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.     

Waldschutz gegen Borkenkäfer: Der Beitrag von Duftstoffmeteorologie und Populationsdynamik

Zusammenfassung Mit theoretischen Modellen kann die Effektivität verschiedener Ansätze (z. B. Fallenzahl und-abstand, Ködermenge und-abgaberate) für den Einsatz von synthetischen Pheromonen in Pest-Management-Systemen verglichen werden. Das GAUSS-sche Modell, das von einer Normalverteilung der Konzentrationsabnahme um eine Lockstoffquelle ausgeht, wurde benutzt, um fürT. lineatum die lockwirksame Reichweite und die Konkurrenzsituation von Naturköder und Falle abzuschätzen. Bei 100 000 /ha und 2 Wochen Flugdauer ergeben sich für (+)-Lineatin (Reaktionsschwelle 0,1 ng/m³, Produktion 6,43 ng//d, Produktionsdauer 3 d/) mittlere Tagesraten von 128,6 g/ha bzw. eine Gesamtproduktion von 1,929 mg/ha. Die lockwirksame Reichweite x_max beträgt je nach Wetterlag für 1T. lineatum- 20–40 cm, für Fallen mit 1 CONREL-Faser (Abgaberate 10 g/d [+]-Lineatin) 13–16m. Für eine 10fach höhere lockwirksame Reichweite sind nach dem Modell etwa 100fach, in der Praxis nach verschiedenen Autoren sogar 200fach stärkere Köder erforderlich.In Freilandversuchen über die Relation von Fangleistung und Beifängen unterschiedlicher Fangsysteme waren die Coleoptera (bes. Rhizophagidae und Staphylinidae) am häufigsten. In Trichterfallen lagen die Beifänge mit 9,2% deutlich höher als in Schlitzfallen (1,2%). Das Giftpolter hatte demgegenüber das größte Beifangspektrum; hier traten auch Ordnungen auf, die von anderen Fangsystemen nicht betroffen waren.Massenfang kan prinzipiell in Holzhöfen und Waldbeständen erfolgreich durchgeführt werden. Allerdings ist das Problem der Beifänge noch ungelöst und die Relations von potentieller Vermehrung der Borkenkäfer und maximal möglicher Populationsreduzierung durch Fallenfang unbekannt. Zudem kann eine nach Massenfang geringere Populationsdichte im Folgciahr durch zunehmende Reproduktionsraten oder durch ersatzweise Einnischung (competive displacement) anderer Borkenkäferarten ausgeglichen und damit der Forstschutzeffekt zunichte gemacht werden.

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Bark beetle control: contribution of pheromone meteorology and population dynamics

The use of semiochemicals in pest management systems of bark beetles is viewed in relation to optimization of resources investment, comparing beetle numbers in univoltine, requisitegoverned woodboringTrypodendron lineatum (Oliv.) with multigeneration bark inhabitingIps typographus (L.). The efficiency of different numbers of and distance between traps and pheromone load of dispensers was analysed using steady state models and comparing results with published and own field data. Competition between traps and beetle-infested logs, attractive distance and overlap of pheromone plume was compared forT. lineatum using models which assume a GAUSS distribution of concentration decrease around a pheromone source. For (+)-lineatin (reaction threshold 0.1 ng/m³, production 6.43 ng//d, duration of production 3 d/), 100,000 /ha and a flight period of 2 weeks, daily mean production is 128.6 g/ha. Attractive distance x_max is 20–40 cm for 1T. lineatum- and 13–16 m for a trap with 1 CONREL-dispenser (mean release rate 10 g/d (+)-lineatin). A 10-fold increase of attractive distance x_max needs a 100-fold increase of pheromone release rate according to the model, and 200-fold according to field experiments.In field experiments, Coleoptera (esp. Staphylinidae and Rhizophagidae) were the most abundant non-target trap catches. Insecticide-treated trap logs had the widest range of non-target insects including orders that were not affected by the non-insecticide trapping systems. Mass trapping can be successful in logging areas and/or forests. However, the exclusion of non-target insects still is an unsolved problem and we still have only limited knowledge on potential population increase of beetles and maximum reduction that can be achieved by trapping. Also, population levels reduced by mass trapping can be compensated by increasing reproductive effort of the succeeding generation and/or competitive displacement by secondary bark beetle species.

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Mit 5 Abbildungen und 2 Tabellen

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Mit Unterstützung der Deutschen Forschungsgemeinschaft.

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Nach einem Vortag bei der Fortbildungstagung des Hessischen Forstvereins in Solms/Lahn am 23. Mai 1991.

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Herrn Prof. Dr. Dr.G. Wellenstein zum 85. Geburtstag.     

