A Glycosylated Byssal Precursor Protein from the Green Mussel Perna viridis with Modified Dopa Side-chains

Foot tissue of the green mussel Perna viridis contains a variety of byssal precursor proteins with the unusual redox-active amino acid, Dopa (L-β-3,4-dihydroxyphenyl-α-alanine). Eight proteins were detectable in acidic extracts of the Perna foot by a redox cycling assay with nitroblue tetrazolium. In one of these, however, P. viridis foot protein-1 (Pvfp-1), activity was not due to Dopa, but to another redox-active derivative. Based on specific colorimetric derivatization with Arnow's reagent, ninhydrin and phenylisothiocyanate (Edman), mass spectrometry, the redox-active derivative in Pvfp-1 is not consistent with any known modification. Another uncommon modification of Pvfp-1 involves O-glycosylation of threonine by mannose, glucose or fucose. As in previously characterized fp-1s, the primary sequence of the Pvfp-1 (apparent mass 89?kDa) has two consensus decapeptide motifs; one is APPKPX₁TAX₂K and the other is APPPAX₁TAX₂K, where P is Pro/Hyp, and X₁ and X₂ are difucosylated threonine and a redox sensitive derivative of tyrosine or Dopa, respectively. Of these two unusual residues, X₂ is unique to Pvfp-1, whereas O-glycosylated Thr has been previously detected in freshwater mussel fp-1. The sequence homology of Pvfp-1 with the common structural motifs of the fp-1 protein family strongly suggests that the Pvfp-1 functions as the byssal coating (lacquer) protein.     

Feeding rhythms of the green-lipped mussel, Perna viridis (Linnaeus, 1758) (Bivalvia: Mytilidae) during spring and neap tidal cycles

Food availability and feeding responses of the green mussel Perna viridis were investigated for two complete tidal cycles during both spring and neap tides. Temporal changes in total particulate matter (TPM), particulate inorganic matter (PIM) and particulate organic matter (POM), were smaller during neap than spring tides. Seston characteristics at different times of a tidal cycle were compared for both spring and neap tides. Only during spring tides were TPM and PIM significantly higher at high tides while POM remained relatively constant (P>0.05). The clearance rate of the mussels underwent temporal variations with tides, and was a negative power function of TPM and a positive linear function of f (organic content), during both spring and neap tides. f was the key factor influencing filtration rate, organic ingestion rate, absorption rate and absorption efficiency. All feeding rates increased linearly with increases in organic content. Pseudofaeces were produced only during spring but not neap tides. Feeding rates and absorption efficiency were highest at low and lowest at high tides (P<0.01). There was no significant temporal change in the wet weight and protein content of the crystalline style with the tidal regime. For the digestive gland, alpha-amylase activity was higher at spring than at neap tides, and higher during high tides in a tidal cycle. The digestive gland cellulase activity did not change significantly with the tides. For the crystalline style, both the activity of cellulase and alpha-amylase were not significantly different (P>0.05) between spring and neap tides. Tidal rhythms in feeding and digestion in this species were likely controlled by temporal variations in food availability in the seawater. By adjusting feeding rates and enzymatic activities, absorption in Perna viridis remained constant, irrespective of the changes in food availability.     

Spatial and Temporal Genetic Variation of Green Mussel, Perna viridis in the Gulf of Thailand and Implication for Aquaculture

The culture of green mussel (Perna viridis) in the Gulf of Thailand depends on natural spat which are believed to come from spawning grounds adjacent to major river mouths. In the present paper, genetic diversity of spatial and temporal populations of green mussel in the Gulf of Thailand was investigated using five microsatellite loci. The results showed moderate genetic variation of all 11 populations (averaged number of alleles per locus, A = 10.4–12.2; effective number of alleles per locus, A _e = 5.36–6.59; mean allelic richness, A _r = 10.23–12.06; observed heterozygosity, H _o = 0.52–0.63, and expected heterozygosity, H _e = 0.66–0.73) without significant differences among populations. No sign of bottleneck or genetic disequilibrium was observed. Genetic differentiation among spatial populations was low (F _ST = 0.0046, CI_0.95 = 0.0020–0.0083 for the samples collected in January, 2007, and F _ST = 0.0088, CI_0.95 = 0.0010–0.0162 for the samples collected in July, 2007) while temporal variation was significant as revealed by the analysis of molecular variance. Multidimensional scaling separated temporal population groups with minor exception. The assignment test revealed that most of the recruits were from other populations.     

