Suitable Host Research Articles

CULTURABLE ENDOSYMBIOTIC BACTERIA FROM THE INDIAN LAC INSECT, KERRIA LACCA (KERR)

The Indian lac insect, Kerria lacca (Kerr) (Coccoidea: Tachardiidae) is a commercially important phytosuccivorous and sessile scale insect. Lac insects are cultured on suitable host plants in India and some Southeast Asian countries to produce lac. The lac insect harbours a number of endosymbionts. Isolation of culturable microbial endosymbionts and their identification through 16S rRNA has revealed sex and host-related differences of microbial species. Bacillus boroniphilus, Enterobacter cloacae and Staphylococcus sp. were found only in the lac insects reared on the plant host Cajanus cajan, whereas Bacillus firmus, Lysinibacillus xylanilyticus, Bacillus horneckiae and Bacillus velezensis were recorded only from Flemingia macrophylla. B. firmus and L. xylanilyticus were female-specific and B. horneckiae and B. velezensis were male-specific with Flemingia macrophylla as host; E. cloacae was female-specific and Bacillus boroniphilus and Staphylococcus sp. were male specific with C. cajan. Biochemical characteristics of the isolates, their genetic relationship with their taxonomic kin and their probable role, based on the information available about these endosymbionts in other hosts, have been studied.

Reactions of Citrullus amarus and Cucumis metuliferus to Meloidogyne chitwoodi, Meloidogyne enterolobii and Meloidogyne luci

Meloidogyne chitwoodi, M. enterolobii, and M. luci are present in some EU countries, with restricted distributions, and plant resistance can be used to manage these nematodes. Two pot experiments were conducted under controlled conditions for 56 d to assess the host suitability of two potential rootstocks, Cucumis metuliferus BGV11135 and Citrullus amarus BGV5167, to one isolate of each nematode. The susceptible cucumber (Cucumis sativus) ‘Dasher II’, watermelon (Citrullus lanatus) ‘Sugar Baby’ and tomato (Solanum lycopersicum) ‘Coração-de-Boi’ were included for comparisons. A histopathological study using confocal-laser microscopy was also conducted 15 d after nematode inoculations. In the pot test, the rootstocks showed lower numbers of galls, egg masses, and eggs per plant than their susceptible ones. Reproduction indices of the rootstocks varied from immune to moderately resistant, depending on the isolate-rootstock combination. In the histopathological study, M. enterolobii and M. luci induced similar numbers of giant cells (GC) per feeding site in all germplasms. However, GC volumes and numbers of nuclei in rootstocks were lower than in the susceptible germplasms. GCs induced by M. chitwoodi were only detected in susceptible cucumber. These results emphasize the potential of C. metuliferus and C. amarus as effective, eco-friendly strategies for managing root-knot nematodes, and show the complex these host-pathogen interactions.

Developing paediatric skills and knowledge in pharmacy education: an exploration of a novel work-based learning experience

Abstract Introduction Pharmacy education in the United Kingdom must adapt to produce independent prescribing pharmacists ready to join an evolving healthcare system. Current placement practices exclude approximately 21% of the population due to a lack of specific recommendations surrounding paediatric experience and knowledge within the MPharm programme.[1] In turn, students’ current experiences may be limited by lack of interaction with unique learning outcomes offered by some healthcare settings, such as paediatric hospitals.[2] To promote student exposure in an overlooked speciality, novel approaches to work-based learning can be utilised. Aim This study aimed to explore pharmacy students’ experiences of work-based learning in a paediatric hospital setting. Methods In October 2022, fourth-year MPharm students at one school of pharmacy were invited to undertake work-based learning sessions across one academic year. The sessions aimed to develop students’ paediatric consultations skills and knowledge. Sessions consisted of a briefing, ward activities, scaffolded consultations with children and carers, and debriefs with a clinical supervisor. Debriefs included students reporting clinical information, required action and learning outcomes. All debriefs provided by students were transcribed by a clinical supervisor using a spreadsheet which recorded the date, ward visited, patient details, student handover, follow-up (if required) and learning outcomes. Data was initially cleaned, quality checked, and underwent content analysis to identify patterns and key themes to describe student experiences. Results Seventy-four students took part in sessions and delivered 233 consultations covering the medical history of the patient (76%, n=177), with varied levels of completeness. Students were exposed to acute conditions (41%, n=96) and chronic conditions (33%, n=76), with 13% (n=30) still awaiting diagnosis. Forty-eight percent (n=81) of learning points related to the pathology, diagnosis and symptoms of conditions, 24% (n=41) to medicines, 15% (n=25) to patient care, 11% (n=18) to non-clinical experiences and 2% (n=4) to other outcomes. In addition to carrying out ward activities, students underwent the processing of experiences during post-session debriefs: “It’s uncomfortable seeing a child struggle to breathe” [P131]. The process of active reflection was also evidenced in debriefs: “I felt very anxious, like a tightness in the chest, to hear that a child had a short life expectancy” [P145]; “I realised they had zero cultural competence after seeing a patient from the Middle East with jaundice” [P233]. Conclusion The study demonstrates a proof of concept that students can be exposed to complex care needs and challenging consultations under indirect supervision, demonstrating the paediatric setting to be a suitable work-based learning host. However, findings are limited to a single cohort of students at a single site, meaning transferability may be limited. Future studies could focus on longitudinal educational and emotional outcomes of students by measuring clinical knowledge, competence and confidence. Utilising post-session debriefs with peers and supervisors created a space to share both pharmaceutical and emotional learning points, aiding in managing the cognitive load of students. This experience not only highlights the requirement of paediatric exposure in pharmacy education programmes to aid the students’ future practice, but the importance of supervised reflective activities following work-based learning experiences.

