Can novel methods be useful for pain assessment of castrated piglets?

Given that surgical castration is a painful practice performed on millions of pigs every year, a need to identify novel reliable pain assessment tools exists in order to test anaesthetic and analgesic protocols that may reduce related pain. Two treatments were considered: handling (H) and surgical castration (C). Physiological (cortisol, lactate, glycaemia, rectal and eye temperature) and behavioural variables (latency to move after treatment and alterations in posture and walking) were analysed. Cortisol showed the greatest level in C piglets within 20 min after the surgical procedure and a positive correlation with glucose concentration. Eye temperature was higher in C piglets, and the same difference was detected for rectal temperature 3 h after castration. Behavioural parameters revealed that C piglets had longer latency to move and a higher percentage of them showed alterations in posture and walking. Results of this study showed that, in castrated piglets behavioural and physiological alterations occur mainly in the first 3 h from treatment. Latency to move, alterations in posture and walking, and eye temperature appear to give additional and useful information in piglet pain assessment. However, differently from the behavioural parameters considered, eye temperature involves several manipulations of the animals and a long process to acquire the data.     

Survival of Campylobacter jejuni in different gas mixtures

Campylobacter jejuni in fresh chilled chicken meat is known to be a major risk factor for human gastrointestinal disease. In the present study, the survival under chilled conditions of different C. jejuni strains exposed to different gas mixtures usually used for gas packaging of food was examined. Bolton broth and fresh, skinless chicken fillets were inoculated with six and four strains, respectively, and exposed to the gas mixtures 70/30% O(2)/CO(2), 70/30% N(2)/CO(2), and 100% N(2) (the latter only investigated in broth) at refrigeration temperature (4-5 degrees C). In broth culture, the strains survived significantly longer when exposed to 100% N(2) and 70/30% N(2)/CO(2) than in the oxygen-containing gas mixture, 70/30% O(2)/CO(2) (P<0.0001). For the two anaerobic gas mixtures, the reductions only reached 0.3-0.8 log(10) CFU mL(-1) within the same period. In the presence of oxygen, the numbers of C. jejuni were reduced by a minimum of 4.6 log(10) CFU mL(-1) over 21 days. When inoculated onto chicken fillets, the C. jejuni strains also died significantly faster in the oxygen-containing gas mixture, 70/30% O(2)/CO(2) (P<0.0001), reaching reductions of 2.0-2.6 log(10) CFU g(-1) after 8 days. In the gas mixture without oxygen (70/30% N(2)/CO(2)), no reductions were observed.     

Hyperpnea-induced production of TBHP-initiated chemiluminescence in guinea pigs

Antioxidants attenuate hyperpnea-induced airway constriction. It was hypothesized that this type of airway constriction is closely related to reactive oxygen species (ROS). However, there is no direct evidence of an increase in ROS during or right after the course of hyperpnea. To detect ROS production induced by hyperpnea, forty one guinea pigs were divided into four groups: control; control with 95% O2-5% CO2; hyperpnea with 95% air-5% CO2; and hyperpnea with 95% O2-5% CO2. Three minutes following hyperpnea or at the equivalent time, we obtained bronchoalveolar lavage (BAL) and measured its chemiluminescence (CL) counts. In addition, hyperpnea with 95% O2-5% CO2 gas mixture was carried out and BAL was collected 3 minutes after the hyperpnea in an additional forty animals. We measured CL counts in BAL samples before and after the treatments of the following ROS scavenger(s) or saline in vitro: control (saline); superoxide dismutase (SOD); catalase; dimethylthiourea (DMTU); and SOD+catalase+DMTU. Hyperpnea with 95% O2-5% CO2, but not with 95% air-5% CO2, gas mixture induced significant increase in t-butyl hydroperoxide-initiated CL counts, which were inhibited by DMTU, catalase, or SOD in vitro. Our data suggest that hyperpnea with a 95% O2-5% CO2, but not with 95% air-5% CO2, gas mixture induced an increase in ROS production.     

