African Annual Fish Research Articles

Embryo ecology: Developmental synchrony and asynchrony in the embryonic development of wild annual fish populations.

Embryo–environment interactions are of paramount importance during the development of all organisms, and impacts during this period can echo far into later stages of ontogeny. African annual fish of the genus Nothobranchius live in temporary pools and their eggs survive the dry season in the dry bottom substrate of the pools by entering a facultative developmental arrest termed diapause. Uniquely among animals, the embryos (encased in eggs) may enter diapause at three different developmental stages. Such a system allows for the potential to employ different regulation mechanisms for each diapause. We sampled multiple Nothobranchius embryo banks across the progressing season, species, and populations. We present important baseline field data and examine the role of environmental regulation in the embryonic development of this unique system. We describe the course of embryo development in the wild and find it to be very different from the typical development under laboratory conditions. Development across the embryo banks was synchronized within and across the sampled populations with all embryos entering diapause I during the rainy season and diapause II during the dry season. Asynchrony occurred at transient phases of the habitat, during the process of habitat desiccation, and at the end of the dry season. Our findings reveal the significance of environmental conditions in the serial character of the annual fish diapauses.

Individual-level pace-of-life syndromes in annual killifish are mediated by intersexual and interspecific differences

Pace-of-life syndromes (POLS) describe covariations between life history (such as growth rate and age at maturity), behaviour (e.g. activity or boldness) and physiology (e.g. metabolic rate) along an axis from fast to slow lifestyles. This powerful concept can be applicable at a range of scales, from broad interspecific contrasts to the individual intra-population level, though its generality has recently been questioned. Using two species of African annual fishes with fast lifestyles, we tested how individual-level POLS covary between the sexes and contrasting social environments. Measuring three key metabolic parameters (standard metabolic rate, SMR; maximum metabolic rate, MMR; and aerobic scope, AS), we found extensive variation between species and sexes in the expression of POLS. Social environment affected individual metabolic traits, but not their covariation with behaviour and life history traits. In accordance with the POLS prediction, we observed a positive association between MMR, AS and boldness, and a negative association between MMR, AS and lifespan in Nothobranchius orthonotus males, although trait covariations were opposite in N. orthonotus females. In Nothobranchius pienaari, we confirmed the predicted negative correlation between SMR and lifespan which was not sex-specific. Contrary to POLS predictions, we observed a negative correlation between SMR and boldness in N. pienaari. Finally, there was no link between activity levels or size at maturity and metabolic traits in either species. Overall, we demonstrated limited support for POLS predictions, but found that specific pace-of-life trait associations were mediated by both interspecific and intersexual differences.

Genetic diversity of a widespread annual killifish from coastal Tanzania

BackgroundAfrican annual killifishes (Nothobranchius spp.) are adapted to seasonally desiccating habitats (ephemeral pools), surviving dry periods as dormant eggs. Given their peculiar life history, geographic aspects of their diversity uniquely combine patterns typical for freshwater taxa (river basin structure and elevation gradient) and terrestrial animals (rivers acting as major dispersal barriers). However, our current knowledge on fine-scale inter-specific and intra-specific genetic diversity of African annual fish is limited to a single, particularly dry region of their distribution (subtropical Mozambique). Using a widespread annual killifish from coastal Tanzania and Kenya, we tested whether the same pattern of genetic divergence pertains to a wet equatorial region in the centre of Nothobranchius distribution.ResultsIn populations of Nothobranchius melanospilus species group across its range, we genotyped a part of mitochondrial cytochrome oxidase subunit 1 (COI) gene (83 individuals from 22 populations) and 10 nuclear microsatellite markers (251 individuals from 16 populations). We found five lineages with a clear phylogeographic structure but frequent secondary contact. Mitochondrial lineages were largely congruent with main population genetic clusters identified on microsatellite markers. In the upper Wami basin, populations are isolated as a putative Nothobranchius prognathus, but include also a population from a periphery of the middle Ruvu basin. Other four lineages (including putative Nothobranchius kwalensis) coexisted in secondary contact zones, but possessed clear spatial pattern. Main river channels did not form apparent barriers to dispersal. The most widespread lineage had strong signal of recent population expansion.ConclusionsWe conclude that dispersal of a Nothobranchius species from a wet part of the genus distribution (tropical lowland) is not constrained by main river channels and closely related lineages frequently coexist in secondary contact zones. We also demonstrate contemporary connection between the Ruvu and Rufiji river basins. Our data do not provide genetic support for existence of recently described cryptic species from N. melanospilus complex, but cannot resolve this issue.

