Nutritional significance of amino acids,vitamins and minerals as nutraceuticals in poultry production and health – a comprehensive review

Nutraceuticals have gained immense importance in poultry science recently considering the nutritional and beneficial health effects of their constituents. Besides providing nutritional requirements to birds, nutraceuticals have beneficial pharmacological effects, for example, they help in establishing normal physiological health status, prevent diseases and thereby improve production performance. Nutraceuticals include amino acids, vitamins, minerals, enzymes, etc. which are important for preventing oxidative stress, regulating the immune response and maintaining normal physiological, biochemical and homeostatic mechanisms. Nutraceuticals help in supplying nutrients in balanced amounts for supporting the optimal growth performance in modern poultry flocks, and as a dietary supplement can reduce the use of antibiotics. The application of antibiotic growth enhancers in poultry leads to the propagation of antibiotic-resistant microbes and drug residues; therefore, they have been restricted in many countries. Thus, there is a demand for natural feed additives that lead to the same growth enhancement without affecting the health. Nutraceuticals substances have an essential role in the development of the animals’ normal physiological functions and in protecting them against infectious diseases. In this review, the uses of amino acids, vitamins and minerals as well as their mode of action in growth promotion and elevation of immune system are discussed.     

A logistic analysis of the factors determining the decision of smallholder farmers to intercrop: A case study involving rubber–tea intercropping in Sri Lanka

The factors influencing the decision of smallholder farmers to adopt new farming technologies were studied with reference to rubber–tea intercropping in Sri Lanka. Rubber–tea intercropping has been recommended previously to rubber farmers as a means to improve productivity and income during the early pre-tapping phase of rubber growth. Although crop trials have shown that the two crops are agronomically compatible and potentially produce a combined economic yield superior to the yield of a sole crop grown on the same area of land, there is little evidence of widespread adoption of this practice among smallholder farmers in Sri Lanka. The aim of the study was to determine the major factors that influence the decision to undertake rubber–tea intercropping and to construct a predictive model that describes the likelihood of adoption of intercropping by traditional smallholder rubber growers. A rapid rural appraisal (RRA) was undertaken based on semi-structured interviews of 90 smallholder farmers in the main rubber growing low wet zone of Sri Lanka. Among a number of factors shown to significantly influence the decision to intercrop tea with rubber, three were shown to operate independently, namely level of income, source of income (i.e. solely from own farm or from farm plus additional off-farm enterprises), and availability of land considered suitable for tea cultivation. A statistical model developed through correlation and logistic analysis, which predicts the likelihood of a smallholder adopting intercropping based on these factors, is presented and discussed. The most likely combination of circumstances (82% probability) under which rubber–tea intercropping is practiced is shown to be where the farmer’s income is greater than Rs. 10,000 per month, where the farmer’s income is based solely on own farm enterprises, and where more than 80% of the farmer’s land area was judged to be suitable for tea cultivation. Conversely, 30% of smallholder farmers that chose not to intercrop did possess land suitable for tea cultivation. Qualitative responses to the RRA indicated that limitation of technical knowledge was the main problem subsequently faced by rubber farmers who had adopted rubber–tea intercropping. Results indicate that there is need for both income support through farm subsidies and further agricultural extension services, if rubber–tea intercropping is to be adopted more widely in Sri Lanka. The wider usefulness of the developed logistic model in determining the likelihood of adoption of intercropping by smallholder farmers is discussed.     

Changes in Hormonal Balance: A Possible Mechanism of Pre‐Sowing Chilling‐Induced Salt Tolerance in Spring Wheat