Carbon dioxide as a short-term restraint anaesthetic in rats with subclinical respiratory disease

The use of carbon dioxide (CO2) with, and without, oxygen (O2) as a short-term restraint anaesthetic for Wistar rats in which subclinical respiratory disease was endemic, was assessed in 3 separate experiments. In the first, rats were placed in a CO2 atmosphere generated from solid CO2 chips in a 70 l plastic bin, and removed at time intervals ranging from 0 to 120 s after disappearance of the pedal reflex. Eight of 25 rats died, including 2 which were removed immediately the pedal reflex disappeared; it was concluded that CO2 without O2 was not a suitable short-term anaesthetic for rats. In a second study, rats were anaesthetized in atmospheres of 50:50 and 80:20 (CO2:O2) provided from commercially available cylinders, in 2 different environments--a 3.41 glass jar and a 17 l plastic bin. Rats became excited in the plastic bin but not the glass jar. Rats in the glass jar displayed visible depression and cessation of whiskers movement significantly more quickly in the 80:20 (CO2:O2) than in the 50:50 mixture (4.2 +/- 0.98 s, n = 6, and 66.0 +/- 4.9 s, n = 6 vs 13.8 +/- 2.77 s, n = 5 and 152.0 +/- 20.8 s, n = 5, respectively). Rats in the 17 l plastic bin lost their pedal reflexes in a mean 41.5 +/- 4.55 s (n = 11) in the 50:50 mixture and in a mean 30.9 +/- 6.38 s (n = 11) in the 80:20 (CO2:O2) group. Those left in the 50:50 mixture for 60 s and 180 s after disappearance of their pedal reflexes, recovered these reflexes in 20.2 +/- 0.44 s and 21.5 +/- 7.23 s respectively after removal from the gas. Respiration and heart beat ceased in one rat remaining in the 50:50 mixture after 13 min 10 s. No untoward effects occurred in rats left in the 50:50 mixture for 180 s after disappearance of the pedal reflex, but 2 died when left for an equivalent period in the 80:20 mixture. In the third study, examples of the practical use of a 50:50 mixture as a short term restraint anaesthetic are described. It was concluded that this mixture was a cheap, safe, and effective means of short-term restraint for rats with subclinical respiratory disease, when the minimal time of exposure to the gases was employed.     

Light lamb meat quality packed under modified atmospheres: effect of stunning systems (electrically v. gas)

In the present study, the effect of type of stunning (electrical v. gas) and the gas mixtures in a modified atmosphere (MA) packing system (MA-A: 30% CO2 + 70% O2; MA-B: 30% CO2 + 69.3% N2 + 0.7% CO; MA-C: 40% CO2 + 60% N2) on light lamb meat quality, obtained by 20 males of Manchego Spanish breed, was evaluated at 7, 14 and 21 days post packing. For all ageing times, gas-stunned groups reached the highest pH (P < 0.001) and shear force (SF) values (P < 0.05), and at 21 days, water losses were highest; in contrast, meat colour coordinates were lower (P < 0.001). In general, MA-B showed the highest stability for colour coordinates whereas the lowest tenderness (high SF) was found on MA-A. In this study, the gas mixture with a low carbon monoxide level (MA-B) promoted a higher colour stability and good tenderness, in comparison to the other two blends. In contrast, the conventional packaging system (type A, high oxygen levels) had a reduced tenderness and major colour instability.     

