Search

The genus Camellia contains three types of domesticates that meet various needs of ancient humans: the ornamental C. japonica, the edible oil-producing C. oleifera, and the beverage-purposed tea plant C. sinensis. The genomic drivers of the functional diversification of Camellia domesticates remain unknown. Here, we present the genomic variations of 625 Camellia accessions based on a new genome assembly of C. sinensis var. assamica ('YK10'), which consists of 15 pseudo-chromosomes with a total length of 3.35 Gb and a contig N50 of 816,948 bp. These accessions were mainly distributed in East Asia, South Asia, Southeast Asia, and Africa. We profiled the population and subpopulation structure in tea tree Camellia to find new evidence for the parallel domestication of C. sinensis var. assamica (CSA) and C. sinensis var. sinensis (CSS). We also identified candidate genes associated with traits differentiating CSA, CSS, oilseed Camellia, and ornamental Camellia cultivars. Our results provide a unique global view of the genetic diversification of Camellia domesticates and provide valuable resources for ongoing functional and molecular breeding research. [ABSTRACT FROM AUTHOR]

Introduction: Among cultivated tea plants (Camellia sinensis), only four mitogenomes for C. sinensis var. assamica (CSA) have been reported so far but none for C. sinensis var. sinensis (CSS). Here, two mitogenomes of CSS (CSSDHP and CSSRG) have been sequenced and assembled. Methods: Using a combination of Illumina and Nanopore data for the first time. Comparison between CSS and CSA mitogenomes revealed a huge heterogeneity. Results: The number of the repetitive sequences was proportional to the mitogenome size and the repetitive sequences dominated the intracellular gene transfer segments (accounting for 88.7%- 92.8% of the total length). Predictive RNA editing analysis revealed that there might be significant editing in NADH dehydrogenase subunit transcripts. Codon preference analysis showed a tendency to favor A/T bases and T was used more frequently at the third base of the codon. ENc plots analysis showed that the natural selection play an important role in shaping the codon usage bias, and Ka/Ks ratios analysis indicated Nad1 and Sdh3 genes may have undergone positive selection. Further, phylogenetic analysis shows that six C. sinensis clustered together, with the CSA and CSS forming two distinct branches, suggesting two different evolutionary pathway. Discussion: Altogether, this investigation provided an insight into evolution and phylogeny relationship of C. sinensis mitogenome, thereby enhancing comprehension of the evolutionary patterns within C. sinensis species. [ABSTRACT FROM AUTHOR]

Background: Since 2022, diagnostic laboratories in British Columbia have been advised to replace microscopy for intestinal ova and parasites with infectious diarrhea panel nucleic-acid amplification testing (IDP-NAAT). However, this multiplex assay captures only four common parasites: Cyclospora cayetanensis, Cryptosporidium spp., Entamoeba histolytica, and Giardia spp. Methods: An audit was conducted from September 2022 to August 2023, 1 year prior to implementation of IDP-NAAT in LifeLabs BC, when microscopy was the method used to identify intestinal parasites. Results: Pathogenic parasites were identified in 6149 of 52 221 stool specimens. The most common pathogens were Blastocystis hominis (78.47%), Dientamoeba fragilis (12.23%), Giardia spp. (4.93%), and Cyclospora spp. (1.07%). Entamoeba histolytica/dispar, Strongyloides stercoralis, Cryptosporidium spp., Ascaris lumbricoides, Diphyllobothrium spp., Enterobius vermicularis, Hymenolepis nana, Schistosoma mansoni, Taenia spp., Trichuris trichiura, and Clonorchis sinensis each accounted for less than 1% of the pathogenic parasites identified. Enterobius vermicularis was also identified in 46 of 1569 pinworm paddle specimens. Conclusions: Several potentially pathogenic parasites could be missed if only IDP-NAAT is used to detect intestinal parasites. If indicated, microscopy orders would be needed to capture parasites not detected by IDP-NAAT. [ABSTRACT FROM AUTHOR]