There is a lack of knowledge about factors contributing to the chilling‐induced alleviatory effects on growth of plants under salt stress. Thus, the primary objective of the study was to determine whether chilling‐induced changes in endogenous hormones, ionic partitioning within shoots and roots and/or gaseous exchange characteristics is involved in salt tolerance of two genetically diverses of wheat crops. For this purpose, the seeds of two spring wheat (Triticum aestivum) cultivars, MH‐97 (salt intolerant) and Inqlab‐91 (salt tolerant) were chilled at 3°C for 2 weeks. The chilled, hydroprimed and non‐primed (control) seeds of the two wheat cultivars were sown in both Petri dishes in a growth room and in the field after treatment with 15 dS m^?1 NaCl salinity. Chilling was very effective in increasing germination rate and subsequent growth when compared with hydropriming and control under salt stress. Results from field experiments clearly indicated the efficacy of chilling over hydropriming in improving shoot dry biomass and grain yield in either cultivar, particularly under salt stress. This increase in growth and yield was related to increased net photosynthetic rate, greater potential to uptake and accumulate the beneficial mineral elements (K⁺ and Ca²⁺) in the roots and reduced uptake and accumulation of toxic mineral element (Na⁺) in the shoots of both wheat cultivars when grown under salt stress. Salt‐stressed plants of both wheat cultivars raised from chilled seed had greater concentrations of indoleacetic acid, abscisic acid, salicylic acid and spermine when compared with hydropriming and control. Therefore, induction of salt tolerance by pre‐sowing chilling treatment in wheat could be attributed to its beneficial effects on ionic homeostasis and hormonal balance. The results presented are also helpful to understand the chilling‐induced cross adaptation of plants in natural environments. Moreover, efficacy of pre‐sowing chilling treatment over hydropriming suggested its commercial utilization as a low risk priming treatment for better wheat crop production under stressful environments.     

Effects of simulated acid rain and root-knot nematode on tomato

Effects were examined of simulated acid rain and of root-knot nematode, Meloidogyne incognita race 1, on plant growth, yield, photosynthetic pigments and leaf epidermal characters of tomato ( Lycopersicon esculentum cv. Pusa Ruby). Sequential inoculation exposures (pre-, post-, and concomitant with nematode inoculation) were carried out in a greenhouse. Intermittent treatments of simulated acid rain at pH 3.2 caused white-to-tan irregular lesions on both the upper and lower surfaces of tomato leaves. The foliar symptoms were more pronounced on nematode-infected plants. Simulated acid rain (pH 3.2) and/or nematode infection suppressed plant growth, yield and pigment synthesis, the effects being greatest in post-inoculation treatments compared with simulated acid rain at pH 6.8. The total weight of fruits per plant was greatly suppressed owing to simulated acid rain or nematode infection. Chlorophyll a was found to be more sensitive to simulated acid rain or nematode infection than other leaf pigments. Root penetration, galling, egg mass production, and fecundity (number of eggs per egg mass) of M. incognita were enhanced at pH 5.6 and suppressed at pH 3.2 compared with pH 6.8. Nematode infection or simulated acid rain at pH 3.2 suppressed stomata and trichome development (number and size). Simulated acid rain treatments at pH 5.6 had a positive effect on number and size of trichomes, but a negative effect on stomata. The apertures of stomata were wider on tomato leaves exposed to simulated acid rain, especially at pH 3.2, than at pH 6.8.     

EHV-1 gene63 is not essential for in vivo replication in horses and mice, nor does it affect reactivation in the horse: short communication

The aim of this study was to investigate the role of immediate early gene (gene63) in the pathogenesis of equine herpesvirus 1 (EHV-1) acute and latent infections in equine and murine models. EHV-1 gene63 mutant virus (g63mut) along with EHV-1 (Ab4) was used for intracerebral and intranasal infection of 3 and 17-day-old mice. Both viruses were recovered at the same frequency from tissues after infection. Two Welsh ponies were infected via the intranasal route with each of the viruses. Acute infection was monitored by virus isolation from nasal swabs and peripheral blood leukocytes. Six weeks post infection, peripheral blood leukocytes were taken from ponies and in vitro reactivation was positive for both viruses. At autopsy, both viruses were isolated by co-cultivation from bronchial and submandibular lymph nodes. These findings indicate that the mutation of EHV-1 gene63 does not play a role in the establishment and reactivation from latency.     

