Differential winter mortality and seasonal selection in the polymorphic ladybird Adalia bipunctata (L) in the Netherlands

Seasonal selection acting on the melanic polymorphism in the two-spot ladybird Adalia bipunctata was investigated in The Netherlands. An increase in melanic frequency over the spring-summer reproductive period was quantified. The selective advantage gained by melanics averaged 9%, but significant heterogeneity occurred between populations. Adult hibernation behaviour is described. The beetles when outdoors show a highly clumped distribution both between and within trees. The distribution of the morph classes between aggregations is random. Survivorship in a hibernating cohort (initial n= 1898) on a grid of 70 lime trees near Utrecht was monitored by making three counts over the winter of 1981–1982. Intense selection favouring each melanic morph occurred during December and January. The relative fitness of non-melanics was 0.55 (melanics =1). The discovery of dead beetles in late January (about 5% of total losses) and the absence of spatially density-dependent mortality were consistent with a climatic stress rather than selective predation. The period of selection was associated with very cold temperatures averaging up to 4°C below normal and an overall mortality of nearly 75%. There was no change in morph frequency, near normal temperatures and a lower mortality from February to early April. Examination of groups of nearby trees in late January strongly suggested that similar differential mortality had occurred except on some willows. This difference was probably due to the more protected hibernation sites available on these trees. Samples of hibernating cohorts at three other sites showed no evidence of differential mortality. Laboratory experiments with hibernating beetles found no difference in survivorship or rate of weight loss between starved non-melanics and melanics in temperature regimes with and without periods of adult activity. It is concluded that the intense winter selection on the study limes is probably exceptional. Examination of changes in morph frequency through the annual cycle suggests that at some sites the selection favouring melanics during reproduction is counterbalanced by selection against melanics in late summer or early autumn. The results are discussed in relation to mathematical models of cyclical selection and to other field studies including that of Timoféeff-Ressovsky (1940), who found large decreases in melanic frequency during hibernation in Berlin.     

Oviposition deterring pheromone in Rhagoletis cerasi: Behavioral laboratory test to measure pheromone activity

Current investigations concerning the identification of the chemical nature of the oviposition deterring pheromone of the European cherry fruit fly, Rhagoletis cerasi L., are conducted in an interdisciplinary research program by combining chemistry with behavioral evaluation. The methods used to collect and evaluate the pheromone by an improved semi-quantitative behavior test are described.

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Zusammenfassung Nach einer einleitenden Übersicht über den heutigen Stand der Erkenntnisse auf dem Gebiet der eiablage-hemmenden Pheromone (ODP) bei Fruchtfliegen und insbesondere der Kirschenfliege wird der verbesserte Verhaltenstest beschrieben, welcher für den halb-quantitativen Nachweis von ODP-Aktivität entwickelt worden ist. Im laufenden interdisziplinären Forschungsprogramm an der Eidg. Forschungsanstalt Wädenswil wird versucht, durch enge Zusammenarbeit zwischen Biologe, Chemiker und Elektrophysiologe das ODP der Kirschenfliege möglichst mit geringem Anteil an Verunreinigung einzusammeln, zu reinigen und einer chemischen Identifikation der ODP Komponenten zugänglich zu machen.Die Möglichkeiten und Grenzen des Verhaltenstests werden diskutiert und die Empfindlichkeit des Verfahrens mittels einer Konzentrationsreihe von Pheromonextrakt aufgezeigt.

