Ultrasonographic examination of ocular lesions in farmed halibut.

This paper describes the use of ultrasound to document ocular abnormalities occurring in farmed halibut. A cystic condition of the posterior segment of farmed halibut with a number of ocular sequelae has been reported previously. Further research to elucidate the aetiology and pathogenesis of this condition requires in vivo determination of ocular health over and above that provided by ophthalmoscopy alone. Ocular ultrasonography would appear to be an optimal technique for evaluating the presence of gas- or fluid-filled cysts in the posterior segment and retrobulbar space. The aim of this study was to evaluate this technique for investigation of the piscine eye and specifically to define the role of ultrasonography to detect gas bubble formation in the choroid of farmed halibut, a factor considered central to ocular pathology in many of these fish. In particular the study sought to assess the minimum size of halibut in which intraocular cystic lesions could be identified by ultrasonography and the minimum size of lesion identified by ocular ultrasonography. A second objective was to correlate ultrasonographic findings with lesions found at post mortem ocular examination. A number of halibut of varying sizes, with and without clinically apparent ocular lesions, were subjected to examination under anaesthesia. A proportion of these fish were subsequently sacrificed to allow post mortem ocular evaluation with the aim of correlating ultrasonographic findings with intraocular pathological defects.

Effect of periconceptional undernutrition and gender on hypothalamic-pituitary-adrenal axis function in young adult sheep.

Glucocorticoids are proposed to act as intermediary factors that transcribe the developmental programming sequelae of maternal nutrient restriction (NR). Periconceptional under-nutrition of sheep markedly activates fetal hypothalamic-pituitary-adrenal (HPA) axis activity leading to preterm birth, while transient undernutrition during late gestation in sheep programs adult HPA axis function. To date, no study has examined resting or stimulated HPA axis function in young adult offspring following a periconceptional nutritional challenge. In the present study, 20 ewes were either periconceptionally undernourished (50% metabolisable energy requirements from days 1 to 30 gestation; NR, n = 8) or fed to control levels (100% requirement; controls, n = 12) to term (147 days gestation). Ewes were blood sampled remotely at 2 and 30 days using automated blood sampling equipment. Thereafter, offspring (controls, n = 6/6 males/females; NR, n = 4/4 males/females) were reared to 1 year of age and on separate days received either an i.v. corticotrophin-releasing hormone (CRH; 0.5 microg/kg) and vasopressin (AVP; 0.1 microg/kg) challenge or a synthetic ACTH i.v. bolus (Synacthen; 1.25 microg/kg), and blood samples were taken (manually and remotely) at appropriate intervals for measurement of plasma ACTH and cortisol accordingly. Resting plasma cortisol, assessed remotely, was similar in ewes during undernutrition (control 18.3 +/- 1.4 vs NR 23.4 +/- 1.9 nmol/l) and in offspring at 4 months of age (control male 17.6 +/- 2.9; control female 17.2 +/- 0.4, NR male 16.5 +/- 3.1, NR female 21.7 +/- 4.0 nmol/l). At 12 months of age, however, resting plasma cortisol was significantly increased in NR females (control male 28.0 +/- 1.5, control female 32.9 +/- 9, NR male 32 +/- 7, NR female 53 +/- 10 nmol/l, F 5.7, P = 0.02) despite no difference in plasma ACTH concentration. There was an interaction between nutritional group and gender for both the pituitary and adrenal responses to CRH and AVP, i.e. for controls, females exhibited increased plasma ACTH or cortisol relative to males but for NR this trend was either not present or reversed. The adrenocortical response to synthetic ACTH was gender-dependent only, being greater in female offspring. Combined CRH and AVP provoked a transient hypertension and marked bradycardia in all animals, irrespective of dietary group or gender and could be effectively reproduced by an AVP bolus alone. In conclusion, the present study has shown that periconceptional undernutrition of sheep has only a minor influence on HPA axis function in their young adult offspring when considered alongside the effect of gender per se.

The effect of blood sampling method on indicators of physiological stress in reindeer (Rangifer tarandus tarandus).

