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.     

Extracellular hydrolase enzyme production by soil fungi from King George Island, Antarctica

Various microbial groups are well known to produce a range of extracellular enzymes and other secondary metabolites. However, the occurrence and importance of investment in such activities have received relatively limited attention in studies of Antarctic soil microbiota. In order to examine extracellular enzyme production in this chronically low-temperature environment, fungi were isolated from ornithogenic, pristine and human-impacted soils collected from the Fildes Peninsula, King George Island, Antarctica during the austral summer in February 2007. Twenty-eight isolates of psychrophilic and psychrotolerant fungi were obtained and screened at a culture temperature of 4°C for activity of extracellular hydrolase enzymes (amylase, cellulase, protease), using R2A agar plates supplemented with (a) starch for amylase activity, (b) carboxymethyl cellulose and trypan blue for cellulase activity or (c) skim milk for protease activity. Sixteen isolates showed activity for amylase, 23 for cellulase and 21 for protease. One isolate showed significant activity across all three enzyme types, and a further 10 isolates showed significant activity for at least two of the enzymes. No clear associations were apparent between the fungal taxa isolated and the type of source soil, or in the balance of production of different extracellular enzymes between the different soil habitats sampled. Investment in extracellular enzyme production is clearly an important element of the survival strategy of these fungi in maritime Antarctic soils.     

Plasmodium knowlesi: reservoir hosts and tracking the emergence in humans and macaques

Plasmodium knowlesi, a malaria parasite originally thought to be restricted to macaques in Southeast Asia, has recently been recognized as a significant cause of human malaria. Unlike the benign and morphologically similar P. malariae, these parasites can lead to fatal infections. Malaria parasites, including P. knowlesi, have not yet been detected in macaques of the Kapit Division of Malaysian Borneo, where the majority of human knowlesi malaria cases have been reported. In order to extend our understanding of the epidemiology and evolutionary history of P. knowlesi, we examined 108 wild macaques for malaria parasites and sequenced the circumsporozoite protein (csp) gene and mitochondrial (mt) DNA of P. knowlesi isolates derived from macaques and humans. We detected five species of Plasmodium (P. knowlesi, P. inui, P. cynomolgi, P. fieldi and P. coatneyi) in the long-tailed and pig-tailed macaques, and an extremely high prevalence of P. inui and P. knowlesi. Macaques had a higher number of P. knowlesi genotypes per infection than humans, and some diverse alleles of the P. knowlesi csp gene and certain mtDNA haplotypes were shared between both hosts. Analyses of DNA sequence data indicate that there are no mtDNA lineages associated exclusively with either host. Furthermore, our analyses of the mtDNA data reveal that P. knowlesi is derived from an ancestral parasite population that existed prior to human settlement in Southeast Asia, and underwent significant population expansion approximately 30,000-40,000 years ago. Our results indicate that human infections with P. knowlesi are not newly emergent in Southeast Asia and that knowlesi malaria is primarily a zoonosis with wild macaques as the reservoir hosts. However, ongoing ecological changes resulting from deforestation, with an associated increase in the human population, could enable this pathogenic species of Plasmodium to switch to humans as the preferred host.     

Synergism between curdlan and GM-CSF confers a strong inflammatory signature to dendritic cells

A simultaneous engagement of different pathogen recognition receptors provides a tailor-made adaptive immunity for an efficient defense against distinct pathogens. For example, cross-talk of TLR and C-type lectin signaling effectively shapes distinct gene expression patterns by integrating the signals at the level of NF-κB. In this study, we extend this principle to a strong synergism between the dectin-1 agonist curdlan and an inflammatory growth factor, GM-CSF. Both together act in synergy in inducing a strong inflammatory signature that converts immature dendritic cells (DCs) to potent effector DCs. A variety of cytokines (IL-1β, IL-6, TNF-α, IL-2, and IL-12p70), costimulatory molecules (CD80, CD86, CD40, and CD70), chemokines (CXCL1, CXCL2, CXCL3, CCL12, CCL17), as well as receptors and molecules involved in fugal recognition and immunity such as Mincle, dectin-1, dectin-2, and pentraxin 3 are strongly upregulated in DC treated simultaneously with curdlan and GM-CSF. The synergistic effect of both stimuli resulted in strong IκBα phosphorylation, its rapid degradation, and enhanced nuclear translocation of all NF-κB subunits. We further identified MAPK ERK as one possible integration site of both signals, because its phosphorylation was clearly augmented when curdlan was coapplied with GM-CSF. Our data demonstrate that the immunomodulatory activity of curdlan requires an additional signal provided by GM-CSF to successfully initiate a robust β-glucan-specific cytokine and chemokine response. The integration of both signals clearly prime and tailor a more effective innate and adaptive response against invading microbes and fungi.     