Effects of a local anaesthetic and NSAID in castration of piglets,on the acute pain responses,growth and mortality

The present study addresses the questions whether on-farm use of local anaesthesia with lidocaine leads to a reduction in pain responses during castration, and whether the non-steroidal anti-inflammatory drug meloxicam improves technical performance after castration of piglets. Five treatments were included in the study: (1) castration without anaesthesia or analgesia (CAST), (2) castration after local anaesthesia with lidocaine (LIDO), (3) castration after administration of meloxicam (MELO), (4) castration after lidocaine and meloxicam (L + M) and (5) sham castration (SHAM). To reduce litter influences, each treatment was present in each of the 32 litters (n = 32 per treatment). During castration, vocalizations were recorded continuously. Blood samples were collected 15 min before and 20 min after castration for determination of plasma levels of total cortisol, glucose, lactate and creatine kinase (CK). Mortality was registered and piglets were weighed several times to calculate growth. Several aspects of vocalizations during castration showed consistent and significantly different levels in CAST compared with LIDO, L + M and SHAM. CAST piglets squealed longer, louder and higher. Vocalizations of MELO piglets most resembled those of CAST. An increase in cortisol was seen in all treatments. However, in SHAM piglets this increase was significantly lower than in the other treatments. LIDO piglets showed a significantly smaller increase in plasma cortisol levels compared with CAST and MELO. L + M piglets differed significantly only from the SHAM group. Lactate levels differed significantly between LIDO and MELO, the level in LIDO being decreased after castration. In the other treatments an increase was measured. No treatment effects were found in plasma glucose and CK levels, nor in growth and mortality of the piglets. In conclusion, on the basis of vocalizations and plasma cortisol, local anaesthesia with lidocaine reduces pain responses in piglets during castration. A positive effect of meloxicam on technical performance was not found.     

Physiology and behavior of pigs before and after castration: effects of two topical anesthetics

Surgical castration of male piglets is a common management practice conducted on commercial swine farms to prevent the occurrence of boar taint and aggressive behavior. However, the procedure of surgical castration causes acute pain-induced distress, which is an animal welfare concern. The objective of this study was to evaluate the use of two topical anesthetics to alleviate the pain caused by castration in piglets as measured by physiological and behavior indices of stress. At 3 days of age, 40 weight-matched piglets were allocated to one of four treatment groups. Treatments included: (i) sham castration (CON), (ii) surgical castration (CAS), (iii) castration and short-acting local anesthetic applied topically to the castration wound (SHORT) and (iv) castration and long-acting local anesthetic applied topically to the castration wound (LONG). Blood samples were collected from piglets before and 30, 60, 120 and 180 min after castration to measure leukocyte and differential counts and cortisol concentrations. The above experiment was repeated without blood collection and behavior was recorded for 30 min before and 180 min after castration or handling. Stress vocalizations were recorded during castration and handling. All piglets were weighed before and 24 h after castration and wound healing was recorded daily for the first 14 days after castration. Leukocyte counts and differentials did not differ (P > 0.05) among any of the treatments. Cortisol concentrations were elevated (P < 0.06) in CAS, SHORT and LONG piglets compared with controls 30 and 60 min after castration. The percentage of stress vocalizations was greater (P < 0.05) among castrated piglets compared with CON piglets, regardless of anesthetic treatment. Piglets that were castrated and not given a topical anesthetic spent more time (P < 0.05) lying without contact compared with piglets castrated and given a topical anesthetic, regardless of the topical anesthetic administered. Body weight change did not differ (P > 0.05) among treatments 24 h after castration or control handling and wound healing scores were greater (P < 0.05) in SHORT compared with CAS and LONG piglets 9 to 14 days after castration. In this study, the use of a short- or long-acting topical anesthetic was not effective in reducing the pain-induced distress caused by castration in piglets. Further research is needed to evaluate alternative practical methods to reduce the pain caused by the on-farm castration of piglets.     

