, we compared the affective experience of these 2 procedures on young dairy calves using conditioned place aversion. Male dairy calves (n = 14; 7 ± 2 d old) were disbudded by both thermal and chemical methods (1 horn bud at a time, 48 h apart). Calves received treatments in pens made visually distinct with either red squares or blue triangles on the walls. Calves were restricted to these treatment pens for 6 h following disbudding. For all treatments, calves received a sedative (xylazine, 0.2 mg/kg), local anesthetic (lidocaine, 5 mL), and analgesic (meloxicam, 0.5 mg/kg). Calves were then tested for conditioned place aversion at 48, 72, and 96 h after their last treatment. During tests, calves were placed in a neutral pen connected to both treatment pens where they had previously been disbudded. Time spent in each treatment pen was recorded until calves chose to lie down for 1 min (latency to lie down: 31.0 ± 8.6 min). During the first test (48 h after last disbudding), calves spent more time in the pen associated with hot-iron disbudding compared with what would be expected by chance (intercept: 73.5%, 95% CI: 56.5, 90.5) and fewer calves lay down in the caustic paste pen than in the hot-iron pen (3 vs. 10 lying events). No evidence of preference for the hot-iron pen was found in the following test sessions (72 and 96 h since last disbudding). These results suggest that calves initially remember caustic paste disbudding as a more negative experience than hot-iron disbudding, even with the use of sedation, local anesthesia, and analgesia.

Behavioral changes in calves 11 days after cautery disbudding: Effect of local anesthesia.

Hot-iron disbudding results in painful burn wounds that take weeks to heal. Spontaneous behaviors indicative of pain are apparent in the immediate hours after disbudding, but whether they occur later in the healing process is unknown. To evaluate whether ongoing pain was present around the time the necrotic tissue loosens from the scalp, we tested the effect of administration of local anesthetic 11 d after the procedure. Disbudded female Holstein and Jersey calves (n = 24) were randomly assigned to receive an injection of local anesthetic (lidocaine) or saline at the cornual nerve on both sides of the head. We recorded the frequency of 8 behaviors for 75 min: head shakes, head rubs, head scratches, ear flicks, tail flicks, bucks/jumps/kicks, grooming, and transitions between standing and lying. Calves treated with lidocaine shook their heads less and tended to flick their ears less than calves administered saline, consistent with the effects of pain relief previously reported in the immediate hours after disbudding. These calves also rubbed their head against the sides of the pen more often, suggesting lidocaine suppressed wound protective behavior. Head shaking and head scratching became more common in the last 25 min compared with the first 50 min in calves treated with lidocaine, consistent with the return of sensation to the disbudding wounds. No treatment differences in the other behaviors were observed. These results suggest that calves experience ongoing pain 11 d after hot-iron disbudding, adding to a growing body of evidence that pain persists for weeks after the procedure.

Randomized control trial assessing the efficacy of pain control strategies for caustic paste disbudding in dairy calves younger than 9 days of age.

Disbudding is a common procedure practiced in the dairy industry and is known to cause pain when performed without pain control. Dairy producers who disbud calves with caustic paste are less likely to provide pain control than those using cautery. Little research has been conducted on pain control for caustic paste disbudding and no studies have specifically examined calves under 9 d of age. The objective of this study was to evaluate the efficacy of local anesthesia and nonsteroidal anti-inflammatory drug analgesia on indicators of pain and inflammation in dairy calves disbudded using caustic paste. One hundred forty Holstein heifer calves 1 to 9 d of age were enrolled in 28 blocks and randomly allocated to 1 of 5 treatments: sham control (SH); positive control (POS); lidocaine cornual nerve block (LC); meloxicam (MEL); and lidocaine cornual nerve block plus meloxicam (LCM). We measured outcomes including serum cortisol and haptoglobin, pressure sensitivity, and lying behavior. Data were analyzed using mixed linear regression models with treatment as a fixed effect, baseline values as a covariate, and trial block as a random effect. Compared with the POS group, the LCM group had reduced serum cortisol at 15, 30, 45, and 60 min post-disbudding; cortisol values were not different between LC, LCM, and SH calves at these time points. At 60, 90, 120, and 180 min post-disbudding, LCM calves had reduced cortisol compared with LC calves, whereas, values did not differ between LCM and SH calves at these time points. At 3 to 4 d post-disbudding, the LCM group tended to have reduced haptoglobin, but no differences were found between groups at 180 min and 7 d post-disbudding. At 60, 90, and 120 min post-disbudding, LC and LCM treated calves had decreased pressure sensitivity compared with other groups. No differences were seen in pressure sensitivity between groups at 180 min, 3 to 4 or 7 d post-disbudding. No differences in lying behavior were found between treatment groups on any of the 7 d following disbudding. These findings demonstrate that the combination of a local anesthetic with a nonsteroidal anti-inflammatory drug is beneficial for reducing indicators of pain and inflammation in young calves disbudded with caustic paste.