Geraniin extracted from the rind of <Emphasis Type="Italic">Nephelium lappaceum</Emphasis> binds to dengue virus type-2 envelope protein and inhibits early stage of virus replication

Background

The rapid rise and spread in dengue cases, together with the unavailability of safe vaccines and effective antiviral drugs, warrant the need to discover and develop novel anti-dengue treatments. In this study the antiviral activity of geraniin, extracted from the rind of Nephelium lappaceum, against dengue virus type-2 (DENV-2) was investigated.

Methods

Geraniin was prepared from Nephelium lappaceum rind by reverse phase C-18 column chromatography. Cytotoxicity of geraniin towards Vero cells was evaluated using MTT assay while IC₅₀ value was determined by plaque reduction assay. The mode-of-action of geraniin was characterized using the virucidal, attachment, penetration and the time-of-addition assays’. Docking experiments with geraniin molecule and the DENV envelope (E) protein was also performed. Finally, recombinant E Domain III (rE-DIII) protein was produced to physiologically test the binding of geraniin to DENV-2 E-DIII protein, through ELISA competitive binding assay.

Results

Cytotoxicity assay confirmed that geraniin was not toxic to Vero cells, even at the highest concentration tested. The compound exhibited DENV-2 plaque formation inhibition, with an IC₅₀ of 1.75 μM. We further revealed that geraniin reduced viral infectivity and inhibited DENV-2 from attaching to the cells but had little effect on its penetration. Geraniin was observed to be most effective when added at the early stage of DENV-2 infection. Docking experiments showed that geraniin binds to DENV E protein, specifically at the DIII region, while the ELISA competitive binding assay confirmed geraniin’s interaction with rE-DIII with high affinity.

Conclusions

Geraniin from the rind of Nephelium lappaceum has antiviral activity against DENV-2. It is postulated that the compound inhibits viral attachment by binding to the E-DIII protein and interferes with the initial cell-virus interaction. Our results demonstrate that geraniin has the potential to be developed into an effective antiviral treatment, particularly for early phase dengue viral infection.

     

Complex combination of seed dormancy and seedling development determine emergence of Viburnum tinus (Caprifoliaceae)

BACKGROUND AND AIMS: The shrub Viburnum tinus is widely distributed in mattoral vegetation of the Mediterranean basin. The purpose of the present study was to classify the seed dormancy type and examine the requirements for embryo growth, root protrusion and shoot emergence. METHODS: Overwintered fruits were collected in western Spain in April 2001 and prepared in three ways: entire pericarp was removed, exocarp and mesocarp were removed or fruits were left intact. Fruits treated in these three ways were subjected to artificial annual temperature cycles or to constant temperature regimes for 1.5 years. KEY RESULTS: Removal of exocarp and mesocarp was necessary for embryo growth and germination. High temperature favoured dormancy alleviation and embryo growth, intermediate to low temperatures favoured root protrusion, and intermediate temperature shoot emergence. There was substantial germination at constant temperature regimes, indicating an overlap between temperature intervals suitable for the different stages of embryo and seedling development. Functionally, V. tinus has the same root and shoot emergence pattern that is described for other Viburnum species considered to have epicotyl dormancy. However, the requirement for high and low temperatures for radicle protrusion and epicotyl emergence, respectively, was missing in V. tinus; these characters are the foundation for the epicotyl dormancy classification. CONCLUSIONS: It is concluded that V. tinus does not have epicotyl dormancy. Instead, there is a combination of a weak morphophysiological dormancy and a slow germination process, where different temperatures during an annual cycle favour different development stages. The present study suggests that the first complete seedlings would emerge in the field 1.5 years after fruit maturation in October, i.e. seed dispersal during winter, embryo growth during the first summer, root protrusion and establishment during the second autumn and winter, and cotyledon emergence during the second spring.     

