Aerobic Metabolism Alterations as an Evidence of Underlying Deltamethrin Resistance Mechanisms in Triatoma infestans (Hemiptera: Reduviidae).

Triatoma infestans (Klug, 1834), the main vector of Chagas disease in Latin America, is regularly controlled by spraying the pyrethroid deltamethrin, to which some populations have developed resistance. The three main mechanisms of resistance are 1) metabolic resistance by overexpression or increased activity of detoxifying enzymes, 2) target site mutations, and 3) cuticle thickening/modification. We use open-flow respirometry to measure real-time H2O loss rate (V˙H2O) and CO2 production rate (V˙CO2), on nymphs from susceptible and resistant populations before and after exposure to the insecticide to understand the underlying mechanisms of resistance in live insects. Lack of differences in V˙H2O between populations suggested that cuticular thickness/composition is not acting as a relevant resistance mechanism. Similarly, there was no difference in resting V˙CO2, suggesting a trade-off between resistance mechanisms and other physiological processes. The increment in V˙CO2 after application of deltamethrin was similar in both populations, which suggested that while enhanced enzymatic detoxification may play a role in resistance expression in this population, the main mechanism involved should be a passive one such as target site mutations. Open-flow respirometry provided useful evidence for evaluating the mechanisms involved in deltamethrin resistance. Using this technique could improve efficiency of scientific research in the area of insecticide resistance management, leading to a faster decision making and hence improved control results.

Genetic variation for tolerance to high temperatures in a population of Drosophila melanogaster.

The range of thermal tolerance is one of the main factors influencing the geographic distribution of species. Climate change projections predict increases in average and extreme temperatures over the coming decades; hence, the ability of living beings to resist these changes will depend on physiological and adaptive responses. On an evolutionary scale, changes will occur as the result of selective pressures on individual heritable differences. In this work, we studied the genetic basis of tolerance to high temperatures in the fly Drosophila melanogaster and whether this species presents sufficient genetic variability to allow expansion of its upper thermo-tolerance limit. To do so, we used adult flies derived from a natural population belonging to the Drosophila Genetic Reference Panel, for which genomic sequencing data are available. We characterized the phenotypic variation of the upper thermal limit in 34 lines by measuring knockdown temperature (i.e., critical thermal maximum [CTmax]) by exposing flies to a ramp of increasing temperature (0.25°C/min). Fourteen percent of the variation in CTmax is explained by the genetic variation across lines, without a significant sexual dimorphism. Through a genomewide association study, 12 single nucleotide polymorphisms associated with the CTmax were identified. In most of these SNPs, the less frequent allele increased the upper thermal limit suggesting that this population harbors raw genetic variation capable of expanding its heat tolerance. This potential upper thermal tolerance increase has implications under the global warming scenario. Past climatic records show a very low incidence of days above CTmax (10 days over 25 years); however, future climate scenarios predict 243 days with extreme high temperature above CTmax from 2045 to 2070. Thus, in the context of the future climate warming, rising temperatures might drive the evolution of heat tolerance in this population by increasing the frequency of the alleles associated with higher CTmax.

The costs of living in a thermal fluctuating environment for the tropical haematophagous bug, Rhodnius prolixus.

Environmental temperature is an abiotic factor with great influence on biological processes of living beings. Jensen's inequality states that for non-lineal processes, such as most biological phenomena, the effects of thermal fluctuations cannot be predicted from mean constant temperatures. We studied the effect of daily temperature fluctuation (DTF) on Rhodnius prolixus, a model organism in insect physiology, and an important vector of Chagas disease. We measured development time from egg to adult, fecundity, fertility, body mass reduction rate (indirect measurement of nutrient consumption rates) and survival after a single blood meal. Insects were reared at constant temperature (24 °C), or with a DTF (17-32 °C; mean = 24 °C). Taking into account Jensen's inequality as well as the species tropical distribution, we predict that living in a variable thermal environment will have higher costs than inhabiting a stable one. Development time and fertility were not affected by DTF. However, fecundity was lower in females reared at DTF than at constant temperature, and males had higher body mass reduction rate and lower survival in the DTF regime, suggesting higher costs associated to fluctuating thermal environments. At a population and epidemiological level, higher energetic costs would imply an increase in nutrient consumption rate, biting frequency, and, consequently increasing disease transmission from infected insects. On the contrary, lower fecundity could be associated with a decrease in population growth. This knowledge will not only provide basic information to the field of insect ecophysiology, but also could be a useful background to develop population and disease transmission models.

