Development of gates to measure the immature platelet fraction in C57BL/6J mice using the Sysmex XN-V series multispecies hematology analyzer

The immature platelet fraction (IPF) is a measure of newly released platelets, which has been used as a marker of platelet production in multiple human studies but is not widely available in multispecies analyzers. We developed gates to measure the IPF in diluted and undiluted murine blood samples on the Sysmex XN-1000V multispecies hematology analyzer. IPF gates were created using undiluted and diluted (1/10) blood samples obtained from adult and newborn (postnatal day 10, P10) C57BL/6J wild-type (WT) mice, and from 3 murine models of thrombocytopenia: c-MPL^−/− mice, which lack the thrombopoietin receptor (hyporegenerative); antibody-mediated thrombocytopenia; and acute inflammation-induced thrombocytopenia. P10 mice were chosen because, at their size, we could consistently obtain (by terminal phlebotomy) the blood volume needed to run an undiluted sample. The undiluted blood IPF gate successfully differentiated between mechanisms of thrombocytopenia in both adult and P10 mice. For diluted samples, 2 IPF gates were generated: a thrombocytopenic (T) gate, which performed well in samples with platelet counts (PCs) <800 × 10⁹/L in adult mice and <500 × 10⁹/L in newborn mice, and a non-thrombocytopenic (NT) gate, which performed well in samples with PCs above these thresholds. PCs and IPFs measured in diluted blood using these gates agreed well with those measured in undiluted blood and had good reproducibility. These diluted gates allow for the accurate measurement of PCs and IPFs in small (10 µL) blood volumes, which can be obtained easily from adult and newborn mice as small as P1 to assess platelet production serially.     

Screening of quinalphos,trifluralin and dichlorvos residues in fresh water of aquaculture systems in Mekong Delta,Vietnam

To develop an easy and reliable method for detecting pesticides and their residues in the Mekong Delta, a GC‐MS analytical method was developed and validated according to European guidelines (SANTE/11945/2015) for the determination of residues of three pesticides (quinalphos, trifluralin and dichlorvos) in water. The limit of detection (LOD) and the limit of quantification (LOQ) were 0.002 and 0.007 μg/L, respectively, for quinalphos and trifluralin, and 0.016 and 0.053 μg/L, respectively, for dichlorvos and quinalphos. The repeatability, the within‐laboratory reproducibility as well as the trueness met the European criteria. The recovery rate ranged between 72% (for dichlorvos and quinalphos) and 82% (for trifluralin). The developed method was then applied for the analysis of 33 water samples, collected in April 2013, at the beginning of the rainy season in the Mekong Delta in Vietnam. Thirteen samples were from rice field, 10 were collected from cat fish ponds and from red tilapia cages. Results showed that only 9% of total water samples analysed contained residues of pesticides, but only in water from rice fish systems. From the 13 samples taken in these systems, quinalphos was detected in three samples. The other two pesticides were not detected. A comparison between analytical results obtained from GC‐MS and an alternative method, that is GC‐ECD indicated that GC‐ECD is less sensitive than GC‐MS, with LOQ ranging from 0.37 to 1.18 (depending on the pesticide). However, for samples with concentrations above these LOQ, no significant difference was observed between the results obtained from the two analytical methodologies.     

Effects of early weaning strategies on growth,survival and digestive enzyme activities in cobia (<Emphasis Type="Italic">Rachycentron canadum</Emphasis> L.) larvae

The effects of weaning strategies of cobia (Rachycentron canadum L.) larvae to commercial microdiets, either from rotifers or from Artemia, on growth, survival and enzymatic digestive capacity, were investigated. In the first experiment, cobia larvae were weaned from rotifers by co-feeding with a microdiet (Otohime) from 8, 13 or 20 days post-hatching (dph). The larvae in the control treatment were fed rotifers (2–12 dph), Artemia nauplii from 7 dph, and co-fed with the microdiet from 20 dph. In the second experiment, the larvae were weaned from Artemia, which was fed to the larvae from 7 dph, by co-feeding with a microdiet (NRD) from 8, 13 or 18 dph. The larvae in control treatment were fed rotifers, then Artemia to the end of the experiment (28 dph). Weaning of cobia larvae onto a microdiet directly from rotifers significantly reduced growth, survival and digestive capacity of the larvae and did not lead to larval acceptance of the microdiet, compared to those weaned from Artemia in the first experiment. Early weaning of cobia larvae onto NRD microdiet (on 8 or 13 dph) from Artemia in the second experiment also reduced growth, survival rate and gut maturation index, compared to those fed live feed. With available microdiets, weaning of cobia larvae could start from Artemia at around 18 dph in order to obtain comparable growth, survival and gut maturation to larvae fed live feed.     

Novel α-Amylase Inhibitor Hemi-Pyocyanin Produced by Microbial Conversion of Chitinous Discards

α-Amylase inhibitors (aAIs) have been applied for the efficient management of type 2 diabetes. The aim of this study was to search for potential aAIs produced by microbial fermentation. Among various bacterial strains, Pseudomonas aeruginosa TUN03 was found to be a potential aAI-producing strain, and shrimp heads powder (SHP) was screened as the most suitable C/N source for fermentation. P. aeruginosa TUN03 exhibited the highest aAIs productivity (3100 U/mL) in the medium containing 1.5% SHP with an initial pH of 7–7.5, and fermentation was performed at 27.5 °C for two days. Further, aAI compounds were investigated for scaled-up production in a 14 L-bioreactor system. The results revealed a high yield (4200 U/mL) in a much shorter fermentation time (12 h) compared to fermentation in flasks. Bioactivity-guided purification resulted in the isolation of one major target compound, identified as hemi-pyocyanin (HPC) via gas chromatography-mass spectrometry and nuclear magnetic resonance. Its purity was analyzed by high-performance liquid chromatography. HPC demonstrated potent α-amylase inhibitory activity comparable to that of acarbose, a commercial antidiabetic drug. Notably, HPC was determined as a new aAI. The docking study indicated that HPC inhibits α-amylase by binding to amino acid Arg421 at the biding site on enzyme α-amylase with good binding energy (−9.3 kcal/mol) and creating two linkages of H-acceptors.     

