Exosymbiotic microbes within fermented pollen provisions are as important for the development of solitary bees as the pollen itself

1. Developing bees derive significant benefits from the microbes present within their guts and fermenting pollen provisions. External microbial symbionts (exosymbionts) associated with larval diets may be particularly important for solitary bees that suffer reduced fitness when denied microbe‐colonized pollen.
2. To investigate whether this phenomenon is generalizable across foraging strategy, we examined the effects of exosymbiont presence/absence across two solitary bee species, a pollen specialist and generalist. Larvae from each species were reared on either microbe‐rich natural or microbe‐deficient sterilized pollen provisions allocated by a female forager belonging to their own species (conspecific‐sourced pollen) or that of another species (heterospecific‐sourced pollen). Our results reveal that the presence of pollen‐associated microbes was critical for the survival of both the generalist and specialist larvae, regardless of whether the pollen was sourced from a conspecific or heterospecific forager.
3. Given the positive effects of exosymbiotic microbes for larval fitness, we then examined if the magnitude of this benefit varied based on whether the microbes were provisioned by a conspecific forager (the mother bee) or a heterospecific forager. In this second study, generalist larvae were reared only on microbe‐rich pollen provisions, but importantly, the sources (conspecific versus heterospecific) of the microbes and pollen were experimentally manipulated.
4. Bee fitness metrics indicated that microbial and pollen sourcing both had significant impacts on larval performance, and the effect sizes of each were similar. Moreover, the effects of conspecific‐sourced microbes and conspecific‐sourced pollen were strongly positive, while that of heterospecific‐sourced microbes and heterospecific‐sourced pollen, strongly negative.
5. Our findings imply that not only is the presence of exosymbionts critical for both specialist and generalist solitary bees, but more notably, that the composition of the specific microbial community within larval pollen provisions may be as critical for bee development as the composition of the pollen itself.

     

Diverse Microbiota Identified in Whole Intact Nest Chambers of the Red Mason Bee Osmia bicornis (Linnaeus 1758)

Microbial activity is known to have profound impact on bee ecology and physiology, both by beneficial and pathogenic effects. Most information about such associations is available for colony-building organisms, and especially the honey bee. There, active manipulations through worker bees result in a restricted diversity of microbes present within the colony environment. Microbial diversity in solitary bee nests remains unstudied, although their larvae face a very different situation compared with social bees by growing up in isolated compartments. Here, we assessed the microbiota present in nests and pre-adults of Osmia bicornis, the red mason bee, by culture-independent pyrosequencing. We found high bacterial diversity not comparable with honey bee colonies. We identified a variety of bacteria potentially with positive or negative interactions for bee larvae. However, most of the other diverse bacteria present in the nests seem to originate from environmental sources through incorporated nest building material and stored pollen. This diversity of microorganisms may cause severe larval mortality and require specific physiological or symbiotic adaptations against microbial threats. They may however also profit from such a diverse environment through gain of mutualistic partners. We conclude that further studies of microbiota interaction in solitary bees will improve the understanding of fitness components and populations dynamics.     

How caterpillars avoid overheating: behavioral and phenotypic plasticity of pipevine swallowtail larvae

We tested the hypothesis that larvae of the pipevine swallowtail butterfly, Battus philenor, employ behavioral and phenotypic plasticity as thermoregulatory strategies. These larvae are phenotypically varied across their range with predominantly black larvae (southeastern USA and California) and red larvae (western Texas, Arizona) occurring in different regions. Two years of field observations in south Texas indicate that the proportion of red larvae increases with increasing daily temperatures as the growing season progresses. Larvae were also observed to shift their microhabitats by climbing on non-host vegetation and avoided excessive heat in their feeding microhabitat. Larvae of ten half-sib families from populations in south Texas and California, reared under different temperature regimes in common garden experiments, exhibited plasticity in larval phenotype, with larvae from both populations producing the red phenotype at temperatures greater than 30°C and maintaining the black phenotype at cooler temperatures. However, larvae from Texas were more tolerant of higher temperatures, showing no decrease in growth rate in the highest temperature (maximum seasonal temperature) treatment, compared to the California population. In a field experiment, black larvae were found to have higher body temperatures when exposed to sunlight compared to red larvae. These results suggest that microhabitat shifts and the color polyphenism observed in pipevine swallowtail larvae may be the adaptive strategies that enable larvae to avoid critical thermal maximum temperatures.     