Advanced K3V2(PO4)2O2F cathode for rechargeable potassium-ion batteries with high energy density

Potassium-ion batteries (PIBs) have emerged as promising candidates for cost-effective and sustainable energy-storage systems. Nevertheless, limited by the large K+ radius, PIBs have great difficulty in figuring out and designing suitable host materials. Herein, a suitable cathode material K3V2(PO4)2O2F (KVPOF) for PIBs has been carefully prepared. It exhibits a high specific capacity close to the theoretical value, 116.3 mAh/g at 20 mA/g within the voltage window of 2.0–4.5 V vs K+/K, corresponding to a de-/intercalation process of ∼2 mol K+ per formula unit. In addition, it presents an average operating voltage plateau of about 3.5 V, resulting in an energy density of about 410 Wh/kg. The crystal structure and phase transition are revealed by in situ x-ray diffraction, and the structure is found to be fully reversible during the de-/intercalation of K+. Furthermore, the potential of KVPOF cathode for applications at low temperatures was explored, and the full cell matched with graphite anode demonstrated fair electrochemical performance. The experimental results suggest the feasibility of using KVPOF as cathode material for rechargeable PIBs.

The importance of host weed species for root-knot nematodes, Meloidogyne spp. Göldi, 1897 (Tylenchida: Heteroderidae) in banana plantations

Banana is a significant economic source in Türkiye. Root-knot nematodes, Meloidogyne spp. Göldi, 1897 (Tylenchida: Heteroderidae) are the important pests in banana fields. This study was conducted from 2021 to 2022 to elucidate the relationship between root-knot nematodes and weed species in bananas in the Mediterranean Region. 2% of the banana production area in Adana, Antalya, Hatay and Mersin were surveyed regularly for this purpose. 1m2 frames were placed within a 2m radius around banana plants in the sampled areas. The host of root-knot nematodes with weed species identified within the frames was examined. Survey results indicated that Amaranthus retroflexus L. (46.34%), Portulaca oleracea L. (40.63%), and Solanum nigrum L. (37.84%) were the weed species most infected with root-knot nematodes. Furthermore, molecular analyses revealed that Abutilon theophrasti Medik., Amaranthus spp., Cucumis melo var. agrestis Naudin., Erodium cicutarium (L.) L'Hér. ex Aiton, Kickxia commutata (Bernh. ex Rchb.) Fritsch, Malva spp., Mercurialis annua L., P. oleracea, S. nigrum, and Sonchus oleraceus L. were suitable hosts for root-knot nematodes. This study is an important step in understanding the interaction between root-knot nematodes and weeds in banana. The presence of weed species in agricultural fields should be considered as they may support nematode populations and pose a threat to subsequent crops. Therefore, the implementation of weed control strategies could help producers to control nematode populations.