Classification of Southeast Asian mints (Mentha spp.) based on simple sequence repeat markers

Mentha is a complex genus encompassing many species as a consequence of their interspecific hybridization and polyploidy. Southeast Asian mints have been poorly distinguished though they are widely used for culinary and medical purposes. In this study, we have analyzed Southeast Asian mints and known varieties as well as a related Lamiaceae species (Nepeta sp.) using simple sequence repeat (SSR) markers and leaf morphology. Two types of mints were clearly distinguished based on their venation pattern and leaf shape index. We developed 12 SSR markers that allowed good amplification in the Mentha and another Lamiaceae species. In the SSR-based phylogram, the Mentha lines could be delimited into groups I–VI. The Southeast Asian mints divided into groups I and II, and the phylogram separated most of the available species, with groups I and II containing the known species M. × cordifolia and M. arvensis, respectively. The separation of the two groups was supported by a population structure analysis. The SSR markers developed in this study enabled the simultaneous classification of mints and will help improve our understanding of the genetic composition of known mint varieties and as yet unclassified Southeast Asian mints.     

A closer look into shrimp yields and mangrove coverage ratio in integrated mangrove-shrimp farming systems in Ca Mau,Vietnam

Shrimp aquaculture is the biggest source of export income in Vietnam. However, the development of the shrimp poses a serious threat to coastal mangroves by converting coastal mangroves into shrimp farms. Much effort has been made to replant mangroves and reduce the impacts of shrimp farming on the environment, and maintaining mangrove coverage at 30–50% of total farm area has provided the highest benefits in the integrated mangrove shrimp model. In this study, we re-examine the benefits of forest cover on the survival and yield of tiger shrimp (Penaeus monodon) in the integrated mangrove shrimp farming systems in Ca Mau province, Vietnam. The study found positive linear correlations of log transformed survival and yield of tiger shrimp with forest cover of the forms: Ln (survival)?=????1.39?+?0.038?×?forest coverage (r²?=?0.22; p value?=?0.0007); Ln (yield)?=?3.55?+?0.026?×?forest coverage (r²?=?0.16; p value?=?0.004). The households with high forest cover (i.e.,?>?45%) also had 1.07?±?0.29 and 1.39?±?0.36 (CI 95%, p value?=?0.000) kg ha^?1 higher yield per night harvest than those with medium and low forest cover, respectively. As a result, households with high forest cover have higher benefits than those with lower forest cover. Our findings together with previous published studies lead to the recommendation that farmers maintain mangroves in farming systems for better economic and environmental benefits.

     

Response of laying hens to L-arginine,L-citrulline and guanidinoacetic acid supplementation in reduced protein diet

A study was conducted with Hy-Line Brown laying hens to examine the effects of reduced protein diet, deficiency of arginine (Arg), and addition of crystalline Arg, citrulline (Cit) and guanidinoacetic acid (GAA) as substitutes for Arg. Hen performance, egg quality, serum uric acid, liver and reproductive organ weights, and energy and protein digestibility were measured using a completely randomized design with 5 treatments. Treatments were a standard diet (17% protein diet; SP), a reduced diet (13% protein diet deficient in Arg; RP) and RP with added Arg (0.35%, RP-Arg), GAA (0.46% equivalent to 0.35% Arg, RP-GAA) or Cit (0.35%, RP-Cit) to the level of SP. It was hypothesized that performance would decrease with Arg deficient RP diet and the addition of GAA or Cit in RP would allow birds to perform similar or greater than Arg-added RP treatment. The experiment was conducted from 20 to 39 wk of age but the treatment effect was seen only after 29 wk of age. The birds offered RP had reduced egg and albumin weights (P < 0.01), lower yolk color score (P < 0.01), lower protein intake and excretion (P < 0.01) than those offered SP. When Arg or Cit were added to RP to make them equivalent to SP, feed intake (FI) and egg production were not different than those of RP (P > 0.05). The birds offered RP-GAA decreased FI and egg production (P < 0.01) compared to those offered RP. The addition of Arg, Cit or GAA to the RP had no effect on egg quality parameters, protein and energy digestibilities (P > 0.05). However, birds offered the RP-Cit diet tended to have higher Haugh unit (P = 0.095) and lower shell breaking strength (P = 0.088) compared to all other treatments while those offered RP-GAA had higher energy digestibility (P < 0.05) than all other groups but RP. The limited performance response of hens fed RP with added Arg, GAA, or Cit may be due to deficiency of some other nutrients in RP such as phenylalanine, potassium or non-essential amino acids and other components of soybean meal in the diet.     

Phylogeography and conservation gaps of Musa balbisiana Colla genetic diversity revealed by microsatellite markers

Genetic Resources and Crop Evolution - Collection and storage of crop wild relative (CWR) germplasm is crucial for preserving species genetic diversity and crop improvement. Nevertheless, much of...