No evidence that gut microbiota impose a net cost on their butterfly host

Gut microbes are believed to play a critical role in most animal life, yet fitness effects and cost–benefit trade‐offs incurred by the host are poorly understood. Unlike most hosts studied to date, butterflies largely acquire their nutrients from larval feeding, leaving relatively little opportunity for nutritive contributions by the adult's microbiota. This provides an opportunity to measure whether hosting gut microbiota comes at a net nutritional price. Because host and bacteria may compete for sugars, we hypothesized that gut flora would be nutritionally neutral to adult butterflies with plentiful food, but detrimental to semistarved hosts, especially when at high density. We held field‐caught adult Speyeria mormonia under abundant or restricted food conditions. Because antibiotic treatments did not generate consistent variation in their gut microbiota, we used interindividual variability in bacterial loads and operational taxonomic unit abundances to examine correlations between host fitness and the abdominal microbiota present upon natural death. We detected strikingly few relationships between microbial flora and host fitness. Neither total bacterial load nor the abundances of dominant bacterial taxa were related to butterfly fecundity, egg mass or egg chemical content. Increased abundance of a Commensalibacter species did correlate with longer host life span, while increased abundance of a Rhodococcus species correlated with shorter life span. Contrary to our expectations, these relationships were unchanged by food availability to the host and were unrelated to reproductive output. Our results suggest the butterfly microbiota comprises parasitic, commensal and beneficial taxa that together do not impose a net reproductive cost, even under caloric stress.     

Trabala Vishnou Gigantina Yang (Lepidoptera: Lasiocampidae) Larval Fitness on six Sympatric Plant Species in Sea-Buckthorn Forest

Trabala vishnou gigantina Yang (Lepidoptera: Lasiocampidae) is a major pest that damages the sea-buckthorn, Hippophae rhamnoides. We observed and compared the feeding preferences of T. vishnou gigantina larvae on six sympatric plant species in a two-choice test. We also compared T. vishnou gigantina fitness, as measured by the following variables: larvae weight, developmental period, pupae versus adult weight, longevity, and fecundity rates. Between host and non-host plants, larvae showed a strong preference for their natural host (sea-buckthorn), followed by apricot, poplar, and willow. Caragana and locust were the least preferred plants when the natural host plant was not present. Larvae reared on sea-buckthorn possessed greater pre-pupal weight, had lower mortality, and developed more quickly into heavier pupae than either poplar-raised or willow-raised larvae. Fecundity was highest on sea-buckthorn, second highest on apricot, and lowest on poplar. Longevity (of both females and males) was not significantly different across plant species. These results clearly demonstrate that T. vishnou gigantina larvae are able to distinguish between host versus non-host plants, and that their preference translates to increased fitness. Possible, non-mutually exclusive explanations for observed preference and fitness differences include variation in required nutritional content across plant species or the presence of plant traits (morphological features or chemical metabolites) that negatively affect larval development. While the exact mechanisms are unknown, these data may be useful for the development of appropriate counter-measures to the damage caused by T. vishnou gigantina on sea-buckthorn.     

