Haemolytic activity of sheep complement for two assay systems

Sheep complement (C) is haemolytic for sheep erythrocytes sensitized with rabbit antibody (sheep E-rabbit A) provided serum is used as soon as possible after collection. If left at 4 °C to separate from the clot, serum C activity for sheep E-rabbit A is markedly reduced. Heparinized plasma retains its haemolytic titre for at least 24 h at 4 °C. Plasma from Mg²⁺-ethyleneglycoltetraacetic acid (EGTA) blood is non-haemolytic, but addition of Ca²⁺ partially restores the titre. A high concentration of rabbit A is necessary to sensitize sheep E.Sheep C is haemolytic for human erythrocytes sensitized with sheep antibody (human E-sheep A) in the presence of Mg²⁺-EGTA. This C activity is stable at 4 °C for 24 h in serum, Mg²⁺-EGTA plasma and heparinized plasma. Haemolytic activity of serum heated at 50 °C for 30 min was restored by a factor B containing CM-cellulose fraction of foetal lamb serum in the presence of Mg²⁺-EGTA for human E-sheep A but not sheep E-rabbit A.These findings show that sheep C haemolysis of sheep E-rabbit A requires a Ca²⁺-Mg²⁺-dependent pathway that is labile in vitro for 24 h at 4 °C.     

Anti-complementary activity for guinea-pig and sheep complement in serum and Mg2+-EGTA plasma from various animals

Serum and Mg²⁺-ethyleneglycoltetraacetic acid (EGTA) plasma from 14 domestic or laboratory animal species had no anti-complementary activity for sheep complement (C). Serum samples from 8 species and all 14 Mg²⁺-EGTA plasma samples were anti-complementary towards guinea-pig C. Thus, use of sheep C in a warm complement fixation text (with human erythrocytes sensitized with sheep antiserum, IgM, slow or fast γ-globulin antibody), in the presence or absence of Mg²⁺-EGTA, may be a satisfactory method of eliminating anticomplementary activity.     

The effects of Leptospira serotype pomona in sheep of different haemoglobin types

A warm complement fixation test that will detect antibody in sheepserum in the virtual absence of anti-complementary activity is described. Sheep antibody-antigen complexes were detected by fixation of sheep complement. Sheep serum, heparinized or Mg⁺⁺—EGTA plasma was used as the source of sheep complement. Sheep-antibody-sensitized human erythrocytes were used as the haemolytic indicator cells for sheep complement. As the modified complement-fixation test was performed in the presence of Mg⁺⁺—EGTA, sheep C probably reacts with sheep Ab-Ag complexes by a different mechanism than does guinea-pig complement.     

Variations in sheep serum conglutinating and haemolytic activity for sheep erythrocytes sensitized by rabbit antibody

Based on conglutinating and haemolytic reactions with sheep erythrocytes (E) sensitized by rabbit antibody (A), three types of sheep sera were encountered. Type 1 sera do not conglutinate or haemolyse sheep E-rabbit A. Type 2 sera failed to conglutinate, but are haemolytically active. Type 3 sera have both activities. Serum from one type 1 sheep still failed to conglutinate 5 days after venepuncture but was now haemolytically active (i.e., type 2). Some sheep that initially had type 2 sera had, five days after an intraperitoneal injection of yeast cells, sera with conglutinating activity (type 3 sera). Type 1, 2 and 3 sera all had haemolytic activity with human E-sheep A indicator cells.Pooled type 3 sera have the highest conglutinating titres with sheep E-rabbit A after 10 min incubation at 39 °C. At this stage, the haemolytic titres were very low. From 10 min, the conglutinating titres decreased whereas the haemolytic titres gradually increased until 80 min. Optimal conglutinating activity required less rabbit A to sensitize sheep E than did haemolytic activity.     

