Effects of a Multimodal Approach Using Buprenorphine with/without Meloxicam on Food Intake, Body Weight, Nest Consolidating Behavior, Burrowing Behavior, and Gastrointestinal Tissues in Postoperative Male Mice.

Distress affects animal welfare and scientific data validity. There is a lack of reports on the effects of multimodal analgesic approaches in mice. In this study, under the hypothesis that a multimodal analgesic protocol using buprenorphine with meloxicam has analgesic effects, we evaluated the effects of a multimodal analgesic protocol using buprenorphine with meloxicam on the well-being of mice during analgesic administration by changing the dosage of meloxicam. A total of 42 Slc:ICR male mice were categorized into nonsurgical and surgical groups (7 mice per group) and treated with an anesthetic (isoflurane) and analgesics (buprenorphine ± meloxicam). Analgesics were administered for 48 h after treatment. Buprenorphine (subcutaneous; 0.1 mg/kg/8 h) and meloxicam (subcutaneous; 0, 2.5, or 5 mg/kg/24 h) were administered twice. Body weight, food intake, nest consolidation score, and latency to burrow were evaluated. A significant decrease in food intake was observed 24 h after treatment, while a significant increase was observed 48 h post-treatment in all groups. Body weight showed a decreasing trend but was not significantly reduced. Furthermore, stomach, duodenum, and jejunum tissues showed no morphological abnormalities. Significant differences in burrow diving scores and the latency to burrow were observed between some groups, but these were not regarded as a consequence of the surgery and/or the meloxicam dose. When buprenorphine and meloxicam were combined, administering up to 5 mg/kg/day of meloxicam for 48 h to male mice after abdominal surgery had no significant negative effects on any tested parameters. In conclusion, a multimodal analgesic protocol of buprenorphine with meloxicam is among the options for increasing well-being in mice following abdominal surgery.

Comparison of Analytical Values after Changing to the International Standardized Method for Lactate Dehydrogenase and Alkaline Phosphatase Measurements in Mouse and Rat.

Since April 2020, the method for lactate dehydrogenase (LD) and alkaline phosphatase (ALP) activity measurements in Japan has been switched from the Japan Society of Clinical Chemistry (JSCC) reference method, which is only used in Japan, to the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) reference method. However, in some species, the relationship between the blood values of both enzymes measured by the two methods remains unclear. Hence, values measured by these two methods cannot be used interchangeably. Therefore, this relationship was examined in ICR mice and Wistar/ST rats. The LD and ALP values obtained by both methods were plotted on scatter graphs, and regression equations were obtained. To compare the JSCC (x) and IFCC (y) methods, regression equations were generated for LD values in non-hemolytic samples as follows: y = 0.954x - 4.008 for ICR mice and y = 0.963x - 6.324 for Wistar/ST rats. The conversion factors from the JSCC to the IFCC methods were 0.954 (mice) and 0.963 (rats). The conversion coefficients from the IFCC to the JSCC methods were 1.048 (mice) and 1.088 (rats). For ALP values in fasted mouse and rat samples, the regression equations were y = 0.336x - 2.247 and y = 0.314x - 17.626, respectively. The conversion factors from the JSCC to the IFCC methods were 0.336 (mice) and 0.314 (rats). The conversion coefficients from the IFCC to the JSCC methods were 2.978 (mice) and 3.188 (rats). These conversion factors can be used for the mutual conversion of both measured values during the transition period from the JSCC to the IFCC method. However, it should be noted that the conversion coefficients for both LD and ALP were affected by isozyme composition.

Changes in the blood lactate dehydrogenase measurements in canines and felines following the international standardization of the assay method.

