Seasonal variation of fatty acid content in natural microplankton from the Tumpat coastal waters of the South China Sea

In the search for better understanding on the nutritional quality of natural tropical plankton, samples were collected from shallow coastal waters facing the South China Sea during the dry monsoon (May-September) and the wet monsoon (November-April) seasons from March 1993 to July 1994. The total fatty acid content of the predominantly phytoplankton communities (25-200 microns sieve nets) varied four to fivefold with the lowest value occurring during the dry monsoon when blue-green became predominant. Saturated fatty acid content (SAFA), polyunsaturated fatty acid (PUFA) and total omega 3 (sigma omega 3) showed the same seasonal pattern as the total fatty acid with high values in October to December 1993. When species of the dinoflagellate Peridinium and Ceratium were present in considerable amount, the docosahexaenoic acid DHA content was high, especially from March to May 1993. The maximum content of eicosapentaenoic acid EPA, total omega-3 fatty acid, PUFA and sigma omega 3 in phytoplankton occurred during the pre-monsoon period (October and November 1993) when the diatoms were present in large amounts. The larger fraction sample (> 200 microns sieve nets) which consisted predominantly of zooplankton had high amounts of PUFA from September to November 1993.     

Seasonal changes in composition and biomass of epiphytic algae on the macrophyte Ranunculus penicillatus in a chalk stream,with estimates of production,and observations on the epiphytes of Cladophora glomerata

The epiphytic algae on surfaces of the macrophyte Ranunculus penicillatus (Dumort.) Bab. var. calcareus (R. W. Butcher) C. D. K. Cook and on Cladophora glomerata (L.) Kütz growing in the River Itchen at Otterbourne near Southampton were studied between February 1984 and June 1985. The river at this site has a mean flow rate of 0.33 m s^–1, and is about 16 m wide and on average 20 cm deep, with a discharge ranging through the year between 0.34 and 2.46 m³ s^–1. The pH of the river varies little around 8.2, with a mean alkalinity of 236 mg HCO _inf3 ^sup1 l^–1, because of its origin from chalk springs. Ranunculus grows throughout the year, with peaks of biomass in spring and autumn. It forms a very large surface for attachment of epiphytes, and covers on average 40% of the stream bed. Numbers of epiphyte cells removed from Ranunculus ranged through the year between 52 × 10³ and 271 × 10³ cells mm^–2 stream floor, with maximum numbers in April, and a secondary peak in October. This pattern partly reflects fluctuations in the biomass of Ranunculus; the number of cells per unit area of plant surface showed a broader spring peak and lower fluctuations in other seasons. Diatoms formed 65 to 98% of these epiphyte cells, with chlorophytes reaching their peak (10%) in summer and cyanophytes (25%) in autumn. Estimates of biomass of these epiphytic forms, derived from measurements of chlorophyll c, indicate a range between 30 and 100 g dry weight m^–2 of weed bed. Colonisation studies showed that the algae grow and reproduce throughout the year, with a mean generation time of about 5 days, suggesting an annual production of about 3 kg dry weight m^–2 of weed bed, which makes epiphytic algae the principal primary producers in the stream. The numbers and biomass of epiphytic algae on Cladophora are considerably less.The species of epiphytic algae found on Ranunculus were generally similar to those growing on Cladophora, and to epilithic algae on pebbles of the stream bed, but different species were dominant on the different substrata. Algal cells in the water column were all derived from benthic habitats, although their relative abundance was very different.     

Seasonal changes in composition and biomass of epilithic algal floras of a chalk stream and a soft water stream with estimates of production

Microscopic epilithic algae in the River Itchen at Otterbourne near Southampton and in the Ober Water in the New Forest were studied during 1984 and 1985. The River Itchen rises from chalk springs and has a steady pH near 8.2 and a mean alkalinity of 236 mg HCO₃ 1^–1; at the study site the river is about 16 m wide and 20 cm deep, with a mean flow rate of 0.33 m s^–1 and a discharge ranging through the year between 0.34 and 2.46 m³ s^–1. The Ober Water, which drains sands and gravels, has a pH between 6.9 and 7.2 and a mean alkalinity of about 50 mg HCO₃ 1^–1; at the study site it is about 6 m wide, with a mean flow rate of 0.27 m s^–1 and a discharge ranging through the year between 0.08 and 1.0 m³ s^–1.Epilithic algae removed from the pebbles that form the major part of the beds of both streams show seasonal changes in abundance and composition. Diatoms peaked in April/May and dominate the epilithic flora in both streams, comprising 70–95% of all algal cells; highest numbers of chlorophytes occurred in summer and cyanophytes increased in autumn. The species composition of the epilithic flora in the two streams was different, as was the population density; algal cell numbers ranged between 500 and 7000 cells mm^–2 of stream floor in the River Itchen and between 8 and 320 cells mm^–2 of stream floor in the Ober Water. The chlorophyll a content of epilithic algae in the River Itchen ranged between 115 and 415 mg m^–2 of stream floor, representing an annual mean biomass of about 8 g m^–2, whereas in the Ober Water a chlorophyll a content of 2.2 to 44 mg m^–2 of stream floor was found, representing an annual mean biomass of about 1 g m^–2. Cautious estimates of the annual production of epilithic algae in these streams suggest a value of about 600 g organic dry weight m^–2 in the River Itchen and about 75 g m^–2 in the Ober Water.     