In Vitro Antibacterial and Antioxidant Studies of Croton roxburghii L., from Similipal Biosphere Reserve

In vitro antibacterial activities of acetone, ethanol, methanol and water extracts of leaves and bark of Croton roxburghii L. studied against ten human pathogenic bacterial strains showed significantly higher activity in acetone extract and least activity in case of aqueous. Minimal inhibitory concentration (MIC) values of all extracts ranged between 0.62 and 10 mg/ml, while minimal bactericidal concentration (MBC) values ranged from 1.25 to values greater than 10 mg/ml. The antioxidant assays viz. DPPH, hydrogen peroxide scavenging, iron reducing and iron chelating assays along with total phenol and ascorbic acid content were carried out with aqueous extracts of leaves and bark. While the total phenol contents in leaves and bark extracts were 0.766 ± 0.014 and 0.735 ± 0.028% respectively their ascorbic acid contents were found to be 0.252 ± 0.019 and 0.431 ± 0.013% respectively. DPPH activities in both (leaves and bark) extracts increased with the increase in concentrations. Iron chelating capacity of leaves extract is significantly higher than that of the bark. Leaves extract showed an increase in percentage of scavenging property with the increase in concentrations. Plant extracts showed low amount of iron reducing property at all concentrations. Hydrogen peroxide scavenging properties of bark was low than that of the leaves.     

Effects of Total Soft Tissue and Shell Thickness on the Accumulation of Heavy Metals (Cd,Cu, Pb,and Zn) in the Green-Lipped Mussel Perna viridis

The effects of the total soft tissue dry weight and shell thickness and on the accumulation of Cd, Cu, Pb, and Zn were determined in the green-lipped mussel Perna viridis. In agreement with Boyden's formula (1977), our results showed that the plotting of metal concentrations against the total soft tissue dry weight and shell thickness of the mussel on a double logarithmic basis gave negative coefficients especially for Cd, Pb, and Zn. Therefore, the smaller mussels (lower total soft tissue dry weight) had higher concentrations of Cd, Pb, and Zn than the larger ones. Since shell thickness could be considered to estimate of the age of the mussel, it was also found that the younger mussels accumulated more Cd, Pb, and Zn than the older ones. This indicated that P. viridis has a different metabolic strategy for each of the metals studied which may be related to age. However, the accumulation of Cu was hardly affected by the sizes and ages of the mussel. This indicated that the accumulation pathways of Cu and the processes affecting the bioavailability of Cu to the mussel are different from those for Cd, Pb, and Zn.     

Non-Lethal Heat Shock of the Asian Green Mussel,Perna viridis,Promotes Hsp70 Synthesis,Induces Thermotolerance and Protects Against Vibrio Infection

Mild heat stress promotes thermotolerance and protection against several different stresses in aquatic animals, consequences correlated with the accumulation of heat shock protein 70 (Hsp70). The purpose of this study was to determine if non-lethal heat shock (NLHS) of the Asian green mussel, Perna viridis, an aquatic species of commercial value, promoted the production of Hsp70 and enhanced its resistance to stresses. Initially, the LT₅₀ and LHT for P. viridis were determined to be 42°C and 44°C, respectively, with no heat shock induced death of mussels at 40°C or less. Immunoprobing of western blots revealed augmentation of constitutive (PvHsp70-1) and inducible (PvHsp70-2) Hsp70 in tissue from adductor muscle, foot, gill and mantel of P. viridis exposed to 38°C for 30 min followed by 6 h recovery, NLHS conditions for this organism. Characterization by liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed that PvHsp70-1 and PvHsp70-2 respectively corresponded most closely to Hsp70 from P. viridis and Mytilus galloprovincialis. Priming of adult mussels with NLHS promoted thermotolerance and increased resistance to V. alginolyticus. The induction of Hsp70 in parallel with enhanced thermotolerance and improved protection against V. alginolyticus, suggests Hsp70 functions in P. viridis as a molecular chaperone and as a stimulator of the immune system.     

