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Full Research Article

Evaluation of Clinical and Physiological Parameters Following Ketamine Anaesthesia in Conjunction with Various Premedicants in Male Water Buffalo Calves (Bubalus bubalis)

K
Khichar Sangram Singh1
R
Rukmani Dewangan1,*
https://orcid.org/0000-0002-5826-2391
R
Raju Sharda1
J
Jasmeet Singh1
I
Ishant Kumar1
M
Muskan Sengar1
L
Likchavi Kurrey1
1Department of Veterinary Surgery and Radiology, College of Veterinary Science and A.H., Dau Shri Vasudev Chandrakar kamdhenu Vishwavidyalaya, Anjora, Durg-491 001, Chhattisgarh, India.

ABSTRACT

Background: Ruminants are poor subject for general anaesthesia. With advancement in veterinary sciences, there is need of balanced anaesthesia with rapid, smooth, induction and lesser recumbency time for large animals. Therefore, the present study was planned to evaluate the alternations on clinico-physiological parameters following ketamine anaesthesia in buffalo calves premedicated with either diazepam, butorphanol or xylazine.

Methods: The study was undertaken in eighteen male buffalo calves and divided into three groups (A, B and C) with six animals in each. Ten minutes prior to the anaesthetic administration, all the buffalo calves were administrated glycopyrrolate 0.01 mg kg b.wt. intramuscularly. The animals of group A, B and C were premedicated intravenously with diazepam 0.5 mg kg b.wt., butorphanol 0.075 mg kg b.wt. and xylazine 0.16 mg kg b.wt. respectively. General anaesthesia was induced with ketamine 4 mg kg b.wt. intravenously and clinical parameters were recorded viz., onset of sedation, induction, duration and recovery of anaesthesia. Physiological parameters were also recorded before (0), 5 min after sedation, induction and at 10, 20, 40, 60 and 120 min following ketamine anaesthesia.

Result: The onset of sedation and anaesthesia was quicker in animals of group A followed by group C and B respectively. The physiological parameters showed transient changes in all the three groups which were compensated and remained within normal physiological range during the study period. Ketamine in conjunction with various preanaesthetic combinations did not produce any adverse effect on cardiopulmonary system. Thus, ketamine could be safely used as general anaesthestic in buffalo calves premedicated with diazepam, butorphanol or xylazine. However, a combination of xylazine-ketamine produced longer duration of surgical anaesthesia in buffalo calves as compared to other anaesthetic combinations.

INTRODUCTION

Ruminants are crucial in India for livelihood and poverty reduction, especially for small and landless farmers who can raise them on marginal lands. Buffalo (Bubalus bubalis) is established as the ‘Black gold’ of India because of most valuable species with excellent zootechnical characteristics for both milk and meat production. However, dairy buffalo rearing can only be rewarding if the buffalo calves are healthy and face less mortality during their early life (Siddhapara et al., 2022). Historically, general anaesthesia was considered unsuitable for ruminants due to various challenges and concerns. Its use was primarily restricted to dogs and horses. Buffaloes are generally considered to be poor subjects to general anaesthesia because of tympany, regurgitation, delayed recovery and haemodynamic disturbances in dorsal and lateral recumbency. There is need for an ideal anaesthetic agent in order to produce desired analgesia, sedation, relaxation and safety for body’s vital system which should also be economical and easy to apply (Pemayun and Sudisma, 2020). Moreover, rapid onset along with quick recovery is desirable in large ruminants due to high risk of regurgitation and tympany. Until now, despite the availability of newer anaesthetic and analgesic drugs, there is no anaesthetic agent that meets the ideal anaesthetic requirement. Therefore, suitable drug combinations are needed to produce a state of balanced anaesthesia. Glycopyrrolate is anti-cholinergic drug which is quaternary ammonium salt and can be administered @ 0.01 mg/kg IM in buffalo calves that blocks cardiac vagus and thereby inhibit cardiac inhibitory effects along with alternation in haemodynamic parameters induced by xylazine in buffalo calves (Potliya et al., 2015). Diazepam is a benzodiazepine derivative and possesses anxiolytic, anticonvulsant, hypnotic, sedative, skeletal muscle relaxant and amnestic properties (Ragab et al., 2022). Butorphanol tartrate is an opioid agonist- antagonist with good analgesic, antitussive and sedative properties which is known to induce only mild sedation and has minimum adverse effects to cardiovascular system in buffalo calves (Bodh et al., 2015). Alpha 2 agonists like xylazine are potent sedatives, widely used in both large and small animals including rapid onset, reliable and potent effect, reversibility and analgesia (Mckelvey and Hollingshead, 2003). Ketamine is a non-competitive N-methyl-d-aspartate receptor antagonist which provides sedation, amnesia with analgesia and has anticonvulsive and neuroprotective properties (Sengar et al., 2020). Ketamine alone produces poor muscle relaxation alongwith muscle spasms. To overcome these undesirable effects, ketamine is often combined with alpha 2  adrenoreceptor agonists like xylazine (Eze et al., 2004), opioids like butorphanol (Bodh et al., 2015; Maidanskaia et al., 2023) and benzodizopines like diazepam (Pawde et al., 2000; Nain et al., 2010) which nullify the potential hypertensive caused by ketamine and providing muscle relaxation. A comprehensive and planned study on the use of ketamine hydrochloride effect in conjunction with various preanaesthetic in buffalo calves are scanty and determining the appropriate general anaesthesia protocol for buffalo due its specific physiological characteristic. Hence, the present study was designed to evaluate the clinico-physiological alternations following ketamine anaesthesia in male buffalo calves premedicated with either diazepam, butorphanol or xylazine.