Palaeocene origin of the Neotropical lineage of cleptoparasitic bees Ericrocidini‐Rhathymini (Hymenoptera,Apidae)

Cleptoparasitic bees abandoned pollen‐collecting for their offspring and lay their eggs in other bees' provisioned nests. Also known as cuckoo bees, they belong to several lineages, but are especially diverse in Apinae. We focused on a lineage of apine cleptoparasitic bees, the clade Ericrocidini + Rhathymini, which attack nests of oil‐collecting bees. We sequenced five genes for 20 species in all genera of this clade plus a large outgroup to reconstruct the phylogeny and estimate divergence times. We confirmed the monophyly of the clade Ericrocidini + Rhathymini and its position inside the ericrocidine line together with the tribes Protepeolini, Isepeolini, Osirini and Coelioxoidini. Our results corroborated the current taxonomic classification. Ericrocis is sister to all Ericrocidini and the position of Acanthopus and the most diverse genus Mesoplia were inconclusive. Ericrocidini + Rhathymini diverged from other apine cleptoparasitic lineages 74 Ma in the Cretaceous and from each other in the Palaeocene at 61 Ma. Considering the robust molecular evidence of their sister relationships, the striking differences in the morphology of first‐instar larvae of the two groups may represent adaptations to the nesting biology of their hosts. As other cleptoparasites in the ericrocidine line, Ericrocidini and Rhathymini possess larvae which are adapted to kill the immature host and to feed on floral oil provided by the host female. The evolution of host specialization in the line Ericrocidini + Rhathymini retroceded to the Eocene when they differentiated synchronously with their hosts, Centris and Epicharis .     

Microbial Community Profiling to Investigate Transmission of Bacteria Between Life Stages of the Wood-Boring Beetle, Anoplophora glabripennis

Many insects harbor specific bacteria in their digestive tract, and these gut microbiota often play important roles in digestion and nutrient provisioning. While it is common for a given insect species to harbor a representative gut microbial community as a population, how this community is acquired and maintained from generation to generation is not known for most xylophagous insects, except termites. In this study, we examined acquisition of gut microbiota by the wood-feeding beetle, Anoplophora glabripennis, by identifying and comparing microbial community members among different life stages of the insect and with microbes it encounters in the environment. Automated ribosomal intergenic spacer analysis was employed to compare bacterial communities present in the egg and larval stages of A. glabripennis as well as with microbes found in the oviposition site and the surrounding woody tissue. Multivariate analyses were used to identify relationships between sample type and specific bacterial types (operational taxonomic units). From this analysis, bacteria that were derived from the environment, the oviposition site, and/or the egg were identified and compared with taxa found in larvae. Results showed that while some larval microbes were derived from environmental sources, other members of the larval microbial community appear to be vertically transmitted. These findings could lead to a better understanding of which microbial species are critical for the survival of this insect and to development of techniques that could be used to alter this community to disrupt the digestive physiology of the host insect as a biological control measure.     

Use of novel pollen species by specialist and generalist solitary bees (Hymenoptera: Megachilidae)

If trade-offs between flexibility to use a range of host species and efficiency on a limited set underlie the evolution of diet breadth, one resulting prediction is that specialists ought to be more restricted than generalists in their ability to use novel resource species. I used foraging tests and feeding trials to compare the ability of a generalist and a specialist solitary mason bee species to collect and develop on two pollen species that are not normally used in natural populations (novel pollens). Osmia lignaria (Hymenoptera: Megachilidae) is a generalist pollen feeder; O. californica, is more specialized. Adults of the specialist were more limited in use of novel hosts, but only in some contexts. Both bee species refused to collect one novel pollen. The specialist accepted a second novel pollen only when it was presented along with its normal pollen, whereas the generalist collected novel pollen whether presented alone or with normal pollen. Surprisingly, larvae of the specialist were more flexible than were generalists. The specialist grew well on mixtures of normal and novel pollen species, in some cases better than on its normal host alone. Larvae of the generalist grew more poorly on all diets containing novel pollens than on their normal host. Data on these two species of bees suggest that specialization by itself need not reduce flexibility on novel hosts. The findings also provide information about mechanisms of specialization in bees. Similar to some folivores, specific cues of the pollen host and the bee's interpretation of these contribute, along with foraging economics, to pollen choice by adults. The ability of the larvae to cope with specific components of one pollen species need not interfere with its ability to use others.     