Demonstration of Alternative and Classical Complement Pathway Activity in Colostrum from Buffalo (Bubalus bubalis)

Buffalo colostrum caused lysis of unsensitized red blood cells (RBC) from sheep, goats, rabbits and chickens. RBC from cattle and buffalo were resistant to lysis. That lysis was due to the presence of natural antibodies to these RBC was ruled out since there was no reduction in haemolytic titres even after adsorption with the respective RBC. The addition of EGTA to the diluent had no effect on the haemolytic activity. These findings indicate the presence of alternative complement pathway (ACP) activity in buffalo colostrum. The haemolytic activity of buffalo complement for unsensitized rabbit RBC was reduced to very low levels by heating at 50°C for 45 min. Treatment with zymosan also inhibited the haemolytic activity, while inulin had no effect. The maximum activity of ACP occurred in the presence of 4 mmol/L Mg²⁺ in the diluent. The range of ACP activities in colostrum from buffaloes varied from 4.06 to 8.48 CH₅₀ units/ml. Using a standard system for titrating the classical complement pathway and rabbit red blood cells sensitized with goat haemolysin, the range of complement activity in buffalo colostrum was 4.81–6.77 CH₅₀/ml.     

Identification and characterization of serum complement activity in the American crocodile (Crocodylus acutus)

Incubation of unsensitized sheep red blood cells with serum from the American crocodile (Crocodylus acutus) resulted in a concentration-dependent hemolysis. The hemolytic activity was heat-sensitive, and inhibited by EDTA in a concentration-dependent manner. The EDTA-inhibited SRBC hemolysis could be restored by the addition of excess Ca²⁺ or Mg²⁺, but not Ba²⁺ or Cu²⁺, revealing the specificity of this activity for these two divalent cations. The hemolytic activity of crocodile serum was titer-dependent, with 329 μL producing 50% of maximal SRBC hemolysis. The complement activity was also temperature-dependent, with decreased activity at lower temperatures (5–15 °C) and maximal activity occurred at 30–40 °C. The hemolysis occurred relatively slowly, with near zero activity after 10 min, 40% of activity observed within 15 min of exposure to SRBCs, and maximal activity at 30 min.     

Defences of the bovine mammary gland against infection and prospects for their enhancement

The equine alternative complement pathway has been partially characterized and compared to the equine classical activation pathway. A dose-dependent lysis of RbRBC was observed with peak lytic values noted within 10 minutes at 37°C when rabbit red blood cells (RbRBC) were used as an alternative pathway activator. Sheep red blood cells (SRBC) sensitized with rabbit hemolysin or partially purified equine IgM antibodies were equally sensitive to lysis. Dilution of the commercial hemolysin by $\text{1}\text{5}$ reduced lysis from 90% to 38% in the presence of constant cell numbers. Hemolysis of SRBC peaked at 10 minutes and the majority of lysis occurred within 10 minutes. Dilution of equine sera by as little as $\text{1}\text{5}$ decreased hemolytic activity for SRBC to 21.5% from greater than 90% with undiluted sera. The alternative pathway protein, equine factor B, was tested using RbRBC and monitored by its differential susceptibility to heat treatment at 50°C. This treatment led to almost complete inactivation after a 15-minute incubation. An apparent heat-dependent decay of certain classical pathway components was also observed after 50°C treatment. This sensitivity was indicated by a reduction in the lytic activity for sensitized SRBC. Treatment for 15 minutes at 56°C with either RbRBC or SRBC was sufficient to abolish hemolytic activity in all equine sera tested. Chelation of cations with 0.04 M EDTA blocked expression of alternative and classical pathway activation; however, chelation of Ca⁺⁺ ions with 10 mM EGTA containing 1 mM Mg⁺⁺ ions permitted lysis of the RbRBC but not the SRBC. A dose-related Mg⁺⁺-ion dependence for RbRBC hemolytic activity was observed as the concentration of Mg⁺⁺ was increased to 1.0 mM. In addition, our results obtained with pre-colostral foal serum strongly suggest that natural antibody to RbRBC was of little importance in the lysis observed with these cells. These results also show that the equine alternative pathway activation may require Ca⁺⁺ ions. If Ca⁺⁺ ions are required, the equine alternative pathway is quite different from any other mammalian complement system so far described. Our results suggest that the alternative pathway of activation is of major importance in the equine complement system. Confirmation of this hypothesis requires both purification of the components involved as well as further characterization.     

