Influence of different anaesthetics on pro-inflammatory cytokine expression in rat spleen

We examined the effect of five anaesthetic drugs commonly used in laboratory animal research (tribromoethanol, ketamine/xylazine, chloral hydrate, pentobarbital, and urethane) on the expression of four pro-inflammatory cytokines. The anaesthetic agents were applied at dosages normally used for deep surgical anaesthesia. Semiquantitative image analysis of interleukin (IL)-1beta, IL-2, IL-6, and tumour necrosis factor alpha (TNFalpha) mRNA expression in the spleen of male Wistar rats 4 h after application of the anaesthetic drugs showed that these had moderate immunomodulatory effects. Ketamine/xylazine, chloral hydrate, and pentobarbital enhanced the basal expression of IL-1beta and IL-6 mRNA in rat spleen, while urethane reduced splenic IL-1beta mRNA expression. Tribromoethanol, ketamine/xylazine, and urethane reduced the basal TNFalpha mRNA levels, whereas TNFalpha mRNA expression was unaffected by chloral hydrate and by pentobarbital. The data demonstrate that these anaesthetics have slight, but significant, effects on the basal immune status of rats.     

Differential effects of chloral hydrate- and ketamine/xylazine-induced anesthesia by the s.c. route

The proper use of anesthetics in animal experimentation has been intensively studied. In this study we compared the use of chloral hydrate (500 mg kg(-1)) and ketamine (167 mg kg(-1)) combined with xylazine (33 mg kg(-1)) by the s.c. route in male Wistar rats. Chloral hydrate and ketamine/xylazine produced a depth of anesthesia and analgesia sufficient for surgical procedures. The decrease of systolic and diastolic blood pressure was of a higher magnitude in rats anesthetized with chloral hydrate than with ketamine/xylazine. The initial microvascular diameter and blood flow velocity did not differ between both agents. On the other hand, ketamine/xylazine reduced the heart rate more intensively than chloral hydrate. Both anesthetics promoted an increase in arterial pCO(2) and a decrease in pH levels compared to unanesthetized animals. The blood glucose levels were of a higher magnitude in rats after ketamine/xylazine anesthesia than after chloral hydrate. In mesenteric arterioles studied in vivo, ketamine/xylazine anesthesia reduced the constrictive effect of noradrenaline and the dilator effect of bradykinin. However, both anesthetics did not modify the vasodilator effect promoted by acetylcholine. Based on our data, we concluded that both anesthetics alter metabolic and hemodynamic parameters, however the use of chloral hydrate in studies of microvascular reactivity in vivo is more appropriate since ketamine/xylazine reduces the responses to vasoactive agents and increases blood glucose levels.     

Four methods for general anaesthesia in the rabbit: a comparative study

The efficacy and safety of pentobarbitone, ketamine/xylazine, fentanyl/fluanisone/diazepam, and halothane/nitrous oxide anaesthesia were compared in 4 groups of six New Zealand White rabbits. Heart and respiratory rates, body temperature, reflexes, blood pressure and blood gases were measured. Pentobarbitone appeared to be unsuitable for anaesthesia in rabbits, as 5 of the 6 rabbits to whom it was administered, required artificial respiration or died. The combinations of ketamine/xylazine and fentanyl-fluanisone/diazepam both produced unpredictable levels of anaesthesia together with a substantial decline in arterial blood pressure and PO2. Despite a severe drop in blood pressure (up to 37.5%), anaesthesia with halothane and nitrous oxide was found to be superior to the other anaesthetic agents.     

Anaesthetic effects of various ratios of ketamine and xylazine in rhesus monkeys (Macacca mulatta).

Intramuscular injection of selected ratios of ketamine and xylazine provided smooth anaesthetic induction, a wide safety margin, and no significant undersirable side effects. Induction and recovery times, duration of anaesthesia, and thermoregulatory ability can be affected by different combinations of ketamine and xylazine. The addition of xylazine to ketamine increases muscle relaxation, recovery time, and duration of anaesthesia, while generally decreasing induction time and thermoregulatory ability.     