Biotic Resource Needs of Specialist Orchid Pollinators

Orchid pollinators have highly varied life histories with complex biotic resource requirements, about which we have limited knowledge. Among the specialist orchid pollinators are insect predators and parasitoids with specific prey types such as aphids and subterranean scarab larvae; oligolectic bees that collect pollen from limited sources such as bellflowers; euglossine bees and butterflies that collect particular plant chemicals for reproduction and self defense; oil-collecting bees that provision their brood cells with floral oils from a few plants; bees that collect rare floral resins to construct their nest and brood cells; moths and butterflies that require specific larval host plants; mosquitos and horse flies that need blood; and fungus gnats and carrion flies tied to fungi and dead animals. Loss of critical biotic resources and relationships can reduce the abundance of orchid pollinators and/or their effectiveness. Protection of large, plant rich, pesticide-free orchid habitats is key to conserving essential pollinator resources.     

Microbiology of feces of the larval honey bee,Apis mellifera

Methods for collection and microbiological examination of feces of larval honey bees, Apis mellifera, are described. Feces collected on sterile agar were inoculated onto selective media, some of which were acidified to approximate more closely the pH of larval food and the larval gut. A total of 104 microbial isolates were obtained from fecal collections of 20 larvae, although the feces of 4 of these larvae contained no detectable microbes. Microorganisms isolated in order of frequency were Bacillus spp., Gram-variable pleomorphic bacteria (Achromobacter eurydice?), molds (primarily Penicillia), actinomycetes, Gram-negative bacterial rods, and yeasts. It appears that larvae can become inoculated with microorganisms which are found in adult bees and pollen from ingestion of contaminated food. However, evidence for a constant symbiotic microflora which could contribute significant amounts of biochemicals to larvae is lacking.     

洞察景观环境影响蜜蜂之新视角: 肠道微生物

作为生态服务提供者的传粉蜜蜂与景观生态息息相关, 而以农田为主的景观组成显著降低了传粉蜜蜂的多样性。目前调查研究显示, 农田的扩张与蜜蜂多样性下降相关, 且农药残留对蜜蜂损害严重。景观中的开花植物决定了蜜蜂的食物(营养)组成, 其中花粉蛋白含量与蜜蜂的生长发育紧密相关。尽管研究已证实景观环境会显著影响蜜蜂蜂群的发展和个体的生长繁殖能力, 但未来还需要加强景观组成变化直接作用于蜜蜂的机制研究。另一方面, 大量研究表明蜜蜂肠道共生菌是影响宿主健康的重要因素: 可促进宿主吸收营养和抵抗病原菌。作为传粉者, 蜜蜂接触到的主要外部环境——花粉和花蜜都含有特殊的微生物, 很多研究暗示花源微生物是蜜蜂肠道菌来源之一。研究表明景观环境相关的食物(营养)、农药残留以及环境微生物都会显著影响肠道微生物。现有少量的研究证明不同景观的蜜蜂肠道微生物有差异, 景观环境可能通过作用于蜜蜂肠道微生物进而影响蜜蜂健康。然而不同景观环境中的微生物, 尤其是花源微生物和蜜蜂肠道菌之间的关联有待证明。景观对蜜蜂肠道微生物的影响值得研究, 希望可以从肠道菌的视角鉴别对蜜蜂友好的景观环境, 进而指导土地合理利用和蜜蜂保护。     

Inspection and feeding of larvae by worker honey bees (Hymenoptera: Apidae): Effect of starvation and food quantity

Honey bee larvae are frequently inspected and, sometimes, provided with food by adult workers, but the stimuli that elicit the important task of food provisioning have never been investigated. Larvae with their food experimentally deprived received more frequent inspection and feeding visits from nurse bees than normally fed larvae, suggesting that there could be a hunger signal. Food-deprived larvae with artificially supplied larval food received the same rate of feeding visits from nurse bees as did normally fed larvae but still received more inspection visits. These results suggest that stimuli eliciting feeding are different from those for inspection. They also support the hypothesis that worker bees deposit food in a larval cell only when the quantity of food is below a certain minimum threshold that is perceived during larval inspections. A model is presented regarding the stimuli from larvae that result in worker feeding behavior.     

