Salinity stress proteins in Eurytemora affinis

Seasonal densities of Eurytemora affinis, a calanoid copepod in the Chesapeake Bay, seem to be controlled by temperature and salinity. To examine the role of osmotic stress we analyzed protein synthesis under various conditions of temperature and osmotic stress. Adult females were exposed in groups for 5 hours to different temperature and salinity regimes in the presence of isotope-labelled amino acid. Newly synthesized (stress) proteins could be separated and identified using polyacrylamide gel electrophoresis and autoradiography. The protein profiles occurring in copepods experiencing osmotic shock alone were different from those of control animals. Copepods transferred to lower (2 and 5) and higher (15 and 20) salinities showed differences in the up- and down-regulation of specific proteins. Concurrent heat stress changed these protein patterns. Animals experiencing osmotic and heat shock at the same time exhibited enhanced expression of another set of proteins. Variation in induced proteins occurred among individuals.     

The ecology of the calanoid copepod Eurytemora affinis in salt marsh tide pools

We describe selected aspects of the ecology of the copepod Eurytemora affinis in tide pools of an inland salt marsh near L'Isle-Verte, Québec along the southern shore of the St. Lawrence estuary. E. affinis performed daily horizontal migrations moving from the centers of pools to the banks and into dense algae. Male E. affinis were mainly found in the center of the pools during twilight (21 : 00 hrs) and in dense algae in daylight (12: 00 hrs) whereas most females and copepodites were found near the banks at all three sampling periods. Although these daily movements among sites may have minimized predation by diurnally foraging sticklebacks (Pisces: Gasterosteidae), other explanations for the movements can not be excluded. We also quantified the effects of fish predation upon the population structure of E. affinis. Densities of all stages (nauplius, copepodite, adult) were significantly lower in pools with fish than in pools without fish. Female E. affinis were significantly smaller (mean length) in pools with fish than in pools without fish, indicating that the sticklebacks selectively ate larger females. Male-biased sex ratios were found in both types of pools, which excluded the possibility that biased ratios in this species are caused by selective predation upon the females.     

Long-term changes in the population of Eurytemora affinis (Copepoda,Calanoida) in the Gironde estuary (1978–1992)

Long-term changes in the population of Eurytemora affinis were investigated during 15 years (1978–1992) at a fixed station in the Gironde estuary (South West France). Total numbers, sex-ratio, % ovigerous females, proportion of copepodites and clutch-size were taken into account and their long-term distribution was related to temperature, river flow, salinity, suspended matter and chlorophyll a concentrations after the seasonal effect was removed for all variables.There was a great decrease of the river flow during the period of investigation, due to a general deficit in pluviosity. On the contrary, the general trend for salinity was an increase from 1978 to 1992. The same pattern was observed for chlorophyll, indicating an intrusion of marine phytoplankton. Water temperature increased significantly during the study period, following the general increase in air temperature observed in the area. For the suspended particulate matter (SPM), an increase was observed between 1978 and 1981–1982, then a very sharp decrease occurred from 1984 onwards.Numbers of E. affinis were inversely correlated with temperature and salinity, and positively correlated with the river flow. E. affinis was negatively correlated with chlorophyll concentration because of the covariation with salinity. No clear long-term trend was observed for the sex-ratio. A significant correlation was found between females carrying egg-sacs and SPM concentration, probably due to a decrease of the predation pressure in very turbid waters. The percentages of copepodites tended to decrease with time and were inversely correlated with temperature. Clutch-size significantly decreased during the 15-year period. This trend was mainly explained by temperature and salinity.It is concluded that, in the absence of strong human alteration in the Gironde estuary the long-term distribution of zooplankton can be explained by the natural environmental variability. The dominant factors are the river flow which governs the movement of the populations along the estuary and the temperature which influences the reproductive processes. Any change in climate will have consequences on the pelagic community.     

Some reflections on the structure of the mandibleplate ofEurytemora affinis (Copepoda,Calanoida)

The mandibleplates of several specimen of the copepodEurytemora affinis collected from the Westerschelde and the Gironde estuary were studied. Using the Edge index (ITOH, 1970) this species could be classified as an omnivore. Pictures obtained by light and scanning electron microscopy reveal that the mandible teeth ofE. affinis have a molar-like structure, which likely changes in functional morphology with developmental stage, sex and age of the adults. Based on these observations, the desirability to consider the three dimensional structure of copepod mandibles for feeding mode classification is discussed.Communication no. 540 of the Delta Institute for Hydrobiological Research, Vierstraat 28, 4401 EA Yerseke, The Netherlands.     