The effects of manual blood sampling and remote blood sampling using automatic blood sampling equipment (ABSE) on plasma cortisol and catecholamine concentrations were studied on eight adult female reindeer (Rangifer tarandus tarandus). Contemporary body temperatures and heart rates were also recorded to determine their utility as other possible stress indicators. The animals were blood sampled once every hour with ABSE on 9-10 May and then by manual blood sampling on 13-14 May. Animals were also fitted with equipment to record heart rate and body temperature. Heart rate and body temperature were also recorded continuously without blood sampling on 17-18 May in undisturbed control conditions. Plasma cortisol concentrations were five-to-six fold greater during manual blood sampling compared to sampling with ABSE (F(1,3) = 13.34, P < 0.05). Plasma noradrenaline concentrations were significantly higher (F(1,3) = 22.98, P < 0.05) during manual blood sampling compared to sampling with ABSE, whereas plasma adrenaline concentrations did not differ. Heart rate was higher during manual blood sampling compared to control values. Body temperature was significantly higher during manual sampling compared to values recorded without blood sampling (F(1,4)= 31.65, P < 0.01). In conclusion, plasma cortisol concentration provides an excellent indicator of handling stress in reindeer. The use of ABSE for blood sampling enables measurements of plasma cortisol levels close to basal concentrations that may be used for reference values in studies where indicators of physiological stress are required.

Behavioural responses of red deer to fences of five different designs.

Capercaillie, a large species of grouse, are sometimes killed when they fly into high-tensile deer fences. A fence design which is lower or has a less rigid top section than conventional designs would reduce bird deaths, but such fences would still have to be deer-proof. The short-term behavioural responses of farmed red deer (Cervus elaphus) to fences of five designs, including four that were designed to be less damaging to capercaillie, were measured. Five deer were located on one side of a fence with a larger group (20 animals), from which they had been recently separated, on the other. The efficacy of fences in preventing deer from the small group from rejoining the larger group was also recorded. In addition to a conventional deer fence (C) the four new designs were, an inverted "L" shape (L), a fence with offset electric wire (E), a double fence (D) and a fence with four webbing tapes above (W). Four replicate groups of deer were each tested for 3 days with each fence design. Deer paced the test fence line relatively frequently (a proportion of 0.09 scan observations overall) but significantly less when deer were separated by fences E or C compared to L, W or D (overall difference between fence types, P<0.001). Deer separated by fence E spent significantly more time pacing perimeter fences than deer separated by fences of other types (overall difference between fence types, P<0.01) but deer separated by fence C maintained a low level of fence pacing overall. Analysis of behaviour patterns across the first day and the 3 days of exposure suggested that the novelty of the test fences, rather than the designs per se, influenced the behaviour of the deer. Over the course of the study, no deer crossed either C or L. Three deer crossed E and two deer crossed both W and D. On this basis, field testing, particularly of fence L, would be a useful next step.

GT160-246, a toxin binding polymer for treatment of Clostridium difficile colitis.

GT160-246, a high-molecular-weight soluble anionic polymer, was tested in vitro and in vivo for neutralization of Clostridium difficile toxin A and B activities. Five milligrams of GT160-246 per ml neutralized toxin-mediated inhibition of protein synthesis in Vero cells induced by 5 ng of toxin A per ml or 1.25 ng of toxin B per ml. In ligated rat ileal loops, 1 mg of GT160-246 neutralized fluid accumulation caused by 5 microg of toxin A. At doses as high as 80 mg/loop, cholestyramine provided incomplete neutralization of fluid accumulation caused by 5 microg of toxin A. GT160-246 protected 80% of the hamsters from mortality caused by infection with C. difficile, whereas cholestyramine protected only 10% of animals. Treatment of C. difficile-infected hamsters with metronidazole initially protected 100% of the hamsters from mortality, but upon removal of treatment, 80% of the hamsters had relapses and died. In contrast, removal of GT160-246 treatment did not result in disease relapse in the hamsters. GT160-246 showed no antimicrobial activity in tests with a panel of 16 aerobic bacteria and yeast and 22 anaerobic bacteria and did not interfere with the in vitro activities of most antibiotics. GT160-246 offers a novel, nonantimicrobial treatment of C. difficile disease in humans.

Behavioural and physiological responses of sheep to 16 h transport and a novel environment post-transport.

The effect of a novel lairage environment on the ability of sheep to recover from 16 h of transport was investigated. Sheep were transported from grass paddocks to either novel outside paddocks or inside pens, and housed groups were transported to either familiar or novel inside pens. During transport, sheep from outside paddocks lay down less than those from inside pens. In sheep transported to inside pens, those from outside paddocks spent more time lying and spent less time eating; hay and water intakes during the first 12 h post-transport were lower than those previously kept inside. There was no obvious effect of a novel environment post-transport on blood biochemistry, suggesting that the lower post-transport feed and water intakes in a novel environment did not have a significant effect on the ability of the sheep to recover from the feed and water deprivation associated with transport.

Effects of preslaughter handling on the behaviour, blood biochemistry and carcases of farmed red deer.

