Extracorporeal membrane oxygenation improves coagulopathy in an experimental traumatic hemorrhagic model.

PURPOSE: Hemorrhage is the most common cause of preventable death after trauma. Coagulopathy plays a central role in uncontrolled bleeding and is caused by multiple factors. Extracorporeal Membrane Oxygenation (ECMO) is an established treatment for patients with respiratory failure and has in recent years also been used in severely injured trauma patients with cardiopulmonary failure and coexisting bleeding shock. The aim of this study was to evaluate the effect of ECMO on hypothermia, acidosis, and coagulopathy in a traumatic hemorrhagic rabbit model. METHODS: After anesthesia and tracheostomy, ten New Zealand White rabbits sustained laparotomy, bilateral femur fractures and were hemorrhaged 45% of their estimated blood volume. After 90 min of hemorrhagic shock they were resuscitated with a standard transfusion protocol together with venoarterial ECMO (n = 5) or with a standard transfusion protocol only (n = 5) for 60 min. No systemic heparin was administered. RESULTS: ECMO during 60 min of resuscitation significantly increased heart rate (p = 0.01), mean arterial pressure (p = 0.01), body temperature (p = 0.01) and improved the metabolic acidosis, pH (p = 0.01), and lactate (p = 0.01). ECMO also improved the coagulation capacity measured in vitro by Rotational Thromboelastometry with a significant decrease in clot formation time (p < 0.01). This finding was confirmed in vivo with a significant reduction in the animals' ear bleeding time (p < 0.01) and cuticle bleeding time (p < 0.01); 5/5 animals survived in the ECMO group and 3/5 animals survived in the control group. CONCLUSIONS: Heparin-free ECMO stabilizes circulation, improves coagulation, and may impact short-time survival, during the first 60 min, in an experimental traumatic model with severe hemorrhagic shock.

Insulin treatment of streptozotocin-induced diabetes re-establishes the patterns in carbohydrate, fat and amino acid metabolisms in growing pigs.

The effects of streptozotocin (STZ) were studied in eight high-health herd-certified Yorkshire × Swedish Landrace pigs (32.5 ± 2.6 kg initial body weight [BW]), and an insulin treatment protocol was developed to re-establish their metabolisms. A single intravenous dose of 150 mg STZ/kg BW successfully induced hyperglycaemia and alterations in their fat and protein metabolisms. Within 13 h post-STZ treatment blood glucose concentration had fallen to a range of 1.3 to 4.7 mmol/L. Hypoglycaemia was promptly treated with 0.5 g glucose/kg BW intravenously. All the pigs became hyperglycaemic with blood glucose concentrations >23 mmol/L within 48 h post-STZ. Two days post-STZ serum C-peptide concentrations fell below 60 ρmol/L in all the pigs and remained below 96 ρmol/L for five weeks until the end of the study. The pigs were left untreated for one week after STZ injection. At the end of this week 13-fold and nine-fold increases in serum concentrations of triglycerides and non-esterified fatty acids, respectively, were observed. Also, at this time-point a three-fold increase in the concentration of branched-chain amino acids (BCAA) was observed, and alanine and taurine were decreased by approximately 70% and 40%, respectively. During the week when the pigs were untreated, a reduced weight gain was observed, but after the onset of insulin treatment the daily weight gain was at least as good as that of conventional high-health pigs. Then a subcutaneous treatment with short-acting insulin was initiated. The initial dose of 2/3 IU/kg BW daily, divided between two doses, was gradually increased to 1 IU/kg BW. Within three weeks, the insulin treatment restored the metabolic changes in carbohydrate, fat and protein metabolisms produced by the STZ. In conclusion, the results underscore the usefulness of this animal model in translational research as insulin treatment re-establishes the changes in carbohydrate, fat and amino acid metabolisms observed in STZ-diabetic pigs and resolves clinical signs of disease similar to those in humans.

Cartilage repair of experimentally 11 induced osteochondral defects in New Zealand White rabbits.

