Modulation of formalin-induced pain-related behaviour by clonidine and yohimbine in the Speke's hinged tortoise (Kiniskys spekii).

The study was designed to investigate the involvement of noradrenergic and serotonergic receptor systems in the modulation of formalin-induced pain-related behaviour in the Speke's hinged tortoise. Intradermal injection of 100 µL of formalin at a dilution of 12.5% caused pain-related behaviour (hindlimb withdrawal) that lasted for a mean time of 19.28 min (monophasic response). Intrathecal administration of clonidine (α2 -adrenergic receptor agonist) and yohimbine (α2 -adrenergic receptor antagonist) at a dose of 40 µg/kg and 37.5 µg/kg or 50 µg/kg, respectively, caused a highly significant reduction in the duration of the formalin-induced pain-related behaviour. The effect of clonidine was reversed by intrathecal administration of yohimbine at a dose of 26.7 µg/kg. The effect of yohimbine at a dose of 50 µg/kg was reversed by intrathecal injection of 20 µg/kg of the serotonergic receptor antagonist methysergide maleate. When performing antagonistic reactions, the administration of the antagonist was followed immediately by that of the agonist. The study indicates that for experimental purposes, intrathecal route of drug administration through the atlanto-occipital joint is effective in tortoises. The data also suggest that testudines have noradrenergic and serotonergic systems that appear to play a role in the modulation of pain in this species.

Effects of dexamethasone and acetylsalicylic acid on inflammation caused by Complete Freund's adjuvant in the naked mole rat (Heterocephalus glaber).

The naked mole rat (NMR) is a fossorial rodent that has been observed to have a unique nociceptive system in comparison to others. In this study, we explored on characterization of chronic inflammation in the NMR using Complete Freund's adjuvant (CFA) and investigated the effects of dexamethasone and acetylsalicylic acid on the resulting inflammation. The NMRs were injected with 0.1 ml of CFA subcutaneously in the right hind paw, and an equivalent volume of normal saline was injected to the control group. Swelling of the injected right hind limb was observed within 24 h of injection, which involved the tibiotarsal joint, palmar surface and the digits of the injected paw. Swelling persisted for 6 weeks of experimentation and peaked between day 14 and 21. The resulting inflammation affected the mobility, stance and joint rigidity of CFA treated NMRs in comparison to the control group. Treatment of the chronic phase of the inflammation from the 11th day with dexamethasone and acetylsalicylic acid showed no statistical significance in paw circumference compared to the control group, other than on a few, negligible occasions. The present data showed that CFA was able to induce chronic inflammation in the NMR, and the NMR could thus be established as a model for chronic inflammation. There is, however, need for more sensitive parameters to evaluate the effects of anti-inflammatory drugs.

The effects of clonidine and yohimbine in the tail flick and hot plate tests in the naked mole rat (Heterocephalus glaber).

OBJECTIVE: The naked mole rat (NMR) (Heterocephalus glaber) is increasingly considered an important biomedical research model for various conditions like hypoxic brain injury, cancer and nociception. This study was designed to investigate the effects of clonidine and yohimbine, an alpha-2 (α2) adrenoceptor agonist and antagonist respectively in the tail flick and hot plate tests. RESULTS: A significant difference in tail flick latency was noted between saline control and 30 µg/kg clonidine, which was reduced after administration of 30 µg/kg yohimbine. A significant difference in hot plate latency was also noted between saline control and 30 µg/kg clodinine during the periods 30, 45, 60, 75 and 90 min after administration, and between saline control and 10 µg/kg clonidine during 30 min after administration. The hot plate latency by 30 µg/kg clonidine was also reduced by 30 µg/kg yohimbine during 30 min after administration. Since the tail-flick and hot plate tests mediate the effects at spinal and supraspinal levels respectively, the present study indicates the presence and involvement of noradrenergic receptors in thermal antinociception at spinal and supraspinal levels of the NMR, similar to what has been found in other mammals.

Effects of opioids in the formalin test in the Speke's hinged tortoise (Kinixy's spekii).

