Post-operative nausea and vomiting (PONV) observed in a clinical study designed to assess the analgesic effects of intravenous and subcutaneous methadone in dogs.

Opioids are a key component of multimodal analgesia. Methadone is licensed in Europe for IV, IM and SC use in dogs despite there being no published studies assessing the analgesic efficacy of SC administration. Our intention was to compare the analgesic effect of IV or SC methadone. Fifteen dogs presenting for stifle surgery were administered 0.4 mg/kg methadone IV followed by a randomised 0.0.4 mg/kg methadone IV or SC dose 3 h later. All dogs received ultrasound-guided sciatic and saphenous nerve blocks with bupivacaine prior to surgery. This protocol resulted in opioid adverse effects (hypersalivation, vomiting and/or regurgitation) in 5/15 dogs (33%). Thus, in consultation with the ethical review committee, an otherwise identical protocol using a revised 0.2 mg/kg methadone dose was implemented. In the next three dogs studied, similar opioid adverse effects were found in all three dogs and the study was terminated. This paper highlights the potential for post operative nausea and vomiting (PONV), which may have been induced by methadone when used in combination with efficacious locoregional anaesthesia.

Influence of peri-incisional epaxial muscle infiltration with bupivacaine pre- or post-surgery on opioid administration in dogs undergoing thoraco-lumbar hemilaminectomy.

This study investigated the influence of bupivacaine infiltration before or after hemilaminectomy on peri-operative opioid requirement in dogs. Thirty dogs undergoing T3-L3 hemilaminectomy were randomly assigned to receive peri-incisional infiltration of bupivacaine 2 mg/kg into the epaxial muscles before surgery (Group A), at wound closure (Group B), or no infiltration (Group C). Anaesthesia comprised dexmedetomidine 4 mcg/kg and methadone 0.3 mg/kg IV (premedication), alfaxalone IV (induction), and isoflurane in oxygen (maintenance). All dogs received meloxicam SC/PO prior to induction of general anaesthesia. Response to surgery, defined as a change in autonomic physiological variables >20% above baseline, was treated with fentanyl 2.5 mcg/kg boluses, followed by a continuous rate infusion of fentanyl at 5 mcg/kg/h. The Glasgow Composite Pain Score-Short Form (GCPS-SF) was performed before premedication and at regular intervals until 24 h postoperatively. Methadone 0.2 mg/kg analgesia was given IV if GCPS-SF was ≥5/20. Number of intraoperative, postoperative and total analgesic interventions were recorded. Analgesic interventions were analysed using a chi-squared test using a Pocock approach and statistical significance was set at P < 0.029. The number of intra-operative analgesic interventions in Group A (median, 0; range, 0-2), was significantly lower than in Group B (median, 3; range, 0-5) and Group C (median, 3; range, 0-5; P = 0.019). Regarding postoperative interventions, there were significantly fewer in Group A (median, 0; range, 0-1) and Group B (median, 0; range, 0-1) than in Group C (median, 1; range, 0-2; P = 0.047). Group A (median, 0; range, 0-3), had significantly fewer total analgesic interventions than Group B (median, 3; range, 0-6) and Group C (median, 4; range, 1-7; P = 0.014). Bupivacaine reduced peri-operative opioid administration and pre-surgical peri-incisional infiltration yielded the greatest benefit.

Ice therapy: cool, current and complicated.

This is the fourth Capsule review article provided by the WSAVA Global Pain Council and which discusses the use of ice or cold therapy as a non-pharmacologic modality for pain control in small animal practice. The physiological effects of cold therapy on tissues, receptors and ion channels are discussed; as well as indications, recommendations for, and limitations of use.

Intraperitoneal and incisional analgesia in small animals: simple, cost-effective techniques.

The World Small Animal Veterinary Association Global Pain Council (WSAVA-GPC) has recently published its first "capsule review" by Monteiro et al. These are short articles that present a brief assessment of the scientific evidence and practical recommendations on important, and sometimes controversial, subjects in pain management. The capsules will be published regularly in the Journal of Small Animal Practice, the official journal of the WSAVA. This second article discusses the use of intraperitoneal and incisional analgesia in small animal practice, including their limitations and recommendations by the authors.