Hatching date variability in wild populations of four coexisting species of African annual fishes.

Hatching is modulated by a combination of intrinsic and extrinsic factors. Annual killifish are adapted to complete their entire life cycle in annually desiccating habitats. Spending most of their life in the embryonic stage, they have evolved adaptations to survive desiccated conditions and match their hatching with the unpredictable onset of the aquatic phase of the pool. We examined spatial and temporal synchrony of hatching in natural populations of four species of African annual killifish (genus Nothobranchius). We compared differences and variability in hatching dates among years, regions, pools, and species and matched them with data on inundations of individual pools. Inundations typically coincided with peak rainfall in early January. We found considerable spatial and temporal synchrony in 1 year, but less synchrony in the other 2 years. Hatching generally occurred 0-20 days after inundation; fish at most sites hatched synchronously (<1 week) but some sites showed protracted hatching or two age cohorts. One species tended to hatch earlier than the other three. We suggest that hatching of annual killifish in the wild is a result of the interplay between environmental conditions and individual predisposition to respond to threshold environmental cues, ensuring effective bet-hedging against unpredictable inundation. Developmental Dynamics 246:827-837, 2017. © 2017 Wiley Periodicals, Inc.

Repeated intraspecific divergence in life span and aging of African annual fishes along an aridity gradient.

Life span and aging are substantially modified by natural selection. Across species, higher extrinsic (environmentally related) mortality (and hence shorter life expectancy) selects for the evolution of more rapid aging. However, among populations within species, high extrinsic mortality can lead to extended life span and slower aging as a consequence of condition-dependent survival. Using within-species contrasts of eight natural populations of Nothobranchius fishes in common garden experiments, we demonstrate that populations originating from dry regions (with short life expectancy) had shorter intrinsic life spans and a greater increase in mortality with age, more pronounced cellular and physiological deterioration (oxidative damage, tumor load), and a faster decline in fertility than populations from wetter regions. This parallel intraspecific divergence in life span and aging was not associated with divergence in early life history (rapid growth, maturation) or pace-of-life syndrome (high metabolic rates, active behavior). Variability across four study species suggests that a combination of different aging and life-history traits conformed with or contradicted the predictions for each species. These findings demonstrate that variation in life span and functional decline among natural populations are linked, genetically underpinned, and can evolve relatively rapidly.

Female fecundity traits in wild populations of African annual fish: the role of the aridity gradient.

The evolution of life history is shaped by life expectancy. Life‐history traits coevolve, and optimal states for particular traits are constrained by trade‐offs with other life‐history traits. Life histories contrast among species, but may also diverge intraspecifically, at the level of populations. We studied the evolution of female reproductive allocation strategy, using natural populations of two sympatric species of African annual fishes, Nothobranchius furzeri and Nothobranchius orthonotus. These species inhabit pools in the Mozambican savanna that are formed in the rainy season and persist for only 2–10 months. Using 207 female N. furzeri from 11 populations and 243 female N. orthonotus from 14 populations, we tested the effects of genetic background (intraspecific lineage) and life expectancy (position on the aridity gradient determining maximum duration of their temporary habitat) on female fecundity traits. First, we found that variation in female body mass was small within populations, but varied considerably among populations. Second, we found that fecundity was largely defined by female body mass and that females spawned most of their eggs in the morning. Third, we found that the trade‐off between egg size and egg number varied among lineages of N. furzeri and this outcome has been confirmed by data from two separate years. Overall, we demonstrate that local conditions were important determinants for Nothobranchius growth and fecundity and that eggs size in arid region was less limited by female fecundity than in humid region.

