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The collagen embedding Sacculina roots is synthesized by hemocytes of Carcinus maenas and is described. Hemocytes engaged in the secretion of collagen as a reaction against a parasite constitute a defensive mechanism still largely unknown within Crustacea.
An overall study of the in vitro plasma coagulation system in the crab Liocarcinus puber has been carried out using various analytical methods, namely thromboelastography, spectrophotometrical examination, and a new one based on changes of the mechanical impedance of the developing clot. From the results reported here the clotting pattern in this species appears surprisingly complex for an invertebrate and unexpectedly closer to that of the vertebrates. Indirect evidences suggest that the fibrinogen polypeptide chains in this species and very likely in the other crustacean, are very different from those of the vertebrates. This would imply that crustacean and vertebrate fibrinogen would have diverged from one another in a far remote past, far beyond the individualization of the vertebrate alpha chain, that is, over 1.5 million years ago.
IgE-antibody reactivity to oysters and crustacea of sera from six oyster-sensitive, seven oyster- and crustacea-sensitive, and 12 crustacea-sensitive subjects was investigated. All six subjects with a history of only oyster sensitivity had minimal RAST reactivity (ratios 2 to 5) to extracts of raw or boiled oysters. Three of the seven oyster- and crustacea-sensitive subjects and six of the 12 crustacea-sensitive, oyster-tolerant or unexposed subjects had elevated RAST ratios to oyster (14 to 41). Generally, elevated oyster RAST correlated with skin prick test reactivity to oyster but not with total serum IgE levels. The oyster RAST values of the 19 crustacea-sensitive subjects (with or without oyster sensitivity) correlated with crustacea RAST reactivity (crab RAST, most significant; shrimp RAST, least significant). Rabbit antisera to crustacea extracts detected precipitating antigens present in extracts of raw or boiled oysters. Significant inhibition of the oyster RAST was obtained with oyster or crustacea extracts. These studies suggest that in the diagnosis of oyster sensitivity the RAST may not be useful and that oyster and crustacea contain common antigenic structures.
Adverse reactions to ingested crustacea are common and may be life-threatening. We studied 14 individuals with histories of such reactions to shrimp by immediate skin tests and RAST with extracts of shrimp, crab, crayfish, and lobster. Nine of these subjects (8/8 atopics and 1/6 nonatopics) had positive immediate skin tests (wheal greater than or equal to 2 mm) and RAST (ratios greater than 3.0) to shrimp. Their skin tests and RAST ratios to the other crustacea were also frequently positive even, in several cases, in the absence of prior exposure. In contrast, only 1/10 volunteers with no history of intolerance to crustacea had a weak positive skin test to raw shrimp. These studies suggest that both skin tests and RAST are useful in the confirmation of hypersensitivity to shrimp in atopic individuals and that cross-reactivity among crustacea may exist.
The sodium pump, or Na(+)+K(+)-ATPase, provides at least part of the driving force for transepithelial movement of monovalent ions across the gills and other transporting tissues in many aquatic animals including the Crustacea. The crustacean Na(+)+K(+)-ATPase, like that in all animal cells, is composed of a catalytic alpha-subunit and an accompanying beta-subunit. The amino acid sequence of the crustacean alpha-subunit is 71-74% identical to vertebrate alpha-subunit sequences. In brachyuran Crustacea, the Na(+)+K(+)-ATPase is more highly expressed in posterior gills compared with anterior and is found predominantly in mitochondria-rich cells that are morphologically and biochemically specialized to mediate NaCl uptake from the medium. When the external salinity is lowered from that of normal seawater, producing conditions in which many euryhaline Crustacea hyperosmo regulate their hemolymph, both the enzymatic activity of the Na(+)+K(+)-ATPase and the gene expression of the alpha-subunit are increased in these tissues. Although the precise regulatory mechanism is not known, evidence suggests that crustacean hyperglycemic hormone may be responsible for the induction of Na(+)+K(+)-ATPase activity. Whether it also plays a role in activation of gene transcription is not known. A comparison of a range of aquatic Crustacea suggests that the level of Na(+)+K(+)-ATPase function in transporting tissues may be correlated with their ability to invade estuarine habitats.
