Molecular phylogeny of Eastern Pacific porcelain crabs, genera Petrolisthes and Pachycheles, based on the mtDNA 16S rDNA sequence: phylogeographic and systematic implications

Porcelain crabs, genera Petrolisthes and Pachycheles, are diverse and abundant members of the eastern Pacific near-shore decapod crustacean community. Morphology-based taxonomic analyses of these crabs have determined groupings of affiliated species, but phylogenetic relationships remain unknown. We used sequence data from the mitochondrial 16S rRNA gene of 46 species of eastern Pacific porcelain crabs to perform phylogenetic analyses by distance and parsimony methods. Our results are used to compare the taxonomic significance of morphological and molecular characters, to examine sequence divergence rates of crab 16S rRNA genes, and to analyze the phylogeographic history of these crabs. Our phylogenetic trees indicate that the genus Petrolisthes is divided into two main clades, reflecting morphological features. One clade contains primarily tropical species, and the other contains species from throughout the eastern Pacific, as well as species in the genera Allopetrolisthes and Liopetrolisthes. Phylogenetic trees of Pachycheles suggest an antitropical distribution; north and south temperate species form one clade and tropical species form a second clade. Sequence divergence rates of the 16S rRNA gene from three pairs of geminate species can be used to date divergence times, and we discuss porcelain crab phylogeographic patterns in relation to paleogeographic events.     

A comparative analysis of the upper thermal tolerance limits of eastern Pacific porcelain crabs, genus Petrolisthes: influences of latitude, vertical zonation, acclimation, and phylogeny

Marine intertidal organisms are subjected to a variety of abiotic stresses, including aerial exposure and wide ranges of temperature. Intertidal species generally have higher thermal tolerance limits than do subtidal species, and tropical species have higher thermal tolerance limits than do temperate species. The adaptive significance of upper thermal tolerance limits of intertidal organisms, however, has not been examined within a comparative context. Here, we present a comparative analysis of the adaptive significance of upper thermal tolerance limits in 20 congeneric species of porcelain crabs, genus Petrolisthes, from intertidal and subtidal habitats throughout the eastern Pacific. Upper thermal tolerance limits are positively correlated with surface water temperatures and with maximal microhabitat temperatures. Analysis of phylogenetically independent contrasts (from a phylogenetic tree on the basis of the 16s rDNA gene sequence) suggests that upper thermal tolerance limits have evolved in response to maximal microhabitat temperatures. Upper thermal tolerance limits increased during thermal acclimation at elevated temperatures, the amount of increase being greater for subtidal than for intertidal species. This result suggests that the upper thermal tolerance limits of some intertidal species may be near current habitat temperature maxima, and global warming thus may affect the distribution limits of intertidal species to a greater extent than for subtidal species.     

Hair-trigger autotomy in porcelain crabs is a highly effective escape strategy

The benefits of autotomy, the voluntary shedding of limbs, havebeen adequately demonstrated in vertebrates but are poorlystudied in invertebrates. We provide strong experimental evidencefor an antipredatory benefit to autotomy in two porcelain crabs(Petrolisthes cinctipes and P. manimaculis). Since the outcomeof autotomy depends critically on the species of predator andprey involved, we first surveyed field populations of porcelaincrabs to identify ecologically relevant predators to use insubsequent experiments. We then examined the escape tacticsof the porcelain crabs in response to the four potential predatorswe identified, all larger predatory crabs. Cheliped autotomywas induced by three of the predator species (Cancer antennarius,Hemigrapsus nudus, Pachygrapsus crassipes); the fourth didnot attack porcelain crabs. Autotomy occurred in response to34% of all attacks, and in 67% of attacks in which the preywas held at some point by only the cheliped. Autotomy was ahighly effective escape mechanism against these predators;58 of 59 crabs that autotomized escaped, usually while thepredator stopped pursuit to eat the shed cheliped. Relianceon autotomy as a primary mechanism for escaping predators maybe particularly common in small crabs that cannot adequatelydefend themselves by other means and in suspension-feedingcrabs that do not need their chelipeds for feeding.     