The effect of potassium on aluminous phase stability in the lower mantle

The aluminous calcium-ferrite type phase (CF) and new aluminous phase (NAL) are thought to hold the excess alumina produced by the decomposition of garnet in MORB compositions in the lower mantle. The respective stabilities of CF and NAL in the nepheline-spinel binary (NaAlSiO4\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$_{4}$$\\end{document}–MgAl2\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$_{2}$$\\end{document}O4\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$_{4}$$\\end{document}) are well established. However with the addition of further components the phase relations at lower mantle conditions remain unclear. Here we investigate a range of compositions around the nepheline apex of the nepheline-kalsilite-spinel compositional join (NaAlSiO4\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$_{4}$$\\end{document}–KAlSiO4\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$_{4}$$\\end{document}–MgAl2\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$_{2}$$\\end{document}O4\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$_{4}$$\\end{document}) at 28–78 GPa and 2000 K. Our experiments indicate that even small amounts of a kalsilite (KAlSiO4\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$_{4}$$\\end{document}) component dramatically impact phase relations. We find NAL to be stable up to at least 71 GPa in potassium-bearing compositions. This demonstrates the stabilizing effect of potassium on NAL, because NAL is not observed at pressures above 48 GPa on the nepheline-spinel binary. We also observe a broadening of the CF stability field to incorporate larger amounts of potassium with increasing pressure. For pressures below 50 GPa only minor amounts (<0.011(1)KK+Na+Mg\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$<0.011(1)\\frac{K}{K+Na+Mg}$$\\end{document}) of potassium are soluble in CF, whereas at 68 GPa, we find a solubility in CF of at least 0.088(3)KK+Na+Mg\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$0.088(3)\\frac{K}{K+Na+Mg}$$\\end{document}. This indicates that CF and NAL are suitable hosts of the alkali content of MORB compositions at lower mantle conditions. For sedimentary compositions at lower mantle pressures, we expect K-Hollandite to be stable in addition to CF and NAL for pressures of 28–48 GPa, based on our simplified compositions.

Host suitability for germination differs from that of later stages of development in a rare epiphytic orchid

We explore phorophyte suitability for germination and establishment of the epiphytic orchid, Psychilis kraenzlinii. We found that the orchid grows on a subset of the available tree species and shows preference for the endemic Machaonia portoricensis (Rubiaceae). The orchid preferred trees with smoother bark with high water holding capacity and low water retention capacity. Microclimatic conditions under which embryos began pre-germination stages mirrored that of the adult orchid, but germination did not, suggesting that suitable germination sites are not necessarily the best sites for later stages of development.

Host Plant Modulated Physio-Biochemical Process Enhances Adaptive Response of Sandalwood (Santalum album L.) under Salinity Stress.

Salinity is one of the most significant abiotic stress that affects the growth and development of high-value tree species, including sandalwood, which can also be managed effectively on saline soils with the help of suitable host species. Therefore, the current investigation was conducted to understand the physiological processes and antioxidant mechanisms in sandalwood along the different salinity gradients to explore the host species that could support sandalwood growth in salt-affected agro-ecosystems. Sandalwood seedlings were grown with ten diverse host species with saline water irrigation gradients (ECiw~3, 6, and 9 dS m-1) and control (ECiw~0.82 dS m-1). Experimental findings indicate a decline in the chlorophyll content (13-33%), relative water content (3-23%), photosynthetic (27-61%) and transpiration rate (23-66%), water and osmotic potential (up to 137%), and ion dynamics (up to 61%) with increasing salinity levels. Conversely, the carotenoid content (23-43%), antioxidant activity (up to 285%), and membrane injury (82-205%) were enhanced with increasing salinity stress. Specifically, among the hosts, Dalbergia sissoo and Melia dubia showed a minimum reduction in chlorophyll content, relative water content, and plant water relation and gas exchange parameters of sandalwood plants. Surprisingly, most of the host tree species maintained K+/Na+ of sandalwood up to moderate water salinity of ECiw~6 dS m-1; however, a further increase in water salinity decreased the K+/Na+ ratio of sandalwood by many-fold. Salinity stress also enhanced the antioxidative enzyme activity, although the maximum increase was noted with host plants M. dubia, followed by D. sissoo and Azadirachta indica. Overall, the investigation concluded that sandalwood with the host D. sissoo can be successfully grown in nurseries using saline irrigation water and, with the host M. dubia, it can be grown using good quality irrigation water.

