Alkaloid binding and activation of D2 dopamine receptors in cell culture.

Ergot and pyrrolizidine alkaloids, either extracted from endophyte-infected tall fescue, synthesized, or purchased commercially, were evaluated in cultured cells to estimate their binding to the D2 dopamine receptor and subsequent effects on cyclic AMP production in GH4ZR7 cells, transfected with a rat D2 dopamine receptor. Ergopeptide alkaloid (alpha-ergocryptine, bromocryptine, ergotamine tartrate, and ergovaline) inhibition of the binding of the D2-specific radioligand, [3H]YM-09151-2, exhibited inhibition constants (K(I)) in the nanomolar range, whereas dopamine was less potent (micromolar). The lysergic acid amides (ergine and ergonovine) were 1/100th as potent as the ergopeptide alkaloids. Ergovaline and ergotamine tartrate were equally effective in inhibiting vasoactive intestinal peptide (VIP)-stimulated cyclic AMP production, with consistent nanomolar effective concentration (EC50) values. The remaining ergopeptide alkaloids (alpha-ergocryptine and bromocryptine), lysergic acid amides (ergonovine and ergine), and dopamine were 1/100th as potent. Two representative pyrrolizidines, N-formylloline and N-acetylloline, exhibited no binding activity at the D2 dopamine receptor or effects on the cyclic AMP system within the concentration ranges of nanomolar to millimolar. Our results indicate that the commercially available ergot alkaloids ergotamine tartrate and ergonovine may be used interchangeably in the D2 dopamine receptor system to simulate the effects of extracted ergovaline and ergine and to examine responses in receptor binding and the inhibition of cyclic AMP.     

Collaborative study of an improved method for the enumeration and confirmation of Clostridium perfringens in foods.

A collaborative study was conducted in 10 laboratories to evaluate the performance of a new method for the enumeration of vegetative cells of Clostridium perfringens in foods. Results obtained by the new method were compared with results from the official first action method, 46.049-46.053. Per cent recoveries of 4 C. perfringens strains from inoculated roast beef samples were higher and more consistent in tryptose-sulfite-cycloserine (TSC) agar with or without added egg yolk than in sulfite-polymyxin-sulfadiazine (SPS) agar, specified in the official first action method. The confirmatory technique utilized in the new method was also found to be more reliable than the technique described in the official first action method. Based on the collaborative results, the new method with TSC agar for enumeration and a modified motility-nitrate medium together with a lactose-gelatin medium for confirmation of C. perfringens has been adopted as official first action to replace 46.049-46.053.     

Plant-pest interactions in time and space: A Douglas-fir bark beetle outbreak as a case study

A conceptual model of Douglas-fir bark beetle (Dendroctonus pseudotsugae) dynamics and associated host tree mortality across multiple spatial and temporal scales was developed, then used to guide a study of the association between the occurrence of beetle- killed trees and factors that might render trees more susceptible to attack. Long-term records of beetle kill showed that beetle epidemics were associated with windstorms and drought at statewide and local spatial scales. At the landscape scale, beetle kill was associated with (i) portions of the landscape that were potentially drier (southern aspects, lower elevations) and (ii) portions of the landscape that had more mature and old-growth conifer vegetation. The patches of beetle-killed trees were aggregated with respect to other patches at scales of approximately 1 and 4 km. At the scale of the individual tree, there was not a strong relationship between beetle kill and resistance to attack measured by tree growth rate prior to attack. Our results show that landscape-scale phenomena and temporal patterns were more strongly correlated with beetle-kill events than was recent growth history at the scale of individual trees. We suggest that the multi-scale approach we employed is useful for elucidating the relative roles of fine- versus coarse-scale constraints on ecological processes.     

Changes in condition factor and gonadosomatic index in maturing and non-maturing Atlantic salmon (Salmo salar L.) in Bay of Fundy sea cages, and the effectiveness of photoperiod manipulation in reducing early maturation

Changes in condition factor (cf) and gonadosomatic index (GSI) in maturing and non‐maturing Atlantic salmon (Salmo salar L.) cultured in Bay of Fundy sea cages was investigated under control (natural) and continuous light conditions. All experimental cages contained salmon stocked as smolts in April 2001. Six control cages received only natural illumination, three received continuous light from 3 November, 2001 until 31 May, 2002, and three received continuous illumination from 15 February, 2001 until 31 May, 2002. In mid‐July 2002, each cage was sampled, and sex, round weight, fork length (FL), mean muscle lipid content and gonad weight were recorded from each sampled fish. At harvest (August 2002 to February 2003), sex, maturation state, round weight and FL were measured from a sample of fish from each cage. The frequency distributions of GSIs from fish sampled in July indicated for both sexes a GSI>0.3 was indicative of early maturation. The relationship between cf and GSI suggested that salmon of both sexes had to have a cf greater than 1.3 in early summer for early maturation to develop. Continuous illumination from November greatly reduced the number of males with GSI>0.3 in mid‐July (0.8%, compared with 50% for control males); 17% of males from cages lit in February had GSIs exceeding 0.3. The percentage of control females exceeding 0.3 was 7%, compared with 0% and 4% for females from cages receiving continuous illumination from November and February respectively. For salmon with GSI of 0.3 or less, the percent muscle lipid increased linearly with cf. A multiple regression of cf and FL on GSI (GSI=?0.973+0.35 cf+0.013 FL) correctly identified 88% of maturing vs. non‐maturing salmon from control cages in July.     

