DNA repair is efficient in irradiated M phase zygotes.

In somatic cells, DNA repair is attenuated during mitosis to prevent the formation of anaphase bridges and facilitate the proper segregation of sister chromatids. Irradiation-induced γH2AX foci persist for hours in M phase somatic cells. However, we observed that anaphase bridges formed in a significant fraction of mouse zygotes irradiated during mitosis. Additionally, γH2AX signals in M phase zygotes peaked 30 min after irradiation and subsequently reduced with a half-life within 1-2 h. These results suggest that the DNA repair system may operate efficiently in M phase zygotes following irradiation, leading to the frequent formation of anaphase bridges. The absence of H2AX promoted the successful segregation of sister chromatids and enhanced the development of embryos to the blastocyst stage. The DNA repair system may be differentially regulated during the M phase of the first cell cycle to ensure the immediate elimination of damaged zygotes, thereby efficiently preventing transmission of mutations to subsequent generations.

Effects of monochromatic green light stimulation during embryogenesis on hatching and posthatch performance of four strains of layer breeder.

Providing green light during incubation has been shown to accelerate the embryo development and shorten the hatching time in broilers. Few studies have concentrated on the exact effects on layer breeders in the aspects of hatching and posthatch performance. In this study, 4 strains of layer breeder eggs, namely White Leghorn, Rhode Island Red, Columbia Rock, and Barred Rock were used to assess the effects of monochromatic green light during embryogenesis on hatching performance, chick quality, and pubertal growth. Each strain of 600 eggs was incubated under photoperiods of either 12 h of light and 12 h of darkness (12L:12D, light group) or 0 h of light and 24 h of darkness (0L:24D, dark group) for 18 D, with 2 replicates for each treatment. The results showed hatch time, time reaching 90% hatch, and average hatch time were significantly shorter among the 4 strains in the light group (P < 0.01). In addition, hatch window and peak hatching period were not extended by the green light stimulation (P > 0.05). There was no significant difference in hatchability of fertile eggs, chick weight/egg weight, or chick quality among the 4-strain eggs between the light group and dark group (P > 0.05). There was no difference (P > 0.05) in posthatch BW between different light treatments of the 3 strains (White Leghorn, Columbia Rock, and Barred Rock), whereas the BW of Rhode Island Red was higher in light group than that of the dark group at 8 to 12 wk of age (P < 0.05) and the difference disappeared from week 14. The results demonstrate that 12L:12D monochromatic green light stimulation during embryogenesis shortens the hatching time with no negative effects on hatching and posthatch performance. These effects were consistent among the 4 layer strains.

Tauroursodeoxycholic acid acts via TGR5 receptor to facilitate DNA damage repair and improve early porcine embryo development.

DNA damage associated with assisted reproductive technologies is an important factor affecting gamete fertility and embryo development. Activation of the TGR5 receptor by tauroursodeoxycholic acid (TUDCA) has been shown to reduce endoplasmic reticulum (ER) stress in embryos; however, its effect on genome damage responses (GDR) activation to facilitate DNA damage repair has not been examined. This study aimed to investigate the effect of TUDCA on DNA damage repair and embryo development. In a porcine model of ultraviolet light (UV)-induced nuclear stress, TUDCA reduced DNA damage and ER stress in developing embryos, as measured by γH2AX and glucose-regulated protein 78 immunofluorescence, respectively. TUDCA was equally able to rescue early embryo development. No difference in total cell number, DNA damage, or percentage of apoptotic cells, measured by cleaved caspase 3 immunofluorescence, was noted in embryos that reached the blastocyst stage. Interestingly, Dicer-substrate short interfering RNA-mediated disruption of TGR5 signaling abrogated the beneficial effects of TUDCA on UV-treated embryos. Quantitative PCR analysis revealed activation of the GDR, through increased messenger RNA abundance of DNAPK, 53BP1, and DNA ligase IV, as well as the ER stress response, through increased spliced XBP1 and X-linked inhibitor of apoptosis. Results from this study demonstrated that TUDCA activates TGR5-mediated signaling to reduce DNA damage and improve embryo development after UV exposure.

Strong static magnetic field delayed the early development of zebrafish.