In silico identification and characterization of a putative phosphatidylinositol 4-phosphate 5-kinase (PIP5K) gene in Eimeria tenella

Phosphatidylinositol 4-phosphate 5-kinases (PIP5Ks) play diverse roles in the cellular biology of many organisms, including signal transduction, secretion and vesicular trafficking, and regulation of cytoskeleton assembly. Discovery of the PIP5K gene in Eimeria tenella may shed light on its role in the biology of this avian protozoan, and afford further understanding of the cell-host interaction, particularly during the invasion process. In this study, we report the identification of the PIP5K coding region in the genome sequence of Eimeria tenella using in silico gene prediction approaches. Prediction of the PIP5K coding sequence was confirmed by mapping the full-length cDNA sequence, generated via the Rapid Amplification of cDNA Ends (RACE) method, to the genomic sequence. The putative PIP5K gene of Eimeria tenella is located on the complementary strand of the E1080B12.b1 contig, and comprises 12 exons. Further analysis showed that the coding region spans from exon 1 to exon 7, with all exons obeying the adopted 'gt...ag' splicing rule of intronic sequences. Consensus of the hexameric 5' donor-splice site was deduced as GTRDBB... and the consensus for the 3' acceptor-splice sites as ...BHDYAG. The gene encodes a 252-amino acid residue protein. Domain search and protein fold recognition analyses provide compelling evidences that the deduced protein is a PIP5K.     

Lipid and fatty acid composition of microalgae used in Malaysian aquaculture as live food for the early stage of penaeid larvae

The total ash, chlorophyll, phaeopigment, lipid and fatty acid contents of three microalgae (the diatomsChaetoceros calcitrans, Skeletonema costatum and the prymnesiophyteIsochrysis galbana) used in tropical Malaysian penaeid mariculture were studied. The axenic laboratory cultures were grown in f-2 medium, while outdoor cultures were grown in a commercial medium designed for optimum nutrition in tropical outdoor aquaculture operations. Considerable amounts (3–9% of the total fatty acid) of the polyunsaturated fatty acid 20:5w3 (eicosapentaenoic acid) were present in the diatoms; very small amounts were also found inI. galbana. Lipid contents were three to five times higher than chlorophylla. There was an increase with culture age in the relative proportion of total C18 and C20 fatty acid components. The algae contained the w3-polyunsaturated fatty acids (PUFA) necessary for the growth and survival rate of the prawn larvae. However, onlyChaetoceros calcitrans grew well under the conditions of temperature and salinity in the hatchery.     

Association of insertion/deletion polymorphism of alpha-adrenoceptor gene in essential hypertension with or without type 2 diabetes mellitus in Malaysian subjects

An insertion/deletion (I/D) polymorphism of Alpha2B-Adrenoceptor (ADRA2B) gene located on chromosome 2 has been studied extensively in related to cardiovascular diseases. The main aim of the present study was to examine the potential association of D allele frequency of I/D polymorphism of ADRA2B gene in Malaysian essential hypertensive subjects with or without type 2 diabetes mellitus (T2DM). This study includes 70 hypertensive subjects without T2DM, 65 hypertensive subjects with T2DM and 75 healthy volunteers as control subjects. Genotyping of I/D polymorphism was performed by conventional PCR method. There was significant difference found in age, body mass index, systolic/diastolic blood pressure and high density lipoprotein cholesterol level between the case and control subjects. DD genotypic frequency of I/D polymorphism was significantly higher in hypertensive subjects (42.84% vs. 29.33%; P-=0.029) and in hypertensive with T2DM subjects (46.15% vs. 29.33%; P=0.046) than control group. D allele frequency was higher in hypertensive group (67.41%) than control subjects (52.67%). However, no significant difference was found between the three genotypes of I/D polymorphism of ADRA2B gene and the clinical characteristics of the subjects. The result obtained in this study show D allele of ADRA2B gene was associated with essential hypertension with or without T2DM in Malaysian subjects.     

Improved method for the isolation of RNA from bacteria refractory to disruption, including S. aureus producing biofilm

The development of fast, reliable and inexpensive phenol protocol is described for the isolation of RNA from bacterial biofilm producers. The method was tested on Staphylococcus aureus (S. aureus) and other biofilm-producing gram-negative microorganisms and provided the highest integrity of RNA recovery in comparison to other methods reported here. In parallel experiments, bacterial lysis with Qiagen, NucleoSpin RNAII, InnuREP RNA Mini, Trizol and MasterPure RNA extraction Kits using standard protocols consistently gave low RNA yields with an absence of integrity. The boiling method presented here yielded high concentration of RNA that was free from 16S and 23S rRNA, contained 5S RNA. Higher yields due to improved biofilm bacterial cell lysis were achieved with an added hot phenol incubation step without the need for a bead mill or the enzyme. This method when used in conjunction with the Qiagen RNeasy Mini kit, RNA isolation was a success with greater integrity and contained undegraded 16S and 23S rRNA and did not require further purification. Contaminating DNA was a problem with the RNA processing samples; we used quantitative real-time PCR (RT-qPCR) to measure the recovery of RNA from bacterial biofilm cells using the method described here.