     

Aspects of the numerical and functional respones of the aphid parasite, Aphidius sonchi, in the laboratory

The fecundity, reproductive rates, and adult survival of Aphidius sonchi Marshall (Hymenoptera: Aphidiidae) parasitizing second and third instar nymphs of the sowthistle aphid, Hyperomyzus lactucae (L.) (Homoptera: Aphididae) were measured at six different host densities under constant laboratory conditions. At host densities of less than 50 aphids per flowering shoot per female per day, oviposition constraints resulting from the lack of hosts reduced the number of eggs laid, enhanced the extent of superparasitization and, as a result, effectively lowered the fecundity and reproductive rates of the parasites. Above this host density the parasites laid on average 220–230 eggs, but the effective fecundity and reproductive rates continued to increase with the host density. By contrast, the survivorship of the parasites seemed unaffected by host density, with an average adult life span of 4–5 days at all densities. Analysis of the data showed that the intrinsic rate of increase (r_m) of the parasite varied with the host density and could reach values higher than that of the host under identical conditions. The response of r_m to changes in host density and parasite sex ratio is illustrated.Overall, A. sonchi showed a typical convex functional response, to host density. However, the response showed obvious changes through the parasite's adult life and, furthermore, the rates of changes were not consistent at all host densities. The frequency distributions of parasite eggs were generally indistinguishable from random, and the number of hosts parasitized were predicted satisfactorily by the random oviposition equation.

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Résumé L'étude a porté sure l'influence de 6 densités différentes d'Hyperomyzus lactucae (L.) (Homoptera: Aphididae), en conditions constantes de laboratoire, sur la fécondité, le taux de reproduction et la survie des adultes d'Aphidius sonchi Marshall (Hym. Aphidiidae, parasite des larves de 2e et 3e stades. A des densités inférieures à 50 pucerons par tige fleurie de Sonchus oleraceus L, par femelle et par jour, la limitation de la ponte due à l'absence d'hôtes a réduit le nombre d'oeufs émis, élevé le taux de superparasitisme et, en conséquence, diminué la fécondité et le taux de reproduction des parasites. Aux densités d'hôtes supérieures, les parasites ont pondu, en moyenne, 220 à 230 oeufs, mais la fécondité réelle et les taux de reproduction ont continué à augmenter avec la densité des pucerons. Par contre, la longévité des parasites n'a pas été affectée par la densité des hôtes, avec une durée moyenne de vie de 4 à 6 jours. L'analyse des données a montré que le taux d'accroissement intrinsèque (r_m) du parasite a changé avec la densité des hôtes, et pourrait atteindre des valeurs supérieures à celles de l'ôte sous des conditions identiques. Les réponses de r_m aux changements de densité des hôtes et au taux sexuel du parasite sont expliquées.Globalement, A. sonchi a présenté une réponse fonctionnelle convexe typique à la densité des hôtes. Cependant, cette réponse a changé au cours de la vie des images et, de plus, les taux de changement ne sont pas logiques à toutes les densités d'hôtes La fréquence de distribution des oeufs n'est généralement pas séparable d'une distribution au hasard, et le nombre d'hôtes parasites peut être prédit d'une façon satisfaisante en utilisant une équation de ponte au hasard.

     

Stylet penetration by the bayberry whitefly, as affected by leaf age in lemon, Citrus limon

Bayberry whitefly (Parabemisia myricae [Kuwana]) crawlers were placed on young and mature lemon leaves and were allowed 7–9 days to settle. Afterwards, the nymphs were fixed and sectioned in situ on the leaves and the area of leaf under each whitefly was examined at 1 000 x for stylet penetration. Both stylets and stylet tracks were readily visible in the sections. The path of penetration was mostly intercellular and the objective appeared to be the phloem. Passage of the stylets through the plant tissue did not cause detectable damage to most cells; however, damaged plant cells occasionally were noticed. Nymphs that had moulted during the 7–9 day settling period reached the phloem significantly more often than those that were still in their first instar. In each of the three replicates, penetration in the mature leaf occurred significantly less often than penetration in the young leaf (4% vs. 72%, p<0.01, ²). Penetration appears to be inhibited in the mature leaf either by the leaf cuticle or by factors detected by the nymphs after very shallow penetration into the leaf. The cuticle of mature leaves was much thicker than the cuticle of young leaves and may have been a barrier to stylet penetration.