One of the major topics in magnetobiology is the biological effects of strong static magnetic field (SMF) on living organisms. However, there has been a paucity of the comprehensive study of the long-term effects of strong SMF on an animal's development. Here, we explored this question with zebrafish, an excellent model organism for developmental study. In our research, zebrafish eggs, just after fertilization, were exposed to a 9.0 T SMF for 24 h, the critical period of post-fertilization development from cleavage to segmentation. The effects of strong SMF exposure on the following developmental progress of zebrafish were studied until 6 days post-fertilization (dpf). Results showed that 9.0 T SMF exposure did not influence the survival or the general developmental scenario of zebrafish embryos. However, it slowed down the developmental pace of the whole animal, and the late developers would catch up with their control peers after the SMF was removed. We proposed a mechanical model and deduced that the development delaying effect was caused by the interference of SMF in microtubule and spindle positioning during mitosis, especially in early cleavages. Our research data provide insights into how strong SMF influences the developing organisms through basic physical interactions with intracellular macromolecules.

Adaptive thermal plasticity enhances sperm and egg performance in a model insect.

Rising and more variable global temperatures pose a challenge for biodiversity, with reproduction and fertility being especially sensitive to heat. Here, we assessed the potential for thermal adaptation in sperm and egg function using Tribolium flour beetles, a warm-temperate-tropical insect model. Following temperature increases through adult development, we found opposing gamete responses, with males producing shorter sperm and females laying larger eggs. Importantly, this gamete phenotypic plasticity was adaptive: thermal translocation experiments showed that both sperm and eggs produced in warmer conditions had superior reproductive performance in warmer environments, and vice versa for cooler production conditions and reproductive environments. In warmer environments, gamete plasticity enabled males to double their reproductive success, and females could increase offspring production by one-third. Our results reveal exciting potential for sensitive but vital traits within reproduction to handle increasing and more variable thermal regimes in the natural environment.

Novel breeding approach for Japanese flounder using atmosphere and room temperature plasma mutagenesis tool.

BACKGROUND: Artificial induction of mutagenesis is effective for genetic resource innovation and breeding. However, the traditional mutation methods for fish breeding are not convenient or safe for daily use. Hence, development of a simple, safe and effective mutagenesis method with a high mutation rate and applicability to multiple fish species, is needed. RESULTS: We reported the first successful mutagenesis in a marine aquaculture fish species, Japanese flounder, Paralichthys olivaceus, using a novel atmosphere and room temperature plasma (ARTP) mutagenesis tool. ARTP treatment time was optimized for the fertilized eggs and sperm, respectively. Eggs fertilized for 60 min were treated by ARTP with a radio-frequency power input of 120 W, and the ARTP treatment time was 25 min. Under an ARTP radio-frequency power input of 200 W, the optimal treatment time for sperm diluted with Ringer's solution by 1:40 v/v was 10 min. The ARTP-treated group presented differences in morphological traits such as body height, total length among individuals at day 90 after hatching. Whole-genome sequencing was used to reveal the mutation features of ARTP-treated individuals collected at day 120 after hatching. In total, 69.25Gb clean data were obtained from three controls and eight randomly selected ARTP-treated individuals, revealing 240,722 to 322,978 SNPs and 82,149 to 86,798 InDels located in 17,394~18,457 and 12,907~13,333 genes, respectively. The average mutation rate reached 0.064% at the genome level. Gene ontology clustering indicated that genes associated with cell components, binding function, catalytic activity, cellular process, metabolic process and biological regulation processes had higher mutation rates. CONCLUSIONS: ARTP mutagenesis is a useful method for breeding of fish species to accelerate the selection of economically important traits that would benefit the aquaculture industry, given the variety of mutations detected.

Effect of Temperature on the Killing of Opisthorchis viverrini Eggs In Vitro.

Contaminated liver fluke egg in the environment has led to the high prevalence of human opisthorchiasis associated with cholangiocarcinoma in Southeast Asia. To find the effective lessening methods of Opisthorchis viverrini eggs in the contaminated environment, we investigated the temperature conditions for killing of these trematode eggs in vitro. Numerous O. viverrini eggs were obtained in the proximal part of uteri of adult worms from experimental hamsters. Mature eggs with miracidium were allocated by experimental groups (2 control: positive and negative and 4 treatment: 50, 60, 70, and 80°C) with 0.85% saline, and treated by the experimental plan. Eggs in each experimental groups were observed under the confocal microscope after stain with Propidium Iodide (PI) to evaluate the effect of temperatures. Eggs in 70 and 80°C groups were all killed after over 10 min heated. Majority of eggs in 60°C (10, 15, and 30 min heated), 70 and 80°C (5 min heated) groups were inactivated. However in 50°C group, below half of eggs were to be killed in all time lapse (10, 15 and 30 min). In order to prevent O. viverrini infection and cholangiocarcinoma, direct treatment of sewage by heating at 70 or 80°C at least 10 min is essential. Therefore, treatment of O. viverrini eggs at a high temperature is a potential method for controlling egg contamination in sewage.