MATERIALS AND METHODS

Anaesthetic protocol
 
The study was conducted on eighteen healthy male buffalo calves having body weight 80-100 kg and randomly divided into three groups A, B and C comprising six animals in each. The buffalo calves were dewormed with Tab Fenbendazole 150 mg-5 mg/kg body weight orally one month prior to the start of anaesthetic study. Ten minutes prior to the anaesthetic administration, all the buffalo calves were administrated glycopyrrolate 0.01 mg kg b.wt. intramuscularly. The animals of group A, B and C were premedicated intravenously with diazepam 0.5 mg kg b.wt., butorphanol 0.075 mg kg b.wt. and xylazine 0.16 mg kg b.wt. respectively. General anaesthesia was induced with ketamine 4 mg kg b.wt. intravenously and clinical parameters recorded were onset of sedation,  induction, duration and recovery from anaesthesia as per Kumar et al., (2014 a,b) and Bodh et al., (2015). Physiological parameters viz. rectal temperature, heart rate and respiratory rate were recorded at 0 (base value), 5 min after sedation and at 10, 20, 40, 60, 120 min. intervals following ketamine anaesthesia.
 
Statistical analysis
 
The data collected were statistically analyzed using Analysis of variance (ANOVA) and Duncan’s Multiple range tests (DMRT) by SPSS v20 statistic software program and all the data were expressed as mean±Standard Error. 

RESULTS AND DISCUSSION

Clinical parameters
 
Onset of sedation
 
Comparison between groups revealed rapid onset of sedation in animals of Group A than Group C and B. Onset of sedation was significantly (P<0.05) shorter in Group A as compared to Group C and B as shown in Table 1. The shorter period of sedation onset was achieved by administration of diazepam which collaborates with the findings of Kumar et al., (2006) and Nain et al., (2010) in buffalo calves and might be due to depression of CNS that is mediated through depression of limbic system and due to enhancement of gamma-amino butyric acid (GABA) by binding of diazepam to GABAA receptor, an inhibitory channel which when activated, decreases neuronal activity (Hall et al., 2001). In Group C, there was decrease in the behavioural activity of animals after administration of xylazine and thereafter animals went to lateral recumbency within 4.11±1.06 min. Similarly, Nirmale et al., (2024) documented average sedation time 3.75±0.25 min.after administration of xylazine intravenously. Onset of sedation was quicker in this group which could be due to sedative activity of α 2 - adrenoceptor agonists in the locus coeruleus, where noradrenergic neurons are found in high concentration (Singh et al., 2006). These findings are concurrent to Habib et al., (2002) in sheep after intramuscular administration of xylazine and Tunio et al., (2003) in goats after intravenous administration of detomidine. Similarly, Kumar et al., (2006) also reported decrease in spontaneous activity where animals went into sternal recumbency by 8.00±1.00 min following administration of detomidine in buffalo calves. Sedative effects of α-2 agonists are associated with the activation of alpha-2 adrenoceptors, which cause decrease in the release and turnover of norepinephrine in the CNS (Shah et al., 2025). In present study, the administration of butorphanol did not affect degree of sedation and temperament of the animal, as a result longer onset of sedation was noticed in animals of group B. Similar finding was also reported by Almubarak (2013) in camels after administration of butorphanol.

Table 1: Showing onset of sedation, induction, duration and complete recovery from anaesthesia in different groups.


 
Induction of anaesthesia
 
Onset of induction was rapid in animals premedicated with diazepam and xylazine as compared to group B (Table 1), which might be due to synergistic effect of diazepam and xylazine which produced sufficient degree of sedation prior to induction with ketamine. Similarly, Mool (2012) noted induction time of 46.25±1.83 and 30.00±3.27 seconds after diazepam-ketamine and xylazine-ketamine anaesthesia respectively in buffalo calves. Combination of midazolam with ketamine resulted in rapid induction of general anaesthesia characterized by muscle relaxation, analgesia and smooth recovery (Howaida, 2013) as midazolam binds to GABA receptors and decreases the nerve activity. Similar findings were supported by Kumar et al., (2014b) in buffalo calves. Suyawanshi et al., (2023) also reported that diazepam and ketamine facilitate the smooth induction of general anaesthesia in buffalo underwent for diaphragmatic hernia. In the present study, animals of group C showed quicker onset of induction of anaesthesia following ketamine administration which might be due to the synergistic effect of xylazine as it produces sufficient degree of sedation prior to induction with ketamine anaesthesia. These findings are in agreement with Pathak et al., (1982) who reported onset of anaesthesia within 1.75 min after intravenous administration of ketamine anaesthesia only in buffalo calves, probably due to rapid redistribution of ketamine to other body parts.
 