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Résumé Des larves de premier stade (avant la fixation) de l'aleurode, Parabemisia myricae (Kuwana), ont été placées, pour qu'elles se fixent, pendánt 2 à 9 jours, sur des feuilles jeunes ou mûres de citronnier. Des morceaux de feuilles avec des larves fixées ont été trempés dans 2% d'agar; ce procédé a permis de maintenir les larves à leurs sites de fixation finale. Des pièces d'agar contenant les morceaux de feuilles avec les larves ont ensuite été trempées dans de la paraffine, et sectionnées en séries de 10 . Il était nécessaire de maintenir une basse température pendant l'opération afin d'obtenir de bonnes sections des feuilles mûres durcies. Les stylets et leurs traces étaient faciles à voir dans les sections teintées aux safranins, et fast green. Presque toutes les traces des stylets étaient intercellulaires et leur destination semblaient être de phloème. En général, la plupart des cellules ne semblaient pas endommagées, les larves qui avaient mué pendant la période de 7 à 9 jours de fixation avaient atteint significantivement plus le phloeme que les larves du premier stade. Dans trois essais, la pénétration des feuilles mûres était significativement moins fréquente que celle des feuilles jeunes (p<0.01, ²).La pénétration dans des feuilles mûres semble être empêchée par la cuticule ou par des facteurs perçus par les nymphes après une pénétration superficielle. La cuticule des feuilles mûres était beaucoup plus épaisse que la cuticule des jeunes feuilles et pourrait donc représenter une barrière à la pénétration des stylets.

     

Food selection by locusts: The rôle of learning in rejection behaviour

The food selection behaviour of male fifth instar nymphs of Locusta migratoria was monitored on the host plant wheat and on the non-host plants Senecio vulgaris, S. jacobaea and Brassica oleracea. The non-hosts were rejected, but the mode of rejection altered with time in a way which suggested associative learning. This hypothesis was tested and the results discussed in relation to classical theories of learning.

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La sélection des aliments chez les criquets: Le rôle de l'apprentissage dans le comportement de rejet

Résumé Des larves mâles de cinquième stade de Locusta migratoria ont été placées dans des cages avec des végétaux qui pouvaient être changées sans troubler les insectes. Le contact avec des pieds de blé entraînait généralement une palpation suivie de morsures et alors d'alimentation continue. Les plantes non-hôtes Senecio vulgaris, S. jacobaea et Brassica oleracea étaient rejetées, d'abord après morsures suivant la palpation, mais ultérieurement par palpation seule. Nous avons vérifié l'hypothèse que la sensation obtenue par palpation était initialement inadéquate pour provoquer le rejet, ce qui était assuré par les morsures ultérieures mais qu'avec l'expérience l'insecte apprenait à lier la sensation de palpation avec celle de morsures et progressivement rejetait après palpation seule. Ainsi les 8 premiers contacts avec des aliments désagréables ont été observés, soit quant S. vulgaris était présenté continuellement, soit quand S. vulgaris était remplacé par une autre plante désagréable après 4 contacts. Une analyse mathématique des résultats révèle une tendance au rejet par palpation seule à travers les contacts successifs, et que cette tendance est interrompue et se restaure quand l'espace végétal est changé. Ces résultats sond discutés dans le contexte des théories de l'apprentissage et on en a conclu que le phénomène observé est le résultat d'un apprentissage associatif.

     

Trypanosoma brucei: analysis of relapsing populations in sensitive and resistant breeds of cattle