Copyright of Baghdad Science Journal is the property of Republic of Iraq Ministry of Higher Education & Scientific Research (MOHESR) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Enolase is a 47 kDa enzyme that functions within the glycolysis and gluconeogenesis pathways involved in the reversible conversion of D-2-phosphoglycerate (2PGA) to phosphoenolpyruvate (PEP). However, in the context of host-pathogen interactions, enolase from different species of parasites, fungi and bacteria have been shown to contribute to adhesion processes by binding to proteins of the host extracellular matrix (ECM), such as fibronectin (FN) or laminin (LM). In addition, enolase is a plasminogen (PLG)-binding protein and induces its activation to plasmin, the main protease of the host fibrinolytic system. These secondary 'moonlighting' functions of enolase are suggested to facilitate pathogen migration through host tissues. This study aims to uncover the moonlighting role of enolase from the parasite Fasciola hepatica, shedding light on its relevance to host-parasite interactions in fasciolosis, a global zoonotic disease of increasing concern. A purified recombinant form of F. hepatica enolase (rFhENO), functioning as an active homodimeric glycolytic enzyme of ~94 kDa, was successfully obtained, fulfilling its canonical role. Immunoblotting studies on adult worm extracts showed that the enzyme is present in the tegument and the excretory/secretory products of the parasite, which supports its key role at the host-parasite interface. Confocal immunolocalisation studies of the protein in newly excysted juveniles and adult worms also localised its expression within the parasite tegument. Finally, we showed by ELISA that rFhENO can act as a parasitic adhesin by binding host LM, but not FN. rFhENO also binds PLG and enhances its conversion to plasmin in the presence of the tissue-type and urokinase-type PLG activators (t-PA and u-PA). This moonlighting adhesion-like function of the glycolytic protein enolase could contribute to the mechanisms by which F. hepatica efficiently invades and migrates within its host and encourages further research efforts that are designed to impediment this function by vaccination or drug design. Author summary: Fasciola hepatica is a parasitic worm causing fasciolosis, primarily affecting herbivorous mammals and posing a significant veterinary problem. Furthermore, it is a zoonosis, meaning it can be transmitted to humans. F. hepatica enters the definitive host through ingestion of contaminated aquatic plants, migrating through the intestine to settle in the liver bile ducts, where it matures into the adult stage. To migrate, it utilizes various invasion strategies, including the use of multifunctional proteins, known as 'moonlighting'. In this study, we produced and molecularly characterized the parasitic enzyme enolase as a moonlighting protein to understand F. hepatica invasion mechanisms. We produced recombinant enolase with glycolytic activity, its canonical function in parasite energy production. Additionally, we localised this enzyme in the parasite's tegument, in direct contact with the host, and studied its ability to elicit an immune response early in ovine infection. Finally, we demonstrated the ability of enolase to interact with the extracellular matrix and the host's fibrinolysis, a proteolytic system responsible for dissolving blood clots. These secondary functions of F. hepatica enolase, described here for the first time, along with its localisation, suggest this protein as an interesting antigen for fasciolosis control. [ABSTRACT FROM AUTHOR]

In this study, we conducted tests on the seeds from four Taiwanese native Camellia species (C. japonica, C. furfuracea, C. laufoshanensis, and C. formosensis) and three commercialized species (C. oleifera, C. brevistyla, and C. sinensis) for comparison. We examined various aspects of these species, such as seed oil content, suitability for mechanical pressing, volatile components (edible flavor), and oil stability (suitability for cooking), to assess the feasibility of using these four native Taiwanese Camellia seeds as sources of edible oil. The results from solvent extraction tests and mechanical pressing experiments confirm that the seeds from C. furfuracea, C. japonica, and C. laufoshanensis have high oil contents, and their oils are suitable for extraction via the popular mechanical pressing method, with oil yields comparable to or higher than those of the commercialized Camellia species. The volatile components of the oils were collected using MonoTrap adsorbents and analyzed with a thermal desorption system coupled with gas chromatography–mass spectrometry (ATD-GC/MS), primarily consisting of alcohols, ketones, and aldehydes. The results of oxidative stability tests reveal that the seed oils from C. japonica, C. furfuracea, and C. laufoshanensis are higher than or equally stable to those from the commercialized Camellia species. After six months of storage, the stability of these three Camellia seed oils remained relatively high, demonstrating that the seed oils from C. japonica, C. furfuracea, and C. laufoshanensis can withstand high temperatures and can be easily preserved for future applications. [ABSTRACT FROM AUTHOR]

