Heterologous expression and characterization of an N-acetyl-β-D-hexosaminidase from Lactococcus lactis ssp. lactis IL1403

The lnbA gene of Lactococcus lactis ssp. lactis IL1403 encodes a polypeptide with similarity to lacto-N-biosidases and N-acetyl-β-D-hexosaminidases. The gene was cloned into the expression vector pET-21d and overexpressed in Escherichia coli BL21* (DE3). The recombinant purified enzyme (LnbA) was a monomer with a molecular weight of approximately 37 kDa. Studies with chromogenic substrates including p-nitrophenyl N-acetyl-β-D-glucosamine (pNP-GlcNAc) and p-nitrophenyl N-acetyl-β-D-galactosamine (pNP-GalNAc) showed that the enzyme had both N-acetyl-β-D-glucosaminidase and N-acetyl-β-D-galactosaminidase activity, thus indicating that the enzyme is an N-acetyl-β-D-hexosaminidase. K(m) and k(cat) for pNP-GlcNAc were 2.56 mM and 26.7 s(-1), respectively, whereas kinetic parameters for pNP-GalNAc could not be determined due to the K(m) being very high (>10 mM). The optimal temperature and pH of the enzyme were 37 °C and 5.5, respectively, for both substrates. The half-life of activity at 37 °C and pH 6.0 was 53 h, but activity was completely abolished after 30 min at 50 °C, meaning that the enzyme has relatively low temperature stability. The enzyme was stable in the pH 5.5-8 range and was unstable at pH below 5.5. Studies with natural substrates showed hydrolytic activity on chito-oligosaccharides but not on colloidal chitin or chitosan. Transglycosylation products were not detected. In all, the data suggest that LnbA's role may be to degrade chito-oligosaccharides that are produced by the previously described chitinolytic system of L. lactis.     

Differentiation of Umbilical Cord Lining Membrane-Derived Mesenchymal Stem Cells into Endothelial-Like Cells

Background: Stem cell therapy for the treatment of vascular-related diseases through functional revascularization is one of the most important research areas in tissue engineering. The aim of this study was to investigate the in vitro differentiation of umbilical CL-MSC into endothelial lineage cells. Methods: In this study, isolated cells were characterized for expression of MSC-specific markers and osteogenic and adipogenic differentiation. They were induced to differentiate into endothelial-like cells and then examined for expression of the endothelial-specific markers, karyotype, and functional behavior of cells. Results: Isolated cells expressed MSC-specific markers and differentiated into adipocytes and osteoblasts. After endothelial differentiation, they expressed CD31, vWF, VE-cadherin, VEGFR1, and VEGFR2 at both mRNA and protein level, but their morphological changes were not apparent when compared with those of undifferentiated cells. There were no significant changes in karyotype of differentiated cells. Furthermore, angiogenesis assay and LDL uptake assay showed that differentiated cells were able to form the capillary-like structures and uptake LDL, respectively. Conclusion: The results indicated that umbilical CL-MSC could differentiate into functional endothelial-like cells. Also, they are suitable for basic and clinical studies to cure several vascular-related diseases. Key Words: Endothelial differentiation, Endothelial-like cells, Mesenchymal stem cells, Umbilical cord lining membrane     

竹集成材高频热压过程中板坯内温度的变化趋势

以毛竹Phyllostachys edulis集成材为研究对象,在不同条件下进行高频热压,通过对竹集成材板坯高频热压过程中芯层温度变化的统计分析,得到了竹集成材高频热压过程中板坯温度的变化规律。结果表明:在试验条件范围内,随着板坯含水率从6%增加到18%,涂胶量从200 g·m-2增加到300 g·m-2,板坯的温度明显升高。升温过程可以分为快速升温和慢速升温2个阶段。在快速升温阶段板坯内的温度随板坯初含水率、涂胶量及加热时间的提高而递增;在慢速升温阶段板坯初含水率及涂胶量对板坯内的温度的影响很小,板坯芯层升温速度随加热时间的增加而减少。通过试验数据分析,得出较优的高频热压胶合工艺条件为:涂胶量300 g·m-2,竹条含水率12%,高频热压时间7 min。     