The impact of Anopheles gambiae egg storage for mass rearing and production success.

BACKGROUND: Mass rearing requires a large colony from which male individuals can be harvested for sterilization and release. Attention is needed when monitoring life parameters of the reared population, knowing that any variations within the target population would lead to mismatching between two populations. The aim of this study was to assess the impact of Anopheles gambiae sensu stricto (s.s.) egg storage on hatchability and life history traits. For each parameter, comparison was made between freshly laid and stored eggs in three densities (40, 80, 120 eggs). METHODS: Anopheles gambiae s.s. freshly laid eggs were collected from the Tropical Pesticide Research Institute (TPRI) insectary. Eggs to be stored were kept at - 20 °C for 10 min and then transferred to refrigerators at 4 °C for intervals of 5, 10, 15, 20, and 25 days. After respective storage days, the eggs were transferred from refrigerators to ambient temperature of (25 ± 2) °C for 24 h and then placed in incubators for 24 h. Thereafter eggs were hatched. The egg hatchability, emerged larvae development, larvae survival and emerged adult sex ratios were monitored. RESULTS: This study found that hatching rates decreased with increase in storage time. The difference was significant in eggs stored for 10 and 15 days (P < 0.05). There were no significant differences in hatching rates between An. gambiae eggs stored for 5 days and freshly hatched eggs (P > 0.05). Anopheles larvae development (L1 to pupae) was not significantly affected by storage time across all hatching densities. The study also found that larvae survival decreased with increase in egg storage time. However, there was no significant difference between larvae from freshly hatched eggs and those from eggs at 5 and 10 storage days (P > 0.05) but not for eggs stored for 15 days. Furthermore, there was a decrease in emerged adult males and increase in females relative to increased time of egg storage. The difference was significant (P < 0.05) at 15 storage days but not for eggs stored for 5 and 10 days (in triplicate densities). CONCLUSION: From this study it was concluded that storing An. gambiae eggs at 4 °C and 48 ± 2% relative humidity (RH) for 5 days is the optimal condition and time that did not affect egg hatching rates, larval development and survivorship and emerged adult mosquito sex ratio.

Investigation of temperature conditions in Swiss urban and suburban microhabitats for the overwintering suitability of diapausing Aedes albopictus eggs.

BACKGROUND: In Switzerland, the invasive Asian tiger mosquito, Aedes albopictus, is firmly established in the Canton of Ticino, south of the Alps. According to a large-scale distribution model developed in 2013, suitable climatic conditions for the establishment of Ae. albopictus north of the Alps are found in Basel and Geneva while Zurich appears to be characterized by winters currently being too cold for survival of diapausing eggs. However, the spatial resolution of large-scale distribution models might not be sufficient to detect particular climatic conditions existing in urban settings, such as the presence of microclimatic temperatures, which may positively influence the probability of diapausing eggs to overwinter. In order to investigate this, microclimatic monitoring of potential diapausing sites (i.e. catch basins) and external controls was performed in January 2017 in Ticino and within the cities of Basel, Geneva and Zurich. RESULTS: Mean January temperatures in catch basins of Basel, Geneva and Zurich were always higher than the -1 °C temperature threshold previously set for survival probability of diapausing eggs, while mean January temperatures were below -1 °C in several catch basins south of the Alps, where Ae. albopictus eggs currently overwinter. The catch basin absolute January daily minimum temperatures both south and north of the Alps were in general higher than the external control temperatures. Absolute January daily minimum temperatures in catch basins in Basel, Geneva and Zurich were always above -10 °C, indicating that diapausing Ae. albopictus eggs could potentially survive winter nights in urban areas north of the Alps. CONCLUSIONS: The findings confirmed previous conclusions that urban catch basins can provide favourable conditions for overwintering of diapausing eggs compared to more cold-exposed sites. The results confirmed the presence of suitable winter conditions for the establishment of Ae. albopictus in the cities of Basel and Geneva. In addition, the microclimate-scale analysis added new information compared to the previous large-scale prevision model by showing that also the city of Zurich could provide winter conditions suitable for the establishment of Ae. albopictus. This illustrates the importance of the resolution of climate data in using models to predict Ae. albopictus distribution.

