Background: Some of the most widely recognized coral reef fishes are clownfish or anemonefish, members of the family Pomacentridae (subfamily: Amphiprioninae). They are popular aquarium species due to their bright colours, adaptability to captivity, and fascinating behavior. Their breeding biology (sequential hermaphrodites) and symbiotic mutualism with sea anemones have attracted much scientific interest. Moreover, there are some curious geographic-based phenotypes that warrant investigation. Leveraging on the advancement in Nanopore long read technology, we report the first hybrid assembly of the clown anemonefish (Amphiprion ocellaris) genome utilizing Illumina and Nanopore reads, further demonstrating the substantial impact of modest long read sequencing data sets on improving genome assembly statistics.
Results: We generated 43 Gb of short Illumina reads and 9 Gb of long Nanopore reads, representing approximate genome coverage of 54× and 11×, respectively, based on the range of estimated k-mer-predicted genome sizes of between 791 and 967 Mbp. The final assembled genome is contained in 6404 scaffolds with an accumulated length of 880 Mb (96.3% BUSCO-calculated genome completeness). Compared with the Illumina-only assembly, the hybrid approach generated 94% fewer scaffolds with an 18-fold increase in N50 length (401 kb) and increased the genome completeness by an additional 16%. A total of 27 240 high-quality protein-coding genes were predicted from the clown anemonefish, 26 211 (96%) of which were annotated functionally with information from either sequence homology or protein signature searches.
Conclusions: We present the first genome of any anemonefish and demonstrate the value of low coverage (∼11×) long Nanopore read sequencing in improving both genome assembly contiguity and completeness. The near-complete assembly of the A. ocellaris genome will be an invaluable molecular resource for supporting a range of genetic, genomic, and phylogenetic studies specifically for clownfish and more generally for other related fish species of the family Pomacentridae.
One of the most iconic Australian fish is the Murray cod, Maccullochella peelii (Mitchell 1838), a freshwater species that can grow to ∼1.8 metres in length and live to age ≥48 years. The Murray cod is of a conservation concern as a result of strong population contractions, but it is also popular for recreational fishing and is of growing aquaculture interest. In this study, we report the whole genome sequence of the Murray cod to support ongoing population genetics, conservation, and management research, as well as to better understand the evolutionary ecology and history of the species. A draft Murray cod genome of 633 Mbp (N50 = 109 974bp; BUSCO and CEGMA completeness of 94.2% and 91.9%, respectively) with an estimated 148 Mbp of putative repetitive sequences was assembled from the combined sequencing data of 2 fish individuals with an identical maternal lineage; 47.2 Gb of Illumina HiSeq data and 804 Mb of Nanopore data were generated from the first individual while 23.2 Gb of Illumina MiSeq data were generated from the second individual. The inclusion of Nanopore reads for scaffolding followed by subsequent gap-closing using Illumina data led to a 29% reduction in the number of scaffolds and a 55% and 54% increase in the scaffold and contig N50, respectively. We also report the first transcriptome of Murray cod that was subsequently used to annotate the Murray cod genome, leading to the identification of 26 539 protein-coding genes. We present the whole genome of the Murray cod and anticipate this will be a catalyst for a range of genetic, genomic, and phylogenetic studies of the Murray cod and more generally other fish species of the Percichthydae family.