Eight groups of six red deer (four groups of males and four groups of females) were held overnight in an unfamiliar paddock, after which they were moved into a holding pen and slaughtered sequentially by stunning with a captive bolt pistol in a restraining pen. On entering the restraining pen, half the deer in each group were stunned immediately, but the other half were stunned after 10 minutes. Deer spent much of the initial period in the holding pen standing stationary in 'alert' postures, but became less alert over time. A significantly higher proportion of males than females were observed in alert 'head-up' postures in the restraining pen. Deer held in the restraining pen for 10 minutes had significantly higher plasma cortisol concentrations than those which were stunned immediately. Females had a significantly higher glycogen concentration postmortem in both the liver and semimembranosus muscle than males, and deer held in the restraining pen for 10 minutes had a significantly lower liver glycogen concentration than deer which were slaughtered immediately. Males had significantly higher semimembranosus muscle pHu values than females, although the differences were small, and of the 14 deer with pHu values above 6-0, 11 were males.

The welfare of deer.

Recent research on the welfare of farmed deer, as for other species, has addressed issues relating to transport, housing and slaughter. While most studies have continued to use a behavioural and physiological approach, new methods of assessing welfare need to consider aspects of cognition and awareness, which are not so easily assessed. It is suggested that a systems-based analysis may be one way to obtain a view of welfare from an animal-centred position. Incorporating an economic perspective could indicate whether consumer choice or legislation would be more likely to lead to higher standards.

Pre-transport loading of farmed red deer: effects of previous overnight housing environment, vehicle illumination and shape of loading race.

The behaviour of farmed red deer was studied while they were being loaded on to a transporter. In experiment 1, the effects of previous overnight housing conditions (indoors, at a space allowance of either 4 or 8 m2 per deer, or in an outdoor raceway) on the ease of loading were investigated. The number of attempts required to load the deer was not significantly affected by their housing conditions or their sex, but there was a significant increase in the number of attempts required after the first day (P < 0.05), suggesting that some aspect of the loading procedure was aversive to the deer. In experiment 2, the effects of illumination inside the vehicle (bright or dim) and the shape of the loading race (straight or curved) were examined. Neither factor significantly influenced the time taken by deer to enter the trailer. However, deer took significantly (P < 0.05) less time to load as the number of trials increased. It is concluded that the loading of deer may be facilitated if the loading raceway is wide enough to allow the deer to move as a group, but narrow enough to prevent the deer from turning round.

HCO3-secretion by bullfrog duodenum: dependence on nutrient Na+ during secretory stimulation.

HCO3(-) secretion across in vitro duodenal mucosa of Rana catesbeiana was investigated under baseline conditions and during secretory stimulation. Baseline secretion was abolished by removal of CO2-HCO3(-)and reduced approximately 60% by removal of nutrient Na+, but was not sensitive to changes in Cl- or K+. Baseline secretion was not directly altered by exposure to 10(-3) M amiloride or 10(-3) M H2DIDS (dihydro-4,4'-diisothiocyanostilbene-2,2'-disulfonic acid) in the nutrient solution and only mildly reduced by acetazolamide. Following removal and restoration of Na+, recovery of secretion was impaired by exposure to acetazolamide (5 x 10(-4) M) or H2DIDS (5 x 10(-4) M) in the nutrient solution. Secretion stimulated by glucagon (10(-6) M) or 16,16-dimethyl prostaglandin E2 (10 microg.mL(-1)) was markedly attenuated by removal of Na+ or by exposure to H2DIDS, but secretion was not altered by acetazolamide (5 x 10(-4) M) or nutrient amiloride (1 mM). Thus, the HCO3(-) that is secreted under nonstimulated conditions derives partly from basolateral Na(+)-dependent uptake and partly from cellular CO2 hydration. Secretagogue-stimulated secretion by duodenal surface epithelium depends on stilbene-sensitive Na+(HCO3(-))n uptake across the basolateral membrane.

Effects of the duration of lairage following transportation on the behaviour and physiology of farmed red deer.

Eight groups of five farmed red deer were transported by road for three hours, after which they were either slaughtered immediately (TO) or held in lairage for three, six or 18 hours (T3, T6 and T18). Liveweight loss increased with lairage time but hot carcase weight was unaffected. Deer spent much of the initial period in lairage standing stationary in 'alert' postures. After eight to 10 hours the proportions of time spent in various postures (standing stationary, moving and lying down) were similar to pre-journey values. None of the blood components associated with dehydration (packed cell volume, osmolality, total protein and sodium) changed significantly with lairage time. Compared with T0 deer, plasma creatine kinase activity was significantly decreased in T18 deer. Lairage time had no effect on skin damage, bruising or muscle glycogen content, although liver glycogen content increased with longer lairage time. Although lairage time had a statistically significant effect on muscle pHu (with T6 deer having the lowest values), the differences were small and none of the carcases had a pHu greater than 6-0.