Differential phase of Platypus quercivorus attacks in urban green space: A case study on 2 urban parks in Tokyo

The ambrosia beetle Platypus quercivorus (Maruyama) transmits the pathogenic fungus Raffaelea quercivora Kubono et Shin. Ito, causing Japanese oak wilt and subsequent tree dieback. In recent years, there has been a marked increase in mortality of secondary forests due to Japanese oak wilt, especially in suburban and urban parks, as P. quercivorus finds suitable host trees and environments. To prevent extensive mortality from P. quercivorus attacks, it is crucial to establish effective management strategies for secondary forests. In this study, we aimed to investigate the occurrence patterns of P. quercivorus attacks and identify effective management strategies in urban parks that have experienced recent outbreaks of Japanese oak wilt to find any hints for establishing effective management strategies. Over a two-year period, we surveyed the occurrence of P. quercivorus attacks on Quercus serrata Murray trees along walking trails and assessed surrounding conditions. Through data analysis, we identified significant factors influencing attacks. The results revealed distinct differences in host tree condition and attack severity between parks. Although the proportion of attacked trees in one park markedly increased over the two-year period, the other park remained stable, suggesting different attack phases between parks. In the former park, the occurrence of attacks was positively correlated with tree size, severity of nearby attacks, and nearby tree dieback but negatively correlated with surrounding tree density and occurrences of last year attack. Conversely, in the latter park, only occurrences of last year attack showed a significant negative correlation with attack occurrence. This discrepancy may be attributed to variations in the attack phase of P. quercivorus in each park. From a management perspective, park managers should establish flexible forest management strategies in suburban and urban parks to adapt to different attack phases.

Efficient Secretory Expression for Mammalian Hemoglobins in Pichia pastoris

Mammalian hemoglobins (HB) are a kind of heme-binding proteins that play crucial physiological roles in various organisms. The traditional techniques employed for the extraction of HB are expensive and time-consuming, while the yields of mammalian HB in previous reports were quite low. The industrial Pichia pastoris is a highly effective platform for the secretory expression of heterologous proteins. To achieve efficient secretory expression of HB in P. pastoris, multiple strategies were applied, including the selection of a suitable host, the screening of optimal endogenous signal peptides, the knockout of VPS10, VTH1, and PEP5, and the co-expression of Alpha-Hemoglobin Stabilizing Protein (AHSP). In addition, the conditions for producing HB were optimized at shaking-flask level (BMMY medium with 100 mg/L of hemin, 2% methanol, and 24 °C). Based on these conditions, the higher titers of bovine hemoglobin (bHB, 376.9 ± 13.3 mg/L), porcine hemoglobin (pHB, 119.2 ± 7.3 mg/L), and human hemoglobin (hHB, 101.1 ± 6.7 mg/L) were achieved at fermenter level. The engineered P. pastoris strain and comprehensive strategies can also be applied to facilitate the synthesis of other high-value-added hemoproteins or hemoenzymes.

Phorophyte preference of an epiphytic orchid (Dendrobium wangliangii) is independent of orchid mycorrhizal fungi that promote seed germination

Abstract Epiphytism is an example of an obligate interspecific interaction between plants, where a plant species grows on another plant species for physical support. In epiphytic orchids that rely on suitable host tree species (phorophytes) and mycorrhizal fungi for germination and growth, a distinct preference has been observed for particular host tree species. However, the factors driving this host preference remain unclear. In this study, we investigated whether mycorrhizal fungi that promote seedling development determine the preference of phorophytes in the epiphytic orchid Dendrobium wangliangii. First, we identified the preferred host tree species for this orchid. Next, in vitro and in situ germination experiments were conducted to identify potential fungi that promote seed germination. Finally, high‐throughput sequencing was used to assess the distribution of these fungi on both colonized host trees and potential host trees, including individual trees of the host species not yet colonized by the orchid and trees of another species growing in the vicinity of a colonized tree. Dendrobium wangliangii exhibited a preference for trees within the Fagaceae family. Two Fusarium species from the bark of host trees were identified as the primary fungi that promote seed germination. Successful protocorm formation and seedling development were observed exclusively on preferred phorophytes. Fungal community analyses further indicated that both Fusarium spp. occurred on all potential host trees, suggesting that the host preference of this epiphytic orchid is not constrained by the distribution of suitable mycorrhizal fungi. Synthesis: Regarding the epiphytes that depend on mycorrhizal fungi for successful seedling recruitment, we conclude that strong epiphyte–phorophyte associations do not arise from the limited distribution of mycorrhizal fungi but are more likely the result of other factors inherent in the phorophyte environment.