Accumulation, depuration and distribution of cadmium and zinc in the green-lipped mussel Perna viridis (Linnaeus) under laboratory conditions

Ecotoxicological tests were conducted in the green-lipped mussel Perna viridis under laboratory conditions. Different rates of accumulation and depuration in soft tissues are found and this might be due to different mechanisms of metal binding and regulation. At the end of depuration, Cd levels in soft tissues of P. viridis were 10–30 times higher than before exposure, while Zn levels in soft tissues were almost similar to levels before exposure. These results indicate that P. viridis is a good biomonitoring organism for Cd but Zn levels might be actively regulated. It remains uncertain whether P. viridis is a good biomonitoring organism of environmental Zn contamination. However, the positive patterns, although different rates, of accumulation and depuration for Cd and Zn support the use of P. viridis as a biomonitoring agent for such metals.     

Effect of zinc and benzene on respiration and excretion of mussel larvae (Perna perna) (Linnaeus, 1758) (Mollusca; Bivalvia).

The presence of pollutants in the ocean may affect different physiological parameters of animals. Oxygen consumption and ammonia excretion were evaluated in D-shaped larvae of mussels (Perna perna) exposed to zinc sulphate (ZnSO(4)) and benzene (C(6)H(6)). When compared to the control group, both pollutants presented a significant reduction in oxygen consumption. A reduction in the ammonia excretion was also observed, both for ZnSO(4) and C(6)H(6) and also in the oxygen consumption. The results indicate that anaerobic metabolism may occur at the beginning of P. perna mussels development, as observed in veliger larvae. The O:N ratio under experimental conditions showed low values indicating that catabolism in veliger larvae was predominantly proteic.     

If the Asian green mussel,Perna viridis (Linnaeus, 1758), poses the greatest invasive marine species threat to Australia,why has it not invaded?

A national approach has been developed to the problem of invasive marine species (IMS) in the Australian marine environment. Fifty-five species were listed as posing significant threats to Australia. A 2005 analysis of the scientific literature concluded that the Asian green mussel Perna viridis (Linnaeus, 1758) poses the greatest threat to Australia. The mussel has in fact successfully invaded many areas of the world’s oceans. Despite the numerous and varied opportunities for P. viridis to be distributed to northern Australia it has not established a known population on the continent, perhaps suggesting there are biological factors inhibiting its establishment. The invasion success of P. viridis in many parts of the world and its failure so far to establish in Australia make the species ideal for testing theories of the factors determining invasion success. Such research will allow a reconsideration of the invasion threat the species poses to the Australian marine environment.     

The spawning, embryonic and early larval development of the green wrasse Labrus viridis (Linnaeus, 1758) (Labridae) in controlled conditions

Green wrasse, Labrus viridis (Linnaeus, 1758), is an endangered species in the southern Adriatic Sea, but it is also of interest for potential rearing in polyculture with other commercial species for the repopulation of areas where it is endangered or as a new aquaculture species. A parental stock of the green wrasse was kept in aquaria for six years. The spawning, embryonic and early larval development maintained under controlled laboratory conditions are described and illustrated. The average diameter of newly spawned eggs was 1.01±0.03 mm. Mature and fertilized eggs were attached to the tank bottom by mucus. Hatching started after 127 h at a mean temperature of 14.4±0.8°C. The average total length of newly hatched larvae was 4.80±0.22 mm. Absorption of the yolk-sac was completed after the 5th day when larvae reached 5.87±0.28 mm. Larvae were fed with the rotifers Brachionus plicatilis. The pigmentation of L. viridis larvae is similar to that of Labrus merula and Labrus bergylta, but the main differences between these species are in the size of larvae and the development time of the melanophores on the anal fin-fold (five days later than with L. merula) and on top of the head (nine days earlier than with L. merula).     