Influence of incubation temperature on body movements of Atlantic cod (Gadus morhua L.) embryos and on size at hatch

Body movements of cod (Gadus morhua L.) embryos reared from fertilization to hatch at 5.4°C were observed at various stages of development and at six experimental temperatures ranging from 0–10°C. Frequency of cod embryo body movements increased from zero at 42 degree‐days post fertilization to maximal at 73–82 degree‐days (1 or 2 days prior to hatch). Embryos were most active at 2°C (mean of 5.5 movements per 10 min), with activity declining to less than 1/10 min at 8–10°C. Lengths of hatched cod larvae reared at a series of constant temperatures (from 4–10°C) from fertilization to hatch were greater at lower incubation temperatures. Incubation temperatures of 2–4°C were found to be optimal for incubation of cod eggs.     

Styles of volcanism on venus: new arecibo high resolution radar data

Arecibo high-resolution (1.5 to 2 km) radar data of Venus for the area extending from Beta Regio to western Eisila Regio provide strong evidence that the mountains in Beta and Eisila Regiones and plains in and adjacent to Guinevere Planitia are of volcanic origin. Recognized styles of volcanism include large volcanic edifices on the Beta and Eisila rises related to regional structural trends, plains with multiple source vents and a mottled appearance due to the ponding of volcanic flows, and plains with bright features surrounded by extensive quasi-circular radar-dark halos. The high density of volcanic vents in the plains suggests that heat loss by abundant and widely distributed plains volcanism may be more significant than previously recognized. The low density of impact craters greater than 15 km in diameter in this region compared to the average density for the higher northern latitudes suggests that the plains have a younger age.     

Glucose-stimulated lipolysis in rainbow trout,Oncorhynchus mykiss,liver

Rainbow trout, Oncorhynchus mykiss, were used to characterize further the influence of glucose on hepatic lipolysis. Liver was removed from fed fish, cut into 1 mm³ pieces and incubated for up to 5 h in Hanks medium containing either 2 mM, 5.5 mM, 10 mM, or 25 mM glucose. Glucose-stimulated lipolysis was indicated by tissue triacylglycerol (TG) lipase activity and by medium concentrations of glycerol and fatty acids (FA). Triacylglycerol lipase activity in liver pieces incubated in the presence of higher concentrations (25 mM) of glucose was significantly higher than that in liver pieces incubated in lower concentrations (2 mM) of glucose, rising from 0.075 ± 0.002 (mean ± SEM) nmol FA released/h/mg protein to 0.092 ± 0.004 units. Similarly, higher concentrations of glucose stimulated significantly more FA release and glycerol release from liver pieces than that stimulated by lower concentrations of glucose. Glycerol release from liver pieces incubated in the presence of 10 mM glucose and 25 mM glucose was ca. 2-fold to 2.8-fold, respectively, higher than that from liver pieces incubated in the presence of either 2 mM or 5.5 mM glucose. Fatty acid release from liver pieces incubated in the presence of 10 mM or 25 mM glucose was ca. 1.8-fold higher than that from liver pieces incubated in the presence of either 2 mM or 5.5 mM glucose. Notably, increased glycerol release was not accompanied by a parallel increase in FA. Fatty acid reesterification was more pronounced in liver pieces exposed to higher glucose (10 mM and 25 mM) than in liver pieces exposed to lower glucose (2 mM and 5.5 mM). ¹⁴C-incorporation studies indicated that glucose serves as a carbon source for reesterified FA in trout liver. The route of reesterification appears to be from glucose to glycerophosphate to phosphatidic acid to diacylglycerol to TG. Increasing concentrations of glucose did not affect glycerol kinase activity, indicating that glucose-stimulated lipolysis was not accompanied by increased glycerol recycling within the liver. These results suggest that glucose stimulates fatty acid reesterification and directly enhances net lipolysis in trout liver incubated in vitro.A part of this study was presented at the Annual Meeting of the American Society of Zoologists, December 26–30, 1991, Atlanta, GA.     

Isolation of bovine herpesvirus type 5 from the semen of a healthy bull in Australia

Artificial insemination is widely used in the cattle industry and a major challenge is to ensure that semen is free of infectious agents. A healthy donor bull was tested for freedom from infectious agents. A bovine herpesvirus was isolated in testis cells and identified as bovine herpesvirus type 5 (BoHV-5) by polymerase chain reaction and by direct amplicon sequencing. The amplicon sequence shared 100% similarity with the published sequence of BoHV-5. This is the first report in Australia of BoHV-5 in semen. The implications of this finding are discussed.     

Growth Hormone Receptors in the Porcine Testis During Prepuberty

Supplementation of exogenous growth hormone (GH) during prepuberty advances onset of spermatogenesis in boars, but the mechanism of action is unknown. The present study is an investigation of the presence and characteristics of testicular growth hormone receptors (GHR). A total of 36 boars were castrated, three boars every 10 days, between the ages of 10 and 120 days. Testicular membrane preparations of 10, 20, 30, 50, 70, 100 and 120‐day‐old boars were used to determine ¹²⁵I‐bGH binding and Scatchard analysis. Liver from a 60‐kg barrow was used for comparison. Specific ¹²⁵I‐bGH binding to testicular membrane preparations occurred in all age groups with the exception of 20‐day‐old boars at levels of 30–40% of liver binding. At 30 days of age the unlabelled bGH at 1.1 ng/tube achieved half maximal inhibition (ID₅₀). Results of Scatchard analysis indicated a single class of binding sites. Binding affinity was 2.89 × 10⁹m with a binding capacity of 12 fmole/mg membrane protein. The results from this study suggest that GH may act directly on the cells of the prepubertal boar testis.