Ketamine-xylazine anaesthesia in the Djungarian hamster (Phodopus sungorus)

The combination of ketamine-xylazine was assessed as a surgical anaesthetic in Djungarian hamsters acclimatized to both long (16 h light: 8 h dark) and short (8 h light: 16 h dark) photoperiods. It was concluded that 50 mg/kg of ketamine with 10 mg/kg of xylazine or 100 mg/kg of ketamine with 5-10 mg/kg of xylazine when given together by intraperitoneal injection was a satisfactory general anaesthetic. Two hundred mg/kg of ketamine with 10 mg/kg xylazine caused death in 13 of 24 animals. There were no clinically significant effects on depth of anaesthesia due to photoperiod.     

The effect of ketamine/xylazine and carbon dioxide on plasma luteinizing hormone releasing hormone and testosterone concentrations in the male Norway rat

The effect of a commonly used anaesthetic, ketamine/xylazine and/or carbon dioxide (CO(2)) on plasma luteinizing hormone releasing hormone (LHRH) and testosterone concentrations was determined in male Sprague-Dawley rats. These values were compared with values obtained from pre-anaesthetic control samples. Ketamine/xylazine treatment did not significantly affect testosterone concentrations. In contrast, LHRH started to decrease one hour after ketamine/xylazine administration and continued to significantly decrease after 24 h. In addition, in the CO(2) euthanasia-only group, LHRH concentrations were also significantly decreased. These results suggest that ketamine/xylazine anaesthesia followed by CO(2) euthanasia 24 h later is exerting a significant effect on LHRH concentrations 24 h after anaesthetizing, while only having a slight effect on testosterone, and that CO(2) is exerting an immediate significant effect on LHRH. In conclusion, LHRH analysis should be avoided after ketamine/xylazine anaesthesia and CO(2) euthanasia.     

Effects of anaesthetic agents in interference of naloxone-induced opiate-withdrawal are dose-dependent in opiate-dependent rats

In opiate-dependent rats previous studies showed that anaesthetic agents, such as chloral hydrate, midazolam and ketamine interfere with naloxone-precipitated opiate withdrawal. Each anaesthetic induces a specific pattern of interference, indicating that the interference is agent-dependent. In order to further investigate these effects and highlight a potential pharmacological basis of opiate withdrawal interference through anaesthetic agents, we hypothesized that anaesthetic-mediated interference of opiate withdrawal is also dose-dependent. Three groups of rats were compared in an experimental procedure of rapid withdrawal induction by an antagonist under anaesthesia using sub-anaesthetic dosage of midazolam, ketamine or saline. We observed that sub-anaesthetic dosage of ketamine, or midazolam, interferes significantly with opiate withdrawal expression. This brings arguments in favour of a pharmacological basis underlying rapid antagonists induction in opiate dependent rats.     

An evaluation of three anaesthetic regimes in the gray short-tailed opossum (Monodelphis domestica)

The effect of several anaesthetic agents on the gray short-tailed opossum (Monodelphis domestica) was investigated. Pentobarbitone sodium at a dose of 50 mg/kg sedated the animals but did not produce analgesia or anaesthesia. A combination of ketamine hydrochloride and xylazine at 40 mg/kg and 5 mg/kg, respectively, sedated the animals, but anaesthetic levels were not attained. Halothane was most effective in producing anaesthesia in Monodelphis domestica. Hypothermia was a major side effect with all three anaesthetic regimes.     

Ketamine/xylazine/butorphanol: a new anesthetic combination for rabbits.

Ketamine is often used in combination with tranquilizers to produce surgical anesthesia in rabbits. While generally effective, there is considerable variation in the depth and duration of anesthesia achieved with ketamine combinations. Butorphanol is a mixed agonist-antagonist opioid that is widely used in a variety of other species. In this study, the commonly used ketamine (35 mg/kg)/xylazine (5 mg/kg) combination is compared with ketamine (35 mg/kg)/xylazine (5 mg/kg)/butorphanol (0.1 mg/kg). Rabbits were anesthetized on consecutive weeks with one of the two regimens. Physiologic parameters including heart rate, respiratory rate, blood pressure and arterial blood gases (pH, PO2, PCO2) were measured throughout anesthesia. Loss of palpebral, pedal and righting reflexes were recorded and reflexes were subsequently evaluated. The addition of butorphanol prolonged reflex loss to 140% (X = 68 min +/- 20 SEM) of control for palpebral reflex; 506% (X = 52 min +/- 18 SEM) of control for pedal reflex; and 159% (X = 128 min +/- 21 SEM) of control for righting reflex. Addition of butorphanol to ketamine/xylazine resulted in mild alterations in the physiologic changes traditionally associated with this combination. Butorphanol can be safely added to the ketamine/xylazine combination in rabbits and results in moderate increases in the duration of reflex loss.     