Identification of proteins interacting with lactate dehydrogenase in claw muscle of the porcelain crab Petrolisthes cinctipes

Biochemical adaptation of enzymes involves conservation of activity, stability and affinity across a wide range of intracellular and environmental conditions. Enzyme adaptation by alteration of primary structure is well known, but the roles of protein–protein interactions in enzyme adaptation are less well understood. Interspecific differences in thermal stability of lactate dehydrogenase (LDH) in porcelain crabs (genus Petrolisthes) are related to intrinsic differences among LDH molecules and by interactions with other stabilizing proteins. Here, we identified proteins that interact with LDH in porcelain crab claw muscle tissue using co-immunoprecipitation, and showed LDH exists in high molecular weight complexes using size exclusion chromatography and Western blot analyses. Co-immunoprecipitated proteins were separated using 2D SDS PAGE and analyzed by LC/ESI using peptide MS/MS. Peptide MS/MS ions were compared to an EST database for Petrolisthes cinctipes to identify proteins. Identified proteins included cytoskeletal elements, glycolytic enzymes, a phosphagen kinase, and the respiratory protein hemocyanin. Our results support the hypothesis that LDH interacts with glycolytic enzymes in a metabolon structured by cytoskeletal elements that may also include the enzyme for transfer of the adenylate charge in glycolytically produced ATP. Those interactions may play specific roles in biochemical adaptation of glycolytic enzymes.     

Testing the abundant‐centre hypothesis using intertidal porcelain crabs along the Chilean coast: linking abundance and life‐history variation

Aim The abundant‐centre hypothesis (ACH) is based on the assumption that physiological constraints limit populations at the edges of their distributional range, yet the geographical variation of physiological performance or life‐history traits has rarely been examined. Here we examine the applicability of the ACH in a marine system by testing whether physiological predictions are reflected in large‐scale variations of life‐history traits. Location The Chilean coast (18°–42° S), encompassing more than 2500 km along the Pacific coast of South America. Methods Five porcelain crab species (Petrolisthes granulosus, Petrolisthes laevigatus, Petrolisthes tuberculatus, Petrolisthes violaceus and Allopetrolisthes angulosus) were sampled on intertidal boulder beaches at 13 sampling sites. For each species and site we evaluated: (1) relative abundance (density), (2) maximum size, (3) size at maturity, (4) sex ratio, (5) proportion of ovigerous females, and (6) presence of recruits. The shape of the spatial distribution of each trait was evaluated statistically against the prediction of four hypothetical models (normal, ramped‐south, ramped‐north and abundant‐edge). Results The relative abundance and life‐history traits showed different spatial patterns among species. Relative abundance (across sites) was fitted by a normal model in only two species. No model fitted the spatial variation in body size and size at first maturity, which showed a slight but monotonic poleward increase in all species. Sex ratio showed a prominent hump‐shaped pattern, with females prevailing in the centre of the ranges and males dominating towards the range boundaries; this pattern was statistically significant in three of the five studied species. The proportion of ovigerous females showed no clear latitudinal trends, and mature individuals were observed across most of the geographical range of the species. However, recruits tended to be absent towards the southern (poleward) boundaries of the distribution. Main conclusions The ACH does not apply to all species equally. The link between abundance and life‐history traits is complex and variable among the porcelain crab species studied. Overall, the observed patterns were consistent with the idea that equatorward boundaries might be controlled by physiological restrictions mainly affecting adult survival, whereas poleward boundaries might be shaped by limitations in reproductive output and larval survival. Our results underline the importance of incorporating ecological, physiological and life‐history studies in future tests of the ACH.     