Non-invasive perturbations of intracellular flow reveal physical principles of cell organization.

Recent advances in cell biology enable precise molecular perturbations. The spatiotemporal organization of cells and organisms, however, also depends on physical processes such as diffusion or cytoplasmic flows, and strategies to perturb physical transport inside cells are not yet available. Here, we demonstrate focused-light-induced cytoplasmic streaming (FLUCS). FLUCS is local, directional, dynamic, probe-free, physiological, and is even applicable through rigid egg shells or cell walls. We explain FLUCS via time-dependent modelling of thermoviscous flows. Using FLUCS, we demonstrate that cytoplasmic flows drive partitioning-defective protein (PAR) polarization in Caenorhabditis elegans zygotes, and that cortical flows are sufficient to transport PAR domains and invert PAR polarity. In addition, we find that asymmetric cell division is a binary decision based on gradually varying PAR polarization states. Furthermore, the use of FLUCS for active microrheology revealed a metabolically induced fluid-to-solid transition of the yeast cytoplasm. Our findings establish how a wide range of transport-dependent models of cellular organization become testable by FLUCS.

Thermodynamics of egg production, development and hatching in trematodes.

Temperature is a key factor influencing the rate of biological processes of ectothermic animals and is intrinsically linked to climate change. Trematode parasites may be potentially susceptible to temperature changes and, in order to develop a predictive framework of their response to climate change, large-scale analyses are needed. In particular, the biology of the egg of all species is at some time influenced by environmental conditions. The present study uses Arrhenius activation energy (E*), a common measure of temperature-mediated reaction rates, to analyse experimental data from the scientific literature on the effects of temperature on the production, development and hatching of trematode eggs. Egg production declines at high temperatures, with habitat-specific climatic factors determining the optimal thermal range. Egg development, as is typical of invertebrates, shows a simple response to temperature, with minimal differences between mid- (35-60°) and low-latitude (<35°) species. Egg hatching demonstrates variable thermodynamics with high E* values at low temperature ranges and thermostability at mid-temperatures, before declining at high temperature ranges, with wide thermostable zones being a common feature. Comparisons between development and hatching indicate that these two parameters demonstrate different thermodynamical responses. The significance of these results in furthering our understanding of trematode egg biology under natural conditions is discussed.

Photoprotective effect of coumarin and 3-hydroxycoumarin in sea urchin gametes and embryonic cells.

Ultraviolet radiation B (UVB) represents 5% of all solar UV radiation and chronic exposure can induce harmful biological responses, including skin cancer. Prospection of new drugs with photoprotective properties and less toxic effects is constant and natural products have been the main options in this field. Coumarins are a group of natural phenolic compounds that shows several pharmacological activities. The aim of present work was to investigate the effect of coumarin and six derivatives in sea urchin gametes and zygotes exposed to UVB. Embryonic development assay was used to monitor UVB embryotoxicity. Firstly, we demonstrated that coumarin inhibited first embryonic cell division from 5 µM (EC50 = 52.9 µM) and its derivatives showed an embryotoxic effect ten times higher. Then, gametes or zygotes were treated with coumarin compounds before or after UVB exposure (UVB doses ranged from 0.056 to 0.9 kJm(-2)). Pretreatment of gametes or zygotes with coumarin or 3-hydroxycoumarin (1 µM, both) decreased UVB embryotoxic effect. Protective effect of the compounds was observed only when cells were treated previous to UVB exposure. Coumarin derivatives 4-hydroxycoumarin, 6-hydroxycoumarin, 7-hydroxycoumarin, 6,7-dihydroxycoumarin and 6-methoxy-7-hydroxycoumarin did not exhibit photoprotective activity. Our data provides evidences that coumarin and 3-hydroxycoumarin can be a promising class of photoprotective drugs.

Phototoxicity of narrowband ultraviolet (UV) B (311 nm) compared with broadband UVB in the photo hen's egg test.