Chronobiological perspectives on myocardial electrophysiological parameters under three types of general anaesthesia in a rat model

The specific aim of the present study, with respect to dependence on the light–dark (LD) cycle under in vivo conditions in spontaneously breathing rats was to review initial state in electrophysiological parameters that may predict the development of heart rhythm disorders in pentobarbital (40 mg/kg), ketamine–xylazine (100 + 15 mg/kg) and zoletil (30 mg/kg) anaesthetized animals. The study was performed using female Wistar rats that were adaptated to an LD cycle (12 h:12 h). Heart rate, PQ and QT intervals were evaluated for their dependence on the LD cycle. The longest PQ interval duration is under zoletil anaesthesia in the light period and the longest QT interval duration is under ketamine–xylazine anaesthesia in both light periods. We concluded that the most significant predisposition toward the development of ventricular arrhythmias originating from disorders of impulse production and conduction occurred under zoletil anaesthesia in the light period; those resulting from disorders in the dispersion of refractory periods occurred under ketamine–xylazine anaesthesia in both the light periods.     

Isoflurane with morphine is a suitable anaesthetic regimen for embryo transfer in the production of transgenic rats

During our initial attempts to produce transgenic rats, we found that an anaesthetic combination typically used for embryo transfer (intramuscular injection of ketamine [90 mg/kg] with xylazine [10 mg/kg]) yielded extensive variation in both the depth and length of anaesthesia. In the present prospective study, we compared the reproductive outcomes afforded by using either isoflurane (5% for induction, 2% for maintenance, carried in 2 l/min of oxygen) with morphine (5 mg/kg s.c., given immediately after isoflurane induction) or ketamine/xylazine in adult (250-300 g), pseudopregnant Sprague-Dawley rats. Each animal was anaesthetized with either isoflurane/morphine or ketamine/xylazine, after which 30 microinjected eggs were transferred into the left uterine horn. The mean pregnancy rate for isoflurane/morphine (15%) was 50% greater than that achieved with ketamine/xylazine (10%). The mean number of live pups (just over five per litter) was comparable for both regimens. All rats given isoflurane/morphine quickly achieved a surgical depth of anaesthesia and experienced a rapid postoperative recovery (3-5 min). In contrast, 25% of rats injected with ketamine/xylazine did not reach a depth of anaesthesia within 10 min that was sufficient for laparotomy, and all that were anaesthetized successfully required an extended postoperative recovery period (60-90 min). These data show that isoflurane/morphine is well tolerated by microinjected embryos and suggest that its use during embryo transfer may provide a means for both reducing the number of pseudopregnant females used and increasing the speed with which rat transgenic projects are completed.     

Tolazoline antagonises ketamine–xylazine anaesthesia in an endangered Black buck (<Emphasis Type="Italic">Antilope cervicapra</Emphasis>)

Seventy-seven anaesthetic events were carried out in 22 captive adult Black bucks (Antilope cervicapra) of either sex with a combination of 2 mg kg⁻¹ ketamine hydrochloride with 0.25 mg kg⁻¹ xylazine hydrochloride using a dart delivered from a blowpipe. Randomised anaesthetised animals received an intravenous injection of either yohimbine hydrochloride (0.125 or 0.25 mg kg⁻¹) or tolazoline hydrochloride (1 or 2 mg kg⁻¹) after 30–40 min of anaesthesia to antagonise the anaesthetic effects. Ketamine–xylazine induced smooth, rapid and reliable anaesthesia within 5–7 min of darting with no clinical adverse effects and causalities during or post-anaesthesia. Yohimbine failed to antagonise the anaesthetic effects of ketamine–xylazine in the Black buck. On the other hand, tolazoline was found to be very effective in hastening recovery in dose-dependent manner within 0.5–1.5 min. This study documents the first report of ketamine–xylazine anaesthesia and its antagonism by tolazoline in captive Black buck.     