Pain behaviour after castration of piglets; effect of pain relief with lidocaine and/or meloxicam

Behavioural responses and the effect of lidocaine and meloxicam on behaviour of piglets after castration were studied. A total of 144 piglets of 2 to 5 days of age were allocated to one of six treatments: castration (CAST), castration with lidocaine (LIDO), castration with meloxicam (MELO), castration with lidocaine and meloxicam (L + M), handling (SHAM) and no handling (NONE). Behaviour was observed for 5 days after the procedure, growth until weaning was recorded and characteristics of the castration wound noted. MELO piglets showed significantly (P < 0.05) more no pain-related behaviour than CAST and LIDO at the afternoon after castration, and were not significantly different from SHAM and NONE. LIDO piglets showed an increase (P < 0.001) in tail wagging, lasting for 3 days. This increase was not seen in L + M piglets. The occurrence of several behaviours changed with age, independent of treatment. A treatment effect on growth was not found. Wound healing was rapid in all treatments, but thickening of the heal was observed in several piglets, suggesting perturbation in the cicatrization process. Our study showed a pain-relieving effect of meloxicam after castration. Local anaesthesia resulted in piglets performing more tail wagging during the first few days after castration, which was prevented by administering meloxicam in combination with local anaesthesia.     

Influence of O(3)/O(2)-pneumoperitoneum as an oxidative stressor on duration of anaesthesia, loss of different reflexes and cytokine mRNA expression

We analysed the effect of intraperitoneal insufflated ozonized oxygen on the anaesthetic strength generated by tribromoethanol, ketamine/xylazine, chloral hydrate, pentobarbital, and urethane in male Wistar rats. High dosages of anaesthetic drugs normally used for deep surgical anaesthesia were injected. The ozonized oxygen gas mixture was given five times daily on five consecutive days at 0.8 mg ozone/kg body weight before anaesthesia. The reflexes were measured 15, 30, 60, 90, 120, 180, and 240 min after injection of the anaesthetic drug. The sleeping time and the loss and regain of six different reflexes on noxious and non-aversive stimuli were recorded during the 4 h of observation. O(3)/O(2)-pneumoperitoneum (O(3)/O(2)-PP) reduced the sleeping time induced by tribromoethanol and ketamine/xylazine and increased it for chloral hydrate and pentobarbital. In accordance to the changes in the duration of anaesthesia, the O(3)/O(2)-PP induced significant changes in the loss of different reflexes. Additionally, the modulatory effect of the anaesthetic drugs on splenic cytokine mRNA expression was further influenced by O(3)/O(2)-PP. Thus, the influence of an oxidative stressor on anaesthetic potency and on the resting immune system has to be taken into account for experimental designs in which surgical anaesthesia is necessary for small laboratory animals.     

Effects of hyperoxia on vasoconstriction and VA/Q matching in the neonatal lung

Exposure of adult animals to 48-72 h of 100% O2 breathing is associated with a blunting of hypoxic pulmonary vasoconstriction (HPV) (Newman et al. J. Appl. Physiol. 54: 1379-1386, 1983). It is unknown whether HPV is also diminished in neonates after hyperoxic exposure and if so to what extent such suppression might interfere with pulmonary gas exchange during hypoxic gas breathing. We tested the possibility that hyperoxia would suppress HPV and interfere with ventilation-perfusion (VA/Q) matching and therefore gas exchange in neonatal piglets. Twelve 2- to 4-wk-old piglets were exposed for an average of 68 h to greater than 90% inspired O2. A control group of eight piglets was exposed to room air for a similar period of time. Immediately after exposure the animals were anesthetized and instrumented. Pulmonary hemodynamics and respiratory and inert gas exchange were assessed while the animals inspired an O2 fraction of 1.0, 0.21, and 0.12. After 20 min of hypoxic gas breathing, pulmonary arterial pressure rose to a lesser degree in the hyperoxia (H)-exposed animals than in the control (C) animals (P less than 0.02). The increase in pulmonary vascular resistance was similarly blunted. Venous admixture of the insoluble inert gas, sulfur hexafluoride, an index of extremely low VA/Q areas, was increased during hypoxic gas breathing compared with room air breathing in the H-preexposed animals (P less than 0.02). Standard deviation of pulmonary blood flow was increased (P less than 0.02), indicating an increase in mismatching of VA/Q during hypoxic breathing in the H-preexposed animals compared with the C animals.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pulmonary and bronchial blood circulation under conditions of changed gaseous environment