Duration of anaesthesia
 
There was significant (P<0.05) longer duration of anaesthesia in animals of group C as compared to group A and B (Table 1) which might be due to additive effect of xylazine with ketamine completely after induction of anaesthesia with ketamine which signify that surgical stage of anaesthesia has reached in terms of sedation, analgesia and muscle relaxation. All the reflexes abolished. Similarly, Mool (2012) reported longer duration of anaesthesia after xylazine-ketamine (30-34 min) than diazepam-ketamine (20-24 min) in buffalo calves. Avinash et al., (2023) also noted duration of anaesthesia to be 36.00±1.13 after dexmedetomidine-ketamine anaesthesia in bovines. Kumar et al., (2014a) reported duration of anaesthesia in buffalo calves as 37.12 min after diazepam-ketamine anaesthesia respectively. In the present study, diazepam, butorphanol or xylazine was combined with ketamine to prolong the duration of analgesia with good muscle relaxation. Hence, a longer duration of anaesthesia was observed in buffalo calves premedicated with xylazine as compared to butorphanol or diazepam as Pathak et al., (1982) documented that ketamine alone in buffalo calves produced analgesia for 3.0 to 5.50 min (4.45±0.34) with muscular rigidity, thus rendering it practically useless for most surgical procedures.
 
Complete recovery time
 
Complete recovery time was significantly (P<0.05) longer in animals of group C followed by group A and group B. Longer complete recovery time in animals of group C and A might be due to synergistic action of xylazine or diazepam with ketamine whereas shorter recovery in animals of group B revealed faster rate of metabolic clearance of butorphanol-ketamine from the body. Recovery from ketamine anaesthesia occurs through tissue redistribution and hepatic metabolism of the drug making it a short acting anaesthetic drug (Santosh et al., 2013). The present findings corroborates with the findings of Canpolat et al., (2016) and Sengar et al., (2020) who noted recovery time of 108±12.4 min after medetomidine-ketamine anaesthesia and 132.85±3.24 min following medetomidine-ketofol anaesthesia in goats respectively. Similar observations have also been reported by Pawde et al., (2000) and Kumar et al., (2014a) in buffalo calves following detomidine-diazepam-ketamine and diazepam-ketamine anaesthesia respectively.  Correspondingly, Kaur and Singh (2004) also noticed early restoration of vital reflexes and quick recovery after midazolam followed by ketamine in bovines.
 
Physiological parameters
 
Rectal temperature (°F)
 
Animals of group A, B and C exhibited non-significant decrease in rectal temperature after administration of diazepam, butorphanol and xylazine which further tend to decrease significantly (P<0.05) up to 10, 20 and 40 min respectively following ketamine anaesthesia (Fig 1). Later on, these values increased gradually and returned to base value by 120 min of observation. Non significant change in rectal temperature after diazepam or midazolam administration in buffalo calves was reported by Nain et al (2010) which might be attributed to action of benzodiazepine CNS depression and decreased muscular activity alongwith blocking of the hypothalamic thermoregulatory center by the anaesthetics, whereas Almubarak (2013) observed a non significant decrease in rectal temperature after butorphanol administration in camels. The decrease in rectal temperature in group C was also due to activation of α-2 receptor by xylazine and due to depression of thermoregulatory centre in the brain which mediate hypothermia (Lemke, 2004). This might also be resulted due to generalized sedation, reduced metabolic rate, muscle relaxation and CNS depression. Similar findings have also been reported by Yadav et al., (2008) in cattle and Shah et al., (2025) in Barbari goats following xylazine administration. In the present study, decrease in rectal temperature following ketamine anaesthesia could be due to reduced basal metabolic rate, decreased muscle activities leading to production of less heat in the body and depression of hypothalamic thermoregulatory centre of brain (Avinash et al., 2023) and increased in rectal temperature after recovery from ketamine anaesthesia could be attributed to increased tonocity of the muscles. Similar finding were also reported by Canploat et al., (2016) and Rahman et al., (2021) after ketamine anaesthesia in goats and sheep respectively.

Fig 1: Effect on Rectal Temperature (°F) following ketamine anaesthesia in buffalo calves at various time interval in different groups.


 
Respiratory rate (Breaths/min)
 
