Survey of veterinery [sic] drug residues, environmental contaminants and mycotoxins of cultured shirmp [sic] in Iran

Economical achievement of optimal growth in developing countries may lead to sustainable poverty reduction. Agricultural activities play an important role in economy and human being welfare, which leads to establishment of food security and quality. Aquaculture products in developing countries share 51.4 percent of total agricultural production.7—percent in developed countries. Therefore undoutedly food production by means of quality and quantity has to be increased .The history of shirmp production goes back to 500 years ago. Today 50 countries of the world produce shirmp .In Islamic Republic of Iran shrimp production started since 1992 in the coastal region of Persian Gulf. The shrimp culture farms canbe classified in to 4 different categories; Extensive, semi-extensive, intensive and super instensive. Global ecological manitenanc is one of the major concerns of authorities Human manipulation of nature is the most destructive activity. Industrial sweage leakage in to the rivers and water sources is a big issue that causes reduction in the aquatic population. Heavy metals have an inhibitory effect in the production and growth of sealife. Human intake of food treated with anti microbial cause's allergy, hypersensitivity and develops microbial resistance. Organochlorine compounds contamination may found in hepato pancreatic tissues of aquatic products, Aresnic may transfer to man via plant & animal product contamination. In 1991 during Persian Gulf Mir 700 oil well set

Population growth and physiological characteristics of microalgae in a miniaturized bioreactor during space flight

A strain of microalgae (Anabaena siamensis) had been cultured in a miniaturized bioreactor during a retrievable satellite flight for 15 days. By means of remote sensing equipment installed in the satellite, we gained the growth curve of microalgae population in space every day in real time. The curve indicated that the growth of microalgae in space was slower than the control on ground. Inoculation of the retrieved microalgae culture showed that the growth rate was distinctively higher than ground control. But after several generations, both cultures indicated similar growth rates. Those data showed that algae, can adapt to space environment easily which may be valuable for designing more complex bioreactor and controlled ecological life support system in future experiment. (C) 2006 Elsevier Ltd. All rights reserved.

Population genetics of Crassostrea ariakensis in Asia inferred from microsatellite markers

Crassostrea ariakensis is an important aquacultured oyster species in Asia, its native region. During the past decade, consideration was given to introducing C. ariakensis into Chesapeake Bay, in the United States, to help revive the declining native oyster industry and bolster the local ecosystem. Little is known about the ecology and biology of this species in Asia due to confusion with nomenclature and difficulty in accurately identifying the species of wild populations in their natural environment. Even less research has been done on the population genetics of native populations of C. ariakensis in Asia. We examined the magnitude and pattern of genetic differentiation among 10 wild populations of C. ariakensis from its confirmed distribution range using eight polymorphic microsatellite markers. Results showed a small but significant global theta (ST) (0.018), indicating genetic heterogeneity among populations. Eight genetically distinct populations were further distinguished based on population pairwise theta (ST) comparisons, including one in Japan, four in China, and three populations along the coast of South Korea. A significant positive association was detected between genetic and geographic distances among populations, suggesting a genetic pattern of isolation by distance. This research represents a novel observation on wild genetic population structuring in a coastal bivalve species along the coast of the northwest Pacific.

Production of a base population and its responses to F-1 selection in the bay scallop, Argopecten irradians irradians Lamarck (1819)

A base population of the bay scallop, Argopecten irradians irradians Lamarck, was produced by crossing two cultured bay scallop populations. After 1 year of rearing, the top 10% truncation selection of the top 10% (i=1.755) was carried out in the base population of about 1300 adults. A control parental group with a an identical number to the select parental group was randomly selected from the entire population before isolation of the select parental group. The result showed that, at the larval stage, the growth rate of larvae in the selected line was significantly higher than that of the control (P < 0.05), and that the genetic gain was 6.78%. Owing to the lower density of control at the spat stage, the mean shell length of the control line was larger than that of the select line at day 100. When the same density was adjusted between two lines in the grow-out stage (from day 100 to 160), the daily growth rate of the selected line was significantly higher than that of the control line (P < 0.05). Survival of the select line was significantly larger than that of the control line in the grow-out stage. In conclusion, the results obtained from this experiment indicate that selective breeding from a base population with a high genetic diversity established by mass spawning between different populations appears to be a promising method of genetic improvement in bay scallop, A. irradians irradians Lamarck.