Lactate dehydrogenase (LDH) in blood is measured using the Japanese Society of Clinical Chemistry (JSCC) method in Japan and the International Federation of Clinical Chemistry (IFCC) method in other countries. In human clinical practice, the IFCC method replaced the JSCC method due to international standardization. Moreover, veterinary LDH measurement will also eventually shift to the IFCC method. However, the relationship between the IFCC and JSCC methods for LDH in various animals and whether they can be equated or not have not yet been investigated. This study aimed to present the changes in measurements in canines and felines after switching to the IFCC method. The plasma LDH activity of canines (N=177) and felines (N=115), who visited a secondary care veterinary clinic, was measured using the JSCC and IFCC methods. The IFCC/JSCC ratio was <1.0 in 85% of canines and 88% of felines, indicating that the IFCC method tended to show lower values than the JSCC method, presumably because LDH5 is dominant among the LDH isozymes in canines and felines. The increase in the systematic errors of both assays was in the high value range, with some samples exceeding the error tolerance from near the upper end of the reference range. When switching to the IFCC method for LDH measurement in canines and felines, each institution should consider whether the reference range and clinical diagnostic values established by the JSCC method are appropriate for continued use.

Effects of Multimodal Analgesic Protocol, with Buprenorphine and Meloxicam, on Mice Well-Being: A Dose Finding Study.

The anesthetic or analgesic agent of choice, route and frequency of anesthetic or analgesic administration, and stressors induce distress during the perioperative period. We evaluated a multimodal analgesic protocol using buprenorphine and meloxicam on the well-being of mice. Twenty-four Slc:ICR male mice were divided into control, anesthesia + analgesia, and surgery + anesthesia + analgesia groups. Tap water (orally: PO) and water for injection (subcutaneous: SC) were administered to the control group. Buprenorphine was administered twice (SC, 0.1 mg/kg/8 h) and meloxicam was administered thrice (PO, 5 mg/kg/24 h) to the anesthesia + analgesia and surgery + anesthesia + analgesia groups. The mice were subjected to laparotomy and assessed for several parameters. Even in absence of surgical pain, the anesthesia + analgesia group presented the same negative effects as the surgery + anesthesia + analgesia group. This multimodal analgesic protocol for mice was expected to have an analgesic effect on pain associated with laparotomy but was not sufficient to prevent food intake and weight decrease. This does not negate the need to administer analgesics, but suggests the need to focus on and care not only about the approach to relieve pain associated with surgery, but also other types of distresses to minimize negative side effects that may interfere with postoperative recovery in mice.

Effect of Different Doses of Atipamezole on Reversal of Medetomidine-Induced Tear-Flow Decrease in Rats.

It has been reported that α2-adrenoceptor agonists such as medetomidine decrease tear flow in many species, including rats. Few studies have investigated the involvement of α2-adrenoceptor in decreased tear flow; the issue has not been illustrated sufficiently. Therefore, we aimed to investigate the effect of different doses of atipamezole on the reversal of medetomidine-induced tear-flow decrease to reveal the specific involvement of α2-adrenoceptor. Treatment with 400, 800, or 1600 µg/kg atipamezole (or saline as the control) was intramuscularly administered to rats 15 min following intramuscular administration of 200 µg/kg medetomidine. After medetomidine administration, tear flow was measured using a phenol red thread test (PRTT). PRTT values decreased significantly after 200 µg/kg medetomidine administration. The PRTT values after 800 (optimal dose to reverse) and 1600 µg/kg atipamezole administration reached baseline, but never exceeded it significantly. Treatment with 400 µg/kg atipamezole also reversed the decrease in PRTT value but the PRTT remained lower than baseline. The optimal dose and the higher dose of atipamezole fully reversed the medetomidine-induced decrease in tear flow to the baseline level in rats, while the lower dose of atipamezole partially recovered tear flow.

Effect of Intramuscular Medetomidine Administration on Tear Flow in Rats.