There was a non-significant increase in respiratory rate in animals of group A, after diazepam administration (Fig 2). However, respiratory rate decreased non significantly by 10 min after induction of ketamine anaesthesia. The present findings support the results reported by Kumar et al., (2014a) after diazepam administration which trend to decrease after induction of ketamine anaesthesia in buffalo calves. Animals of Group B also showed non significant increase in respiratory rate after administration of butorphanol which was further trend to increase non significantly by 10 min after induction with ketamine anaesthesia. Later on the respiratory rate decreased gradually and returned to base value by 120 min of the anaesthetic period. These results concurred with Almubarak (2013) in camels after butorphanol administration. Contrary to this group, Singh et al., (2023) reported a significant (P<0.05) decrease in respiratory rate at 5 min after administration of fentanyl in buffaloes. The respiratory rate was significantly (P<0.05) decreased up to 60 min following xylazine-ketamine administration in group C with maximum reduction at 40 min interval. Later on, the respiratory rate returned to base value by 120 min of the anaesthetic period. The bradypnoea after xylazine-ketamine might be due to depression of CNS by xylazine and due to activation of α2-adrenergic pathway, leading to induction of local coeruleus neurons.  Similar findings were recorded by Venkatgiri et al., (2017) in cattle, Rahman et al., (2021) in sheep and Shah et al., (2025) in Barbari goats. The depression of respiratory rate following administration of xylazine-ketamine might have resulted due to depression of thermoregulatory centre of the brain. Almost all alpha2- agonists were reported to cause some degree of respiration disturbance due to their secondary depression of the CNS (Shah et al., 2014). In the present study, reduced respiration rate might be due to depression of respiratory centers either by xylazine alone or by both xylazine-ketamine.

Fig 2: Effect on Respiratory rate (beaths/min) following ketamine anaesthesia in buffalo calves at various time interval in different groups.


 
Heart rate (Beats/min)
 
There was a non significant decrease in heart rate after administration of diazepam in group A, which tend to increase non-significantly upto 20 min after induction of ketamine anaesthesia (Fig 3). This occurred as a result of diazepam’s parasympatomimetic action and partial baroreceptor reflex desensitization. Similar findings were also pen down by Nain et al., (2010) in buffalo calves. The present findings collaborated with Pawde et al., (2000) and Kumar et al., (2014a) who documented only limited cardiorespiratory effect with no significant changes in heart rate following diazepam-ketamine anaesthesia in buffalo calves. While, animals of group B showed a significant (P<0.05) increase in heart rate after administration of butorphanol which further increased non significantly at 10 min after induction with ketamine and thereafter values gradually returned to base value by 120 min interval. Similar observations were also reported by Almubarak (2013), in camels after butorphanol administration which could be as a result of CNS stimulation. After ketamine induction, increased heart rate might be due to cardiovascular stimulant property of ketamine, which is due to action on sympathetic trunk and inhibition of neuronal uptake of catecholamine by sympathetic nerve endings (Kumar et al., 2014 b) or may be due to increase in central release of catecholamine resulting in tachycardia. There was significant (P<0.05) decrease in heart rate leading to bradycardia after administration of α2-agonist xylazine in group C. The present findings are in concurrent with Pawde et al., (2000) in buffalo calves, Singh et al., (2013) in water buffaloes, Avinash et al., (2023) in bovines and Shah et al., (2025) in Barbari goats. After induction with ketamine anaesthesia heart rate also decreased and then gradually increased and returned to near normal physiological range up to observation period. This might be due to synergistic effect with α2-adrenergic agonist which suppress the cardiovascular stimulating effects of ketamine (Lin and Walz, 2014). The combination of α2- agonists with ketamine has proven suitable for surgical anaesthesia and analgesia. The coadministration of α2-agonist counteracts the muscle rigidity and CNS excitation associated with ketamine while sympathomimetic  effects of ketamine can to some extent offset cardiovascular depressant effects of  α2 agonist (Fazili and Bhattacharyya, 2008). Although ketamine may increase the heart rate due to increased sympathetic activity and decreased vagal tone and xylazine overrides these effects by excitatory carotid baroreceptor reflex induced by hypotension and decreased sympathetic and increased vagal activity (Afshar et al., 2005). On the contrary, Rahman et al., (2021) documented significant (P<0.05) increase in heart rate after atropine-xylazine-ketamine and atropine-ketamine anaesthesia in sheep.

Fig 3: Effect on Heart rate (beats/min) following ketamine anaesthesia in buffalo calves at various time interval in different groups.

CONCLUSION

Present study can be summarized as ketamine in conjunction with xylazine, diazepam and butorphanol proved to be safe general anaesthesia in buffalo calves by providing smooth, safe induction with excitement free recovery that did not result in any adverse effect on cardiopulmonary system. However, combination of the xylazine-ketamine anaesthesia produced longer duration of surgical anaesthesia with smooth recovery in buffalo calves as compared to other two anaesthetic combinations.

ACKNOWLEDGEMENT

The present study was supported by College of Veterinary Science and Animal Husbandry, Anjora (DSVCKV) Durg, Chhattisgarh.
 
Disclaimers
 
The views and conclusions expressed in this article are solely those of the authors and do not necessarily represent the views of their affiliated institutions. The authors are responsible for the accuracy and completeness of the information provided, but do not accept any liability for any direct or indirect losses resulting from the use of this content.
 
Informed consent
 
All animal procedures and handling techniques for experiments were approved by the Institutional Animal Ethical Committee (IAEC). 
 

CONFLICT OF INTEREST

The authors declare that there are no conflicts of interest regarding the publication of this article. No funding or sponsorship influenced the design of the study, data collection, analysis, decision to publish, or preparation of the manuscript.
 

