Emergence rhythms and protandry in relation to daily patterns of locomotor activity inTrichogramma species

Summary In insect parasitoids, the mating structure of the population depends on the emergence pattern of adults and on their behaviour after emergence. In order to study some aspects of these characteristics in four species of egg parasitoid of the genusTrichogramma, laboratory experiments using automatic video analysis are performed. Adult emergence is concentrated in the morning, with an earlier time for males. The variability of the time of emergence is under genetic control. The relationships between the emergence pattern and the locomotor activity are considered by measuring individually: first, the delay between the emergence and the first locomotion which is significantly greater in females than in males and, second, the activity of newly emerged males, which is maximum at the time of the emergence of females. The locomotor activity is influenced by mating; mated females are much more active than virgin ones, mated males are less active than virgin ones. The adaptive values of the rhythmicity of emergence and of protandry are discussed together with their relationship with the patterns of locomotor activity. The results are also considered with respect to mating strategies in insect parasitoids; they actually support the hypothesis of mating occurring principally among individuals native to the same host patch inTrichogramma species.     

Evolution of circadian rhythms in Drosophila melanogaster populations reared in constant light and dark regimes for over 330 generations

Organisms are believed to have evolved circadian clocks as adaptations to deal with cyclic environmental changes, and therefore it has been hypothesized that evolution in constant environments would lead to regression of such clocks. However, previous studies have yielded mixed results, and evolution of circadian clocks under constant conditions has remained an unsettled topic of debate in circadian biology. In continuation of our previous studies, which reported persistence of circadian rhythms in Drosophila melanogaster populations evolving under constant light, here we intended to examine whether circadian clocks and the associated properties evolve differently under constant light and constant darkness. In this regard, we assayed activity-rest, adult emergence and oviposition rhythms of D. melanogaster populations which have been maintained for over 19 years (~330 generations) under three different light regimes – constant light (LL), light–dark cycles of 12:12 h (LD) and constant darkness (DD). We observed that while circadian rhythms in all the three behaviors persist in both LL and DD stocks with no differences in circadian period, they differed in certain aspects of the entrained rhythms when compared to controls reared in rhythmic environment (LD). Interestingly, we also observed that DD stocks have evolved significantly higher robustness or power of free-running activity-rest and adult emergence rhythms compared to LL stocks. Thus, our study, in addition to corroborating previous results of circadian clock evolution in constant light, also highlights that, contrary to the expected regression of circadian clocks, rearing in constant darkness leads to the evolution of more robust circadian clocks which may be attributed to an intrinsic adaptive advantage of circadian clocks and/or pleiotropic functions of clock genes in other traits.     

The statistical analysis of tidal rhythms: Tests of the relative effectiveness of five methods using model simulations and actual data

Endogenous rhythms in intertidal organisms are often very complex and imprecise. Thus, confusing results are sometimes obtained when applying various interpretive analytical techniques. In an attempt to resolve this problem, ten different models representing typical organismic tidal‐rhythm displays were created and examined with five different inferential statistical techniques. The exercise was designed to test the relative effectiveness of these techniques in detecting the presence of known cycles in the models, and estimating their period lengths. The same comparison was then repeated on sets of animal‐derived data.

All of the five methods had their merits, but, depending on the model being examined, the results from the various methods were not identical. Three of the techniques produce harmonics, making data that contain multiple periods especially difficult to decipher. Often both tidal and circadian periods are displayed by shore dwellers; all five methods were able to find these two periods. But when the difference in circa period length was close, only one technique (array analysis) could make a distinction. This technique was also the only one able to handle data in which the period was not constant. Interestingly, this simplest of methods is probably the best all‐round method of discovery. Many more subtle, but important, differences were also noted, and it is recommended that more than one method always be used to ensure accuracy.     

Effects of a quantitative trait locus for increased muscularity on carcass traits measured by subjective conformation and fat class scores and video image analysis in crossbred lambs

A quantitative trait locus (QTL) for increased loin muscularity (TM-QTL) has previously been identified in purebred Texel sheep. Crossbred lambs born out of Mule ewes mated to heterozygous Texel sires for the TM-QTL were evaluated for a range of carcass traits. Lambs were genotyped and classified as carriers (n = 62) of a single copy of the TM-QTL and non-carriers (n = 49). In this study, the effects of the TM-QTL on carcass attributes were investigated using subjective classification scores for conformation and fatness, and measurements from a video image analysis (VIA) system. In addition, refined prediction equations to estimate weights of primal joints (leg, chump, loin, breast and shoulder) were obtained by calibrating the VIA system against computer tomography (CT) measurements in the loin region. The new refined prediction models increased the accuracy of prediction of all primal cuts on an average of 16% compared to previously derived standard VIA prediction equations. The coefficient of determination (R2) of the VIA system to predict in vivo CT measurements ranged from 0.39 to 0.72 for measurements of Musculus longissimus lumborum (MLL) area, width and depth, lumbar spine length, loin muscle volume and loin muscularity index. Using VIA estimates of CT-measured loin muscle traits, a significant increase in depth (+2.7%) of the MLL was found to be associated with the TM-QTL. Conformation and fatness scores and the shape of the carcass measured as individual lengths, widths and areas by VIA were not significantly influenced by the TM-QTL. Primal meat yields estimated using both standard and refined VIA prediction equations were not significantly affected by the TM-QTL. However, carcass 'compactness' was found to have significantly increased in carrier lambs. The weight of the dissected MLL estimated using VIA information was greater (+2.6%) for carriers compared to non-carriers. To conclude, neither the current industry carcass evaluation system for conformation and fatness nor the standard VIA system is able to identify the effect of the TM-QTL in the loin region in the moment. However, the calibration of the VIA system against CT measurements resulted in improved VIA prediction equations for primal meat yields and also showed moderate potential to estimate loin muscle traits measured by CT and to detect, partially, the effect of the TM-QTL on these traits.     