Enhanced fertility and chill tolerance after cold-induced reproductive arrest in females of temperate species of the Drosophila buzzatii complex.

Long-term exposure to low temperatures during adult maturation might decrease fertility after cold recovery as a consequence of carry-over effects on reproductive tissues. This pattern should be more pronounced in tropical than in temperate species as protective mechanisms against chilling injuries are expected to be more effective in the latter. We initially determined the lower thermal thresholds to induce ovarian maturation in four closely related Drosophila species, two inhabiting temperate regions and the other two tropical areas of South America. As expected, only temperate species regularly experience cold-inducing conditions for reproductive arrest during winter in their natural environment. Subsequently, we exposed reproductively arrested and mature females to cold-inducing conditions for reproductive arrest over a long period. Following cold exposure, tropical species exhibited a dramatic fertility decline, irrespective of reproductive status. In contrast, not only were temperate females fecund and fertile but also fertility was superior in females that underwent cold-induced reproductive arrest, suggesting that it might act as a protecting mechanism ensuring fertility after cold recovery. Based on these findings, we decided to evaluate the extent to which reproductive status affects cold tolerance and energy metabolism at low temperature. We found a lower metabolic rate and a higher cold tolerance in reproductively arrested females, although only temperate species attained high levels of chill tolerance. These findings highlight the role of cold-induced reproductive arrest as part of an integrated mechanism of cold adaptation that could potentially contribute to the spread of temperate species into higher latitudes or altitudes.

Aggregated oviposition in Rhodnius prolixus, sensory cues and physiological consequences.

Females of the haematophagous bug Rhodnius prolixus attach their eggs in clusters on substrates related to their hosts, such as nests or avian feathers. Because the hosts are an enormous food resource as well as potential predators, the choice of the site and pattern of oviposition could have an important adaptive value. Here we investigated proximate and a potential ultimate cause of this aggregated pattern of laid eggs. First, we studied proximal causes by analyzing the use of chemical or physical cues associated with aggregated oviposition in R. prolixus. For all terrestrial organisms there is a trade-off between exchange of respiratory gases and water loss. Particularly, insect eggs are highly susceptible to this trade-off because they do not obtain water from the environment, hence our second objective is to study the possible mechanisms involved in dehydration resistance in this species. Therefore we examined the dynamics of change in CO2 release rate (MCO2), and water loss rate (MH2O) in relation to embryo development as energetic demands increase, and tested the energetic or hygric efficiency hypothesis as a potential ultimate cause of aggregated oviposition. This hypothesis states that grouped eggs consume less energy or lose less water than equal numbers of isolated eggs, the latter being more susceptible to dehydration. Results indicated the use of physical external cues such as dummy eggs or edges of the oviposition substrates, but we did not find any chemical cues associated with the aggregated pattern of oviposition. There are no energetic or hygric benefits associated with egg's aggregated pattern. However, when we analyzed the MCO2 and MH2O change in relation to embryo development, we found a fairly constant and low MH2O albeit a clear increase in MCO2, suggesting a tightly control of egg's desiccation tolerance. This high resistance to desiccation coupled with a temporal strategy of hatching allows R. prolixus embryos to successfully develop and hatch under harsh environmental conditions.

Metabolism and water loss rate of the haematophagous insect Rhodnius prolixus: effect of starvation and temperature.

Haematophagous insects suffer big changes in water needs under different levels of starvation. Rhodnius prolixus is the most important haematophagous vector of Chagas disease in the north of South America and a model organism in insect physiology. Although there have been some studies on patterns of gas exchange and metabolic rates, there is little information regarding water loss in R. prolixus. We investigated whether there is any modulation of water loss and metabolic rate under different requirements for saving water. We measured simultaneously CO2 production, water emission and activity in individual insects in real time by open-flow respirometry at different temperatures (15, 25 and 35°C) and post-feeding days (0, 5, 13 and 29). We found: (1) a clear drop in metabolic rate between 5 and 13 days after feeding that cannot be explained by activity and (2) a decrease in water loss rate with increasing starvation level, by a decrease in cuticular water loss during the first 5 days after feeding and a drop in the respiratory component thereafter. We calculated the surface area of the insects and estimated cuticular permeability. In addition, we analysed the pattern of gas exchange; the change from a cyclic to a continuous pattern was affected by temperature and activity, but it was not affected by the level of starvation. Modulation of metabolic and water loss rates with temperature and starvation could help R. prolixus to be more flexible in tolerating different periods of starvation, which is adaptive in a changing environment with the uncertainty of finding a suitable host.