Is the copepod egg production in a highly turbid estuary (the Gironde, France) a function of the biochemical composition of seston?

The influence of the biochemical composition of particles originating from surface waters of the Gironde estuary on egg production rates of Eurytemora affinis zooplanktonic population was studied. In the high turbidity zone, suspended particulate matter had a low nutritional quality because the easily available organic fraction represented less than 15% of the overall particulate organic matter. In waters located seaward of the high turbidity zone, a slight increase in nutritional quality was observed. As a result, the sum of easily extractable organic macromolecules represented 15 to 33% of the overall particulate organic matter. The present study suggests that the low egg-production rate of Eurytemora affinis, occurring in the high turbidity zone, results from combined effects of temperature and bad feeding conditions in the area. Low copepod production can be explained by little phytoplankton growth due to light limitation and, therefore, restricted food availability, as well as difficulties in food selection, non-living particle may being dominant.     

Surfactant-producing microorganisms isolated from the gut of a Eucalyptus-feeding sawfly,Perga affinis affinis

Summary Microorganisms which produce surfactants were isolated from the gut of a eucalypt-feeding sawfly. It is possible that surfactants in the guts of some defoliators may be microbiologically-produced.Deceased     

Induction and termination of diapause inOrius predatory bugs

Photoperiodic induction of reproductive diapause at 18°C was investigated in fourOrius [Heteroptera: Anthocoridae] species.Orius insidiosus (Say) displayed a long-day response with a critical photoperiod between L11:D13 and L12:D12. Diapause in this species was terminated rapidly when the temperature and/or the daylength were increased.Orius majusculus (Reuter) also displayed a long-day response. The critical photoperiod fell between L14:D10 and L16:D8. Diapause in this species was not terminated within 14 days when both temperature and daylength were increased. InOrius albidipennis (Reuter) no diapause could be induced at photoperiods varying from L8:D16 to L16:D8. InOrius tristicolor (White) a high proportion of diapause was found at all photoperiods tested. The effect of temperature on photoperiodic induction of diapause was studied inO. insidiosus at L10:D14. Diapause occurred at 18°C, 21°C and 25°C, but not at 30°C. Again, diapause was terminated rapidly after transfer to 25°C/L16:D8. Exposing only the nymphal instars 1–5 to short daylength was not enough to induce diapause in the whole population ofO. majusculus. Orius predatory bugs are used as biocontrol agents against western flower thrips,Frankliniella occidentalis (Pergande) [Thysanoptera: Thripidael, in greenhouses. The consequences of photoperiodic induction of diapause for the success of early season releases ofOrius are discussed.     

Effect of photoperiod on the maternal induction of an egg diapause in the grasshopper Chorthippus bornhalmi

The effect of the photoperiod experienced maternally on the induction of an egg diapause was studied in the grashopper Chorthippus bornhalmi Harz (Acridoidea: Gomphocerinae), which lives on the southern Balkan peninsula. Photoperiods used ranged from L12:D12 to L20:D4 and were kept constant, or were increased or decreased after the imaginal ecdysis in other experiments. Under most conditions in the study, the females laid almost only diapause eggs (not less than 93%). Using constant day-lengths, only at L14:D10 the percentage of non-diapause eggs was somewhat higher (20%). An increase in day-length from moderately long (L14:D10) to long day (L16:D8) was necessary for non-diapause eggs to predominate. However, the percentage of non-diapause eggs decreased with the age of the females, except when the day-length was raised a second time. An increase from short day (L12:D12) to moderately long day (L14:D10) or from long day (L16:D8) to extremely long day (L18:D6) as well as a decrease of day-length, all resulted in the production of diapause eggs. The consequences for the life cycle are discussed.