Biosynthesis of fragrance 2-phenylethanol from sugars by Pseudomonasputida

BackgroundPetrochemicals contribute to environmental issues, with concerns ranging from energy consumption and carbon emission to pollution. In contrast, microbial biorefineries offer eco-friendly alternatives. The solvent-tolerant Pseudomonasputida DOT-T1E serves as a suitable host for producing aromatic compounds, specifically l-phenylalanine and its derivative, 2-phenylethanol (2-PE), which find widespread applications in various industries.ResultsThis study focuses on enhancing 2-PE production in two l-phenylalanine overproducing strains of DOT-T1E, namely CM12-5 and CM12-5Δgcd (xylABE), which grow with glucose and glucose-xylose, respectively. To synthesize 2-PE from l-phenylalanine, these strains were transformed with plasmid pPE-1, bearing the Ehrlich pathway genes, and it was found higher 2-PE production with glucose (about 50–60 ppm) than with xylose (< 3 ppm). To understand the limiting factors, we tested the addition of phenylalanine and intermediates from the Ehrlich and shikimate pathways. The results identified intracellular l-phenylalanine as a key limiting factor for 2-PE production. To overcame this limitation, a chorismate mutase/prephenate dehydratase variant—insentive to feedback inhibition by aromatic amino acids—was introduced in the producing strains. This led to increased l-phenylalanine production and subsequently produced more 2-PE (100 ppm). Random mutagenesis of the strains also produced strains with higher l-phenylalanine titers and increased 2-PE production (up to 120 ppm). The improvements resulted from preventing dead-end product accumulation from shikimate and limiting the catabolism of potential pathway intermediates in the Ehrlich pathway. The study explored agricultural waste substrates, such as corn stover, sugarcane straw and corn-syrup as potential C sources. The best results were obtained using 2G substrates at 3% (between 82 and 100 ppm 2-PE), with glucose being the preferred sugar for 2-PE production among the monomeric sugars in these substrates.ConclusionsThe findings of this study offer strategies to enhance phenylalanine production, a key substrate for the synthesis of aromatic compounds. The ability of P.putida DOT-T1E to thrive with various C-sources and its tolerance to substrates, products, and potential toxicants in industrial wastes, are highlighted. The study identified and overcome possible bottlenecks for 2-PE production. Ultimately, the strains have potential to become efficient microbial platforms for synthesizing 2-PE from agro-industrial waste materials.

Effects of host plants on spotted lanternfly (Hemiptera: Fulgoridae) nymphal survival and development.

Spotted lanternfly, Lycorma delicatula (White), is an invasive planthopper from China, which was first detected in Berks County, PA, in 2014 and has since spread to adjacent states including New Jersey in 2018. Lycorma delicatula is a polyphagous species that gregariously feeds on over 172 known hosts. We investigated development on key host plants Ailanthus altissima (Miller) (Sapindales: Simaroubaceae), Juglans nigra (L.) (Fagales: Juglandaceae), Acer rubrum (L.) (Sapindales: Sapindaceae), and Vitis vinifera (L.) (Vitales: Vitaceae) for each instar to elucidate potential host use throughout the season and target monitoring efforts. Our study indicated significant differences in survivorship and time spent in each nymphal life stage between host plants. We applied a host suitability index as a function of survivorship and development for each host plant and instar, which indicated A. altissima and J. nigra as suitable hosts for all 4 nymphal instars. Vitis vinifera was highly suitable for first and second instars but had low indices for the third and fourth instars, although suitability of V. vinifera may have varied based on variety and age of the vine. Lycorma delicatula nymphs had the lowest survivorship and longest development time on A. rubrum across all 4 life stages, despite this being a preferred oviposition host. Host had a significant effect on the morphometrics we measured within the first and second instars.

Increased Expression Level of Human Blood Clotting Factor VIII Using NS0 Cell Line as a Host Cells.

