Thermodynamic parameters of beta-lactoglobulin-pectin complexes assessed by isothermal titration calorimetry

Isothermal titration calorimetry (ITC) was used to determine the binding constant, stoichiometry, enthalpy, and entropy of beta-lactoglobulin/low- and high-methoxyl pectin (beta-lg-LM- and HM-pectin) complexes at 22 degrees C and at pH 4. The binding isotherms revealed the formation of soluble intrapolymer complexes (C1) further followed by their aggregation in interpolymer complexes (C2). The interaction between beta-lg and LM- or HM-pectin in C1 and C2 occurred spontaneously with a Gibbs free energy around -10 kcal/mol. The C1 were enthalpically driven, whereas enthalpic and entropic factors were involved in the C2 formation. Because ITC did not allow the dissociation of different enthalpic contributions, the values measured as pectin and beta-lg interacted could partially be attributed to conformational changes. The C1 had a binding stoichiometry of 8.3 and 6.1 beta-lg molecules complexed per LM- or HM-pectin molecule, respectively. The C2 had about 16.5 and 15.1 beta-lg molecules complexed per LM- and HM-pectin, respectively.     

Surgical results and fertility following correction of vesicovaginal reflux in mares

Objective To evaluate the surgical results and fertility of mares bred at various intervals relative to surgical management of urovagina. Design Sixty-one mares underwent surgery to correct vesicovaginal reflux. Surgery was performed at varying times relative to breeding and postoperative fertility was evaluated. Results Five mares were lost to follow-up and a further four were electively not bred. Of the remaining 52 mares, seasonal pregnancy and live foaling rates were 67% (35/52) and 54% (26/48), respectively. When mares were bred in the same cycle as the surgery, the first cycle following surgery, the second cycle following surgery or the following breeding season after surgery the seasonal pregnancy rates were 89% (8/9), 63% (10/16), 67% (2/3) and 63% (15/24), respectively. After censoring four mares that died of natural causes pre-foaling, the foaling rates were 88% (7/8), 50% (7/14), 0% (0/3), 52% (12/23), respectively. All mares bred in the same cycle as surgery or in the next cycle were bred once only that season, so the pregnancy rate per cycle of 72% (18/25) was identical to the seasonal pregnancy rate. Conclusions Good fertility (comparable to a normal population of mares) was achieved following surgery and the results suggest that delaying breeding until the following breeding season is not necessary. In addition, breeding in the same cycle as surgical repair is a previously unreported technique that should be considered to maintain a yearly foaling interval.     

Distal luxation of the patella in a horse

A 19-year-old Thoroughbred gelding presented with sudden onset, non-weight bearing lameness in the right hindlimb. Radiography confirmed distal luxation of the patella, which was replaced into its normal anatomical location under general anaesthesia. There were no pathological sequelae noted on follow-up examination 9 months after the initial injury. To our knowledge, this is a rare manifestation of patellar luxation, only reported once previously in the equine literature.     

Effect of density on brood size in noble crayfish,Astacus astacus L., subjected to indoor rearing conditions*

Abstract. The effects of density, cover, and individual versus communal holding on brood size and female survival during the breeding period were examined in a large-scale experiment, involving 3591 mature noble crayfish, Astacus astacus L., females. Crayfish density during both mating/spawning and winter incubation had major effects on egg survival. In the high density experiment (67 and 44 individuals per m² during mating/spawning and winter incubation respectively) mean pleopod egg number in April was 18-20. Lowering the density to 50 during mating/spawning (still 44 individuals per m² during winter incubation) gave mean pleopod egg numbers of 31-38. In the low density experiment (50 and 33 individuals per m² during mating/spawning and winter incubation) mean pleopod egg numbers were 44-46. Egg-bearing females held in individual compartments with perforated bottom had a mean pleopod egg number of 25. Female survival during winter incubation was high, ranging from 91.8 to 98.3%. Covering the basins had no distinct effect on egg and female survival. In Astacus astacus culture we recommend that crayfish density should not exceed about 50 individuals per m² during mating/spawning and about 33 individuals per m² during winter incubation.     