Long-term use of non-steroidal anti-inflammatory drugs in cats with chronic kidney disease: from controversy to optimism.

This is the first of a series of capsule reviews published by the World Small Animal Veterinary Association - Global Pain Council (WSAVA-GPC). Each of these short articles provides a brisk assessment of the scientific evidence in specific aspects of pain management, including analgesic techniques, recommendations and controversies surrounding their use. In this first capsule review, the scientific evidence available on the long-term use of non-steroidal anti-inflammatory drugs in cats with concomitant chronic pain and chronic kidney disease is discussed.

Comparative effects of halothane, isoflurane, sevoflurane and desflurane on the electroencephalogram of the rat.

Inhalant anaesthetic agents are commonly used in studies investigating the electroencephalographic (EEG) effects of noxious stimuli in animals. Halothane causes less EEG depression than isoflurane, however, the EEG effects of halothane, isoflurane, sevoflurane and desflurane have not been compared in the same model. This study aimed to compare the EEG effects of these inhalational agents in the rat. Forty male Sprague-Dawley rats were assigned to four groups and anaesthetized with halothane, isoflurane, sevoflurane or desflurane. EEG was recorded from the left and right somatosensory cortices for 5 min at three different multiples of minimal alveolar concentration (MAC) (1.25, 1.5 and 1.75). Median, 95% spectral edge frequency and total power were derived and a single mean value for each was calculated for the first 60 s of each recording period. When the raw EEG contained burst suppression (BS), the BS ratio (BSR) over 60 s was calculated. No BS was found in EEG recorded from the halothane group at any concentration. BS was present at all concentrations with the other anaesthetic agents. BS was almost complete at all concentrations of isoflurane, whereas BSR increased significantly with increasing concentrations of sevoflurane and desflurane. No significant differences were found between the BSR due to the 1.75 MAC multiple of isoflurane, sevoflurane or desflurane. Halothane causes significantly less depression of cortical activity than the newer inhalant agents at equivalent multiples of MAC. These data support the hypothesis that halothane has a fundamentally different mechanism of action than the other inhalant agents.

Application of a modified form of the Glasgow pain scale in a veterinary teaching centre in the Netherlands.

The Glasgow Composite Measure Pain Scale was developed to measure acute pain in dogs in a hospital setting. In this investigation a modified version of the scale was applied in a centre with a different surgical case load and analgesic protocols, and where English is not the first language, to test its validity in a different clinical environment. The modified scale was used to score pain in 60 dogs during the 24 hours after surgery. Their levels of sedation and a clinical impression of their pain were scored at the same time. Three questions were considered; first, how the modified pain score was related to the pain assessed subjectively, secondly, how it related to variables such as the surgical procedure and the dog's health and thirdly, how it changed over time. The mean modified pain scores for the dogs rated subjectively as having no, mild, moderate or severe pain were significantly different, indicating that the modified scale distinguished between pain of different severities. The changes in the dogs' scores also followed the expected changes in their level of pain with time, providing empirical evidence that the scale measures pain.

An investigation of mechanical nociceptive thresholds in dogs with hind limb joint pain compared to healthy control dogs.

This study investigated the effects of osteoarthritis (OA) on somatosensory processing in dogs using mechanical threshold testing. A pressure algometer was used to measure mechanical thresholds in 27 dogs with presumed hind limb osteoarthritis and 28 healthy dogs. Mechanical thresholds were measured at the stifles, radii and sternum, and were correlated with scores from an owner questionnaire and a clinical checklist, a scoring system that quantified clinical signs of osteoarthritis. The effects of age and bodyweight on mechanical thresholds were also investigated. Multiple regression models indicated that, when bodyweight was taken into account, dogs with presumed osteoarthritis had lower mechanical thresholds at the stifles than control dogs, but not at other sites. Non-parametric correlations showed that clinical checklist scores and questionnaire scores were negatively correlated with mechanical thresholds at the stifles. The results suggest that mechanical threshold testing using a pressure algometer can detect primary, and possibly secondary, hyperalgesia in dogs with presumed osteoarthritis. This suggests that the mechanical threshold testing protocol used in this study might facilitate assessment of somatosensory changes associated with disease progression or response to treatment.