How Does Pollen Chemistry Impact Development and Feeding Behaviour of Polylectic Bees?

Larvae and imagos of bees rely exclusively on floral rewards as a food source but host-plant range can vary greatly among bee species. While oligolectic species forage on pollen from a single family of host plants, polylectic bees, such as bumblebees, collect pollen from many families of plants. These polylectic species contend with interspecific variability in essential nutrients of their host-plants but we have only a limited understanding of the way in which chemicals and chemical combinations influence bee development and feeding behaviour. In this paper, we investigated five different pollen diets (Calluna vulgaris, Cistus sp., Cytisus scoparius, Salix caprea and Sorbus aucuparia) to determine how their chemical content affected bumblebee colony development and pollen/syrup collection. Three compounds were used to characterise pollen content: polypeptides, amino acids and sterols. Several parameters were used to determine the impact of diet on micro-colonies: (i) Number and weight of larvae (total and mean weight of larvae), (ii) weight of pollen collected, (iii) pollen efficacy (total weight of larvae divided by weight of the pollen collected) and (iv) syrup collection. Our results show that pollen collection is similar regardless of chemical variation in pollen diet while syrup collection is variable. Micro-colonies fed on S. aucuparia and C. scoparius pollen produced larger larvae (i.e. better mates and winter survivors) and fed less on nectar compared to the other diets. Pollen from both of these species contains 24-methylenecholesterol and high concentrations of polypeptides/total amino acids. This pollen nutritional “theme” seems therefore to promote worker reproduction in B. terrestris micro-colonies and could be linked to high fitness for queenright colonies. As workers are able to selectively forage on pollen of high chemical quality, plants may be evolutionarily selected for their pollen content, which might attract and increase the degree of fidelity of generalist pollinators, such as bumblebees.     

Factors influencing host choice of the honey bee parasite Varroa jacobsoni Oud

Reproducing Varroa jacobsoni obtained from brood cells of Apis mellifera L. with 13–16 day old bees (pupae) and Varroa mites kept on adult bees for at least 8 days were simultaneously tested for their choice in three host types. Comparisons were made of attractiveness of Varroa jacobsoni to nurse bees, pollen foragers as to larvae from nearly capped brood cells. Host choices were observed in Petri dishes and in an Y-shaped olfactometer. Varroa jacobsoni obtained from capped brood cells showed a stronger preference for nurse bees in Petri dish simultaneous choice tests with pollen foragers or larvae than did mites which were previously kept on adult bees. In olfactometer simultaneous choice tests, the two mite test groups showed no clear difference in preferences for bees of different ages. The preference of Varroa jacobsoni for bees of different ages is therefore not only influenced by host factors but also by intrinsic factors in female mites that depend on the mite's reproductive stage.     

Proteomic and metabolomic analysis reveals rapid and extensive nicotine detoxification ability in honey bee larvae