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    Full Research Article

    Evaluation of Clinical and Physiological Parameters Following Ketamine Anaesthesia in Conjunction with Various Premedicants in Male Water Buffalo Calves (Bubalus bubalis)

    K
    Khichar Sangram Singh1
    R
    Rukmani Dewangan1,*
    https://orcid.org/0000-0002-5826-2391
    R
    Raju Sharda1
    J
    Jasmeet Singh1
    I
    Ishant Kumar1
    M
    Muskan Sengar1
    L
    Likchavi Kurrey1
    1Department of Veterinary Surgery and Radiology, College of Veterinary Science and A.H., Dau Shri Vasudev Chandrakar kamdhenu Vishwavidyalaya, Anjora, Durg-491 001, Chhattisgarh, India.

    ABSTRACT

    Background: Ruminants are poor subject for general anaesthesia. With advancement in veterinary sciences, there is need of balanced anaesthesia with rapid, smooth, induction and lesser recumbency time for large animals. Therefore, the present study was planned to evaluate the alternations on clinico-physiological parameters following ketamine anaesthesia in buffalo calves premedicated with either diazepam, butorphanol or xylazine.

    Methods: The study was undertaken in eighteen male buffalo calves and divided into three groups (A, B and C) with six animals in each. Ten minutes prior to the anaesthetic administration, all the buffalo calves were administrated glycopyrrolate 0.01 mg kg b.wt. intramuscularly. The animals of group A, B and C were premedicated intravenously with diazepam 0.5 mg kg b.wt., butorphanol 0.075 mg kg b.wt. and xylazine 0.16 mg kg b.wt. respectively. General anaesthesia was induced with ketamine 4 mg kg b.wt. intravenously and clinical parameters were recorded viz., onset of sedation, induction, duration and recovery of anaesthesia. Physiological parameters were also recorded before (0), 5 min after sedation, induction and at 10, 20, 40, 60 and 120 min following ketamine anaesthesia.

    Result: The onset of sedation and anaesthesia was quicker in animals of group A followed by group C and B respectively. The physiological parameters showed transient changes in all the three groups which were compensated and remained within normal physiological range during the study period. Ketamine in conjunction with various preanaesthetic combinations did not produce any adverse effect on cardiopulmonary system. Thus, ketamine could be safely used as general anaesthestic in buffalo calves premedicated with diazepam, butorphanol or xylazine. However, a combination of xylazine-ketamine produced longer duration of surgical anaesthesia in buffalo calves as compared to other anaesthetic combinations.

    INTRODUCTION

    Ruminants are crucial in India for livelihood and poverty reduction, especially for small and landless farmers who can raise them on marginal lands. Buffalo (Bubalus bubalis) is established as the ‘Black gold’ of India because of most valuable species with excellent zootechnical characteristics for both milk and meat production. However, dairy buffalo rearing can only be rewarding if the buffalo calves are healthy and face less mortality during their early life (Siddhapara et al., 2022). Historically, general anaesthesia was considered unsuitable for ruminants due to various challenges and concerns. Its use was primarily restricted to dogs and horses. Buffaloes are generally considered to be poor subjects to general anaesthesia because of tympany, regurgitation, delayed recovery and haemodynamic disturbances in dorsal and lateral recumbency. There is need for an ideal anaesthetic agent in order to produce desired analgesia, sedation, relaxation and safety for body’s vital system which should also be economical and easy to apply (Pemayun and Sudisma, 2020). Moreover, rapid onset along with quick recovery is desirable in large ruminants due to high risk of regurgitation and tympany. Until now, despite the availability of newer anaesthetic and analgesic drugs, there is no anaesthetic agent that meets the ideal anaesthetic requirement. Therefore, suitable drug combinations are needed to produce a state of balanced anaesthesia. Glycopyrrolate is anti-cholinergic drug which is quaternary ammonium salt and can be administered @ 0.01 mg/kg IM in buffalo calves that blocks cardiac vagus and thereby inhibit cardiac inhibitory effects along with alternation in haemodynamic parameters induced by xylazine in buffalo calves (Potliya et al., 2015). Diazepam is a benzodiazepine derivative and possesses anxiolytic, anticonvulsant, hypnotic, sedative, skeletal muscle relaxant and amnestic properties (Ragab et al., 2022). Butorphanol tartrate is an opioid agonist- antagonist with good analgesic, antitussive and sedative properties which is known to induce only mild sedation and has minimum adverse effects to cardiovascular system in buffalo calves (Bodh et al., 2015). Alpha 2 agonists like xylazine are potent sedatives, widely used in both large and small animals including rapid onset, reliable and potent effect, reversibility and analgesia (Mckelvey and Hollingshead, 2003). Ketamine is a non-competitive N-methyl-d-aspartate receptor antagonist which provides sedation, amnesia with analgesia and has anticonvulsive and neuroprotective properties (Sengar et al., 2020). Ketamine alone produces poor muscle relaxation alongwith muscle spasms. To overcome these undesirable effects, ketamine is often combined with alpha 2  adrenoreceptor agonists like xylazine (Eze et al., 2004), opioids like butorphanol (Bodh et al., 2015; Maidanskaia et al., 2023) and benzodizopines like diazepam (Pawde et al., 2000; Nain et al., 2010) which nullify the potential hypertensive caused by ketamine and providing muscle relaxation. A comprehensive and planned study on the use of ketamine hydrochloride effect in conjunction with various preanaesthetic in buffalo calves are scanty and determining the appropriate general anaesthesia protocol for buffalo due its specific physiological characteristic. Hence, the present study was designed to evaluate the clinico-physiological alternations following ketamine anaesthesia in male buffalo calves premedicated with either diazepam, butorphanol or xylazine.

