Purification and characterization of hepatic triacylglycerol lipase isolated from rainbow trout,Oncorhynchus mykiss

Trialcylglycerol (TG) lipase was isolated and partially purified from rainbow trout liver. Triacylglycerol lipase activity was assayed by measuring¹⁴C-oleic acid release from¹⁴C-triolein.¹⁴C-oleic acid release was linear for up to two hours. Optimal activity occurred at pH 7.0 and 15°C. Most of the lipase activity was recovered in the cytosolic fraction. A 27,000-fold purification was achieved after Sepharose (Bio-gel A 0.5 M, 200–400 mesh) chromatography of a resuspended 20% ammonium sulfate fraction. The molecular weight of the trout hepatic lipase as determined by size-exclusion chromatography and by SDS-polyacrylamide gel electrophoresis was 40–43 kD. Lipase-mediated hydrolysis of TG resulted in the production of diacylglycerols, monoacylglycerols, and fatty acids. Kinetic analysis indicated that V_max=0.016 nmol/h/mg protein and that K_m=0.28 mM triolein. Lipolytic activity was enhanced in the presence of cAMP/ATP-Mg²⁺. These results suggest that the liver of trout possesses a neutral TG lipase that is responsible for mobilizing stored TG and is catalytically activated by phosphorylation.A part of this work was presented at the Annual Meeting of the American Society of Zoologists, December 26–30, 1990, San Antonio, TX.     

Ergovaline does not alter the severity of ryegrass staggers induced by lolitrem B

AIM: To investigate a possible interaction between lolitrem B and ergovaline by comparing the incidence and severity of ryegrass staggers in sheep grazing ryegrass (Lolium perenne) containing lolitrem B or ryegrass containing both lolitrem B and ergovaline.

METHODS: Ninety lambs, aged approximately 6 months, were grazed on plots of perennial ryegrass infected with either AR98 endophyte (containing lolitrem B), standard endophyte (containing lolitrem B and ergovaline) or no endophyte, for up to 42 days from 2 February 2010. Ten lambs were grazed on three replicate plots per cultivar. Herbage samples were collected for alkaloid analysis and lambs were scored for ryegrass staggers (scores from 0–5) weekly during the study. Any animal which was scored ≥4 was removed from the study.

RESULTS: Concentrations of lolitrem B did not differ between AR98 and standard endophyte-infected pastures during the study period (p=0.26), and ergovaline was present only in standard endophyte pastures. Ryegrass staggers was observed in sheep grazing both the AR98 and standard endophyte plots, with median scores increasing in the third week of the study. Prior to the end of the 42-day grazing period, 22 and 17 animals were removed from the standard endophyte and AR98 plots, respectively, because their staggers scores were ≥4. The cumulative probability of lambs having scores ≥4 did not differ between animals grazing the two pasture types (p=0.41).

CONCLUSIONS AND CLINICAL RELEVANCE: There was no evidence for ergovaline increasing the severity of ryegrass staggers induced by lolitrem B. In situations where the severity of ryegrass staggers appears to be greater than that predicted on the basis of concentrations of lolitrem B, the presence of other tremorgenic alkaloids should be investigated.     

The apparent prevalence of skin lesions suspected to be human-inflicted in Danish finishing pigs at slaughter

Skin lesions on pigs inflicted by humans compromise animal welfare and are the subject of increased public and political attention in Denmark. Systematic surveillance of such skin lesions was enforced in April 2010 at all Danish pig abattoirs, through the recording of meat inspection Code 904 for the presence of skin lesions suspected to be human inflicted. The objectives of the present study were to (a) estimate the apparent prevalence of Code 904s at the pig and herd owner level; (b) characterise the distribution of deliveries with pigs demonstrating a Code 904; (c) characterise the distribution of herd owners with repeated Code 904 recordings; and (d) determine the developments in Code 904 prevalence over time in Danish finishing pigs in the period from May 1, 2010 to September 30, 2013.     

Carbon stocks, soil respiration and microbial biomass in fire-prone tropical grassland, woodland and forest ecosystems

