Generation by Reverse Genetics of an Effective,Stable, Live-Attenuated Newcastle Disease Virus Vaccine Based on a Currently Circulating,Highly Virulent Indonesian Strain

Newcastle disease virus (NDV) can cause severe disease in chickens. Although NDV vaccines exist, there are frequent reports of outbreaks in vaccinated chickens. During 2009–2010, despite intense vaccination, NDV caused major outbreaks among commercial poultry farms in Indonesia. These outbreaks raised concern regarding the protective immunity of current vaccines against circulating virulent strains in Indonesia. In this study, we investigated whether a recombinant attenuated Indonesian NDV strain could provide better protection against prevalent Indonesian viruses. A reverse genetics system for the highly virulent NDV strain Banjarmasin/010/10 (Ban/010) isolated in Indonesia in 2010 was constructed. The Ban/010 virus is classified in genotype VII of class II NDV, which is genetically distinct from the commercial vaccine strains B1 and LaSota, which belong to genotype II, and shares only 89 and 87% amino acid identity for the protective antigens F and HN, respectively. A mutant virus, named Ban/AF, was developed in which the virulent F protein cleavage site motif “RRQKR↓F” was modified to an avirulent motif “GRQGR↓L” by three amino acid substitutions (underlined). The Ban/AF vaccine virus did not produce syncytia or plaques in cell culture, even in the presence of added protease. Pathogenicity tests showed that Ban/AF was completely avirulent. Ban/AF replicated efficiently during 10 consecutive passages in chickens and remained genetically stable. Serological analysis showed that Ban/AF induced higher neutralization and hemagglutination inhibition antibody titers against the prevalent viruses than the commercial vaccines B1 or LaSota. Both Ban/AF and commercial vaccines provided protection against clinical disease and mortality after challenge with virulent NDV strain Ban/010 (genotype VII) or GB Texas (genotype II). However, Ban/AF significantly reduced challenge virus shedding from the vaccinated birds compared to B1 vaccine. These results suggest that Ban/AF can provide better protection than commercial vaccines and is a promising vaccine candidate against NDV strains circulating in Indonesia.     

The abundance and diversity of Odonata along an altitudinal gradient in East Java,Indonesia

Odonate diversity in East Java was surveyed, and samplings were made during 2 years in ten sites along an altitudinal gradient. The characteristics of odonate assemblages in East Java were analyzed regarding of the number of individuals, the number of species, and Shannon's diversity index. The differences in abundance, species richness, and diversity between study sites were analyzed by an independent t‐test. There were 3270 individuals of Odonata belong to 30 species, 7 families and 2 suborders identified from all study sites. The abundance, species richness and diversity of Odonata varied between study sites. The greatest abundance of Odonata was found in Malang Coban Talun (MCT) (148.8 ± 9.5), while the lowest was in South Beach Forest (SBF) (14.4 ± 3.6). The highest species richness and diversity was found in Malang Paddy Field (MPF) (richness =14.4 ± 0.8 and H′ = 2.4 ± 0.1), while the lowest was found in South Coastal Area (SCA) (richness =2.8 ± 0.1 and H′ = 0.4 ± 0.1). A significant positive correlation was detected between the elevation and overall odonate abundance (P < 0.05), while there was not significant correlation between that and odonate species richness and diversity.     

Loss of Protein Kinase C-δ Protects against LPS-Induced Osteolysis Owing to an Intrinsic Defect in Osteoclastic Bone Resorption

Bone remodeling is intrinsically regulated by cell signaling molecules. The Protein Kinase C (PKC) family of serine/threonine kinases is involved in multiple signaling pathways including cell proliferation, differentiation, apoptosis and osteoclast biology. However, the precise involvement of individual PKC isoforms in the regulation of osteoclast formation and bone homeostasis remains unclear. Here, we identify PKC-δ as the major PKC isoform expressed among all PKCs in osteoclasts; including classical PKCs (−α, −β and −γ), novel PKCs (−δ, −ε, −η and −θ) and atypical PKCs (−ι/λ and −ζ). Interestingly, pharmacological inhibition and genetic ablation of PKC-δ impairs osteoclastic bone resorption in vitro. Moreover, disruption of PKC-δ activity protects against LPS-induced osteolysis in mice, with osteoclasts accumulating on the bone surface failing to resorb bone. Treatment with the PKC-δ inhibitor Rottlerin, blocks LPS-induced bone resorption in mice. Consistently, PKC-δ deficient mice exhibit increased trabeculae bone containing residual cartilage matrix, indicative of an osteoclast-rich osteopetrosis phenotype. Cultured ex vivo osteoclasts derived from PKC-δ null mice exhibit decreased CTX-1 levels and MARKS phosphorylation, with enhanced formation rates. This is accompanied by elevated gene expression levels of cathepsin K and PKC −α, −γ and −ε, as well as altered signaling of pERK and pcSrc416/527 upon RANKL-induction, possibly to compensate for the defects in bone resorption. Collectively, our data indicate that PKC-δ is an intrinsic regulator of osteoclast formation and bone resorption and thus is a potential therapeutic target for pathological osteolysis.     