BACKGROUND: Broadband ultraviolet B (BB-UVB) is a well-established treatment option in dermatology. However, during the last decade BB-UVB has increasingly been replaced by narrowband UVB 311 nm (NB-UVB), especially in the therapy of psoriasis, atopic eczema and vitiligo. Several studies have indicated a better therapeutic response for almost all indications compared with BB-UVB. OBJECTIVES: The aim of our study was to investigate the phototoxic effects of NB-UVB in comparison with BB-UVB in vivo. METHODS: Therefore, we employed the photo hen's egg test (PHET), an established phototoxic model, based on the yolk sac blood vessel system of incubated hen's eggs. NB-UVB and BB-UVB dosages increasing from 30 up to 1200 mJ cm(-2) were applied on 17 test groups (each n = 12 eggs) and two unirradiated test groups served as controls. Twenty-four hours after irradiation we observed the following test parameters: lethality, membrane discoloration and haemorrhages. RESULTS: Following our results, the lethal half dose (LD50) was 60 and 720 mJ cm(-2) for BB-UVB and NB-UVB, respectively. These LD50 dosages provoked severe membrane discoloration and haemorrhaging. Summarizing our results, the LD50 of NB-UVB was 12-fold higher than BB-UVB. CONCLUSIONS: Interestingly, these findings are in good accordance with the literature, where the minimal erythema dose (MED) of NB-UVB in human skin is up to 14 times higher than the MED of BB-UVB. These results show that the PHET is a valid test model to evaluate the phototoxic effects of various UVB wavelengths. Moreover, our results indicate that regarding the investigation of phototoxic effects the PHET might serve as a model representative for human skin, which might reduce the extent of photoprovocation in humans in the future.

Effect of red light on the development and quality of mammalian embryos.

PURPOSE: To assess irradiance and total energy dose from different microscopes during the in-vitro embryonic developmental cycle in mouse and pig and to evaluate its effect on embryonic development and quality in pig. METHOD: Spectral scalar irradiance (380-1050 nm) was measured by a fiber-optic microsensor in the focal plane of a dissection microscope, an inverted microscope and a time-lapse incubation system. Furthermore, the effect of three different red light levels was tested in the time-lapse system on mouse zygotes for 5 days, and on porcine zona-intact and zona-free parthenogenetically activated (PA) embryos for 6 days. RESULTS: The time-lapse system used red light centered at 625 nm and with a lower irradiance level as compared to the white light irradiance levels on the dissection and inverted microscopes, which included more energetic radiation <550 nm. Even after 1000 times higher total energy dose of red light exposure in the time-lapse system, no significant difference was found neither in blastocyst development of mouse zygotes nor in blastocyst rates and total cell number of blastocysts of porcine PA embryos. CONCLUSIONS: Our results indicate that red light (625 nm, 0.34 W/m(2)) used in the time-lapse incubation system does not decrease the development and quality of blastocysts in both mouse zygotes and porcine PA embryos (both zona-intact and zona-free).

Peering beneath the surface: novel imaging techniques to noninvasively select gametes and embryos for ART.

Embryo imaging has long been a critical tool for in vitro fertilization laboratories, aiding in morphological assessment of embryos, which remains the primary tool for embryo selection. With the recent emergence of clinically applicable real-time imaging systems to assess embryo morphokinetics, a renewed interest has emerged regarding noninvasive methods to assess gamete and embryo development as a means of inferring quality. Several studies exist that utilize novel imaging techniques to visualize or quantify intracellular components of gametes and embryos with the intent of correlating localization of organelles or molecular constitution with quality or outcome. However, the safety of these approaches varies due to the potential detrimental impact of light exposure or other variables. Along with complexity of equipment and cost, these drawbacks currently limit clinical application of these novel microscopes and imaging techniques. However, as evidenced by clinical incorporation of some real-time imaging devices as well as use of polarized microscopy, some of these imaging approaches may prove to be useful. This review summarizes the existing literature on novel imaging approaches utilized to examine gametes and embryos. Refinement of some of these imaging systems may permit clinical application and serve as a means to offer new, noninvasive selection tools to improve outcomes for various assisted reproductive technology procedures.

[Embryonic development of whitefishes (Coregonidae) as representatives of the "pagophilous" ecological group].