Little is known about analgesia in lower vertebrates such as the Speke's hinged tortoise (Kinixy's spekii), yet of late they are increasingly being adopted as pets. The effects of morphine (5, 7.5, 10 and 20 mg/kg), pethidine (10, 20, and 50 mg/kg) and naloxone (5 mg/kg) on nociception induced by the formalin test (12.5%, 100 microL) were studied in the Speke's hinged tortoise. Formalin induced a monophasic limb retraction behavioural response and its duration was recorded. The behaviour lasted for 16.4 +/- 0.8 min. Morphine (7.5, 10 and 20 mg/kg) and pethidine (20 and 50 mg/kg) induced significant decrease in the duration of limb retraction in the formalin test. The anti-nociceptive effects were naloxone (5 mg/kg) reversible. The data suggest that the formalin test is a good test for studying nociception and anti-nociception in tortoises and that the opioidergic system plays a role in the control of nociception in these animals.

Investigation of noradrenergic receptor system in anti-nociception using formalin test in the naked mole rat (Heterocephalus glaber).

The naked mole rat (NMR) is a rodent that has gained importance as a biomedical research model for various conditions like hypoxic brain injury, cancer and nociception. This study was designed to investigate possible involvement of the noadrenergic receptor system in antinoception in the NMR, using the alpha-2 adrenergic receptor specific ligands clonidine (agonist) and yohimbine (antagonist) in the formalin test. Formalin test followed 30 min after intraperitoneal administration of ligands or control. A total of 96 naked mole rats were used. A significant reduction in nociceptive behaviours was demonstrated after administration of clonidine in the doses 1,3,10 and 30 µg/kg (n = 8 per group). Doses of clonidine above 30 µg/kg caused loss of motor and proprietion skills exhibited by prostration and failure to turn over when placed on their backs. The antinociception by 3 µg/kg clonidine was reversed by administration of 30 µg/kg of yohimbine. The present study demonstrates that the noradrenergic receptor system is present and involved in formalin test-related antinociceptive mechanisms in the NMR, similar to other mammals. Given the increasing importance of the NMR as a model for pain and nociception, the species may prove useful as an animal model for noradrenergic mechanisms in pain modulation.

Effects of dehydration and heat stress on food intake and dry matter digestibility in East African ruminants.

Comparative investigations were made between wild and domestic ruminants from arid and semi-arid regions and those species from non-arid areas in an attempt to evaluate the adaptations of these ruminants in terms of the effects of heat stress and dehydration on food intake and digestibility. The effect of (a) an intermittent heat load (a daily light cycle of 12 h at 22 degrees C and 12 h at 40 degrees C) compared to 22 degrees C throughout the day and (b) dehydration level of 15% weight loss, with and without the heat load, on the intake and digestibility of a poor quality hay was investigated in the Grant's gazelle, Oryx, the domestic Turkana goats, fat-tailed sheep, zebu cattle, Thomson's gazelle and wildebeest. The intermittent heat load with water available ad libitum depressed the food intake of zebu cattle and Turkana goats by more than 40%. It had no significant effect on the food intake of the other species. The Thomson's and Grants gazelle, oryx, wildebeest and fat-tailed sheep appear well adapted to withstanding a periodic heat load. Dehydration at 22 degrees C caused a marked depression on food intake of all the species investigated. Dehydration together with a heat load caused no further reduction in the food intake by the Grants's gazelle, oryx, and goats but it did cause a further reduction in the intake in the other species. The small non-domestic ruminants (i.e. Grant's and Thomson's gazelle) appear much more digestive efficient than any of their domestic counterpart.

Antinociceptive effects of intrathecal administration of alpha-adrenoceptor antagonists and clonidine in the formalin test in the mouse.

The hypothesis that the descending noradrenergic system tonically inhibits nociception at the spinal level was investigated, using the formalin test in mice. The alpha-adrenoceptor agonist clonidine (0.46 and 0.92 microgram), injected intrathecally, significantly reduced licking activity in both the early and late phase of the test. The alpha 1-antagonist prazosin (3.75, 7.5 and 15 micrograms) and the alpha 2-antagonist yohimbine (7.5 micrograms) also significantly reduced licking activity in both phases. The smaller doses of yohimbine (1.87 and 3.75 micrograms) induced an insignificant reduction of licking in the early phase. Except for the largest doses of clonidine (0.92 microgram), the drugs used had no effect on the general level of activity and motor performance. These results support previous findings that increased noradrenergic activity in the spinal cord inhibits nociception, however, this inhibition seems not to be tonically active. The mechanisms of the antinociceptive actions of alpha-antagonists are not clear.

Effects of codeine, naproxen and dexamethasone on formalin-induced pain in the naked mole-rat.