Background: Abiotic stress, such as salinity, affects the photosynthetic apparatus of plants. It is reported that the use of selenium nanoparticles (Se NPs), and biochemical compounds such as chitosan (CS) increase the tolerance of plants to stress conditions. Therefore, this study aimed to elucidate the potential of Se NPs, CS, and their composite (CS + Se NPs) in improving the photosynthetic apparatus of C. sinensis under salt stress in greenhouse conditions. The grafted seedlings of C. sinensis cv. Valencia after adapting to the greenhouse condition, were imposed with 0, 50, and 100 mM NaCl. After two weeks, the plants were foliar sprayed with distilled water (control), CS (0.1% w/v), Se NPs (20 mg L− 1), and CS + Se NPs (10 and 20 mg L− 1). Three months after treatment, the levels of photosynthetic pigments, leaf gas exchange, and chlorophyll fluorescence in the treated plants were evaluated. Results: Under salinity stress, total chlorophyll, carotenoid, and SPAD values decreased by 31%, 48%, and 28% respectively, and Fv/Fm also decreased compared to the control, while the ratio of absorption flux (ABS), dissipated energy flux (DI0) and maximal trapping rate of PSII (TR0) to RC (a measure of PSII apparent antenna size) were increased. Under moderate (50 mM NaCl) and intense (100 mM NaCl) salinity stress, the application of CS + Se NPs significantly increased the levels of photosynthetic pigments and the Fv/Fm value compared to plants treated with distilled water. Conclusions: It may be inferred that foliar treatment with CS + Se NPs can sustain the photosynthetic ability of C. sinensis under salinity stress and minimize its deleterious effects on photosynthesis. [ABSTRACT FROM AUTHOR]

Background and Aim: Heat stress (HS) can negatively impact oocytes by disrupting mitochondrial activity, increasing the production of reactive oxygen species, and decreasing antioxidant levels. This study investigated the impact of orange peel (OP) exposure on zebrafish oocytes (ZOs) diameter, survival rate, and germinal vesicle breakdown (GVBD) during HS. Material and Methods: We investigated the antioxidant effect of flavonoids (concentration = 328.58 ppm) derived from OP (Citrus sinensis) added to in vitro maturation (IVM) media of ZOs (Danio rerio) under non-heat stress (NHS) and HS conditions to mimic in vivo HS conditions due to the global warming phenomenon on females. ZO in stage 3 (n = 1080) was treated with 4 µL of OP extract (not treated/control) under HS: 32°C (Heat stress 32°C solution/Heat stress 32°C orange peel [HS32S/HS32O]) and 34°C (Heat stress 34°C solution/Heat stress 34°C orange peel [HS34S/HS34O]); and NHS: 28°C (Non-heat stress solution/Non-heat stress orange peel [NHSS/NHSO]), during maturation. After 24 h of maturation, we observed the oocyte diameter, survival rate, and GVBD rate. The data were analyzed with IBM Statistics 23 software using two-way analysis of variance and Kruskal--Wallis (p < 0.05). Results: The highest oocyte diameter data were in NHS treated with OP extract (NHSO) group (0.759 ± 0.01; mean ± standard error) compared with HS group using and without OP extract (HS32S [0.583 ± 0.02]; HS32O [0.689 ± 0.02]; HS34S [0.554 ± 0.02]; and HS34O [0.604 ± 0.02]). The survival rate of OP treated group, namely, NHSO (93% ± 3%), HS32O (85% ± 2%), and HS34O (80% ± 2%) was higher than that of the group without treatment (NHSS [83% ± 3%], HS32S [71% ± 6%], and HS34S [63% ± 3%]). ZO treated with OP extract (NHSO [93% ± 3%], HS32O [85% ± 2%], and HS34O [80% ± 2%]) showed a higher GVBD rate than the group without treatment (NHSS [83% ± 3%], HS32S [71% ± 6%], and HS34S [63% ± 3%]). Conclusion: It revealed that OP can enhance the oocyte diameter, survival rate, and GVBD rate of ZO under NHS and HS. Further investigation should be conducted to determine the effect of OP extract (C. sinensis) on in vivo conditions in females as an alternative treatment to face global warming. [ABSTRACT FROM AUTHOR]

