Insulin-like growth factor I receptor analysis of satellite cell-derived myotube membranes established from two lines of Targhee rams selected for growth rate

Ovine-derived fibroblasts were used to validate an insulin-like growth factor I (IGF-I) membrane-receptor binding assay system. Competitive binding using fibroblasts revealed that half-maximal inhibition of 125I-IGF-I binding by IGF-I was 2.3 nM. SDS-polyacrylamide gel electrophoresis analysis of specific protein-associated 125I-IGF-I was consistent with the migration of 125I-IGF-I-labeled Type I IGF receptor alpha-subunits at Mr 133,000 daltons. Further, the efficiency of two cell solubilization methods was examined and time-dependent binding equilibrium was determined for the membrane assay system. Satellite cell-derived myotubes were subsequently isolated from primary satellite cell cultures established from the semimembranosus muscles of high and low efficiency-of-gain (EOG) Targhee rams, and IGF-I receptor dynamics were measured. A membrane competitive binding study revealed that half-maximal inhibition of 125I-IGF-I binding was achieved by 1-ng IGF-I for low, and 10-ng IGF-I for high, EOG myotube membrane preparations. Kd values were similar between the high EOG (4.78 nM) and low EOG (2.95 nM) groups; however, receptor concentrations (Bmax) appeared to differ between groups. High EOG membrane receptor Bmax was 3.88 pmole/micrograms protein (19.87 pmole/micrograms DNA), whereas low EOG membrane receptor Bmax was 1.22 pmole/micrograms protein (9.28 pmole/micrograms DNA). These preliminary findings support the hypothesis that genetic selection for EOG results in altered satellite cell responsiveness to IGF-I.     

Pharmacokinetics of articaine hydrochloride and its metabolite articainic acid after subcutaneous administration in red deer (Cervus elaphus)

AIM: To develop and validate a simple and sensitive method using liquid chromatography-mass spectrometry (LC-MS) for quantification of articaine, and its major metabolite articainic acid, in plasma of red deer (Cervus elaphus), and to investigate the pharmacokinetics of articaine hydrochloride and articainic acid in red deer following S/C administration of articaine hydrochloride as a complete ring block around the antler pedicle.

METHODS: The LC-MS method was validated by determining linearity, sensitivity, recovery, carry-over and repeatability. Articaine hydrochloride (40?mg/mL) was administered S/C to six healthy male red deer, at a dose of 1?mL/cm of pedicle circumference, as a complete ring block around the base of each antler. Blood samples were collected at various times over the following 12 hours. Concentrations in plasma of articaine and articainic acid were quantified using the validated LC-MS method. Pharmacokinetic parameters of articaine and articainic acid were estimated using non-compartmental analysis.

RESULTS: Calibration curves were linear for both articaine and articainic acid. The limits of quantifications for articaine and articainic acid were 5 and 10?ng/mL, respectively. Extraction recoveries were >72% for articaine and >68% for articainic acid. After S/C administration as a ring block around the base of each antler, mean maximum concentrations in plasma (C_max) of articaine were 1,013.9 (SD 510.1) ng/mL, detected at 0.17 (SD 0.00) hours, and the C_max for articainic acid was 762.6 (SD 95.4) ng/mL at 0.50 (SD 0.00) hours. The elimination half-lives of articaine hydrochloride and articainic acid were 1.12 (SD 0.17) and 0.90 (SD 0.07) hours, respectively.

CONCLUSIONS AND CLINICAL RELEVANCE: The LC-MS method used for the quantification of articaine and its metabolite articainic acid in the plasma of red deer was simple, accurate and sensitive. Articaine hydrochloride was rapidly absorbed, hydrolysed to its inactive metabolite articainic acid, and eliminated following S/C administration as a ring block in red deer. These favourable pharmacokinetic properties suggest that articaine hydrochloride should be tested for efficacy as a local anaesthetic in red deer for removal of velvet antlers. Further studies to evaluate the safety and residues of articaine hydrochloride and articainic acid are required before articaine can be recommended for use as a local anaesthetic for this purpose.     

Policies for the vaccination of cats and dogs in New Zealand veterinary practices

AIMS: To determine current practices and attitudes towards vaccination of dogs and cats of veterinarians in New Zealand; the methods used for informing clients on which vaccines to use, and the preferred site for vaccination of cats.

METHODS: A postal questionnaire was sent to all 483 listed veterinary practices in New Zealand during February 2012. Some questions were specific to pet dogs, cats, or working farm dogs. Responses were categorised according to practice type and geographical region of the respondent. Factors associated with respondent recommendation of annual vaccination with modified live viral (MLV) vaccines were examined using logistic regression analysis. Vaccines that were considered to be essential for every animal were defined as core; those that may be recommended for animals whose location or lifestyle placed them at risk, were defined as non-core.