The effects of different anaesthetic treatments on the adreno-cortical functions and glucose levels in NZW rabbits

The effects of five anaesthetics on the corticosterone, cortisol and glucose concentrations were investigated in the NZW rabbit. Sixty animals were assigned to 6 treatment groups (n= 10 per group): control ( iv saline solution injection), ketamine (10 mg/kg iv) with either xylazine (3 mg/kg iv) or diazepam (2 mg/kg iv), pentobarbitone (30 mg/kg iv), thiopentone (20 mg/kg iv) and fentanyl/droperidol (1 mg/kg sc). Plasma glucocorticoids were measured by competitive enzymeimmunoassay EIA and glucose by an autoanalyzer, previously validated for this species in both cases. Blood samples were obtained at 6 time-points: before injection, at 10, 30, 60, 120 min and 24 h after injection of the anaesthetics/saline. A significant decrease of plasma glucocorticoids at 10-60 min was observed in the pentobarbitone and fentanyl/ droperidol groups, whereas the administration of ketamine/diazepam or thiopentone stimulated plasma glucocorticoid release, principally in the recovery period. However, in the ketamine/xylazine group no changes were observed in the glucocorticoid levels, except for a significative increase of cortisol at 60-120 min. Glucose levels significantly increased after ketamine/diazepam administration and principally, after ketamine/xylazine treatment. The present data suggest that ketamine/xylazine has little effect on glucocorticoid levels and provides an adequate level of surgical anaesthesia, hence it would be the anaesthetic of choice, although the hyperglycaemic effect after injection has to be considered for any experimental procedures in rabbits.     

Anaesthetics for General Anaesthesia in Growing Pigs

A comparison was made between different anaesthetics for general anaesthesia in growing pigs, with focus on minor surgery under field conditions and for experiments in clinical research. Healthy crossbreed pigs (HampshirexYorkshirexSwedish Landrace) weighing 20–45 kg were used. The anaesthetics combinations compared were 1) azaperone plus metomidate (AM), 2) Zoletil® (zolazepam + tiletamine) plus xylazine (ZX), and 3) Zoletil® plus xylazine plus ketamine (ZXK). Parameters measured were: heart rate, respiratory rate, blood pressure, body temperature, and depth of analgesia (pin-prick). Minor surgery was performed to test the reliability of the “pin-prick” tests. It was clearly shown that AM produces anaesthesia with good cardiovascular stability and is a drug combination that is suitable for minor surgery. ZX also produces a good anaesthesia characterized by reliable and rapid induction. Good cardiovascular function is maintained, and the laryngeal relaxation makes intubation possible. These characteristics are very useful in a laboratory environment, as easy handling to avoid stress is necessary for research. Although it is difficult to evaluate the quality of analgesia from this study, it is concluded that ZX did not provide a superior anasthesia and analgesia compared to AM in crossbreed pigs. However, these drugs are too expensive for regular use in ambulatory practice. The effects of ZXK resemble those of ZX, but the ZXK-drug combination has no anaesthetic advantages and is more laborious to work with. kw|Keywords|k]azaperone; k]metomidate; k]Zoletil®, xylazine; k]ketamine     

Differential Effects of Chloral Hydrate and Pentobarbital Sodium on a Cocaine Level and Its Catecholamine Response in the Medial Prefrontal Cortex: A Comparison with Conscious Rats