Simple Summary: The natural Cordyceps sinensis insect–fungal complex is one of the most highly valued therapeutic agents in traditional Chinese medicine. The sexual reproduction of Ophiocordyceps sinensis is controlled by the expression of mating-type genes within the MAT1-1 and MAT1-2 idiomorphs and a-/α-pheromone receptor genes. This study revealed the differential expression of these genes in Hirsutella sinensis (Genotype #1 among the 17 genotypes of O. sinensis) and alternative splicing of the MAT1-2-1 gene, characterized by an unspliced intron I that contains stop codons. Mutations of the mating and pheromone receptor proteins were also identified, potentially leading to changes in the secondary structures and functionalities of the proteins. These results indicate the requirement of mating partners under either heterothallism or hybridization during the sexual life of natural C. sinensis but do not support the "self-fertilization" hypothesis for O. sinensis under homothallism or pseudohomothallism. Sexual reproduction in ascomycetes is controlled by the mating-type (MAT) locus. (Pseudo)homothallic reproduction has been hypothesized on the basis of genetic data from Hirsutella sinensis (Genotype #1 of Ophiocordyceps sinensis). However, the differential occurrence and differential transcription of mating-type genes in the MAT1-1 and MAT1-2 idiomorphs were found in the genome and transcriptome assemblies of H. sinensis, and the introns of the MAT1-2-1 transcript were alternatively spliced with an unspliced intron I that contains stop codons. These findings reveal that O. sinensis reproduction is controlled at the genetic, transcriptional, and coupled transcriptional-translational levels. This study revealed that mutant mating proteins could potentially have various secondary structures. Differential occurrence and transcription of the a-/α-pheromone receptor genes were also found in H. sinensis. The data were inconsistent with self-fertilization under (pseudo)homothallism but suggest the self-sterility of H. sinensis and the requirement of mating partners to achieve O. sinensis sexual outcrossing under heterothallism or hybridization. Although consistent occurrence and transcription of the mating-type genes of both the MAT1-1 and MAT1-2 idiomorphs have been reported in natural and cultivated Cordyceps sinensis insect-fungi complexes, the mutant MAT1-1-1 and α-pheromone receptor transcripts in natural C. sinensis result in N-terminal or middle-truncated proteins with significantly altered overall hydrophobicity and secondary structures of the proteins, suggesting heterogeneous fungal source(s) of the proteins and hybridization reproduction because of the co-occurrence of multiple genomically independent genotypes of O. sinensis and >90 fungal species in natural C. sinensis. [ABSTRACT FROM AUTHOR]

