Complementary Analytical Platforms of NMR Spectroscopy and LCMS Analysis in the Metabolite Profiling of Isochrysis galbana

This study was designed to profile the metabolites of Isochrysis galbana, an indigenous and less explored microalgae species. ¹H Nuclear Magnetic Resonance (NMR) spectroscopy and Liquid Chromatography-Mass Spectrometry (LCMS) were used to establish the metabolite profiles of five different extracts of this microalga, which are hexane (Hex), ethyl acetate (EtOAc), absolute ethanol (EtOH), EtOH:water 1:1 (AqE), and 100% water (Aq). Partial least square discriminant analysis (PLS–DA) of the generated profiles revealed that EtOAc and Aq extracts contain a diverse range of metabolites as compared to the other extracts with a total of twenty-one metabolites, comprising carotenoids, polyunsaturated fatty acids, and amino acids, that were putatively identified from the NMR spectra. Meanwhile, thirty-two metabolites were successfully annotated from the LCMS/MS data, ten of which (palmitic acid, oleic acid, α-linolenic acid, arachidic acid, cholesterol, DHA, DPA, fucoxanthin, astaxanthin, and pheophytin) were similar to those present in the NMR profile. Another eleven glycerophospholipids were discovered using MS/MS-based molecular network (MN) platform. The results of this study, besides providing a better understanding of I. galbana’s chemical make-up, will be of importance in exploring this species potential as a feed ingredient in the aquaculture industry.     

Effects of dietary sodium selenite and selenium yeast on antioxidant enzyme activities and oxidative stability of chicken breast meat

The effects of sodium selenite (SS) and selenium yeast (SY) alone and in combination (MS) on the selenium (Se) content, antioxidant enzyme activities (AEA), total antioxidant capacity (TAC), and oxidative stability of chicken breast meat were investigated. The results showed that the highest (p < 0.05) glutathione peroxidase (GSH-Px) activity was found in the SS-supplemented chicken breast meat; however, SY and MS treatments significantly increased (p < 0.05) the Se content and the activities of catalase (CAT), total superoxide dismutase (T-SOD), and TAC, but decreased (p < 0.05) the malondialdehyde (MDA) content at 42 days of age. Twelve days of storage at 4 °C decreased (p < 0.05) the activity of the GSH-Px, but CAT, T-SOD, and TAC remained stable. SY decreased the lipid oxidation more effectively in chicken breast meat. It was concluded that SY and MS are more effective than SS in increasing the AEA, TAC, and oxidative stability of chicken breast meat.     

转基因育种技术在菊花性状改良中的应用进展

 自1989 年发现了菊花对农杆菌浸染的敏感性,农杆菌介导转化法已成功应用于菊花花色、花期、株形、抗病虫及耐非生物胁迫等性状的改良中。从影响菊花遗传转化效率的农杆菌菌株类型、外植体选用、辅助添加剂的使用以及抗生素筛选等几方面阐述了菊花遗传转化技术研究的进展和应用现状,对菊花分子育种性状改良和重要功能基因发掘等研究进展进行综述,并对菊花转化效率改善途径的探索和环境友好型转基因新技术研发前景提出展望,以期为菊花分子育种提供参考。     

Combining Phytate/Ca2+ Fractionation with Trichloroacetic Acid/Acetone Precipitation Improved Separation of Low-Abundant Proteins of Wheat (Triticum aestivum L.) Leaf for Proteomic Analysis

Proteomic assessment of low-abundance leaf proteins is hindered by the large quantity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) present within plant leaf tissues. In the present study, total proteins were extracted from wheat (Triticum aestivum L.) leaves by a conventional trichloroacetic acid (TCA)/acetone method and a protocol first developed in this work. Phytate/Ca²⁺ fractionation and TCA/acetone precipitation were combined to design an improved TCA/acetone method. The extracted proteins were analysed by two-dimensional gel electrophoresis (2-DE). The resulting 2-DE images were compared to reveal major differences. The results showed that large quantities of Rubisco were deleted from wheat leaf proteins prepared by the improved method. As many as (758±4) protein spots were detected from 2-DE images of protein extracts obtained by the improved method, 130 more than those detected by the TCA/acetone method. Further analysis indicated that more protein spots could be detected at regions of pI 4.00–4.99 and 6.50–7.00 in the improved method-based 2-DE images. Our findings indicated that the improved method is an efficient protein preparation protocol for separating low-abundance proteins in wheat leaf tissues by 2-DE analysis. The proposed protocol is simple, fast, inexpensive and also applicable to protein preparations of other plants.     