    MATERIALS AND METHODS

    Anaesthetic protocol
     
    The study was conducted on eighteen healthy male buffalo calves having body weight 80-100 kg and randomly divided into three groups A, B and C comprising six animals in each. The buffalo calves were dewormed with Tab Fenbendazole 150 mg-5 mg/kg body weight orally one month prior to the start of anaesthetic study. Ten minutes prior to the anaesthetic administration, all the buffalo calves were administrated glycopyrrolate 0.01 mg kg b.wt. intramuscularly. The animals of group A, B and C were premedicated intravenously with diazepam 0.5 mg kg b.wt., butorphanol 0.075 mg kg b.wt. and xylazine 0.16 mg kg b.wt. respectively. General anaesthesia was induced with ketamine 4 mg kg b.wt. intravenously and clinical parameters recorded were onset of sedation,  induction, duration and recovery from anaesthesia as per Kumar et al., (2014 a,b) and Bodh et al., (2015). Physiological parameters viz. rectal temperature, heart rate and respiratory rate were recorded at 0 (base value), 5 min after sedation and at 10, 20, 40, 60, 120 min. intervals following ketamine anaesthesia.
     
    Statistical analysis
     
    The data collected were statistically analyzed using Analysis of variance (ANOVA) and Duncan’s Multiple range tests (DMRT) by SPSS v20 statistic software program and all the data were expressed as mean±Standard Error. 

    RESULTS AND DISCUSSION

    Clinical parameters
     
    Onset of sedation
     
    Comparison between groups revealed rapid onset of sedation in animals of Group A than Group C and B. Onset of sedation was significantly (P<0.05) shorter in Group A as compared to Group C and B as shown in Table 1. The shorter period of sedation onset was achieved by administration of diazepam which collaborates with the findings of Kumar et al., (2006) and Nain et al., (2010) in buffalo calves and might be due to depression of CNS that is mediated through depression of limbic system and due to enhancement of gamma-amino butyric acid (GABA) by binding of diazepam to GABAA receptor, an inhibitory channel which when activated, decreases neuronal activity (Hall et al., 2001). In Group C, there was decrease in the behavioural activity of animals after administration of xylazine and thereafter animals went to lateral recumbency within 4.11±1.06 min. Similarly, Nirmale et al., (2024) documented average sedation time 3.75±0.25 min.after administration of xylazine intravenously. Onset of sedation was quicker in this group which could be due to sedative activity of α 2 - adrenoceptor agonists in the locus coeruleus, where noradrenergic neurons are found in high concentration (Singh et al., 2006). These findings are concurrent to Habib et al., (2002) in sheep after intramuscular administration of xylazine and Tunio et al., (2003) in goats after intravenous administration of detomidine. Similarly, Kumar et al., (2006) also reported decrease in spontaneous activity where animals went into sternal recumbency by 8.00±1.00 min following administration of detomidine in buffalo calves. Sedative effects of α-2 agonists are associated with the activation of alpha-2 adrenoceptors, which cause decrease in the release and turnover of norepinephrine in the CNS (Shah et al., 2025). In present study, the administration of butorphanol did not affect degree of sedation and temperament of the animal, as a result longer onset of sedation was noticed in animals of group B. Similar finding was also reported by Almubarak (2013) in camels after administration of butorphanol.

    Table 1: Showing onset of sedation, induction, duration and complete recovery from anaesthesia in different groups.


     
    Induction of anaesthesia
     
    Onset of induction was rapid in animals premedicated with diazepam and xylazine as compared to group B (Table 1), which might be due to synergistic effect of diazepam and xylazine which produced sufficient degree of sedation prior to induction with ketamine. Similarly, Mool (2012) noted induction time of 46.25±1.83 and 30.00±3.27 seconds after diazepam-ketamine and xylazine-ketamine anaesthesia respectively in buffalo calves. Combination of midazolam with ketamine resulted in rapid induction of general anaesthesia characterized by muscle relaxation, analgesia and smooth recovery (Howaida, 2013) as midazolam binds to GABA receptors and decreases the nerve activity. Similar findings were supported by Kumar et al., (2014b) in buffalo calves. Suyawanshi et al., (2023) also reported that diazepam and ketamine facilitate the smooth induction of general anaesthesia in buffalo underwent for diaphragmatic hernia. In the present study, animals of group C showed quicker onset of induction of anaesthesia following ketamine administration which might be due to the synergistic effect of xylazine as it produces sufficient degree of sedation prior to induction with ketamine anaesthesia. These findings are in agreement with Pathak et al., (1982) who reported onset of anaesthesia within 1.75 min after intravenous administration of ketamine anaesthesia only in buffalo calves, probably due to rapid redistribution of ketamine to other body parts.
     