In the discussion on arthropod phylogeny, the structural evolution of compound eyes and optic ganglia in Crustacea and Insecta is an important topic. On the one hand, many morphological features as well as developmental aspects of the visual system in Insecta and Crustacea correspond in so much detail that eye design in these two groups is likely to have a common euarthropodan ancestor. On the other hand, however, some authors advocate a convergent evolution of the crustacean and insect visual system founding their arguments on differences in the arrangement of the visual neuropils and the fibre connections between Malacostraca and Entomostraca (the "entomostracan enigma"). Therefore, information about cellular aspects of visual system formation in entomostracan Crustacea is likely to enliven this debate, but is not yet available. To fill this gap, we examined the proliferation of neuronal stem cells in the developing visual system of the tadpole shrimp Triops longicaudatus (LeConte, 1846) (Entomostraca, Branchiopoda, Phyllopoda, Calmanostraca, Notostraca) by in vivo incorporation of the proliferation marker bromodeoxyuridine and subsequent immunohistochemical detection. Our results indicate that in the developing visual system of T. longicaudatus, three band-shaped zones containing neuronal stem cells are present corresponding to the proliferation zones found in Malacostraca. We therefore conclude that the ontogenetic mechanisms of visual-system formation are evolutionarily conserved (homologous) in Branchiopoda, Malacostraca, and Insecta.
Antigenic and allergenic components in crawfish and lobster extracts were studied using crossed immunoelectrophoretic techniques. Crossed immunoelectrophoresis with rabbit antisera revealed 23 antigens in crawfish and 17 antigens in lobster extracts. Both extracts exhibited structural similarities in antigens mutually and with other crustacea in cross-line immunoelectrophoresis. Crossed radioimmunoelectrophoresis (CRIE) demonstrated 6 crawfish and 4 lobster allergens when individual or pooled sera from radioallergosorbent test (RAST)-positive crustacea-sensitive subjects were used. Since radiostaining was also observed with sera from RAST-negative nonsensitive subjects, specificity of IgE binding was tested using CRIE-inhibition. Preincubation of RAST-positive sera with crawfish or lobster extract decreased radiostaining in CRIE, while no changes occurred when using control sera. These results confirmed the presence of IgE-mediated mechanisms in seafood allergy and demonstrated a number of shared antigenic determinants among crustacea allergens.
The distribution of cholecystokinin-like immunoreactivity was studied in the stomatogastric nervous systems, pericardial organs, and haemolymph of four species of decapod crustacea, by using immunocytochemical and radioimmunoassay techniques. Whereas cholecystokinin-like immunoreactivity was found within the stomatogastric nervous systems of all four species, its distribution in each is unique. Two species (Panulirus interruptus and Homarus americanus) have cholecystokinin-like immunoreactivity within fibers and neuropil of the stomatogastric ganglion (STG); two other species (Cancer antenarius and Procambarus clarkii) do not. Further, the cholecystokinin-like immunoreactivity within the STGs of Panulirus and Homarus arise from distinct structures; from a projection of anterior ganglia in Panulirus, and from somata within the posterior motor nerves in Homarus. The staining in the other ganglia of the stomatogastric nervous system also shows some interspecies variability, although it appears to be more highly conserved than staining within the STG. These differences in staining were confirmed by measuring the amount of CCK-like peptide present in tissue extracts of ganglia by radioimmunoassay. In contrast to the variable staining within the STG, all four species have cholecystokinin-like immunoreactivity within the neurosecretory pericardial organs and thoracic segmental nerves. This cholecystokinin-like immunoreactivity is contained within fibers and within varicosities that coat the surface of these structures. The location of this staining and the presence of detectible levels of CCK-like peptide in the haemolymph suggests that CCK-like peptides in decapod crustacea may be utilized as neurohormones.
A simple relationship between the burst length of the trigger neurons of the Crustacea cardiac ganglion and the length of the heartbeat, is shown to lead to chaotic heart rate. Interestingly, the same type of relationship is also capable of generating complex periodic deviations from steady heart rhythm. Real Crustacea hearts are not likely to follow strictly either suite. The general procedure of analysis, however, is applicable to various biological rhythms, and explains how variation around the mean value can occasionally show up as intricate patterns which repeat themselves with accuracy.
Analysis of data obtained from molecular hybridization of 3H-labeled repetitious DNA has been utilized to reconstruct the broad outlines of phylogenetic relationships among decapod Crustacea. This molecular reconstruction agrees reasonably well with the paleontological record, and with other schemes obtained by comparative morphological and serological approaches. Preliminary evidence is in line with the hypothesis that continuous addition of new repeated sequence families to the genome over long periods of time may in part account for the correlation observed between percent repetitious DNA hybridized and divergence time. It is tentatively concluded that a core of DNA base sequence homology has been highly conserved throughout the evolution of the Crustacea. Demonstration of inter-species sequence homology has important implications to models which relegate a genetic regulatory function to repeated DNAs.