Medetomidine has been reported to decrease tear flow significantly in dogs, cats, and pigs when used as a sedative or analgesic; however, there are no such reports when it comes to rats. The present study aimed to investigate the effect of medetomidine on tear flow in rats. Medetomidine in doses of 50, 100, or 200 µg/kg or a physiological saline solution as the control, were administered intramuscularly to male Slc:Wistar/ST rats. After the administration of medetomidine, tear flow in both eyes was measured using a phenol red thread tear test. The area under the curve (AUC) of phenol red thread test values from baseline to 8 h was calculated. Data were plotted against the dose of medetomidine and simple linear regression analysis was performed. The effect of the drug on phenol red thread test values was considered dose-related when linear analysis yielded a significant relationship. In all medetomidine-treated groups, tear flow decreased significantly in both eyes after administration, while no significant changes were observed in either eye in the control group. The AUC values from baseline to 8 h after administration in groups treated with 100 and 200 µg/kg of medetomidine were significantly lower in both the left and right eyes compared to the control group. The linear regression of the AUC values was significant for both eyes. Our results indicated that the intramuscular administration of medetomidine in rats decreased tear flow significantly in a dose-dependent manner.

Effect of a heat and moisture exchanger on temperature and humidity of inhaled gas in isoflurane-anesthetized dogs.

OBJECTIVE: To investigate the effects of a heat and moisture exchanger (HME) on the temperature and humidity of inhaled gas in isoflurane-anesthetized dogs. STUDY DESIGN: Prospective, interventional study. ANIMALS: A total of four experimental dogs and four client-owned dogs weighing 13.9 ± 7.4 kg (mean ± standard deviation). METHODS: The four experimental dogs were anesthetized on two occasions with and without an intact HME at least 1 week apart. The four client-owned dogs were anesthetized once only for a surgical procedure and assigned to the HME group or no-HME group in alternate order, resulting in six dogs for each group. All dogs were premedicated, anesthetized with propofol and intubated. The HME was connected to the endotracheal tube. Anesthesia was maintained with isoflurane. A digital thermo-hygrometer was placed between the endotracheal tube and HME. The temperature and relative humidity of the inhaled gas were measured every 5 minutes for 60 minutes and the absolute humidity was calculated at each time point. RESULTS: The temperature and absolute humidity of the inhaled gas was significantly higher at 5-60 minutes after intubation in the HME group than in the no-HME group. Absolute humidity was maintained above 29 mg H2O L-1 in the HME group. No significant time-dependent effects on temperature, relative humidity or absolute humidity of the inhaled gas were observed. CONCLUSIONS AND CLINICAL RELEVANCE: The temperature and absolute humidity of the inhaled gas were higher when an HME was used during isoflurane anesthesia in dogs. The use of an HME may reduce the risk of dehydration and dysfunction of the airway mucosal epithelium.

Antiemetic effect of oral maropitant treatment before the administration of brimonidine ophthalmic solution in healthy cats.

OBJECTIVES: The aim of this study was to investigate the antiemetic, behavioural and physiological effects of oral maropitant treatment before the administration of brimonidine ophthalmic solution in healthy cats. METHODS: Five cats received oral maropitant 8 mg or no treatment (control) 18 h before the administration of one drop of brimonidine solution in both eyes. Each cat was administered each of the two treatments, with a washout period of 1 week. The incidence of emesis, retching, sialorrhoea and lip-licking after brimonidine administration was recorded, while behavioural and physiological parameters, including heart rate, mean blood pressure, respiratory frequency and rectal temperature, were recorded before and 0, 30, 60, 90, 120, 180 and 240 mins after brimonidine administration. RESULTS: Emesis and retching were not observed when maropitant was administered. However, 4/5 cats exhibited vomiting and retching in the absence of maropitant pretreatment. The incidence of emesis and retching after brimonidine administration was significantly lower in the treatment group than in the control group. Sialorrhoea occurred in one cat in the control group, while all cats showed lip-licking after brimonidine administration. There were no significant differences in the incidence of sialorrhoea and lip-licking between the two groups. Although behaviour scores were comparable between the two groups, those obtained during heart rate, mean blood pressure and respiratory frequency measurements were significantly lower than the baseline scores; this indicated a sedative effect after brimonidine administration. The heart rate and mean blood pressure significantly decreased after brimonidine administration in both groups, while there were no intergroup differences in the heart rate, mean blood pressure, respiratory frequency and rectal temperature. CONCLUSIONS AND RELEVANCE: Oral maropitant treatment before the administration of brimonidine ophthalmic solution in cats can alleviate emesis and retching without affecting the sedative effects of brimonidine and important physiological parameters.