Lactate dehydrogenase quantification and isoenzyme distribution in physiological response to stress in red deer (Cervus elaphus).

As a contribution to the description of the physiological response to stress in red deer (Cervus elaphus), the potential use of the intracellular glycolytic enzyme, lactate dehydrogenase (LDH) and its isoenzyme LDH-5, as a marker of muscle damage was assessed. The distribution of LDH isoenzymes within red deer tissues was similar to that shown in other ruminants: tissue characterisation showed isoenzyme LDH-5 to be particularly associated with skeletal muscle. High plasma concentrations of creatine kinase, a muscle-specific enzyme, were associated with both high total LDH activity and the percentage of LDH-5 activity in deer undergoing a potentially stressful procedure, transportation followed by simulated abattoir lairage, which is further evidence of the specificity of the isoenzyme for skeletal muscle damage. The activity of LDH was not correlated with the plasma concentration of cortisol, a widely used physiological measure of psychological stress. This may have been due to different time courses for release and the fact that, in the present study, the immediate pre-collection procedures may have induced a short-term stress response.

Effects of bovine somatotrophin (bST) on ovarian function in post-partum beef cows.

The effects of bovine somatotrophin (bST) on ovarian follicle development and function and associated gonadotrophin profiles during the first nine weeks post partum were investigated in beef cows. Thirty-two cows (Shorthorn x Galloway) in moderately low body condition (BC) at calving were fed to maintain BC thereafter. At Weeks 2, 4, 6 and 8 post partum, animals were injected with 320 mg bovine somatotrophin (bST) (T, treated; n = 17) or with the carrier oil only (C, control; n = 15). Ovulation occurred in 4 of 17 T cows and 0 of 15 C cows (P = 0.10) by nine week post partum. Treatment with bST did not affect the numbers of small (3-8 mm in diameter) or large (> 8 mm in diameter) follicles or the granulosa cell populations but enhanced the oestradiol (P < 0.05) and insulin-like growth factor-I (IGF-I) content (P < 0.01) of large follicles by nine weeks post partum. It did not significantly affect the testosterone concentrations of large follicles. Circulating concentrations of growth hormone (GH) and IGF-I were higher in T cows than in C cows (P < 0.001) but were unrelated to gonadotrophin profiles or gonadotrophin receptor concentrations in the follicles. At Week 8, plasma insulin concentrations were higher in T cows than in C cows both before (P < 0.05) and after (P < 0.05) glucose injection. It is concluded that GH may play an important role in mediating the effects of nutritional state on ovarian function during the post-partum period, possibly through alteration of intrafollicular IGF-I concentrations.

In vitro recovery of canine gastric mucosal surface hydrophobicity and potential difference after aspirin damage.

This study determined how the luminal surface hydrophobicity and transmucosal potential difference (PD) of canine gastric mucosa changed during the recovery period after the tissue was challenged with acidified aspirin. Luminal aspirin reduced both the contact angle and PD of mucosae incubated in Ussing chambers. After the removal of aspirin, surface hydrophobicity was found to recover before PD, and nutrient 16,16-dimethyl prostaglandin E2 accelerated the recovery of both parameters. Restoration of luminal surface hydrophobicity may be an important component of how the stomach reestablishes its barrier properties after exposure to a luminal damaging agent.

Johne's disease in a herd of farmed red deer.

An outbreak of Johne's disease in a herd of farmed red deer was studied for four years. Serological, histopathological and cultural techniques were used to monitor the progress of the disease, and delayed type hypersensitivity skin tests were also applied. The results of the serological tests showed that they were poor predictors of future clinical cases and did not consistently identify animals harbouring mycobacteria. The histopathological methods provided a sensitive and specific means of confirming the infection. The skin tests had a low sensitivity and the results were poorly correlated with the serological results in seropositive animals. A vaccination policy was instituted which was accompanied by a change in the pattern of disease. Although the histopathological evidence suggested that the infection was still occurring, there was a marked reduction in the incidence of clinical disease. Vaccinated animals showed a good response to the skin test.

Superovulation in red deer (Cervus elaphus) and Père David's deer (Elaphurus davidianus), and fertilization rates following artificial insemination with Père David's deer semen.