Studies of Inositol Phosphate Export from Neuronal Tissue In Vitro

Abstract: Recent in vivo microdialysis studies have demonstrated the presence of extracellular levels of inositol 1,4,5-trisphosphate [Ins(1,4,5)P₃] that can be increased in a concentration-dependent manner by muscarinic receptor activation. The aim of the present study was to determine whether extracellular levels of Ins(1,4,5)P₃ could be measured in vitro. Despite rapid increases in internal Ins(1,4,5)P₃ levels after stimulation with 1 m M carbachol, there was no change in external levels in both rat brain cortical slices and human neuroblastoma SH-SY5Y cells. Suprafusion of myo -[³H]inositol-prelabelled hippocampal slices with 1 m M carbachol caused an increase in ³H-inositol phosphates over basal levels in the perfusate after 10 min, reaching a peak (223 ± 56% of basal) 20 min after suprafusion with carbachol was started. This response to carbachol was potentiated in the presence of 30 m M K⁺. Analysis of the individual ³H-inositol phosphates in the perfusate revealed that levels of [³H]inositol monophosphate, [³H]inositol bisphosphate, [³H]inositol trisphosphate, and [³H]inositol tetrakisphosphate were all significantly increased. A similar increase in extracellular ³H-inositol phosphates was demonstrated in SH-SY5Y cells incubated with 1 m M carbachol for 30 min. This response was again enhanced by 30 m M K⁺, although the intracellular response was not potentiated. Possible roles for extracellular inositol phosphates are discussed.     

DNA isolation of mucus from Salmo trutta (Linnaeus, 1758) and Thymallus thymallus (Linnaeus, 1758) as an alternative method to conventional fin‐clipping

The first objective of the actual project was to investigate if mucus from Salmo trutta (Linnaeus, 1758) and Thymallus thymallus (Linnaeus, 1758) is suitable for DNA extraction. In the second step, it was analyzed if the avoidance of environmentally hazardous chemicals could lead to sufficiently pure DNA isolation. Finally, it was examined if samples taken from different fish body parts yielded different grades of purity of the extracted DNA. Mucus was collected from 552 individuals of S. truttaand T. thymallusfrom 2012 to 2015. DNA was extracted with two different kits (E.Z.N.A.^® Insect DNA Kit and my‐Budget DNA Mini Kit) and two different swabs (my‐Budget roughened laminated sterile cotton swabs and common autoclaved cotton swabs). The results showed that the my‐Budget DNA Mini Kit in combination with the my‐Budget roughened laminated sterile cotton swabs was the most suitable method. It is a non‐destructive and swift technique that results in the concentration of DNA being high and pure.     

Novel microsatellite DNA loci for Bombus terrestris (Linnaeus, 1758)

We present details and characteristics of 123 novel polymorphic microsatellite DNA loci for Bombus terrestris. Thirty-four of these loci have been tested in nine other Bombus species and 25 of them showed polymorphisms in at least one species. These microsatellite DNA loci together with the already established 60 loci will be useful for characterizing wild and managed populations of B. terrestris and other Bombus species as well as for detailed genetic studies in including mapping studies and genome annotations.     

Isolation and characterization of polymorphic microsatellites from Asian green mussel (Perna viridis)

Asian green mussels (Perna viridis) belonging to the family Mytilidae are native to the coastal and tropical marine waters of the Indo‐Pacific region. Ten polymorphic microsatellites were isolated and their polymorphisms were determined in 36 individuals. The average allele number of the 10 microsatellites was 11.7/locus with a range of two to 24 and the expected heterozygosity averaged at 0.69, ranging from 0.18 to 0.94. Eight out of 10 microsatellites agreed with the Hardy–Weinberg equilibrium and showed independent segregation. These microsatellites will facilitate the studying of the genetic diversity and population structure of the green mussel in and outside of the Indo‐Pacific region of Asia.     