Studies on activation and levels of haemolytic complement of buffalo (Bubalus bubalis). II. Alternate complement pathway activity in serum.

Buffalo serum caused lysis of unsensitized red blood cells (RBC) of sheep, goat, rabbit and guineapig. There was minimal lysis of cattle RBC, and homologous RBC were resistant. Lysis of sheep and goat RBC was the result of natural antibodies as adsorption with respective RBC and addition of 8 mmol ethylene glycolbistetraacetate (EGTA) in diluent completely abrogated the haemolytic activity. The lysis of guinea-pig and rabbit RBC was only partially decreased by these treatments, indicating the presence of alternate complement pathway (ACP) activity in buffalo serum. The guinea-pig RBC were the most sensitive to lysis, and 50% CH titre units above 40 ml⁻¹ of serum were obtained. The haemolytic activity of buffalo C for unsensitized guinea-pig RBC was reduced from 47 CH₅₀ units to an undetectable level by heating at 50°C for 20 min and at 56°C for 4 min. Similarly, treatment with zymosan also inhibited this haemolytic activity. Maximum activation of buffalo ACP occurred in the presence of 4 mmol Mg²⁺ in the diluent.

Using standardized conditions, ACP activity was determined in sera of 98 healthy buffaloes of different age groups from 1 month to 12 years. Even young calves less then three months of age showed considerable ACP activity (45.60±1.21 CH₅₀ units ml⁻¹) which increased with age. The peak mean values of 79.79±1.45 CH₅₀ units was recorded in 2 to 4-year-old animals. However, in all the 11 animals above 4 years of age, the haemolytic activity was greatly reduced and was even less than that in 1 to 3-month-old buffalo calves. Haemolytic activity did not vary between the sexes.     

Serum complement activity in the three-toed amphiuma (Amphiuma tridactylum)

Some animals routinely endure serious injuries from predators or during intraspecific territorial conflicts. Such is the case for Amphiuma tridactylum, an aquatic salamander that lives in an environment rich in potentially infectious microbes, apparently with rare or no pathogenic infection. Some vertebrates possess innate immune mechanisms, but whether this is the case for Amphiuma is unknown. To assess this potential, plasma from 19 A. tridactylum was pooled and used for characterisation of serum complement activity. The ability of A. tridactylum plasma to hemolyse unsensitised sheep red blood cells (SRBCs) was titer-dependent, with low activity observed even at high plasma titers. The kinetic characterisation of SRBC hemolysis revealed that significant activity could be measured within 10 min of incubation, and maximal activity occurred within 60 min. The SRBC hemolysis by A. tridactylum plasma was also temperature-dependent, with maximal activity at 30 °C. In addition, this activity was sensitive to mild heat treatment, with 96% of activity inhibited by incubation at 56 °C for 30 min. The SRBC hemolysis could also be inactivated by pretreatment of the plasma with proteases, indicating that this activity was protein dependent. The activity required divalent metals ions, with activity inhibited by EDTA, citrate, or phosphate. However, the chelator-inhibited activity could be restored by the addition of excess Ca²⁺ or Mg²⁺, but not Cu²⁺ or Ba²⁺, indicating specificity of the divalent metal ion requirement. The sensitivity to heat, proteases, and divalent metal ion chelators strongly suggests that A. tridactylum plasma-mediated hemolysis of SRBCs is mediated by the serum complement system of proteins.     

Molecular Size,pH, Temperature Stability,and Ontogeny of Inhibitor(s) of Infectious Pancreatic Necrosis Virus (IPNV) in Normal Rainbow Trout Serum