Coagulation factor VIII (FVIII) is applied for spontaneous hemorrhaging inhibition and excessive bleeding after trauma in patients with hemophilia A. High-quality human recombinant factor VIII (rFVIII) has been produced relatively in large quantities in cultured mammalian cells. NS0 is one of the most common mammalian cell lines for therapeutic protein production. Production of rFVIII has increased due to low FVIII expression levels and rising demand for hemophilia A prophylactic treatment. Several methods have been developed to prevent cell cycle progression in mammalian cells for increased recombinant protein yields. The aim of the study was to investigate the level of recombinant BDD-FVIII expression in NS0 mouse myeloma cells. Additionally, the study aimed to determine the effects of chemical drugs, Mitomycin C, Lovastatin, and Metformin on the secretion of FVIII through cell cycle arrest. We cultured NS0 cells and transfected them with the 2 μg pcDNA3-hBDDFVIII plasmid by Lipofectamine 3000. The cells were treated with 10 μg.mL-1 Mitomycin C, 20 μM Lovastatin, and 20 mM Metformin separately. After 24 and 48 hours, the samples were collected and, protein expression was analyzed using RT-PCR, Dot blot, and ELISA. A higher protein expression level was observed in treated cells 24h and 48h after treatment with all three drugs. According to real-time PCR, Metformin treatment resulted in the highest expression level within 24 h (P=0.0026), followed by Mitomycin C treatment within 48 h (P=0.0030). The NS0 cell line can be regarded as a suitable host for FVIII production. FVIII protein expression level was increased by using Lovastatin, Metformin, and Mitomycin C drugs. Further investigations are suggested, and the potential application of these drugs to increase recombinant protein yield can be used to produce therapeutic proteins in the industry.

Risk assessment of Retithrips syriacus for the EU.

Following a request from the European Commission, the EFSA Panel on Plant Health performed a quantitative risk assessment for the EU of Retithrips syriacus (Mayet) (Thysanoptera: Thripidae), a polyphagous thrips, regarded as a tropical/subtropical pest occurring in several countries of Africa, South America, Asia and in the EU in Cyprus. The current risk assessment focused on potential pathways for entry, the climatic conditions allowing establishment, the expected spread capacity and the impact considering a time horizon of 10 years (2023-2032). The Panel identified the import of cut roses, persimmons, table grapes, as well as plants for planting of the genera Acalypha and Terminalia from third countries and those of Persea americana (avocado) from Israel as the most relevant entry pathways to consider. Over the next 10 years, an annual median estimate of 95 (90% Certainty Range, CR, ranging from 13 to 1832) potential R. syriacus founder populations per year are expected to successfully transfer to a suitable host in the EU NUTS2 regions where the climatic conditions are predicted as suitable for establishment; this value drops to a median of 4.6 founder populations per year (90% CR: 1 every 1.9 years - 85.6 per year) after considering the actual probability of establishment of a potential founder population. The estimated number of founder population per year is mostly driven by the import of cut roses and plants for planting. If such founder populations were to establish, R. syriacus is estimated to spread at a median rate of 0.05 km/year (90% CR 0.02-2.30 km/year) after a median lag phase of 1.1 years (90% CR 0.3-3.3 years). The overall impact on yield (expressed as % of the total agricultural production) directly attributable to R. syriacus when considering: (i) the main R. syriacus hosts in the EU, (ii) the areas of the EU where establishment is possible, (iii) the current agricultural practices and (iv) the evidence of impact from the countries where the pest is established for a long time, was estimated at 0.065% as the median value of the uncertainty distribution (90% CR 0.001%-0.571%). Options for risk reduction are discussed, but the effectiveness was not quantified.

First report of Root-Knot Nematode Meloidogyne enterolobii infecting coriander in Shaanxi, Northern China.