From the bush to the bench: the annual Nothobranchius fishes as a new model system in biology.

African annual fishes from the genus Nothobranchius are small teleosts that inhabit temporary water bodies subject to annual desiccation due to the alternation of the monsoon seasons. Given their unique biology, these fish have emerged as a model taxon in several biological disciplines. Their increasing popularity stems from the extremely short lifespan that is the result of their specific life-history adaptations and is retained under laboratory conditions. Nothobranchius furzeri, the most popular laboratory species, is the vertebrate species with the shortest lifespan recorded in captivity. In the laboratory, adults of different Nothobranchius species and populations live between 3 and 18 months and, notably, there is a negative correlation between the captive lifespan of a species and the aridity of their habitat. Their short lifespan is coupled to rapid age-dependent functional decline and expression of cellular and molecular changes comparable to those observed in other vertebrates, including humans. The recent development of transgenesis in this species makes it possible to insert specific constructs into their genome, and the establishment of transgenic lines is facilitated by their very rapid generation time, which can be as short as 1 month. This makes Nothobranchius species particularly suited for investigating biological and molecular aspects of ageing and ageing-associated dysfunctions. At the same time, they also represent a unique model taxon to investigate the evolution of life-history adaptations and their genetic architecture. We review their natural history, including phylogenetic relationships, distribution in relation to habitat conditions and natural selection for differential longevity, population structure and demography, and life cycle with emphasis on diapause that may occur at three stages during embryonic development. We further critically evaluate their use as a laboratory model for understanding the evolution of a rapid ageing rate and its consequences for other life-history traits, for cellular, molecular and integrative traits associated with the ageing process, high incidence of neoplasias, their utility for genome-wide gene-expression studies, and as a model for quantitative genetics. We summarize recent achievements in fostering Nothobranchius species as a widely applicable model system, including an annotated transcriptome, successful transgenesis, and existence of viable inbred lines. We compare the conditions they experience in the wild and in captivity and suggest that they are an ideal taxon to investigate natural genetic variation in a laboratory setting. We conclude that Nothobranchius species - and N. furzeri in particular - could become a unique model taxon that bridges interests in ecological and biomedical research. We hope that a conceptual and methodological integration of these two branches of biology will provide important new insights.

The strange case of East African annual fishes: aridification correlates with diversification for a savannah aquatic group?

BackgroundAnnual Nothobranchius fishes are distributed in East and Southern Africa and inhabit ephemeral pools filled during the monsoon season. Nothobranchius show extreme life-history adaptations: embryos survive by entering diapause and they are the vertebrates with the fastest maturation and the shortest lifespan. The distribution of Nothobranchius overlaps with the East Africa Rift System. The geological and paleoclimatic history of this region is known in detail: in particular, aridification of East Africa and expansion of grassland habitats started 8 Mya and three humid periods between 3 and 1 Mya are superimposed on the longer-term aridification. These climatic oscillations are thought to have shaped evolution of savannah African mammals. We reconstructed the phylogeny of Nothobranchius and dated the different stages of diversification in relation to these paleoclimatic events.ResultsWe sequenced one mitochondrial locus and five nuclear loci in 63 specimens and obtained a robust phylogeny. Nothobranchius can be divided in four geographically separated clades whose boundaries largely correspond to the East Africa Rift system. Statistical analysis of dispersal and vicariance identifies a Nilo-Sudan origin with southwards dispersion and confirmed that these four clades are the result of vicariance events In the absence of fossil Nothobranchius, molecular clock was calibrated using more distant outgroups (secondary calibration). This method estimates the age of the Nothobranchius genus to be 8.3 (6.0 – 10.7) My and the separation of the four clades 4.8 (2.7-7.0) Mya. Diversification within the clades was estimated to have started ~3 Mya and most species pairs were estimated to have an age of 0.5-1 My.ConclusionsThe mechanism of Nothobranchius diversification was allopatric and driven by geographic isolation. We propose a scenario where diversification of Nothobranchius started in rough coincidence with aridification of East Africa, establishment of grassland habitats and the appearance of the typical African bovid fauna of the savannah. Although confidence intervals for the estimated ages of the four Nothobranchius clades are quite large, this scenario is compatible with the biology of extant Nothobranchius that are critically dependent on savannah habitats. Therefore, Nothobranchius diversification might have been shaped by the same paleoclimatic events that shaped African ungulate evolution.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-014-0210-3) contains supplementary material, which is available to authorized users.