The oogenesis of the Crustacea Amphipoda Orchestia gammarellus can be divided in five stages taking into consideration both the oocyte ultrastructure and the physiology of the ovary. The primary oogonium (12 micron in diameter) is lodged within the germinative zone: after division, the daughter cell (or secondary oogonium) leaves this area and enters meiotic prophase. Stage I is represented by the oocyte with visible chromosomes (12-18 micron in diameter) the cytoplasmic ultrastructure of which is comparable to that of the oogonium. Stage II or previtellogenesis is characterized by a considerable growth of the oocyte (18-80 micron in diameter) which becomes enriched in ribosomes and vesicles of the rough endoplasmic reticulum; the oocyte does not yet contain any vitelline reserve (proteinaceous and lipid). Stage III or primary vitellogenesis (80-160 micron in diameter) is typified by the synthetic activity of the rough endoplasmic reticulum, corresponding to an endogenous accumulation of proteinaceous yolk. Stage IV or secondary vitellogenesis (160-800 micron in diameter) only appears during the period of reproduction; by means of endocytosis the oocyte accumulates yolk spheres in addition to lipid droplets, the origin of which is uncertain; towards the end of vitellogenesis, cortical granules become a feature that is noted for the first time in Crustacea. The last stage or maturation (800 micron in diameter) starts right before or immediately after the exuviation of the female and ends with fertilization.
Concentrations of polycyclic aromatic hydrocarbons have been determined in sediments, mussels and crustacea in the vicinity of a former gasworks site by Shoreham Harbour, UK. Very high concentrations of PAH were found in the substrate, an ash-like material deposited on the former gasworks site, which exhibited a profile consistent with the major source of contamination being coal or coke tar produced during the period of gas production at the site. Elevated PAH concentrations were also found in mussels both from the beach below the former gasworks site, and from sites further to the east in Portslade and Hove. The significance of these concentrations were assessed using an approach which involved the calculation of benzo[a]pyrene equivalent conoentrations (BaPEs), summing concentrations of individual PAH on the basis of their comparative potency as carcinogens. BaPE ranged from values of, or close to, zero for crustacea, to 336 microg kg(-1) wet weight in mussels from Southwick Beach. The contaminated mussels are not exploited commercially but may be taken by casual gatherers, and notices have been posted to warn potential consumers.
Since the last review of this topic, further insight has been gained into the presence and functions of cytochrome P450 proteins in the hepatopancreas and other organs of aquatic crustacean species, although progress has been slow relative to the advances in other species. Recent studies with several lobster, shrimp, crab and crayfish species suggest that cytochromes P450 in the 2 and 3 families are the most abundant forms in hepatopancreas microsomes. Substrates normally metabolized by CYP2 and CYP3 family members are monooxygenated more rapidly by crustacea than substrates normally metabolized by CYP1 family enzymes, e.g. erythromycin, testosterone and aminopyrine are much more rapidly monooxygenated than ethoxyresorufin. Some progress has been made in cloning and sequencing crustacean P450 forms. CYP2L1 and CYP2L2 cDNA sequences have been cloned from spiny lobster hepatopancreas libraries, and there was evidence for at least two more cytochromes P450 in spiny lobster hepatopancreas. An area of continued interest, but of no consensus or general findings, relates to the presence and inducibility of CYP1 family members in crustacea. Some studies indicate weak induction of total cytochrome P450 and increased turnover of substrates normally associated with CYP1, while others show no effect of the classic inducers that act at the Ah receptor in vertebrates. A few studies of the roles of cytochromes P450 in the biosynthesis and degradation of steroids, including ecdysteroids, have been published. Further studies are needed to understand the regulation and normal function of the crustacean cytochromes P450.
In the populations of aquatic troglobitic Crustacea, breeding females are found all year round, with an annual peak. Data about their biological cycle suggest four phases of reproduction per year. Synchronisation of egg-laying (for the fraction of females actually reproducing) seems to happen for several species of Crustacea both in nature and at the laboratory. The onset of reproduction is sharpest either late winter or mid-spring, and its amplitude decreases along the annual cycle. The existence of a seasonal trigger is probable.