The clone DiTat 1.1 of Trypanosoma brucei brucei was injected into four bovids, and clones obtained from successive waves of parasitemia were used to study the expressed variant-specific surface glycoprotein repertoire. Twenty-four clones were obtained which could be classified into 12 different variable antigen types, in addition to the clone injected, using agglutination or immunofluorescence with monospecific antisera. The variable surface glycoproteins of the 25 clones were extracted using the detergent octyl-beta-D-glucopyranoside in the presence of the protease inhibitor, N-cbz-L-phenylalaninechloromethylketone. The molecular weights varied from 52,000 to 69,000 and the pI from 5.0 to 8.8. The virulence of 14 clones representing 13 variable antigen types was ascertained in mice. The mean survival time ranged from 20.5 to 43.0 days. Clones isolated from early peaks of parasitemia in the bovid were the most virulent while clones derived from later peaks were less virulent. It seems that organisms of diminishing virulence appear in bovids, leading to self-cure of the disease. All clones were sensitive to human serum in a blood infectivity inhibition test. Antibody against all virulent clones appeared in 20 cattle (10 Zebus, 10 Baoulés) which had been injected with T. brucei DiTat 1.1. There was no evidence for parasites of high or low virulence being preferentially expressed in resistant or sensitive hosts.     

Sister chromatid differential staining and NOR activity of BrdU-resistant sublines

Summary Two 30 g/ml BrdU-resistant sublines and two 60 g/ml BrdU-resistant sublines are induced from a Chinese hamster cell line Wg3h (HGPRT^–) by one-step and two-step selections, respectively. By inoculating the cells into BrdU-free medium or by adding more BrdU into the culture medium for 26–27 h, it was found that the two BrdU-resistant sublines analysed have very clear sister chromatid differential (SCD) staining patterns. This indicates that some of the nuclear DNA of the BrdU-resistant cells incorporate with BrdU to reach a kinetic balance. Frequencies of sister chromatid exchange (SCE) of the resistant cells are twice to four times as high as those of the Wg3h cells, depending on which BrdU-resistant subline is analysed. The SCE frequencies of the resistant cells also increase with the BrdU concentration in the medium. Analysis of silver-stained nucleolar organizer regions (NORs) indicates that the NOR activity of three out of the four BrdU-resistant sublines is significantly suppressed, i.e., averages of the Ag-NOR number and number of the chromosomes bearing Ag-NORs per cell decrease significantly. The degree of suppression for different BrdU-resistant sublines may be quite different. The suppressed NOR activity of the resistant cells can gradually be restored when the cells are inoculated into BrdU-free medium, but the recovery speed is far lower than that of the Wg3h cells. The suppression of the NOR activity of the BrdU-resistant sublines should be due to BrdU toxicity.     

Evaluation of the Subhuman Primate Pregnancy Test Kit for the detection of early pregnancy in the rhesus monkey (Macaca mulatta)

The performance of The Subhuman Primate Pregnancy Test Kit was evaluated for routine detection of early (days 19-21) pregnancy in the rhesus monkey. Out of 123 confirmed matings, 19 resulted in pregnancy. In the pregnant animals the kit had an accuracy of 73.7%. In the nonpregnant females the accuracy was higher, 88.5%. False positives were encountered in ovariectomized females as well as adult intact males.     

Restriction endonuclease EcoO109 from Escherichia coli H709c with heptanucleotide recognition site 5'-PuG/GNCCPy

A new restriction endonuclease, EcoO109, has been isolated from Escherichia coli H709c by polyethyleneimine (PEI) precipitation, DEAE-cellulose chromatography and heparin agarose chromatography. The yield was high, more than 3000 units/g of wet cells. The EcoO109 endonuclease recognizes and cleaves a nucleotide sequence of (formula: see text), in the presence of 10 mM Mg2+. The enzyme will be useful for structural analysis and molecular cloning of DNA because of the stability, high yield and easy handling of the producer strain.     

用PFU法研究微型生物群集过程中数据的处理

根据MacArthur-Wilson的岛屿区系平衡模型S_t=S_(eq)(1-e~(GT)),可以从野外生态效应试验和室内毒性试验中,提出3个功能参数(S_(eq)、G、t_(90％))进行比较。本文提出两种计算方法:复合梯形法和最小二乘法,后者已在计算机上实现了BASIC计算程序。从数学理论上论证,最小二乘法误差较小,但如果实验布局合理,两种计算方法能得到十分一致的结果。实验模型是否符合理论模型,可以用统计学上的拟合差异度检验法来检验。