Highly plastic resource allocation to growth and reproduction in females of an African annual fish

AbstractPhenotypic plasticity is an important mechanism to maximise fitness in unpredictable environments by fine‐tuning phenotypes to a specific environmental setting. We used Nothobranchius furzeri, an African annual fish from temporary pools with erratic changes in habitat condition, to study changes in the allocation to growth and reproduction and to test the key trade‐off between egg size and number. In an experimental setting, we quantitatively varied ration at two levels and over two time periods, including temporal switches in ration level. As predicted, female N. furzeri possessed the capacity for compensatory growth, which surprisingly came with no longer‐term cost to fecundity. Females responded strongly to ration manipulation, with a pronounced decrease in fecundity associated with a low ration, even after accounting for body mass. Due to the unpredictability of offspring environment, we expected no adaptive change in oocyte size. However, females responded to the quality of their environment in accordance with an adaptive maternal effect, with females receiving a low ration producing larger eggs. Further, a switch in ration size in either direction was associated with a decrease in egg size. There was a trade‐off between egg size and number in half of the treatments, but high variability in egg size among females made the relationship complex. Overall, N. furzeri females demonstrated high plasticity in both growth rate and fecundity parameters. Females appear able to track and respond adaptively to unpredictable changes in food availability in their environment.

Female bias in the adult sex ratio of African annual fishes: interspecific differences, seasonal trends and environmental predictors

Many populations have consistently biased adult sex ratios with important demographic and evolutionary consequences. However, geographical variation, the mechanisms, temporal dynamics and predictors of biased sex ratios are notoriously difficult to explain. We studied 334 wild populations of four species of African annual fish (Nothobranchius furzeri, N. kadleci, N. orthonotus, N. rachovii) across their ranges to compare their adult sex ratio, its seasonal dynamics, interpopulation variation and environmental predictors. Nothobranchius populations comprise a single age cohort and inhabit discrete isolated pools, with wide-ranging environmental conditions (habitat size, water turbidity, structural complexity, predators), making them ideal to study adult sex ratio variation. In captivity adult sex ratios were equal. In natural populations, adult sex ratios were biased 1:2 toward females in three study species while N. kadleci had sex ratios at unity. There was a decline in the proportion of males with age in one species, but not in the other species, implying most severe male mortality early after maturation, declining later perhaps with a decrease in male abundance. In general, the populations at vegetated sites had relatively more males than populations at sites with turbid water and little vegetation. Selective avian predation on brightly coloured male fish likely contributed to female dominance and vegetation cover may have protected males from birds. In addition, an aquatic predator, a large belastomid hemipteran, decreased the proportion of males in populations, possibly due to greater male activity rather than conspicuous colouration. Alternative explanations for a sex ratio bias, stemming from male–male contests for matings, are discussed. We conclude that the effect of environmental conditions on adult sex ratio varies dramatically even in closely related and ecologically similar sympatric species. Therefore, difficulties in explaining the ecological predictors of sex ratio biases are likely due to high stochasticity rather than limited sample size.