Coriander (Coriandrum sativum), which can be used for its root, stem, and leaf as both food and medicine (Prachayasittikul et al. 2018), is widely cultivated in China. The coriander cultivation area of Guanzhong region, including Xi' an, Xianyang, and Weinan, is 20 million m2, which accounts for 85.7% of the total cultivation area in Shaanxi. In September 2022, obvious galls were observed on the roots of coriander plants (cv. Xiaoye) growing in a field in Huyi District, Xi' an City (34°1'26.4"N, 108°31'58.8"E). The diseased plants did not show obvious above-ground symptoms. To identify the species, second-stage juveniles (J2s) and males were collected from soil in the root zone, and adult females were isolated from galls of diseased roots. The perineal patterns of adult females (n = 20) were round to oval, with high dorsal arches and no obvious lateral lines were observed. Morphological measurements of females (n = 20) included body length (L) = 682 ± 56 (554 to 780) μm, body width (BW) = 522 ± 45 (420 to 597) μm, stylet = 14.9 ± 0.9 (13.4 to 16.3) μm, dorsal pharyngeal gland orifice to stylet base (DGO) = 5.3 ± 0.5 (4.3 to 6.3) μm, vulval slit length = 26 ± 2.8 (20 to 32) μm, vulval slit to anus distance = 21 ± 1.7 (18.5 to 26) μm. Measurements of males (n = 8) were L = 1398 ± 57 (1308 to 1450) μm, BW = 28 ± 2.9 (23 to 32) μm, stylet = 16.1 ± 0.8 (15.3 to 17.3) μm, DGO = 4.5 ± 0.5 (3.5 to 4.9) μm, spicules = 27 ± 1.1 (26 to 29) μm. Measurements of J2s (n = 20) were as follows: L = 434 ± 16.8 (391 to 477) μm, BW = 15.6 ± 0.9 (13.7 to 17.3) μm, stylet = 12.6 ± 0.6 (11.3 to 13.6) μm, DGO = 3.9 ± 0.3 (3.4 to 4.5) μm, tail = 52 ± 4.0 (47 to 60) μm, hyaline tail length = 15.6 ± 1.3 (13.6 to 18.6) μm. These morphological characteristics were consistent with those described for Meloidogyne enterolobii (Yang and Eisenback 1983). Ten females were put in 10 tubes for DNA extraction following Htay et al. (2016). The ITS-rDNA sequence was amplified using the primers 18S/26S (Vrain et al. 1992). A 765 bp fragment was obtained and the sequence (GenBank OR789453) was 99.87% identical to sequences of M. enterolobii (MT406251 and MT067559). The mtDNA CoxII-16S sequence was amplified using primers C2F3/1108 (Powers and Harris, 1993). The sequence was 705 bp (OR795028) and 100% identical to sequences of M. enterolobii (MK455870 and MZ643270). A single 236 bp fragment was amplified using species-specific primers Me-F/Me-R, confirming the species as M. enterolobii (Long et al. 2006). The infection test was conducted in a greenhouse at 27 ± 2℃. Eight 2-week-old coriander plants (cv. Xiaoye) were individually grown in pots filled with sterilizer soil and inoculated with 800 J2s hatched from collected M. enterolobii egg masses. Forty-five days after nematode inoculation, the inoculated plants had galled roots like those observed in the field. The reproduction factor (final population density/initial population density) was 11.9 ± 2.0, indicating coriander was a suitable host for M. enterolobii. No symptoms were observed in controls. To our knowledge, this is the first known natural infection of coriander with M. enterolobii in China. M. enterolobii has been reported on various crops in southern provinces of China (EPPO, 2023). Considering the high level of agricultural trade between different regions, there is a high risk of M. enterolobii transmission to Guanzhong region through infested soil and susceptible plant materials. Further monitoring and research on effective control strategies are needed to prevent the spread of this nematode.

Expression and functional characterization of chimeric recombinant bovine follicle-stimulating hormone produced in Leishmania tarentolae.

Assisted reproductive techniques are routinely used in livestock species to increase and enhance productivity. Ovarian hyperstimulation is a process that currently relies on administering pituitary-derived follicle-stimulating hormone (FSH) or equine chorionic gonadotropin in combination with other hormones to promote the maturation of multiple follicles and thereby achieve superovulation. The use of partially purified preparations of FSH extracted from natural sources is associated with suboptimal and variable results. Recombinant FSH (rFSH) has been produced in a variety of heterologous organisms. However, attaining a bioactive rFSH of high quality and at low cost for use in livestock remains challenging. Here we report the production and characterization of a single chain bovine rFSH consisting of the β- and α-subunit fused by a polypeptide linker (scbFSH) using Leishmania tarentolae as heterologous expression system. This unicellular eukaryote is non-pathogenic to mammals, can be grown in bioreactors using simple and inexpensive semisynthetic media at 26°C and does not require CO2 or bovine serum supplementation. Stable cell lines expressing scbFSH in an inducible fashion were generated and characterized for their productivity. Different culture conditions and purification procedures were evaluated, and the recombinant product was biochemically and biologically characterized, including bioassays in an animal model. The results demonstrate that L. tarentolae is a suitable host for producing a homogeneous, glycosylated and biologically active form of scbFSH with a reasonable yield.