Quantitative comparison of taxa and taxon concepts in the diatom genus Fragilariopsis: a case study on using slide scanning,multiexpert image annotation,and image analysis in taxonomy1

Semiautomated methods for microscopic image acquisition, image analysis, and taxonomic identification have repeatedly received attention in diatom analysis. Less well studied is the question whether and how such methods might prove useful for clarifying the delimitation of species that are difficult to separate for human taxonomists. To try to answer this question, three very similar Fragilariopsis species endemic to the Southern Ocean were targeted in this study: F. obliquecostata, F. ritscheri, and F. sublinearis. A set of 501 extended focus depth specimen images were obtained using a standardized, semiautomated microscopic procedure. Twelve diatomists independently identified these specimen images in order to reconcile taxonomic opinions and agree upon a taxonomic gold standard. Using image analyses, we then extracted morphometric features representing taxonomic characters of the target taxa. The discriminating ability of individual morphometric features was tested visually and statistically, and multivariate classification experiments were performed to test the agreement of the quantitatively defined taxa assignments with expert consensus opinion. Beyond an updated differential diagnosis of the studied taxa, our study also shows that automated imaging and image analysis procedures for diatoms are coming close to reaching a broad applicability for routine use.     

Development of image analysis techniques as a tool to detect and quantify morphological changes in anaerobic sludge: I. Application to a granulation process

Image analysis techniques were developed and applied to quantify the process of anaerobic granulation in an expanded granular sludge blanket reactor (EGSB) fed with a synthetic substrate based on glucose [60-30% COD (chemical oxygen demand)] and volatile fatty acids (40-70% COD) over 376 days. In a first operation period that lasted 177 days, the aggregation of dispersed sludge was quantitatively monitored through the recognition and quantification of aggregates and filaments. A parameter defined as the ratio between the filaments' length and the aggregates projected area (LfA) has proven to be sensitive to detect changes in the aggregation status of the anaerobic sludge. The aggregation time-defined as the moment when a balance between filaments' length and aggregates' size was established-was recognized through the LfA. The percentage of projected area of aggregates within three size ranges (0.01-0.1 mm, 0.1-1 mm, and >1 mm, equivalent diameter) reflected the granular size spectrum during the aggregation process. When sudden increases on the upflow velocity and on the organic loading rate were applied to the previously formed granules, the developed image analysis techniques revealed to be good indicators of granular sludge stability, since they were sensitive to detected filaments release, fragmentation, and erosion that usually leads to washout. The specific methanogenic activities in the presence of acetate, propionate, butyrate, and H(2)/CO(2) increased along the operation, particularly relevant was the sudden increase in the specific hydrogenophilic activity, immediately after the moment recognized as aggregation time.     

Development of image analysis techniques as a tool to detect and quantify morphological changes in anaerobic sludge: II. Application to a granule deterioration process triggered by contact with oleic acid

Image analysis techniques are applied to monitor the morphological changes in granular sludge present in an expanded granular sludge blanket (EGSB) reactor fed with oleic acid. Deterioration of granular sludge was monitored along the trial period by measuring the percentage of aggregates smaller than 1 mm (in terms of Feret diameter) either in terms of projected area or in terms of number of aggregates. A good correlation was obtained between these values and the percentage of aggregates smaller than 1 mm were physically sorted and quantified by the volatile suspended solid content. The ratio of total filaments length to cross-sectional area of aggregates defined as LfA, was applied to quantify the dispersion level of the granular sludge, which increased until day 141 and remained almost invariant afterwards. LfA was sensitive to the sludge deterioration process and was able to indicate, with the anticipation of about 1 month, the most significant biomass washout episode that occurred in the trial period. A mechanism of filaments' release, detachment and selective washout was proposed to explain the action of LfA from this viewpoint. The equivalent diameter of the bottom aggregates larger than 1 mm increased with the increase on the amount of long chain fatty acids associated with the biomass by mechanisms of adsorption, precipitation, or entrapment. After a threshold value of about 200 mg COD-LCFA gVSS (COD = chemical oxygen demand; LCFA = long chain fatty acids; VSS = volatile suspended solids), a migration of granular sludge from the bottom to a top-floating layer was evident.