Spiraea hypericifolia L. (Rosaceae) is an important plant having medicinal importance, and useful for growing on the forest-steppe due to soil strengthening property. However, its natural habitat has been affected due to human activities, and thus development of conservation strategies is needed, but for that correct identification of the species is a prerequisite. Thus, DNA barcoding using matK and rbcL was carried out for S. hypericifolia. The sequenced product size of matK and rbcL was 895 bp and 599 bp, respectively, and they were deposited to GenBank. For shoot regeneration, nodal explants were placed in Murashige and Skoog (MS) medium fortified with 0.5 mg L−1 6-benzyladenine (BA) for bud break. Shoot multiplication was carried out on MS and Linsmaier and Skoog (LS) media containing 0.2 or 0.5 mg L−1 BA or thidiazuron (TDZ), with or without 0.01 mg L−1 indole-3-butyric acid (IBA). Optimal shoot number of 15.00 ± 2.71 with 0.78 ± 0.09 cm shoot length and 6.62 ± 0.65 number of leaves were recorded in LS medium containing 0.5 mg L−1 BA. Rooting of shoots was accomplished on hormone-free ½MS medium, forming 2.57 ± 0.22 number of roots (90

The distribution and abundance of ectothermic mosquitoes are strongly affected by temperature, but mechanisms remain unexplored. We describe the effect of temperature on the transcriptome of Anopheles stephensi, an invasive vector of human malaria. Adult females were maintained across a range of mean temperatures (20 °C, 24 °C and 28 °C), with daily fluctuations of +5 °C and −4 °C at each mean temperature. Transcriptomes were described up to 19 days post-blood meal. Of the >3100 differentially expressed genes, we observed shared temporal expression profiles across all temperatures, suggesting their indispensability to mosquito life history. Tolerance to 20 and 28 ( + 5°C/−4°C) was associated with larger and more diverse transcriptomes compared to 24 ( + 5 °C/−4 °C). Finally, we identified two distinct trends in gene expression in response to blood meal ingestion, oxidative stress, and reproduction. Our work has implications for mosquitoes’ response to thermal variation, mosquito immune-physiology, mosquito-malaria interactions and the development of vector control tools. Effect of thermal variation and a blood meal on the size, composition and dynamics of the transcriptome of the ectotherm and major malaria vector, Anopheles stephensi

Prunus ulmifolia Franch. (Rosaceae) was investigated for its phytochemical composition from South-Eastern Kazakhstan for the first time. HPLC analysis confirmed rutin (0.88%) in ethanol extract, and the extract also exhibited antioxidant activity. The GC/MS analysis identified total 44 components from main groups e.g. oxygenated monoterpenes (51.06%), oxygenated sesquiterpenes (20.33%), non-terpene derivatives (18.71%), and sesquiterpene hydrocarbons (7.89%), and the maximum content was of acyclic alcoholic monoterpenoid citronellol (36.58%). The hierarchical cluster analysis (HCA) from previous reports and present study was used to demonstrate the variations between essential oil compositions in different Prunus species. It formed three main clusters, cluster I consisted of species with benzaldehyde as dominant component. Cluster II included plants with benzaldehyde as secondary component, and cluster III was of P. ulmifolia in which benzaldehyde was not detected. Further, the essential oil was assessed for cytotoxic and antimicrobial activities too, and it showed better cytotoxic but poor antimicrobial activities.