Articular cartilage has a limited capacity for self-repair in adult humans, and methods used to stimulate regeneration often result in re-growth of fibrous cartilage, which has lower durability. No current treatment option can provide complete repair. The possibility of growth factor delivery into the joint for cartilage regeneration after injury would be an attractive treatment option. A full thickness osteochondral defect of 4 mm in diameter and 2 mm deep was created by mechanical drilling in the medial femoral condyle in 20 female adult New Zealand White rabbits. In an attempt to improve regeneration a hyaluronic hydrogel system, with or without bone morphogenetic protein-2 (BMP-2) was delivered intraarticularly. The contralateral joint defect was treated with saline as control. Throughout the study, rabbits were clinically examined and after 12 (n = 6) or 24 (n = 9) weeks, the rabbits were euthanized and the joints evaluated by histology. The defects healed with fibrocartilage like tissue, and the filling of the defects ranged from less than 25% to complete. The healing of the defects varied both inter- and intra-group wise. Treatment with hyaluronan gel with or without BMP-2 had no effect on cartilage regeneration compared with controls. Instead, severe ectopic bone formation was found in seven joints treated with BMP-2. In conclusion, the present study shows that neither treatment with hyaluronic gel alone, nor in combination with BMP-2, improves the healing of an induced cartilage defect in rabbits. It further shows that BMP-2 can induce ectopic bone formation, which severely affects the functionality of the joint.

Mycotoxins in laboratory rodent feed.

Twenty-one batches of fixed-formula rodent diets from three feed manufacturers were tested for the presence of five mycotoxins: deoxynivalenol (DON), nivalenol (NIV), HT-2 toxin, T-2 toxin and ochratoxin A (OTA). Five batches were also tested for the presence of zearalenone (ZEN) and six batches for aflatoxins. Detectable levels of DON (up to 298 microg/kg), NIV (up to 118 microg/kg), OTA (up to 3.1 microg/kg) or ZEN (up to 26.7 microg/kg) were found in samples from all manufacturers. Three batches contained two (DON or NIV and OTA or ZEN) and one batch contained three (DON, OTA and ZEN) different mycotoxins. Aflatoxins, T-2 and HT-2 were not detected in any of the batches. The concentrations of mycotoxins detected in the feed were low, but indicated that feed ingredients, probably the cereal ingredients, were contaminated by mycotoxins. Since mycotoxins are known to have toxic and/or immunosuppressive effects, non-contaminated ingredients should be used for production of laboratory animal feed. The results imply that an improved quality control of ingredients used for laboratory rodent feed should be implemented.

Anaesthesia with ketamine/medetomidine in the rabbit: influence of route of administration and the effect of combination with butorphanol.

OBJECTIVE: To compare the characteristics of anaesthesia induced with ketamine/medetomidine administered by the subcutaneous and intramuscular routes and to assess the effects of the addition of butorphanol to this combination. STUDY DESIGN: Prospective randomised study. ANIMALS: Six female New Zealand White rabbits. METHODS: Rabbits were given one of four combinations of ketamine and medetomidine (K/M) either subcutaneously (SC) or intramuscularly (IM) on four successive occasions with a 7-day interval between treatments. The dose combinations were; 15/0.25 mg kg-1 SC; 15/0.25 mg kg-1 IM; 15/0.5 mg kg-1 SC, and 15/0.25 mg kg-1 together with 0.4 mg kg-1 butorphanol (K/M/B) SC. The effects of anaesthesia on arterial blood gas values and cardiovascular variables were recorded at predetermined time points. Toe and ear pinch reflexes were judged to determine the duration of surgical anaesthesia. Loss of the righting reflex was used to measure the duration of sleep time. Analyses used repeated measures analysis of variance. RESULTS: All groups lost the righting reflex and ear pinch response. Three animals in the groups that received K/M alone lost their toe pinch reflex, whereas four lost this reflex when given K/M/B. Time of onset of loss of the righting, toe and ear pinch reflexes did not differ significantly among the groups. The higher dose combination of medetomidine with ketamine and the combination of K/M/B produced a greater duration of loss of the ear pinch response than the lower dose of K/M administered by either route. No significant differences were found among the groups in the duration of loss of the toe pinch reflex. All animals developed a moderate bradycardia (mean heart rate <166 beats minute-1) and moderate hypoxaemia (mean PaO2 < 6.0 kPa). Animals given butorphanol showed the greatest reduction in respiratory rate (31 ± 13 breaths minute-1, p < 0.05) but this was not reflected in any significant differences in arterial PCO2, PO2 or pH among the groups. CONCLUSIONS: Administration of K/M by the SC route produced equivalent effects in comparison to intramuscular administration. The addition of butorphanol increased the duration of anaesthesia, but produced a slight increase in the degree of respiratory depression. All dose rates resulted in hypoxaemia so oxygen should be administered when these combinations are used in rabbits. CLINICAL RELEVANCE: Subcutaneous administration is both technically simpler and may cause less discomfort to the animal than IM injection, and so is preferred. The combination of K/M with butorphanol has relatively minor effects on the depth and duration of anaesthesia, so offers little advantage to the use of K/M alone.