A thorough understanding of the role of microbes in C cycling in relation to fire is important for estimation of C emissions and for development of guidelines for sustainable management of dry ecosystems. We investigated the seasonal changes and spatial distribution of soil total, dissolved organic C (DOC) and microbial biomass C during 18 months, quantified the soil CO₂ emission in the beginning of the rainy season, and related these variables to the fire frequency in important dry vegetation types grassland, woodland and dry forest in Ethiopia. The soil C isotope ratios (δ¹³C) reflected the 15-fold decrease in the grass biomass along the vegetation gradient and the 12-fold increase in woody biomass in the opposite direction. Changes in δ¹³C down the soil profiles also suggested that in two of the grass-dominated sites woody plants were more frequent in the past. The soil C stock ranged from being 2.5 (dry forest) to 48 times (grassland) higher than the C stock in the aboveground plant biomass. The influence of fire in frequently burnt wooded grassland was evident as an unchanged or increasing total C content down the soil profile. DOC and microbial biomass measured with the fumigation-extraction method (C_mic) reflected the vertical distribution of soil organic matter (SOM). However, although SOM was stable throughout the year, seasonal fluctuations in C_mic and substrate-induced respiration (SIR) were large. In woodland and woodland-wooded grassland C_mic and SIR increased in the dry season, and gradually decreased during the following rainy season, confirming previous suggestions that microbes may play an important role in nutrient retention in the dry season. However, in dry forest and two wooded grasslands C_mic and SIR was stable throughout the rainy season, or even increased in this period, which could lead to enhanced competition with plants for nutrients. Both the range and the seasonal changes in soil microbial biomass C in dry tropical ecosystems may be wider than previously assumed. Neither SIR nor C_mic were good predictors of in situ soil respiration. The soil respiration was relatively high in infrequently burnt forest and woodland, while frequently burnt grasslands had lower rates, presumably because most C is released through dry season burning and not through decomposition in fire-prone systems. Shifts in the relative importance of the two pathways for C release from organic matter may have strong implications for C and nutrient cycling in seasonally dry tropical ecosystems.     

Akabane disease in sheep

Perinatal lamb mortality, associated with malformations of the CNS due to Akabane viral infection, occurred in 4 of 9 flocks of ewes lambing on 3 farms between 26 May and 14 November, 1976. Cases were restricted to ewes conceiving prior to the second week of March and lambing between 26 May and 19 July. As judged by seroconversion in sentinel flocks on 2 of the farms, field infection with Akabane virus occurred mainly between mid-February and mid-April. Malformations of the CNS occurred in 42.5%, 51.2%, 100% and 31.0% of the dead lambs examined in the affected flocks respectively. Prevalence in the 4 affected flocks, expressed as the proportion of ewes lambing which delivered at least one malformed foetus, was 6.1%, 8.4%, 88.9% and 5.7% respectively. Lamb mortality due to malformations of the CNS was 7.1%, 5.5%, 92.3% and 5.7% of lambs born. Age-specific prevalence was calculated for 3 of the 4 flocks and 2-year-old ewes accounted for 71.4% and 76.9% of total cases respectively in 2 flocks, whereas in one flock malformations occurred at equivalent frequencies throughout several older age groups. Birthweights of affected lambs were usually significantly lighter than those of unaffected lambs of similar sex and birth-type, and their mean duration of gestation was slightly, and significantly, prolonged. Micrencephaly (88.1% of cases) and hydrocephalus (68.7% of cases) were the outstanding pathological features of the malformations with hydranencephaly, microgyria, porencephaly and attenuation of the spinal cord occurring at much lower frequencies.(ABSTRACT TRUNCATED AT 250 WORDS)     

Moderate nitrogen retention in temperate heath ecosystem after elevated CO2, drought and warming through 7 years

Nitrogen (N) dynamic is one of the main controlling factors of responses to climate change in N-limited terrestrial ecosystems, which rely on nutrient recycling and retention. In this study we investigate the N partitioning in ecosystem compartments of a grassland heath, and the impact of multiple climate change factors on long-term N retention after ¹⁵N pulse labelling. The impacts of elevated carbon dioxide (eCO₂), warming and drought and the treatments in combination on ecosystem N retention were investigated in a field scale manipulation experiment. A 6-year time-course was assessed by pulse-labelling with the stable N isotope ¹⁵N and by sampling after 1 day, 1 year and 6 years. After 6 years we observed that the total ecosystem retained 42% of the amended ¹⁵N across treatments (recovery of the amended ¹⁵N in the pool). The fate of the applied ¹⁵N was mainly stabilization in soil, with 36% ¹⁵N recovery in soil, while the plant compartment and microbial biomass each retained only 1%–2% of the added ¹⁵N. This suggests a moderate retention of N, for all treatments, as compared to similar long-term studies of forest ecosystems. A decreased ammonium and vegetation N pool combined with higher ¹⁵N retention in the soil under eCO₂ treatments suggests that eCO₂ promoted processes that immobilize N in soil, while warming counteracted this when combined with eCO₂. Drought treatments contrastingly increased the vegetation N pool. We conclude that as the organic soil layer has the main capacity for N storage in a temperate heathland-grassland, it is important for buffering nutrient availability and maintaining a resilient ecosystem. However, the full treatment combination of drought, warming and eCO₂ did not differ in ¹⁵N recovery from the control, suggesting unchanged long-term consequences of climate change on retention of pulse added N in this ecosystem.