Testing baits prepared from by‐product of the shrimp and snow crab industry in the pot fishery for Gadus morhua (Linnaeus, 1758) and Pollachius virens (Linnaeus, 1758)

In a study between October and November 2015 in northern Norway, bait prepared from the by‐product of the shrimp and snow crab industry was employed in the pot fishery to capture cod and saithe. Frozen natural herring was used as control bait. Pots with the three bait types were placed 1,000 m apart in order to eliminate the risk of interaction between the different baits. In total 11 sampling periods were conducted. There were no significant differences in catch rates between the different baits tested on cod and saithe. The manufactured bait tested in this study showed a continuous release of odours over a 14‐day period. Results show that the water content of the bait increases rapidly in the first phase of the immersion (until day 7), and the corresponding loss of bait mass (dry weight) suggest some physical disintegration (erosion) of the bait until the water content stabilizes. This study illustrates that it is possible to replace natural herring with hydrolysates from by‐products from shrimp or snow crab industry in the Norwegian pot fishery of cod and saithe.     

The Medusa of Velella velella (Linnaeus, 1758) (Hydrozoa, Chondrophorae)

A mature medusa of Velella velella (Linnaeus, 1758) is reportedfor the first time from the North Atlantic; previously adultmedusae were known only from the Mediterranean. This specimen,collected by SCUBA divers, is the largest specimen recordedto date. Distinctive features are: two opposite adaxial-axialpairs of perradial capitate tentacles; two marginal bulbs lackingtentacles; conical manubrium with 4 perradial longitudinal gonads;perradial exumbrellar cnidae tracts; cnidome of stenoteles andmacrobasic euryteles; and zooxanthellae within the subumbrella.Since this specimen was collected near the surface and has zooxanthellae,it is likely that V. velella medusae are epipelagic.     

Influence of salinity on the physiological conditions in mussels, Perna perna and Perna viridis (Bivalvia: Mytilidae)

Perna genus was introduced to Venezuela, but nowadays, Perna perna and Perna viridis coexist and are commercially exploited from their natural beds. The aim of this work was to determine the effect of salinity on the physiological conditions of these species by studying RNA/DNA and Protein/DNA ratios. The organisms were collected from natural beds at La Esmeralda, Sucre State, Venezuela, and acclimatized for 15 days under laboratory conditions at 25 degrees C, 36 per thousand salinity, pH between 7 and 8 and more than 90% of oxygen saturation. Later, they were divided in two groups: for one group, the salinity concentration was increased (36 to 45 per thousand), and for the other, the salinity was decreased (36 to 15 per thousand). The rate of change was 1 per thousand every day. Ten organisms per group of both species were taken at each of 15, 20, 25, 30, 36, 40 y 45 per thousand salinity concentrations. Protein (colorimetric method) and nucleic acids (RNA and DNA by fluorometric method) concentrations were measured in the digestive gland, gills and adductor muscle tissues. Results indicate that P. perna can physiologically compensate the increase in salinity, but not when the salinity decreased, when proteins are the most needed macromolecules. The Protein/DNA index is directly related to salinity changes in both cases. P. viridis shows physiological compensation to salinity increases and decreases. The RNA/DNA index value in both cases supports this. Digestive gland and muscle tissues are the regulating tissues in both species. These results show that P. viridis has a higher degree of adaptability to salinity changes and, therefore, a greater potential for aquaculture than P. perna.     

中国翼手目新记录——马来假吸血蝠

2006年11月26日在云南省勐腊县勐仑镇翠鼻峰一无名洞捕捉到1只雄性蝙蝠,体型中等,前臂长60.7 mm,体重21.4 g。耳大,椭圆形,两耳于前额上方约15%的高度相连,耳屏细长双叉;鼻叶较简单,后鼻叶椭圆形,顶部钝圆、两侧缘向外隆凸,中央具一条显著的纵形隆脊,隆脊下部与隆起的间鼻叶相接,间鼻叶呈三角形,顶部W-形,前鼻叶呈马蹄状,前端紧接吻部;无尾,第二指仅具第一指骨;上门齿缺失,上犬齿前后基部均有一较大的副尖,Pm³缺失,下颚第二前臼齿（Pm₃）缺失。经鉴定,该蝙蝠为中国新记录——马来假吸血蝠（Megaderma spasma）。该文给出此蝙蝠的形态与头骨测量数据,并与国内分布的另一种假吸血蝠——印度假吸血蝠（M. lyra）进行了对比。标本保存在广东省昆虫研究所。