Acute cortisol and behavior of dairy goat kids administered local anesthesia, topical anesthesia or systemic analgesia prior to cautery disbudding.

Cautery disbudding of goat kids causes thermal burns and tissue destruction, which results in acute and post-operative pain and negatively affects animal welfare. The objectives of this study were to evaluate acute cortisol concentrations and behavioral responses associated with (1) injecting a lidocaine ring block prior to cautery disbudding and comparing this to saline injections and (2) pain mitigation strategies (lidocaine ring block, topical eutectic mixture of local anesthetics (EMLA) cream, oral meloxicam) on cautery disbudded dairy goat kids. Sixty doe kids were allocated to one of six treatments: (1) disbudding without pain relief (DB), (2) a ring block using 1% lidocaine (DBLA) 20 min before disbudding, (3) saline injection (DBSA) 20 min before disbudding, (4) oral meloxicam 60 min before disbudding (DBMEL), (5) EMLA cream rubbed into the buds 60 min before disbudding (DBEM) and (6) handled but not disbudded (HAND). Blood was sampled pre- (-20 and -5 min) and post-treatment (15 and 30 min) to assess plasma cortisol concentrations and behavior during treatment was recorded using video cameras to assess rump movements, tail shakes and vocalizations (values presented as number/treatment). DBLA and DBSA kids showed elevated cortisol above baseline 5 min pre-disbudding (after injections) (P ≤ 0.01), which was no different to cortisol 15 min post-disbudding (P > 0.05). Rump movements and tail shakes of DBLA (5.5 ± 0.8 and 6.9 ± 1.2) and DBSA kids (5.6 ± 0.8 and 7.2 ± 1.2) were no different to those of DB kids (P > 0.10). Cortisol was elevated from baseline for 30 min post-disbudding for DBEM kids and DBMEL kids (P ≤ 0.05). Rump movements, tail shakes and vocalizations of DBEM (5.7 ± 0.8, 6.3 ± 1.2 and 11.1 ± 1.6) and DBMEL kids (5.3 ± 0.8, 8.0 ± 1.2 and 9.1 ± 1.6) were no different to those of DB kids (P > 0.05). HAND kids showed no change in cortisol over time (P > 0.10) and performed fewer rump movements, tail shakes and vocalizations than all treatments (P ≤ 0.01). In conclusion, lidocaine injection using a ring block appears to cause more pain than handling alone and may not reduce pain associated with cautery disbudding; therefore, a lidocaine ring block may not be a useful pain mitigation strategy for disbudding dairy goat kids. There was no evidence that meloxicam reduced acute pain and EMLA cream may have intensified the pain associated with disbudding. Further research on efficacious pain mitigation strategies for cautery disbudding of dairy goat kids is required.

Development of an on-line Raman spectral analytical method for monitoring and endpoint determination of the Cornu Caprae Hircus hydrolysis process.

OBJECTIVES: Cornu Caprae Hircus (goat horn, GH), a medicinal animal horn, is frequently used in traditional Chinese medicine, and hydrolysis is one of the most important processes for GH pretreatment in pharmaceutical manufacturing. In this study, on-line Raman spectroscopy was applied to monitor the GH hydrolysis process by the development of partial least squares (PLS) calibration models for different groups of amino acids. METHODS: Three steps were considered in model development. In the first step, design of experiments (DOE)-based preprocessing method selection was conducted. In the second step, the optimal spectral co-addition number was determined. In the third step, sample selection or reconstruction methods based on hierarchical clustering analysis (HCA) were used to extract or reconstruct representative calibration sets from the pool of hydrolysis process samples and investigated for their ability to improve model performance. KEY FINDINGS: This study has shown the feasibility of using on-line Raman spectral analysis for monitoring the GH hydrolysis process based on the designed measurement system and appropriate model development steps. CONCLUSIONS: The proposed Raman-based calibration models are expected to be used in GH hydrolysis process monitoring, leading to more rapid material information acquisition, deeper process understanding, more accurate endpoint determination and thus better product quality consistency.