Randomised clinical trial comparing clinically relevant sedation outcome measures in dogs after intramuscular administration of medetomidine in combination with midazolam or butorphanol for routine diagnostic imaging procedures.

The aim of this study was to investigate the sedative effects of medetomidine in combination with midazolam or butorphanol for routine imaging procedures in dogs. Eighty client owned dogs were recruited in a prospective, randomised, blinded clinical study and randomly assigned to receive one of four treatments intramuscularly (IM): (1) 30µg/kg medetomidine (Med30); (2) 20µg/kg medetomidine combined with 0.3mg/kg butorphanol (Med20But0.3); (3) 20µg/kg medetomidine combined with 0.3mg/kg midazolam (Med20Mid0.3); and (4) 10µg/kg medetomidine combined with 0.3mg/kg midazolam (Med10Mid0.3). The level of sedation was evaluated using a composite sedation scale assessed by one investigator (0=no sedation, 15=profound sedation). The number of dogs deemed to be adequately clinically sedated and the dose of propofol administered as rescue sedation were recorded. Mean±standard deviation sedation scores at 30min after the commencement of treatment in the groups that received Med20But0.3 (9.8±4) and Med20Mid0.3 (8.9±4.4) were not statistically significantly different from each other, but were significantly different from the group receiving Med10Mid0.3 (5.6±3.6). Only Med20But0.3 was significantly associated with adequate clinical sedation, while Med10Mid0.3 was associated with 85% sedation failure. The rescue sedation dose of propofol (1.5±1mg/kg) for the Med10Mid0.3 group was significantly higher than for other treatments. A sedation score≥10 out of 15 was a satisfactory cut-off to predict adequate clinical sedation. In healthy dogs, the combination of medetomidine with midazolam did not provide comparable sedation to the same dose of medetomidine in combination with butorphanol in a clinical setting.

A comparison between methadone and buprenorphine for perioperative analgesia in dogs undergoing ovariohysterectomy.

OBJECTIVE: To investigate whether preoperative methadone provides superior perioperative analgesia compared to buprenorphine in dogs undergoing ovariohysterectomy. METHODS: Eighty female dogs were recruited to an assessor-blinded, randomised, clinical trial. Dogs received a premedication dose of 0·05 mg/kg acepromazine or 10 µg/kg medetomidine combined with either 0·3 mg/kg methadone or 20 µg/kg buprenorphine intramuscularly. Anaesthesia was induced with propofol and maintained with isoflurane. Pain was assessed using two scoring schemes (a dynamic interactive visual analogue scale and the short form of the Glasgow Composite Pain Scale) before premedication, 30 minutes later and every hour for 8 hours after premedication. If indicated, rescue analgesia was provided with methadone. Meloxicam was administered after the final assessment. The area under the curve for change in pain scores over time and the requirement for rescue analgesia were compared between groups. RESULTS: Groups premedicated with buprenorphine had significantly higher pain scores than those premedicated with methadone. There was no interaction between opioid and sedative for any outcome measure. Rescue analgesia was required by significantly more dogs premedicated with buprenorphine (45%) than that of methadone (20%). CLINICAL SIGNIFICANCE: At the doses investigated, methadone produced superior postoperative analgesia compared to buprenorphine in dogs undergoing ovariohysterectomy.

Comparative Genomics and Mutational Analysis Reveals a Novel XoxF-Utilizing Methylotroph in the Roseobacter Group Isolated From the Marine Environment.

