This study was conducted to determine the cholesterol and alpha-tocopherol contents of 20 marine fish and four other seafood from the Straits of Malacca. Cholesterol and alphatocopherol contents of the fish and other seafood were determined using high-performance liquid chromatography. The results showed that most of the fish contained low amounts of cholesterol, except sixbar grouper (Epinephelus fasciatus), long-tailed butterfly ray (Gymnura sp.), yellowstripe scad (Selaroides leptolepis), cuttlefish (Sepia officinalis), large-scale tongue sole (Cynoglossus arel), and longtail shad (Hilsa macrura) that contained high amounts of cholesterol (119.39-353.97 mg/100 g wet samples). Indian mackerel (Rastrelliger kanagurta), giant seaperch (Lates calcarifer), prawn (Metapenaeus affinis), and moonfish (Trachinotus blochii) had high alpha-tocopherol contents (462-989 μg/100 g wet sample). Regular consumption of fish and other seafood is highly recommended partly due to the high alphatocopherol content. Due to the high cholesterol in certain types of fish, consumption of the fish fillets of sixbar grouper, long-tailed butterfly ray, yellowstripe scad, cuttlefish, and large scale tongue sole should be < 100 g per day and < 50 per day for longtail shad. Validation of the analytical method also showed a high accuracy and reproducibility of the HPLC method.
Freshwater fish has been studied and reported numerously. However, little attention has been made and limited studies available on local marine fish in Malaysia. Thus, in this study, concentrations of heavy metals (Cd, Cr, Pb and Cu) were studied in four major local marine fish Megalaspis cordyla (hardtail scad), Rastrelliger kanagurta (Indian mackerel), Selaroides leptolepis (yellowstripe scad) and Sardinella fimbriata (fringescale sardinella). The study was also intended to estimate potential health risk assessment from these heavy metals to the consumption of fish and assess maximum allowable fish consumption rate. The range of heavy metal concentrations were 0.053-0.096 mg/kg for Cd, 1.16-2.34 mg/kg for Cr, 8.34-12.44 mg/kg for Pb and 1.40-3.21 mg/kg for Cu in four major self-caught saltwater fish. Heavy metal levels of Cd and Cu in the local marine fish from Port Dickson are below the limit enforced by Food Regulations (1985) while the levels of Cr and Pb have exceeded the limit. Potential health risks associated with Cd, Cr, Cu and Pb were assessed based on target hazard quotients. HQ values calculated for Cd, Cr and Cu were less than 1, thus indicate that no adverse effects while HQ values for Pb exceeded 1 for all the fish species assessed with the exception of Megalaspis spp and Sardinella sppa. Cr was the highest while Pb concentrations were the lowest in all the studied fish samples for maximum allowable fish consumption rate. A long term monitoring program is crucial to be done in coastal areas with high consumption of local marine fish along Port Dickson to obtain real consumption rates and other cofounders factors in local population.
A study on the variation of leaf venation patterns was conducted on 21 taxa of the genus Ficus in Peninsular Malaysia. The results showed the existence of eight leaf venation patterns based on veinlets, the ultimate marginal and areolar venation. The majority of species, such as F. annulata, F. benghalensis, F. benjamina, F. deltoidea var. angustifolia, F. deltoidea var. kunstleri, F. depressa, F. elastica, F. hispida, F. microcarpa, F. religiosa, F. tinctoria, F. ucinata and F. vasculosa, show tri-veinlets. The others exhibit the following: bi-veinlets in F. aurata and F. heteropleura; uni-veinlets in F. lepicarpa, F. schwarzii and F. superba; and simple veinlets in F. aurantiacea and F. fulva. F. sagittata presents no veinlets for areolar venation. The presence of tracheid or swollen veins at the centre of the lamina and the presence of cystolith cells and trichomes are common anatomical characteristics that could assist in group classification of the studied species. Variations in leaf venation patterns are not only valuable in identifying a taxon group, but can also be used to differentiate between species in the genus Ficus.