This study tested the oxygen consumption rates (OCR), energy, and thermal coefficient of juvenile Lutjanus malabaricus (60 fish, size: 4.53 ± 1.14 g) at four temperatures of 22, 26, 30 and 34 °C. During 30 days of experimental period 5 fish tank-1 were reared at four temperatures with three replicates in intermittent flow respirometers in a recirculatory system under laboratory conditions. As expected, oxygen consumption rates increased significantly (P
The contamination of seafood like narrow-barred Spanish mackerel (Scomberomorus commerson) fillets by potentially toxic elements (PTEs) has converted to worldwide health concerns. In this regard, the related citations regarding the concentration of PTEs in fillets of narrow-barred Spanish mackerel were collected through some of the international databases such as Scopus, Cochrane, PubMed, and Scientific Information Database (SID) up to 10 March 2020. The concentration of PTEs in fillets of narrow-barred Spanish mackerel fish was meta-analyzed and the health risk (non-carcinogenic risk) was estimated by the total target hazard quotient (TTHQ). The meta-analysis of data indicated that the rank order of PTEs in fillet of narrow-barred Spanish mackerel was Fe (10,853.29 μg/kg-ww) > Zn (4007.00 μg/kg-ww) > Cu (1005.66 μg/kg-ww) > total Cr (544.14 μg/kg-ww) > Mn (515.93 μg/kg-ww) > Ni (409.90 μg/kg-ww) > Pb (180.99 μg/kg-ww) > As (93.11 μg/kg-ww) > methyl Hg (66.60 μg/kg-ww) > Cd (66.03 μg/kg-ww). The rank order of health risk assessment based on the country by the aid of TTHQ for adult consumers was Malaysia (0.22251) > Philippines (0.21912) > Egypt (0.08684) > Taiwan (0.07430) > Bahrain (0.04893) > Iran (0.03528) > China (0.00620) > Pakistan (0.00316) > Yemen (0.00157) > India (0.00073). In addition, the rank order of health risk assessment based on the country by the aid of TTHQ for child consumers was Malaysia (1.03838) > Philippines (1.02257) > Egypt (0.40523) > Taiwan (0.34674) > Bahrain (0.22832) > Iran (0.16466) > China (0.02892) > Pakistan (0.01474) > Yemen (0.00731) > India (0.00340). Therefore, the children in Malaysia and the Philippines were at considerable non-carcinogenic risk. Hence, approaching the recommended control plans in order to decrease the non-carcinogenic risk associated with the ingestion of PTEs via the consumption of narrow-barred Spanish mackerel fish fillets is crucial.
The Labroides dimidiatus is one of the most traded marine ornamental fishes worldwide, yet not much is known about the microflora associated with this fish. This study is designed to investigate the bacteria composition associated with captive L. dimidiatus and its surrounding aquarium water. The fish and carriage water were obtained from well-known ornamental fish suppliers in Terengganu Malaysia. Bacteria present on the skin and in the stomach and the aquarium water were enumerated using culture-independent approaches and next-generation sequencing (NGS) technology. A total of 3,238,564 valid reads and 828 total operational taxonomic units (OTUs) were obtained from the three metagenomic libraries using NGS analysis. Of all the 15 phyla identified in this study, Proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria were the most prevalent in all samples. Also, 170 families belonging to 36 bacteria classes were identified. Although many of the bacteria families were common in the skin, gut, and aquarium water (39%), about 26% of the families were exclusive to the aquarium water alone. Therefore, any substantial change in the structure and abundance of microbiota (especially pathogenic bacteria) reported in this study may serve as an early sign for disease infection in the species under captivity. KEY POINTS: • Proteobacteria was the most dominant. • The microbiota was either shared or exclusively in samples.
Rhabdamia spilota Allen Kuiter 1994 (Apogonidae), a poorly known cardinalfish previously known only from the Philippines, Indonesia and the Red Sea, is redescribed on the basis of 70 specimens (20.9-61.2 mm standard length) (including types), from the Indo-West Pacific (Red Sea, Andaman Sea, Japan, South China Sea, the Philippines, Indonesia, New Caledonia, and Australia). Because most reports of the similar species R. gracilis (Bleeker 1856), following its original description, were based on misidentifications, R. gracilis is also redescribed (based on 98 Indo-West Pacific specimens from Seychelles, Maldives, Andaman Sea, Japan, Malaysia, Indonesia, New Caledonia, and Australia, 27.9-59.3 mm standard length); a lectotype is designated for it. Rhabdamia spilota differs from R. gracilis in having 27-33 (mode 30-31) developed gill rakers [vs. 22-27 (mode 24) in the latter], 27-33 (30) gill rakers including rudiments [vs. 23-27 (24-25)], a black stripe from the jaw tips to the anterior margin of the orbit (vs. black pigments only at snout and tip of lower jaw), 3-6 reddish brown to blackish blotches on the opercle and anterior of body (vs. blotches absent), and indistinct black pigment restricted to caudal fin outer margins (vs. pigment scattered over entire fin). Rhabdamia gracilis exhibits sexual dichromatism, female specimens larger than 41.3 mm SL having one or two black stripes on the lateral surface of the body; the stripes are absent in males and smaller females. No evidence of sexual dichromatism was found in R. spilota.