Effect of medetomidine on tear flow measured by Schirmer tear test I in normal pigs.

Medetomidine, an α2-adrenoceptor agonist, was reported to decrease tear flow in some species. However, there are no reports about the effect of medetomidine on tear flow in pigs. The purpose of this study was to elucidate it. The study was performed in 10 clinically normal female Landrace pigs aged 3 months. Tear flow was measured by the Schirmer tear test (STT) I before (baseline) and 15 and 30 min after intramuscular administration of 80 µg/kg medetomidine. Compared to the STT I value at baseline, the value decreased significantly at 30 min after administration in both the left and right eyes. In pigs treated with medetomidine, an artificial tear solution or ophthalmic gel should be applied to protect the ocular surface.

Effect of intramuscular administration of medetomidine and xylazine on tear flow measured by the Schirmer tear test I in healthy cats.

OBJECTIVES: This study aimed to investigate the effects of intramuscular medetomidine and xylazine on tear flow in healthy cats. METHODS: Five cats each received medetomidine 10, 20, 40 and 80 µg/kg IM; xylazine 1.0, 2.0, 4.0 and 8.0 mg/kg IM; and physiological saline (2.0 ml IM) in a randomised order separated by intervals of at least 1 week. The Schirmer tear test (STT) I was performed in both eyes before and 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7, 8 and 24 h after each dose. RESULTS: The STT I value decreased significantly at 0.5 and 1.0 h and at 0.75 and 1.0 h in both eyes after administration of medetomidine at 10 or 40 µg/kg. After administration of medetomidine 80 µg/kg, there was a significant decrease in the STT I reading at 0.75, 2 and 3 h in the left eye and 0.75, 1, 2 and 3 h in the right eye. The STT I value decreased significantly at: 0.5, 0.75, 1 and 2 h in the left eye and 0.75 h in the right eye after administration of xylazine 1.0 mg/kg; 0.5, 0.75, 1 and 2 h in the left eye and 0.5, 0.75, 1 and 3 h in the right eye after administration of xylazine 2.0 mg/kg; 0.5, 0.75, 1 and 2 h in both eyes after administration of xylazine 4.0 mg/kg; and 0.5, 0.75, 1, 2 and 3 h in the left eye and 0.75, 1, 2, 3 and 4 h in the right eye after administration of xylazine 8.0 mg/kg. CONCLUSIONS AND RELEVANCE: Both medetomidine and xylazine significantly decreased feline tear flow measured by STT I. Therefore, the ocular surface should be monitored carefully and protected appropriately in cats treated with these sedatives.

Sedative and physiological effects of brimonidine tartrate ophthalmic solution in healthy cats.

