Anisakis simplex and Terranova sp.: Inhibition by larval excretory-secretory products of mitogen-induced rodent lymphoblast proliferation

Execretory-secretory products were collected from supernatants of in vitro cultures of larval nematodes, Anisakis simplex (type I) and Terranova sp. (Hawaii type A). These materials were found to be more potent inhibitors of rodent lymphocyte blast transformation induced by concanavalin A and bacterial lipopolysaccaride than whole worm extracts of the same parasites. Inhibition of blast transformation was a result of cytostatic rather than toxic effects on proliferating lymphoid cells. The material(s) responsible for suppression are greater than 10,000 MW and are heat labile.     

Identification of lipids in the cuticle of the parasitic nematode Anisakis simplex and the somatic tissues of the Atlantic cod Gadus morhua

The main aim of this work was to assign the cuticular lipids identified in a parasitic nematode and to distinguish those originating from its host. The hypothesis that long-chained fatty acids and sterols are imported by the parasite in the absence of certain enzymes was also tested. The organisms (Anisakis simplex and Gadus morhua) were extracted in petroleum ether and dichloromethane. Matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF) was used to identify unknown components, and electrospray ionization mass spectrometry (ESI/MS) to verify recognized groups of lipids. The lipid classes identified in the surface layer were free saturated and unsaturated fatty acids, triacylglycerols, sterols and non-polar sphingolipids (ceramides, sphingoid bases). The most abundant fraction consisted of fatty acids. The predominant saturated acids were tetradecanoic acid in the petroleum ether extract of A. simplex, hexadecanoic acid in the dichloromethane extract of A. simplex, and also the polyunsaturated octadecahexaenoic and octadecatrienoic acids in both extracts of the parasitic nematode. The mass spectrum revealed the presence of fatty acids with different numbers of carbons, and with odd and even numbers of unsaturated bonds. The MALDI-TOF mass spectrum also identified triacylglycerols (TAGs). The dominant short-chain TAGs were CoCoCy:₁, CoCoPg and Bu0:0B:₆. The majority of TAGs were found in the ether and dichloromethane extracts of A. simplex. Sterols were the least common class of lipids found in the nematode extracts; most likely, this is the fraction that is entirely incorporated from the host organism because of the parasite’s inability to synthesize them. MALDI-TOF also identified non-polar sphingolipids - ceramides and sphingoid bases. The signals due to N-octanoyl-d-erythro-octasphinganine (m/z 288.3) and N-tetranoyl-d-erythro-tetradecasphinganine (m/z 316.4) were dominant on the mass spectra; quite a large number of short-chain non-polar sphingolipids were also identified.     

鱼类Fc受体研究

Fc受体是免疫细胞表面一种重要受体分子,通过与免疫球蛋白Fc段结合触发多种生物学功能,是联系体液免疫和细胞免疫的桥梁。部分硬骨鱼中已经发现了Fc受体,在斑马鱼、斑点又尾鲴和鲤鱼中都克隆到了Fc受体的γ亚基,在鲨鱼和大西洋鲑中证明有能够与免疫球蛋白结合的Fc受体存在,并在斑点叉尾鲴、河豚和虹鳟中存在着类似α亚基的Fc受体。对鱼类Fc受体的发现和研究必将为了解鱼类的免疫机制及免疫进化提供重要的资料。     

The status of marine fish larval-rearing technology in Australia

Over 20 marine fish species have been studied forfarming or stock enhancement in Australia. However,commercial production has been dominated by the cageculture of Salmo salar in Tasmania, Thunnusmaccoyiiin South Australia, and to a lesser extent Latescalcarifer in Queensland. A major impediment to thecommercial production of new species has been thelarge-scale production of juvenile fish. Thedevelopment of marine fish larval rearing technologyin Australia has had four main influences over thelast decade: culture system technology from France,live food culture and nutritional enhancement fromBelgium, artificial diets from Japan and extensivepond culture from the USA. Microalgae and live foodculture is based on traditional aquaculture speciesand methods. Recent Australian research has focusedon induced spawning, the role of stress in inhibitingovulation, factors influencing initial swim bladderinflation in larvae, larval nutrition, extensiveculture and diagnosis of disease. Over the next 5years, Australian aquaculturists should be able toproduce industrial quantities of a range of nativemarine fish, either in intensive fish hatcheries, orin combination with extensive pond culture.     