Both sexes of Brachiella malayensis n. sp. are described on the basis of specimens found in the nostrils of narrow-barred Spanish mackerel Scomberomorus commerson (Lacepède) collected off Besut, Malaysia. The female of this species closely resembles those of B. magna Kabata, 1968 and B. cybii Pillai, Prabha et Balaraman, 1982 but is distinguishable mainly by the body size and the proportions of the cephalosome, posterior processes and caudal rami. While examining the male, we noticed a systematic inconsistency in some lernaeopodid genera. The genus Brachiella Cuvier, 1830, represented by its type-species Brachiella thynni Cuvier, 1830, and two monotypic genera Charopinopsis Yamaguti, 1963 and Eobrachiella Ho et Do, 1984, represented by Charopinopsis quaternia (Wilson, 1935) and Eobrachiella elegans (Richiardi, 1880), respectively, share distinct synapomorphies in the embracing (vs. pinching) elongate male maxilliped and the female trunk with a pair of long, cylindrical ventroposterior processes (in addition to a pair of modified caudal rami), both of which are involved in their unique reproductive strategy. The latter two genera are herewith relegated to junior synonyms of Brachiella.
In the early 2000s, experimental rearing of spotted wolffish, Anarhichas minor, was started in Iceland. Health surveillance, carried out at regular intervals during the rearing period, revealed persistent and highly prevalent Kudoa infections of fish muscles which caused great financial losses due to post mortem myoliquefaction. In addition, during the traditional process of drying and smoking wild Atlantic lumpfish, Cyclopterus lumpus, the muscles from some fish almost completely disappear and the fish have to be discarded. To describe the etiological agent responsible for these conditions, spotted wolffish, Atlantic wolffish Anarhichas lupus, northern wolffish Anarhichas denticulatus and Atlantic lumpfish were caught off the Icelandic coast and examined for the presence of Kudoa. We describe a novel myxosporean, Kudoa islandica n. sp., using morphological and molecular data, and show with histopathology that it causes extensive myoliquefaction in three different wild fish hosts, which all are commercially valuable species in Iceland. Although some spore dimensions varied significantly between fish species, the molecular analyses showed that the same parasite was responsible for infection in all fish. The northern wolffish was not found to be infected. Although robustly placed in the Kudoa clade in phylogenetic analyses, K. islandica was phylogenetically distinct from other kudoids. A single myxosporean, K. islandica, is responsible for the infections in the somatic muscles of lumpfish and wolffish, causing extensive post mortem myoliquefaction. This myxosporean is likely to infect other fish species and it is important to study its life cycle in order to evaluate any threat to salmonid culture via the use of lumpfish as a biocontrol for sea lice.
We sequenced and assembled three whole mitogenome sequences of the commercially important snakeskin gourami Trichopodus pectoralis isolated from Malaysia (introduced), Viet Nam (native) and Thailand (native). The mitogenome length range from 16,397 to 16,420 bp. The final partitioned nucleotide alignment consists of 14,002 bp and supports the monophyly of the genus Trichopodus (95% ultrafast bootstrap support) with T. trichopterus forming a sister group with the members of T. pectoralis.