Effects of sludge on germination and initial growth performance of Leucaena leucocephala seedlings in the nursery

A study was carried out to determine the influence of different types of sludges （municipal, industrial and residential） on field germination, growth and nodulation of L. leucocephala seedlings in the nursery. Before sowing of seeds, different combinations of sludges were incorporated with the nutrient deficient natural forest soils. Field germination, nodulation status and physical growth parameters of seedlings （shoot and root length, vigor index, collar diameter, leaf number, fresh and dry weight of shoot and root and total dry biomass increment） were recorded after three and six months of seed sowing. Field germination, nodulation status and growth parameters were varied significantly in the soil amended with sludges in comparison to control. The highest number of nodule was recorded from soil amended with residential sludge （ 1：1 ） and highest fresh and dry nodule weight was also found from the same combination in both three and six month old seedlings. In case of growth parameters, the highest growth was recorded from soil and residential sludge （ 1：1 ） combination compared to control. From the study, it can be recommended that soil amended with residential sludge （ 1：1 ） provide better field germination, growth and nodule formation of L. leucocephala in degraded soil.     

RNAi-mediated knockdown of INHBB increases apoptosis and inhibits steroidogenesis in mouse granulosa cells

Inhibins are members of the TGFβ superfamily and act as suppressors of follicle stimulating hormone (FSH) secretion from pituitary glands via a negative feedback mechanism to regulate folliculogenesis. In this study, the INHBB gene was knocked down by three RNAi-Ready pSIREN-RetroQ-ZsGreen vector- mediated recombinant plasmids to explore the effects of INHBB silencing on granulosa cell (GC) cell cycle, apoptosis and steroid production in vitro. Quantitative real-time polymerase chain reaction, Western blot, flow cytometry and ELISA were performed to evaluate the role of INHBB in the mouse GC cell cycle, apoptosis and steroid production in vitro. The results showed that the relative mRNA and protein expression of INHBB in mouse GCs can be significantly reduced by RNAi with pshRNA-B1, pshRNA-B2 and pshRNA-B3 plasmids, with pshRNA-B3 having the best knockdown efficiency. Downregulation of the expression of INHBB significantly arrests cells in the G1 phase of the cell cycle and increases the apoptosis rate in GCs. This was further confirmed by downregulation of the protein expressions of Cyclin D1, Cyclin E and Bcl2, while the protein expression of Bax was upregulated. In addition, specific downregulation of INHBB markedly decreased the concentration of estradiol and progesterone, which was further validated by the decrease in the mRNA levels of CYP19A1and CYP11A1. These findings suggest that inhibin βB is important in the regulation of apoptosis and cell cycle progression in granulosa cells. Furthermore, the inhibin βB subunit has a role in the regulation of steroid hormone biosynthesis. Evidence is accumulating to support the concept that inhibin βB is physiologically essential for early folliculogenesis in the mouse.     

梅PmARF17克隆及其在花发育中与内源激素的调控模式

【目的】分析梅PmARF17的生物学功能,探究梅花发育进程中其表达丰度与内源激素动态变化的关系,为梅花发育的调控研究提供依据。【方法】以梅品种‘大嵌蒂’为试材,克隆PmARF17,利用生物信息学软件分析基因结构、系统进化及其与其他物种同源蛋白的差异;亚细胞定位确定PmARF17蛋白在细胞中作用的部位;以梅品种‘大嵌蒂’和‘龙眼’不同发育阶段的花芽、叶芽、花器官为试材,利用qRT-PCR检测PmARF17时空表达模式,通过UPLC法测定IAA、GA₃、ABA、ZT含量的动态变化,并与PmARF17的表达进行相关性分析;克隆PmARF17启动子,分析启动子的顺式作用元件,利用瞬时表达解析PmARF17与GA₃的调控模式。【结果】从梅品种‘大嵌蒂’中克隆得到PmARF17,系统进化树分析表明PmARF17蛋白与其他植物的ARF蛋白序列高度同源;亚细胞定位表明其作用于细胞核和细胞膜上;qRT-PCR表达和内源激素含量的相关性分析表明,PmARF17的表达与IAA含量的变化趋势没有明显的相关性。PmARF17在雌蕊完好花芽中的表达水平相对不完全花芽显著上调,而GA₃含量与PmARF17的表达趋势一致。ABA和ZT含量总体上与PmARF17的表达呈相反的趋势,表明两者可能抑制PmARF17的表达。PmARF17启动子含有GA顺式元件,且具有启动活性和组织表达特异性,在花瓣、雄蕊及根部特异表达。【结论】 PmARF17可能是梅花发育的正调控基因,促进梅雌蕊的正常发育。PmARF17的表达可能受到GA₃的正调控,其可能通过作用于雄蕊和花瓣,进而影响梅的雌蕊发育进程。     