The Roseobacter group comprises a significant group of marine bacteria which are involved in global carbon and sulfur cycles. Some members are methylotrophs, using one-carbon compounds as a carbon and energy source. It has recently been shown that methylotrophs generally require a rare earth element when using the methanol dehydrogenase enzyme XoxF for growth on methanol. Addition of lanthanum to methanol enrichments of coastal seawater facilitated the isolation of a novel methylotroph in the Roseobacter group: Marinibacterium anthonyi strain La 6. Mutation of xoxF5 revealed the essential nature of this gene during growth on methanol and ethanol. Physiological characterization demonstrated the metabolic versatility of this strain. Genome sequencing revealed that strain La 6 has the largest genome of all Roseobacter group members sequenced to date, at 7.18 Mbp. Multilocus sequence analysis (MLSA) showed that whilst it displays the highest core gene sequence similarity with subgroup 1 of the Roseobacter group, it shares very little of its pangenome, suggesting unique genetic adaptations. This research revealed that the addition of lanthanides to isolation procedures was key to cultivating novel XoxF-utilizing methylotrophs from the marine environment, whilst genome sequencing and MLSA provided insights into their potential genetic adaptations and relationship to the wider community.

Use of a transdermal matrix patch of buprenorphine in cats: preliminary pharmacokinetic and pharmacodynamic data.

Six domestic shorthair cats, aged three to four years and weighing 5.1 to 7.4 kg, were used to assess the thermal antinociceptive effect of a transdermal buprenorphine patch, designed to supply 35 mug buprenorphine/hour, which was applied to the shaved thorax. The cats' thermal thresholds were tested before the patch was applied and two, four, six, eight, 10, 12, 14 and 16 hours after it had been applied, and then every six hours until it was removed after 72 hours, and for a further 24 hours afterwards. Blood was collected at each time to measure the plasma concentration of buprenorphine. The patches did not produce a significant change in the thermal thresholds of the cats throughout the testing period. The mean (sd) peak plasma buprenorphine concentration was 10 (0.81) ng/ml.

Genetics and molecular biology of methanotrophs.

Over the last 20 years or so, the obligate methane-oxidizing bacteria (methanotrophs) have attracted considerable interest. As they grow on a relatively cheap and abundant carbon source, they appeared ideal organisms for the production of bulk chemicals, single-cell protein and for use in biotransformations. More recently their cooxidation properties have been investigated for bioremediation, including the removal of chlorinated compounds such as trichloroethylene from polluted groundwaters. These studies have resulted in a great deal of information on the physiology and biochemistry of methanotrophs but sadly the molecular biology and genetic studies of these organisms have lagged behind. This has been in part due to the obligate nature of the methanotrophs and the refractory nature of such organisms to conventional genetic analysis. However, the more recent availability of broad-host range plasmids coupled with improvements in molecular biology methods have allowed the development of molecular genetic techniques for methanotrophs. The purpose of this review is to summarize what is known about the genetics and molecular biology of methanotrophs and how this information can be used to complement previous and current biochemical studies on the unique property of these bacteria, i.e. the ability to oxidize methane to methanol. Recent developments in molecular ecology techniques that may be applied to these apparently ubiquitous organism are also considered.

Refinement of a thermal threshold probe to prevent burns.

Thermal threshold testing is commonly used for pain research. The stimulus may cause burning and merits prevention. Thermal probe modifications hypothesized to reduce burning were evaluated for practicality and effect. Studies were conducted on two humans and eight cats. Unmodified probe 0 was tested on two humans and promising modifications were also evaluated on cats. Probe 1 incorporated rapid cooling after threshold was reached: probe 1a used a Peltier system and probe 1b used water cooling. Probe 2 released skin contact immediately after threshold. Probe 3 (developed in the light of evidence of 'hot spots' in probe 0) incorporated reduced thermal mass and even heating across the skin contact area. Human skin was heated to 48℃ (6℃ above threshold) and the resulting burn was evaluated using area of injury and a simple descriptive scale (SDS). Probe 1a cooled the skin but required further heat dissipation, excessive power, was not 'fail-safe' and was inappropriate for animal mounting. Probe 1b caused less damage than no cooling (27 ± 13 and 38 ± 11 mm(2) respectively, P = 0.0266; median SDS 1.5 and 4 respectively, P = 0.0317) but was cumbersome. Probe 2 was unwieldy and was not evaluated further. Probe 3 produced even heating without blistering in humans. With probe 3 in cats, after opioid treatment, thermal threshold reached cut-out (55℃) on 24 occasions, exceeded 50℃ in a further 32 tests and exceeded 48℃ in the remainder. No skin damage was evident immediately after testing and mild hyperaemia in three cats at 2-3 days resolved rapidly. Probe 3 appeared to be suitable for thermal threshold testing.