Abstract Background Like other oviparous organisms, the gonotrophic cycle of mosquitoes is not complete until they have selected a suitable habitat to oviposit. In addition to the evolutionary constraints associated with selective oviposition behavior, the physiological demands relative to an organism’s oviposition status also influence their nutrient requirement from the environment. Yet, studies that measure transmission potential (vectorial capacity or competence) of mosquito-borne parasites rarely consider whether the rates of parasite replication and development could be influenced by these constraints resulting from whether mosquitoes have completed their gonotrophic cycle. Methods Anopheles stephensi mosquitoes were infected with Plasmodium berghei, the rodent analog of human malaria, and maintained on 1% or 10% dextrose and either provided oviposition sites (‘oviposited’ herein) to complete their gonotrophic cycle or forced to retain eggs (‘non-oviposited’). Transmission potential in the four groups was measured up to 27 days post-infection as the rates of (i) sporozoite appearance in the salivary glands (‘extrinsic incubation period' or EIP), (ii) vector survival and (iii) sporozoite densities. Results In the two groups of oviposited mosquitoes, rates of sporozoite appearance and densities in the salivary glands were clearly dependent on sugar availability, with shorter EIP and higher sporozoite densities in mosquitoes fed 10% dextrose. In contrast, rates of appearance and densities in the salivary glands were independent of sugar concentrations in non-oviposited mosquitoes, although both measures were slightly lower than in oviposited mosquitoes fed 10% dextrose. Vector survival was higher in non-oviposited mosquitoes. Conclusions Costs to parasite fitness and vector survival were buffered against changes in nutritional availability from the environment in non-oviposited but not oviposited mosquitoes. Taken together, these results suggest vectorial capacity for malaria parasites may be dependent on nutrient availability and oviposition/gonotrophic status and, as such, argue for more careful consideration of this interaction when estimating transmission potential. More broadly, the complex patterns resulting from physiological (nutrition) and evolutionary (egg-retention) trade-offs described here, combined with the ubiquity of selective oviposition behavior, implies the fitness of vector-borne pathogens could be shaped by selection for these traits, with implications for disease transmission and management. For instance, while reducing availability of oviposition sites and environmental sources of nutrition are key components of integrated vector management strategies, their abundance and distribution are under strong selection pressure from the patterns associated with climate change. Graphical Abstract

Abstract Background Malaria, a disease caused by parasites of the genus Plasmodium, continues to impact many regions globally. The rise in resistance to artemisinin-based anti-malarial drugs highlights the need for new treatments. Ideally, new anti-malarials will kill the asymptomatic liver stages as well as the symptomatic blood stages. While blood stage screening assays are routine and efficient, liver stage screening assays are more complex and costly. To decrease the cost of liver stage screening, a previously reported luciferase detection protocol requiring only common laboratory reagents was adapted for testing against luciferase-expressing Plasmodium berghei liver stage parasites. Methods After optimizing cell lysis conditions, the concentration of reagents, and the density of host hepatocytes (HepG2), the protocol was validated with 28 legacy anti-malarials to show this simple protocol produces a stable signal useful for obtaining quality small molecule potency data similar to that obtained from a high content imaging endpoint. The protocol was then used to screen the Global Health Priority Box (GHPB) and confirm the potency of hits in dose–response assays. Selectivity was determined using a galactose-based, 72 h HepG2 assay to avoid missing mitochondrial-toxic compounds due to the Crabtree effect. Receiver-operator characteristic plots were used to retroactively characterize the screens’ predictive value. Results Optimal luciferase signal was achieved using a lower HepG2 seed density (5 × 103 cells/well of a 384-well microtitre plate) compared to many previously reported luciferase-based screens. While producing lower signal compared to a commercial alternative, this luciferase detection method was found much more stable, with a > 3 h half-life, and robust enough for producing dose–response plots with as few as 500 sporozoites/well. A screen of the GHPB resulted in 9 hits with selective activity against P. berghei liver schizonts, including MMV674132 which exhibited 30.2 nM potency. Retrospective analyses show excellent predictive value for both anti-malarial activity and cytotoxicity. Conclusions This method is suitable for high-throughput screening at a cost nearly 20-fold less than using commercial luciferase detection kits, thereby enabling larger liver stage anti-malarial screens and hit optimization make-test cycles. Further optimization of the hits detected using this protocol is ongoing.

Ingestion of an additional blood meal(s) by a hematophagic insect can accelerate development of several vector-borne parasites and pathogens. Most studies, however, offer blood from the same vertebrate host species as the original challenge (for e.g., human for primary and additional blood meals). Here, we show a second blood meal from bovine and canine hosts can also enhance sporozoite migration in Anopheles stephensi mosquitoes infected with the human- and rodent-restricted Plasmodium falciparum and P. berghei, respectively. The extrinsic incubation period (time to sporozoite appearance in salivary glands) showed more consistent reductions with blood from human and bovine donors than canine blood, although the latter's effect may be confounded by the toxicity, albeit non-specific, associated with the anticoagulant used to collect whole blood from donors. The complex patterns of enhancement highlight the limitations of a laboratory system but are nonetheless reminiscent of parasite host-specificity and mosquito adaptations, and the genetic predisposition of An. stephensi for bovine blood. We suggest that in natural settings, a blood meal from any vertebrate host could accentuate the risk of human infections by P. falciparum: targeting vectors that also feed on animals, via endectocides for instance, may reduce the number of malaria-infected mosquitoes and thus directly lower residual transmission. Since endectocides also benefit animal health, our results underscore the utility of the One Health framework, which postulates that human health and well-being is interconnected with that of animals. We posit this framework will be further validated if our observations also apply to other vector-borne diseases which together are responsible for some of the highest rates of morbidity and mortality in socio-economically disadvantaged populations.