Genetic and pathogenic characterization of Rhizoctonia solani AG-4 isolates obtained from common bean

Common bean (Phaseolus vulgaris L.) is the most important grain legume. Crop production of common bean is affected by a number of diseases, such as Rhizoctonia root rot, being detected worldwide. Herein, we aimed to genetically and pathologically characterize Rhizoctonia solani isolates obtained from different provinces in Turkey. Anastomosis groups (AG) and subgroups of isolates were identified based on the sequence analysis of the rDNA-ITS region. The most prevalent subgroup was AG-4 HGIII, according to BLAST analysis and the phylogeny of resultant sequences, followed by AG-4 HGI and AG-4 HGII, respectively. iPBS retrotransposons highly supported the phylogenetic tree and provided a high level of genetic variability among isolates to discriminate AG subgroups. The results indicated that the iPBS DNA marker system based on retrotransposons could be used to discriminate Rhizoctonia isolates relying on AG grouping. The virulence of the pathogen isolates changed from 3.67 to 5 based on the agar-plate assay on the susceptible cv. Gina. The reactions of thirty common bean cultivars were also evaluated against the most aggressive isolate in each subgroup. The reaction assay showed significant differences among both isolates and cultivars; the highest disease severity among cultivars was observed to AG-4 HGI, followed by AG-4 HGIII and AG-4 HGII, respectively. None of the cultivars showed resistance to all AG subgroups. Screening of AGs among the isolates and selecting cultivars are critical to managing Rhizoctonia disease due to the range of host suitability according to AG subgroups.

Hidden diversity of cestodes in wild African carnivores: I. Non-taeniid cyclophyllideans

Our knowledge of parasites in wildlife remains limited, primarily due to restricted access to samples, especially of parasites from protected species. This present study contributes to the comprehension of the enigmatic world of helminths of African wild mammals and cestode biodiversity by combining both molecular and morphological analysis. Cestode samples were opportunistically collected from 77 individual definitive hosts in South Africa, Namibia and Ethiopia, encompassing 15 different species of wild African carnivores and additionally domestic cats. The analysis revealed 32 different cyclophyllidean species of which 21 (65.6 %) represent previously unknown genetic entities. They belong to the families Mesocestoididae, Hymenolepididae, Dipylidiidae and Taeniidae. Here we cover the non-taeniid cestodes, while the taeniids will be addressed in a separate publication. Three of the non-taeniid species uncovered in this study could be assigned to the genus Mesocestoides and were isolated from servals and domestic cats. The white-tailed mongoose was found to be a suitable host for a species belonging to the Hymenolepididae, which was identified as Pseudandrya cf. mkuzii. Both feline and canine genotypes of Dipylidium caninum were detected in domestic cats, the canine genotype also in an African wolf. In addition to these, a novel species of Dipylidium was discovered in an aardwolf. Lastly, four distinct species of Joyeuxiella were found in this study, revealing a cryptic species complex and emphasizing the need for a taxonomic reassessment of this genus. Despite the limited scope of our study in terms of geography and sample size, the results highlight that biodiversity of cestodes in African wild mammals is grossly under-researched and follow-up studies are urgently required, in particular linking morphology to gene sequences.

Diphyllobothrium sp. and Other Parasites of Migrating and Rare Fish Species in the Southern Baltic Sea and Coastal Waters, Poland.

The present study is the first to analyse the parasite fauna of sole Solea solea, dab Limanda limanda, hake Merluccius merluccius, whiting Merlangius merlangus, and plaice Pleuronectes platessa in the Pomeranian Bay, as well as saithe Pollachius virens from the Szczecin Lagoon (Poland). The aim of this study was to determine the occurrence of parasites in migrating and rare fish in the Pomeranian Bay and the Szczecin Lagoon and to determine the composition of the diet of these fish. The fish for analysis were obtained in the years 2010-2019. The typical marine nematode Capillaria (Procapillaria) gracilis, rarely recorded in Poland, was found, in addition to the following parasites: Ichthyophonus hoferi, Trichodina jadranica, Diphyllobothrium sp., Dichelyne (Cucullanellus) minutus, Raphidascaris acus, Anisakis simplex, Contracaecum osculatum, Hysterothylacium aduncum, Pseudoterranova decipiens, and Echinorhynchus gadi. Because the fish species analysed in the study are not typically present in the Baltic (with the exception of plaice), and because we do not know how long they feed while they are in the Baltic, we cannot be certain which parasites they acquired in the water bodies analysed in the study and which were introduced during the migration of fish. Although these fish are outside of their normal geographic range (except for plaice), in the new environment, there were enough suitable intermediate hosts for the parasites of these fish to complete their life cycle and survive.