Optimization of in vitro embryo production and zygote vitrification for the indigenous Vietnamese Ban pig: The effects of different in vitro oocyte maturation systems

The Vietnamese Ban pig is a precious genetic resource that needs to be preserved. In vitro embryo production from in vitro matured (IVM) oocytes is an important tool for the utilization of cryopreserved porcine sperm. The aim of this study was to compare two media for the IVM of Ban pig oocytes. Immature oocytes were subjected to IVM either in a non‐defined (TCM‐199 + pig follicular fluid) or in a defined base medium (POM + epidermal growth factor). At the end of IVM, the oocytes were in vitro fertilized (IVF) with frozen Ban sperm. Ten hours after IVF, the oocytes were either subjected to orcein staining to check fertilization and maturation status or cultured in vitro for 7 days. There was no difference between the two IVM media in terms of percentages of oocyte maturation and blastocyst production. However, the percentage of male pronuclear formation after IVF and the total cell numbers in blastocysts were higher with the defined system. Zygotes obtained by the two IVM systems survived vitrification at similar rates. In conclusion, the two IVM systems were both effective for the production of Ban pig embryos; however, better embryo quality was achieved with the defined one.     

Assessment of the water quality of two rivers in Hanoi City and its suitability for irrigation water

The To Lich and Kim Nguu Rivers in Hanoi City are the main sources of irrigation water for suburban agricultural land and fish farm. Wastewater from the industrial plants located along these rivers has been discharged, and has degraded the water quality of the rivers. This study describes the chemical properties of water from the rivers, focusing on heavy metal pollution and the suitability of water quality for irrigation water. Water from the rivers was heavily polluted with organic matter and heavy metals such as Pb, Cu, Zn, Cr, Cd and Ni. Dissolved oxygen, chemical oxygen deman, and total suspended solids, and the concentrations of all heavy metals exceeded the Vietnamese standard for surface water quality in all investigated sites. The concentrations of some heavy metals such as Cu, Cd, Cr and Ni were above the internationally recommended WHO maximum level for irrigation water. A wide variation in the heavy metal concentration of water due to metal types is the result of wastewater discharged from different industrial sources.     

Dominant sources of dietary carbon and nitrogen for shrimp reared in extensive rice-shrimp ponds

Stable isotope analysis was used to determine the sources of dietary nitrogen (N) and carbon (C) for shrimp during the rearing phase in extensive rice‐shrimp ponds in My Xuyen and Gia Rai districts, Vietnam. Farm‐made feed was added as a food source in shrimp ponds in My Xuyen district, and based on stable isotope analyses, was generally a poor dietary source. The commercial formulated feed used in Gia Rai also appeared to contribute little directly to the nutritional needs of the shrimp. In contrast, the natural biota in all ponds appeared to contribute substantially. In particular, biota from beam trawls and benthic organic matter were the most likely sources of nutrition in My Xuyen ponds, while benthic organic matter was the main source in Gia Rai ponds. δ¹⁵N ratios in the natural biota in My Xuyen farms decreased over the growing season, suggesting increased N fixation in the case of the benthic organic matter reaching values as low as 1‰. This suggests N‐limitation in the ponds and that natural biota become increasingly dependent on N fixed by algae and/or other microorganisms. There is the potential to promote the growth of the plankton and hence, the other natural biota, by the judicious addition of fertilizer.     