Effects of initial and extended exposure to an endophyte-infected tall fescue seed diet on faecal and urinary excretion of ergovaline and lysergic acid in mature geldings

Abstract

AIM: To determine the amount of ergovaline and lysergic acid retained or excreted by geldings fed endophyte-infected seed containing known concentrations of these alkaloids, and the effects of exposure time on clinical expression of toxicosis.

METHODS: Mature geldings (n=10) received diets containing either endophyte-free (E-) or endophyte-infected (E+) tall fescue seed during three experimental phases. The first phase (Days ?14 to ?1) was an adaptation phase, to allow all horses to adapt to a diet containing E- tall fescue seed. The second (Days 0 to 3) was the initial exposure phase to E+ tall fescue seed, used for the delivery of ergovaline and lysergic acid at 0.5 and 0.3 mg/kg of diet, respectively, to test the initial effects of exposure on routes and amounts of elimination of alkaloid. During this phase, half the geldings were exposed to an E+ diet while the rest served as controls by remaining on the E- diet. Once assigned to treatments, geldings remained on the same diet through the third phase (Days 4 to 21), which served as the extended exposure phase. Total outputs of faeces and urine were collected within each phase, to determine retention of ergovaline and lysergic acid and nutrient digestibility. Serum was collected weekly and analysed for activities of enzymes and concentrations of prolactin. Bodyweights (BW) and rectal temperatures were recorded weekly.

RESULTS: BW, rectal temperature, enzyme activities and concentrations of prolactin in serum, and nutrient digestibility were not affected by treatment. Total intake of ergovaline by geldings on the E+ diet was 3.5 and 3.6 (SE 0.20) mg/day, and 2.1 and 2.3 (SE 0.11) mg/day were not accounted for in initial and extended phases, respectively. Lysergic acid was excreted in the urine (4.0 and 4.9 (SE 0.97) mg/day) and faeces (2.5 and 2.7 (SE 0.35) mg/day) at greater amounts than that consumed (2.0 and 1.9 (SE 0.09) mg/day) during the initial and extended exposure phases, respectively. Animals exposed to E+ seed for a period of 20 days appeared to excrete more (1.5 vs 1.2 mg/day; SE 0.08; p=0.03) ergovaline in the faeces than those exposed for only 4 days.

CONCLUSIONS: Exposure time to the ergot alkaloids had a limited effect on the route of elimination or the amounts of ergovaline or lysergic acid excreted by horses. The primary alkaloid excreted was lysergic acid, and urine was the major route of elimination. These data will aid future research to improve animals′ tolerance to toxic endophyte-infected tall fescue.     

Copper deficiency in sheep: An assessment of the relationship between concentrations of copper in serum and plasma

AIM: To assess the relationship between concentrations of copper in serum and plasma in sheep.

METHODS: Concentrations of Cu were measured in paired serum and heparinised plasma samples collected from 110 sheep in nine flocks. Linear regression was used to evaluate whether flock or gender had a significant effect on the association between concentrations of Cu in serum and plasma. The individual results for concentrations of Cu in serum were then compared with those from plasma, using correlation and limits of agreement plotting.

RESULTS: Concentrations of Cu in serum ranged from 7.3 to 22 (mean 14.0) µmol/L, while concentrations in plasma ranged from 9 to 27 (mean 16.3) µmol/L. On average, concentrations of Cu in serum were 2.3 µmol/L lower than in plasma. Over the range of values seen in this study, concentrations of Cu in plasma and serum were significantly correlated (r=0.89), and mean concentrations in serum were 87% of those in plasma. There was no effect of flock or gender on the relationship between concentrations of Cu in serum and plasma. Despite the significant correlation, there was marked variability between individual samples in the proportion of Cu that was lost during clotting, with the 95% limits of agreement for serum Cu ranging from 70 to 104% of the plasma concentration.

CONCLUSIONS: As in cattle, the individual variability in the loss of Cu during clotting in sheep is too great for concentration of Cu in serum to be used as a substitute for that in plasma.

CLINICAL RELEVANCE: When assessing the blood Cu pool as part of the diagnosis of Cu-responsive disease in sheep, the concentration of Cu in plasma should be measured in preference to that of serum. We suggest that a range of 4.5 to 9 µmol/L in plasma be used to define marginal Cu status in sheep.