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.     

Maximal lactate concentrations in horses after exercise of different duration and intensity

Lactate kinetics in whole blood of horses was investigated after exercise of differing velocities and duration. The following categories of exercise were used: A: <11 m/second and >180 seconds (n=35), B: >11 m/second and <180 seconds (n=17) and C: <11 m/second and <180 s (n=10). The mean peak lactate concentration determined in horses in category A was 4.49 ± 2.21 mmol/1, in B, 16.32 ± 4.81 mmoVl and in C, 4.58 ± 1.59 mmol/l. While the maximum lactate concentrations in categories A and C were always found immediately after the exercise, the peaks in category B were measured between the first and tenth minute after exercise. Mean lactate concentrations measured at 2-minute intervals after bouts of category-B exercise tended to stabilize 3 to 10 minutes after exercise; however, mean lactate concentrations measured during the intervals before and after the peak value differed significantly. The lactate concentration returned to pre-exercise levels within 20 minutes after exercise bouts of category C, but remained above pre-exercise levels up to 60 minutes after bouts of category-A and -B exercise. It was concluded that, for an evaluation of lactate data after intensive anaerobic exercise, sequential blood sampling at 2-minute intervals for a period of up to 12 minutes after exercise is necessary. Less frequent sampling may be a reason for the often described irreproducibility of lactate concentrations in horses. After aerobic or mild anaerobic exercise, one sample is sufficient, but it has to be taken as soon as possible after exercise.     

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.     

Variation of the growth hormone gene in ryr 1 genotyped Austrian pig breeds

SUMMARY: Polymorphism in the second intron of the porcine growth hormone gene of 273 Austrian Landrace and 81 Austrian Edelschwein pigs was investigated with a PCR-RFLP-technique. Results showed significantly different genotype patterns between the two breeds. The frequency of the Hae II(-) allele was significantly (P < 0,001) higher in the landrace than in the Edelschwein population (0,83 and 0,47 resp.). The Msp I(+) allele was predominant in both breeds but signifanctly higher in the Landrace (0,98 versus 0,69; P < 0,01). Analyses the Hae II/Msp I locus combination revealed also in breed specific difference. In the Landrace a very low interaction was found between the Hae II and ryr 1 locus, and between Msp I and ryr 1 locus (c. c. = 0,181 and 0,186 resp.). The correlation was slightly stronger (c. c. = 0,266) between the ryr 1 and Hae II/Msp I genotypes. No correlation was detected among the three loci in the Edelschwein population. ZUSAMMENFASSUNG: Variabilit?t des Somatotropin Gens in ?sterreichischen Schweinerassen genotypisiert hinsicbtlich ryr 1 An 273 ?sterreichischen Landrasse Schweinen und 81 ?sterreichischen Edelschweinen wurde der Polymorphismus am zweiten Intron des Schweinewachstumshormons mittels PCR-RFLP-Technik untersucht. Genotypen-und Genfrequenzen waren zwischen den beiden Rassen signifikant verschie- den. Die Hae II(-) Allelfrequenz war bei den Landrassetieren signifikant h?her (P < 0,001) als bei den Edelschweinen (0,83 bzw. 0,47). In beiden Rassen überwiegte das Msp I(+) Allel, das aber signifikant ?fters bei der Landrasse auftrat (0,98 bzw. 0,69; P < 0,01). Die Verteilung der Locuskombination von Hae II/Msp I zwischen den beiden Rassen war ebenfalls unterschiedlich. Bei der Landrasse konnte nur ein sehr geringer Zusammenhang zwischen dem Hae II und ryr 1 Locus (c. c. = 0,181 bzw. 0,186) und auch zwischen dem Msp I und ryr 1 Locus festgestellt werden. Die Korrelation zwischen dem ryr 1 und Hae II/Msp I Genotyp war geringfügig gr??;er (c. c. = 0,266). Bei den Edelschweinen konnte kein Zusammenhang zwischen den drei untersuchten Loci festgestellt werden.     

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.     

Contribution to the taxonomy and pathogenicity of fungicolous Verticillium species. II. Pathogenicity

In recent years in the Netherlands a second mushroom species,Agaricus bitorquis, which prefers higher temperatures thanA. bisporus and is less susceptible to certain diseases, is often commercially grown.Verticillium fungicola var.fungicola, the causal agent of dry bubble, is responsible for considerable damage in crops ofA. bisporus. InA. bitorquis, however, dry bubble has hardly been noticed, but brown spots due toV. fungicola var.aleophilum resulted in inferior mushroom quality. The latter variety also caused brown spots ina. bisporus, but to a minor degree. In variety Les Miz 60 ofA. bisporus, however, it also induced fruit-body deformation in a way different from dry bubble. Verticillium psalliotae, isolated fromA. bitorquis in England, induced more confluent brown spots inA. bitorquis. In the netherlands, where moreA. bitorquis is grown than in other countries,V. psalliotae has not yet been encountered in crops ofA. bitorquis. V. psalliotae, which has a high temperature optimum for mycelial growth, likeV. fungicola var.aleophilum andA. bitorquis, did not infectA. bisporus in our trials.Artificial infection ofA. bisporus orA. bitorquis could not be accomplished with the following related and/or fungicolous fungi:Verticillium lamellicola, V. fungicola var.flavidum, V. biguttatum, Nectriopsis tubariicola, Acremonium crotocinigenum andAphanocladium album.Samenvatting Vooral in Nederland wordt sinds een aantal jaren naastAgaricus bisporus ook de warmteminnende champignonsoortAgaricus bitorquis geteeld, die minder vatbaar is voor bepaalde ziekten. TerwijlVerticillium fungicola varfungicola in de teelt vanA. bisporus droge mollen en daardoor veel schade veroorzaakt, komen in de teelt vanA. bitorquis geen droge mollen voor maar wel bruine vlekken, die tot kwaliteitsverlies en dus schade leiden. De vlekken bleken veroorzaakt te worden doorV. fungicola var.aleophilum. Deze schimmel veroorzaakte ook inA. bisporus bruine vlekken, hoewel in minder ernstige mate, maar in het ras Les Miz 60 vanA. bisporus bovendien misvormde champignons, die wel op droge mollen leken, maar daaraan niet gelijk waren.OokV. psalliotae, in Engeland geïsoleerd vanA. bitorquis met vlekken, veroorzaakte wat meer samenvloeiende, bruine vlekken inA. bitorquis. In Nederland, waar meerA. bitorquis geteeld wordt dan in andere landen, isV. psalliotae nog niet aangetroffen in teelten vanA. bitorquis. InA. bisporus kon geen kunstmatige infectie worden verkregen metV. psalliotae, die net alsV. fungicola var.aleophilum enA. bitorquis warmteminnend genoemd zou kunnen worden.Met de volgendeVerticillium-achtige of van paddestoelen geïsoleerde schimmels kon evenmin op kunstmatige wijze een infectie worden opgeroepen inA. bisporus ofA. bitorquis: Verticillium lamellicola, V. fungicola var.flavidum, V. biguttatum, Nectriopsis tubariicola, Acremonium crotocinigenum enAphanocladium album.