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Zusammenfassung Der einfluß der Photoperiode auf die maternelle Induktion einer Eidiapause beim Grashüpfer Chorthippus bornhalmi wurde untersucht. Die Art lebt auf der südöstlichen Balkanhalbinsel. Die getesteten Photoperioden reichten von L12:D12 bis L20:D4. In einem Teil der Versuche wurde die Photoperiode während des gesamten Lebens der Grashüpfer konstant gehalten, in anderen nach der Imaginalhäutung erhöht oder erniedrigt. Unter den meisten Bedingungen legten die Grashüpfer-Weibchen fast ausschließlich Diapause-Eier (mehr als 93%). Bei konstanten Tageslängen stieg der Anteil der Subitaneier lediglich nach einer Eiablage bei L14:D10 auf 20%. Eine Erhöhung der Photophase von 14 auf 16 Stunden war notwendig, damit der Anteil an Subitaneier überwog. Aber auch unter diesen Bedingungen nahm ihr Anteil mit dem Alter der Weibchen ab, außer wenn die Photophase noch ein zweites Mal (auf 18 Stunden) verlängert worden war. Eine Zunahme der Tageslänge von Kurztag (L12:D12) auf mäßigen Langtag (L14:D10) oder auch von Langtag (L16:D8) auf extremen Langtag (L18:D6) sowie eine Abnahme der Tageslänge hatten zur Folge, daß die Weibchen fast ausschließlich Diapause-Eier legten. Auf grund der spezialisierten Form der Diapause-Induktion können die Grashüpfer im Freiland nur im Frühjahr Subitaneier legen.

     

Photoperiodic sensitivity and diapause induction during ovarian,embryonic and larval development of the flesh fly,Sarcophaga argyrostoma

Sensitivity to the daily photoperiod, particularly with respect to pupal diapause induction, was studied during ovarian, embryonic, and larval development of the flesh flySarcophaga argyrostoma. Large flies were shown to have a greater number of primary follicles in their ovaries and to be capable of limited ovarian maturation in the absence of exogenous protein (autogeny). Such ovarian development occurred independently of photoperiod. However, long days experienced during embryogenesis caused more rapid development, and earlier larviposition, than short days. Short days during embryonic and subsequent larval development also induced pupal diapause, whereas long days led to continuous or non-diapause development of the pupae. Pupal diapause could not be induced by photoperiods during the vitellogenic phase of ovarian development. InSarcophaga argyrostoma, a maternal effect preventing pupal diapause among the progeny of files with a diapause history was not observed.     

The ultrastructure of the eye of the mosquitofishGambusia affinis

Summary The ultrastructure of the tissue components of the eye ofGambusia affinis, excluding the sensory cells, is described. The cornea consists of two different sections of collagenous layers of different density. The choroid includes an argentea composed of- and-melanophores, lipopterinophores and a choriocapillaris associated with the rete mirabile of the choroid body. Bruch's membrane, underlying the retinal pigment layer, can develop complex associations with fibroblasts delimiting the choriocapillaris. The outer section (stroma) of the iris includes several cell types that are not found in the inner or vitread section. In adultGambusia the lens capsule is well developed, but in twoweek-oldSarotherodon larvae the lens epithelium is covered only by a glycocalyx.     

Control of diapause in codling moth larvae

Diapause in a New Zealand strain of codling moth (Cydia pomonella Linnaeus [Lepidoptera: Olethreutidae]) was induced in larvae by photoperiods of 15 h or less. Once diapause had been initiated, it could not be terminated by any combination of conditions tested for at least 20 days after cocooning. In diapausing larvae a low rate of pupation occurred at 25 °C under a long day (18 h) photoperiod. A high rate of pupation was achieved under a long day regime when larvae were decocooned, and provided with apple as nourishment. Diapause could be terminated predictably in 94–100% of larvae by 1) conditioning at 15 °C and constant darkness for periods of 40–100 days, then 2) chilling at 2±2 °C and constant darkness for 20–50 days followed by 3) any post-chill condition periods at 25 °C, 18 h photoperiod. Complete diapause termination was achieved when 100 days conditioning was followed by 30 days or 50 days post-chill period. Under these conditions, 76% termination occurred in the post-chill period after 10 days, and 93% after 25 days.To terminate diapause in codling moth larvae, we recommend that a 100 days conditioning followed by 30 days chilling and 50 days post chilling periods be used.     

Termination of diapause in the Cinnabar moth (Tyria jacobaeae) by chilling

The effect of chilling time on the termination of pupal diapause was studied in the Cinnabar moth, Tyria jacobaeae L., Lepidoptera: Arctiidae. Chilling followed by a temperature of 20°C was effective in terminating diapause. Chilling for 120 days immediately after pupation yielded the highest percentage emergence and resulted in the shortest total pupal development time (162 days). A more natural treatment, a warm period before cold treatment, did not result in a higher percentage of emergence or in a shorter total pupal development time.During hibernation this species experiences two successive processes: diapause development during the first three to four months of cold, followed by a period of quiescence. Application of high temperatures in this latter period immediately starts morphogenesis. The impact of a relative humidity of 70% during the pupal stage for complete wing unfolding is demonstrated. The underlying mechanism of diapause is discussed.