Human marrow mesenchymal stem Cells (MSCs) in the peplated non-adherent cell population serve as a complementary source of osteogenic cells

To obtain enough quantity of osteogenic cells is a challenge for successful cell therapy in bone defect treatment, and cell numbers were usually achieved by culturing bone marrow cells in a relatively long duration. This study reported a simple and cost effective method to enhance the number of MSCs by collecting and replating the non-adherent cell population of marrow MSCs culture. Bone marrow MSCs were isolated from 11 patients, cultured at a density of 1×105/cm2 to 1×106/cm2 in flasks. For the first three times of media change, the floating cells were centrifuged and replated in separate flasks. The total number of cells in both the primary and replating flasks were counted at day 21. Cell proliferation rate, potentials for osteogenic, chondrognenic, and adipogenic differentiation were examined in both cell types in vitro. In-vivo osteogenic potentials of the cells were also tested in mice implantation model. The results showed that MSCs derived from non-adherent cell population of marrow cell cultures have similar cell proliferation and differentiation potentials as the originally attached MSCs in vitro. When implanted with HA-TCP materials subcutaneously in SCID mice, newly formed bony tissues were found in both cell type groups with osteocalcin expression. We have obtained 36.6% (20.70%-44.97%) more MSCs in the same culture period when the non-adherent cell populations were collected. The findings confirmed that the non-adherent cell population in the bone marrow culture is a complementary source of MSCs, collecting these cells is a simple and cost-effective way to increase MSCs numbers and reduce the time required for culturing MSCs for clinical applications.

A novel mutation in the sulfate transporter gene SLC26A2 (DTDST) specific to the Finnish population causes de la Chapelle dysplasia.

BACKGROUND: Mutations in the sulfate transporter gene SLC26A2 (DTDST) cause a continuum of skeletal dysplasia phenotypes that includes achondrogenesis type 1B (ACG1B), atelosteogenesis type 2 (AO2), diastrophic dysplasia (DTD), and recessive multiple epiphyseal dysplasia (rMED). In 1972, de la Chapelle et al reported two siblings with a lethal skeletal dysplasia, which was denoted "neonatal osseous dysplasia" and "de la Chapelle dysplasia" (DLCD). It was suggested that DLCD might be part of the SLC26A2 spectrum of phenotypes, both because of the Finnish origin of the original family and of radiographic similarities to ACG1B and AO2. OBJECTIVE: To test the hypothesis whether SLC26A2 mutations are responsible for DLCD. METHODS: We studied the DNA from the original DLCD family and from seven Finnish DTD patients in whom we had identified only one copy of IVS1+2T>C, the common Finnish mutation. A novel SLC26A2 mutation was found in all subjects, inserted by site-directed mutagenesis in a vector harbouring the SLC26A2 cDNA, and expressed in sulfate transport deficient Chinese hamster ovary (CHO) cells to measure sulfate uptake activity. RESULTS: We identified a hitherto undescribed SLC26A2 mutation, T512K, homozygous in the affected subjects and heterozygous in both parents and in the unaffected sister. T512K was then identified as second pathogenic allele in the seven Finnish DTD subjects. Expression studies confirmed pathogenicity. CONCLUSIONS: DLCD is indeed allelic to the other SLC26A2 disorders. T512K is a second rare "Finnish" mutation that results in DLCD at homozygosity and in DTD when compounded with the milder, common Finnish mutation.