By means of ultrasonic method used in acute experiments on cats with closed chest under normal respiration the authors studied the blood flow in left low-lobar pulmonary artery and vein and in bronchial artery, as well as the blood pressure in pulmonary and femoral arteries in inhalation of next gaseous mixtures: 7.5% O2 in nitrogen; 30% O2; 3% CO2; 21% O2+ +79% He; 30% O2 + 67% He + 3% CO2. It was shown, that inhalation of the normoxic gaseous mixture, in which nitrogen is replaced by helium, did not call significant changes in pulmonary and systemic circulation. However, the presence of the helium in complicated gaseous mixture can change the reactivity of pulmonary and bronchial vessels to influence the components participating in these complicated gaseous mixtures.     

Assessing recruitment of lung diffusing capacity in exercising guinea pigs with a rebreathing technique.

Noninvasive techniques for assessing cardiopulmonary function in small animals are limited. We previously developed a rebreathing technique for measuring lung volume, pulmonary blood flow, diffusing capacity for carbon monoxide (Dl(CO)) and its components, membrane diffusing capacity (Dm(CO)) and pulmonary capillary blood volume (Vc), and septal volume, in conscious nonsedated guinea pigs at rest. Now we have extended this technique to study guinea pigs during voluntary treadmill exercise with a sealed respiratory mask attached to a body vest and a test gas mixture containing 0.5% SF(6) or Ne, 0.3% CO, and 0.8% C(2)H(2) in 40% or 98% O(2). From rest to exercise, O(2) uptake increased from 12.7 to 25.5 ml x min(-1) x kg(-1) while pulmonary blood flow increased from 123 to 239 ml/kg. The measured Dl(CO), Dm(CO), and Vc increased linearly with respect to pulmonary blood flow as expected from alveolar microvascular recruitment; body mass-specific relationships were consistent with those in healthy human subjects and dogs studied with a similar technique. The results show that 1) cardiopulmonary interactions from rest to exercise can be measured noninvasively in guinea pigs, 2) guinea pigs exhibit patterns of exercise response and alveolar microvascular recruitment similar to those of larger species, and 3) the rebreathing technique is widely applicable to human ( approximately 70 kg), dog (20-30 kg), and guinea pig (1-1.5 kg). In theory, this technique can be extended to even smaller animals provided that species-specific technical hurdles can be overcome.     

Milk intake and carbon dioxide production of piglets determined with the doubly labelled water technique

The present study was undertaken to study different methodological aspects of quantifying CO2 production and milk intake of suckling piglets using the doubly labelled water (DLW) technique. In total, 37 piglets were enriched intraperitoneally with DLW to study equilibration time of 18O (n = 3), to validate the estimation of milk intake and CO2 production (n = 10) of piglets fed milk replacer and to quantify milk intake and CO2 production of piglets nursed ordinarily by sows (n = 24). Enrichment of 18O in expired air was analysed without any sample preparation, whereas enrichment of 18O in serum was analysed after a minimum step of sample preparation, which included pipetting of the sample, blowing gaseous CO2 into the vial for 3 s and equilibrating for 24 h. The 18O enrichment of CO2 in expired air was constant within 30-40 min of intraperitoneal injection, suggesting that DLW was equilibrated within the body water by that time. For piglets fed milk replacer, the estimation of the daily CO2 production by the DLW method (64.0 ± 2.7 l CO2/day) was in agreement with that obtained by respiration trials (64.7 ± 1.8 l CO2/day). Furthermore, the intake of milk replacer (891 ± 63 g/day) determined by deuterium oxide (D2O) dilution was similar in magnitude to that found by weighing the milk disappearance (910 ± 58 g/day). The milk intake of piglets fed milk replacer was comparable with that of sucking piglets, but sucking piglets had a remarkably higher CO2 production than artificially reared piglets, which likely was caused by a higher intake of milk solids and a higher activity level. For sucking piglets, the daily CO2 production increased curvilinearly with increasing live weight (LW) in kg: piglet CO2 production (l/day) = 25.75 × LW - 1.01 × LW2. In conclusion, 18O equilibrates fast within the body water pool when administered intraperitoneally, and the accuracy of assessing milk intake and rate of CO2 production using the DLW technique is promising. Assessment of excess enrichment of 18O in serum proved to be robust. Finally, the CO2 production of piglets fed milk replacer differs considerably from that of sucking piglets.     