Abstract

In this study, we investigated the characteristics of inhibitor(s) of infectious pancreatic necrosis virus (IPNV) found in the serum of normal rainbow trout Oncorhynchus mykiss (RTS). The molecular size, stability to pH and temperature, and ontogeny of the inhibitor in trout were studied, and the effect of cations (Ca²⁺ and Mg²⁺) on the activity of the inhibitor was tested. The strongest inhibition of virus was obtained at approximately 150 kDa as measured by ultracentrifugation, sieve gel chromatography, and ultrafiltration. The inhibition decreased significantly when RTS was dialyzed or filtered in the absence of divalent cations, but replacement of at least one cation restored activity. Activity was stable at temperatures ranging from 30°C to 50°C, but 55°C completely destroyed the inhibitory capacity of RTS. The inhibitory activity of RTS was not reduced between pH 4 and 10 but was diminished below pH 4 and above pH 10; such activity was not abrogated by proteases. Additionally, pretreatment of RTS with the polysaccharide mannan significantly reduced inhibition. Thus, the serum inhibitor(s) had many characteristics of a lectin. To determine the ontogeny of inhibition, serum samples were taken from normal rainbow trout, beginning at 2 weeks posthatch; consistent inhibition was not obtained until the rainbow trout had reached the age of 23 weeks posthatch.     

Cryopreservation of sheep red blood cells. 3. Complement titrations with frozen sensitized cells

The effect of different suspending and washing procedures for recovery of sensitized sheep erythrocytes (EA) after freezing at −196°C was investigated. Best results were obtained using gelatin-veronal-buffered saline-sucrose containing 0.15 mM-Ca and 1 mM-Mg (GVBSM⁺⁺-sucrose) as the suspending and first washing buffer. The cryoprotective agents tested were polyvinylpyrrolidone (PVP), neutralized PVP, purified PVP and a gum product, Avelex 1030. All PVP preparations tested gave good results as cryoprotectants in terms of cell recovery after thawing whereas Avelex 1030 was less satisfactory. The EA cells frozen in the presence of untreated PVP showed, however, increased susceptibility to the hemolytic action of complement, whereas cells frozen with purified or neutralized PVP gave titers similar to that obtained with fresh cells. Good results were also obtained with Avelex 1030. Complement titrations with frozen EA cells were more reproducible than titrations with fresh cells.     

Proteolytic system of Bacillus sp. CL18 is capable of extensive feather degradation and hydrolysis of diverse protein substrates

1. Feathers are recalcitrant protein-rich wastes produced in huge amounts by poultry processing for meat production. Hence, feather bioconversion and protease production by Bacillus sp. CL18 were investigated.

2. Bacillus sp. CL18 demonstrated a remarkable feather-degrading potential. Through cultivations on feather broth (10 g l^?1 feathers), 94.5% ± 3% of whole feathers were degraded after 4 d. Increases in soluble protein contents were observed and protease production was maximal also at d 4. This strain produced diverse proteolytic enzymes during growth.

3. Crude protease displayed optimal activity at 55°C (50–62°C), pH 8.0 (7.0–9.0) and a low thermal stability. Proteolytic activity increased in the presence of Ca²⁺, Mg²⁺, Triton X-100, Tween 20 and dimethyl sulphoxide. Inhibition profile indicated that crude protease contains, mainly, serine proteases. Enzyme preparation hydrolysed mainly casein and soy protein isolate.

4. The keratinolytic capacity of Bacillus sp. CL18 at moderate temperatures (30°C) might be appropriate for feather conversion, resulting in protein hydrolysates and proteolytic enzymes. Proteases are postulated to be added-value products that can be obtained from such a bioprocess.     

Erythrocyte metabolism in normal and glutathione-deficient sheep.

Glucose utilization and lactate formation in erythrocytes from normal and glutathione (GSH)-deficient sheep were similar. Significant differences were observed, however, between the 2 groups of sheep in the production of 14-CO2 from erythrocytes incubated with ascorbic acid or methylene blue, or both. The greater response of normal erythrocytes compared to erythrocytes deficient in GSH suggests that there are some metabolic differences in the pentose phosphate pathway (ppp) activity of the erythrocytes. The nature and site of these differences are, however, not known. When sheep were kept in a decompression chamber for 2 weeks and subjected to a simulated altitude of 7,000 m for 12 hours/day, the erythrocytes showed a four- to six-fold increase in the activity of the ppp.     