Simple Summary: Our study investigated the molecular mechanism of co-cultivated Citrus sinensis in diminishing the resistance of Poncirus trifoliata to the Asian Citrus Psyllid (ACP). The presence of C. sinensis was found to weaken the resistance of P. trifoliata to ACP, shown by a significant increase in ACP survival after 14 days of incubation. RNA-seq analysis revealed differentially expressed genes (DEGs) between control samples and those under co-cultivation with C. sinensis, with significant changes in metabolic processes. Further analysis indicated that P. trifoliata co-cultivation with C. sinensis significantly impacted the expression of many genes, including those involved in polysaccharide metabolic processes, metabolic pathways, biosynthesis of secondary metabolites, and phenylpropanoid biosynthesis. The study also identified a hub gene, Ptrif_0004s_1029, a NAC gene from transcription factor genes, appearing crucial in transcriptional regulation related to potential ACP resistance. Additionally, DEGs related to ACP feeding were also identified. Our research has elucidated the molecular mechanisms that underlie the resistance of P. trifoliata to the ACP and identified potential genes associated with this resistance. Citrus huanglongbing (HLB) is a devastating disease for citrus production, largely caused by the Asian citrus psyllid (ACP). Poncirus trifoliata exhibits high resistance to ACP; however, this resistance is weakened when C. sinensis is co-cultivated. This study aimed to identify the differentially expressed genes (DEGs) during ACP feeding and to uncover potential ACP resistance genes in P. trifoliata. In comparison to independent cultivation, 1247 and 205 DEGs were identified in P. trifoliata when co-cultivated with C. sinensis after 7 and 14 days, respectively. Analysis of enriched Gene Ontology categories revealed that DEGs were significantly associated with the cell wall, glucometabolic activities, and secondary metabolites. Additionally, these genes were found to be involved in phytohormone signaling, cell wall metabolism, redox state homeostasis, and secondary metabolites, as well as a number of transcription factor genes (TFs). Furthermore, we examined the impact of the ACP feeding factor on the gene expression patterns in P. trifoliata. Results showed an increase in the JA signaling pathway and various TFs. The RNA-seq results were verified using reverse transcription quantitative PCR. Our findings shed light on the molecular basis of ACP resistance in P. trifoliata and identified potential genes associated with this resistance. [ABSTRACT FROM AUTHOR]

Dwarfing is a pivotal agronomic trait affecting both yield and quality. Citrus species exhibit substantial variation in plant height, among which internode length is a core element. However, the molecular mechanism governing internode elongation remains unclear. Here, we unveiled that the transcriptional cascade consisting of B‐BOX DOMAIN PROTEIN 22 (BBX22) and ELONGATED HYPOCOTYL 5 (HY5) finely tunes plant height and internode elongation in citrus. Loss‐of‐function mutations of BBX22 in an early‐flowering citrus (Citrus hindsii "SJG") promoted internode elongation and reduced pigment accumulation, whereas ectopic expression of BBX22 in SJG, sweet orange (C. sinensis), pomelo (C. maxima) or heterologous expression of BBX22 in tomato (Solanum lycopersicum) significantly decreased internode length. Furthermore, exogenous application of gibberellin A3 (GA3) rescued the shortened internode and dwarf phenotype caused by BBX22 overexpression. Additional experiments revealed that BBX22 played a dual role in regulation internode elongation and pigmentation in citrus. On the one hand, it directly bound to and activated the expression of HY5, GA metabolism gene (GA2 OXIDASE 8, GA2ox8), carotenoid biosynthesis gene (PHYTOENE SYNTHASE 1, PSY1) and anthocyanin regulatory gene (Ruby1, a MYB DOMAIN PROTEIN). On the other hand, it acted as a cofactor of HY5, enhancing the ability of HY5 to regulate target genes expression. Together, our results reveal the critical role of the transcriptional cascade consisting of BBX22 and HY5 in controlling internode elongation and pigment accumulation in citrus. Unraveling the crosstalk regulatory mechanism between internode elongation and fruit pigmentation provides key genes for breeding of novel types with both dwarf and health‐beneficial fortification in citrus. [ABSTRACT FROM AUTHOR]