马鞍列岛海洋牧场褐菖鲉的摄食习性

海洋牧场是近岸渔业资源保护和恢复的有效途径,为评估海洋牧场建设对岩礁性鱼类的影响,本文基于马鞍列岛海洋牧场建设前后资源调查数据,利用胃含物分析和稳定同位素测试,研究了褐菖鲉的摄食习性.结果表明:海洋牧场建设前褐菖鲉主要摄食端足类、蟹类和头足类,优势饵料生物种类为麦秆虫、日本枪乌贼、日本蟳、日本岩瓷蟹和滩栖阳遂足等.随体长的增大,蟹类的比例增加,而端足类的比例减少.洋牧场建设后褐菖鲉主要摄食蟹类、端足类和鱼类,优势饵料生物种类为双斑蟳、日本蟳、麦秆虫和赤鼻棱鳀等.随体长的增大,鱼类和蟹类的比例增加,而端足类的比例先增加后减少.根据稳定同位素分析结果,海洋牧场区褐菖鲉可以划分为体长<10.0 cm、10.0～13.9.0 cm和≥14.0 cm 3个摄食群体,平均营养级为3.40级.     

Bimodal respiration, water balance and acid-base regulation in a high-shore crab, Cyclograpsus lavauxi H. Milne Edwards

Cyclograpsus lavauxi H. Milne Edwards, 1853 occurs under boulders near the littoral fringe, where it is wetted only briefly, if at all, in each tide. Mean water content of sea-water equilibrated crabs was 62.3 % of body weight. On warm, windy days crabs on the shore lose > 17.5% of body water. Water loss for crabs of 1.6 g mean body weight was 0.29 % water content · h ⁻¹ · mm Hg saturation deficit ⁻¹, which is similar to that of species occupying positions lower on the shore. C. lavauxi is, however, able to tolerate comparatively greater losses of body water (up to 36%) compared with species from lower shore levels.

As the gills dry, the lamellae separate into regular clumps which may help to maintain gas exchange. Resting oxygen consumption (V_o2) after 3–5 h settlement was similar in water and air. During enforced activity, V_o2 increased by factors of 5.3 in water and 2.6 in air at a standardized body weight of 1.5 g, indicating an appreciable aerobic scope in both media. Loss of up to 16% of body water did not depress resting aerial V_o2 or aerobic scope.

In resting crabs there was no change in haemolymph pH after 24 h of either immersion or emersion, but haemolymph [Ca²⁺], [HCO⁻₃] + [CO²⁻₃], and calculated P_co2 all rose in air relative to aquatic values. These results suggest that cuticular or other endogenous CaCO₃ is mobilized to compensate the respiratory acidosis in air. The implications of such physiological properties for aquatic and aerial activity on the shore are discussed.     

Metabolic rate and membrane fatty acid composition in birds: a comparison between long-living parrots and short-living fowl

Both basal metabolic rate (BMR) and maximum lifespan potential (MLSP) vary with body size in mammals and birds and it has been suggested that these are mediated through size-related variation in membrane fatty acid composition. Whereas the physical properties of membrane fatty acids affect the activity of membrane proteins and, indirectly, an animal’s BMR, it is the susceptibility of those fatty acids to peroxidation which influence MLSP. Although there is a correlation between body size and MLSP, there is considerable MLSP variation independent of body size. For example, among bird families, Galliformes (fowl) are relatively short-living and Psittaciformes (parrots) are unusually long-living, with some parrot species reaching maximum lifespans of more than 100 years. We determined BMR and tissue phospholipid fatty acid composition in seven tissues from three species of parrots with an average MLSP of 27 years and from two species of quails with an average MLSP of 5.5 years. We also characterised mitochondrial phospholipids in two of these tissues. Neither BMR nor membrane susceptibility to peroxidation corresponded with differences in MLSP among the birds we measured. We did find that (1) all birds had lower n-3 polyunsaturated fatty acid content in mitochondrial membranes compared to those of the corresponding tissue, and that (2) irrespective of reliance on flight for locomotion, both pectoral and leg muscle had an almost identical membrane fatty acid composition in all birds.     