Rapid growth, early maturation and short generation time in African annual fishes

BackgroundExtreme environmental conditions can give rise to extreme adaptations. We document growth, sexual maturation and fecundity in two species of African annual fish inhabiting temporary savanna pools.ResultsNothobranchius kadleci started to reproduce at the age of 17 days and size of 31 mm and Nothobranchius furzeri at 18 days and 32 mm. All four study populations demonstrated rapid growth rates of up to 2.72 mm/day (23.4% of their total length). Both species may produce diapausing embryos or embryos that are able to hatch in as few as 15 days, resulting in a minimum generation time as short as only one month. Incubation on the surface of damp peat moss results in high embryo survival (73%) and a high proportion of rapidly developing embryos (58%) that skip diapauses and hatch in less than 30 days. We further demonstrated that rapid growth and maturation do not compromise subsequent fecundity.ConclusionsOur data suggest that both species have the most rapid sexual maturation and minimum generation time of any vertebrate species, and that rapid maturity does not involve paedogenesis.

Trophic niche partitioning in communities of African annual fish: evidence from stable isotopes

Annual killifish of the genus Nothobranchius often co-occur in temporary savannah pools. Their space- and time-limited environment does not allow for any substantial habitat or temporal segregation. Coexisting species are therefore predicted to have well separated trophic niches to avoid intense food competition. Although in a previous “snapshot” study using stomach content analysis (SCA), the trophic niches of three sympatric species (N. furzeri, N. orthonotus, and N. pienaari) were found to vary among species, the difference was relatively weak and inconsistent across different sites. Here, we used the time-integrative capacity of stable isotope analysis to test whether the trophic niches of sympatric Mozambican Nothobranchius are more distinct over a long-term period. Analysis of carbon and nitrogen stable isotopes separated the trophic niche and trophic position of N. pienaari but failed to find any difference between N. furzeri/N. kadleci and N. orthonotus. No segregation was found at the sites with low prey diversity. In contrast, SCA identified N. orthonotus as the species with the most distinct trophic niche. We discuss the effect of prey diversity and different sensitivities of stomach content and stable isotope analysis in general and conclude that the trophic niches of the three sympatric Nothobranchius species are well separated.

The transcript catalogue of the short-lived fish Nothobranchius furzeri provides insights into age-dependent changes of mRNA levels

BackgroundThe African annual fish Nothobranchius furzeri has over recent years been established as a model species for ageing-related studies. This is mainly based on its exceptionally short lifespan and the presence of typical characteristics of vertebrate ageing. To substantiate its role as an alternative vertebrate ageing model, a transcript catalogue is needed, which can serve e.g. as basis for identifying ageing-related genes.ResultsTo build the N. furzeri transcript catalogue, thirteen cDNA libraries were sequenced using Sanger, 454/Roche and Solexa/Illumina technologies yielding about 39 Gb. In total, 19,875 protein-coding genes were identified and annotated. Of these, 71% are represented by at least one transcript contig with a complete coding sequence. Further, transcript levels of young and old fish of the strains GRZ and MZM-0403, which differ in lifespan by twofold, were studied by RNA-seq. In skin and brain, 85 differentially expressed genes were detected; these have a role in cell cycle control and proliferation, inflammation and tissue maintenance. An RNA-seq experiment for zebrafish skin confirmed the ageing-related relevance of the findings in N. furzeri. Notably, analyses of transcript levels between zebrafish and N. furzeri but also between N. furzeri strains differed largely, suggesting that ageing is accelerated in the short-lived N. furzeri strain GRZ compared to the longer-lived strain MZM-0403.ConclusionsWe provide a comprehensive, annotated N. furzeri transcript catalogue and a first transcriptome-wide insight into N. furzeri ageing. This data will serve as a basis for future functional studies of ageing-related genes.

