Derivation of clones close to met by preparative field inversion gel electrophoresis

The molecular analysis of genes identified by mutations is a major problem in mammalian genetics. As a step toward this goal, preparative field inversion gel electrophoresis (FIGE) was used to selectively isolate clones from the environment of genetically linked markers, and to select a subset of these clones containing sequences next to specific restriction sites rare in mammalian DNA. This approach has been used to generate a library highly enriched in sequences closely linked to the cystic fibrosis marker met. One clone derived from the end of a Not I restriction fragment containing the met sequence was analyzed in detail and localized within a long range map to a position 300 kilobase pairs 5' of the metD sequence.     

Epidemiological investigation of the buller steer syndrome (riding behaviour) in a western Canadian feedlot

Objective To describe the buller steer syndrome in a Western Canadian feedlot. Design A retrospective epidemiological study. Animals 78,445 male cattle that entered a 24,000-head feedlot in western Canada from 1991 to 1993. Procedure All cattle were given a hormonal growth promotant containing 20 mg oestradiol benzoate and 200 mg progesterone within 24 h of arrival at the feedlot. A ‘buller’ was a steer that was observed at daily pen checking to be ridden persistently by pen mates or had evidence of having been persistently ridden by pen mates. At the completion of the feeding period, animal health records for bullers were collected and analysed. Results The prevalence of bullers in the total population was 2139/78,445 (2.7%, range per pen 0 to 11.2%). The prevalence of bullers increased with increasing weight and age. The relapse risk after first treatment (three days in the feedlot hospital plus treatment for concurrent disease) was 30% on average (27 to 35%). Individual records from 9734 yearling steers that entered the feedlot in 1991 and 1992 showed that bullers were significantly (P < 0.05) heavier at processing than non-bullers. Bullers occurred as a point source epidemic with the cause occurring soon after cattle arrived at the feedlot and were mingled into pen groups. This gave a ‘days on feed’ distribution. The peak incidence of bullers occurred much sooner after arrival and dropped off much quicker in older cattle. The daily incidence of bullers was temporal, but was not related to season of the year, weather conditions or any other feedlot management practice. It was related to the seasonal arrival of cattle at the feedlot, their age at entry to the feedlot and the post arrival occurrence of bullers. Reimplantation with hormonal growth promotants and castration of intact bulls did not produce an epidemic of bullers. Conclusion The findings of this study support the theory that bullers are the result of agonistic interactions, which occur concurrent with the establishment and maintenance of a social hierarchy within pens of feedlot cattle.     

Precambrian sponges with cellular structures

Sponge remains have been identified in the Early Vendian Doushantuo phosphate deposit in central Guizhou (South China), which has an age of approximately 580 million years ago. Their skeletons consist of siliceous, monaxonal spicules. All are referred to as the Porifera, class Demospongiae. Preserved soft tissues include the epidermis, porocytes, amoebocytes, sclerocytes, and spongocoel. Among thousands of metazoan embryos is a parenchymella-type of sponge larvae having a shoe-shaped morphology and dense peripheral flagella. The presence of possible amphiblastula larva suggests that the calcareous sponges may have an extended history in the Late Precambrian. The fauna indicates that animals lived 40 to 50 million years before the Cambrian Explosion.     

Variation in the immune status of two Australian pig breeds

Objective To investigate the variation in immune competence of two Australian pig breeds.
Design A panel of immune tests were used to assess breed and sire differences in weaner piglets of Large White and Duroc breeds.
Procedure All piglets were immunised against porcine leptospirosis. Blood samples were taken for studies on lymphocyte phenotypes, mitogenic responses of blood cells and serological analysis.
Results Significantly larger blood leucocyte numbers were found in Large White piglets compared with Duroc piglets after vaccinations. No significant difference in concanavalin A induced blood cell proliferation was found between these two breeds before or after vaccinations. Some significant breed variation in blood lymphocyte phenotypes was found. While the age-related changes of lymphocyte phenotypes were similar for the two breeds, the Large White breed had significantly larger numbers of CD2+ and CD4+ cells than the Duroc breed after the two vaccinations. There were also significant sire effects on CD8+ cells within the Large White breed after the first vaccination. No significant breed difference was detectable in serum IgG concentrations but sire differences within each breed before the primary vaccination were found. The serum antibody response to vaccination against leptospirosis was generally small, and showed no variations due to either breed or sire. No gender effects were found during the entire study.
Conclusion The study demonstrated significant differences in some important immune components of the pig breeds studied. This may in turn indicate the variation in their immune competence or disease resistance. However, further investigation into the heritability and correlation with specific immune responses is required.     