Despite potential links between pesticides and bee declines, toxicology information on honey bee larvae (Apis mellifera) is scarce and detoxification mechanisms in this development stage are virtually unknown. Larvae are exposed to natural and synthetic toxins present in pollen and nectar through consumption of brood food. Due to the characteristic intensive brood care displayed by honey bees, which includes progressive feeding throughout larval development, it is generally assumed that larvae rely on adults to detoxify for them and exhibit a diminished detoxification ability. We found the opposite. We examined the proteomic and metabolomic responses of in vitro reared larvae fed nicotine (an alkaloid found in nectar and pollen) to understand how larvae cope on a metabolic level with dietary toxins. Larvae were able to effectively detoxify nicotine through an inducible detoxification mechanism. A coordinated stress response complemented the detoxification processes, and we detected significant enrichment of proteins functioning in energy and carbohydrate metabolism, as well as in development pathways, suggesting that nicotine may promote larval growth. Further exploration of the metabolic fate of nicotine using targeted mass spectrometry analysis demonstrated that, as in adult bees, formation of 4-hydroxy-4-(3-pyridyl) butanoic acid, the result of 2′C-oxidation of nicotine, is quantitatively the most significant pathway of nicotine metabolism. We provide conclusive evidence that larvae are capable of effectively catabolising a dietary toxin, suggesting that increased larval sensitivity to specific toxins is not due to diminished detoxification abilities. These findings broaden the current understanding of detoxification biochemistry at different organizational levels in the colony, bringing us closer to understanding the capacity of the colony as a superorganism to tolerate and resist toxic compounds, including pesticides, in the environment.     

An interaction between host and microbe genotypes determines colonization success of a key bumble bee gut microbiota member

There has been a proliferation of studies demonstrating an organism's health is influenced by its microbiota. However, factors influencing beneficial microbe colonization and the evolution of these relationships remain understudied relative to host–pathogen interactions. Vertically transmitted beneficial microbes are predicted to show high levels of specificity in colonization, including genotype matching, which may transpire through coevolution. We investigate how host and bacterial genotypes influence colonization of a core coevolved microbiota member in bumble bees. The hindgut colonizing Snodgrassella alvi confers direct benefits, but, as an early colonizer, also facilitates the further development of a healthy microbiota. Due to predominantly vertical transmission promoting tight evolution between colonization factors of bacteria and host lineages, we predict that genotype‐by‐genotype interactions will determine successful colonization. Germ‐free adult bees from seven bumble bee colonies (host genotypic units) were inoculated with one of six genetically distinct strains of S. alvi. Subsequent colonization within host and microbe genotypes combinations ranged from 0 to 100%, and an interaction between host and microbe genotypes determined colonization success. This novel finding of a genotype‐by‐genotype interaction determining colonization in an animal host‐beneficial microbe system has implications for the ecological and evolutionary dynamics of host and microbe, including associated host‐fitness benefits.     

Potential effects of oilseed rape expressing oryzacystatin-1 (OC-1) and of purified insecticidal proteins on larvae of the solitary bee Osmia bicornis

Despite their importance as pollinators in crops and wild plants, solitary bees have not previously been included in non-target testing of insect-resistant transgenic crop plants. Larvae of many solitary bees feed almost exclusively on pollen and thus could be highly exposed to transgene products expressed in the pollen. The potential effects of pollen from oilseed rape expressing the cysteine protease inhibitor oryzacystatin-1 (OC-1) were investigated on larvae of the solitary bee Osmia bicornis (= O. rufa). Furthermore, recombinant OC-1 (rOC-1), the Bt toxin Cry1Ab and the snowdrop lectin Galanthus nivalis agglutinin (GNA) were evaluated for effects on the life history parameters of this important pollinator. Pollen provisions from transgenic OC-1 oilseed rape did not affect overall development. Similarly, high doses of rOC-1 and Cry1Ab as well as a low dose of GNA failed to cause any significant effects. However, a high dose of GNA (0.1%) in the larval diet resulted in significantly increased development time and reduced efficiency in conversion of pollen food into larval body weight. Our results suggest that OC-1 and Cry1Ab expressing transgenic crops would pose a negligible risk for O. bicornis larvae, whereas GNA expressing plants could cause detrimental effects, but only if bees were exposed to high levels of the protein. The described bioassay with bee brood is not only suitable for early tier non-target tests of transgenic plants, but also has broader applicability to other crop protection products.     