Sterol composition of the marine sponge Crambe crambe

The sterol content of the marine sponge Crambe crambe has been determined. The major components of the mixture are cholest-7-en3β-ol, 24-methylcholesta-7,22-dien-3β-ol and cholesta-7,22-dien-3β-ol. Significative quantities of the rare 4α-methyl-5α-cholest-8-en3β-ol are also present.     

Sesquiterpenes from the marine red alga Laurencia distichophylla

A new cuparene-type sesquiterpene, isolaurenisol, has been isolated and identified from the New Zealand red alga Laurencia distichophylla. Major differences in the chemical composition of two morphologically indistinguishable samples of L. distichophylla are noted.     

Detrimental effects of Ulva lactuca (L.) exudates and low oxygen on estuarine crab larvae

Conventional theory postulates that associations between marine macrophytes and animals are generally positive. This paper presents evidence, however, that a common species of green macroalga, Ulva lactuca (L.), is detrimental to estuarine invertebrates due to the production of toxic exudates and low oxygen tensions which occur in the seaweed beds at night. Bioassays of the responses of zoeae of five species of estuarine crabs (Callinectes sapidus Rathbun, Carcinus maenas L., Eurypanopeus depressus Smith, Neopanope texana savi Smith and Rhithropanopeus harissii Gould), using water in which Ulva lactuca was cultured for 24 h, produced 100% mortality after 22 days. No crabs survived the molt into megalopa. Hypoxic water, 0.5 ± 0.3 ppm oxygen, caused a decline in larval activity (movement), but there was no mortality over an 8-h period, Ulva-water purged to 0.4 ± 0.1 ppm oxygen caused 100% mortality in 13–40 min. These synergistic effects could be critical in estuaries where dense U. lactuca beds cause periods of low dissolved oxygen. We hypothesize larval recruitment may be limited in such systems, particularly in areas where flushing is poor.     

Global patterns in marine dispersal estimates: the influence of geography, taxonomic category and life history

We examine estimates of dispersal in a broad range of marine species through an analysis of published values, and evaluate how well these values represent global patterns through a comparison with correlates of dispersal. Our analysis indicates a historical focus in dispersal studies on low-dispersal/low-latitude species, and we hypothesize that these studies are not generally applicable and representative of global patterns. Large-scale patterns in dispersal were examined using a database of correlates of dispersal such as planktonic larval duration (PLD, 318 species) and genetic differentiation (FST, 246 species). We observed significant differences in FST (p<0.001) and PLD (p<0.001) between taxonomic groups (e.g. fishes, cnidarians, etc.). Within marine fishes (more than 50% of datasets), the prevalence of demersal eggs was negatively associated with PLD (R2=0.80, p<0.001) and positively associated with genetic structure (R2=0.74, p<0.001). Furthermore, dispersal within marine fishes (i.e. PLD and FST) increased with latitude, adult body size and water depth. Of these variables, multiple regression identified latitude and body size as persistent predictors across taxonomic levels. These global patterns of dispersal represent a first step towards understanding and predicting species-level and regional differences in dispersal, and will be improved as more comprehensive data become available.     

Experimental studies on the migration of Anisakis sp. larvae (Nematoda: ascaridida) into the flesh of herring, Clupea harengus L.

In two experiments, a batch of 38 herring from Shetland was gutted immediately on capture (0 h) and further batches were gutted 14 and 37 h after capture and storage on ice. The numbers of Anisakis larvae in the body-cavity and in the flesh (detected by pepsin-HCl digest) were then counted. The proportion of fish with worms in the flesh was higher at 14 and 37 h than at 0 h. The percentage of the total worm burden in the flesh increased between cash each time interval, which suggests that a large-scale migration of larvae into the flesh occurs in ungutted fish. There was a positive association between the numbers of larvae in the viscera and the numbers in the flesh. Most larvae occurred in the hypaxial muscles. The results are discussed in relation to the observations of earlier workers.     