In this study, we reported a molecular characterization of a novel proto-type galectin-1 from the striped murrel Channa striatus (named as CsGal-1). The full length CsGal-1 was identified from an established striped murrel cDNA library and further we confirmed the sequence by cloning. The complete cDNA sequence of CsGal-1 is 590 base pairs (bp) in length and its coding region encoded a poly peptide of 135 amino acids. The polypeptide contains a galactoside binding lectin domain at 4-135. The domain carries a sugar binding site at 45-74 along with its signatures (H(45)-X-Asn(47)-X-Arg(49) and Trp(69)-X-X-Glu(72)-X-Arg(74)). CsGal-1 shares a highly conserved carbohydrate recognition domain (CRD) with galectin-1 from other proto-type galectin of teleosts. The mRNA expressions of CsGal-1 in healthy and various immune stimulants including Aphanomyces invadans, Aeromonas hydrophila, Escherchia coli lipopolysaccharide and poly I:C injected tissues of C. striatus were examined using qRT-PCR. CsGal-1 mRNA is highly expressed in kidney and is up-regulated with different immune stimulants at various time points. To understand its biological activity, the coding region of CsGal-1 gene was expressed in an E. coli BL21 (DE3) cloning system and its recombinant protein was purified. The recombinant CsGal-1 protein was agglutinated with mouse erythrocytes at a concentration of 4μg/mL in a calcium independent manner. CsGal-1 activity was inhibited by d-galactose at 25mM(-1) and d-glucose and d-fructose at 100mM(-1). The results of microbial binding assay showed that the recombinant CsGal-1 protein agglutinated only with the Gram-negative bacteria. Interestingly, we observed no agglutination against Gram-positive bacteria. Overall, the study showed that CsGal-1 is an important immune gene involved in the recognition and elimination of pathogens in C. striatus.
In February 2013, forty-seven Notched threadfin bream, the Nemipterus peronii, were sampled from the eastern coastal waters of the South China Sea. The concentration of various elements, namely cadmium (Cd), chromium (Cr), copper (Cu), mercury (Hg), strontium (Sr), manganese (Mn), selenium (Se), Lead (Pb), nickel (Ni), aluminum (Al), arsenic (As), iron (Fe), and Zinc (Zn) were analyzed in the liver, muscle, and kidney organs of the host, as well as in their parasites Hysterothalycium reliquens (nematode) and the Paraphilometroides nemipteri (nematode), using inductively coupled plasma mass spectrometry (ICP-MS). The former group of parasites showed highest accumulation capacity for Cr, Cu, Fe, Mn, Se, Ni, and Zn while the latter group had high accumulation potential of As, Hg, Cd, Al, Pb, and Sr. The divergence in heavy-metal accumulation profiles of both nematodes is linked with the specificity of microhabitats, cuticle morphology, and interspecific competition. The outcome of this study indicates that both parasite models can be used for biomonitoring of metal pollution in marine ecosystems.
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.
CXCR3 is a CXC chemokine receptor 3 which binds to CXC ligand 4 (CXCL4), 9, 10 and 11. CXC chemokine receptor 3a (CXCR3a) is one of the splice variants of CXCR3. It plays crucial role in defense and other physiological processes. In this study, we report the molecular cloning, characterization and gene expression of CXCR3a from striped murrel Channa striatus (Cs). The full length CsCXCR3a cDNA sequence was obtained from the constructed cDNA library of striped murrel by cloning and sequencing using an internal sequencing primer. The full length sequence is 1425 nucleotides in length including an open reading frame of 1086 nucleotides which is encoded with a polypeptide of 361 amino acids (mol. wt. 40 kDa). CsCXCR3a domain analysis showed that the protein contains a G protein coupled receptor between 55 and 305 along with its family signature at 129-145. The transmembrane prediction analysis showed that CsCXCR3a protein contains 7 transmembrane helical regions at 34-65, 80-106, 113-146, 154-181, 208-242, 249-278 and 284-308. The 'DRY' motif from CsCXCR3a protein sequence at (140)Asp-(141)Arg-(142)Tyr which is responsible for G-protein binding is also highly conserved with CXCR3 from other species. Phylogenetic tree showed that the CXC chemokine receptors 3, 4, 5 and 6, each formed a separate clade, but 1 and 2 were clustered together, which may be due to the high similarity between these receptors. The predicted 3D structure revealed cysteine residues, which are responsible for 'CXC' motif at 116 and 198. The CsCXR3a transcript was found to be high in kidney, further its expression was up-regulated by sodium nitrite acute toxicity exposure, fungal, bacterial and poly I:C challenges. Overall, these results supported the active involvement of CsCXCR3a in inflammatory process of striped murrel during infection. However, further study is necessary to explore the striped murrel chemokine signaling pathways and their roles in defense system.