Mapping of quantitative trait loci controlling lifespan in the short‐lived fish Nothobranchius furzeri– a new vertebrate model for age research

The African annual fish Nothobranchius furzeri emerged as a new model for age research over recent years. Nothobranchius furzeri show an exceptionally short lifespan, age-dependent cognitive/behavioral decline, expression of age-related biomarkers, and susceptibility to lifespan manipulation. In addition, laboratory strains differ largely in lifespan. Here, we set out to study the genetics of lifespan determination. We crossed a short- to a long-lived strain, recorded lifespan, and established polymorphic markers. On the basis of genotypes of 411 marker loci in 404 F2 progeny, we built a genetic map comprising 355 markers at an average spacing of 5.5 cM, 22 linkage groups (LGs) and 1965 cM. By combining marker data with lifespan values, we identified one genome-wide highly significant quantitative trait locus (QTL) on LG 9 (P < 0.01), which explained 11.3% of the F2 lifespan variance, and three suggestive QTLs on LG 11, 14, and 17. We characterized the highly significant QTL by synteny analysis, because a genome sequence of N. furzeri was not available. We located the syntenic region on medaka chromosome 5, identified candidate genes, and performed fine mapping, resulting in a c. 40% reduction of the initial 95% confidence interval. We show both that lifespan determination in N. furzeri is polygenic, and that candidate gene detection is easily feasible by cross-species analysis. Our work provides first results on the way to identify loci controlling lifespan in N. furzeri and illustrates the potential of this vertebrate species as a genetic model for age research.

Reproductive isolating barriers between colour-differentiated populations of an African annual killifish, Nothobranchius korthausae (Cyprinodontiformes)

Allopatric populations separated by vicariance events are expected to evolve reproductive isolating mechanisms as a result of disparate selection pressures and genetic drift. The appearance of reproductive isolating mechanisms may vary across taxa with differences in the opportunity for mate choice, and may be asymmetrical. In addition, premating barriers may be affected by individual mating experience. We used choice and no-choice experiments to investigate reproductive isolation between two allopatric (island and mainland) and colour-differentiated populations of an African annual fish, Nothobranchius korthausae. Assortative mating under experimental conditions was limited and asymmetrical. Preference for sympatric males was only expressed in nonvirgin females from one population. Virgin fish from both populations mated indiscriminately. No difference in the number of eggs laid, fertilization rate and hatching success was detected in no-choice experiments. All mating combinations produced viable offspring and no postmating barriers were detected in terms of the performance and fertility of F1 hybrids. Overall, we found little evidence for significant reproductive isolation, which is in contrast with the related killifish taxa in which assortative mating can be strong, even among allopatric populations with no colour differentiation. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100, 62–72.

Potential negative impacts and low effectiveness in the use of African annual killifish in the biocontrol of aquatic mosquito larvae in temporary water bodies

Commentary and discussion on a recent paper promoting the use of Nothobranchius guentheri, a small African annual fish from the Island of Zanzibar as a tool to control mosquito larvae in temporary bodies of freshwater throughout Africa is presented.Arguments on major points; (1) expected low success of annual fish introductions, (2) low success of mosquito control in the field, (3) ecological threats, and (4) ethical issues are detailed.Despite serious problems with mosquito-borne diseases in tropical Africa and elsewhere, we encourage responsible means of biological control of parasite vectors. We show that effectiveness of Nothobranchius translocations is low (the previous attempts failed), likelihood of effective mosquito larvae control under field condition is negligible and ecological threats from Nothobranchius translocations from within and outside the naturally occurring range are serious. We advocate against the proposed next step of the project, i.e. field trials in Tanzania.

Cell behavior during early development in the South American annual fishes of the genus Cynolebias