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Résumé L'effet sur la levée de la diapause de différents régimes de température infligés aux chrysalides est étudié chez Tyria jacobaeae L. (Lepidoptera: Arctiidae). Une mise au froid suivie de hautes températures lève efficacement la diapause. Un refroidissement pendant 120 jours immédiatement après la nymphose entraîne le plus grand pourcentage d'éclosions et la durée totale du développement des chrysalides la plus courte (162.3 j). Un traitement plus naturel, une période chaude avant la période froide, n'entraîne pas un plus grand pourcentage d'éclosions ni une durée plus courte de développement.Il est proposé que durant l'hibernation cette espèce passe par deux processus successifs: un diapause durant les premiers trois ou quatre mois de froid, suivie d'une période de quiescence. Le transfert à des hautes températures pendant cette derpériode déclanche immédiatement la morphogénèse. L'influence d'un taux élevé d'humidité relative durant le stade chrysalide sur le déploiement complet des ailes est démontré. Le méchanisme de diapause sous-jacent est discuté.

     

Establishment of in vitro plants, cell and tissue cultures from Oldenlandia affinis for the production of cyclic peptides

In vitro cultured plants from Oldenlandia affinis were established from seeds and grown on a hormone-free medium. In vitro plants produced the cyclic peptide kalata B1 in concentrations of 0.67 mg g⁻¹ dry weight after growth of 30 days. This was approximately 50% of the concentration analysed in green house plants (shoot tips), where different concentrations have been determined in leaves (1.82 mg g⁻¹), shoot tips (1.36 mg g⁻¹), stems (0.36 mg g⁻¹), and in flowers (0.16 mg g⁻¹). Callus and cell suspension cultures could be initiated from aseptic root, stem and leaf explants of O. affinis seedlings and plants. Different light intensities were shown to affect culture growth as well as chlorophyll synthesis. The friable callus was then used for the establishment of a cell suspension culture. Fresh and dry weight measurements showed that growth was optimal on MS medium supplemented with 0.4 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-d). Leaf suspensions cultured on this medium showed a 4-fold increase of biomass by the first week of incubation. No quantifiable amounts of kalata B1 were produced under these conditions. Morphological differentiation seems to be essential for cyclic peptide production. Therefore, several undifferentiated as well as organised cell lines of O. affinis have been developed. These cell lines will constitute a worthwhile starting point for the optimisation of kalata B1 synthesis in liquid media to the objective of producing cyclic peptides under controlled and defined conditions in bioreactors.     

Induction of reproductive diapause in Bruchidius atrolineatus during the dry season in a Sahelian zone

Bruchidius atrolineatus (Pic) (Bruchidae: Coleoptera) is a bruchid beetle infesting Vigna unguiculata (Walp). During the dry season, adults are in reproductive diapause. Studies of adult populations in a traditional store containing contaminated pods of V. unguiculata show that the reproductive diapause is induced during the first larval instars. The temperature decrease from November to January probably is the main climatic factor inducing this diapause. The percentage of diapausing adults depends on the duration of the larval development at low temperatures. Experiments in climatic boxes confirm these results. At 12 h 25 °T-12 h 20° C 11–13 LD, a high proportion of the adults emerging from the V. unguiculata seeds are in reproductive diapause and the duration of their growth is variable. L1 L2 L3 larvae probably are sensitive stages to the thermal variations and the processes allowing emergence of diapausing adults probably are induced during these larval instars.

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Résumé Bruchidius atrolineatus (Pic) est un Coléoptère Bruchidae se développant en zone sahélienne aux dépens d'une légumineuse Vigna unguiculata. Pendant la saison sèche, les adultes sont en diapause reproductrice. Un suivi des populations d'adultes a été réalisé dans un grenier traditionnel en terre séchée contenant des gousses récoltées dans la nature et portant des ufs de B. atrolineatus. Il a permis de constater que la diapause reproductrice était induite durant les premières phases du développement larvaire. La décroissance des températures de Novembre à Janvier avec des minima de 15 ° C et des maxima de 25 °C est probablement le principal facteur permettant l'induction de cette diapause. L'introduction dans le grenier de gousses portant des ufs venant d'être émis à différentes périodes de la saison sèche montre que le pourcentage d'adultes diapausants dépend de la durée du développement larvaire aux températures basses. Des expériences réalisées dans des étuves programmées confirment ces résultats. A 12 h 25 °T/12 h 20 °C 11–13 LD, un taux élevé d'adultes émergeant des graines est en diapause reproductrice et la durée de leur développement post-embryonnaire est très variable. En conditions plus chaudes 12 h 40 °C/12 h 25 °C 11–13 LD, le taux d'adultes diapausants est plus réduit et la durée du développement plus courte. L'analyse de la croissance larvaire montre que les premiers stades larvaires sont probablement des stades sensibles aux variations thermiques. Les modifications physiologiques permettant l'émergence d'adultes diapausants ou sexuellement actifs seraient induites durant ces stades.