The caligid parasite Caligus coryphaenae has been reported in several marine fishes from Japan. In this study, a total of 223 wild yellowtail (Seriola quinqueradiata) were collected from Kagoshima fish market, separated into three groups according to total length (TL 70 cm), and examined for C. coryphaenae infection. A total of 518 specimens of C. coryphaenae were isolated giving an overall prevalence of infection and mean intensity of 24.2% and 4.7 ± 3.5, respectively. Body surface, fins and operculum of hosts TL > 50 cm were recorded as preferred sites for the attachment of C. coryphaenae. Of female specimens, 56.7% bore egg strings with various egg string lengths (7.1 ± 1.2 mm), carrying from 98 to 140 eggs (0.06 ± 0.005 mm in length) per each string. To our knowledge, this is the first report of C. coryphaenae infection from wild yellowtail in Japanese waters and globally. This parasite has not been reported in yellowtail culture to date, but the finding on wild specimens of yellowtail means that it poses a risk to their culture.
The mass mortality of cobia (Rachycentron canadum) within 2-3 days was reported by 3 private farms in Bukit Tambun, Pulau Pinang, in February and March 2007. Only cobia with body weights of 3-4 kg were affected. Most diseased cobia swam on the surface and displayed flashing behaviour. All samples were positive for viral nervous necrosis (VNN) with low to medium levels of infection. Infestations by leeches (Zeylanicobdella arugamensis), body monogeneans (Benedenia sp.) and copepods (Caligus sp.) were also found, but no pathogenic bacteria were isolated. All water quality parameters monitored were within optimal ranges for culturing cobia. The main causes of high mortality in cobia remain unclear during the study. However, we believe that the mass mortality of cobia could be probably due to VNN infection and that the rate of mortality will increase further when cobia are subjected to aquaculture-related stresses (e.g., limited space). Traditional cages with a size of 2 (length) × 2 (width) × 1 m (depth) should only be used for rearing cobia below 1 kg in weight given the species' natural behaviours. In addition, cobia fingerlings should be screened for VNN prior to stocking them in cages.
This study was conducted to determine the proximate composition and energetic value of selected marine fish and shellfish from the West Coast of Peninsular Malaysia. This study has included 20 species of fish (10 pelagic fish and 10 demersal fish) and 4 species of shellfish. For pelagic fish, Longtail shad (terubuk) was found to contain significantly lower moisture (59.31+0.00%), but significantly higher fat content (23.15+0.00%) compared to other fish species. For demersal fish, moisture and ash contents ranged between 74-82%, and 0.9-2.1%, respectively. Long-tailed butterfly ray contained the highest protein (22.22+1.24%) compared to other fish studied. The highest fat content of demersal fish was in Moonfish at 6.89+2.76%. For shellfish, prawn contained the highest protein (19.12+1.44%). The fat content of shellfish ranged between 1-2%. Oyster contained significantly higher carbohydrate at 6.45+0.00%, compared to other shellfish. Longtail shad contained the highest energetic value of 13.34 kJ g-1 of all samples. These values are useful references for consumers in order to choose fish and shellfish based on their nutritional contents.