Performance of ground-based and aerial survey methods for monitoring wildlife assemblages in a conservation area of northern Tanzania

Validating and improving field-sampling techniques for estimating wildlife community composition and population size is essential for wildlife management and conservation. We conducted ground distance sampling surveys along line transects and block counts from a small aircraft in Manyara Ranch in Northern Tanzania and contrasted estimates of species richness and species-specific densities from both sampling techniques. We used regression analyses (logistic regression and generalized linear mixed models) and model selection to investigate whether a species’ body size, group size, body color, as well as vegetation cover explained the variation in species presence/absence and relative density differences in aerial vs. ground-based sampling. Ground surveys detected significantly more species than aerial surveys. However, aerial surveys detected three species that were missed by ground surveys (African lions, African buffalo, and spotted hyena). Model selection suggested that species with smaller body mass and small group sizes were more likely to be missed in aerial surveys. Densities estimated from the aerial surveys were generally but non-significantly lower than the densities estimated from the ground surveys, with the exception of density estimates for African elephants which were slightly higher from aerial surveys. Density differences between the two methods were greater for species with small group size, light body color, and in areas with denser vegetation cover; these variables explained 75% of the variation in density differences between the two survey methods. Albeit being similar in operational costs in our relatively small study area, ground surveys yielded (1) more complete information with respect to wildlife community composition and (2) density estimates were mostly higher and (3) more precise and (4) appear more feasible to be implemented in community-based conservation schemes.     

Leg length and temperature determine the use of unipedal roosting in birds

The function of standing on one leg in birds has long been attributed to reducing heat loss from the unfeathered legs to the external environment. Whilst a handful of single‐species studies correlate the use of the behaviour with ambient temperature, the degree to which it is used across taxa is unknown. Given that leg‐length varies between species, the length of the leg (relative to body size) may mediate the use of this thermoregulatory behaviour, such that birds with longer legs should roost on one leg more than those with relatively shorter legs at any given ambient temperature. We tested this prediction through field observations and comparative analyses of nine shorebird species, with varying tarsi length relative to body size. Six of the nine species examined used unipedal standing more as temperatures decrease, indicating its role as a heat conservation behaviour. We also found that species with relatively longer legs roosted on one leg more frequently across a wide range of temperatures. Species with shorter leg lengths likely rely less on this posture to insulate the relatively smaller surface area of the legs. Our findings showed that the long accepted notion that birds stand on one leg more at colder temperatures holds, and that species with smaller relative leg length were less reliant on this behaviour to minimise heat loss from these bare appendages.     

Spacing and movement in the green porcelain crab Petrolisthes armatus: evidence for male competition and mate guarding

The green porcelain crab, Petrolisthes armatus, is a common invasive species on inter-tidal oyster reefs in the South Atlantic Bight whose behavior is largely unknown. We assessed the effects of the presence of opposite-sex conspecifics on adult crab spacing at a low density to infer potential mate acquisition behaviors. Adult crabs held in all-male, all-female, and mixed-sex treatments were observed twice daily over five days to determine nearest-neighbor (NN) distances and relative activity. Males in the mixed-sex treatment with the shortest NN distances (NN₁) were most commonly (1) closest to females, and, when compared to NN₁ males in the all-male treatment, were (2) closer to their NN and (3) had lower activity levels. Female movement did not differ between treatments. Percent ovigerous females at the end of trials were higher in the mixed-sex treatment, reflecting mating activity, which when observed, involved inter-molt females. Mate guarding, rather than pure-searching behavior, was the mating behavior most consistent with observed spacing and movement patterns.     

