Despite recent reports regarding the biology of cytosine methylation in Schistosoma mansoni, the impact of the regulatory machinery remains unclear in diverse platyhelminthes. This ambiguity is reinforced by discoveries of DNA methyltransferase 2 (DNMT2)-only organisms and the substrate specificity of DNMT2 preferential to RNA molecules. Here, we characterized a novel DNA methyltransferase, named CsDNMT2, in a liver fluke Clonorchis sinensis. The protein exhibited structural properties conserved in other members of the DNMT2 family. The native and recombinant CsDNMT2 exhibited considerable enzymatic activity on DNA. The spatiotemporal expression of CsDNMT2 mirrored that of 5-methylcytosine (5 mC), both of which were elevated in the C. sinensis eggs. However, CsDNMT2 and 5 mC were marginally detected in other histological regions of C. sinensis adults including ovaries and seminal receptacle. The methylation site seemed not related to genomic loci occupied by progenies of an active long-terminal-repeat retrotransposon. Taken together, our data strongly suggest that C. sinensis has preserved the functional DNA methylation machinery and that DNMT2 acts as a genuine alternative to DNMT1/DNMT3 to methylate DNA in the DNMT2-only organism. The epigenetic regulation would target functional genes primarily involved in the formation and/or maturation of eggs, rather than retrotransposons.

The duration of brood development in the introduced crab, Hemigrapsus takanoi in the Oosterschelde, The Netherlands, was compared at three different water temperatures. At 12, 18 and 24°C the females took an average of 32, 11 and 8 days respectively to lay eggs, which took 86, 28 and 18 days respectively to complete development. Five stages of development were identified, with each brood stage comprising a similar proportion of the duration time at different temperatures. The duration of each brood stage was also somewhat proportional to the number of females found carrying each brood stage in the field at the beginning of the breeding season. There appears to be a trigger for the breeding season in H. takanoi in the field at around 15°C above which ovary development begins. The results suggest that an increase in water temperature as a result of climate change may result in an increased net reproductive rate in H. takanoi due to earlier onset of the breeding season and increased number of broods per inter-moult period resulting in population growth. Increased temperatures may therefore lead to increased invasiveness of H. takanoi where it is already present, and range extension into locations where its establishment is currently excluded by unsuitable temperature.

The effect of temperature on brood development was investigated for three intertidal hymenosomatid crabs: Halicarcinus cookii, H. varius and H. innominatus in Kaikoura, New Zealand. The duration of brood incubation decreased as temperature increased, as did the interbrood period. The duration of each stage of brood development also decreased with increased temperature, but the proportion of total incubation time for each stage remained relatively similar at different temperatures. Hymenosomatid crabs have determinate growth, but moult to maturity at different sizes, thereafter devoting most of their energy to reproduction. The number of broods a female could carry in her lifetime was estimated for each species. Halicarcinus cookii was estimated to be able to produce eight complete broods of 1146 eggs per lifetime, H. varius was estimated to be able to produce seven complete broods of 1051 eggs per lifetime and H. innominatus was estimated to be able to produce six complete broods of 1081 eggs per life time. With the predicted global temperature rise of 2°C in the next 50 years, the authors estimate that, for all three species, a female could produce one extra brood per lifetime (a 10–15% increase in fecundity depending on species), even more if crabs reach maturity faster, potentially leading to a significant population increase.

The development and mortality of the eggs of Ancylostoma duodenale and Necator americanus in distilled water were monitored over a range of temperatures between 15 and 35°C. Egg demography was examined within the context of a fourparameter mathematical model of development and mortality. Over the range of temperatures studied, egg mortality (μ) was an increasing exponential function of temperature (T) measured in degrees Celsius. A single model adequately described the mortality of both species (ln [μ] = 0·041*T − 6·87). The minimum time (τ) to hatching was consistently less for A. duodenale (ln [1/T] = −(0·79+53·05*[l/T]) than N. americanus (ln [1/T] = −(0·99 + 53·05*[l/T])). The hatching rate [σ when t > τ) was an increasing function of time and temperature in both cases, but the precise functional relationship was species specific.

The root-feeding flea beetle Longitarsus bethae Savini & Escalona, was introduced into South Africa as a candidate biological control agent for the noxious and invasive weed, Lantana camara L. As part of the study to predict the beetles' survival in its new range, the influence of climatic conditions on its egg development and reproductive performance were investigated in the laboratory. The threshold temperature (T°) and degree-days (DD) required for egg hatch were determined after exposing the eggs to various constant temperatures (12, 17, 22, 27 and 32°C) in separate growth chambers. The DD required for egg hatch was 178.6, and the temperature threshold required for egg hatch was 11.3°C. Survival of eggs varied from 27 to 56% at 32 and 17°C, respectively, and was optimum between 17 and 25°C. Oviposition was examined under high and low relative humidity (RH) regimes while egg hatch was determined at six RH levels, each maintained in a separate controlled growth chamber set at a constant temperature (25°C). Whilst RH had no influence on oviposition, eggs were highly susceptible to aridity, and continuous exposure to relative humidity below 63% for more than three days was wholly lethal at 25°C. Optimum egg hatch occurred at RH between 85 and 95% for up to 12 days. The effect of day length on oviposition and subsequent egg hatch was investigated under two photoperiod regimes. Neither oviposition nor subsequent egg hatch was influenced by photoperiod. The knowledge obtained will be useful for mass rearing as well as field release programmes for L. bethae.

The influence of drought on egg development, survival and hatchling weight was studied in the solitary phase of Locusta migratoria migratorioides (Reiche and Fairmaire). There was retarding effect of drought on the rate of egg development. The level of egg survival was lowered during prolonged drought, which also led to a decline in hatchling weight. Also the rate of egg survival was lower in eggpods experiencing longer incubation period.

Laboratory studies were conducted on the role of flood on egg development, survival and hatchling weight in the solitary phase of Locusta migratoria migratorioides (Reiche and Fairmaire). Egg development is retarded by flood, particularly if the eggs develop for a long period in ordinary moist soil prior to flooding. Floods are also detrimental to egg survival; eggs that are flooded immediately after oviposition and those flooded when they are about to hatch are particularly susceptible. Egg age at flooding is a major determining factor of hatchling weight; heavier hatchlings are produced if eggs develop in ordinary moist soil for longer, rather than shorter, period before being flooded. Length of flooding period has no apparent effect on hatchling weight.