The dragon millipede genus Desmoxytes s.l. is split into five genera, based on morphological characters and preliminary molecular phylogenetic analyses. The present article includes a review of Desmoxytes s.s., while future articles will deal with Hylomus Cook and Loomis, 1924 and three new genera which preliminarily are referred to as the 'acantherpestes', 'gigas', and 'spiny' groups. Diagnostic morphological characters of each group are discussed. Hylomus is resurrected as a valid genus and the following 33 species are assigned to it: H. asper (Attems, 1937), comb. n., H. cattienensis (Nguyen, Golovatch & Anichkin, 2005), comb. n., H. cervarius (Attems, 1953), comb. n., H. cornutus (Zhang & Li, 1982), comb. n., H. draco Cook & Loomis, 1924, stat. rev., H. enghoffi (Nguyen, Golovatch & Anichkin, 2005), comb. n., H. eupterygotus (Golovatch, Li, Liu & Geoffroy, 2012), comb. n., H. getuhensis (Liu, Golovatch & Tian, 2014), comb. n., H. grandis (Golovatch, VandenSpiegel & Semenyuk, 2016), comb. n., H. hostilis (Golovatch & Enghoff, 1994), comb. n., H. jeekeli (Golovatch & Enghoff, 1994), comb. n., H. lingulatus (Liu, Golovatch & Tian, 2014), comb. n., H. laticollis (Liu, Golovatch & Tian, 2016), comb. n., H. longispinus (Loksa, 1960), comb. n., H. lui (Golovatch, Li, Liu & Geoffroy, 2012), comb. n., H. minutuberculus (Zhang, 1986), comb. n., H. nodulosus (Liu, Golovatch & Tian, 2014), comb. n., H. parvulus (Liu, Golovatch & Tian, 2014), comb. n., H. phasmoides (Liu, Golovatch & Tian, 2016), comb. n., H. pilosus (Attems, 1937), comb. n., H. proximus (Nguyen, Golovatch & Anichkin, 2005), comb. n., H. rhinoceros (Likhitrakarn, Golovatch & Panha, 2015), comb. n., H. rhinoparvus (Likhitrakarn, Golovatch & Panha, 2015), comb. n., H. scolopendroides (Golovatch, Geoffroy & Mauriès, 2010), comb. n., H. scutigeroides (Golovatch, Geoffroy & Mauriès, 2010), comb. n., H. similis (Liu, Golovatch & Tian, 2016), comb. n., H. simplex (Golovatch, VandenSpiegel & Semenyuk, 2016), comb. n., H. simplipodus (Liu, Golovatch & Tian, 2016), comb. n., H. specialis (Nguyen, Golovatch & Anichkin, 2005), comb. n., H. spectabilis (Attems, 1937), comb. n., H. spinitergus (Liu, Golovatch & Tian, 2016), comb. n., H. spinissimus (Golovatch, Li, Liu & Geoffroy, 2012), comb. n. and H. variabilis (Liu, Golovatch & Tian, 2016), comb. n.Desmoxytes s.s. includes the following species: D. breviverpa Srisonchai, Enghoff & Panha, 2016; D. cervina (Pocock,1895); D. delfae (Jeekel, 1964); D. des Srisonchai, Enghoff & Panha, 2016; D. pinnasquali Srisonchai, Enghoff & Panha, 2016; D. planata (Pocock, 1895); D. purpurosea Enghoff, Sutcharit & Panha, 2007; D. takensis Srisonchai, Enghoff & Panha, 2016; D. taurina (Pocock, 1895); D. terae (Jeekel, 1964), all of which are re-described based mainly on type material. Two new synonyms are proposed: Desmoxytes pterygota Golovatch & Enghoff, 1994, syn. n. (= Desmoxytes cervina (Pocock, 1895)), Desmoxytes rubra Golovatch & Enghoff, 1994, syn. n. (= Desmoxytes delfae (Jeekel, 1964)). Six new species are described from Thailand: D. aurata Srisonchai, Enghoff & Panha, sp. n., D. corythosaurus Srisonchai, Enghoff & Panha, sp. n., D. euros Srisonchai, Enghoff & Panha, sp. n., D. flabella Srisonchai, Enghoff & Panha, sp. n., D. golovatchi Srisonchai, Enghoff & Panha, sp. n., D. octoconigera Srisonchai, Enghoff & Panha, sp. n., as well as one from Malaysia: D. perakensis Srisonchai, Enghoff & Panha, sp. n., and one from Myanmar: D. waepyanensis Srisonchai, Enghoff & Panha, sp. n. The species can mostly be easily distinguished by gonopod structure in combination with other external characters; some cases of particularly similar congeners are discussed. All species of Desmoxytes s.s. seem to be endemic to continental Southeast Asia (except the 'tramp' species D. planata). Some biological observations (relationship with mites, moulting) are recorded for the first time. Complete illustrations of external morphological characters, an identification key, and distribution maps of all species are provided.