Experimental evidence and model explanation for cell population characteristics modification when applying sequential photodynamic therapy

We present experimental evidence of the existence of cell variability in terms of threshold light dose for Hep G2 (liver cancer cells) cultured. Using a theoretical model to describe the effects caused by successive photodynamic therapy (PDT) sessions, and based on the consequences of a partial response we introduce the threshold dose distribution concept within a tumor. The experimental model consists in a stack of flasks, and simulates subsequent layers of a tissue exposed to PDT application. The result indicates that cells from the same culture could respond in different ways to similar PDT induced-damages. Moreover, the consequence is a partial killing of the cells submitted to PDT, and the death fraction decreased at each in vitro PDT session. To demonstrate the occurrence of cell population modification as a response to PDT, we constructed a simple theoretical model and assumed that the threshold dose distribution for a cell population of a tumor is represented by a modified Gaussian distribution.

Fresh and cultured buccal cells as a source of mRNA and protein for molecular analysis

We developed a method for obtaining viable buccal cells from mouthwash samples for use as a source of mRNA and protein. Immunofluorescent analysis showed that most cells were derived from nonkeratinized parabasal epithelia, with a minor proportion of proliferative cells. Gene expression was detected in buccal cells using reverse transcription PCR, Western blot analysis, and immunofluorescence. Using a keratinocyte-specific medium, buccal cells could be cultured on Matrigel™-coated permeable filters for up to 2 weeks while maintaining the expression of some epithelial-specific markers, including cytokeratin 13, cytokeratin 10, transferrin receptor, and β-integrin. The basal marker cytokeratin 14 and Ki67, an indicator of cellular proliferation, were detected in a few cells. We show that buccal cells can be obtained from a noninvasive procedure for use as a source of material for biochemical analyses. A population of the buccal cells can be maintained in culture for up to 2 weeks using keratinocyte-specific medium in combination with extracellular matrix.

A rapid cell counting method utilising acridine orange as a novel discriminating marker for both cultured astrocytes and microglia

Cell culture analyses of growth, morphology and apoptosis commonly require counting of different cell types stained with antibodies to discriminate between them. Previously, we reported the use of l-Leucine methyl ester (l-LME) to prepare purified cultures of type 1 astrocytes with minimal microglia, and staining by GFAP and CD antibodies, respectively. Here, we demonstrate a novel use of acridine orange (AO) for rapid discrimination between these cell types using fluorescence microscopy. AO accumulates in the lysosomes and also binds strongly to nuclear DNA and cytoplasmic/nucleolar RNA. Microglia may contain abundant lysosomes due to known roles in homeostasis and immune response. AO staining of lysosomes was tested at a range of concentrations, and 2.5 μg/mL was most suitable. In agreement with previous reports, microglia treated with AO showed very intense yellow, orange or red granular cytoplasmic staining of lysosomes. Microglia contain a substantially higher number of lysosomes than astrocytes, which have a variable amount. We measured the microglia population at 5.14 ± 0.50% in mixed cultures. Thus, these results show AO is a novel discriminatory marker, as microglia were easily observed and counted in clumps on top of the monolayer of astrocytes, providing a rapid alternative to time-consuming and costly antibody-based assays.

Mitochondrial DNA diversity of broodstock of two indigenous mahseer species, Tor tambroides and T. douronensis (Cyprinidae) cultured in Sarawak, Malaysia

Tor tambroides and T. douronensis, locally referred to as empurau and semah, respectively, are high valued mahseer species, indigenous to Sarawak, East Malaysia, with an aquaculture potential and of conservational value. Direct sequencing of mitochondrial DNA (mtDNA) 16S rRNA gene region (542 bp) was used to investigate genetic variation of T. tambroides and T. douronensis broodstock collected from different geographic locations in Sarawak and maintained at the Indigenous Fish Research and Production Center (IFRPC), Tarat, Sarawak, Malaysia. A total of 11 unique haplotypes were identified, of which six were detected in T. tambroides, and five in T. douronensis. Overall, nucleotide diversity (π) was low, ranging from 0.000 to 0.006, and haplotype diversity (h) ranged from 0.000 to 0.599. Although the analysis failed to detect genetic variation amongst populations of T. tambroides (significant pairwise FST was found for only one test, but pairwise haplotype frequencies were not statistically significant), substantial inter-population divergence among T. douronensis was recognised, especially those originating from different river systems (pairwise F_ST = 0.754 to 1.000, P < 0.05). Fixed haplotype differences were found in one population of T. douronensis. Average nucleotide divergence between T. tambroides and T. douronensis was 0.018, similar to the amount recognised between T. tambroides and the outgroup T. khudree (0.017). In addition, phylogenetic analysis revealed that the T. douronensis mtDNA consisted of two highly divergent clusters (0.020), one of which is more closely related to T. tambroides rather than with the other group of haplotypes of the conspecifics. The findings from the present study have important implications for aquaculture, management and conservation of these two species. The data also raise some concerns regarding the taxonomic status of T. douronensis, which needs to be addressed.