Expression profiling of transgenes (Cry1Ac and Cry2A) in cotton genotypes under different genetic backgrounds

Transgenic cotton carrying the Cry1Ac gene has revolutionized insect pest control since its adoption, although the development of resistance in insect pests has reduced its efficacy.  After 10 years of cultivating Bacillus thuringiensis (Bt) cotton with a single Cry1Ac gene, growers are on the verge of adopting Bt cotton that carries the double gene (Cry1Ac+Cry2A) due to its better effectiveness against insect pests.  Thus, the current study was designed to evaluate the role of each gene in the effectiveness of Bt cotton carrying the double gene.  The expression levels of the Cry1Ac and Cry2A genes were evaluated in the leaves of 10 genotypes (2 parents and 8 F₁ hybrids) at 30 days after sowing (DAS), while samples of leaves, bolls and flowers were taken from the upper and lower canopies at 70 and 110 DAS.  The F₁ hybrids were developed through reciprocal crosses between two Bt (CKC-1, CKC-2) and two non-Bt (MNH-786, FH-942) parents.  The differential expression of transgenes was evaluated through Enzyme Linked Immuno-Sorbent Assay (ELISA).  The results showed that the MNH786×CKC-1 hybrid had the highest concentrations of Cry1Ac gene at 30 DAS (3.08 µg g^–1) and 110 DAS (1.01 µg g^–1) in leaves.  In contrast, the CKC-2×MNH-786 hybrid showed the lowest concentrations of Cry1Ac gene at 30 DAS (2.30 µg g^–1) and 110 DAS (0.86 µg g^–1).  The F₁ hybrid FH-942×CKC-2 showed the highest concentrations of Cry2A gene at 30 DAS (8.39 µg g^–1) and 110 DAS (7.74 µg g^–1) in leaves, while the CKC-1×MNH-786 hybrid expressed the lowest concentrations of Cry2A gene at 30 DAS (7.10 µg g^–1) and 110 DAS (8.31 µg g^–1).  A comparison between the two stages of plant growth showed that leaves had the highest concentrations at 30 DAS, whereas the lowest concentrations were observed at 110 DAS for both genes in leaves.  When the expression pattern was compared between various plant parts in genotype CKC-2, it was found that leaves had higher concentrations of Cry1Ac (3.12 µg g^–1) and Cry2A (8.31 µg g^–1) at 70 DAS, followed by bolls (Cry1Ac (1.66 µg g^–1) and Cry2A (8.15 µg g^–1)) and flowers (Cry1Ac (1.07 µg g^–1) and Cry2A (7.99 µg g^–1)).  The genotype CKC-2 had higher concentrations of Cry1Ac (3.12 µg g^–1) and Cry2A (8.31 µg g^–1) in the upper canopy but less accumulation (2.66 µg g^–1 of Cry1Ac, 8.09 µg g^–1 of Cry2A) in the lower canopy at 70 DAS.  Similarly, at 110 DAS, the expression levels of Cry1Ac and Cry2A in upper and lower canopy leaves were 1.52 and 7.92 µg g^–1, and 0.99 and 7.54 µg g^–1, respectively.  Hence, the current study demonstrates that different genotypes showed variable expression for both of the Cry1Ac and Cry2A genes during plant growth due to different genetic backgrounds.  The Cry2A gene had three-fold higher expression than Cry1Ac with significant differences in expression in different plant parts.  The findings of this study will be helpful for breeding insect-resistant double-gene genotypes with better gene expression levels of Cry1Ac and Cry2A for sustainable cotton production worldwide.