Characterization of lymphocyte subpopulations in sheep by rosette formation,adherence to nylon wool and mitogen responsiveness

Sheep peripheral blood lymphocytes have been studied using a number of surface markers. Thus 16.6 ± 2.4% (mean ± S.E.) were surface immunoglobulin positive (sIg⁺) by direct immunofluorescence, 35.9 ± 2.1% formed Fc rosettes with bovine red blood cells (RBC) sensitized with rabbit antibody (Fc⁺) and 28.4 ± 2.0% formed rosettes with sheep red blood cells (RBC) in the presence of 4% dextran (DS⁺). The percentage of both Fc⁺ and DS⁺ lymphocytes tended to increase with age of the animals. Demonstration of these markers allowed computation of two further subpopulations: null cells lacking sIg and a receptor for sheep RBC, and Fc·null cells lacking a receptor for Fc and sheep RBC. The former population, which contained a proportion of Fc⁺ lymphocytes comprised 49.8 ± 3.8% of blood lymphocytes and the latter 38.4 ± 3.0%.Separation on nylon wool columns, selective rosette enrichment and depletion on density gradients and stimulation with phytomitogens have shown sIg⁺ and Fc⁺ lymphocytes to be nylon wool adherent and unresponsive to phytohaemagglutinin (PHA) and Concanavalin A (Con A) and DS⁺ lymphocytes to be nylon wool non-adherent and responsive to PHA and Con A. The data also indicates a major overlap of the lymphocyte subpopulations bearing sIg and Fc which are apparently B lymphocytes. Moreover these data support the contention that E-rosette formation with sheep RBC in the presence of dextran is a marker for sheep T cells. The data also indicates that Fc·null cells are T cells, eluting in the non-adherent fraction from nylon wool. It is probable that a proportion of these cells bear a SRBC receptor too weak for present detection methods.     

Differences in body temperature, cell viability, and HSP-70 concentrations between Pelibuey and Suffolk sheep under heat stress

Pelibuey and Suffolk sheep were compared as to their capacity to regulate body temperature under environmental hyperthermia by measuring their differences in cellular response to heat stress (HS). In a first experiment, seven Pelibuey and seven Suffolk ewes were kept in a climatic chamber for 6 h daily during 10 days (temperatures within the 18 to 39.5 °C range). As chamber temperature rose, sheep rectal temperature increased in both groups, but to a lesser extent in Pelibuey (0.3 °C) than in Suffolk sheep (0.7 °C) (P?<?0.05). In a second experiment, cellular viability was assessed using cultured blood mononuclear cells from 15 Pelibuey and 15 Suffolk sheep. They were incubated at 37 °C for 24 h (control) or 43 °C for 6 h followed by 18 h at 37 °C (HS). In a third experiment, another blood mononuclear cells culture from eight Pelibuey and eight Suffolk sheep was kept at 37 °C for 15 h; these were subsequently cultured for 6 h at 37 °C (controls) or 43 °C (HS). Next, HSP-70 concentration was determined. HS reduced the percentage of viable cells to a greater extent in Suffolk [37 °C (73.7 %) vs. 43 °C (61.9 %); P?<?0.05] than in Pelibuey sheep [37 °C (74.9 %) vs. 43 °C (66.7 %); P?>?0.05]. HS significantly increased HSP-70 average concentrations for both breeds at 43 °C. A significant effect was observed for the breed by temperature interaction (P?<?0.05) caused by a greater difference between Pelibuey and Suffolk at 43 °C (2.85 vs. 0.53 ng/mL, respectively; P?<?0.05) than at 37 °C (0.05 vs. 0.03 ng/mL, respectively; P?>?0.05). In conclusion, Pelibuey sheep show more effective body temperature regulation under conditions of environmental hyperthermia. Also, cell viability after HS was higher in Pelibuey than in Suffolk, an effect that could be mediated by an HSP-70-related mechanism.     