Mushrooms are widely used in all possible fields, such as food, cosmetics, medicines and many more (Valverde et al., 2015; Zhang et al., 2016). Edible mushrooms are highly nutritive and can be consumed raw as well as boiled or fried (Wang et al., 1990). They are rich in fibres, vitamins and digestible proteins (Ouzouni et al., 2009; Reis et al., 2012). There are about 3,000 approximately edible mushrooms present on earth and among which approximately 200 species are consumed by humans (Kalač, 2013) and still need more to be highlighted and cultivated. Apart from being edible, mushrooms are widely used in pharmaceutical and nutraceutical industries as these are considered to be medicinal herbs. Globally mushrooms contribute approximately 25% of the molecules used by pharma industries to make medicines (Newman and Cragg, 2012). Mushrooms generally have high nutritious and medicinal value and have bioactive components, like 150–300 g of protein, 15–50 g of lipid and 75–150 g of dry weight of it (per kg). The nutritional energy is found to be least in their dry matter with only 300–500 Kcal per kg (Kalač, 2013).

Cotoneaster melanocarpus Fisch. ex A.Blytt (Rosaceae) is a deciduous shrub that is difficult to propagate through traditional methods. Thus, in the present study, in vitro regeneration utilizing nodal explants was attempted for the first time. Murashige and Skoog (MS) and Quoirin and Lepoivre (QL) medium supplemented with 0.2 or 0.5 mg L−1 6-benzyladenine (BA), 0.01 mg L−1 indole-3-butyric acid (IBA), and 0.5 mg L−1 gibberellic acid (GA3) were utilized for shoot culture establishment. Optimum 8.83 ± 0.66 shoots with 0.87 ± 0.10 cm length and 4.27 ± 0.37 leaves per explant were recorded after 40 d in MS medium fortified with 0.2 mg L−1 BA, 0.01 mg L−1 IBA, and 0.5 mg L−1 GA3. These shoots were then rooted in ½MS medium augmented with 0.5 mg L−1 IBA which formed 4.03 ± 0.29 roots per shoot (100

The agricultural practices of breeding, farm management and cultivation have improved production, to a great extent, in order to meet the food demands of a growing population. However, the newer challenges of climate change, global warming, and nutritional quality improvement will have to be addressed under a new scenario. Plant biotechnology has emerged as a reliable tool for enhancing crop yields by protecting plants against insect pests and metabolic engineering through the addition of new genes and, to some extent, nutritional quality improvement. Plant tissue culture techniques have provided ways for the accelerated clonal multiplication of selected varieties with the enhanced production of value-added plant products to increase modern agriculture. The in vitro propagation method has appeared as a pre-eminent approach for the escalated production of healthy plants in relatively shorter durations, also circumventing seasonal effects. However, there are various kinds of factors that directly or indirectly affect the efficiency of in vitro regeneration like the concentration and combination of growth regulators, variety/genotype of the mother plant, explant type, age of seedlings and other nutritional factors, and elicitors. Nanotechnology as one of the latest and most advanced approaches in the material sciences, and can be considered to be very promising for the improvement of crop production. Nanomaterials have various kinds of properties because of their small size, such as an enhanced contact surface area, increased reactivity, stability, chemical composition, etc., which can be employed in plant sciences to alter the potential and performance of plants to improve tissue culture practices. Implementing nanomaterials with in vitro production procedures has been demonstrated to increase the shoot multiplication potential, stress adaptation and yield of plant-based products. However, nanotoxicity and biosafety issues are limitations, but there is evidence that implies the promotion and further exploration of nanoparticles in agriculture production. The incorporation of properly designed nanoparticles with tissue culture programs in a controlled manner can be assumed as a new pathway for sustainable agriculture development. The present review enlists different studies in which treatment with various nanoparticles influenced the growth and biochemical responses of seed germination, as well as the in vitro morphogenesis of many crop species. In addition, many studies suggest that nanoparticles can be useful as elicitors for elevating levels of important secondary metabolites in in vitro cultures. Recent advancements in this field also depict the suitability of nanoparticles as a promising carrier for gene transfer, which show better efficiency than traditional Agrobacterium-mediated delivery. This review comprehensively highlights different in vitro studies that will aid in identifying research gaps and provide future directions for unexplored areas of research in important crop species.