Population structure and life history characteristics of euastacus bispinosus and cherax destructor (Parastacidae) in the Grampians National Park, Australia

Despite being the most widely distributed and cultured freshwater crayfish species in Australia, relatively little is known about the ecology of Cherax destructor outside of captivity. Similarly, few ecological studies have been conducted on the large and threatened stream dwelling Australian freshwater crayfish Euastacus bispinosus. A series of seasonal sampling surveys over two years investigated the population structure, life history and reproductive timing of C. destructor in fire dam habitats, and of E. bispinosus in channel (stream) habitats, in the Grampians National Park in south-west Victoria, Australia. Cherax destructor individuals in the largest size class (50 – 59.95 mm OCL) were not abundant during the study, while those belonging to the 0 – 9.95 mm OCL size class were more frequent in summer than at other times of year, suggesting synchronous recruitment in fire dam habitats. Individuals in the reproductively active mid-size classes were also frequent in summer. For E. bispinosus, sex ratios in spring always favoured females, although there were no clear trends for other times of year. Gravid E. bispinosus females were found in winter and spring throughout the study, and were sexually mature at a smaller size than has previously been reported.

Population trends of the red palm mite, raoiella indica hirst (acari: tenuipalpidae) and associated entomopathogenic fungi in trinidad, antigua, st kitts and nevis and dominica

A survey to determine population trends and entomopathogenic fungi associated with the red palm mite (RPM), Raoiella indica, was conducted in Trinidad, Antigua, St. Kitts and Nevis and Dominica. RPM population density was evaluated by sampling a total of ten coconut palms per site in Antigua, St. Kitts and Nevis, Dominica, and Trinidad (Manzanilla and Icacos). Mites from the four islands were either surface sterilized or left unsterilized before being cultured on Tap Water Agar (TWA). A total of 318 fungal colonies were retrieved. A further 96 mites from Dominica were kept on sterile moist filter paper in a humidity chamber and a further 85 colonies were isolated. Based on morphological observations of all 403 isolates, a sample consisting of 32 colonies (8 %) was sent for identification at CABI-UK. Of the 27 fungi positively identified, 15 isolates belonged to the genera Cladosporium, three to Simplicillium spp., and one to Penicillium. Other fungi genera with limited or no entomopathogenic potential included: Aspergillus, Cochliobolus, Fusarium, Pestalotiopsis and Pithomyces. The results show a potential use of entomopathogenic fungi for population management of the red palm mite in the Caribbean region.