Studies of reproduction and embryonic development in six species of coregonid fishes have revealed the possibility of their fertilized eggs to develop normally while being embedded in the ice of a spawning water body (optionally). Such ability is facilitated by extremely low respiratory activity of embryos at early stages of embryogenesis (from the stage of fission to the stage of organogenesis). Low level of oxygen consumption and carbon dioxide emission is an adaptation to low diffusion gas permeability of the ice. The main factor controlling the rate of coregonids embryonic development is not temperature, but intensity and periodicity of insolation. Without the sunlight--an obligatory external factor--normal development is just not possible. Under experimental conditions, when developing in the water at near zero temperature or in the ice, normal morphogenesis of Arctic cisco and Sevan whitefish embryos was observed at the illumination of 50-300 lux. Hemoproteid cytochrome beta560, the pigment that has been discovered in water-soluble part of coregonids oocyte yolk and is treated as a biochemical marker for eggs of the family Coregonidae, in all likelihood performs protective (antioxidant) functions preventing spontaneous oxidation of embryo's fatty inclusions. Under the oxygen shortage inside the ice envelope, cytochrome beta560 probably sets conditions for oxidation processes of embryo's tissue respiration. Spherome, being kept till the time of hatching, acts as a temporary hydrostatic organ and ensures larvae buoyancy at the stage of postembryonic metamorphosis. It also serves as an energy store after downstream migration of larvae from the spawning areas till their shift to exogenous feeding on zooplankton. Conforming to ecological traits of reproduction and development, and also to revealed morphogenetic, physiological, and biochemical features, it is proposed to ascribe all of the currently known 26 species of whitefishes to "pagophilous" ecological group.

Production of androgenetic diploid loach by cold-shock of eggs fertilized with diploid sperm.

Diploid androgenotes were produced without egg irradiation in the loach, Misgurnus anguillicaudatus. Eggs of wild-type diploid females were fertilized with diploid sperm of a neo-tetraploid male and then cold-shock treated at 3 °C (range, ±0.5 °C) for 30 minutes just after fertilization to eliminate the female nucleus. After hatching, ploidy status of the hatched larvae was analyzed by flow cytometry, which revealed putative diploid androgenotes as well as larvae possessing other ploidies. Five independent microsatellite DNA markers were genotyped to confirm all-male inheritance of the resultant diploid larvae. The mean ± SD yield rate of diploid androgenetic larvae to total eggs used was 12.29 ± 3.25% in the cold-shock group and 22.23 ± 13.42% in the UV-irradiated group (P > 0.05). No diploid androgenetic larvae were detected in the intact control group. To our knowledge, this is the first report demonstrating successful induction of diploid androgenotes without egg irradiation in fish.

Stable and fluctuating temperature effects on the development rate and survival of two malaria vectors, Anopheles arabiensis and Anopheles funestus.

BACKGROUND: Understanding the biology of malaria vector mosquitoes is crucial to understanding many aspects of the disease, including control and future outcomes. The development rates and survival of two Afrotropical malaria vectors, Anopheles arabiensis and Anopheles funestus, are investigated here under conditions of constant and fluctuating temperatures. These data can provide a good starting point for modelling population level consequences of temperature change associated with climate change. For comparative purposes, these data were considered explicitly in the context of those available for the third African malaria vector, Anopheles gambiae. METHODS: Twenty five replicates of 20-30 eggs were placed at nine constant and two fluctuating temperatures for development rate experiments and survival estimates. Various developmental parameters were estimated from the data, using standard approaches. RESULTS: Lower development threshold (LDT) for both species was estimated at 13-14°C. Anopheles arabiensis developed consistently faster than An. funestus. Optimum temperature (Topt) and development rate at this temperature (µmax) differed significantly between species for overall development and larval development. However, Topt and µmax for pupal development did not differ significantly between species. Development rate and survival of An. funestus was negatively influenced by fluctuating temperatures. By contrast, development rate of An. arabiensis at fluctuating temperatures either did not differ from constant temperatures or was significantly faster. Survival of this species declined by c. 10% at the 15°C to 35°C fluctuating temperature regime, but was not significantly different between the constant 25°C and the fluctuating 20°C to 30°C treatment. By comparison, previous data for An. gambiae indicated fastest development at a constant temperature of 28°C and highest survival at 24°C. CONCLUSIONS: The three most important African malaria vectors all differ significantly in development rates and survival under different temperature treatments, in keeping with known distribution data, though differences among M and S molecular forms of An. gambiae likely complicate the picture. Increasing temperatures associated with climate change favour all three species, but fluctuations in temperatures are detrimental to An. funestus and may also be for An. gambiae. This may have significant implications for disease burden in areas where each species is the main malaria vector.