Plant breeding through hybridization in cassava is facing a problem due to inconsistent flowering, and also the donor genes controlling superior traits are limited. An alternative method of breeding is through genetic transformation, and regeneration via somatic embryogenesis is promising route to achieve this. As somatic embryogenesis in cassava is genotype-specific, in the present study a protocol has been developed for UJ-3 and BW-1 genotypes. Immature sterile leaves from 7-10 days axillary shoots in a pre-condition medium were used as an explant. Leaves were inoculated on Murashige and Skoog (MS) medium containing picloram (0.0, 7.5, 10.0, 12.5, and 15.0 mg/L) and 1-naphthalene acetic acid (NAA, 6 mg/L) for induction of somatic embryos (SEs). Genotype BW-1 showed best results as early callus formation time i.e. 8.04±0.32 days after induction (dai) compared to UJ-3 (8.67±2.13 dai). The callus fresh weight (0.64 g) was also higher in BW-1 than UJ-3 (0.38 g) after 4 weeks in callus induction medium (CIM), and the callus formation ranges between 85.19±3.70 to 96.30±3.70% for both genotypes. Subculturing embryogenic callus to MS+CuSO4 (4 µM) + picloram (6 mg/L) +NAA (0.5 mg/L) (SK1 medium) germinated maximum SEs in BW-1 (46.56±36.86), whereas the number was less for UJ-3 (11.89±11.90). Further, shoots were developed from green cotyledons followed by hardening and acclimatization of plantlets.

Aflatunia ulmifolia (Franch.) Vassilcz. (Rosaceae) is an ornamental plant commonly used for landscaping and is known to form hybrids with other species that have characteristics like frost-resistant and non-rotting rootstocks. However, it is listed as an endangered species in Kazakhstan and is difficult to grow through traditional propagation; thus, in the present study, in vitro regeneration using seed and nodal explants was attempted. Murashige and Skoog (MS), Woody Plant Medium, and Gamborg medium supplemented with different plant growth regulators were utilized. Maximum germination (51.7±4.4

Solanum nigrum L. is an important medicinal plant of India and in the present study, leaves were used as an explant to establish shoot cultures. Murashige and Skoog’s18 (MS) medium supplemented with 20 μM kinetin (Kn) regenerated optimum 93.80 ± 0.89 shoots (100% response) within eight weeks through indirect organogenesis. Further, leaves were placed under simulated microgravity conditions and shoot regeneration was achieved within 16th days in both horizontal and vertical conditions. But horizontal position was better as compared to vertical position and it formed 11.60 ± 0.46 shoots. The in vitro shoots were successfully rooted in ½MS static medium fortified with sucrose (1%) and indole-3-butyric acid (IBA, 2 μM) which induced 39.40 ± 1.08 roots (100% response) within four weeks. This study can be used for mass-propagation of the species, extraction of valuable metabolites and to observe the effect of microgravity on plants.