Musculoskeletal responses of 2-year-old Thoroughbred horses to early training. 4. Morphometric,microscopic and biomechanical properties of the digital tendons of the forelimb

AIM: To determine the weight, volume, density and cross-sectional area (CSA) of the digital flexor tendons, common digital extensor tendon (CDET) and suspensory ligament (SL) of the forelimb of young Thoroughbred horses in early training, and to assess the response to a training programme of known parameters of superficial digital flexor tendon (SDFT) tissue at mid-metacarpal level.

METHODS: The tendons of seven 2-year-old Thoroughbred horses in training were inspected, transected into segments of known length, and compared with those from seven untrained horses matched for age, sex and breed. The weight, volume, density and CSA of each segment, and the crimp angle, histological features, and biomechanical ultimate stress and stiffness of tendon samples from the mid-metacarpal region of the SDFT were determined.

RESULTS: There was no macroscopic evidence of swelling or discolouration in any of the tendon segments or cut surfaces. The volumes of SDFT and CDET segments of horses in the trained group were significantly greater than those in the untrained group (p=0.036 and p=0.039, respectively). A greater increase in volume than weight resulted in a lower density in the SDFT but not CDET in trained compared with untrained horses (p=0.038). CSA of these two tendons was significantly greater in the trained group (p=0.002 and 0.036, respectively), the percentage increase being greater in the CDET than the SDFT. The number of tenocytes at four sites in the mid-metacarpal SDFT region was less in trained than untrained horses (p=0.025). There was no histological evidence of inflammation, and no difference in crimp angle between groups. There was no significant between-group difference in stiffness or ultimate stress of tendon strips.

CONCLUSIONS: Volume and CSA of the SDFT and CDET were larger in trained than untrained horses. The SDFT was less dense in the trained group. Because no evidence of tendonitis was detected and training appeared to have no significant effect on crimp angle or biomechanical properties of tendon strips, the size and density changes were presumed to be adaptive and induced by the training.

CLINICAL RELEVANCE: Although evident in this in vitro study, the detection of adaptive from initial pathological increase in size of the SDFT is likely to be difficult in vivo.     

The effect on creatinine kinase activity of freezing plasma from healthy foals

Abstract

AIM: To assess the stability of creatine kinase (CK) activity in plasma collected from healthy foals and frozen at ?20°C for up to 12 weeks.

METHODS: Samples of venous blood drawn from 25 foals were analysed for CK activity soon after collection, and again after 1 and 12 weeks of freezing at ?20°C.

RESULTS: CK activity decreased (p<0.001) between Week 0 and Week 1 and between Week 0 and Week 12.

CONCLUSIONS AND CLINICAL RELEVANCE: Decreases in CK activity were statistically significant but clinically insignificant.     

Azospirillum — plant root associations: A review

Summary Bacteria of the genus Azospirillum are extensively studied for their plant-growth promoting effect following inoculation. Physiological and biochemical studies of these diazotrophic bacteria are now benefiting from recent breakthroughs in the development of genetic tools for Azospirilum. Moreover, the identification and cloning of Azospirillum genes involved in N₂ fixation, plant interaction, and phytohormone production have given new life to many research projects on Azospirillum. The finding that Azospirillum genes can complement specific mutations in other intensively studied rhizosphere bacteria like Rhizobia will certainly trigger the exploration of new areas in rhizosphere biology. Therefore a review of the Azospirillum-plant interactions is particularly timely.     

Antioxidant enzyme activities,plasma hormone levels and serum metabolites of finishing broiler chickens reared under high ambient temperature and fed lemon and orange peel extracts and Curcuma xanthorrhiza essential oil

The negative effects of high ambient temperature during some months of the year on poultry production have been of great concern in many countries. Dietary modifications are among the most practical ways to alleviate the effects of high temperature. Possible effects of dietary supplementation with 200 or 400 mg/kg feed of lemon peel extract (LPE), orange peel extract (OPE) and Curcuma xanthorrhiza essential oil (CXEO) under hot conditions (34 °C with 50% relative humidity for 5 h daily starting from day 28 until day 38 of age) on blood antioxidant enzyme activities, biochemical parameters and antibody titres of broiler chickens were investigated. All extracts are rich in phenolic compounds and highly available. Compared to control, supplementation with OPE at 400 mg/kg and CXEO significantly increased erythrocyte glutathione peroxidase and superoxide dismutase activity, plasma growth hormone concentrations and serum phosphorus, total protein and chloride concentrations and decreased serum low‐density lipoprotein and cholesterol concentrations in chickens at 38 days of age. Regarding antibody titres, CXEO supplementation at 400 mg/kg caused a significant increase in bronchitis antibody titres. Supplementation with LPE and OPE gave more inconsistent results. Most interesting, 400 mg/kg LPE significantly increased 3,5,3′‐triiodothyronine and GH concentration as compared to the control. In conclusion, the herbal extracts tested in this study, in particular CXEO at 400 mg/kg, may relieve some of the changes in blood composition induced by increased ambient temperatures.