    Duration of anaesthesia
     
    There was significant (P<0.05) longer duration of anaesthesia in animals of group C as compared to group A and B (Table 1) which might be due to additive effect of xylazine with ketamine completely after induction of anaesthesia with ketamine which signify that surgical stage of anaesthesia has reached in terms of sedation, analgesia and muscle relaxation. All the reflexes abolished. Similarly, Mool (2012) reported longer duration of anaesthesia after xylazine-ketamine (30-34 min) than diazepam-ketamine (20-24 min) in buffalo calves. Avinash et al., (2023) also noted duration of anaesthesia to be 36.00±1.13 after dexmedetomidine-ketamine anaesthesia in bovines. Kumar et al., (2014a) reported duration of anaesthesia in buffalo calves as 37.12 min after diazepam-ketamine anaesthesia respectively. In the present study, diazepam, butorphanol or xylazine was combined with ketamine to prolong the duration of analgesia with good muscle relaxation. Hence, a longer duration of anaesthesia was observed in buffalo calves premedicated with xylazine as compared to butorphanol or diazepam as Pathak et al., (1982) documented that ketamine alone in buffalo calves produced analgesia for 3.0 to 5.50 min (4.45±0.34) with muscular rigidity, thus rendering it practically useless for most surgical procedures.
     
    Complete recovery time
     
    Complete recovery time was significantly (P<0.05) longer in animals of group C followed by group A and group B. Longer complete recovery time in animals of group C and A might be due to synergistic action of xylazine or diazepam with ketamine whereas shorter recovery in animals of group B revealed faster rate of metabolic clearance of butorphanol-ketamine from the body. Recovery from ketamine anaesthesia occurs through tissue redistribution and hepatic metabolism of the drug making it a short acting anaesthetic drug (Santosh et al., 2013). The present findings corroborates with the findings of Canpolat et al., (2016) and Sengar et al., (2020) who noted recovery time of 108±12.4 min after medetomidine-ketamine anaesthesia and 132.85±3.24 min following medetomidine-ketofol anaesthesia in goats respectively. Similar observations have also been reported by Pawde et al., (2000) and Kumar et al., (2014a) in buffalo calves following detomidine-diazepam-ketamine and diazepam-ketamine anaesthesia respectively.  Correspondingly, Kaur and Singh (2004) also noticed early restoration of vital reflexes and quick recovery after midazolam followed by ketamine in bovines.
     
    Physiological parameters
     
    Rectal temperature (°F)
     
    Animals of group A, B and C exhibited non-significant decrease in rectal temperature after administration of diazepam, butorphanol and xylazine which further tend to decrease significantly (P<0.05) up to 10, 20 and 40 min respectively following ketamine anaesthesia (Fig 1). Later on, these values increased gradually and returned to base value by 120 min of observation. Non significant change in rectal temperature after diazepam or midazolam administration in buffalo calves was reported by Nain et al (2010) which might be attributed to action of benzodiazepine CNS depression and decreased muscular activity alongwith blocking of the hypothalamic thermoregulatory center by the anaesthetics, whereas Almubarak (2013) observed a non significant decrease in rectal temperature after butorphanol administration in camels. The decrease in rectal temperature in group C was also due to activation of α-2 receptor by xylazine and due to depression of thermoregulatory centre in the brain which mediate hypothermia (Lemke, 2004). This might also be resulted due to generalized sedation, reduced metabolic rate, muscle relaxation and CNS depression. Similar findings have also been reported by Yadav et al., (2008) in cattle and Shah et al., (2025) in Barbari goats following xylazine administration. In the present study, decrease in rectal temperature following ketamine anaesthesia could be due to reduced basal metabolic rate, decreased muscle activities leading to production of less heat in the body and depression of hypothalamic thermoregulatory centre of brain (Avinash et al., 2023) and increased in rectal temperature after recovery from ketamine anaesthesia could be attributed to increased tonocity of the muscles. Similar finding were also reported by Canploat et al., (2016) and Rahman et al., (2021) after ketamine anaesthesia in goats and sheep respectively.

    Fig 1: Effect on Rectal Temperature (°F) following ketamine anaesthesia in buffalo calves at various time interval in different groups.