Inside the labrum of Leptestheria dahalacensis are situated three types of large epidermal gland cells, whose ducts open onto the outer dorsal surface of the labrum. SEM revealed that the thin ducts of the A-type gland cells open out behind the epipharynx at the end of small, conically shaped protuberances, the two paired ducts of the B-type gland cells lead into the distal portion of the labrum, and the external opening of the single duct of the C-type gland cells lies on the dorsal lobe of the labrum. The ducts of the three different gland cell types have the same fundamental constitution, but vary in diameter. Each secretory unit consists of a pair of gland cells (A, B, or C) and a secretory duct. The duct is formed by ring-shaped folding of one anteroposteriorly elongated epidermal cell (duct cell), whose ends adhere closely to one another. A further ring-folded epidermal cell (accessory cell), but flattened in shape, is interposed, like a sleeve-connection, between the gland cells and the duct cell. The reservoirs of gland cells open into the lumen of the duct. Discontinuous deposits of highly electron-dense matter are present on the plasma membrane of the accessory cell delimiting the initial part of the duct lumen, while the plasma membrane of the duct cell facing the lumen is cuticularized. The cytoplasm of the accessory cell, on examination by TEM, appears quite similar to that of the duct cell, except for the different distribution and greater abundance of microtubules. Similarly organized tricellular tegumental glands also commonly occur in other Crustacea, both Malacostraca and non-Malacostraca. Possible functions of secretions from the three different types of gland cells present in the labrum of L. dahalacensis are discussed.
The sequence and timing of morphological changes during envelope formation was followed in diapause eggs of Pontella mediterranea (Crustacea, Copepoda). The multilayer coat enveloping these eggs resulted from the exocytosis of 4 types of cortical vesicles that sequentially released their contents in the perivitelline space. These included small high-density vesicles (hDV) with electron-dense material, vesicles (V) with dense ring granules and a uniform matrix contained within the same compartment, large high-density (HDV) vesicles, and large moderately dense (MDV) vesicles. All of these cortical vesicles were present in newly spawned, fertilized eggs. Their exocytosis resulted from egg activation. One of these cortical vesicles (V) was similar in morphology to the intracisternal granules precursors of endogenous yolk. Intracisternal granules, characteristic of previtellogenic oocytes of many crustaceans, were present in previtellogenic oocytes of P. mediterranea but disappeared in later stages of oocyte development once yolk formation was completed. We discuss the role of cortical vesicles in the formation of the complex extracellular coat enveloping copepod diapause eggs.
The impact of tannins from the environmental vegetation naturally polluting Alpine mosquito breeding sites was experimentally investigated by studying the toxicity of tannic acid, a natural hydrolyzable tannin, on the nontarget crustacean fauna associated with culicine populations. Bioassays indicate that exposure to tannic acid at concentrations from 0.06 to 2.0 mM is more deleterious to Chydorus sphaericus, Diaptomus castor, and Eucypris fuscata, than to Daphnia pulex, Acanthocyclops robustus, and Eucypris virens. Histopathological investigations after treatment with tannic acid at concentrations from 0.125 to 0.500 mM reveal sequential degenerative patterns of the midgut epithelium depending on the taxon, duration of the treatment, and concentrations assayed. These differential toxic effects on Crustacea are compared with those previously observed in larval Diptera, in order to evaluate the plant tannins as potentially useful products in integrated mosquito management programs.
Testosterone metabolism by Neomysis integer (Crustacea; Mysidacea) was assessed to obtain initial data on its metabolic capacity. N. integer were exposed to both testosterone and [(14)C]testosterone. Identification of testosterone metabolites and endogenous steroids was performed using thin-layer chromatography and liquid chromatography with multiple mass spectrometry. Endogenous production of testosterone in mysids was detected for the first time. N. integer were exposed to testosterone and metabolized administered testosterone extensively. At least 11 polar testosterone metabolites (R(f,metabolite) < R(f,testosterone)), androstenedione, dihydrotestosterone, and testosterone were produced in vivo by N. integer. A sex-specific testosterone metabolism was also observed, although this observation requires further confirmation. The anabolic steroid beta-boldenone was also identified for the first time in invertebrates. The metabolic pathway leading to the formation of beta-boldenone remains unknown, since the steroidal precursor androstadienedione could not be detected. These results reveal interesting similarities in enzyme systems in invertebrate and vertebrate species. Alterations in steroid hormone metabolism may be used as a new biomarker for the effects of endocrine disruptors in invertebrates.