Siti Shapor Siraj

Use of different soft tissues of Perna viridis a s biomonitors of bioavailability and contamination by heavy metals (Cd, Cu, Fe, Pb, Ni, and Zn) in a semi-enclosed intertidal water, the Johore Straits

Green-lipped mussels, Perna viridis, were collected from the eastern and western parts of the Johore Straits in September 2004 and January 2005. Based on the heavy metal concentrations in the different soft tissues (gonad, foot, mantle, gills, muscle, and remaining soft tissues) of these mussel samples, the eastern part of the Johore Straits (which is divided into two portions by a causeway), recorded higher levels of bioavailability and contamination by Cd, Cu, Fe, Ni, and Zn when compared to the western part, while Kg. Pasir Puteh in the eastern part was found to record the highest bioavailability and contamination by heavy metals. The use of different soft tissues of P. viridis as biomonitors of bioavailability and contamination by Cd, Cu, Fe, Pb, Ni, and Zn in the semi-enclosed Johore Straits is proposed, since erroneous results due to spawning and the problem of defecation before dissection could be overcome. Hence, a more accurate interpretation of the bioavailability and contamination by heavy metals in coastal waters could be obtained. To our knowledge, this is the most detailed study on the bioavailability and contamination of heavy metals in the Johore Straits on the Malaysian side of the waterway carried out by using the different soft tissues and metal distribution based on the Mussel Watch approach.

Identification and characterization of Malaysian river catfish, Mystus nemurus (C&V): RAPD and AFLP analysis.

This work represents the first application of the amplified fragment length polymorphism (AFLP) technique and the random amplified polymorphic DNA (RAPD) technique in the study of genetic variation within and among five geographical populations of M. nemurus. Four AFLP primer combinations and nine RAPD primers detected a total of 158 and 42 polymorphic markers, respectively. The results of AFLP and RAPD analysis provide similar conclusions as far as the population clustering analysis is concerned. The Sarawak population, which is located on Borneo Island, clustered by itself and was thus isolated from the rest of the populations located in Peninsular Malaysia. Both marker systems revealed high genetic variability within the Universiti Putra Malaysia (UPM) and Sarawak populations. Three subgroups each from the Kedah, Perak, and Sarawak populations were detected by AFLP but not by RAPD. Unique AFLP fingerprints were also observed in some unusual genotypes sampled in Sarawak. This indicates that AFLP may be a more efficient marker system than RAPD for identifying genotypes within populations.

Biochemical Polymorphism in Yellow Catfish, Mystus nemurus (C&V), from Thailand

Yellow catfish, Mystus nemurus (Cuv. & Val.), is becoming one of the major freshwater species farmed by aquaculturists in Southeast Asia. It was of interest to examine levels of genetic subpopulation differentiation among samples of this species obtained from parts of its range, as well as to compare the genetics of wild and hatchery-bred fish. Horizontal starch gel electrophoresis and histochemical staining techniques were used to examine genetic variation within and among eight wild and one hatchery populations of M. nemurus from northern, northeastern, central and southern Thailand. Four tissues (heart, liver, kidney, and muscle) from individual specimens were used to analyze variations at 23 protein-coding loci. Fifteen of the 23 loci examined (65.22%), namely, ACP*, AAT-1*, EST-1*, EST-2*, GPI*, IDH-1*, IDH-2*, MDH-1*, MDH-2*, MDH-3*, ME*, PGM*, 6PGD*, SOD*, and HB*,were polymorphic at the 0.95 level. Observed heterozygosities ranged from 0.041 to 0.111, with an average of 0.068 ± 0.028. Genetic distances ranged from 0.005 to 0.164. The greatest genetic distance was found between the Chainat and the Suratthani populations (0.164), a level indicative of subspecific differentiation in M. nemurus from within Thailand.

Association of annual hormonal profile with gonad maturity of mahseer (Tor tambroides) in captivity.