Effects of Integrated Shrimp Farming in Vietnam

Despite being a leading shrimp exporter in the world market, shrimp farming in Vietnam is carried out by many small‐scale farmers who face constraints in complying with increasingly stringent food safety and sustainability standards. The best way for such small farmers to circumvent this paradox is to make effective integrations both horizontally and vertically. Using value chain and institutional economics approaches, this study determines the factors that influence the probability of horizontal and vertical integrations between the production and processing stages in the shrimp industry in Vietnam. Analysis results confirm that integrated shrimp aquaculture is much more effective than the traditionally small‐scale‐ and individual‐production mode. In addition, logit estimation results identify the key factors affecting integration, including effectiveness of the legal framework, production scale, infrastructure development, and farmers' skill and awareness. Several policy recommendations proposed to encourage the development of integration in shrimp farming in Vietnam could also be useful for other similar developing countries.     

Sperm Motilities in Waigieu Seaperch,Psammoperca waigiensis: Effects of Various Dilutions,pH, Temperature,Osmolality, and Cations

The objectives of this study were to evaluate the effects of varying dilutions, pH, temperature, osmolality, and cations on sperm motility parameters in waigieu seaperch, Psammoperca waigiensis. The maximum velocity of average in path (VAP), percentage of motile cells (MOT), and duration of sperm motility (DSM) were observed when semen was diluted in artificial seawater (ASW) at a ratio of 1:100 (144.9 ± 0.6 µm/sec, 95.6 ± 0.4%, and 230.3 ± 2.3 sec, respectively), at 30 C (142.0 ± 0.6 µm/sec, 93.6 ± 0.4%, and 238.3 ± 0.9 sec, respectively), and pH 8 (144.8 ± 0.6 µm/sec, 93.3 ± 0.4%, and 234.0 ± 1.5 sec, respectively). Maximum VAP, MOT, and DSM were obtained in each solution containing 0.6 M NaCl (143.8 ± 1.0 µm/sec, 91.3 ± 2.0%, and 230.6 ± 4.2 sec, respectively), 0.6 M KCl (135.1 ± 3.1 µm/sec, 91.1 ± 3.1%, and 230.3 ± 3.7sec, respectively), 0.2M CaCl₂ (105.3 ± 4.7μm/sec, 47.9 ± 2.7%, and 120.7 ± 1.3 sec, respectively), 0.2 M MgCl₂ (107.3 ± 3.0 m/s, 42.1 ± 3.3%, and 120.3 ± 4.8 sec, respectively), and osmolality of 400 mOsm/kg (145.1 ± 2.5 µm/sec, 93.0 ± 2.1%, and 346.5 ± 4.4s, respectively). We used these mediums as artificial insemination media for fertilizing matured eggs. The results showed that the fertilization and hatching rates in 0.6 M NaCl (75.3 ± 0.6% and 57.0 ± 2.4%, respectively), ASW (70.8 ± 1.2% and 51.2 ± 1.8%, respectively), or 400 mOsm/kg (72.9 ± 1.8% and 55.3 ± 1.6%, respectively) were higher than that in seawater (63.9 ± 1.2% and 39.2 ± 3.9%, respectively). In conclusion, using 0.6 M NaCl, ASW, or 400 mOsm/kg as an artificial insemination medium is effective for fertilizing of waigieu seaperch.     

Genetic characterization of chicken anemia viruses newly isolated from diseased chicks in Japan in 2020

In this study, a total of nine chicken samples obtained from two broiler flocks in Oita and Tottori prefectures in 2020 were examined for Chicken anemia virus (CAV) infection. The samples were collected from clinically suspected flocks and diseased chickens. The CAV genome was detected in all nine samples tested by real-time PCR. Phylogenetic analyses and sequence comparisons of the full-length VP1 gene sequences indicated that all the Japanese CAV strains obtained in this study formed a similar cluster of genotype III and shared high nucleotide (99.62–100%) identity. The current Japanese CAV strains were closely related to Chinese CAV strains but not related to vaccine strains. One positive selection site of VP1 was detected among the Japanese CAV strains.     

Occurrence of tetrodotoxin in three Nassarius gastropod species in Khanh Hoa Province,Vietnam

Fisheries Science - Neurotoxic poisonings with fatal symptoms caused by consumption of Nassarius gastropods have been reported in Vietnam but the causative toxins have not been confirmed. In the...