Synthetic polypeptide antagonists of the hypothalamic luteinizing hormone releasing factor

Two analogs of the hypothalamic luteinizing hormone releasing factor modified at the histidine-2 position were tested for biological activity (secretion of luteinizing hormone) in cultures of dispersed rat anterior pituitary cells. The analog in which glycine was substituted for histidine at position 2, [Gly(2)]LRF, behaves as a partial agonist releasing less than 50 percent of the luteinizing hormone secreted at maximum concentrations of the releasing factor, while the analog in which histidine at position 2 is deleted has no significant agonist activity at any of the doses tested. When added to the cultured cells at molar ratios 10(3) to 10(4) times that of the luteinizing hormone releasing factor, both analogs decrease the amount of luteinizing hormone secreted in response to the releasing factor.     

Vitamin E and polyunsaturated fatty acids in bovine muscle and the oxidative stability of beef from cattle receiving grass or concentrate-based rations

The present study was designed to assess the balance between antioxidant and prooxidant components and the oxidative stability of beef from cattle fed exclusively grazed pasture (PAS) or a barley-based concentrate offered indoors (CONC) for 11 mo, or fed grass silage indoors for a 5-mo winter period, followed for the remaining 6-mo summer period by grazed pasture (SiP) or by grazed pasture plus concentrate at 50% of the dietary DM (SiPC). Muscle prooxidant and antioxidant components were determined by measuring fatty acids and α-tocopherol concentration of LM, respectively. Lipid oxidation and color stability were monitored in ground LM, packaged in a high-oxygen modified atmosphere, over 11 d of refrigerated storage. Vitamin E concentration decreased (P < 0.0005) with an increasing proportion of concentrate in the diet (2.59, 2.45, 1.76, and 1.15 μg/g for PAS, SiP, SiPC, and CONC, respectively). A greater proportion of PUFA was found in LM from cattle in the PAS, SiP, and SiPC groups compared with animals in the CONC group (9.62, 11.04, 8.96, and 6.94%, respectively; P < 0.0005). A greater concentration of highly peroxidizable PUFA was found in LM from heifers in the PAS, SiP, and SiPC groups compared with those in the CONC group (0.84, 0.85, 0.87, and 0.65 mg/g of muscle, respectively; P = 0.02). Dietary treatment affected lipid oxidation (P < 0.0005), with greater 2-thiobarbituric acid reactive substance values in beef from heifers in the SiPC group than in beef from those in the PAS, SiP, and CONC groups. Dietary treatment affected myoglobin oxidation (P = 0.002) during storage, with greater metmyoglobin accumulation in beef from animals receiving concentrate (CONC and SiPC treatments) than in beef from cattle in the PAS and SiP groups. Consequently, feeding concentrate impaired meat color stability over the storage duration, with greater H* (hue angle) values (P < 0.0005) in meat from heifers in the SiPC and CONC groups compared with meat from those in the PAS and SiP groups. The results of the present study confirm a positive effect of grass-based feeding systems on meat color stability compared with concentrate-based dietary strategies. It appears that vitamin E in muscle alone does not explain the resistance of meat to oxidative deterioration because a clear interaction with highly peroxidizable PUFA exists.     

Isolation and molecular characterisation of Neospora caninum in cattle in New Zealand

AIM: To isolate Neospora caninum from the brains of naturally infected cattle and use molecular techniques to characterise the isolates.

METHODS: Neospora caninum tachyzoites were isolated in Vero cell culture from the brains of a cow and two calves. The isolates were characterised using polymerase chain reaction (PCR) methods, DNA sequencing, an immunofluorescent anti-body test (IFAT), transmission electron microscopy (TEM), and immunohistochemistry (IHC). The brains of the three cattle were subjected to histopathological examination. A pathogenicity study was conducted in 120 BALB/c mice.

RESULTS: Neospora caninum tachyzoites were isolated from all three cases and first observed in vitro between 14 and 17 days post-inoculation. Parasites were sub-cultured and maintained in Vero cell culture for more than 6 months. PCR products were generated for all three isolates, using two different primers. Sequencing of the PCR products and a subsequent BLAST search identified the isolates as N. caninum. In addition, the isolates tested positive using IFAT and IHC, and ultrastructure revealed by TEM was characteristic of N. caninum. Histopathological examination revealed lesions characteristic of N. caninum in 1/3 brains. In the pathogenicity study using BALB/c mice, the mortality rate was 3–7%.

CONCLUSION: This was the first successful isolation of N. caninum in New Zealand confirmed using molecular characterisation tests.     

Production benefits from pre- and post-lambing anthelmintic treatment of ewes on commercial farms in the southern North Island of New Zealand

Abstract

AIMS: To measure the magnitude and variability in production responses to anthelmintic treatments administered to adult ewes around lambing.

METHODS: Ewes carrying twin lambs, from sheep and beef farms (eight in Year 1 and six in Year 2) in the Wairarapa region of New Zealand, were enrolled in 14 trials (part of an experiment carried out on one farm in one year). Experiment 1 compared ewes treated 2–4?weeks pre-lambing with a controlled release capsule (CRC) containing abamectin, albendazole, Se and Co, to ewes injected pre-lambing with a long-acting Se plus vitamin B₁₂ product, and to untreated ewes. Experiment 2 included these treatments, plus a CRC administered at pregnancy scanning. Experiment 3 included the same treatments as Experiment 1, plus administration of a CRC containing albendazole, Se and Co, injectable moxidectin or oral derquantel plus abamectin, all administered pre-lambing, or oral derquantel plus abamectin administered 4–6?weeks after lambing. Variables compared were ewe liveweight at weaning and pre-mating, lamb liveweight at weaning, total weight of lamb weaned per ewe and ewe dag score at weaning.