A dilute solution of formalin (20 microliters of 10% formalin) was injected subcutaneously in the dorsal right hind paw of the naked mole-rat. The injection of the dilute formalin produced two periods of pain behaviour, the early (0-5 minutes) and the late phase (25-60 minutes). These were quantified as the total time spent licking the injected paw. Codeine phosphate (10, 25 or 50 mg kg-1) significantly reduced pain behaviour in both the early and late phase. Codeine administration also induced aggressive, hyperactive behaviour and motor impairment that was naloxone (2 mg kg-1) reversible. Naproxen (200 mg kg-1) and dexamethasone phosphate (30 mg kg-1) significantly reduced licking activity in the late phase only.

Thermal inhibition of nociceptor-driven spinal cord neurones in the cat: a possible neuronal basis for thermal analgesia.

Extracellular and intracellular recordings were made from within the dorsal horn of 10 anaesthetised and gallamine triethiodide-paralysed cats. Inhibition of background and residual noxious-evoked discharge by cooling and warming was demonstrated in 7 out of 33 nociceptor-driven dorsal horn neurones. Five units were inhibited by warming of the noxious mechanical excitatory receptive field. Four units were inhibited by 100%. One unit was inhibited by 42%. Cold stimulation inhibited two units. The background and residual noxious evoked discharge was inhibited by 100%. Cooling (32-20 degrees C) excited two units; warming (32-43 degrees C) also excited two units. Heating above 43 degrees C excited 8 units; cold below 20 degrees C excited 3 units. The units inhibited by thermal stimulation may provide some neuronal basis for thermal analgesia.

The formalin test in the naked mole-rat (Heterocephalus glaber): analgesic effects of morphine, nefopam and paracetamol.

The present experiments were initiated to study the effects of morphine, nefopam and paracetamol in the naked mole-rat, a hairless rodent that lives in subterranean colonies of up to 300, following the inability to demonstrate morphine analgesia in the hot-plate test in the rodent. The formalin test was used. Injection of 20 microliters 10% formalin produced two periods of high licking and pain behaviour, the early (0-5 min) and the late phase (15-60 min). Morphine (10 or 20 mg/kg), nefopam (10 or 20 mg/kg) and paracetamol (200 mg/kg) significantly inhibited the two phases. Paracetamol (400 mg/kg) produced significant analgesia only during the late phase. It is concluded that, unlike in the hot-plate test, it is possible to demonstrate the analgesic effects of morphine in the naked mole-rat, in the formalin test.

Responses of spinal cord neurones to noxious and non-noxious stimulation of the skin and testicle of the rat.

In chloralose-urethane anaesthetized male rats, 24 single units in the lumbar spinal cord which responded to testicular compression were recorded extracellularly with micropipettes. All responded to noxious testicular compression. In addition, 2 units responded to innocuous testicular compression, 10 to noxious cutaneous stimulation and 2 to innocuous warming of the skin. Five units out of 24 had long ascending projections.

Effects of pethidine, acetylsalicylic acid, and indomethacin on pain and behavior in the mole-rat.

The antinociceptive and behavioral effects of pethidine (10, 20, or 30 mg/kg), acetylsalicylic acid (200, 400, or 600 mg/kg) and indomethacin (20, 40, or 50 mg/kg) in the naked mole-rat was studied in the hot-plate test. Instead of inducing analgesia, pethidine caused a dose-dependent reduction in response latency. Sensorimotor impairment and aggressive behavior were also observed following administration of pethidine (20 or 30 mg/kg). All animals receiving pethidine (30 mg/kg) died following fighting when kept in colony cages. Aggressive behavior and death was prevented by naloxone or by keeping animals in single cages. Acetylsalicylic acid (600 mg/kg) and indomethacin (40 or 50 mg/kg) caused a significant increase in response latency. It is concluded that in the mole-rat pethidine elicits aggression, sensorimotor impairment, and apparent hyperalgesia.

Receptive field organisation and electrophysiological responses of spinal cord thermoreactive neurones in the rat.