The otoliths of sea bass (Lates calcarifer) were sampled from 23 locations in Peninsular Malaysia in order to analyse the Sr/Ca, Mg/Ca and Ba/Ca ratios. This study found that these elements/Ca ratios in sea bass otoliths were correlated with the salinity zone (thalassic series) compared with elements/Ca ratios in ambient water. The contradictory pattern of Sr/ Ca ratios and Ba/Ca ratios in otoliths was found according to salinity zone variations. Thus, oligohaline waters showed the highest Ba/Ca ratios while the highest Sr/Ca ratios were recorded for saline waters. The terrigenous enrichment of Ba in water also affects the Ba content in otoliths. The presence of Mg in otoliths acts as a rival to Ba, thus affecting Mg and Ba deposition. Meanwhile, Ba is the tracer for salinity fluctuations based on the partition coefficient (D) and it shows significant changes. However, the elements in otoliths were found to originate indirectly from the water column. The sequence of the elements/Ca ratios in otoliths and ambient water was Sr/Ca>Mg/Ca>Ba/Ca and Sr/Ca>Ba/Ca>Mg/ Ca, respectively.
Archerfishes Toxotes chatareus (Hamilton 1822) and Toxotes jaculatrix (Pallas 1767) inhabits mangrove estuaries, which are a critically important habitat as a spawning and nursery ground as well as the full life cycle of many fish species. In order to manage and conserve resilience fish species like the archerfish, we need to know some basic parameters about their biology and life history. Our research has focused on understanding the annual condition factors of these two species of archerfishes, as a complimentary to other several biological aspects of this two species that have been published. Our results indicated that, both species are in good conditions as the mean values of condition factors (Fulton condition factor K and Relative condition factor Kn) are greater than values of unity (K, Kn >1). The average K and Kn values of T. chatareus and T. jaculatrix were lowest in November and highest in September, indicates the spawning and recovery condition of the fishes. Condition factors measure overall fish population health that can be used by fisheries biologists and resource managers in proper management of fish resources in Malaysia waters.
A 60-day feeding trial was conducted to determine the effects of palm oil blended with oxidized and non-oxidized fish oil on growth performances, hematology, and non-specific immune response in juvenile Japanese sea bass, Lateolabrax japonicas. Japanese sea bass (1.73 ± 0.01 g) were fed seven experimental diets containing 100 g/kg of dietary lipid in forms of palm oil (10P), fish oil (10F), fish oil blended with palm oil at different ratios, 6:4 (6F4P) and 4:6 (4F6P), oxidized fish oil (10OF), and oxidized fish oil blended with palm oil at different ratios, 6:4 (6OF4P) and 4:6 (4OF6P). After the feeding trial, the following results were illustrated. No significant effects were observed in survival, feed conversion ratio, condition factor, and hematocrit after feeding with experimental diets for 60 days. The relatively higher specific growth rate and hematology were observed in 6F4P. Furthermore, both palm oil and oxidized fish oil acted as a negatively on serum lysozyme activity (P < 0.05). This study suggested that a ration of 6F4P is recommended as an innocuous ratio for Japanese sea bass. Furthermore, according to the present investigation, palm oil seems to have the ability to improve the protein efficiency when added to oxidized fish diets as well as a positive trend to the growth performance (P > 0.05).
There is very little information on the capacity of freshwater carnivorous fish to biosynthesize highly unsaturated fatty acids (HUFA). The striped snakehead fish (Channa striata) is a carnivorous species cultured inland of several Southeast Asian countries due to its pharmaceutical properties in wound healing enhancement. We described here the full-length cDNA cloning of a striped snakehead fatty acid desaturase (fads), which is responsible for desaturation of unsaturated fatty acids in the HUFA biosynthesis. Bioinformatics analysis reveals a protein coding region with length of 445 amino acids containing all characteristic features of desaturase enzyme, including a cytochrome b5-domain with the heme-binding motif, two transmembrane domains and three histidine-rich regions. The striped snakehead fads amino acid sequence shares high similarity with known fads of other teleosts. The mRNA expression of striped snakehead fads also showed an ontogenic-related increase in expression in 0-20 days after hatch larva. Using ISH, we localized the presence of fads in larva brain, liver and intestinal tissues.