OBJECTIVE: To determine the effects of brimonidine tartrate ophthalmic solution on sedation, heart rate (HR), respiratory frequency (fR), rectal temperature (RT) and noninvasive mean arterial pressure (MAP) in healthy cats. STUDY DESIGN: Randomized, blinded crossover study, with 1 week washout between treatments. ANIMALS: Six healthy purpose-bred cats. METHODS: Brimonidine tartrate ophthalmic solution 0.1% (one or two drops; 58.6 ± 3.3 µg per drop) or a control solution (artificial tear solution) was administered to six healthy cats. Behavioural observations and measurements of HR, fR, RT and MAP were recorded before and at 30, 60, 90, 120, 180, 240, 300 and 360 minutes after topical administration. Behavioural scores were analysed using Friedman's test for repeated measures to evaluate the time effect in each treatment and treatment effect at each time point. Physiological variables (HR, fR, RT and MAP) were analysed using two-way analysis of variance for repeated measures to evaluate the time and treatment effects. The level of significance was set at p < 0.05. RESULTS: Dose-dependent behavioural and physiological responses were noted. A dose of two drops of brimonidine resulted in sedation in the cats and decreased HR and MAP. Significant sedative effects occurred between 30 and 120 minutes and for physiological responses up to 360 minutes. The most frequent adverse reaction was vomiting, occurring within 40 minutes in all six cats administered two drops and five of the six cats administered one drop of brimonidine. CONCLUSIONS AND CLINICAL RELEVANCE: The results demonstrated that ocular administration of brimonidine 0.1% ophthalmic solution induced sedation in cats and some cardiovascular effects usually associated with α2-adrenoceptor agonists. Further studies should be performed to determine clinical applications for this agent in cats.

Temporal effects of intramuscular administration of medetomidine hydrochloride or xylazine hydrochloride to healthy dogs on tear flow measured by use of a Schirmer tear test I.

OBJECTIVE: To determine the temporal effects on tear flow measurements obtained by use of a Schirmer tear test (STT) I after IM administration of various doses of medetomidine or xylazine to healthy dogs. ANIMALS: 5 healthy purpose-bred male Beagles. PROCEDURES: Each dog received IM injections of 2.0 mL of physiologic saline (0.9% NaCl) solution (control treatment); 0.1% medetomidine hydrochloride (5, 10, 20, and 40 µg/kg), and 2.0% xylazine hydrochloride (0.5, 1.0, 2.0, and 4.0 mg/kg). Treatments were injected into the semimembranosus muscles; there was at least a 1-week interval between successive injections. Order of treatments was determined via a randomized Latin square crossover design. The STT I was performed on both eyes before (baseline) and 0.25, 0.50, 0.75, 1, 2, 3, 4, 5, 6, 7, 8, and 24 hours after each injection. RESULTS: STT I values decreased significantly within 45 minutes after injection of medetomidine or xylazine, which was followed by gradual recovery. The lowest mean STT I value was < 10 mm/min for all sedation treatments, except when dogs received 5 µg of medetomidine/kg. Linear regression of the area under the curve for the 8 hours after administration yielded significant effects for all sedation treatments. CONCLUSIONS AND CLINICAL RELEVANCE: IM administration of medetomidine or xylazine to dogs reduced tear flow in a dose-related manner. Artificial tear solution or ophthalmic ointment should be used to protect the ocular surface when these drugs are administered to dogs.

Slow-down of age-dependent telomere shortening is executed in human skin keratinocytes by hormesis-like-effects of trace hydrogen peroxide or by anti-oxidative effects of pro-vitamin C in common concurrently with reduction of intracellular oxidative stress.