Antibody-dependent cell-mediated cytotoxicity in sheep

The ability of sheep leukocytes to mediate antibody — dependent cell-mediated cytotoxicity (ADCC) and that of sheep serum IgG₁ and IgG₂ to induce ADCC were investigated. Partial characterization of effector cells was attempted. These investigations revealed that ADCC occurs in sneep. With chicken erythrocytes (CRBC) as the target cells, polymorphonu-cleated cells (PMN), and monocytes, were the most effective leukocytes. Ovine peripheral blood lymphocytes (PBL) also mediated ADCC, and within the PBL population, T-cells were capable of mediating ADCC. The T-cells were obtained by nylon wool fractionation and selective agglutination by peanut agglutinin (PNA) and Helix pomatia agglutinin (HPA). Both nylon wool adherent and non-adherent fractions were active in ADCC, although the former were more active, implying heterogeneity in nylon wool adherence among ovine K-cells. Depletion of B (SIg⁺) cells did not affect ADCC activity of the remaining cells. Depletion of Fc⁺ cells markedly reduced cytotoxic activity of PBL. Both sheep IgG₁ and IgG₂ anti-CRBC immunoglobulins were able to induce ADCC.     

Thermal and mechanical nociceptive threshold testing in pregnant sheep

ObjectiveAnalgesic regimes were compared in pregnant ewes after laparotomy by measuring thermal (TT) and mechanical (MT) nociceptive thresholds.Study designProspective randomised experimental study.AnimalsPregnant ewes at 121 days gestation underwent laparotomy as part of another research project.MethodsThermal and mechanical thresholds were measured before, and 2, 6, 24 and 48 hours after surgery. Thermal stimuli were delivered to the lateral aspect of the metatarsus via a skin-mounted probe, and mechanical stimuli to the contralateral site via a pneumatically driven 1.5 mm diameter pin. Each test was performed five times, alternating thermal and mechanical stimuli, with ten minutes between thermal stimuli. At the end of surgery ewes received either: 75 μg hour^?1 transdermal fentanyl patch (medial thigh) (group FP) (n = 8), or 3 μg kg^?1hour^?1 intra-peritoneal medetomidine via an osmotic pump (group IPM) (n = 8) inserted immediately prior to closure. Data were analysed using the Kruskal–Wallis RS Test (p < 0.05). Once a significant effect was identified, pairwise comparisons were performed using paired Wilcoxon RS tests. To compensate for multiple hypotheses testing, p < 0.005 was considered significant.ResultsPrior to surgery mean ± SD TT was 56.1 ± 5.0 °C (FP) and 55.6 ± 5.0 °C (IPM); MT was 5.3 ± 2.6 N (FP) and 8.0 ± 5.0 N (IPM). In FP there was no significant change in either TT or MT over time. In IPM there was no significant change in MT over time but TT increased at two hours to 59.2 ± 3.0 °C (p = 0.003). Skin temperature (ST) ranged from 33.0 to 34.7 °C and did not change over time. There were no significant differences between groups in TT, MT or ST.Conclusions and clinical relevanceAdministration of intra-peritoneal medetomidine (3 μg kg^?1hour^?1) by an osmotic pump increases the thermal nociceptive threshold in the immediate post operative period in pregnant sheep, suggesting that this agent may have a role in providing post-operative analgesia.     

Characterization of myofibrillar adenosine triphosphatase activity and liberation of actin from myofibrils upon heating chicken breast meat

Denaturation of actin and myosin in myofibrils induced by heating at 50°C was investigated to reveal the mechanism of irreversible liberation of actin from myofibrils on heating at lower temperatures than conventional cooking. Denaturation of these proteins was determined by Mg²⁺‐ATPase (adenosine triphosphatase) and Ca²⁺‐ATPase activities. When minced meat was heated for 20 min, actin was liberated accompanying denaturation of 80% of actin and 50% of myosin. Heating of the myofibrillar fraction (MFF) isolated from meat homogenate induced much slower denaturation of actin than myosin. When MFF was heated with sarcoplasmic fractions, denaturation of actin was facilitated, suggesting that sarcoplasmic fractions contain factors to facilitate actin denaturation. Inosine‐5′‐monophosphate, a component of sarcoplasmic fractions, was shown to have no effect on actin and myosin denaturation. These results suggest that heating meat at 50°C dissociates binding (‘Bond A’) between actin and myosin participating in ATPase activities, resulting in denaturation of both proteins under influence of sarcoplasmic components. Although denaturation of actin and myosin disrupted Bond A, actin was not liberated simultaneously, suggesting the presence of another bond (‘Bond B’, more heat‐stable than Bond A) between both proteins and necessity of disruption of Bond B for actin release from myofibrils.     