The occurrence of the second moult of Ascaris lumbricoides and Ascaris suum

Maung M. 1978. The occurrence of the second moult of Ascaris lumbricoides and Ascaris suum. International Journal for Parasitology 8: 371–378. Eggs of Ascaris lumbricoides and A. suum were cultured at 28°C and observed daily. Larvae were released by pressure, by artificial hatching with CO₂, and by natural hatching after infection of laboratory mice. The early stages of development in the egg were observed to comprise two moults, one occurring immediately after the other. Both moults were initiated within the egg, but the time of completion of the second moult varied considerably, and in some instances was not completed until the larvae reached the liver of experimentally infected animals.     

The brain can eat: Establishing the existence of a central pattern generator for feeding in third instar larvae of Drosophila virilis and Drosophila melanogaster

To establish the existence of a central pattern generator for feeding in the larval central nervous system of two Drosophila species, the gross anatomy of feeding related muscles and their innervation is described, the motor units of the muscles identified and rhythmic motor output recorded from the isolated CNS. The cibarial dilator muscles that mediate food ingestion are innervated by the frontal nerve. Their motor pathway projects from the brain through the antennal nerves, the frontal connectives and the frontal nerve junction. The mouth hook elevator and depressor system is innervated by side branches of the maxillary nerve. The motor units of the two muscle groups differ in amplitude: the elevator is always activated by a small unit, the depressor by a large one. The dorsal protractors span the cephalopharyngeal skeleton and the body wall hence mediating an extension of the CPS. These muscles are innervated by the prothoracic accessory nerve. Rhythmic motor output produced by the isolated central nervous system can simultaneously be recorded from all three nerves. The temporal pattern of the identified motor units resembles the sequence of muscle contractions deduced from natural feeding behavior and is therefore considered as fictive feeding. Phase diagrams show an almost identical fictive feeding pattern is in both species.     

The escape response of food-deprived cod larvae (Gadus morhua L.)

The escape response of Atlantic cod larvae (Gadus morhua) 25 and 47 days post hatch (dph) - either fed or deprived of food for three days - was studied. Larval escape responses were provoked by water movement from the suction of a fixed-position pipette. Escape latency, distance, speed, burst speed, and vertical and lateral escape angles were quantified using motion tracking software designed for 3-D silhouette video recordings. Escape performance, expressed as escape distance and escape speed, improved with age. The escape angles were normally distributed and highly variable, ranging from − 170° to 170° and − 40° to 105° for lateral and vertical escape angles respectively. No food deprivation-induced effects in any of the behaviours were found, suggesting that there are no condition-related behavioural effects (size-independent effects) in escape response performance after 3 d of food deprivation. This may reflect a negligible difference in the cost/benefit equation for fed vs. food-deprived larvae in performing an escape response when under attack.     

Intact antigen uptake in intestinal epithelial cells of marine fish larvae

Endocytosis of bacteria by epithelial cells in the hindgut of 4- to 6-day-old cod ( Gadus morhua L. ) larvae and of 10- to 12-day-old herring ( Clupea harengus L. ) larvae was demonstrated by fluorescence microscopy and transmission electron microscopy. Fixed sheep erythrocytes and bacteria-sized latex particles were not ingested by these cells. By immunohistochemical methods intact bacterial antigens were observed in columnar epithelial cells in the foregut of 4-day-old yolk-sac larvae of cod. In these cells a preferential uptake of Vibrio fischeri and Vibrio salmonicida cells as compared to Flavobacterium sp. was found. The possible role of these findings in the establishment of an indigenous microflora, defence against pathogens or nutrient absorption in fish larvae is discussed.     

Cold tolerance in diapausing and non-diapausing stages of the flesh fly, Sarcophaga crassipalpis

ABSTRACT. Supercooling points (SCP) and low temperature tolerance were determined for larval, pupal and adult stages of Sarcophaga crassipalpis Macquart (Diptera: Sarcophagidae). No stage tolerates tissue-freezing. Ontogenetic changes in SCP profiles are similar for comparable developmental stages of diapause and non-diapause groups. Feeding larvae have SCPs near -7°C which decrease to -11°C in the postfeeding wandering phase of the final larval instar. The lowest SCPs are recorded for pupae at -23°C. The capacity to survive at -17°C varies with age of the diapausing pupae: 10-day-old pupae are less cold tolerant than pupae that have been in diapause for 45–80 days. Although the SCP of non-diapausing pupae is as low as in diapausing pupae, non-diapausing pupae are extremely sensitive to low temperature exposure and do not survive to adult eclosion when exposed to -17°C for as little as 20 min. The use of hexane to break pupal diapause has no effect on SCPs or low temperature tolerance.     