The gram-negative bacterium, Vibrio alginolyticus, has frequently been identified as the pathogen responsible for the infectious disease called vibriosis. This disease is one of the major challenges facing brown-marbled grouper aquaculture, causing fish farmers globally to suffer substantial economic losses. The objective of this study was to investigate the proteins involved in the immune response of brown-marbled grouper fingerlings during their initial encounter with pathogenic organisms. To achieve this objective, a challenge experiment was performed, in which healthy brown-marbled grouper fingerlings were divided into two groups. Fish in the treated group were subjected to intraperitoneal injection with an infectious dose of V. alginolyticus suspended in phosphate-buffered saline (PBS), and those in the control group were injected with an equal volume of PBS. Blood samples were collected from a replicate number of fish from both groups at 4 h post-challenge and analysed for immune response-related serum proteins via two-dimensional gel electrophoresis. The results showed that 14 protein spots were altered between the treated and control groups; these protein spots were further analysed to determine the identity of each protein via MALDI-TOF/TOF. Among the altered proteins, three were clearly overexpressed in the treated group compared with the control; these were identified as putative apolipoprotein A-I, natural killer cell enhancement factor and lysozyme g. Based on these results, these three highly expressed proteins participate in immune response-related reactions during the initial exposure (4 h) of brown-marbled grouper fingerling to V. alginolyticus infection.
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 high prevalence (80-100%) of the marine leech Zeylanicobdella arugamensis (De Silva) on cage-cultured Asian sea bass Lates calcarifer (Bloch) led us to investigate the percentage of juvenile leeches hatched from deposited cocoons, survival of juvenile and adult marine leeches at different salinity and temperature. The results showed that the hatching percentage of juvenile leeches was highest at salinity of 30 ppt (32.5 ± 2.8%) followed by 20 ppt (18.0 ± 4.3%) and 10 ppt (12.1 ± 1.4%), respectively. It was found that the adult and juvenile leeches could live up to an average range of 4-7 days at salinity ranging from 10 to 40 ppt. The juvenile leeches were able to hatch at temperature ranging from 25 to 35 °C but unable to hatch at 40 °C. The survival period of adult and juvenile leeches ranged from 11 to 16 days at 25 °C, which was comparatively longer than 5-13 days and 10 h--5 days at 27-30 °C and 35-40 °C, respectively. The study provided the information on the physical parameters of salinity and temperature which are most optimal for the marine leech Z. arugamensis to propagate.
The authors studied the myxosporean infection of wild gobiid fishes (Perciformes: Gobioidei) in the Merang Estuary of Terengganu, Malaysia, and described Myxobolus ophiocarae sp. n. in Ophiocara porocephala. Several myxosporean plasmodia were found intralamellarly within the gill filaments. The spores differed from those of other Myxobolus species previously recorded on gobiid fishes. They were round in valvular view and lens-shaped in sutural view, and had two equal-sized, pyriform polar capsules with polar filaments having six to seven turns. The spores measured 10.34 × 8.79 × 4.53 μm. The 18S rDNA sequence of M. ophiocarae sp. n., based on a contiguous sequence of 1,789 base pairs, differed from any other Myxobolus spp. in GenBank. Phylogenetic analysis of the 18S rDNA gene revealed that this species showed the closest similarity to Myxobolus nagaraensis, Myxobolus lentisuturalis, and Myxobolus cultus.
We report here the complete genome sequence of C. neteri SSMD04, a strain isolated from pickled mackerel sashimi, sequenced by third-generation sequencing technology. To the best of our knowledge, this is the first documentation that reports the complete genome of Cedecea neteri.
This study aims to optimise the operating conditions for the supercritical fluid extraction (SFE) of toxic elements from fish oil. The SFE operating parameters of pressure, temperature, CO2 flow rate and extraction time were optimised using a central composite design (CCD) of response surface methodology (RSM). High coefficients of determination (R²) (0.897-0.988) for the predicted response surface models confirmed a satisfactory adjustment of the polynomial regression models with the operation conditions. The results showed that the linear and quadratic terms of pressure and temperature were the most significant (p < 0.05) variables affecting the overall responses. The optimum conditions for the simultaneous elimination of toxic elements comprised a pressure of 61 MPa, a temperature of 39.8ºC, a CO₂ flow rate of 3.7 ml min⁻¹ and an extraction time of 4 h. These optimised SFE conditions were able to produce fish oil with the contents of lead, cadmium, arsenic and mercury reduced by up to 98.3%, 96.1%, 94.9% and 93.7%, respectively. The fish oil extracted under the optimised SFE operating conditions was of good quality in terms of its fatty acid constituents.
The aims of this study were to examine the effect of salts (CaCl2, CaSO4 and MgSO4) on the rheological and thermal properties of gelatin extracted from the skins of tropical fishes, sin croaker (Johnius dussumeiri) and shortfin scad (Decapterus macrosoma). It was found that the melting temperatures of fish skin gelatins were increased by 1.5 times as compared to bovine gelatin which was only increased by 0.5 times after holding for 2 h at 5°C. The storage (G’) and loss (G”) modulus of fish skin gelatins were improved with the addition of calcium sulphate (CaSO4) and magnesium sulphate (MgSO4), respectively. However, the storage (G’) and loss (G”) modulus of gelatin solutions were decreased with the addition of calcium chloride (CaCl2). Magnesium sulphate (MgSO4) was found to be an effective salt to improve the bloom value, elastic and viscous moduli of the fish skin gelatin. This study showed that shortfin scad skin gelatin with salt addition possessed better thermal and rheological properties than sin croaker gelatin.