Annual gonad hormonal profile of wild, matured mahseer (29 males and 23 female) averaging in weight between 0.95±0.26 and 1.19±0.23 kg for males and females, respectively, were investigated from November 2007 to November 2008 using enzyme-linked immunosorbent assay (ELISA) technique. Blood was collected from caudal vein, monthly and plasma separation by centrifugation was done to measure reproductive hormones: 17β-estradiol (E(2)), testosterone (T), and 11-keto-testosterone (11KT). Gonads were sampled for histology processing to observe their maturity. Highest T level in females and males was recorded at 0.22±0.016 and 0.88±0.014 ng/ml, respectively. The 11KT showed several peaks and the highest value was noted at 0.7±0.018 ng/ml in November 2008. The female E(2) initially was at 1.48±0.16 ng/ml and significantly increased (P<0.05) to 1.53±0.39 ng/ml in November 2008. Ovaries were laden with oocytes in several stages in all the samples while testes gonad showed a high level of spermatids throughout the year. Changes in plasma level of the gonadal hormones were correlated with the ovarian and testes maturities. In conclusion, the study suggests that mahseer can be categorized as asynchronized and multiple spawner. The information gathered is important for appropriate breeding and conservation programs of the Malaysian mahseer.

Interpopulation crosses, inheritance study, and genetic variability in the brown planthopper complex, Nilaparvata lugens (Homoptera: Delphacidae).

Studies on hybridization, inheritance, and population genetics of brown planthoppers that infest rice and weeds were undertaken using starch gel electrophoresis to determine whether the weed-infesting population represents a biological race or a species. F1 and F2 generations were produced by crosses between parental insects from the two populations with little indication of hybrid sterility. Gpi, Mdh, and Idh loci were inherited in a simple Mendelian fashion in families of two sympatric populations. Sixteen populations of Nilaparvata spp. from eight locations were collected. The Mdh, Idh, Pgm, Gpi, 6Pgd, and Acp loci were polymorphic. The N. lugens of rice with high esterase activity were clustered into a group and characterized by the presence of alleles Gpi110 and Gpi120, whereas N. lugens from weeds with low esterase activity were clustered into another group and characterized by Gpi100 and Gpi90. There was a lack of heterozygotes between the common alleles of the two populations. This means that the two groups of individuals belong to different gene pools.

Evidence for a higher number of species of Odontotermes (Isoptera) than currently known from Peninsular Malaysia from mitochondrial DNA phylogenies.

Termites of the genus Odontotermes are important decomposers in the Old World tropics and are sometimes important pests of crops, timber and trees. The species within the genus often have overlapping size ranges and are difficult to differentiate based on morphology. As a result, the taxonomy of Odontotermes in Peninsular Malaysia has not been adequately worked out. In this study, we examined the phylogeny of 40 samples of Odontotermes from Peninsular Malaysia using two mitochondrial DNA regions, that is, the 16S ribosomal RNA and cytochrome oxidase subunit I genes, to aid in elucidating the number of species in the peninsula. Phylogenies were reconstructed from the individual gene and combined gene data sets using parsimony and likelihood criteria. The phylogenies supported the presence of up to eleven species in Peninsular Malaysia, which were identified as O. escherichi, O. hainanensis, O. javanicus, O. longignathus, O. malaccensis, O. oblongatus, O. paraoblongatus, O. sarawakensis, and three possibly new species. Additionally, some of our taxa are thought to comprise a complex of two or more species. The number of species found in this study using DNA methods was more than the initial nine species thought to occur in Peninsular Malaysia. The support values for the clades and morphology of the soldiers provided further evidence for the existence of eleven or more species. Higher resolution genetic markers such as microsatellites would be required to confirm the presence of cryptic species in some taxa.