The nutritional value of peanut hay (Arachis hypogaea L.) as an alternate forage source for sheep

The aim of this study was to evaluate the nutritional and feeding value of peanut hay (Arachis hypogaea L.) produced under tropical environment as an alternate forage resource for sheep. Peanut hay was appreciably high in crude protein [CP; 105 g/kg dry matter (DM)] and lower in neutral detergent fiber (NDF; 466 g/kg DM). Moreover, peanut hay was rich in Ca (12 g/kg DM) and P (1.7 g/kg DM). A feeding trial was conducted to investigate the effect of substituting wheat straw with peanut hay on nutrient intake, digestibility, and N utilization. Four adult Ramghani (Kaghani × Rambouillet) wethers (60?±?2.5 kg body weight) were randomly assigned to the four dietary treatments according to a 4?×?4 Latin square design. The four rations were formulated on isonitrogenous and isocaloric bases and differed in the proportion (in grams per kilogram DM) of wheat straw/peanut hay, i.e., 700:0, 460:240, 240:460, and 0:700. The replacement of wheat straw with peanut hay increased the intakes of DM (P?<?0.001), NDF (P?<?0.01), and N (P?<?0.001). Moreover, apparent in vivo digestibility of DM, NDF, and CP increased (P?<?0.001) with the increasing proportion of peanut hay in the ration. Nitrogen retention in the body increased (P?<?0.01; 3.2 to 8.1 g/day) with the replacement of wheat straw with peanut hay. These findings showed that substitution of wheat straw with peanut hay can improve DM and nutrients intake, digestibility, and N retention in sheep.     

Synthesis,Characterization, and Antibacterial Activity of Cross-Linked Chitosan-Glutaraldehyde

This present study deals with synthesis, characterization and antibacterial activity of cross-linked chitosan-glutaraldehyde. Results from this study indicated that cross-linked chitosan-glutaraldehyde markedly inhibited the growth of antibiotic-resistant Burkholderia cepacia complex regardless of bacterial species and incubation time while bacterial growth was unaffected by solid chitosan. Furthermore, high temperature treated cross-linked chitosan-glutaraldehyde showed strong antibacterial activity against the selected strain 0901 although the inhibitory effects varied with different temperatures. In addition, physical-chemical and structural characterization revealed that the cross-linking of chitosan with glutaraldehyde resulted in a rougher surface morphology, a characteristic Fourier transform infrared (FTIR) band at 1559 cm⁻¹, a specific X-ray diffraction peak centered at 2θ = 15°, a lower contents of carbon, hydrogen and nitrogen, and a higher stability of glucose units compared to chitosan based on scanning electron microscopic observation, FTIR spectra, X-ray diffraction pattern, as well as elemental and thermo gravimetric analysis. Overall, this study indicated that cross-linked chitosan-glutaraldehyde is promising to be developed as a new antibacterial drug.     

A simple approach for regulating saline groundwater inflows to natural streams in irrigated agricultural areas

Stream salinity management is of prime importance for ensuring environmental sustainability of rivers, streams, lakes and other water bodies. Saline groundwater inflows are the main cause of deteriorating quality of stream flows; especially during periods of low flows. The Murray-Darling Basin is a good example for reducing groundwater recharge from irrigated agricultural areas with successful implementation of land and water management plans (LWMPs). In most cases, these plans help minimize the influence of saline groundwater flows on the stream salinity, as a preventive measure. As a remedial measure, this paper introduces a simple but an innovative approach for regulating saline groundwater inflows to natural streams in irrigated agricultural areas. The approach uses the flap gate with an automatic lowering and raising mechanism. During low stream flow, this gate is kept in a raised position. Depending upon the upstream inflows, the ponding situation will develop hydraulic gradient away from the stream; groundwater inflows to the creek will become minimal. During high stream flows, the gate is kept in a lower position to let the diluted water flows through the stream uninterrupted. To install, operate and manage such gates on multiple sites along the stream(s), the individual control panels can be joined into a central control station via telemetric link up. This approach, which holds a key for successfully regulating groundwater induced salinity to natural streams, should be considered in conjunction with other LWMPs to improve stream salinity in an irrigated agricultural area.     