Induction of anaesthesia with desflurane and isoflurane in the rabbit.

The characteristics of two techniques of face-mask induction of desflurane anaesthesia (rapid or slow) were compared with the effects of slow isoflurane induction in five New Zealand White (NZW) rabbits. Slow induction used stepwise increments in vapour setting of 2% for desflurane and 0.5% for isoflurane at 30 s intervals. All animals were anaesthetized using each technique according to a randomized block design with one week between treatments. Observations were made of the quality of induction (any struggling or periods of apnoea) and the latency to, and the duration of loss of the righting and toe pinch reflexes recorded. Changes in respiratory rate, arterial blood gas and cardiovascular parameters were also recorded. Induction and recovery times were shorter with rapid desflurane induction in comparison to isoflurane (loss of righting reflex: 139+/-27 s cf. 205+/-48 s), but both techniques were associated with struggling and long periods of apnoea (> 1 min) during the first 4 min after administration. During this period a significant degree of bradycardia, hypercapnia and hypoxaemia occurred with both techniques, but these and the subsequent effects of rapid desflurane administration were less severe than with isoflurane. Slow induction with desflurane was tolerated best, with little or no deleterious behavioural or physiological effects, however excessively prolonged induction times (loss of righting reflex 337+/-160 s) limits the application of this method. Desflurane, administered rapidly, appears to be a more suitable agent than isoflurane. However, as with isoflurane, anaesthesia should only be induced following oxygen supplementation.

Assessment of ketamine/medetomidine anaesthesia in the New Zealand White rabbit.

OBJECTIVE: To compare the characteristics of anaesthesia induced with four dose combinations of ketamine/medetomidine. DESIGN: Prospective randomized study. Animals Five female New Zealand White (NZW) rabbits of approximately 2.3 kg. METHODS: Rabbits were given one of four drug combinations (25/0.25; 15/0.5; 15/0.25 and 10/0.5 mg kg-1 IM) on four successive occasions with a four day interval. Response to injection and then arterial blood gas and cardiovascular parameters were recorded at predetermined time points. Toe and ear pinch reflexes gave measures of total duration of surgical anaesthesia and total sleep time. Analyses used repeated measures analysis of variance. RESULTS: Induction was smooth with little reaction to injection and intubation achieved easily. Two combinations (15/0.25, 10/0.5) produced moderate hypoxaemia (mean pO2 < 8.0 kPa) and two (25/0.25, 15/0.5) very marked hypoxaemia (mean pO2 < 5.3 kPa). This was reversed within 15 minutes of oxygen administration and all rabbits recovered uneventfully. Heart rates fell in all cases, with only minimal effects on arterial blood pressure and no cardiac arrhythmias. Mean duration of surgical anaesthesia was significantly longer for dose groups 25/0.25 (57 ± 12 minutes) and 15/0.5 (59 ± 17 minutes, p = 0.01) compared to dose group 15/0.25 (27 ± 8 minutes). Only three animals in the 10/0.5 mg kg-1 group achieved surgical anaesthesia. Mean duration of loss of the ear pinch reflex was similar between doses, being, respectively, 64 ± 13, 81 ± 7, 60 ± 22 and 62 ± 24 minutes. Sleep time was significantly longer for the 15/0.5 dose (112 ± 10 minutes) compared to 15/0.25 (86 ± 22 minutes, p = 0.04). Sleep times for the 25/0.25 and 10/0.5 mg kg-1 doses were, respectively, 103 ± 23 and 108 ± 12 minutes. CONCLUSIONS: Ketamine/medetomidine reliably produces smooth induction and recovery in the NZW rabbit, but due to the degree of hypoxaemia produced, should only be used with simultaneous provision of oxygen. CLINICAL RELEVANCE: Currently recommended dose rates of ketamine/medetomidine for minor procedures such as ovariohysterectomy in rabbits (25 mg/0.5 mg kg-1) are unnecessarily high; a dose of 15/0.25 mg kg-1 should be adequate for 15-30 minutes of surgical anaesthesia.