Direct electron microscopy and serology with plant viruses in leaf material dried and stored over calciumchloride

Most of 43 viruses could easily be detected directly in 53 out of 66 leaf samples dried and stored over CaCl₂ for varying periods of time up to 20 1/2 years. Detection usually was with PTA pH 6.5, but alfalfa mosaic, cucumber mosaic and tomato aspermy viruses required PTA pH 3.0 to 4.0. Bean common mosaic, cowpea aphid-borne mosaic and cowpea mosaic viruses were also easily observed in newly dehydrated samples obtained for diagnosis from Morocco and Tanzania.Broad bean wilt virus, cowpea mosaic virus and cucumber mosaic virus were detected with agar gel-diffusion tests in dry leaf material ground in buffer. This serological assay demonstrated a high concentration of cucumber mosaic virus in leaf material dried over CaCl₂ 20 years ago. This paper further coroborates the value of the CaCl₂ method of dehydration and storage of plant viruses in leaf material.Samenvatting Zeer uiteenlopende plantevirussen kunnen goed worden bewaard in bladmateriaal van geïnfecteerde planten dat is gedroogd en bewaard boven CaCl₂. Bij rechtstreekse elektronenmicroscopische toetsing van 66 monsters met verschillende isolaten van 43 virussen konden in 53 monsters gemakkelijk virusdeeltjes worden waargenomen (Tabel 1 en Fig. 1 en 2). Meestal lukte dit met fosforwolfraamzuur pH 6,5 dat doorgaans voor in ruw plantesap voorkomende virussen wordt gebruikt. Bij luzernemozaïekvirus en komkommermozaïek virus gelukte dit alleen maar bij lagere pH (3,0 en 4,0). Ook het tomate-aspermievirus was dan veel gemakkelijker aantoombaar. Methylaminewolfraamzuur gaf geen beter resultaat.Draden van bonerolmozaïekvirus en van cowpea aphid-borne mosaic virus en bolletjes van cowpea mosaic virus waren snel waarneembaar in elektronenmicroscopische preparaten gemaakt van bladmateriaal van recent uit Marokko en Tanzania ontvangen monsters.Cowpea mosaic virus, tuinboneverwelkingsvirus en komkommermozaïekvirus konden eveneens gemakkelijk en snel serologisch worden aangetoond in met bufferoplossing vermalen droog blad van respectievelijk cowpea, erwt en tabak. Het laatstgenoemde, reeds 20 1/2 jaar geleden gedroogde bladmateriaal, bleek in vergelijking met pas geïnoculeerd vers blad van komkommer enChenopodium quinoa zelfs zeer veel serologisch actief virusmateriaal te bevatten (Fig. 3).De beschreven waarnemingen bevestigen nogmaals de waarde van de toegepaste methode van virusbewaring en tonen aan dat het mogelijk is vele virussen te herkennen in van elders ontvangen gedroogde bladmonsters, zonder het risico te lopen van virusontsnapping, zoals altijd aanwezig bij werk met toetsplanten in de kas.Guestworker from Istituto Sperimentale per la Patologia Vegetale, Rome, Italy, participating in the investigations from September 21 to December 21, 1977.     

Some currently available insecticides and their comparative efficacy on louse-infested, long-woolled sheep

The comparative efficacy of 13 of the sheep dips currently registered in New Zealand was investigated using sheep infested with the louse Bovicola ovis and carrying wool which was about 10 cm long at the shoulder. With the exception of one synthetic pyrethroid pour-on formulation, all products were able to effect a significant reduction in louse populations, relative to untreated controls, for 37 days after treatment. Only four products proved capable of eradicating lice and preventing their re-establishment up to 37 days after treatment. Variations in manufacturers' recommendations relating to the length of wool at dipping, and mode of application of dips are discussed in relation to the results.     

An evaluation of two cypermethrin-based pour-on formulations on sheep infested with the biting louse, Bovicola ovis

Two synthetic pyrethroid (cypermethrin) based pour-on insecticide formulations, with high cis/trans isomer ratios (80:20) but differing in their respective active ingredient concentrations and solvent component(s), were applied to sheep infested with the biting-louse, Bovicola ovis. All treated sheep were penned with louse-infested sheep 9,12 and 15 weeks after the insecticide was applied. The 2% cypermethrin formulation achieved a higher level of control than the 1.25% cypermethrin formulation at each challenge interval when applied 12 weeks after shearing. The 2% cypermethrin formulation provided 97-100% control of lice from 4 to 16 weeks after application on sheep shorn 6 or 12 weeks prior to treatment. The 1.25% cypermethrin formulation provided 85% control of lice 4 weeks after application on sheep shorn 12 weeks prior to treatment, the level of control increasing to a maximum of 100% by week 9, and declining thereafter. The 2% cypermethrin formulation may provide a better level of control in long-woolled sheep than 1.25% cypermethrin, by compensating for the diluent effect of lipid.