The cellular life-span of cultivated human skin epidermis keratinocytes NHEK-F was shown to be extended up to 150% of population doubling levels (PDLs) by repetitive addition with two autooxidation-resistant derivatives of ascorbic acid (Asc), Asc-2-O-phosphate (Asc2P), and Asc-2-O-alpha-glucoside (Asc2G), respectively, but to be not extended with Asc itself. In contrast, hydrogen peroxide (H(2)O(2)) as dilute as 20 microM which was non-cytotoxic to the keratinocytes, or at 60 microM being marginally cytotoxic achieved the cellular longevity, unexpectedly, up to 160 and 120% of PDLs, respectively, being regarded as a hormesis-like stimulatory effect. The lifespan-extended cells that were administered with Asc2P, Asc2G, or 20 microM H(2)O(2) were prevented from senescence-induced symptoms such as PDL-dependent enlargement of a cell size of 14.7 microm finally up to 17.4 microm upon Hayflick's limit-called loss of proliferation ability as estimated with a channelizer, and retained young cell morphological aspects such as thick and compact shape and intense attachment to the culture substratum even upon advanced PDLs, whereas other non-extended cells looked like thin or fibrous shape and large size upon lower PDLs. The PDL-dependent shortening of telomeric DNA of 11.5 kb finally down to 9.12-8.10 kb upon Hayflick's limit was observed in common for each additive-given cells, but was decelerated in the following order: 20 microM H(2)O(2) > Asc2P = Asc2G > 60 microM H(2)O(2) > Asc = no additive, being in accord with the order of cell longevity. Intracellular reactive oxygen species (ROS) was diminished by Asc2P, Asc2G or 20 microM H(2)O(2), but not significantly by Asc or 60 microM H(2)O(2) as estimated by fluorometry using the redox indicator dye CDCFH. There was no appreciable difference among NHEK keratinocytes that were administered with or without diverse additives in terms of telomerase activity per cell, which was 1.40 x 10(4)-4.48 x 10(4) times lower for the keratinocytes than for HeLa cells which were examined as the typical tumor cells. Thus longevity of the keratinocytes was suggested to be achieved by slowdown of age-dependent shortening of telomeric DNA rather than by telomerase; telomeres may suffer from less DNA lesions due to the continuous and thorough repression of intracellular ROS, which was realized either by pro-vitamin C such as Asc2P or Asc2G that exerted an antioxidant ability more persistent than Asc itself or by 20 microM H(2)O(2) which diminished intracellular ROS assumedly through a hormesis-like effect.

Detection by PCR of the Tyzzer's disease organism (Clostridium piliforme) in feces.

We examined whether the Tyzzer's disease organism, Clostridium piliforme, could be detected in feces by PCR. If the organism could be detected in feces, a diagnosis could be made without sacrifice of the animal. Using the RT strain of C. piliforme, we found that a C. piliforme band could be detected when there were > or = 1 x 10(0) bacteria present in the PCR solution, but the presence of fecal extract in the solution depressed the sensitivity 10 fold. Nevertheless, we could detect the C. piliforme-specific band in fecal extracts from rats in a naturally infected colony, and concluded that the use of PCR to detect C. piliforme DNA in fecal extracts would be a useful diagnostic technique.

Mapping of A gene responsible for cataract formation and its modifier in the UPL rat.

PURPOSE: The Upjohn Pharmaceuticals Limited (UPL) rat is a unique model for cataracts, which are inherited as an autosomal semidominant trait and expressed as early-onset (E-type) cataracts in homozygotes and as late-onset (L-type) cataracts in heterozygotes. In this study, a gene and its modifier, which are responsible for formation of cataract, were mapped. METHODS: Fifty-five BN x (BN x UPL)F(1) backcross rats and 133 BN x UPL intercross rats were produced. The cataracts present in the rats at eye opening were diagnosed as E-type. Cataracts that developed after eye opening were diagnosed as L-type, and the ages when complete opacity in the lens was observed were used as a quantitative trait to map a gene that modifies the development of mature cataracts. Linkage analysis was performed using 64 arbitrarily primed-representational difference analysis (AP-RDA) markers and 74 microsatellite markers. RESULTS: A gene responsible for the formation of cataract was mapped to the vicinity of D2Rat134 on rat chromosome (chr) 2. A candidate gene, connexin 50 (Cx50/Gja8), had a C-to-T transition at codon 340 that is predicted to result in a nonconservative substitution of arginine by tryptophan. Recombination in the Cx50 genotype and formation of cataract was not observed. By quantitative trait loci analysis, a gene that modified the age of the development of mature cataract was mapped on rat chr 5. CONCLUSIONS: A candidate gene for formation of cataracts in UPL rats was mapped to rat chr 2, and the Cx50 gene was a strong candidate. In addition, a potential modifier gene was mapped on chr 5. Future cloning of these genes will provide good targets for new therapies that can delay the progression of cataracts.