Immunoprotective role of LaSota vaccine under immunosuppressive conditions in chicken challenged with velogenic avian avulavirus-1

The first objective of the present study was to evaluate if the antibodies induced by the live LaSota and killed Newcastle disease (sub-genotype VIIi) vaccines protect the chickens against exposure with pathogenic avian avulavirus-1 (AAvV-1) of chicken and/or pigeon origins. The second objective was to study the effect of vaccines on stressed birds (dexamethasone, aflatoxin, and heat stressed) with respect to antibody production and protection against pathogenic AAvV-1 challenge. Sixty-one-day-old Hubbard chickens were divided into six groups (gA–gF) with ten animals each. All the groups received LaSota (10⁵ EID₅₀, 0.1 ml per chick) on days 7 and 27 via eye drop and one intramuscular injection of a killed vaccine (sub-genotype VIIi) (10^7.5 EID₅₀, 1 ml) on day 18, except the control birds received the PBS only. Moreover, group gC-DEX received dexamethasone intramuscularly at a dose rate of 1-mg/kg body weight daily; gD-AFLA had received aflatoxin as oral gavage at a dose rate of 30 ppb daily, and gE-HEAT was kept under heat stressed (38 °C) till challenged. All the groups were challenged with AAvV-1 strain of chicken origin of sub-genotype VIIi, except the group gA-pigeon was challenged with pigeon-origin strain (sub-genotype VIm). The result showed that the gA-pigeon and gB-chicken vaccinate showed 100% and 80% protection. The immunosuppressive birds produced low pre-challenge HI titer, and protection was observed at 40%, 50%, and 70% in gC-DEX, gD-AFLA, and gE-HEAT, respectively. Our findings suggest the stress factors such as aflatoxin in the feed and dexamethasone are immunosuppressive in nature and suppress the immune response and its associated protective role during infection.

     

Virulence-associated genes and antimicrobial resistance among avian pathogenic Escherichia coli from colibacillosis affected broilers in Pakistan

Avian pathogenic Escherichia coli (APEC) causes colibacillosis that leads to high morbidity and mortality among poultry birds. To date, there is a lack of knowledge about virulence-associated genes (VAGs) and multidrug resistance of APEC isolates from Pakistan. In this study, we determined the VAGs and antibiotic resistance profiles of APEC isolates recovered from colibacillosis affected broilers in Faisalabad region of Pakistan. A total of 84 diseased and dead birds from different local broilers farms were collected and examined for the gross lesions of colibacillosis by conducting postmortem examination. Of these, APEC isolates were recovered from 75 (89.2%) birds. Antibiotic susceptibility tests against 11 antimicrobial agents showed the highest resistance against ampicillin (98.6%) followed by tetracycline (97.3%) and ciprofloxacin (72%). The presence of 11 virulence-associated genes (VAGs) was detected by multiplex polymerase chain reaction (PCR). Of the 75 APEC, 32 (42.6%) harbored > 5 VAGs. Most commonly found genes were increased serum survival (iss; 84%), iron transport (iutA; 74.6%), and colicin V (ColV; 60%). Twenty-two isolates (29.3%) were found to possess a combination of VAGs; iss, tsh, iroN, and iutA, in addition to other VAGs. To the best of our knowledge, this is the first report on the detection of virulence-associated genes and multidrug resistance among APEC isolates in Pakistan. In the future, the strains with the predominant set of VAGs can be used for colibacillosis diagnosis and as a potential vaccine candidate.

     

The effects of growth factors on proliferation of spermatogonial stem cells from Guangxi Bama mini‐pig

The objective of this study was to investigate the effects of different growth factors on the proliferation of Bama mini‐pig spermatogonial stem cells (SSCs) in vitro. The growth factors glial cell line‐derived neurotrophic factor (GDNF), leukaemia inhibitory factor (LIF), GDNF family receptor alpha‐1 (GFRα1) and basic fibroblast growth factor (bFGF) were investigated. The SSCs were seeded on SIM mouse embryo‐derived thioguanine‐ and ouabain‐resistant (STO) feeder layers. Cultivation of the cells were subjected to a factorial design of the growth factors GDNF + bFGF, GDNF + bFGF + GFRα1, LIF + bFGF and LIF + bFGF + GFRα1. The SSCs could propagate for 25 passages in the medium adding GDNF + bFGF + GFRα1, 22 passages in the medium adding GDNF + bFGF, 6 passages in the medium adding LIF + bFGF, or LIF + bFGF + GFRα1. qRT‐PCR analysis showed that the highest mRNA expression levels of NANOG, POU5F, DDX4, GFRα1 and UCHL1 were detected in the group adding GDNF + bFGF + GFRα1. The SSCs from the group adding GDNF + bFGF + GFRα1 also showed UCHL1‐, DBA‐ and CDH1‐positive staining. Moreover, Stra8 and Scp3 expression, and haploid peak were detected after induction of the SSCs from the group adding GDNF + bFGF + GFRα1. In conclusion, pig SSCs could be maintained for long term in the presence of GDNF, bFGF, and GFRα1.