Sufentanil and medetomidine anaesthesia in the rat and its reversal with atipamezole and butorphanol.

Injectable anaesthetics are widely used to anaesthetize rats, but recovery times are often prolonged. Reversible anaesthetic regimens have the advantage that animals may be recovered quickly, thus reducing the incidence of postoperative complications such as hypothermia, and also providing a means of treating inadvertent anaesthetic overdose. This study assessed and compared the characteristics of anaesthesia induced with combinations of sufentanil and medetomidine administered as a single subcutaneous or intraperitoneal dose, and reversal with butorphanol and atipamezole. Combinations of sufentanil/medetomidine at 40 microg/150 microg and 50 microg/150 microg/kg administered subcutaneously, and 80 microg/300 microg/kg by intraperitoneal injection were found to produce surgical anaesthesia for 101+/-49, 124+/-45 and 76+/-23 min (means +/- SD) respectively. All three combinations produced marked respiratory depression 30 min after injection (< 50% of resting respiratory rate). Oxygen saturation, measured by pulse oximetry, was < 50% in all groups 30 min following drug administration. Subcutaneous administration is recommended since it resulted in a more reliable and more rapid induction of anaesthesia than intraperitoneal administration. The administration of butorphanol and atipamezole (0.2/0.5 mg/kg s.c.) resulted in a rapid (< 7 min) reversal of anaesthesia and an associated respiratory depression. The induction of anaesthesia with sufentanil/medetomidine and its reversal with a combination of atipamezole and butorphanol is an effective technique for anaesthetizing rats. However, due to the marked respiratory depression and the resulting hypoxia, we recommend that this regimen should only be used in animals which are free from respiratory disease and that oxygen should be provided during anaesthesia.

Effects of repeated anaesthesia with ketamine/medetomidine and of pre-anaesthetic administration of buprenorphine in rats.

Two groups of rats were anaesthetized at weekly intervals for 6 weeks with either ketamine/medetomidine alone (60 mg/0.4 mg/kg i.p.) or ketamine/medetomidine (45 mg/0.3 mg/kg i.p.) one hour following buprenorphine (0.05 mg/kg s.c.). Animals that received buprenorphine had longer periods of surgical anaesthesia (P = 0.04) and a greater depression of both mean pedal withdrawal score (P < 0.01) and mean respiratory rate (P = 0.014). Mean total duration of anaesthesia was also greater in the buprenorphine group on day 1. Sleep times reduced with successive doses of anaesthetic in the buprenorphine group (P = 0.024). Two animals in the buprenorphine group died. Repeated anaesthesia with ketamine/medetomidine alone was not associated with anaesthetic mortality. These results indicate that although buprenorphine has a clear anaesthetic-sparing effect, its use with ketamine/medetomidine may be associated with an increased risk of anaesthetic-related mortality.

Induction of anaesthesia with sevoflurane and isoflurane in the rabbit.

The effects of induction of anaesthesia with sevoflurane and isoflurane were studied in rabbits. All rabbits had periods of apnoea (ranging from 30-180 s) during induction which resulted in moderate hypercapnia and acidosis. Arterial pCO2 rose from 4.1 +/- 0.3 kPa to a peak of 7.6 +/- 0.4 kPa (mean +/- SD) (both agents). All animals showed a significant reduction in heart rate (P < 0.05). Heart rate (HR) fell from 226 +/- 33 to a minimum during induction of 57 +/- 32 (sevoflurane) and 199 +/- 41 to 45 +/- 11 (isoflurane). Most animals struggled violently during induction. Use of sevoflurane did not prevent the breath-holding response seen during induction of anaesthesia with other volatile anaesthetics in this species, and the severe apnoea which occurs may represent a significant hazard. The behaviour of the animals indicated that both sevoflurane and isoflurane are aversive, suggesting that this technique should be avoided whenever possible.