Abstract Background Sporozoites isolated from the salivary glands of Plasmodium-infected mosquitoes are a prerequisite for several basic and pre-clinical applications. Although salivary glands are pooled to maximize sporozoite recovery, insufficient yields pose logistical and analytical hurdles; thus, predicting yields prior to isolation would be valuable. Preceding oocyst densities in the midgut is an obvious candidate. However, it is unclear whether current understanding of its relationship with sporozoite densities can be used to maximize yields, or whether it can capture the potential density-dependence in rates of sporozoite invasion of the salivary glands. Methods This study presents a retrospective analysis of Anopheles stephensi mosquitoes infected with two strains of the rodent-specific Plasmodium berghei. Mean oocyst densities were estimated in the midguts earlier in the infection (11–15 days post-blood meal), with sporozoites pooled from the salivary glands later in the infection (17–29 days). Generalized linear mixed effects models were used to determine if (1) mean oocyst densities can predict sporozoite yields from pooled salivary glands, (2) whether these densities can capture differences in rates of sporozoite invasion of salivary glands, and (3), if the interaction between oocyst densities and time could be leveraged to boost overall yields. Results The non-linear effect of mean oocyst densities confirmed the role of density-dependent constraints in limiting yields beyond certain oocyst densities. Irrespective of oocyst densities however, the continued invasion of salivary glands by the sporozoites boosted recoveries over time (17–29 days post-blood meal) for either parasite strain. Conclusions Sporozoite invasion of the salivary glands over time can be leveraged to maximize yields for P. berghei. In general, however, invasion of the salivary glands over time is a critical fitness determinant for all Plasmodium species (extrinsic incubation period, EIP). Thus, delaying sporozoite collection could, in principle, substantially reduce dissection effort for any parasite within the genus, with the results also alluding to the potential for changes in sporozoites densities over time to modify infectivity for the next host.

Abstract The aim of this research is to assess the current status, potential and problems of human resources for forestry in municipal state forest management institutions (MSFMIs) of Kazakhstan. A total of 120 MSFMIs are functional in Kazakhstan, with almost 5.5 thousand employees, of which 6.1% are women. Overall 63% are foresters, 13% are masters of the wood, 13% are engineers of forestry of all categories, 7% are forest wardens and 4% are directors. The quality of education of forest workers also varies and a minority of forest workers has a higher education. The scenario also suggested that almost 3000 specialists had taken up the advanced training courses from 2003 to 2019. According to a survey of employees of the MSFMIs of two pilot oblasts (Almaty and East Kazakhstan), the average age of an employee is 44.7 years and 79.2% of employees have a work experience of up to 15 years. The main issues and difficulties observed in the work of MSFMIs staff were low wages, lack of modern technique and equipment for forest service, lack of systematic continuing education of employees, poor quality of education, as well as staff turnover, especially foresters, which consequently affects their professional level.

Euonymus verrucosus Scop. (Celastraceae) is a boreal European-Mediterranean-Asia Minor forest relict species and is listed in the Red Data Books of Kazakhstan and some regions of the Russian Federation. Thus, in the present study, in vitro regeneration of E. verrucosus utilizing young shoot explants was attempted for the first time. Initially, the young shoots (0.8-1.0 cm length) were placed in woody plant medium (WPM) and Murashige and Skoog's (MS) medium fortified with different concentrations of cytokinins, such as 6-benzylaminopurine (BAP), kinetin (Kn) and thidiazuron (TDZ), to induce axillary buds. The optimum induction was 75.0 +/- 2.9% and the apical growth was 3.74 +/- 0.06 cm for young shoots after 4 weeks in WPM medium supplemented with BAP (1.0 mg L-1). Simultaneously, during the formation of axillary shoots, callus induction was observed at the nodal base. Subculturing this callus on fresh medium every 4 weeks generated total six adventitious shoots with 0.70 +/- 0.02 cm length after 12 weeks in the presence of BAP (1.0 mg L-1). When the adventitious shoots were isolated and transferred again to WPM medium containing 2.0 mg L-1 Kn, 2.95 +/- 0.16 shoots were formed with 51.2% regeneration frequency within 4 weeks. These microshoots were then rooted ex vitro and an optimum 73.3% rooting was achieved within 4 weeks in a substrate highbog peat with a 0.5 cm layer of vermiculite. This micropropagation protocol will be helpful for the conservation of the genetic resources and biodiversity for E. verrucosus.

Co-infected hosts, individuals that carry more than one infectious agent at any one time, have been suggested to facilitate pathogen transmission, including the emergence of supershedding events. However, how the host immune response mediates the interactions between co-infecting pathogens and how these affect the dynamics of shedding remains largely unclear. We used laboratory experiments and a modeling approach to examine temporal changes in the shedding of the respiratory bacterium Bordetella bronchiseptica in rabbits with one or two gastrointestinal helminth species. Experimental data showed that rabbits co-infected with one or both helminths shed significantly more B. bronchiseptica, by direct contact with an agar petri dish, than rabbits with bacteria alone. Co-infected hosts generated supershedding events of higher intensity and more frequently than hosts with no helminths. To explain this variation in shedding an infection-immune model was developed and fitted to rabbits of each group. Simulations suggested that differences in the magnitude and duration of shedding could be explained by the effect of the two helminths on the relative contribution of neutrophils and specific IgA and IgG to B. bronchiseptica neutralization in the respiratory tract. However, the interactions between infection and immune response at the scale of analysis that we used could not capture the rapid variation in the intensity of shedding of every rabbit. We suggest that fast and local changes at the level of respiratory tissue probably played a more important role. This study indicates that co-infected hosts are important source of variation in shedding, and provides a quantitative explanation into the role of helminths to the dynamics of respiratory bacterial infections.