Abstract: Adult male Sprague-Dawley rats anesthetized with chloral hydrate and pentobarbital sodium were used as two different treatment groups. Conscious rats were used as a control group. By using baseline (precocaine) concentration as 100%, after cocaine administration (3.0 mg/kg i.v.), the maximal dopamine (DA) increase occurring at the first microdialysis collection period (20 min) in the medial prefrontal cortex was 299 ± 46% for the chloral hydrate group, 168 ± 12% for the pentobarbital sodium group, and 325 ± 23% for the conscious group. At the same time, norepinephrine (NA) increases reached a maximum and were 162 ± 20%, 100 ± 5%, and 141 ± 17%, respectively. The maximal changes of DA and NA in the chloral hydrate group and in the control group were both significantly higher than that in the pentobarbital sodium group. Meanwhile, the cocaine concentration was higher over a 100-min period of time in the chloral hydrate group when compared with the pentobarbital group and the control group. The peak cocaine concentration in dialysate occurred in the same time slot of maximal DA and NA responses, which were 0.65 ± 0.08, 0.30 ± 0.02, and 0.41 ± 0.05 µ M , respectively. Anesthetics suppress the pharmacologic response of neurons, which may explain the difference in catecholamine response between the pentobarbital sodium and the conscious groups. Conversely, because there was no significant difference in DA and NA response between the chloral hydrate group and the conscious group, it may possibly be due to the balancing effect between the higher existing cocaine concentration and the anesthetic suppression on pharmacological response of neurons in the chloral hydrate group. The effect of guide cannula implantation on the cocaine-induced catecholamine response was also evaluated.     

Hypoxia Imaging Using PET and SPECT: The Effects of Anesthetic and Carrier Gas on [64Cu]-ATSM, [99mTc]-HL91 and [18F]-FMISO Tumor Hypoxia Accumulation

Background

Preclinical imaging requires anaesthesia to reduce motion-related artefacts. For direct translational relevance, anaesthesia must not significantly alter experimental outcome. This study reports on the effects of both anaesthetic and carrier gas upon the uptake of [⁶⁴Cu]-CuATSM, [^99mTc]-HL91 and [¹⁸F]-FMISO in a preclinical model of tumor hypoxia.

Methodology/Principal Findings

The effect of carrier gas and anaesthetic was studied in 6 groups of CaNT-bearing CBA mice using [⁶⁴Cu]-CuATSM, [^99mTc]-HL91 or [¹⁸F]-FMISO. Mice were anaesthetised with isoflurane in air, isoflurane in pure oxygen, with ketamine/xylazine or hypnorm/hypnovel whilst breathing air, or in the awake state whilst breathing air or pure oxygen. PET or SPECT imaging was performed after which the mice were killed for organ/tumor tracer quantitation. Tumor hypoxia was confirmed. Arterial blood gas analysis was performed for the different anaesthetic regimes. The results demonstrate marked influences on tumor uptake of both carrier gas and anaesthetic, and show differences between [^99mTc]-HL91, [¹⁸F]-FMISO and [⁶⁴Cu]-CuATSM. [^99mTc]-HL91 tumor uptake was only altered significantly by administration of 100% oxygen. The latter was not the case for [¹⁸F]-FMISO and [⁶⁴Cu]-CuATSM. Tumor-to-muscle ratio (TMR) for both compounds was reduced significantly when either oxygen or anaesthetics (isoflurane in air, ketamine/xylazine or hypnorm/hypnovel) were introduced. For [¹⁸F]-FMISO no further decrease was measured when both isoflurane and oxygen were administered, [⁶⁴Cu]-CuATSM did show an additional significant decrease in TMR. When using the same anaesthetic regimes, the extent of TMR reduction was less pronounced for [⁶⁴Cu]-CuATSM than for [¹⁸F]-FMISO (40–60% versus 70% reduction as compared to awake animals breathing air).

Conclusions/Significance

The use of anaesthesia can have profound effects on the experimental outcome. More importantly, all tested anaesthetics reduced tumor-hypoxia uptake. Anaesthesia cannot be avoided in preclinical studies but great care has to be taken in preclinical models of hypoxia as anaesthesia effects cannot be generalised across applications, nor disease states.     

The effects of anaesthesia and hypothermia on interstitial concentrations of acetylcholine and choline in rat striatum

The effects of the general anaesthetics pentobarbital, chloral hydrate, and halothane on interstitial concentrations of acetylcholine (ACh) in rat striatum were determined using in vivo microdialysis. All 3 anaesthetics decreased ACh. Emergence from anaesthesia coincided with a recovery of ACh to about 80% of basal values. Pentobarbital increased choline in a profile that was the mirror image of ACh. Chloral hydrate had a biphasic effect on choline, consisting of a shortlasting (20 min) initial decrease followed by an increase. When halothane anaesthetized rats were subjected to forced hypothermia by placing them on ice for 30 min, ACh release was further depressed whereas choline was greatly increased. These finding demonstrate that general anaesthetics decrease extracellular concentrations of ACh in the rat striatum and that this effect can be exacerbated by hypothermia.     