RESULTS: There were 204 useable returns, equivalent to a response rate of 42.2%, distributed across the country. Annual vaccination with MLV vaccines of dogs was recommended by 54/198 (27.3%) respondents, and of cats by 107/181 (59.1%) respondents. Factors associated with the recommendation of annual administration of MLV vaccines to dogs included being a companion animal practice, a desire for policies on vaccination to be left to individual clinics, and having one veterinarian in the practice. Administration of the final vaccination for puppies was recommended at ≥14 weeks old by 55/185 (29.7%) respondents, and for kittens at ≥13 weeks old by 42/183 (23%) respondents. Of respondents that administered MLV vaccines annually, 62/103 (60.2%) believed reducing the frequency of vaccination would reduce income, and 52/103 (50.5%) considered it would have a negative effect on animal health. Advice to enable clients to decide which non-core vaccines were administered was given by 181/199 (91%) respondents. Factors considered when recommending a vaccine included consideration of risk to individual patients (190/203; 93.6%), requirements of boarding kennels/catteries (165/203; 81.3%) and clinic vaccination policy (142/203; 70%). The preferred site for administering MLV vaccines to cats was the dorsal neck or inter-scapular region (137/198; 69.2%). Amongst respondents, 18 wanted disease surveillance information to allow for truly informed decisions to be made about vaccination.

CONCLUSIONS AND CLINICAL RELEVANCE: Veterinarians can now compare their own vaccination practices and attitudes with those of veterinarians nationally, and internationally. There is a need for national surveillance information and for continued education of the public and commercial kennel and cattery owners for optimal vaccination strategies to be developed.     

Attitudes and perceptions of veterinary paraprofessionals in New Zealand to postoperative pain in dogs and cats

AIM: To survey the attitudes and perceptions of veterinary paraprofessionals in New Zealand to postoperative pain in dogs and cats.

METHODS: In December 2011, veterinary paraprofessionals (VP) from throughout New Zealand were invited to participate in an online survey. Eleven questions, which were divided into five sections, were used to determine demographic information, the respondents’ assessment of pain after commonly performed surgeries in dogs and cats, their opinions on provision of analgesia, who had responsibility for pain monitoring and the use of any formal pain scoring system in the practice.

RESULTS: Data from 165 respondents were able to be used, and 162 (98%) respondents to the survey were female. According to the respondents’ estimates, fracture repair in dogs and repair of diaphragmatic hernias in cats had the highest pain score following surgery. Neutering procedures involving dogs were scored higher than for cats (p<0.01). All respondents agreed that animals benefit from perioperative analgesia. The veterinary nurse was reported to be predominantly responsible for monitoring pain in animals postoperatively by 116/165 (70.3%) respondents. Of 165 respondents, 154 (93%) considered that their knowledge of pain and assessment of pain could be enhanced.

CONCLUSIONS: This survey reflects the attitudes and perceptions of a sample of VP in New Zealand to postoperative pain in dogs and cats. The results indicate that all respondents believe that surgery results in sufficient pain to warrant analgesic therapy. Routine neutering surgeries were considered to be more painful in dogs than in cats. The current survey also provides information to educators on potential areas of focus, given that 93% of respondents felt that their knowledge of pain and assessment of pain could be enhanced.     

Effects of thyroxine and growth hormone treatment of dairy cows on milk yield, cardiac output and mammary blood flow

Four cows received thyroxine injections (T4; 20 mg/d) and three cows received growth hormone injections (GH; 44 mg/d) for 4 d during successive 16-d experimental periods. Measurement was made of milk yield, protein yield, mammary tyrosine and phenylalanine uptake, blood plasma hormone concentrations, mammary blood flow and cardiac output. Milk yield increased by 25% with T4 and 21% with GH treatment. Milk protein content tended to decline during T4 treatment and increase following GH treatment. Cardiac output increased by 8.9 liter/min (20%) and 4.6 liter/min (10%) with T4 and GH injection. Mammary blood flow (half-udder) increased from 3.6 to 4.9 liter/min (35%) and from 3.3 to 4.4 liter/min (33%) with T4 and GH treatment, respectively. These increases calculated on a whole-udder basis, accounted for 28% (T4) and 48% (GH) of the increases in cardiac output. The proportion of cardiac output perfusing the (whole) udder increased to 19.1% (T4) and 18.7% (GH), increases of 17 and 30%, respectively. Heart rate increased with T4 (but not GH treatment) from 80 to 115/min. Ratio of blood flow to milk yield was not changed by either treatment. The proportion of cardiac output perfusing the udder likely plays a major role in facilitating the partitioning of nutrients for milk synthesis.     

