Molecular characterisation of Theileria equi and risk factors associated with the occurrence of theileriosis in horses of Punjab (Pakistan)

Theileria equi (T. equi) is an obligate intra- and extra-erythrocytic parasite that causes equine theileriosis (ET) in equids. Equine theileriosis is considered a notifiable disease of global significance, a major constraint to the international movement of horses, and endemic in many countries. This disease may be difficult to diagnose, as it can produce variable and nonspecific clinical signs. A cross-sectional study was designed for the molecular characterisation of T. equi and to investigate the associated risk factors of ET accompanied by its consequences on haematological and sero-biochemical parameters. A convenience sampling of 500 blood samples were collected from ET suspect horses from January to December 2017. PCR was performed on all blood samples targeting the 18S rRNA gene of T. equi followed by sequencing; 9% animals tested positive with confirmed sequences. The isolates of this study showed high homology with Cuban, Russian and Brazilian isolates of T. equi (accession numbers KY111762.2 , MG551915.1 and KY952237.1 , respectively). Based on multivariate analysis, the principal risk factors consisted of absence of dogs on the premises and presence of tick infestation. The haemato-biochemical parameters showed a decrease in granulocytes and erythrocytes, and an increase in lymphocytes, monocytes, mean corpuscular volume, mean corpuscular haemoglobin, mean platelet volume, glucose, phosphorus and aspartate aminotransferase in positive horses. This is the first study which identified ET in Punjab (Pakistan) using molecular techniques and risk factors together with the haemato-biochemical variations in horses.     

Genetic Variation in Cadmium Accumulation and Tolerance among Wheat Cultivars at the Seedling Stage

Exploitation of genetic differences to select wheat cultivars’ pollution-safe from cadmium (Cd) contamination requires better understanding of Cd uptake and translocation patterns. For this purpose, 15 wheat cultivars were grown in nutrient solution and exposed to four levels of Cd, i.e., 0, 15, 30, and 45 µM. The plants were harvested after 2 weeks of Cd exposure. Root and shoot relative dry matter (DM) was decreased in most of the cultivars, but some cultivars did not exhibit any toxic symptoms. The lowest Cd concentration in shoots was recorded for Lasani-2008 and Iqbal-2000 while the highest for Sehar-2006 and Inqlab-91. Both root absorption and translocation accounted for regulating Cd concentration in shoots. There was no relationship between relative DM and Cd concentrations in roots and shoots. The results suggest that wheat cultivars with low shoot Cd concentration but higher tolerance, i.e. Lasani-2008 and Iqbal-2000, could be used in breading programs for low Cd wheat.     

Farmers' attitudes towards techniques for improving oestrus detection in dairy herds in South West England

Unidentified heats contribute to declining fertility rates in English dairy herds. Several techniques have been advocated to improve heat detection rates. Despite demonstrable technical efficacy and cost-effectiveness, uptake is low. A study in South West England used the Theory of Reasoned Action (TORA) to explore dairy farmers' attitudes and beliefs towards heat detection techniques. Few farmers were convinced that following prescribed observation times, milk progesterone testing and using pedometers would fit their system or improve on their current heat detection practices. Perceived difficulty of using a technique was not a constraint on adoption. Without promotion that addresses identified barriers and drivers to adoption, little change in current practice can be expected.     

Perspectives of folate biofortification of cereal grains

One sixth of the world’s population is suffering from hidden hunger that indicates a gross malnutrition particularly among children and women of third world countries. The deficiency of micro nutrients, especially iron (Fe) causes a number of ailments such as megaloblastic anemia and neural tube defects in poor population. There is a dire need to supplement iron in the diet. Current efforts implicate fortification of wheat flour and other grains with different iron formulations such as ethylenediaminetetraacetic acid (EDTA), FeSO₄ and elemental iron. However, all such interventions are not sustainable due to logistic and quality assurance problems in resource-limited settings. For a long term solution, development of crop plants with increased micronutrients and iron bioavailability is essential. Therefore, biofortification of cereal grains using translational genomics approaches for enhancement of folate through genome editing in cereals is inevitable to mitigate the folate deficiency in poor remote population in a cost effective manner.     