A multicentre, prospective, randomised, blinded clinical trial to compare some perioperative effects of buprenorphine or butorphanol premedication before equine elective general anaesthesia and surgery.

REASONS FOR PERFORMING STUDY: Buprenorphine, a µ-agonist opioid, has recently been licensed for equine use, but butorphanol, a κ-agonist opioid, is more commonly used in horses. The effect of the 2 opioids has not previously been compared in a large clinical study. OBJECTIVES: To compare post operative analgesia and physiological variables in horses undergoing elective surgery following premedication with either buprenorphine or butorphanol in a conventional clinical setting. STUDY DESIGN: Multicentre, prospective, randomised, blinded clinical investigation. METHODS: Eighty-nine healthy horses admitted for elective surgery to one of 6 UK equine veterinary clinics were premedicated with acepromazine, a nonsteroidal anti-inflammatory drug, and romifidine followed by intravenous (i.v.) buprenorphine or butorphanol. Anaesthesia was induced with diazepam/ketamine and maintained with isoflurane in oxygen. A range of surgical procedures were performed and supplementary anaesthetic agents given as required. Physiological variables were monitored during anaesthesia and pain, ataxia, sedation and vital function were assessed post operatively. Data were analysed using t-tests, ANOVA, Mann-Whitney U-test and Chi-squared test as appropriate and P<0.05 was regarded as significant, except for multiple comparisons, when P<0.01 was used. RESULTS: Surgery was carried out successfully in all cases and no mortality or serious morbidity occurred. Physiological variables remained within normal limits and all horses recovered successfully, most standing within 1 h of ceasing anaesthesia. There were no significant differences between groups in any variable except post operative pain when scores (simple descriptive scale) between 3 and 6 h were significantly lower after buprenorphine than after butorphanol. CONCLUSIONS: Horses experienced less post operative pain after buprenorphine than after butorphanol premedication. Compared with butorphanol, buprenorphine did not cause any different effects on vital function.

Genome Analysis of Two Pseudonocardia Phylotypes Associated with Acromyrmex Leafcutter Ants Reveals Their Biosynthetic Potential.

The attine ants of South and Central America are ancient farmers, having evolved a symbiosis with a fungal food crop >50 million years ago. The most evolutionarily derived attines are the Atta and Acromyrmex leafcutter ants, which harvest fresh leaves to feed their fungus. Acromyrmex and many other attines vertically transmit a mutualistic strain of Pseudonocardia and use antifungal compounds made by these bacteria to protect their fungal partner against co-evolved fungal pathogens of the genus Escovopsis. Pseudonocardia mutualists associated with the attines Apterostigma dentigerum and Trachymyrmex cornetzi make novel cyclic depsipeptide compounds called gerumycins, while a mutualist strain isolated from derived Acromyrmex octospinosus makes an unusual polyene antifungal called nystatin P1. The novelty of these antimicrobials suggests there is merit in exploring secondary metabolites of Pseudonocardia on a genome-wide scale. Here, we report a genomic analysis of the Pseudonocardia phylotypes Ps1 and Ps2 that are consistently associated with Acromyrmex ants collected in Gamboa, Panama. These were previously distinguished solely on the basis of 16S rRNA gene sequencing but genome sequencing of five Ps1 and five Ps2 strains revealed that the phylotypes are distinct species and each encodes between 11 and 15 secondary metabolite biosynthetic gene clusters (BGCs). There are signature BGCs for Ps1 and Ps2 strains and some that are conserved in both. Ps1 strains all contain BGCs encoding nystatin P1-like antifungals, while the Ps2 strains encode novel nystatin-like molecules. Strains show variations in the arrangement of these BGCs that resemble those seen in gerumycin gene clusters. Genome analyses and invasion assays support our hypothesis that vertically transmitted Ps1 and Ps2 strains have antibacterial activity that could help shape the cuticular microbiome. Thus, our work defines the Pseudonocardia species associated with Acromyrmex ants and supports the hypothesis that Pseudonocardia species could provide a valuable source of new antimicrobials.