Assessing the affective component of pain, and the efficacy of pain control, using conditioned place aversion in calves.

Pain in animals is typically assessed using reflexive and physiological responses. These measures allow inferences regarding nociception but provide little basis for conclusions about the affective component of pain (i.e. how negatively the experience is perceived). Calves routinely undergo painful procedures on commercial farms, including hot-iron disbudding, providing a convenient model to study pain in animals. The aim of this study was to investigate the affective component of post-procedural pain due to hot-iron disbudding, using conditioned place aversion. Calves (n = 31) were subjected to two procedures (one bud at a time): one without post-procedural pain control and the other with the use of a nonsteroidal anti-inflammatory drug (either meloxicam (n = 16) or ketoprofen (n = 15)). All procedures included the use of local anaesthesia (lidocaine). Place conditioning was tested 2 days after the last treatment by allowing calves to freely roam between the pens where they had previously been disbudded. Calves spent more time, and lay down more frequently, in the pen where they received meloxicam compared with the pen where they only received a local block. Surprisingly, calves avoided the pen where they received ketoprofen compared with the control treatment pen. We hypothesize that the shorter duration of action of ketoprofen resulted in increasing pain at the end of the conditioning period, explaining the increased aversion to this treatment. These results illustrate the value of place conditioning paradigms to assess the affective component of pain in animals, and suggest that the animal's evaluation of painful events depends upon the time course of when the pain is experienced.

Effects of a topically applied anaesthetic on the behaviour, pain sensitivity and weight gain of dairy calves following thermocautery disbudding with a local anaesthetic.

Aims: To compare the effect of a topically applied anaesthetic to no pain relief or meloxicam on the behavioural responses, pain sensitivity and weight gain of calves following disbudding with or without sedation. Methods: A total of 364, 2-6 week-old calves from three commercial farms were systematically allocated to one of six treatment groups. All calves received a cornual nerve block prior to disbudding, with half restrained in a crate and half sedated with xylazine. Within these groups one third received no further treatment (control), one third were treated with meloxicam >10 minutes prior to disbudding and one third received a topical anaesthetic applied to the horn bud wounds following disbudding. The frequency of ear flicks, head shakes, head scratches and pain sensitivity of the wound were recorded on up to eight occasions over 24 hours after disbudding. Calves were weighed before, and 7 and 28 days after, disbudding to determine average daily weight gain (ADG). Results: Compared to calves in the crate-control group, all other groups had reduced ear flicks at all times following disbudding (p < 0.01). Treatment with meloxicam and topical anaesthesia in addition to sedation reduced head scratches compared to calves in the crate-control group (p ≤ 0.013). At 22 hours after disbudding head shakes were reduced in sedated calves treated with topical anaesthetic compared to calves in the crate-control group (p < 0.001). Pain sensitivity was lower in all sedated calves than unsedated calves (p < 0.001). The ADG between Days 0-7 was 0.14 (95% CI = 0.015-0.274) kg/day greater in sedated calves treated with meloxicam than calves in the crate-control group (p = 0.03), and the ADG between Days 0-28 tended to be 0.06 (95% CI=-0.01-0.13) kg/day greater in sedated calves treated with topical anaesthetic than calves in the crate-control group (p = 0.09). Conclusion and clinical relevance: Sedation of calves for disbudding reduced the pain experienced in the following 24 hours. There was a benefit to providing calves with topical anaesthetic following disbudding on behavioural responses and pain sensitivity, which was similar to that of treating calves with meloxicam.

[Determination of free thiols in animal horn derived traditional Chinese medicines].