A Clinical Evaluation of Four Hypnotic Agents,Using a Latin-Square Design

A double-blind study with a Latin-square design was undertaken on 25 elderly patients, using a placebo and four hypnotic drugs: ethchlorvynol 500 mg., glutethimide 500 mg., chloral hydrate 500 mg., and secobarbital sodium 100 mg. The trial lasted for five weeks. The drugs were all effective compared with the placebo, differences in sleeping time being statistically significant. Differences between these four drugs were not statistically significant. Sleep was induced soonest by secobarbital and ethchlorvynol. Ethchlorvynol and glutethimide had a relatively somewhat longer period of activity than the others. Glutethimide produced most side effects, especially morning drowsiness. Ethchlorvynol and chloral hydrate produced relatively few cases of drowsiness.     

Physiological,pharmacokinetic and liver metabolism comparisons between 3-, 6-, 12- and 18-month-old male Sprague Dawley rats under ketamine-xylazine anesthesia

The main objective of this study was to compare the physiological changes (withdrawal and corneal reflexes, respiratory and cardiac frequency, blood oxygen saturation, and rectal temperature) following intraperitoneal administration of ketamine (80 mg/kg) and xylazine (10 mg/kg) to 3-, 6-, 12- and 18-month-old male Sprague Dawley rats (n=6/age group). Plasma pharmacokinetics, liver metabolism, and blood biochemistry were examined for a limited number of animals to better explain anesthetic drug effects. Selected organs were collected for histopathology. The results for the withdrawal and corneal reflexes suggest a shorter duration and decreased depth of anesthesia with aging. Significant cardiac and respiratory depression, as well as decreased blood oxygen saturation, occurred in all age groups however, cardiac frequency was the most affected parameter with aging, since the 6-, 12-, and 18-month-old animals did not recuperate to normal values during recovery from anesthesia. Pharmacokinetic parameters (T_1/2 and AUC) increased and drug clearance decreased with aging, which strongly suggests that drug exposure is associated with the physiological results. The findings for liver S9 fractions of 18-month-old rats compared with the other age groups suggest that following a normal ketamine anesthetic dose (80 mg/kg), drug metabolism is impaired, leading to a significant increase of drug exposure. In conclusion, age and related factors have a substantial effect on ketamine and xylazine availability, which is reflected by significant changes in pharmacokinetics and liver metabolism of these drugs, and this translates into shorter and less effective anesthesia with increasing age.     

Exposure to warmer postoperative temperatures reduces hypothermia caused by anaesthesia and significantly increases the implantation rate of transferred embryos in the mouse

Embryo transfer (ET) is among the key factors determining the overall efficiency of transgenic technology in the mouse. A successful ET depends among other factors on the quality of the transferred embryos, foster mothers and anaesthetic reagents and on the transfer techniques. Anaesthesia-caused deaths and suboptimal ET procedures are factors which reduce the success of transgenic experiments and mouse colony maintenance. Here we compared the effects of two anaesthetic reagents-a ketamine/xylazine combination, and tribromoethanol (Avertin)-on the rates of implantation and development to term of mouse zygotes transferred into the oviducts of CD-1 foster mothers, and evaluated whether hypothermia caused by anaesthetics after the ET operation could be overcome by postoperative incubation of the foster mothers. We established two experimental groups of fosters, one of which was kept at room temperature (RT, 21 degrees C) with the other in an incubator (33 degrees C) overnight after ET. Rates of implantation, resorption and development to normal fetuses were evaluated by sacrificing the foster mothers on the 15th day of their pregnancy. Our results showed that regardless of the anaesthetic reagents used, the rates of implantation and of development to normal fetuses can be significantly improved by exposing the foster mothers to warmer temperatures (33 degrees C) immediately after the ET operation. These results may have important implications in increasing the success rate of ET with micromanipulated embryos.