Sexual dimorphism in leg length among orb-weaving spiders: a possible role for sexual cannibalism

The degree and direction of sexual dimorphism across different species is commonly attributed to differences in the selection pressures acting on males and females. The extent of these differences is especially apparent in species that practise sexual cannibalism, where the female attempts to capture and eat a courting male. Here, we investigate the relationship between sexual dimorphism in size and leg length, sexual cannibalism and courtship behaviour in three taxonomic groups of orb-weaving spiders, using morphological data from 249 species in 36 genera. Females are larger than males in all three taxonomic groups, and males have relatively longer legs than females in both the Araneinae and Tetragnathidae. Across genera within each taxonomic group, male body size is positively correlated with both female body size and male leg length, and female body size is positively correlated with female leg length. Sexual size dimorphism is negatively correlated with relative male leg length within the Araneinae, but not within either the Tetragnathidae or the Gasteracanthinae. There was no negative correlation between sexual size dimorphism and relative female leg length in any taxonomic group. We argue that the relationship between sexual size dimorphism and relative male leg length within the Araneinae may be the result of selection imposed by sexual cannibalism by females.     

Constraint on Reproductive Output in Brachyuran Crabs: Pinnotherids Test the Rule

Brood weight and number of eggs per brood are primarily determinedby female body size in brachyuran crabs. Brood weight exhibitsan isometric relationship spanning more than four orders ofmagnitude in body weight among 33 non-pinnotherid species, withbrood size being constrained to about 10% of female body weightby the space available for yolk accumulation within the cephalothorax.In contrast, two pinnotherids (Pinnotheres ostreum, Fabiasubquadrata)have relative brood sizes which are 66% and 97%, respectively,of body weight. The allometric constraint is circumvented inthese extraordinarily large brood sizes by two pinnotherid featureswhich allow more space for yolk accumulation. Unlike other brachyurans,their ovaries extend out of the cephalothorax into the abdomen,and calcification of their exoskeleton is greatly reduced, potentiallyallowing distension of the body. As in crabs, pinnotherid eggsize is highly variable among species, which tends to increasethe variance among species in fecundity per brood, making pinnotheridfecundities not extraordinary. Thenumber of broods producedper year in these two species is limited by short reproductiveseasons, which tends to bring their annual reproductive outputin line with other brachyurans. The trade-offs among reproductivevariables in pinnotherids is adaptive for the small body sizeand parasitic niche of these species.     

Muscle mitochondrial capacity exceeds maximal oxygen delivery in humans

Across a wide range of species and body mass a close matching exists between maximal conductive oxygen delivery and mitochondrial respiratory rate. In this study we investigated in humans how closely in-vivo maximal oxygen consumption (VO(2) max) is matched to state 3 muscle mitochondrial respiration. High resolution respirometry was used to quantify mitochondrial respiration from the biopsies of arm and leg muscles while in-vivo arm and leg VO(2) were determined by the Fick method during leg cycling and arm cranking. We hypothesized that muscle mitochondrial respiratory rate exceeds that of systemic oxygen delivery. The state 3 mitochondrial respiration of the deltoid muscle (4.3±0.4 mmol o(2)kg(-1) min(-1)) was similar to the in-vivo VO(2) during maximal arm cranking (4.7±0.5 mmol O(2) kg(-1) min(-1)) with 6 kg muscle. In contrast, the mitochondrial state 3 of the quadriceps was 6.9±0.5 mmol O(2) kg(-1) min(-1), exceeding the in-vivo leg VO(2) max (5.0±0.2 mmol O(2) kg(-1) min(-1)) during leg cycling with 20 kg muscle (P<0.05). Thus, when half or more of the body muscle mass is engaged during exercise, muscle mitochondrial respiratory capacity surpasses in-vivo VO(2) max. The findings reveal an excess capacity of muscle mitochondrial respiratory rate over O(2) delivery by the circulation in the cascade defining maximal oxidative rate in humans.     