XIPHOPHORUS ENZYME GENE EXPRESSION: TISSUE SPECIFICITY, GENETIC CONTROL, AND STABILITY IN CULTURED CELLS

Five permanent cell lines were developed from Xiphophorus maculatus, X. helleri, and their hybrids using three tissue sources, including adults and embryos of different stages. To evaluate cell line gene expression for retention of either tissue-of-origin-specific or ontogenetic stage-specific characters, the activity distribution of 44 enzyme loci was determined in 11 X. maculatus tissues, and the developmental genetics of 17 enzyme loci was charted in X. helleri and in helleri x maculatus hybrids using starch gel electrophoresis. In the process, eight new loci were discovered and characterized for Xiphophorus.^ No Xiphophorus cell line showed retention of tissue-of-origin-specific or ontogenetic stage-specific enzyme gene expressional traits. Instead, gene expression was similar among the cell lines. One enzyme, adenosine deaminase (ADA) was an exception. Two adult-origin cell lines expressed ADA, whereas, three cell lines derived independently from embryos did not. ADA('-) expression of Xiphophorus embryo-derived cell lines may represent retention of an embryonic gene expressional trait. In one cell line (T(,3)) derived from 13 pooled interspecific hybrid (F(,2)) embryos, shifts with time were observed at enzyme loci polymorphic between the two species. This suggested shifts in ratios of cells of different genotypes in the population rather than unstable gene expression in one dominant cell type.^ Verification of this hypothesis was attempted by cloning the culture--seeking clones having different genetic signatures. The large number of loci electrophoretically polymorphic between the two species and whose alleles segregated independently into the 13 progeny from which this culture originated almost guaranteed the presence of different genetic signatures (lineages) in T(,3).^ Seven lineages of cells were found within T(,3), each expressing genotypes at some loci not characteristic of the expression of the culture-as-a-whole, supporting the hypothesis tested. Quantitative studies of ADA expression in the whole culture (ADA('-)) and in clones of these seven lineages suggested the predominance in T(,3) of ADA deficient cell lineages, although moderate to high ADA output clones also occurred. Thus, T(,3) has the potential to shift phenotypes from ADA('-) to ADA('+) by simply changing proportions of its constituent cell types, demonstrating that such shifts can occur in any cell culture containing cells of mixed expressional characteristics.^

(Table 1) Population density and biomass of the foraminifer Rupertina, and grain size of cores from the Norwegian continental slope

Rupertina stabilis occupies a depth restricted biotope of suspension feeding animals situated at the Norwegian continental margin. It extends from the Voring plateau northwards for at least 200 - 300 km, in depths between 600 and 800 m. This slope position is known for relatively strong bottom currents and shifting watermass boundaries. - The species is attached to hard substrates, mainly stones or hydroid stalks and obviously prefers an elevated position. It is building a permanent cyst of sponge spicules and debris at the apertural region. The spicules are used to support a pseudopodial network similar to that described from Halyphysema (LIPPS 1983). It is believed to serve as a filter apparatus. - A review of known occurences in the Atlantic is given, suggesting a temperature adaption of the species ranging from 0°C to a maximum of 8°C. Specimens were successfully cultured for about 2-3 weeks.

Multilocus analysis of extracellular putative virulence proteins made by group A Streptococcus: Population genetics, human serologic response, and gene transcription

Species of pathogenic microbes are composed of an array of evolutionarily distinct chromosomal genotypes characterized by diversity in gene content and sequence (allelic variation). The occurrence of substantial genetic diversity has hindered progress in developing a comprehensive understanding of the molecular basis of virulence and new therapeutics such as vaccines. To provide new information that bears on these issues, 11 genes encoding extracellular proteins in the human bacterial pathogen group A Streptococcus identified by analysis of four genomes were studied. Eight of the 11 genes encode proteins with a LPXTG(L) motif that covalently links Gram-positive virulence factors to the bacterial cell surface. Sequence analysis of the 11 genes in 37 geographically and phylogenetically diverse group A Streptococcus strains cultured from patients with different infection types found that recent horizontal gene transfer has contributed substantially to chromosomal diversity. Regions of the inferred proteins likely to interact with the host were identified by molecular population genetic analysis, and Western immunoblot analysis with sera from infected patients confirmed that they were antigenic. Real-time reverse transcriptase–PCR (TaqMan) assays found that transcription of six of the 11 genes was substantially up-regulated in the stationary phase. In addition, transcription of many genes was influenced by the covR and mga trans-acting gene regulatory loci. Multilocus investigation of putative virulence genes by the integrated approach described herein provides an important strategy to aid microbial pathogenesis research and rapidly identify new targets for therapeutics research.