Living embryos of three species of South American annual fishes, Cynolebias constanciae, C. nigripinnis, and C. whitei, were observed from fertilization through the 10-somite stage. A description of normal stages of development applicable to all three species of Cynolebias is presented. Cleavage (stages 1-10) is meroblastic and produces a typical teleost blastoderm. Following cleavage (stages 11-13) blastomeres segregate into two populations, viz., 1) a population of deep blastomeres that will disperse as single motile cells, and 2) a hemispherical shell of outer blastomeres that flattens to form an enveloping cell layer (EVL). When epiboly of the EVL and the yolk syncytial layer (YSL) commences (stage 14), deep blastomeres clump together as a consolidation mass and then migrate outward as single cells on the YSL. When epiboly is concluded (stage 19), deep blastomeres have completely dispersed. If diapause does not intervene, the dispersed phase lasts only a few days. Subsequently, the dispersed cells come together to form a definitive aggregate (stage 27). Embryogenesis within the reaggregated mass of previously dispersed cells produces a typical teleost embryo. Early development in Cynolebias resembles that of other South American annual fishes, such as Austrofundulus, in that a phase of deep blastomere dispersion and reaggregation spatially and temporally separates epiboly from embryogenesis. Several features of development markedly differ from Austrofundulus. There are far fewer (250 vs. 2,500) deep blastomeres. Deep cells of Cynolebias are flattened rhomboids with filipodial extensions in contrast to the amoeboid cells of Austrofundulus. Blastomeres of dispersion and reaggregation stages in Cynolebias send out numerous cell surface extensions onto the YSL and in contact with one another, and often line up in rows as do some African annual fishes, e.g., Nothobranchius. During Dispersion II (stage 21), Reaggregation I (stage 22), and Reaggregation II (stage 23), deep cells move in an oriented pattern with respective mean velocities of 3.48 +/- 0.91, 1.28 +/- 0.46, and 1.31 +/- 0.31 microns/minute. Cells move toward a granular mass of unknown composition, located at the YSL-yolk interface in the lower hemisphere of the egg. This mass appears to coincide with the site of cell reaggregation.

The Variable Effects of Ambient and Artificial Light:Dark Cycles On Embryonic Diapause in A Laboratory Population of the Annual Fish Nothobranchius Guentheri

ABSTRACT The effects of light:dark cycles (L;D) on embryonic development in a laboratory population of the East African annual fish Nothobranchius guentheri were studied. Under ambient light conditions (40° N) there was a low frequency of embryos entering diapause between June and October. Beginning in November there was an increasing frequency of diapausing embryos with a peak in December, and a lower frequency by February. Under artificial light conditions there was an increasing frequency of diapausing embryos as the L:D changed from 16:8 to 9:15. When individual fish or groups of fish were followed it was found that, even under the same light conditions, variable frequencies of diapausing and non-diapausing embryos were produced and that the frequencies often changed with time. The L: D cycle under which the embryos were incubated had no effect on diapause. As in some species of insects the ‘diapause factor’ was of maternal origin. The ability of the fish to produce both diapausing and non-diapausing embryos under the same and variable L : D is most likely an adaptive trait related to the survival of the fish in the harsh environments of alternating rainy and dry seasons.

The survival of embryos of the annual fish Nothobranchius guentheri exposed to temperature extremes and the subsequent effects on embryonic Diapause

AbstractNothobranchius guentheri is an East African annual fish found in temporary bodies of water in areas of alternating rainy and dry seasons. During the dry season the embryos may be exposed to temperature extremes. Embryos have been reported to undergo diapause (developmental arrest), and are thus able to withstand the variable durations of the dry seasons. Since the embryos appear to endure long durations in diapause, and since in diapause they may be exposed to large variations in temperature, we have attempted to determine the thermal tolerance and responsiveness of embryos at diapause and at stages exclusive of diapause. Embryos were exposed to temperatures ranging from −8‐40°C for variable durations of time, and survival, abnormalities and developmental rates determined. Both Diapause I and Diapause II embryos were much less sensitive to temperature extremes than were the other stages, and Diapause II was much less sensitive than Diapause I. Abnormalities only occurred under certain temperature exposures but showed the highest frequency at the early stages of development. Short exposure of Diapause I embryos to −8 and 3.4°C eliminated Diapause II, while short exposure to 40°C enhanced its duration. Although it has been proposed that the onset and duration of the diapauses are denendent on the oxygen concentration of the water, we propose that temper ature may also be another imposrant factor.