Mapping the QTLs underlying drought stress at developmental stage of sorghum (Sorghum bicolor (L.) Moench) by association analysis

This study was conducted to identify quantitative trait loci (QTLs) for drought tolerance in sorghum (Sorghum bicolor (L.) Moench) by association mapping using a simple sequence repeat (SSR)-marker-based diversity research set. Genotypic data for 98 SSR marker loci on ten chromosomes were used for the association analysis. The experiment was conducted under control (well-watered) and drought stress conditions, and the phenotypic values of 23 morphological traits were recorded. Drought tolerance was assessed by using a leaf drying score as a parameter of the tolerance/susceptibility: scores were assigned on a scale from 1 (most tolerant) to 9 (most susceptible). Under the control conditions, 17 QTLs associated with 12 traits were identified on chromosomes 1, 2, 4, 8, 9, and 10, with ?Log₁₀ (P) ranging from 2.5 to 7.6 and explaining 9.5–57.5 % of the total phenotypic variance for the traits. Under the drought stress conditions, nine QTLs associated with 8 traits were identified on chromosomes 1, 2, 3, and 10 that explained 9–61.2 % of the total phenotypic variance for the traits, with ?Log₁₀ (P) ranging from 2.5 to 3.5. QTLs for some traits were detected only under the drought stress condition, suggesting that these traits are important in drought tolerance. These QTLs could be used to further dissect the genetic and physiological basis of drought tolerance in sorghum.     

Thermal,swelling and stability kinetics of chitosan based semi-interpenetrating network hydrogels

The present study is focused on studying the swelling kinetics, thermal and aqueous stabilities, and determination of various forms of water in the chitosan (CS) and polyacrylonitrile (PAN) blend and semi-interpenetrating polymer network (sIPN). CS/PAN blend hydrogel films were prepared by solution casting technique. The blend film with optimum swelling properties was selected for the synthesis of sIPN. CS in the blend was crosslinked with the vapors of Glutaraldehyde (GTA) to prepare sIPN. The fabricated CS/PAN blend and sIPN hydrogels films were characterized with Fourier transform infrared (FTIR), thermal gravimetric analysis (TGA) and field emission scanning electron microscope (FESEM). The kinetics of swelling, bound and unbound waters and aqueous stability were determined experimentally. FESEM showed good miscibility between CS and PAN, FTIR showed no chemical interaction between CS and PAN; however, it did show a doublet for the sIPN, TGA showed improved thermal stability and swelling kinetic followed second order kinetics. The degree of swelling of the sIPN hydrogels samples at room temperature varied from ~2200 % (with a fair degree of stability (~30 %)) to ~1000 % (with high degree of aqueous stability (43 %)) with increase in the crosslinking time. The calculated unbound water (W_UB) max., for the blend was 52.3 % whereas for the bound (W_B) the max., was 41.9 %. However, for sIPN hydrogel films, the W_UB water decreased (max. 21.0 %) where as the W_B increased (max. 52.0 %). The decrease in W_UB and increase in the W_B is attributed to the formation of a compact structure and increase in the contact area between the water and polymers in sIPN hydrogels due to the induction of new water contacting point in these hydrogel films, respectively.     

Glycinebetaine Improves Chilling Tolerance in Hybrid Maize

Plant growth and development is hampered by various environmental stresses including chilling. We investigated the possibility of improving chilling tolerance in hybrid maize by glycinebetaine (GB) seed treatments. Maize hybrid (Hycorn 8288) seeds were soaked in 50, 100 and 150 mg l^?1 (p.p.m.) aerated solution of GB for 24 h and were dried back. Treated and untreated seeds were sown at 27 °C (optimal temperature) and at 15 °C (chilling stress) under controlled conditions. Germination and seedling growth was significantly hindered under chilling stress. Moreover, chilling stress significantly reduced the starch metabolism and relative water contents (RWC), and increased the membrane electrolyte leakage. However, activities of antioxidants (catalase, superoxide dismutase and ascorbate peroxidase) were increased under stress conditions. Seed treatments with GB improved the germination rate, root and shoot length, seedling fresh and dry weights, leaf and root scores, RWC, soluble sugars, α‐amylase activity and antioxidants significantly compared with untreated seeds under optimal and stress conditions. Induction of chilling tolerance was attributed to maintenance of high tissue water contents, reduced membrane electrolyte leakage, and higher antioxidant activities and carbohydrate metabolism. Seed treatment with 100 mg l^?1 GB was the best treatment for improving the performance of hybrid maize under normal and stress conditions compared with control and other levels used.     