Biochemical studies on malathion resistance, inheritance and association of carboxylesterase activity in brown planthopper, Nilaparvata lugens complex in Peninsular Malaysia

Abstract  Two sympatric populations of brown planthopper (BPH), one from rice and the other from Leersia hexandra were collected from each of five locations in Malaysia. All the tested malathion‐resistant individuals of the rice BPH population and F1 generation (cross between malathion‐resistant [usually caught on rice] and malathion‐susceptible [usually caught on Leersia]) showed high esterase activity, while all malathion‐susceptible individuals on L. hexandra showed low esterase activity. In the F2 generation, all the individuals tested against malathion were approximately 75% resistant and 25% susceptible and the inheritance pattern of esterase activity (high and low esterase activity) segregated in the same manner to a 3: 1 ratio. This confirms that resistance to malathion is mono‐factorial and inheritance pattern of esterase activity is also linked to malathion resistance. Carboxylesterase or total esterase activity in BPH is inherited in a simple Mendelian fashion that is encoded by a single dominant gene. For the total esterase assay, average esterase activity levels in the rice‐infesting population ranged from 17.64 to 19.37 nmoles 1‐napthol/mg protein while that in the Leersia‐infesting population ranged from 5.29 to 6.11 nmoles 1‐napthol/mg protein. In terms of esterase activity, the two sympatric Nilaparvata lugens populations separated into two distinct groups. Results based on the tube color intensity test showed 96% and 98% resistant and susceptible individuals were present in the rice‐ and Leersia‐infesting populations, respectively. In a filter paper test, the rice‐infesting population had 94% with high esterase activity while the Leersia‐infesting population had 96% with low esterase activity.

Ovarian development of a river catfish Hemibagrus nemurus (Valenciennes, 1840) in captivity.

Hemibagrus nemurus is a riverine catfish with high economic and nutritive values. Investigations on ovarian development of this fish were carried out to determine the mode of ovarian development and describe the oocyte developmental stages. Histological studies were done on ovaries using light microscopy and scanning electron microscopy. Fish were sampled monthly for a period of six months (August 2009 to January 2010). The mean oocyte diameter (OD) ranged from 871 ± 161.41 µm to 1,167 ± 26.77 µm and the highest OD was in November. Oocyte size-frequency distribution showed a polymodal distribution. The mean gonadosomatic index (GSI) ranged from 1.14 ± 0.87% to 7.06 ± 1.40% and highest GSI was in November. The ovaries exhibited three phases of oocyte growth, which were primary growth, secondary growth and maturation phases. Based on histological criteria, the oocyte developmental stages were divided into seven stages as chromatin nucleolar, early perinucleolar, late perinucleolar, cortical alveolar, vitellogenesis, mature oocyte and germinal vesicle migration stages. All the seven stages of oocyte development were observed in the ovaries. Oogonia were always present throughout the developmental stages. The ovaries had more than two stages of oocyte development. This is the first report on the mode of ovarian development of H. nemurus. These findings indicated that H. nemurus has asynchronous mode of ovarian development and is capable of spawning several times in a year under favourable conditions.

Development of Microsatellite Markers from an Enriched Genomic Library for the Genetic Analysis of the Malaysian Giant Freshwater Prawn, Macrobrachium rosenbergii

Prawns of the genus Macrobrachium Bate, 1868 (Crustacea: Palaemonidae), are a highly diverse, abundant, and widespread group of decapod crustaceans found in circumtropical marine, estuarine, and freshwater areas (de Bruyn et al. 2004). The giant freshwater prawn Macrobrachium rosenbergii is the largest species in the genus and is the most important culture species. It is being cultured in commercial quantities in many parts of the world, including Hawaii, Honduras, Mauritius, Taiwan, Thailand, Philippines, Costa Rica, Israel, Malaysia, and Mexico (New 2002). Overexploitation of M. rosenbergii, however, will eventually lead to the loss of genetic diversity.

Isolation and characterization of microsatellite loci in the Malaysian giant freshwater prawn, Macrobrachium rosenbergii

Eight single locus microsatellite markers were developed to characterize the Malaysian giant freshwater prawn, Macrobrachium rosenbergii. These microsatellites were isolated from an enriched genomic library contained by using a 5′‐anchored polymerase chain reaction technique. Primers were designed to flank the repeat sequences and subsequently used to characterize 30 unrelated individuals of the giant freshwater prawn. The polymerase chain reaction amplification products of these eight microsatellite loci were polymorphic with the number of alleles ranging from two to 10 alleles per locus while the levels of heterozygosity ranged from 0.6333 to 0.8667.