Intracerebroventricular Infusion of Leptin into Mature Merino Rams of Different Metabolic Status: Effects on Blood Concentrations of Glucose and Reproductive and Metabolic Hormones

In mature Merino rams, nutrition is one of the external cues that most strongly affects the reproductive centres of the preoptic-hypothalamic continuum. The signalling pathways that link dietary status and the activity of the neurones that produce gonadotrophin-releasing hormone signals are thought to be partly hormonal in nature to reflect the amount of body reserves. Among the hormones thought to be involved are insulin and leptin. This study tested whether recombinant bovine leptin infused (0.4 microg/h) into the third cerebral ventricle would stimulate pulsatile luteinizing hormone (LH) secretion in mature Merino rams when their energy status was low or decreasing, during both chronic (fasting) and acute reductions of energy balance. Leptin may interact with other hormones that depend on energy availability, so we also monitored changes in circulating concentrations of insulin, thyroid hormones, growth hormone, prolactin and adrenocorticotrophin. Overall, our data do not support this hypothesis. The dietary regimes induced clear responses in the metabolic profiles of the animals but there was no clear effect of central leptin administration on LH pulse frequency. The relationships between the hormonal systems measured in the present study add weight to the contention that leptin plays only a permissive role in the nutritional control of the reproductive axis and that other hormonal signals (particularly insulin) or pathways are acting in concert with leptin to stimulate the reproductive axis.     

Mycobacterium salmoniphilum infection in farmed Atlantic salmon, Salmo salar L

Multiple greyish‐white visceral nodules containing abundant rapidly growing and acid‐fast bacteria, subsequently identified as Mycobacterium salmoniphilum, were detected in moribund and newly dead market‐sized fish during a period of increased mortality in an Atlantic salmon, Salmo salar, farm in western Norway. Isolates cultured from diseased fish were phenotypically consistent with Mycobacterium sp. previously isolated from Atlantic salmon [MT 1890 (= NCIMB13533), MT1892, MT1900 and MT1901] in the Shetland Isles, Scotland. Partial sequences of 16S rDNA, ribosomal RNA internal transcribed spacer (ITS1), 65‐kDa heat‐shock protein (Hsp65) and β subunit of RNA polymerase (rpoB) revealed 97‐99% similarity with M. salmoniphilum type strain ATCC 13758^T. The source of infection was not confirmed. Koch’s postulates were fulfilled following experimental challenge of Atlantic salmon with field isolate NVI6598 ( FJ616988 ). Mortality was recorded in experimentally infected fish; however, the infection remained subclinical in the majority of affected fish over the 131‐day challenge period.     

Protein displacement by DExH/D "RNA helicases" without duplex unwinding

Members of the DExH/D superfamily of nucleic acid-activated nucleotide triphosphatases are essential for virtually all aspects of RNA metabolism, including pre-messenger RNA splicing, RNA interference, translation, and nucleocytoplasmic trafficking. Physiological substrates for these enzymes are thought to be regions of double-stranded RNA, because several DExH/D proteins catalyze strand separation in vitro. These "RNA helicases" can also disrupt RNA-protein interactions, but it is unclear whether this activity is coupled to duplex unwinding. Here we demonstrate that two unrelated DExH/D proteins catalyze protein displacement independently of duplex unwinding. Therefore, the essential functions of DExH/D proteins are not confined to RNA duplexes but can be exerted on a wide range of ribonucleoprotein substrates.     

Ventricular septal defect repair in a small dog using cross-circulation

SUMMARY A haemodynamically significant ventricular septal defect was diagnosed in a 3-month-old male Cavalier King Charles Spaniel. A median sternotomy was performed and the 6.5 kg dog placed on cardiopulmonary bypass using pump-assisted cross-circulation. A 10 mm diameter peri-membranous ventricular septal defect was closed using a continuous suture of 4–0 polypropylene, via a 2.5 cm incision in the right ventricular outflow tract. The duration of cardiopulmonary bypass was 90 minutes. Complications in the immediate postoperative period were mild and easily managed.     

Effect of CO2 enrichment on photosynthesis,growth and yield of tomato

Net photosynthesis of tomato plants was measured as CO₂ uptake in various light intensities, CO₂ concentrations, O₂ concentrations and temperatures during short-term experiments. Net photosynthesis increased significantly with increasing CO₂ concentration at all light intensities, even at the lowest one. The optimal temperature for photosynthesis increased with CO₂ enrichment. The changes in photosynthesis when the $\text{CO}{}_2\text{O}{}_2$ ratio was varied suggest that the effect of CO₂ enrichment is a result of a reduction in photorespiration.To determine whether the increase in photosynthesis caused by CO₂ enrichment would produce a greater yield, tomato plants were cultivated from seed to harvest in a tightly-closed greenhouse that was enriched with CO₂ continuously during the entire growing-period. The control greenhouse was ventilated by openings in the roof and door. The temperature of the 2 greenhouses was kept the same. Plants enriched with CO₂ showed a significant increase in fresh and dry weight and yield of tomatoes. The results are discussed in relation to earlier work in which CO₂ enrichment was discontinued when there was a need for ventilation.     

A novel IS element, ISMpa1, in Mycobacterium avium subsp. paratuberculosis

A novel insertion element belonging to the IS110 family was identified in Mycobacterium avium subsp. paratuberculosis. The IS element, ISMpa1, is 1500 bp and has one ORF encoding a putative transposase. Three copies of ISMpa1 were identified in the M. avium subsp. paratuberculosis genome. The element had inserted into the 3' end of the highly conserved mycobacterial genes prrB and a homologue of M. tuberculosis Rv1593c, and between a putative cytochrome p450 oxygenase and a putative hydrolase. The IS element was present in all (n = 11) M. avium subsp. paratuberculosis strains but not detected in most other mycobacterial species examined, including 10 M. avium subsp. avium isolates of human, avian and porcine origin. However two porcine isolates of M. avium subsp. avium and the reference strain IWGMT49 did harbour ISMpa1. These three strains belong to a previously described subgroup of M. avium subsp. avium based on IS1245 restriction fragment length polymorphism (RFLP) pattern and serovars. All of the M. avium subsp. paratuberculosis strains examined had an identical RFLP pattern when probed with sequences corresponding to the 5' end of ISMpa1, whereas a different pattern was seen in the positive M. avium subsp. avium strains. This novel IS element might be a useful tool in strain classification of M. avium subsp. avium and also for the identification of M. avium subsp. paratuberculosis when used in combination with IS900.     

Detection of Mycobacterium avium subsp. paratuberculosis by buoyant density centrifugation, sequence capture PCR and dot blot hybridisation

Detection of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) by polymerase chain reaction (PCR) is often hampered by the lack of efficient methods for sample treatment. We report a protocol for analysis of faecal samples based on buoyant density centrifugation in Percoll and IS900 sequence capture PCR combined with a dot blot assay for detection of low-grade infection of M. paratuberculosis. Serial dilutions of M. paratuberculosis genomic DNA and M. paratuberculosis bacteria were used to assess the sensitivity of the method. The final evaluation was performed with spiked faecal samples, which also were analysed by culture. The presence of PCR inhibitory substances in processed faecal samples was evaluated by including a PCR internal control. By using buoyant density centrifugation, sequence capture PCR, and dot blot hybridisation, we achieved a sensitivity of 10(3)CFU (colony forming units)/g of faeces. The detection limit by culture was assessed to 10(2)CFU/g of faeces. We conclude that the described protocol is a fast and sensitive alternative to bacterial culture of faecal samples.