Behavioural circadian rhythms measured in real-time by automatic image analysis: applications in parasitoid insects

Abstract. A video analysis system was developed to study behavioural circadian rhythms in insects. Technical innovations were performed in image analysis and in the use of robotics. In image analysis, both the automatic detection of the insect and the calculation of several variables (linear and angular speed, locomotor activity, etc.) are carried out in real-time, which saves time and allows long-term studies without any intervention of the operator. The displacement of the camera by robotics allows the simultaneous study of many individuals (i.e. sixty individuals for the study of locomotor activity of adult insects, more than 400 individuals for the study of the emergence of adult parasitoids from their host). Four applications of this new device are presented. They deal with circadian rhythms in parasitoid insects: locomotor activity and time variation in locomotory parameters, daily distribution of the emergence and onset of the locomotor activity in adults. The adaptability of the system to several experimental materials and situations is discussed and the interest of the study of behavioural rhythms in the approach of host-parasitoid associations is emphasized.     

Aedes polynesiensis in the Society Islands: environmental correlates of isoenzyme differentiation

Abstract Isoenzyme genetic differentiation of Aedes polynesiensis mosquitoes in Raiatea island, French Polynesia, was evaluated by two models of population structure based on seven gene-enzyme systems: Ak, Est, Got, Gpi, Hk, Mdh and Pgm. The ecological model tested whether genetic differentiation is congruent with habitat differences. The isolation model evaluated whether genetic differentiation is proportional to geographical distribution. The ecological model found no significant differentiation between populations of Ae.polynesiensis from beach and forest ecotopes, whereas the isolation model was consistent with the data. However, incipient speciation is opposed by the source-sink system of population dynamics in such small neighbouring islands, where Ae.polynensiensis extinction is readily followed by reinvasion causing considerable gene flow between island populations.     

Oral susceptibility of Aedes albopictus to dengue type 2 virus: a study of infection kinetics, using the polymerase chain reaction for viral detection

Female Aedes albopictus mosquitoes, aged 1 week, were infected with DEN-2 dengue virus. The kinetics of infection in mosquito brain and mesenteron were monitored using DNA probes with polymerase chain reaction (PCR) amplification of target DNA sequences coding for DEN-2 virus envelope protein, compared with the standard immunofluorescence assay technique (IFA). Rates of virus detection in the mesenteron of orally infected mosquitoes rose to 38% by day 4 post-inoculation, then declined until day 8, followed by irregular peaks around days 11-14 and subsequently. In mosquito head squashes, virus was detected from day 4 onwards, reaching 38% positive by day 18. Salivary glands of all the same females were found to be positive for virus by day 8 onwards. Parenterally infected Ae.albopictus females were all positive for DEN-2 in the brain and salivary glands 8 days post-inoculation. In every case, results obtained with the PCR matched those from the IFA. Our DNA probe with PCR procedure can therefore be utilized as a sensitive and reliable method for studies of DEN-2 vectors.     

Heat Stress-induced Abortion of Buds and Flowers in Pea: Is Sensitivity Linked to Organ Age or to Relations between Reproductive Organs?

A short period of heat stress can cause a significant increasein abortion frequency of floral buds and flowers in pea, buta large variability in sensitivity exists among reproductiveorgans (ROs) within a plant or between plants. We have analysedspatial distribution of abortion frequency in plants subjectedor not subjected to heat stress in: (1) four controlled environmentexperiments in which apex temperature was increased to 31 °Cfor 6 h per day for 4 d (moderate stress); (2) one field experimentwith a similar level of stress; and (3) one experiment with2 d with 33/30 °C day/night (severe stress) in additionto treatment (1). Influence of neighbouring organs on sensitivityto heat stress was analysed by partial flower removal. Severeheat stress caused rapid interruption of RO development followedby abscission while, in moderate stress, at leat 4 d elapsedbetween cessation of stress and first symptoms of abortion.In both moderate stress and control treatments, abortion frequencyfollowed a consistent pattern along the stem with no abortionon the lowest reproductive phytomeres, a temperature-dependentabortion frequency on intermediate phytomeres and 100% abortionon apical phytomeres. Regardless of temperature, this patternwas shifted by four positions if ROs were removed on the lowestfour reproductive phytomeres, and no abortion was observed ifyoung pods were sequentially removed. Moderate heat stress increasedthe effect of developing ROs on abortion frequency of youngerROs located above them. We suggest that the effect of mild stressshould be viewed as the acceleration of a programme linked tothe normal termination of phytomere production during the plantcycle, rather than as an abrupt event linked to stress. Pisum sativum; heat stress; abortion; abscission; age; organ-to-organ relations