Production of Recombinant Envelope Proteins of Newcastle Disease Virus in Escherichia Coli and Analysis of Their Immunological Properties

Newcastle disease virus (NDV) is the causative agent of the Newcastle disease (ND) thatremains as a major threat to the world poultry industry. The virus belongs to the family Paramyxoviridae and genus Avulavirus, infects more than 236 avian species, and causes up to 100% morbidity and mortality in susceptible birds. The viral envelope proteins, haemagglutinin-neuraminidase (HN) and fusion (F) proteins have been shown to play key roles in triggering the host immune responses. In order to study the immunological properties of the recombinant HN and F proteins, the HN and F genes of the Malaysian viscerotropic-velogenic NDV strain AF2240 were obtained through reverse transcription-polymerase chain reaction (RT-PCR) and cloned into the Pichia pastoris, Saccharomyces cerevisiae and Escherichia coli expression vectors. A number of eight recombinant plasmids were constructed, namely pPICZaA/HN and pPICZaA/F (P. pastoris system), pYES2a/HN and pYES2a/F (S. cerevisiae system),and pRSETA/HN, pRSETB/F, pET-43.1a/HN and pET-43.1a/F (E. coli system). The recombinant plasmids were used to transform respective host cells, which were then induced for the production of the recombinant HN and F proteins. However, there was no protein expression observed in the recombinant P. pastoris and S. cerevisiae cells. Whereas, the bacterial hosts were found expressing the recombinant HN and F proteins (from the pRSETA/HN and pRSETB/F plasmids respectively), and the NusA fusion proteins, NusA-HN and NusA-F (from the pET-43.1a/HN and pET-43.1a/F plasmids respectively). The recombinant HN and F proteins were produced as insoluble inclusion bodies (IB) while the NusA-HN and NusA-F proteins were expressed in soluble form in E. coli. The recombinant proteins were purified and used to immunise specific pathogen-free (SPF) chickens. ELISA results revealed that the insoluble and urea-solubilised inclusion bodies of the recombinant HN and F proteins, and the soluble NusA-HN and NusA-F proteins stimulated the production of antibodies that detect NDV. Among these antigens, the urea-solubilised HN IB appeared to induce the highest antibody titers. However, the chicken antibodies failed to neutralise the viral activities as shown in the tests such as haemagglutination inhibition (HI), neuraminidase inhibition (NI) and haemolysis inhibition (HLI). This explains the susceptibility of the immunised flocks to NDV infection upon the viral challenge. Despite of the presence of antibodies to NDV, none of the immunised chicken was protected against the viral challenge. Immunoblotting analysis on the interactions between the antigens and antibodies revealed that the anti-F antibodies did not bind to the denatured viral F glycoprotein, neither the anti-NDV serum detect the recombinant F protein. However, the anti-HN antibodies showed positive signals when used to probe the denatured viral HN glycoprotein, and in return,the anti-NDV serum detected the recombinant HN protein. This finding indicates the potential application of the E. coli produced HN protein as antigen for the detection of NDV antibody

Diversity of edible mollusc (Gastropoda and Bivalve) at selected division of Sarawak, Malaysia.

Diversity of edible mollusc was studied at eight divisions of Sarawak from August 2010 to May 2011. At each division, diversity and number of species were collected from road site selling out lets and local wet markets. Total number of mollusc was comprised of 29 species namely Solen regularies, S. lamarckii, Pharella acutidens, Anadara granosa, Pholas orientalis, Gluconome virens, Circe scripta, Anodonta woodina, Paphia undulata, Amusium pleuronectes, Meretrix meretrix, M. lyrata, Polymesoda bengalensis, P. erosa and P. expansa for bivalve and Cerithidea rizophorarum, C. obtusa, Telescopium telescopium, Clithon retropictus, Nerita articulate, N. chamaeleon, N. albicilla, Ellobium aurisjuda, Trochus radiates, Planaxis sulcatus, Monodonta labio, Turbo crasus, Thais aculate and Melo melo for gastropod. The species number of bivalve was recorded highest (15 species) in mollusc group from Sarawak. Mollusc diversity was found highest (14 species) in Bintulu and lowest (1 species) in the division of Sarikei. From this study, there is a wide chance of research to further explore both on the possibility of commercial value and ecosystem conservation

Taxonomic study of edible bivalve from selected division of Sarawak, Malaysia.