     
    Respiratory rate (Breaths/min)
     
    There was a non-significant increase in respiratory rate in animals of group A, after diazepam administration (Fig 2). However, respiratory rate decreased non significantly by 10 min after induction of ketamine anaesthesia. The present findings support the results reported by Kumar et al., (2014a) after diazepam administration which trend to decrease after induction of ketamine anaesthesia in buffalo calves. Animals of Group B also showed non significant increase in respiratory rate after administration of butorphanol which was further trend to increase non significantly by 10 min after induction with ketamine anaesthesia. Later on the respiratory rate decreased gradually and returned to base value by 120 min of the anaesthetic period. These results concurred with Almubarak (2013) in camels after butorphanol administration. Contrary to this group, Singh et al., (2023) reported a significant (P<0.05) decrease in respiratory rate at 5 min after administration of fentanyl in buffaloes. The respiratory rate was significantly (P<0.05) decreased up to 60 min following xylazine-ketamine administration in group C with maximum reduction at 40 min interval. Later on, the respiratory rate returned to base value by 120 min of the anaesthetic period. The bradypnoea after xylazine-ketamine might be due to depression of CNS by xylazine and due to activation of α2-adrenergic pathway, leading to induction of local coeruleus neurons.  Similar findings were recorded by Venkatgiri et al., (2017) in cattle, Rahman et al., (2021) in sheep and Shah et al., (2025) in Barbari goats. The depression of respiratory rate following administration of xylazine-ketamine might have resulted due to depression of thermoregulatory centre of the brain. Almost all alpha2- agonists were reported to cause some degree of respiration disturbance due to their secondary depression of the CNS (Shah et al., 2014). In the present study, reduced respiration rate might be due to depression of respiratory centers either by xylazine alone or by both xylazine-ketamine.

    Fig 2: Effect on Respiratory rate (beaths/min) following ketamine anaesthesia in buffalo calves at various time interval in different groups.


     
    Heart rate (Beats/min)
     
    There was a non significant decrease in heart rate after administration of diazepam in group A, which tend to increase non-significantly upto 20 min after induction of ketamine anaesthesia (Fig 3). This occurred as a result of diazepam’s parasympatomimetic action and partial baroreceptor reflex desensitization. Similar findings were also pen down by Nain et al., (2010) in buffalo calves. The present findings collaborated with Pawde et al., (2000) and Kumar et al., (2014a) who documented only limited cardiorespiratory effect with no significant changes in heart rate following diazepam-ketamine anaesthesia in buffalo calves. While, animals of group B showed a significant (P<0.05) increase in heart rate after administration of butorphanol which further increased non significantly at 10 min after induction with ketamine and thereafter values gradually returned to base value by 120 min interval. Similar observations were also reported by Almubarak (2013), in camels after butorphanol administration which could be as a result of CNS stimulation. After ketamine induction, increased heart rate might be due to cardiovascular stimulant property of ketamine, which is due to action on sympathetic trunk and inhibition of neuronal uptake of catecholamine by sympathetic nerve endings (Kumar et al., 2014 b) or may be due to increase in central release of catecholamine resulting in tachycardia. There was significant (P<0.05) decrease in heart rate leading to bradycardia after administration of α2-agonist xylazine in group C. The present findings are in concurrent with Pawde et al., (2000) in buffalo calves, Singh et al., (2013) in water buffaloes, Avinash et al., (2023) in bovines and Shah et al., (2025) in Barbari goats. After induction with ketamine anaesthesia heart rate also decreased and then gradually increased and returned to near normal physiological range up to observation period. This might be due to synergistic effect with α2-adrenergic agonist which suppress the cardiovascular stimulating effects of ketamine (Lin and Walz, 2014). The combination of α2- agonists with ketamine has proven suitable for surgical anaesthesia and analgesia. The coadministration of α2-agonist counteracts the muscle rigidity and CNS excitation associated with ketamine while sympathomimetic  effects of ketamine can to some extent offset cardiovascular depressant effects of  α2 agonist (Fazili and Bhattacharyya, 2008). Although ketamine may increase the heart rate due to increased sympathetic activity and decreased vagal tone and xylazine overrides these effects by excitatory carotid baroreceptor reflex induced by hypotension and decreased sympathetic and increased vagal activity (Afshar et al., 2005). On the contrary, Rahman et al., (2021) documented significant (P<0.05) increase in heart rate after atropine-xylazine-ketamine and atropine-ketamine anaesthesia in sheep.

    Fig 3: Effect on Heart rate (beats/min) following ketamine anaesthesia in buffalo calves at various time interval in different groups.

    CONCLUSION

    Present study can be summarized as ketamine in conjunction with xylazine, diazepam and butorphanol proved to be safe general anaesthesia in buffalo calves by providing smooth, safe induction with excitement free recovery that did not result in any adverse effect on cardiopulmonary system. However, combination of the xylazine-ketamine anaesthesia produced longer duration of surgical anaesthesia with smooth recovery in buffalo calves as compared to other two anaesthetic combinations.

    ACKNOWLEDGEMENT

    The present study was supported by College of Veterinary Science and Animal Husbandry, Anjora (DSVCKV) Durg, Chhattisgarh.
     
    Disclaimers
     
    The views and conclusions expressed in this article are solely those of the authors and do not necessarily represent the views of their affiliated institutions. The authors are responsible for the accuracy and completeness of the information provided, but do not accept any liability for any direct or indirect losses resulting from the use of this content.
     
    Informed consent
     
    All animal procedures and handling techniques for experiments were approved by the Institutional Animal Ethical Committee (IAEC). 
     

    CONFLICT OF INTEREST

    The authors declare that there are no conflicts of interest regarding the publication of this article. No funding or sponsorship influenced the design of the study, data collection, analysis, decision to publish, or preparation of the manuscript.
     

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