Behavioural responses of piglets to castration: the effect of piglet age

It has long been assumed that neonatal animals are less sensitive than older animals to pain, and this reasoning has been used to recommend that routine surgical procedures be performed at an early age. In this study we tested if vocal and other behavioural responses to castration increase with piglet age. Piglets (n=84) from 14 litters were assigned to one of six treatment groups: castration or sham castration at 3, 10 or 17 days of age. During the procedure castrated piglets produced high-frequency calls (>1000Hz) at more than three times the rate of piglets in the sham-castrate group. The rate of low-frequency (<1000Hz) calls was also higher for piglets in the castrate group. The rate of high-frequency calling was lower for the youngest pigs but there was no relationship between age and the effect of treatment for any of the vocal responses measured (i.e. no age by treatment interaction). During the first 2h after castration, castrated piglets spent more time sitting or standing and less time lying. During the subsequent 22h, castrated piglets spent marginally more time at the udder and less time lying down. Older piglets missed more nursings. However, the effect of castration did not vary with the age of the piglet for any measure. We conclude that while the factors affecting both the shams and the castrates (e.g. distress due to restraint) may vary with age, the pain of castration is not affected by age within the range of ages that we tested.     

Monoamine Synthesis and Concentration in Neonatal Rat Brain During Hypercapnia and Recovery

One-day-old rats were exposed to a gas mixture of 15% CO2-21% O2-64% N2 for a 30-min period. Monoamine synthesis in whole brain was measured during, and at various intervals after, hypercapnia by estimating the accumulation of dihydroxyphenylalanine (DOPA) and 5-hydroxytryptophan (5-HTP) after inhibition of aromatic L-amino-acid decarboxylase with NSD 1015. Endogenous concentrations of tyrosine, dopamine (DA), noradrenaline (NA), tryptophan, 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were measured at the same intervals. Exposure to CO2 induced an increased synthesis of catecholamines and 5-HT. Further, an increase in DA concentration was seen during hypercapnia, while NA and 5-HT were unchanged. After the CO2 exposure the increased in vivo synthesis rates of catecholamines and 5-HT were rapidly normalized, as was the endogenous DA concentration. A slight increase in 5-HT and 5-HIAA concentrations was seen immediately after CO2 exposure. These results indicate that in neonatal animals, hypercapnia induces changes in central monoamine neurons, primarily an increased synthesis. These alterations may be relevant to some physiological changes seen during CO2 exposure, such as the alteration in central respiratory performance.     