The diversity of edible bivalve was conducted from August 2010 to July 2011 covering eight divisions i.e., Kuching, Sarikei, Sibu, Mukah, Bintulu, Miri, Limbang and Lawas of Sarawak, Malaysia. Samples were collected from native market and fishing village during the study period. All edible bivalves inhabit either in brackish or marine environment and comprised 19 species from 10 families namely Meretrix meretrix, M. lyrata, Paphia undulata, Circe scripta, Solen regularies, Solen lamarckii, Pharella acutidens, Amusium pleuronectes, Anadara granosa, Pholas orientalis, Gluconome virens, Placuna placenta, Crassotrea lugubris, Isognomon ephippium, Polymesoda erosa, P. bengalensis, P. expansa, Anadonta woodina and Pilsbryoconcha exilis. The diversity of edible bivalves was found highest in Kuching and Bintulu compared to other divisions studied in Sarawak. The bivalve species at Sarawak could have economic potentiality in terms of protein source, livelihoods of local tribes and economic value. Study suggests that if the high conservation and management of edible bivalve diversity could establish in the coastal and wetland area of Sarawak, a remarkable and vast economic return could achieve

Diversity of Edible Mollusc (Gastropoda and Bivalvia) at Selected Divison of Sarawak, Malaysia

Diversity of edible mollusc was studied at eight divisions of Sarawak from August 2010 to May 2011. At each division, diversity and number of species were collected from road site selling out lets and local wet markets. Total number of mollusc was comprised of 29 species namely  Solen regularies, S. lamarckii,  Pharella acutidens, Anadara granosa, Pholas orientalis, Gluconome virens, Circe scripta, Anodonta woodina, Paphia undulata, Amusium pleuronectes, Meretrix meretrix, M. lyrata, Polymesoda bengalensis, P. erosa and P. expansa  for bivalve and  Cerithidea rizophorarum, C. obtusa, Telescopium telescopium, Clithon retropictus, Nerita articulate, N. chamaeleon, N. albicilla, Ellobium aurisjuda, Trochus radiates, Planaxis sulcatus, Monodonta labio, Turbo crasus, Thais aculate  and Melo melo  for gastropod. The species number of bivalve was recorded highest (15 species) in mollusc group from Sarawak. Mollusc diversity was found highest (14 species) in Bintulu and lowest (1 species) in the division of Sarikei. From this study, there is a wide chance of research to further explore both on the possibility of commercial value and ecosystem conservation

Diversity of cephalopod from selected division of Sarawak, Malaysia.

The diversity of cephalopod from nine divisions at Sarawak, Malaysia was studied from August 2010 to February 2011. Cephalopod was collected from landing stations and local wet markets of Kuching, Sri Aman, Sibu, Sarikei, Mukah, Miri, Limbang, Lawas and Bintulu. Seven species of cephalopod belongings to three families were identified during the study period. The family Loliginidae was dominated group of cephalopod which consists of three species namely Uroteuthis (Photololigo) edulis, Uroteuthis (Photololigo) chinensis and Sepioteuthis lessoniana and recorded from five divisions. The species of cuttlefishes (Family Sepiidae) were Sepia recurvirostra, Sepia aculeate and Sepiella inermis and recorded from Sarikei, Kuching and Bintulu. Octopodidae was the only one species named Cistopus indicus and found in Bintulu. The diversity of cephalopod was found highest in Bintulu with seven species while surprisingly no species of cephalopod was recorded from Limbang, Lawas and Mukah. Study suggests that the findings of this study could be useful for future reference and detail study on cephalopod in the coastal water of Sarawak, Malaysia

Taxonomic assessment of seaweed community from the coastal areas of Bintulu, Sarawak, Malaysia