Two comparative studies were undertaken using adult, female red and Père David's deer to examine the ovulatory response of these animals to a superovulation regimen and fertilization rates following inter- and intraspecific laparoscopic insemination. In Expt 1 six Père David's deer and 12 red deer hinds were treated during the breeding season with an intravaginal progesterone-impregnated controlled internal drug release device (CIDR) for 14 days, with 200 iu pregnant mares' serum gonadotrophin (PMSG) administered 72 h before the device was withdrawn and eight injections of ovine FSH given at 12 h intervals starting at the time of PMSG administration. Oestrous behaviour began one day after CIDR device withdrawal (Père David's deer: 24.00 +/- 2.32 h; red deer: 24.60 +/- 2.23 h). The duration of oestrus was greater in Père David's deer than in red deer (17.50 +/- 1.43 h and 8.25 +/- 3.25 h, respectively, P < 0.001). The peak LH surge of Père David's deer was 68.65 +/- 4.74 ng ml-1 occurring 29.00 +/- 2.41 h after removal of the CIDR devices. In comparison, the peak LH surge in red deer was 17.09 +/- 3.64 ng ml-1 (P < 0.001), occurring 24.00 +/- 0.00 h after CIDR device withdrawal. Pre-surge concentrations of LH were also greater (P < 0.001) in Père David's deer (1.37 +/- 0.11 ng ml-1) than in red deer hinds (0.41 +/- 0.02 ng ml-1).(ABSTRACT TRUNCATED AT 250 WORDS)

Cell survival in rabbit gastric glands: effect of extracellular pH, osmolarity, and anoxia.

Although backdiffusion of luminal acid is regarded as a common mechanism of gastric injury, the extracellular pH (pHo) at which cells are irreversibly injured is not well defined. Exclusion of the fluorescent dye propidium iodide was used to estimate cell survival in rabbit gastric glands incubated in buffers of pHo 8.0-2.0. Mean survival (+/- SE) for n = 6 experiments at 2 h in a HEPES buffer of 300 mosM at pHo 8.0, 7.0, 6.0, 4.0, and 2.0 was 80 +/- 3, 91 +/- 2, 90 +/- 2, 71 +/- 2, and 17 +/- 4%, respectively. Survival at acidic pHo was improved in a high KCl buffer: 78 +/- 3 and 38 +/- 7% at pHo 4.0 and 2.0, respectively. Survival in HCO3- buffers was 73 +/- 3, 88 +/- 2, and 92 +/- 3% at pHo 8.0, 7.4, and 6.0. Brief (5 min) exposure to pHo 4.0 followed by reexposure to pHo 7.4 had no effect on acid secretion as estimated by [14C]aminopyrine uptake or cellular viability over 4 h. The influence on cell survival of changes in pHo under conditions of chemical anoxia and in buffers of different osmolarity was investigated. Chemical anoxia was induced using 2.5 mM KCN and 2.0 mM iodoacetic acid (IAA) to inhibit oxidative phosphorylation and anaerobic glycolysis. Survival in glands exposed to KCN and IAA in HEPES buffer was 54 +/- 5, 82 +/- 3, and 87 +/- 2% at pHo 8.0, 7.0, and 6.0.(ABSTRACT TRUNCATED AT 250 WORDS)

Restitution of frog gastric mucosa in vitro: effect of basic fibroblast growth factor.

BACKGROUND: Rapid re-epithelialization after superficial gastric mucosal injury is caused by migration of persisting viable epithelial cells. Basic fibroblast growth factor (bFGF) has been reported to enhance the healing of experimental duodenal ulcer, but its mode of action is unclear. The present experiments examine whether an effect of bFGF on restitution might contribute to such healing. METHODS: Paired halves of bullfrog fundic gastric mucosa in Ussing chambers were injured by luminal exposure to 1 mol/L NaCl for 10 minutes. RESULTS: Luminal protamine or suramin, both known to interfere with endogenous bFGF, significantly inhibited electrophysiological recovery at neutral luminal pH (pHL). Luminal sucrose octasulfate, which prevents acid degradation of bFGF, and an exogenous, acid-resistant form of bFGF allowed electrophysiological recovery at a pHL of 3.0 that completely prevented restitution in control tissues. Electrophysiological recovery correlated well with morphological restitution. The presence of endogenous bFGF in normal and restituting bullfrog mucosa was confirmed by positive staining with a monoclonal antibody. CONCLUSIONS: It is concluded that rapid epithelial repair after surface injury is at least in part mediated by bFGF.