RESULTS: Ewes treated with a CRC pre-lambing were heavier than untreated ewes (mean 3.2?kg) at weaning in 12/14 trials, and pre-mating (mean 2.8?kg) in 9/14 trials (p<0.001). Compared with mineral-treated ewes the mean difference was 2.8?kg pre-lambing (9/14 trials) and 1.7?kg pre-weaning (6/14 trials). Lambs reared by treated ewes were heavier (mean 1.55?kg) at weaning in 6/14 trials (p<0.001), but there was no effect of CRC treatment on total weight of lambs weaned per ewe (p=0.507). Variation in weight of lamb weaned per ewe was largely explained by differences in lamb survival from birth to weaning (p<0.001), with no effect of CRC treatment (p>0.65).

Treatment of ewes with a CRC at pregnancy scanning was neither better nor worse than a pre-lambing treatment (p=0.065).

There was no difference in the response from treatment with either of the two CRC or moxidectin. Treatment with short-acting oral anthelmintics resulted in no consistent benefit.

CONCLUSIONS: Anthelmintic treatments administered to ewes around lambing resulted in variable responses between farms and years, which in some trials were negative for some variables, and some of the variability was due to the mineral component of the CRC. The widespread perception amongst farmers and veterinarians that anthelmintic treatment of ewes around lambing will always result in positive benefits is not supported.     

An ethogram of post-anesthetic recovery behaviors in horses: comparison of pre- and post-anesthetic behaviors

Pain management is an important post-operative concern. Pain scales may rely on the observer's subjective assessment of the level of discomfort and may not correlate with physiologic or pharmacologic measures of pain. The purpose of this study was to develop an objective measure of behavior in healthy pain-free horses recovering from anesthesia that could be used for comparison with the behavior of horses recovering from a surgical procedure.
Focal sampling with videotape and observation was done on five healthy horses before anesthesia to establish baselines. Behavioral measures included head turns, tail swishes, eyelid aperture/size, ear position, angle of neck, weight shifts, and ambulation. Physiologic measures included heart rate, respiratory rate, and temperature. The horses were anesthetized for 2 hours with isoflurane, and in the recovery stall, data were collected continuously on videotape from the time of extubation to standing. Focal sampling of 15 minutes was repeated at 1, 2, 4, and 24 hours after the horses returned to their stalls. Video data were analyzed using the Noldus Observer Video Analysis System.
A wide variation in behavior was observed between horses in the recovery stall. Observations at 1, 2, and 4 hours revealed change from the up/forward ear position to the down ear position, and a decrease in weight bearing of the hind limb from a baseline of 11–12 minutes out of 15 minutes to 7–9 minutes with an increase in toe pointing. Time spent standing was similar to baseline of 93.8%, and the neck angle was not changed from baseline of 173.5°. Values had returned to baseline at 24 hours.
Changes in behavior were induced by anesthesia alone and must be taken into consideration when evaluating analgesic treatments.     

Cholelithiasis and hyperthyroidism in a cat

A 14-year-old domestic short-hair cat presented with a history of intermittent malaise and increased drinking. A diagnosis of hyperthyroidism and cholelithiasis was made by a combination of blood testing, radiography and ultrasonography. After medical management of hyperthyroidism, thyroidectomy and cholecystectomy were successfully performed. Removed choleliths were comprised of calcium carbonate and bilirubinate. Histopathological analysis of tissue suggested low grade pancreatic and hepatobiliary disease, as well as hyperthyroidism, might have contributed to stone formation.     

Acceptance by stoats (Mustela erminea) of 1080 (sodium monofluoroacetate) in small-volume baits and its effect on behaviour and time to death

AIMS: To assess whether stoats (Mustela erminea) would eat small baits containing 0.1% sodium monofluoroacetate (1080); whether they would die from it; how long it would take to kill them; and to document the behaviour of 1080-intoxicated stoats.

METHODS: Stoats were offered 1-g baits of two semi-fluid formulations containing 0.1% 1080, presented in open dishes, and their subsequent behaviour was monitored by video and direct observation. Muscle samples from stoats that died were analysed for 1080 residues.

RESULTS: There was no significant difference between two types of bait with regard to acceptance, mortality, and time to death, and behavioural effects were similar; consequently, results from the two types of bait were combined. Twelve of 14 stoats offered the baits ate them voluntarily, and a 13th licked bait off its fur; all 13 died between 1 h 15 min and 4 h 7 min (mean 2 h 38 min) later. At first (range 29 min — 2 h 7 min, mean 1 h 1 min), their behaviour appeared to be normal. Ataxia and hyperactivity were the first behavioural signs of poisoning, and lasted 2 min — 1 h 40 min (mean 26 min). This was followed by recumbency with convulsions and rapid breathing (range 16 min to 2 h, mean 58 min), then recumbency with limited activity and progressively shallow breathing prior to death (range 1–51 min, mean 33 min). Stoats became non-responsive to a light being turned on, or to touch once recumbency became sustained.

Residues of 1080 were found in muscle tissue of all 13 dead stoats, at concentrations ranging from 0.075 μg/g in a 287-g male that died 4 h 7 min after eating only 0.74 g of bait, to 2.5 μg/g in a 254-g female that died 2 h 42 min after taking a whole 1-g bait.

CONCLUSION: Stoats will voluntarily take small (1-g) baits containing a lethal dose of 1080 at 0.1%, and die from it comparatively rapidly for a mammalian carnivore.