Parasitic copepods infect a large number of fishes. In particular, the family Caligidae (Crustacea) comprises more than 450 species worldwide, some of which are harmful to cultured and wild fishes. Recently, we found a parasitic copepod on the host swordfish Xiphias gladius (Xiphidae) in the coastal waters of Parangipettai, southeast coast of India. It was identified as Gloiopotes huttoni by its morphological features, and it belongs to the family Caligidae (order Siphonostomatoida). The swordfish was highly infected with about 72 caligids on a single host. Both male and female forms of G. huttoni were colonized on the beak of the swordfish and distributed mostly along the upper and lower jaws of the host, with 42 and 30 copepod parasites, respectively. Most of the copepods were ovigerous, and more females than males were recorded. It is the first record of the occurrence of G. huttoni in India.
This study provides a first description of the phylogeographic patterns and evolutionary history of two species of the mudskipper genus Periophthalmus. These amphibious gobies are distributed throughout the whole Indo-Pacific region and Atlantic coast of Africa, in peritidal habitats of soft-bottom coastal ecosystems. Three sequence datasets of two widely distributed species, Periophthalmus argentilineatus and P. kalolo, were obtained by amplifying and sequencing two mtDNA markers (D-loop and 16S rDNA) and the nDNA rag1 region. The three datasets were then used to perform phylogeographic, demographic and population genetic analyses. Our results indicate that tectonic events and past climatic oscillations strongly contributed to shape present genetic differentiation, phylogeographic and demographic patterns. We found support for the monophyly of P. kalolo, and only shallow genetic differentiation between East-African and Indo-Malayan populations of this species. However, our collections of the morphospecies P. argentilineatus include three molecularly distinct lineages, one of them more closely related to P. kalolo. The presence of Miocenic timings for the most recent common ancestors of some of these morphologically similar clades, suggests the presence of strong stabilising selection in mudskippers' habitats. At population level, demographic analyses and palaeoecological records of mangrove ecosystems suggest that Pleistocene bottlenecks and expansion plus secondary contact events of the studied species were associated with recurrent sea transgressions during interglacials, and sea regressions or stable regimes during glacials, respectively.
The purpose of this study was to characterize major allergens of Indian scad (Decapterus russelli) which is among the most commonly consumed fish in Malaysia. Raw and cooked extracts of the fish were prepared. Protein profiles and IgE binding patterns were produced by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting using sera from subjects with fish allergy. The major allergens of the fish were then identified by two-dimensional electrophoresis (2-DE), followed by mass spectrometry of the peptide digests. The SDS-PAGE of the raw extract revealed 27 protein fractions over a wide molecular weight range, while the cooked extract demonstrated only six protein fractions. The 1-DE immunoblotting detected 14 IgE-binding proteins, with a molecular weight range from 90 to < 6.5 kDa. Three protein fractions with molecular weights of approximately 51, 46 and 12 kDa were identified as the major allergens of this fish. The approximately 12 kDa band was a heat-resistant protein while the approximately 51 and 46 kDa proteins were sensitive to heat. The 2-DE gel profile of the raw extract demonstrated > 100 distinct protein spots and immunoblotting detected at least 10 different major IgE reactive spots with molecular masses as expected and isoelectric point (pI) values ranging from 4.0 to 7.0. A comparison of the major allergenic spot sequences of the 12 kDa proteins with known protein sequences in databases revealed extensive similarity with fish parvalbumin. In conclusion, this study demonstrated that a parvalbumin which is similar to Gad c 1 is the major allergen of Indian scad. Interestingly, we also detected heat-sensitive proteins as major allergenic components in our fish allergy patients.