Predation risk assessment by larval reef fishes during settlement-site selection

Predation rates of marine species are often highest during the transition from the pelagic to the benthic life stage. Consequently, the ability to assess predation risk when selecting a settlement site can be critical to survival. In this study, pairwise choice trials were used to determine whether larvae of three species of anemonefish (Amphiprion melanopus, A. percula and Premnas biaculeatus) are able to (1) assess the predation risk of potential anemone settlement sites through olfactory cues alone and (2) alter their settlement choices depending on the options available (host or non-host anemone). When predation risk was assessed with host and non-host anemone species independently, all species of anemonefish significantly chose the odor associated with the low-risk settlement option over the high-risk site. Most importantly, all species of anemonefish selected water with olfactory cues from their host anemone regardless of predation risk when paired against non-host anemone odor. These results demonstrate that larval reef fishes can use olfactory cues for complex risk assessment during settlement-site selection; however, locating the correct habitat is the most important factor when selecting a settlement site.     

The effect of host plant on larval survivorship of the Alaskan swallowtail butterfly (Papilio machaon aliaska)

The Alaskan swallowtail butterfly, Papilio machaon aliaska (Lepidoptera: Papilionidae), uses three plant species as host plants. Cnidium cnidiifolium (Turcz.) Schischk. belongs to the family Apiaceae, the ancestral host‐plant family of the P. machaon group. Artemisia arctica Less. and Petasites frigidus (L.) Franch, in contrast, belong to the distantly related Asteraceae family and were colonized relatively recently by this group of butterflies. Previous work has shown that larval survival is highest on the novel host plants when natural enemies are present in the field. Here I examine whether P. m. aliaska fitness varied when individuals were reared on the three host plants in a common environment, free of environmental and ecological complications such as predation. I collected 12 P. m. aliaska females and measured their reproductive success when their progeny were reared on each of the three host plants. I assessed larval fitness on each of the hosts by recording the percentage pupation and percentage emergence as well as by measuring pupal mass. I found that larvae reared on the ancestral host plant, C. cnidiifolium, had higher fitness than did larvae reared on either of the novel host plants, A. arctica or P. frigidus. Larvae reared on C. cnidiifolium were more likely to pupate, achieved a greater pupal mass, and had higher emergence rates than larvae reared on the novel hosts. I interpret these results to mean that, in the absence of predation, the ancestral host plant is a better host for P. m. aliaska larvae than either novel host and I contend that this result does not appear to support the hypothesis that P. m. aliaska populations in central Alaska are divided into host races.     

Differentiated egg size of the cannibalistic salamander <Emphasis Type="Italic">Hynobius retardatus</Emphasis>

Larvae of the salamander, Hynobius retardatus, are carnivorous, and even though there are two morphs, a typical morph and a broad-headed or “cannibal” morph, both are cannibalistic. They also sometimes eat other large prey, for example larvae of the frog, Rana pirica. In natural habitats, use of both conspecific and R. pirica larvae as food may contribute more strongly to high survival and substantially to fitness when larval densities are higher, because early-stage H. retardatus larvae sometimes experience scarcity of their typical prey. In cannibalistic oviparous amphibians, larger individuals that developed from larger eggs can more efficiently catch and consume larger prey and thus their survival may be better than that of smaller individuals developed from smaller eggs. Populations might therefore diverge in respect of egg size in response to variation in the density of conspecific and R. pirica larvae in natural ponds, with eggs being larger when larval density is higher. I examined how variance in hatchling size correlated with the incidence of cannibalism, and whether increasing larval density in natural ponds correlated with increasing egg size. Variance in initial larval body size facilitated cannibalism, and egg size increased as larval density in the ponds increased. In ponds with high larval density, where cannibalism and large prey consumption is a critical factor in offspring fitness, the production of fewer clutches with larger eggs, and thus of fewer and larger offspring, results in greater maternal fitness. Variation among the mean egg size in populations is likely to represent a shift in optimum egg size across larval density gradients.