The attraction of larvae of Culex pipiens quinquefasciatus Say to ribonucleic acid and nucleotides

The positive chemotaxis of larvae of Culex pipiens quinquefasciatus in response to RNA or its degradation products has been demonstrated. It has further been shown that larvae are specifically attracted to certain nucleoside monophosphates. The observations are related to the known dietary requirements of mosquito larvae.     

Encapsulation response of the marine prosobranch Cerithidea californica to natural infections of Renicola buchanani sporocysts (Trematoda: Renicolidae)

A hyalinocyte-mediated encapsulation reaction is elicited by sporocysts of Renicola buchanani infecting the anterior mantle region of Cerithidea californica. Three phases of capsule formation are recognized: (1) an initial aggregating of hyalinocytes around each sporocyst in which pseudopodia from encapsulating cells loosely interdigitate with the parasite's tegumental microvilli, (2) the infiltrating of numerous hyalinocytes to form a dense matrix of cells which lies in close contact with the sporocyst's tegument, and (3) the horizontal flattening of hyalinocytes against the sporocyst's surface to form a tightly adhering capsule four to eight cell layers deep. Sporocysts are not harmed as a consequence of encapsulation. Capsule formation in response to R. buchanani sporocysts is considered a type of leucocytic encapsulation, specifically designated hyalinocytic encapsulation.     

See the light: Electrophysiological characterization of the Bolwig organ's light response of Calliphora vicina 3rd instar larvae

The anatomy and development of the larval cyclorraphous Diptera visual system is well established. It consists of the internal Bolwig organ (BO), and the associated nerve connecting it to the brain. The BO contributes to various larval behaviors but was never electrophysiologically characterized. We recorded extracellulary from the Bolwig nerve of 3rd instar Calliphora vicina larvae to quantify the sensory response caused by BO stimulation with light stimuli of different wavelengths, intensities and directions. Consistent with previous behavioral experiments we found the BO most sensitive to white and green, followed by blue, yellow, violet and red light. The BO showed a phasic-tonic response curve. Increasing light intensity produced a sigmoid response curve with an approximate threshold of 0.0105 nW/cm² and a dynamic range from 0.105 nW/cm² to 52.5 nW/cm². No differences exist between feeding and wandering larvae which display opposed phototaxis. This excludes reduced BO sensitivity from causing the switch in behavior. Correlating to the morphology of the BO frontal light evoked the maximal reaction, while lateral light reduced the neural response asymmetrically: Light applied ipsilaterally to the recorded BO always produced a stronger response than when applied from the contralateral side. This implies that phototacic behavior is based on a tropotactic mechanism.     

Viral abundance and genome size distribution in the sediment and water column of marine and freshwater ecosystems

The size distribution of viral DNA in natural samples was investigated in a number of marine, brackish and freshwater environments by means of pulsed field gel electrophoresis (PFGE). The method was modified to work with both water and sediment samples, with an estimated detection limit for individual virus genome size groups of 1-2 x 10(4) virus-like particles (VLP) mL(-1) water and 2-4 x 10(5) VLP cm(-3) sediment in the original samples. Variations in the composition and distribution of dominant virus genome sizes were analyzed within and between different habitats that covered a range in viral density from 0.4 x 10(7) VLP mL(-1) (sea water) to 300 x 10(7) VLP cm(-3) (lake sediment). The PFGE community fingerprints showed a number of cross-system similarities in the genome size distribution with a general dominance of genomes in the 30-48, 50-70 and 145-200 kb size fractions, and with many of the specific genome sizes detected in all the investigated habitats. However, large differences in community fingerprints were also observed between the investigated sites, and some virus genome sizes were found only in specific biotopes (e.g. lake water), in specific ecosystems (e.g. a particular lake) or even in specific microhabitats (e.g. a particular sediment stratum).