In the flora of Kazakhstan there are many medicinal plants, of which the genus Artemisia (Asteraceae) includes 81 species. In the current study, chemical composition of essential oil from aerial parts of Artemisia austriaca Jacq. collected from different sites of Northern Kazakhstan was determined using GC-MS analysis. The chemical analysis demonstrated that the oil was dominated by oxygenated monoterpenes amounting to 39.49-59.20% with camphor (7.03-20.52%), 1,8-cineole (8.95-13.55%), α-thujone (3.16-25.78%) and β-thujone (0.87-9.92%) as major constituents. The results also suggested that there was difference in composition as well as amount among different sites depending on pH and organic matter of the soil. Further chemometric analysis using hierarchical cluster analysis (HCA) of A. austriaca essential oil compositions from the published literature as well as the composition from present study were used in order to demonstrate geographical variations in the composition of the essential oils. It showed the existence of two main clusters: mixture of α- and β-thujones (32.5±21.6%) / 1,8-cineole (13.9±7.8%) (Cluster I) and camphor (40.5±17.4%) / 1,8-cineole (19.4±9.5%) (Cluster II).

Harmonic convergence is a potential cue, female mosquitoes use to choose male mates. However, very little is known about the benefits this choice confers to offspring performance. Using Aedes aegypti (an important vector of human disease), we investigated whether offspring of converging parental pairs showed differences in immune competence compared to offspring derived from non-converging parental pairs. Here we show that harmonic convergence, along with several other interacting factors (sex, age, reproductive, and physiological status), significantly shaped offspring immune responses (melanization and response to a bacterial challenge). Harmonic convergence had a stronger effect on the immune response of male offspring than on female offspring. Further, female offspring from converging parental pairs disseminated dengue virus more quickly than offspring derived from non-converging parental pairs. Our results provide insight into a wide range of selective pressures shaping mosquito immune function and could have important implications for disease transmission and control.

A total of nine derivatives of 4-thiazolidinone were synthesized involving the reaction of benzene-1,2- diamine with 4-aminobenzoic acid followed by reaction with substituted benzaldehyde to get the Schiff bases. These synthesized Schiff bases were further reacted with thioglycolic acid to get the desired thiazolidinones (29-37). In addition to conventional synthesis, the microwave irradiation method has also been employedfor the synthesis of these compounds which provides not only pollution free and eco-friendly environment but also excellent yields. The results showed that 2-substituted thiazolidinone derivatives exhibit good antibacterial activity. It was also recorded that the compounds containing -Cl, -NO2 group with thiazolidinone nucleus are more active than other compounds of the synthesized series.

In the present study, nodes of Eclipta alba (L.) Hassk. were placed in Murashige and Skoog's (MS) medium fortified with different concentrations of plant growth regulators (PGRs). Amongst all the combinations tried, Murashige and Skoog 's (MS) medium fortified with N-6-benzyladenine (BA, 10 mu M) and indole-3-acetic acid (IAA, 1 mu M) formed maximum 18.40 +/- 0.84 shoots in 100% cultures. In vitro shoots derived from optimized medium were assessed for their biosynthetic potential using HPTLC fingerprinting which revealed that these shoots are also able to synthesize similar metabolites as in vivo shoots.Further in vitro shoots were rooted using different strengths of static and liquid MS media as well as in natural planting substrates of which 1/2MS liquid medium fortified with sucrose (1%) and indole-3-butyric acid (IBA, 8 mu M) induced optimum 40.50 +/- 1.38 roots within four weeks. The rooted shoots through in vitro and ex vitro rooting were successfully hardened and acclimatized in garden soil under greenhouse conditions.