     

L'induction maternelle de la diapause larvaire chezDinarmus acutus [Hym.: Pteromalidae]

Résumé Dans la région béarnaise, la récolte des gousses m?res deLathyrus sylvestris et deLathyrus latifolius, avant la déhiscence, puis la mise en éclosoir de leurs graines, permet l'obtention des adultes de la brucheBruchus affinis et de son ectoparasite larvaireDinarmus acutus. Ces derniers sortent en petits nombres à l'automne de la même année puis de fa?on massive au printemps de l'année suivante, après une diapause au dernier stade larvaire. Un élevage en continu, de nombreuses générations successives, en conditions semi naturelles de température et de photopériode, a montré la présence simultanée d'individus diapausants et non diapausants tout au long de l'hiver, et l'absence totale de diapausants dès le mois de mai. Les larves diapausantes produisent une substance orangée qui marque l'arrêt de développement. Compte tenu de la biologie de la bruche et de son parasite larvaire: développement larvaire à l'intérieur des graines et vie des adultes à l'extérieur, nous avons choisi d'étudier l'influence des conditions thermiques sur les larves et de la photopériode sur les adultes du parasite. Les résultats obtenus montrent que les températures basses n'induisent pas d'arrêt de développement. Par contre, une photopériode jours courts, appliquée aux adultes, provoque l'entrée en diapause d'un fort pourcentage des larves de la descendance. Les résultats présentent une certaine variabilité selon la descendance et le rang de la ponte. Le détail des résultats est examiné dans le cadre de l'induction maternelle de la diapause larvaire et de l'hypothèse d'une horloge chimique à l'origine du marquage des diapausants (substance orangée) et de la variabilité enregistrée.      

Development of 19 polymorphic microsatellite loci for the intermediate horseshoe bat, Rhinolophus affinis (Rhinolophidae, Chiroptera)

The intermediate horseshoe bat (Rhinolophus affinis) is a widespread species in Southeast Asia. We developed 19 novel microsatellite loci from an enriched genomic library of the bat, and tested their polymorphism using a single population from Guangdong province, southern China. The number of alleles ranged from 3 to 15 per locus with the expected and observed heterozygosity ranging from 0.397 to 0.920 and 0.280–0.926, respectively. Three markers significantly deviated from the Hardy-Weinberg expectations after Bonferroni correction and no linkage disequilibrium was detected in any of loci. These microsatellite loci will be useful in studying the phylogeography of this species.     

Molecular mechanism of diapause in insect (I) --Expression of diapause hormone gene and incubation temperature in embryonic stage of Bombyx mori

The embryonic diapause of the silkworm, Bombyx mori, is induced by the diapause hormone (DH) which is secreted from the suboesophageal ganglion of pupae. The diapause nature of bivoltine strains uses environmental stimuli as the initial signal to determine the diapause nature. The experiments showed that DH gene expression is a direct response to the environmental stimulus, such as high incubation temperature. The cDNA from the embryonic stage was cloned and sequence analysis showed the cDNA encoding DH. Expression patterns of the DH gene in embryonic stage are different at incubation temperatures 15℃ and 25℃, suggesting that the incubation temperature as an environmental signal is kept within the body to control the DH gene expression at the pupal stage, so that the embryonic diapause of next generation can be determined.     

Aspects of habitat selection in the mosquitofish gambusia affinis

Laboratory experiments were performed to determine behavioral preferences of Gambusia affinis for habitat variables in partitioned aquaria. The fish preferred calm water, dark-colored substrates and subsurface vegetation providing lateral concealment. Floating cover, which obstructed access to the surface, was avoided. Lack of preference for real over plastic plant cover indicates that visual rather than chemical cues are involved. These laboratory preferences correspond to the reported microhabitat distribution of G. affinis in nature and indicate the presence of an active habitat preference, as opposed to passive habitat correlation, in this species. Species-specific habitat preferences, which may be narrower where a species occurs sympatrically with its congeners, probably form part of a reproductive isolating mechanism.