Channa striatus (“haruan”) fish destined for fillet preparation was subjected to two freezing treatments, freezing with distilled water (FW) or freezing directly without distilled water (DF). Fish that was freshly processed without freezing served as control (C). Fillet yield (%) was in the range 33.8% to 35.3% and the highest yield was recorded in FW samples. Whole Fillet Powder (WFP) was prepared from the fillets through low temperature vacuum oven drying (50°C) and its composition and physicochemical properties were assessed. There was no significant difference in moisture and protein contents of all samples (p > 0.05). All WFP were generally dark in colour with whiteness indices ranging from 55.23 - 63.98. The redness (a*) values were 4.33, 11.12, 8.83 whilst the yellowness (b*) were 19.31, 23.04, 21.20 for C, WFP-FW and WFP-DF respectively. WFPs were generally high in histidine, arginine, threonine and tyrosine when compared to egg whites and these (except histidine) and other amino acids (serine, glycine, methionine and phenylalanine) were significantly higher (p < 0.05) in WFP-FW compared to other samples. Overall, freezing treatments affected the composition and physicochemical properties of WFPs.
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.
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.
Culturing fishes in marine cages is a rapidly developing area of marine aquaculture. The Asian seabass Lates calcarifer (Bloch) is a fast growing good quality fish that is readily cultured in intensive systems in the South Asian region and in Malaysia in particular. Although several papers have been published to date on viral, bacterial, parasitic and fungal organisms causing diseases in the Asian seabass, the occurrence of a coccidian infection in this species has only recently been recorded. We collected sporulated and unsporulated oöcysts of a new species of Goussia Labbé, 1986, from the mucus covering the epithelium of the intestine of L. calcarifer. This paper provides a description of Goussia kuehae n. sp. Sporulated oöcysts of this species are ellipsoidal, 37-40 μm in length and 28-30 μm in width. The ellipsoidal sporocysts are relatively small, 15.2-17 × 5.7-8 μm, and located loosely in the oöcyst. There are residual bodies both in the oöcysts and the sporocysts. Goussia kuehae n. sp. differs from all known species of Goussia in the large size of the oöcysts and in having two types of oöcyst residuum.
A new proteomics technology has been implemented to study the protein repertoires of developing oocytes of giant grouper (Epinephelus lanceolatus). Knowledge of the chemical composition and physiochemical properties of vitellogenin (Vtg) is necessary to interpret the functional and biological properties attributed during ovulation. Vtg, as a biomarker indicator in sex determination, has been analyzed to determine the sex and maturational status of fish in the absence of the gonad tissue. A male giant grouper was induced by 2 mg/kg of 17ß-estradiol (E2), and blood was sampled at days 0, 1, 3, 5, and 10. SDS-PAGE 1D electrophoresis was used to analyze Vtg protein, and Vtg identification was done with 4800 Plus MALDI TOF/TOF™ mass spectrophotometer (Applied Biosystems/MDS SCIEX, USA). Meanwhile, MS/MS de novo sequencing identified the proteins by matching sequences of tryptic peptides to the known sequences of other species. Vtg was confirmed by MASCOT at 95% significant level, and molecular mass was 187 kDa. Protein resolved on SDS-PAGE as a double band of approximately the same mass as determined with MALDI-TOF. The N-terminal sequences and identification of Vtg were also determined. The potential of using MS methods to understand the structure and function of Vtg is discussed.
Histamine, putrescine cadaverine and cis-urocanic acid (UCA) have all been implicated or suggested in scombroid fish poisoning. However, there is little information on UCA especially during storage. Changes in their contents during storage of whole Indian mackerel at 0, 3±1, 10±1 for up to 15 days and 23±2°C for up to 2 days were monitored. Fresh muscles contained 14.83 mg/kg trans-UCA, 2.23 mg/kg cis-UCA and 1.86 mg/kg cadaverine. Histamine and putrescine were not detected. After 15 days at 0 and 3°C, trans-UCA content increased to 52.83 and 189.51 mg/kg, respectively, and decreased to <2 mg/kg at the other two temperatures. Storage at 10°C also resulted in an increase in trans-UCA after 3 days, only to decrease after 6 days. The concentration of cis-UCA increased nearly 13-fold after 15 days at 0 and 3°C, decreased at 10°C and remained unchanged at 23°C. Histamine, putrescine and cadaverine levels increased significantly (P value<0.05) at all temperatures especially at 23°C.