Development of a monoclonal antibody-based co-agglutination test to detect enterotoxigenic Escherichia coli isolated from diarrheic neonatal calves

Escherichia coli (E. coli) strains were collected from young diarrheic calves in farms and field. Strains that expressed the K99 (F5) antigen were identified by agglutination tests using reference antibodies to K99 antigen and electron microscopy. The K99 antigen from a selected field strain (SAR-14) was heat-extracted and fractionated on a Sepharose CL-4B column. Further purification was carried out by sodium deoxycholate treatment and/or ion-exchange chromatography. Monoclonal antibodies to purified K99 antigen were produced by the hybridoma technique, and a specific clone, NEK99-5.6.12, was selected for propagation in tissue culture. The antibodies, thus obtained, were affinity-purified, characterized and coated onto Giemsa-stained Cowan-I strain of Staphylococcus aureus (S. aureus). The antibody-coated S. aureus were used in a co-agglutination test to detect K99+ E. coli isolated from feces of diarrheic calves. The specificity of the test was validated against reference monoclonal antibodies used in co-agglutination tests, as well as in ELISA. Specificity of the monoclonal antibodies was also tested against various Gram negative bacteria. The developed antibodies specifically detected purified K99 antigen in immunoblots, as well as K99+ E. coli in ELISA and co-agglutination tests. The co-agglutination test was specific and convenient for large-scale screening of K99+ E. coli isolates.     

Irrigation and Zn fertilizer management improves Zn phyto‐availability in various rice production systems

Zinc (Zn) is an important micronutrient for rice (Oryza sativa L.) production and its deficiency has been observed in various production systems. High grain Zn concentration is equally important for high rice yield and human health. In this work, the effects of Zn fertilization on seedling growth, grain yield, grain Zn concentration, and their association with root traits were studied under alternate wetting and drying (AWD), aerobic rice (AR), system of rice intensification (SRI), and continuous flooding (CF). Zinc fertilization (15 kg ha^?1) improved nursery seedlings chlorophyll and Zn concentrations, root length, and number of roots with highest values observed in CF. At harvesting, maximum plant height, panicle length, total and panicle bearing tillers, and kernel yield were found with Zn addition in AWD and CF rice systems. Mid season drainage provided at maximum tillering and Zn fertilization increased its concentration in leaves, culms, panicles, and grains under CF and AR at physiological maturity. Most of Zn applied was allocated into culms and panicles, nevertheless, a significant increase in grain Zn concentration was also observed in all production systems. Association of leaf Zn with grain Zn concentration was stronger than with culm and panicle Zn. The results indicate that Zn application after rice nursery transplanting is more important for grain Zn enrichment in all rice systems than for increase in grain yield in all systems except AWD where grain yield was also increased. More grain yield in CF and AWD as compared to SRI and AR can also be attributed to decreased spikelet sterility and to better Zn phyto‐availability in these rice systems at physiological maturity.     

果胶酶澄清和田红葡萄汁工艺研究

[目的]利用果胶酶对和田红葡萄汁进行了澄清处理,为和田红葡萄汁的工业化生产提供科学依据.[方法]采用单因素试验及正交试验,研究了果胶酶用量、酶解温度和酶解时间对葡萄汁透光率的影响.[结果]果胶酶澄清和田红葡萄汁的最佳工艺条件为:果胶酶用量0.4 g/L、酶解温度45℃、酶解时间150 min.酶解后葡萄汁透光率高达94;以上.[结论]果胶酶澄清和田红葡萄汁效果较理想.澄清汁中不含果胶物质.酶解后营养成分损失较小.