Testing the role of male–male competition in the evolution of sexual dimorphism: a comparison between two species of porcelain crabs

Theory predicts marked sexual dimorphism in terms of body size and body structures used as weapons (e.g. chelipeds) in gonochoric species with intense male sexual competition for receptive females and reduced or no sexual dimorphism in species where competition among males is trivial. We tested this hypothesis using a pair of closely‐related species of symbiotic porcelain crabs as a model. In one species that inhabits sea anemones solitarily, competition among males for receptive females is unimportant. In a second species that dwells as dense aggregations on sea urchins, male–male competition for sexual partners is recurrent. We expected considerable sexual dimorphism in body size and weaponry in the urchin‐dwelling crab and reduced sexual dimorphism in the anemone‐dwelling crab. In agreement with expectations, in the urchin‐dwelling crab, male body size was, on average, larger than that of females and males invested considerably more to cheliped length than females. Also supporting theoretical considerations, in the anemone‐dwelling crab, sexual dimorphism in terms of body size was not detected and differences between the sexes in investment to cheliped length were minor. Interestingly, chelipeds were more developed both in males and females of the anemone‐dwelling crab than in the urchin‐dwelling crab as a result of the importance of these structures for monopolization of their naturally scarce anemone hosts. Another difference between the studied species was the existence of two clearly distinguishable ontogenetic phases in males of the urchin‐dwelling crab but not in males of the anemone‐dwelling crab. Whether the two different male morphs display different male reproductive strategies in the urchin‐dwelling crab remains to be addressed. Other conditions that might additionally explain the observed differences in sexual dimorphism (e.g. female mate choice) between the studied species remain to be explored. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 548–558.     

The impact of limb autotomy on mate competition in blue crabs Callinectes sapidus Rathbun

Summary This study is the first to demonstrate experimentally that autotomy (self-amputation of a body part) adversely affects competition for mates. Experiments were conducted using blue crabs Callinectes sapidus Rathbun to examine the consequences of limb loss and pairing precedence on mate acquisition by males. Two adult males of equivalent size were introduced sequentially into pools containing a sexually-receptive female and observed after 24 h and 48 h. One male in each pair was left intact, while the other experienced: (1) no autotomy, (2) autotomy of one cheliped, or (3) autotomy of both chelipeds, one walking leg, and one swimming leg. In the absence of a competitor (first 24 h), both intact and injured males established precopulatory embraces with females. Intact males were highly successful (84–95%) in defending females from intact or injured intruders in the second 24 h period. Both autotomy treatments, however, significantly reduced the ability of males to defend females from intact intruders. Females in experiments suffered greater frequency of limb loss than did males. In the field, paired blue crabs showed significantly higher incidence of limb loss than unpaired crabs. Limb loss frequency increases with body size, and field observations indicated that larger males may be more successful than smaller males in obtaining females. Both experimental manipulations and field studies provide strong evidence for mate competition in this ecologically and commercially important portunid species.     

The constant proportion of grana and stroma lamellae in plant chloroplasts

The relative proportion of stroma lamellae and grana end membranes was determined from electron micrographs of 58 chloroplasts from 21 different plant species. The percentage of grana end membranes varied between 1 and 21% of the total thylakoid membrane indicating a large variation in the size of grana stacks. By contrast the stroma lamellae account for 20.3 ± 2.5 ( sd )% of the total thylakoid membrane. A plot of percentage stroma lamellae against percentage of grana end membranes fits a straight line with a slope of zero showing that the proportion of stroma lamellae is independent of the size of the grana stacks. That stroma lamellae account for about 20% of the thylakoid membrane is in agreement with fragmentation and separation analysis (Gadjieva et al . Biochim. Biophys. Acta 144: 92–100, 1999). Chloroplasts from spinach, grown under high or low light, were fragmented by sonication and separated by countercurrent distribution into two vesicle populations originating from grana and stroma lamellae plus end membranes, respectively. The separation diagrams were very similar lending independent support for the notion that the proportion of stroma lamellae is constant. The results are discussed in relation to the composition and function of the chloroplast in plants grown under different environmental conditions, and in relation to a recent quantitative model for the thylakoid (Albertsson, Trends Plant Sci. 6: 349–354, 2001).