Correlation of SOD and MDA Expression in the Organ of Corti and Changes in the Function of Outer Hair Cells Measured by DPOAE Examination in Noise-Exposed Rat Cochlea

Background:Hearing loss due to noise can cause the disturbances toward the quality of life and cause mechanical damage and metabolic decompensation. Distortion Product Otoacoustic Emission (DPOAE) is an examination to assess the sensory function of outer hair cells. Superoxide Dismutase (SOD) and Malondialdehyde (MDA) are markers of oxidative stress. The aim of this study was to identify the correlation between DPOAE examination and SOD and MDA expression in rats exposed to noise.Methods:This research was conducted on 27 rats which were divided into 3 groups, group 1 (control), group 2, and group 3 were groups with 100 dB and 110 dB noise exposure respectively.Results:Our findings show a decrease in SOD expression and DPOAE values as well as an increase in MDA expression in rats exposed to noise and there is a positive correlation between Signal to Noise Ratio (SNR) value with SOD expression (r= 0.733, p= 0.025) and a negative correlation between SNR value with MDA expression (r= -0.678, p= 0.045).Conclusion:our study find the correlation of oxidant and antioxidant status values in the organ of corti and changes in the function of outer hair cells in noise-exposed rat models.Key Words: Malondialdehyde, Noise Induced Hearing Loss, Otoacoustic Emission, Superoxide Dismutase     

Insecticide resistance in dengue vectors from hotspots in Selangor,Malaysia

BackgroundIn Malaysia, dengue remains a top priority disease and usage of insecticides is the main method for dengue vector control. Limited baseline insecticide resistance data in dengue hotspots has prompted us to conduct this study. The present study reports the use of a map on the insecticide susceptibility status of Aedes aegypti and Aedes albopictus to provide a quick visualization and overview of the distribution of insecticide resistance.Method and resultsThe insecticide resistance status of Aedes populations collected from 24 dengue hotspot areas from the period of December 2018 until June 2019 was proactively monitored using the World Health Organization standard protocol for adult and larval susceptibility testing was conducted, together with elucidation of the mechanisms involved in observed resistance. For resistance monitoring, susceptibility to three adulticides (permethrin, deltamethrin, and malathion) was tested, as well as susceptibility to the larvicide, temephos. Data showed significant resistance to both deltamethrin and permethrin (pyrethroid insecticides), and to malathion (organophosphate insecticide) in all sampled Aedes aegypti populations, while variable resistance patterns were found in the sampled Aedes albopictus populations. Temephos resistance was observed when larvae were tested using the diagnostic dosage of 0.012mg/L but not at the operational dosage of 1mg/L for both species.ConclusionThe present study highlights evidence of a potential threat to the effectiveness of insecticides currently used in dengue vector control, and the urgent requirement for insecticide resistance management to be integrated into the National Dengue Control Program.     

Genetic variation in Malaysian oysters: taxonomic ambiguities and evidence of biological invasion

Genetic diversities in two cultured oyster species, Crassostrea iredalei (Faustino 1932) and Crassostrea belcheri (Sowerby 1871) were assessed using a 581-nucleotide fragment of the mtDNA cytochrome oxidase subunit 1 (CO1) gene. A total of 103 C. iredalei individuals and 120 C. belcheri from 12 populations were sampled along the coast of Malaysia. Trees of unique haplotype samples generated based on Neighbor-Joining (NJ) algorithm revealed that many individuals had been misidentified and did not cluster with their presumed species based on morphological identification. BLAST results of DNA sequences showed presence of previously unreported C. madrasensis in Peninsular Malaysian waters (98% maximum identity). The true identity of the Muar (Crassostrea sp.) and Semporna (Saccostrea sp.) populations were unresolved by two BLAST search and showed less than 88% identity with other species in GenBank. Repeated analysis of these two populations using 487 bp of the mitochondrial 16S gene data showed only a maximum identity less than 97%. Hence, the identity of these specimens remains unclear. Evolutionary divergences within presumed species were 0.001–0.011 and 0.034–0.313 between species. Findings from this study have important implications for aquaculture, management and monitoring of cultured populations as well as conservation of wild oyster species in Malaysia.     

Morphometric and molecular analysis of mackerel (Rastrelliger spp) from the west coast of Peninsular Malaysia

Mackerel (Scombridae; Rastrelliger) are small commercially important pelagic fish found in tropical regions. They serve as a cheap source of animal protein and are commonly used as live bait. By using a truss morphometrics protocol and RAPD analysis, we examined morphological and genetic variation among 77 individual mackerel that were caught using long lines and gillnets at 11 locations along the west coast of Peninsular Malaysia. Nineteen morphometric traits were evaluated and genetic information was estimated using five 10-base RAPD random primers. Total DNA was extracted from muscle tissue. Morphometric discriminant function analysis revealed that two morphologically distinct groups of Rastrelliger kanagurta and a single group of R. brachysoma can be found along the west coast of Peninsular Malaysia. We also found that the head-related characters and those from the anterior part of the body of Rastrelliger spp significantly contribute to stock assessment of this population. RAPD analysis showed a trend similar to that of the morphometric analysis, suggesting a genetic component to the observed phenotypic differentiation. These data will be useful for developing conservation strategies for these species.