Regulation of expression of methane monooxygenases by copper ions.

Many methanotrophs contain both a soluble and a particulate methane monooxygenase. A unique metabolic switch, mediated by copper ions, regulates the expression of these enzymes. When the copper-to-biomass ratio of the cell is low, the soluble enzyme is expressed, and when the copper-to-biomass ratio is high, the particulate enzyme is expressed. A model for the mechanism of this switch is proposed.

Clinical investigation of remifentanil and propofol for the total intravenous anaesthesia of dogs.

Fifteen adult dogs underwent elective ovariectomy. They were premedicated with 0.5 mg/kg methadone and 0.05 mg/kg(-1) atropine administered intramuscularly, and anaesthesia was induced with propofol and maintained with intravenous infusions of remifentanil at 0.6 microg/kg/minute and propofol; the mean (sd) rate of infusion of propofol throughout the period of anaesthesia was 0.33 (0.03) mg/kg/minute. The dogs were ventilated continuously with oxygen while they were anaesthetised. Their haemodynamic parameters were clinically acceptable during the period of anaesthesia. Two dogs received additional atropine to correct bradycardias of less than 60 bpm and several dogs received additional boluses of remifentanil or propofol to maintain an adequate depth of anaesthesia, as determined by a clinical assessment. The mean (range) time to the return of spontaneous respiration after stopping the remifentanil infusion was 11.1 (6.0 to 17.0) minutes, and the mean (range) time to the dogs standing was 38.0 (20.0 to 80.0) minutes. The quality of recovery was good in 12 of the dogs, two showed mild excitation in the immediate postoperative period and the other dog required additional analgesia with methadone.

Spontaneous electroencephalographic changes in a castration model as an indicator of nociception: a comparison between donkeys and ponies.

REASONS FOR PERFORMING STUDY: Donkeys are believed to be less demonstrative of pain than ponies. Research into comparative sensory processing between these species is required to elucidate these behavioural differences. OBJECTIVES: To compare changes in the electroencephalogram (EEG) recorded during castration between donkeys and ponies. STUDY DESIGN: Prospective clinical study. METHODS: Six ponies and 6 donkeys were castrated under halothane anaesthesia after acepromazine premedication and thiopental anaesthetic induction. Markers were inserted into the EEG recording at the time of skin incision (skin) and emasculation (emasc) for both testicles (T1 and T2) during a closed castration. Raw EEG data were analysed and the EEG variables median frequency (F50 ), total power (Ptot ) and spectral edge frequency (F95 ) derived using standard techniques. Baseline values of F50 , Ptot and F95 for each animal were used to calculate the percentage change from baseline at T1skin, T2skin, T1emasc and T2emasc. RESULTS: Decreased F50 values relative to baseline were observed in 4 ponies and 2 donkeys across all castration time points. In the remaining animals, the F50 value increased compared with baseline. Both donkey and pony groups showed an overall decrease in Ptot values compared with baseline at T1skin, but the magnitude of the decrease was significantly less (P = 0.004) in ponies than in donkeys. Donkeys demonstrated an overall greater increase (P = 0.05) in F95 values at T1skin relative to baseline compared with ponies. CONCLUSIONS: Electroencephalographic responses to the noxious stimulus of castration were noted in both donkeys and ponies. Donkeys demonstrated a greater change in Ptot in response to castration than ponies; thus, donkeys appear to demonstrate a cerebral cortical response to a noxious stimulus that is similar to or greater than that in ponies, suggesting that their subtle behavioural expression of pain is not due to a difference in cortical processing of noxious sensory stimuli.