Basic taxonomic information forms the important basis for the documentation, resource management and utilization of marine biodiversity such as seaweeds. A taxonomic assessment of seaweeds in the coastal areas of Bintulu, Sarawak, East Malaysia, was conducted monthly from May 2011 to May 2012. Species composition was recorded following NaGISA protocols, direct observation, and SCUBA and snorkeling techniques. A total of 54 species were identified, classified into Rhodophyta (23 species), Chlorophyta (16 species) and Phaeophyta (15 species). The highest abundance was recorded at Kuala Similajau (25 species) while the lowest was recorded at Kuala Nyalau (12 species). As the present study was conducted by examining species collected from both rocky shores and the reef area for the first time, a higher number of species was documented compared to previous studies conducted in the same general area but focusing only on particular habitats.Thirty species found in the current survey represent new records for the locality including some with economic potentials

Diversity of Cephalopod from Selected Division of Sarawak, Malaysia

The diversity of cephalopod from nine divisions at Sarawak, Malaysia was studied from August 2010 to February 2011. Cephalopod was collected from landing stations and local wet markets of Kuching, Sri Aman, Sibu, Sarikei, Mukah, Miri, Limbang, Lawas and Bintulu. Seven species of cephalopod belongings to three families were identified during the study period. The family Loliginidae was dominated group of cephalopod which consists of three species namely  Uroteuthis (Photololigo) edulis, Uroteuthis (Photololigo) chinensis and Sepioteuthis lessoniana and recorded from five divisions. The species of cuttlefishes (Family Sepiidae) were Sepia recurvirostra, Sepia aculeate and Sepiella inermis and recorded from Sarikei, Kuching and Bintulu. Octopodidae was the only one species named Cistopus indicus and found in Bintulu. The diversity of cephalopod was found highest in Bintulu with seven species while surprisingly no species of cephalopod was recorded from Limbang, Lawas and Mukah. Study suggests that the findings of this study could be useful for future reference and detail study on cephalopod in the coastal water of Sarawak, Malaysia

Terrestrial green algae diversity in oil palm plantation and secondary forest using 18S-rDNA PCR-DGGE approach

The demand for vegetables oils especially palm oil has increased drastically. However, rapid development in oil palm industry leads to many issues, especially regarding sustainable development of palm oil cultivation. Soil algae play an important role in the maintenance of soil fertility as well as recycling of soil organic matter. Their taxonomic knowledge in tropical soils is still scarce, especially in oil palm plantation. Hence, this study was performed to access the heterogeneity of soil algae community structure in response to oil palm plantation activities and compare with nearby secondary forest by using 18S-rDNA PCR-DGGE. Sampling was carried out in 3-years old oil palm plantation at Sungai Asap, Sarawak and nearby disturbed and undisturbed secondary forest. Significant differences in soil chemical properties and soil texture of oil palm plantation were evident: soil organic C, total N and CEC were significantly decreased. Oil palm plantation showed highest richness and diversity of the green algae, followed by disturbed secondary forest and undisturbed secondary forest. Three main clusters were obtained through UPGMA dendrogram generated with dice coefficient, one cluster from each sampling sites, except for some oil palm plantation sampling points were in the same clusteras with the disturbed secondary forest. Collectively, these results revealed that disturbance by both human or agricultural activity altered soil algae richness and diversity

Effect of fermented kitchen waste on tilapia (Oreochromis niloticus) growth performance and water quality as a water additive

Aquaculture contributes about 20% of domestic fish production in Malaysia. Tilapia has been identified as one of the main species for freshwater aquaculture in the Third National Agriculture Policy (DPN3). However, feed cost and water quality management remain as two major challenges to the industry. This study aim to analyse the effects of Fermented Kitchen Waste (FKW) as water additives on water quality and growth performance of tilapia (Oreochromis niloticus). Different concentration (0.05, 0.1 and 0.2%) of FKW were used to treat tilapia in tank culture for a period of twelve weeks. Physico-chemical parameters were also taken every week. Treatment with 0.1% FKW resulted in significant (p<0.05) decrease in ammonia, nitrite and nitrate levels. The survival rates of tilapia treated with 0.05 and 0.1% FKW were comparable to the untreated control. Growth performance of the tilapia was measured in term of length and weight. Highest relative growth rate was observed in tilapia treated with 0.05% FKW. However, all the fish died in 0.2% FKW due to severe pH drop. Therefore, low concentration of FKW could severe as a potential water additive to improve water quality and promote growth in tilapia aquaculture