Receptive fields and electrophysiological responses of seventy-three thermoreactive neurones were studied. The receptive fields were 1 to 10 mm wide and 1 to 15 mm long, for the warm thermoreactive neurones and 5 to 15 mm wide and 2 to 31 mm long for cold thermoreactive neurones. The receptive fields of 5 units excited by warming and heating were 5 to 11 mm wide and 3 to 16 mm long. Six units excited by warming and light mechanical stimulation had receptive fields about 1 to 7 mm wide and 1 to 10 mm long. Those of 3 units excited by cooling and light mechanical stimulation were 3 to 10 mm wide and 3 to 15 mm long. Seven bimodal units had receptive fields that were 2 to 30 mm wide and long. The receptive fields were on the ipsilateral scrotal and or inguinal and perineal skin. Only 1 unit had a bilateral receptive field. Seven dorsal horn neurones showed convergence of warm sensitive and nociceptive afferents. Also, 2 units had convergent inputs from cold sensitive and nociceptive afferents. The noxious mechanical excitatory receptive fields were separate and located on the ipsilateral and contralateral toes, the penis or ipsilateral testicle. The thermal excitatory receptive fields of these units were 15 to 17 mm wide and 20 to 21 mm long. The warm and cold-reactive neurones discharged more with the rise and fall in skin temperature, respectively. Five warm-reactive neurones showed bursting activity. The locations of the thermoreactive neurones studied were similar to those reported earlier. It is concluded that dorsal horn thermoreactive neurones, have mainly ipsilateral receptive fields. Secondly, convergence of temperature sensitive and nociceptive afferents occur in the dorsal horn of the rat.

Morphine induces aggression but not analgesia in the naked mole-rat (Heterocephalus glaber).

1. The antinociceptive effect in the mole-rat of morphine (1, 10, 20 or 30 mg/kg) and nefopam (10 or 20 mg/kg) was studied. 2. In the hotplate test, morphine had no analgesic effect. A reduced response latency after morphine (10 and 20 mg/kg) could possibly be explained by hyperactivity and excited behaviour. 3. After morphine (10, 20 and 30 mg/kg) most of the animals died after fighting when kept in colony cages. Aggressive behaviour and death was prevented by naloxone, or by keeping the animals in single cages. 4. Nefopam (20 mg/kg) significantly increased the latency for the nociceptive response. 5. It was concluded that in the mole-rat, opioid systems in the CNS may not be involved in the regulation of nociception, but in the regulation of agonistic and motor behaviour.

Effects of recombinant human growth hormone on anorexic Nile crocodiles (Crocodylus niloticus).

1. Eleven-month-old Nile crocodiles with poor appetite and retarded growth were injected with 0.325 micrograms/g recombinant human growth hormone (hGH) twice a week for 4 weeks. 2. The treated animals had a mean intake per meal of 29.8 g/kg, while the controls ate only 2.8 g/kg. 3. The treated group gained 8.1% of their initial body weight, while the controls lost 6.3%. 4. During 4 weeks of treatment the body and head length increased by 3.93 and 1.29%, respectively, while no linear growth took place in the controls. 5. The treated group had higher contents of skeletal muscle protein and liver glycogen than the control group. 6. In conclusion, recombinant hGH induces appetite and growth in anorexic crocodiles.

Ruminal escape and apparent degradation of amino acids administered intraruminally in mixtures to cows.

The apparent ruminal degradation and escape of amino acids (AA) administered in 9 different mixtures of essential AA and 8 different mixtures of nonessential AA were studied using two cows fitted with ruminal cannulas. The 600-mmol AA mixtures, which were administered intraruminally using polyethylene glycol as a liquid marker, contained equal amounts of two, four, or eight AA. The amounts of each of the AA in the mixtures were 300, 150, and 75 mmol, respectively. Ruminal degradation and escape were compared with values previously reported for AA administered individually. Across doses, the mean rate of initial degradation (degradation during the 1st h after administration) of essential AA was 26% when the AA were administered in mixtures and 45% when the AA were administered individually. For nonessential AA, the corresponding values were 34 and 54%. Across doses, mean ruminal escape during the first 8 h after essential AA administration was 22% when the AA were administered in mixtures and 16% when the AA were administered individually. For nonessential AA, the corresponding values were 13 and 11%. After intraruminal administration of AA, both individually and in mixtures, significant negative correlations were found between rates of degradation during the 1st h and ruminal escape during an 8-h period. Some AA mixtures caused a net increase in the concentration of other AA in ruminal fluid 1 h after administration. Twelve of the mixtures that did not contain Ala caused a considerable net increase in the concentration of this AA, and 3 AA mixtures containing Arg and Ala caused a marked net increase in the concentration of Trp.