Castration inhibits exercise-induced accumulation of Hsp70 in male rodent hearts

Intense exercise leads to accumulation of the inducible member of the 70-kDa family of heat shock proteins, Hsp70, in male, but not female, hearts. Estrogen is at least partially responsible for this difference. Because androgen receptors are expressed in the heart and castration leads to decreases in calcium regulatory proteins and altered cardiac function, testosterone (T) or its metabolites could also be involved. We hypothesized that removal of endogenous T production through castration would reduce cardiac Hsp70 accumulation after an acute exercise bout, whereas castrated animals supplemented with 5alpha-dihydrotestosterone (DHT) would show the intact male response. Fifty-four 8-wk-old male Sprague-Dawley rats were divided into intact, castrated, or castrated + DHT groups (n = 18/group). At 11 wk of age, 12 animals in each group undertook a 60-min bout of treadmill running at 30 m/min (2% incline) while the remaining 6 in each group remained sedentary. At 30 min or 24 h after exercise (n = 6/time point), blood and hearts were harvested for analysis. Serum T was undetectable in castrated and DHT-treated castrated rats, whereas serum DHT was significantly reduced in castrated animals only (approximately 60% reduction) (P < 0.05). Although there were no differences in constitutive levels of Hsp70 protein, exercise significantly increased cardiac hsp70 mRNA and protein in intact and DHT-supplemented rats, but not in castrated animals (P < 0.05). To examine whether castration eliminated the ability to respond to stress, another six intact and six castrated animals were subjected to a 15-min period of hyperthermia (core temperature raised to 42 degrees C) and killed 24 h later. As opposed to exercise, castrated animals subjected to heat shock exhibited increases in Hsp70 above nonshocked (i.e., sedentary) animals, similarly to intact males (P < 0.05). These data suggest that androgens, in addition to estrogen, play a role in the sexual dimorphism observed in the stress response to exercise but not heat shock.     

Tolerance of Low Intracellular pH During Hypercapnia by Rat Cortical Brain Slices: A 31P/1H NMR Study

Metabolic tolerance of low intracellular pH (pH(i)) was studied in well-oxygenated, perfused, neonatal, rat cerebrocortical brain slices (350 microns thick) by inducing severe hypercapnia. In each of 17 separate experiments 80 brain slices (approximately 3.2 g wet weight) were suspended in an NMR tube, perfused with artificial CSF (ACSF), and studied at 4.7 T with 31P and 1H NMR spectroscopy. Spectra obtained every 5 min monitored relative concentrations of lactate or high-energy phosphate metabolites, from which pH(i) and extracellular pH were determined. Unperturbed slice preparations were metabolically stable for > 10 h, with no significant changes occurring in pHi, ATP, phosphocreatine (PCr), inorganic phosphate, or lactate. Different levels of hypercapnia were produced by sequentially perfusing slices with the following different ACSF batches, each having previously been equilibrated with a specific mixture of CO2 in oxygen: (a) 10% CO2, 15 min of perfusion; (b) 30% CO2, 15 min of perfusion; (c) 50% CO2, 15 min of perfusion; (d) 70% CO2, 30 min of perfusion; (e) 50% CO2, 15 min of perfusion; (f) 30% CO2, 15 min of perfusion; and (g) 10% CO2, 15 min of perfusion. At the completion of this protocol slices were again perfused with fresh ACSF that was equilibrated with a 95% O2/5% CO2 gas mixture. In each of five separate 1H and 31P experiments, brain slices were recovered within 2 h after termination of exposure to high CO2. The pHi was determined from measurements of the chemical shift difference between phosphoethanolamine and PCr, using a calibration curve obtained for our preparation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Impact of general injection anaesthesia and analgesia on post-castration behaviour and teat order of piglets

Injection anaesthesia with a combination of ketamine and azaperone (K/A) is discussed as a painless alternative to commonly used non-anaesthetized castration. To protect anaesthetized piglets from being crushed, they have to be separated from the sow for 3 h following castration. The aim of this study was to test if this separation and the different treatments would affect short-term behaviour after castration (3 to 6 h after castration) as well as weight gain. Piglets were 5 to 7 days old. Treatment Group 1 received a combination of anaesthesia and analgesia (n = 29, ketamine: 25 mg/kg BW; azaperone: 2 mg/kg BW; meloxicam: 0.4 mg/kg BW), Group 2 received only analgesia (n = 24) and Group 3 received no medication (n = 29). Behaviour and suckling order were compared for a 3 h period the day before castration and after castration. A significantly higher number of teats used by anaesthetized piglets (P = 0.004) suggests a decrease in suckling order stability. There were significant treatment effects between all three groups in the time spent at the sow's teat, with an increase in Group 2 (+69%), decrease in Group 1 (−28%), whereas the control Group 3 (+2%) almost remained unchanged. The anaesthetized piglets showed an increase in the time spent active away from the sow after castration of almost 200% (Groups 2 and 3: ∼50%, P < 0.001). However, no significant treatment effect was seen for weight gain. The results suggest that analgesia has an effect on behaviour, perhaps due to less post-castration pain. This advantage is not apparent for animals receiving additional anaesthesia, probably because of impaired coordination. Although the behavioural changes did not affect weight gain significantly, a decrease in suckling order stability indicates a certain degree of stress due to fighting over teat positions as a consequence of separation. Thus, post-castration behaviour must be taken into account when evaluating alternative castration methods.     