Protein and fat utilization in lactating sows: I. Effects on milk production and body composition

In order to provide data with which to challenge a model of metabolism of lactating sows, we conducted a study to determine milk production and body and mammary composition in sows consuming a range of energy and amino acid intakes and nursing 11 to 12 pigs. Sows (2nd through 4th parity) consumed the same ration during gestation and consumed 6.1 kg/d (as-fed) for a 20 d lactation. Litter size was standardized at 12 pigs within 3 d of farrowing. Diets were formulated to provide three different amounts of protein intake and two different amounts of fat intake. Protein intakes of sows in high (HP), medium (MP), and low protein (LP) treatment groups were 863, 767, and 678 g/d with 59, 53, and 47 g/d lysine at two levels of fat intake, 117 (LF) and 410 g/d (HF). Number of pigs weaned per litter was 11.4 +/- 0.5 and milk production and litter weight gain was less (P < 0.01) in the last week of lactation for sows consuming the least protein. Medium and low protein intakes increased (P < 0.05) loss of body lean and protein. Change in carcass protein during lactation was -1.4, -3.0, -2.2, -1.2, -1.9 and -2.1 kg (SD 2.6) for sows fed HPLF, MPLF, LPLF, HPHF, MPHF, and LPHF. Body fat (carcass and visceral) change was 0.4, -3.7, -4.1, -0.3, 3.4, and -1.3 kg (SD 6.6) in HPLF, MPLF, LPLF, HPHF, MPHF, and LPHF groups. Total amount of mammary parenchyma increased more (P < 0.05) in sows fed a higher fat diet. These data are consistent with general knowledge of changes in body composition in lactation of sows. However, changes in body protein and fat were correlated across treatments and different from that reported for sows nursing smaller litters. These data help our quantitative understanding of nutrient flux in sows nursing large litters and allow a severe challenge of existing models of metabolism in sows.     

Pharmacokinetics of penicillin G procaine versus penicillin G potassium and procaine hydrochloride in horses

OBJECTIVE: To compare the pharmacokinetics of penicillin G and procaine in racehorses following i.m. administration of penicillin G procaine (PGP) with pharmacokinetics following i.m. administration of penicillin G potassium and procaine hydrochloride (PH). ANIMALS: 6 healthy adult mares. PROCEDURE: Horses were treated with PGP (22,000 units of penicillin G/kg of body weight, i.m.) and with penicillin G potassium (22,000 U/kg, i.m.) and PH (1.55 mg/kg, i.m.). A minimum of 3 weeks was allowed to elapse between drug treatments. Plasma and urine penicillin G and procaine concentrations were measured by use of high-pressure liquid chromatography. RESULTS: Median elimination phase half-lives of penicillin G were 24.7 and 12.9 hours, respectively, after administration of PGP and penicillin G potassium. Plasma penicillin G concentration 24 hours after administration of penicillin G potassium and PH was not significantly different from concentration 24 hours after administration of PGP. Median elimination phase half-life of procaine following administration of PGP (15.6 hours) was significantly longer than value obtained after administration of penicillin G potassium and PH (1 hour). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that i.m. administration of penicillin G potassium will result in plasma penicillin G concentrations for 24 hours after drug administration comparable to those obtained with administration of PGP Clearance of procaine from plasma following administration of penicillin G potassium and PH was rapid, compared with clearance following administration of PGP.     

Follicular Dynamics in Heifers during Pre-pubertal and Pubertal Period Kept under Two Levels of Dietary Energy Intake

The objective of this study was to characterize follicular dynamics in pre-pubertal, pubertal and post-pubertal periods, as well as the effect of high-energy intake on follicular development and age at puberty in heifers. Thirty-one Nelore (Bos indicus) heifers, 6 months old, were randomly assigned to receive two different diets: one of low (GI) and other of high dietary energy intake (GII). Animals were evaluated in relation to body weight gain by being weighed every 21 days. Heifers were evaluated every other day by real-time linear ultrasonography to characterize ovarian structures development from weaning to post-pubertal period. Blood samples were collected to determine plasmatic concentrations of progesterone by RIA method. The ovulation was determined when progesterone concentrations were >1 ng/mL in three consecutive samples, and by ultrasound images of corpus luteum; and oestrous behaviour in some animals. Age at puberty differed among heifers of GII (17.00 +/- 0.46 months) compared with heifers of GI (19.87 +/- 0.47 months; p < or = 0.05). Maximum size of the dominant follicles at pre-pubertal period was greater in GII heifers than in GI (10.52 +/- 0.33 and 9.76 +/- 0.15 mm, respectively; p < or = 0.05). As heifers approached first ovulation time, size of dominant follicle increased (11.75 +/- 0.37 mm for GI and 12.52 +/- 0.91 mm for GII; p < or = 0.05). Body weight at puberty was not different in both groups (302.33 +/- 27.31 kg for GI and 326.19 +/- 27.78 kg for GII heifers; p > 0.05). We conclude that animals receiving high dietary energy intake attained the puberty earlier and the development of follicles were different than in low dietary energy intake.