Physiological and haematological indices suggest superior heat tolerance of white-coloured West African Dwarf sheep in the hot humid tropics

Coat colour contributes to physiological adaptation in mammals and mediates response to thermal stress. Twenty-four adult West African Dwarf sheep of both sexes and with different coat colour types were used in this study. We measured rectal temperature (RT), respiratory rate (RR) and pulse rate (PR) before sunrise and sunset during the late dry season (January–March) and early rainy season (April–June) as well as packed cell volume (PCV), red blood cell (RBC) count, white blood cell (WBC) count, plasma sodium (Na⁺) and potassium (K⁺). Animals with black coat colour had the highest (P?<?0.05) mean values of 38.92?±?0.03 °C, 65.09?±?1.06 breaths/min, 81.35?±?0.78 beats/min, 1.70?±?0.01 for RT, RR, PR and heat stress index (HSI), respectively, followed by brown mouflon and brown with extensive white, while the Badger Face coloured sheep had the least mean values. There were significant (P?<?0.05) differences between male and female sheep for RT, RR, PR and HSI. Season had a significant (P?<?0.05) effect on RT, RR, PR and HSI. Coat colour and sex also significantly (P?<?0.01) affected RBC, WBC, Na⁺ and K⁺. Seasonal variation (P?<?0.05) in all the blood parameters was observed, with the exception of PCV. Interaction effect of coat colour and sex was significant (P?<?0.05) on RT and HSI. Correlation coefficients among the measured traits ranged from positive to negative values. These results indicate that selection of white-coloured sheep to attenuate heat stress is desirable in the hot humid tropics.     

The effects of L‐659,066, a peripheral α2‐adrenoceptor antagonist,and verapamil on the cardiovascular influences of dexmedetomidine in conscious sheep

Raekallio M. R., Honkavaara J. M., Vainio O. M. The effects of L‐659,066, a peripheral α₂‐adrenoceptor antagonist, and verapamil on the cardiovascular influences of dexmedetomidine in conscious sheep. J. vet. Pharmacol. Therap. 33 , 434–438. We investigated whether administration of L‐659,066, a peripheral α₂‐adrenoceptor antagonist, or verapamil, a calcium‐channel antagonist, would prevent the cardiovascular effects of dexmedetomidine. Eleven sheep received three intravenous treatments with a randomized, cross‐over design: dexmedetomidine (5 μg/kg, DEX); DEX with L‐659,066 (250 μg/kg, DEX + L); and verapamil (0.05 mg/kg) 10 min prior to DEX (Ver + DEX). Haemodynamics were recorded at intervals upto 40 min. Acute increases in mean arterial pressure (MAP) (106 ± 10.7 to 120.8 ± 11.7 mmHg), central venous pressure (CVP) (3.3 ± 3.2 to 14.7 ± 5.0 mmHg) and systemic vascular resistance (SVR) (1579 ± 338 to 2301 ± 523 dyne s/cm⁵), and decreases in cardiac output (CO) (5.36 ± 0.87 to 3.93 ± 1.30 L/min) and heart rate (HR) (88.6 ± 15.3 to 49.7 ± 5.5/min) were detected with DEX. The peak SVR remained lower after Ver + DEX (1835 ± 226 dyne s/cm⁵) than DEX alone, but the other parameters did not significantly differ between these treatments. 2 min after drug delivery, differences between DEX and DEX + L were statistically significant for all measured haemodynamic parameters. With DEX + L, an early decrease in MAP (99.9 ± 6.8 to 89.3 ± 6.6 mmHg) was detected, and DEX + L induced a slight but significant increase in CVP and a decrease in HR at the end of the observation period, while SVR and CO did not significantly change. All animals were assessed as deeply sedated from 2–20 min with no differences between treatments. L‐659,066 has great potential for clinical use to prevent the cardiovascular effects of dexmedetomidine mediated by peripheral α₂‐adrenoceptors, whereas the effects of verapamil were marginal.