Gas exchange abnormalities after pneumonectomy in conditioned foxhounds

Loss of a major portion of lung tissue has been associated with impaired exercise capacity, but the underlying mechanisms are not well defined. We studied the alterations in gas exchange during exercise before and after left pneumonectomy in three conditioned foxhounds. After pneumonectomy, minute ventilation and O2 consumption at comparable submaximal work loads were unchanged but arterial PCO2 at any work load was higher, implying that ventilatory response to CO2 was impaired. Arterial hypoxemia and an elevated alveolar-arterial O2 tension difference (AaDO2) developed during heavy exercise. Using the multiple inert gas elimination technique, we determined the distributions of ventilation-perfusion (VA/Q) ratios postpneumonectomy. Significant increase in VA/Q inequality developed during exercise while the foxhounds were breathing room air, accounting for an average of 42% of the total increase in AaDO2 while diffusion limitation accounted for 58%. While the animals were breathing hypoxic gas mixture, diffusion limitation accounted for an average of 88% of the total increase AaDO2. Cardiac output and O2 delivery were reduced at a given O2 consumption after pneumonectomy. After pneumonectomy, the animals reached O2 consumptions close to the maximum expected for normal dogs. Compensation for the impairment in O2 delivery post-pneumonectomy occurred mainly by an increase in hemoglobin concentration. Training probably played an important role in returning exercise capacity toward prepneumonectomy levels. We conclude that significant abnormalities in gas exchange develop during exercise after loss of 42% of lung tissue, but the animals demonstrate a remarkable ability to compensate for these changes.     

Hypoxic pulmonary vasoconstriction does not affect hydrostatic pulmonary edema formation

We studied the effects of regional hypoxic pulmonary vasoconstriction (HPV) on lobar flow diversion in the presence of hydrostatic pulmonary edema. Ten anesthetized dogs with the left lower lobe (LLL) suspended in a net for continuous weighing were ventilated with a bronchial divider so the LLL could be ventilated with either 100% O2 or a hypoxic gas mixture (90% N2-5% CO2-5% O2). A balloon was inflated in the left atrium until hydrostatic pulmonary edema occurred, as evidenced by a continuous increase in LLL weight. Left lower lobe flow (QLLL) was measured by electromagnetic flow meter and cardiac output (QT) by thermal dilution. At a left atrial pressure of 30 +/- 5 mmHg, ventilation of the LLL with the hypoxic gas mixture caused QLLL/QT to decrease from 17 +/- 4 to 11 +/- 3% (P less than 0.05), pulmonary arterial pressure to increase from 35 +/- 5 to 37 +/- 6 mmHg (P less than 0.05), and no significant change in rate of LLL weight gain. Gravimetric confirmation of our results was provided by experiments in four animals where the LLL was ventilated with an hypoxic gas mixture for 2 h while the right lung was ventilated with 100% O2. In these animals there was no difference in bloodless lung water between the LLL and right lower lobe. We conclude that in the presence of left atrial pressures high enough to cause hydrostatic pulmonary edema, HPV causes significant flow diversion from an hypoxic lobe but the decrease in flow does not affect edema formation.