Using green fluorescent protein sheds light on <Emphasis Type="Italic">Lupinus angustifolius</Emphasis> L. transgenic shoot development

Narrow-leaf lupin (NLL) is the main legume crop grown in rotation with wheat and other cereals in Western Australia. Efforts to improve NLL germplasm by use of genetic technologies have been hampered by the lack of an efficient genetic transformation method, an issue that is in common with dominant crop legumes globally. Prior research has primarily used the bar gene for phosphinothricin (PPT) resistance. The aim of recent research has been to investigate alternative selection methodologies, in order to determine whether the limitations of low frequency of transgenic shoots, combined with chimerism at T0 could be overcome. Investigation of hygromycin resistance as a selectable marker compared to PPT is reported here. The results suggested that hygromycin resistance was a more suitable selectable marker for NLL transformation than PPT. Surprisingly, from investigation of transformation using the GUS reporter gene, it was also observed that transformation frequency was greater when selection treatment was reduced or delayed, compared to the existing protocols. To further investigate this observation, an eGFP expressing construct was prepared. Observations within the first week after Agrobacterium exposure of lupin explants demonstrated that transformation of NLL explant cells was not a rate-limiting step. Instead, the results indicated that the current selection methodology was killing the cells that were competent to regenerate into transgenic shoots. It was concluded that further research on the development of the treated explants should focus on delayed selection and exposure to Agrobacterium of cells below the apical meristem.     

Food sources and trophic structure of fishes and benthic macroinvertebrates in a tropical seagrass meadow revealed by stable isotope analysis

Stable carbon and nitrogen isotope analysis was used to examine the food sources and trophic structure of 17 fish species and six groups of benthic macroinvertebrates in a seagrass meadow in North Sulawesi, Indonesia. The seagrass, their associated epiphytes, sediment organic matter (SOM) and particulate organic matter (POM) were identified to be the food sources, with δ¹³C values ranging from ?19.49 (POM) to ?9.66‰ (seagrass). The δ¹³C of the 23 fauna taxa were between ?18.57 (Arothron manilensis) and ?11.62‰ (Protoreaster sp.). For five of the six groups of benthic macroinvertebrates, seagrass and their epiphytes contributed more than 69.4%. For 14 of the 17 fish species, seagrass and their epiphytes are the main contributors. For 15 of the 17 fishes, the trophic levels inferred from SIA are lower than those from the previously reported diet composition analysis. These findings show that seagrass and their epiphytes are consumed by most of the fish and benthic macroinvertebrates, and are important for a large portion of the food web in seagrass meadows in the Coral Triangle area.     

The <Emphasis Type="Italic">Leiognathus aureus</Emphasis> complex (Perciformes: Leiognathidae) with the description of a new species

Taxonomic analysis of a group of morphologically similar ponyfishes (Perciformes: Leiognathidae) establishes a complex comprising three valid species: Leiognathus aureus Abe and Haneda, 1972, widely distributed in the western Pacific Ocean (Taiwan, Philippines, Thailand, Singapore, Indonesia, and northern Australia); L. hataii Abe and Haneda, 1972, currently known only from Ambon, Indonesia; and L. panayensis sp. nov. Kimura and Dunlap, currently known only from Panay Island, the Philippines. The L. aureus complex can be defined by the following combination of characters: mouth protruding forward, not downward; small but sharp conical teeth uniserially on jaws; a black line between lower margin of eye and lower jaw articulation; and lateral line incomplete, ending below posterior part of dorsal fin base or on anterior caudal peduncle. Leiognathus hataii differs from both L. aureus and L. panayensis in having a large dark blotch below the spinous dorsal fin base and fewer counts of scales (lateral line scales 50–58 vs. 64–85 in the latter two species; scales above lateral line 7–10 vs. 12–18; scales below lateral line 22–26 vs. 30–41). Leiognathus panayensis is distinguished from L. aureus in having a deeper body (41–51% SL vs. 35–45% SL in the latter), long posterior limb of maxilla (21–25% HL vs. 15–23% HL), wholly scaled belly (vs. naked along preanal median keel), and a dark blotch on nape (vs. absent).