This present study is to detect the content of free thiols(-SH) in the horn derived traditional Chinese medicines( TCMs) from different animals and different regions by using fluorescence derivatization method. TCEP was used as a disulfide bond reducing agent,while SBD-F as a derivatization reagent. Fluorescent spectrophotometry was used to determine the content of-SH,and the maximum excitation wavelength and emission wavelength were set as 375 and 510 nm,respectively. As a result,under the optimized condition,the extraction of Caprae Hircus Cornu showed the highest free-SH concentration,followed by Bovis Grunniens Cornu,Bubali Cornu,and Elaphuri Davidiani Cornu. In the present study,we point out that the-SH-contained components might be the most important material basis in animal horn derived TCMs. With good accurate,sensitive and rapid properties,the present method can provide reference basis for the quality evaluation of animal horn derived TCMs and guides for the investigation on effective material basis.

Management of preweaned bull calves on dairy operations in the United States.

Many calves born on dairy farms are male, but little research to date has focused on the management of these calves. The objective of this study was to survey management practices of preweaned dairy bull calves and compare these practices with those used for heifer calves on the same operations. This study was conducted as part of the USDA National Animal Health Monitoring System Dairy 2014 study and included a sample of 42 operations from 10 states. On average, 2.4% of bull calves died before leaving the operation, with most operations selling bull calves before weaning (97.6%). Bull calves left the operation at an average age of 7.6 d (median = 4 d). Producers reported that 96.3% of bull calves received colostrum. Of bulls that were fed colostrum, the vast majority (95.4%) were hand-fed; the remaining calves received colostrum via a combination of hand-feeding and suckling (3.1%) or by suckling only (1.5%). In contrast, all heifer calves on those operations were only hand-fed colostrum. On average, bull calves received 3.1 L of colostrum at their first feeding at 4.3 h after birth compared with 2.9 L at 2.9 h after birth for heifer calves. In the first 24 h, bull and heifer calves received 4.8 and 5.5 L of colostrum, respectively. Only 21.4% of operations dehorned bull calves; in comparison, heifer calves were dehorned on 90.5% of operations. Few operations used analgesics or anesthetics when dehorning bull calves, whereas 28.9% of operations used anesthetics or analgesics when dehorning heifer calves. Overall, 27.8% of operations castrated bull calves. Of the operations that did castrate bulls, 70.0% used a band at an average age of 6.3 wk; the remaining 30% of operations used a knife at 14.7 wk of age on average. Few operations used analgesics or anesthetics when castrating. These results illustrate differences in the care of bull and heifer calves and suggest the need to improve education regarding recommended colostrum feeding and pain management practices in all calves regardless of sex.

A new method of administering local anesthesia for calf disbudding: Findings from a comparative on-farm study in New Zealand.

Anesthesia of the horn bud for calf disbudding commonly is attained by injection of local anesthetic over branches of the cornual nerve, with anesthesia achieved in 3 to 20 min. With experienced and trained staff, this method is effective in 88 to 100% of calves. Variability in response and time of onset can compromise calf welfare if calves are disbudded before anesthesia is attained. Proposed legislative reliance on effective local anesthetic as the minimal method of pain relief for calves at disbudding means that administration of local anesthetic must achieve a repeatable level and rapid onset of analgesia. We describe an alternative method of local anesthesia administration that uses local site infiltration of anesthetic over the horn bud. However, this method has not yet been scientifically validated. This study assessed differences between disbudding using the cornual nerve block and disbudding with local anesthesia administered by local site infiltration. Efficacy of local anesthesia was assessed at 30-s intervals after administration by absence of reaction to 3 consecutive needle pricks over the horn buds. Behavior indicating pain was assessed during disbudding and scored from 0 to 3. Calf behavior was also recorded for 3 h after disbudding. Accelerometer data loggers were fitted to each calf for 24 h before and after disbudding to assess lying and standing times. Median time to cutaneous desensitization for local infiltration was 60 s compared with 225 s for cornual nerve block, and the variance in time to desensitization was less with local infiltration. Calves disbudded under cornual block had a larger behavioral response (indicated by a graded aversive body reaction) than calves disbudded under local infiltration. A multivariable model predicted that the mean body reaction score would be 0.6 for calves disbudded under local infiltration and 1.2 for calves disbudded under cornual block. There was no difference in any behaviors between the treatment groups in the 3 h after disbudding. Method of analgesia had no effect on lying time over the 24 h after disbudding. In this study, local infiltration was at least as effective in providing analgesia for disbudding as the cornual nerve block. Our results suggest that a more consistent, effective level of analgesia during disbudding was achieved using local infiltration and that there was no difference in postoperative expressions of pain.