We developed a new concentration kit, called the ParaEgg (PE), for easy detection trematode eggs from fecal samples in endemic areas of clonorchiasis and metagonimiasis in Korea. To create a standard of detection efficiency, 120 fecal samples were examined using the water–ether concentration method (WECM). The PE kit and Mini ParaSep (PS) kit were used to compare the detection sensitivity of 100 egg-positive and 20 egg-negative samples in WECM. Additionally, stool samples, which were intentionally spiked with 10, 20, and 30 Clonorchis sinensis eggs, were evaluated to assess the sensitivity in low-infection cases. The PE and PS kits showed detection rates of 100% and 92%, respectively, from 100 egg-positive samples in WECM. Meanwhile, eggs were detected in 3 (PE) and 2 (PS) out of 20 egg-negative samples in WECM. The PE kit detected the highest number of eggs per gram of feces (727 on average), followed by the WECM (524) and PS kit (432). In fecal samples that were intentionally spiked with 10, 20, and 30 C. sinensis eggs, PE only detected eggs 2 out of 5 samples in 10 eggs spiked (40%), and the detection rates were 80% and 100%, respectively. The PE kit enabled a more accurate identification of trematode eggs because of the clearance of small fecal debris in the microscopic field. In conclusion, the PE kit is obviously helpful to detect and identify trematode eggs in stool examinations especially in endemic areas of clonorchiasis and metagonimiasis. [ABSTRACT FROM AUTHOR]

Since the last century, it has been shown that dedifferentiated cells of Camellia sinensis can produce catechins and other secondary metabolites under in vitro conditions, with potential applications in the cosmetic, pharmaceutical and food industries. In this work, cell suspension cultures of a C. sinensis cell line (LSC-5Y) were established in a liquid medium in order to optimize the biomass productivity, catechin monomer (GC, EGC, C, EC, CG, and ECG) and alkaloid (TB and CAF) productivity. The following factors were evaluated: concentration of growth regulators (BA and IBA), inoculum size, age of the cell line, light exposure, and effect of biotic elicitors (MeJA and extracts of Ciborinia camelliae). GC, EGC, and ECG increased approximately 1.80-fold when the auxin IBA concentration was increased from 0.1 to 2.0 mg/L. In addition, better productivity of EGC, C, EC, and CAF was achieved by using inoculum densities between 50 and 100 g/L. Although lower inoculum densities (25 g/L) showed a higher growth rate (0.20 d−1), the use of inoculum densities higher than 25 g/L favors a 2–4-fold increase in total catechin (TC) productivity, with maximum productivity being reached after 21 days of culture. However, the cell line showed instability in TC productivity: in the short term (in three successive subcultures), the coefficient of variation was 32.80%, and catechin production capacity was 2.5 years with maximum productivity at 0.5 years. Finally, it was observed that ethanol, used as an elicitor solvent, has a strong elicitor effect capable of increasing the accumulation of catechins up to 5.24 times compared to the treatment without an elicitor. [ABSTRACT FROM AUTHOR]

Opisthorchis viverrini is the etiological agent of the disease opisthorchiasis and related cholangiocarcinoma (CCA). It infects fish-eating mammals and more than 10 million people in Southeast Asia suffered from opisthorchiasis with a high fatality rate. The only effective drug against this parasite is Praziquantel, which has significant side effects. Due to the lack of appropriate treatment options and the high death rate, there is a dire need to develop novel therapies against this pathogen. In this study, we designed a multi-epitope chimeric vaccine design against O. viverrini by using immunoinformatics approaches. Non-allergenic and immunogenic MHC-1, MHC-2, and B cell epitopes of three candidate proteins thioredoxin peroxidase (Ov-TPx-1), cathepsin F1 (Ov-CF-1) and calreticulin (Ov-CALR) of O. viverrini, were predicted to construct a potent multiepitope vaccine. The coverage of the HLA-alleles of these selected epitopes was determined globally. Four vaccine constructs made by different adjuvants and linkers were evaluated in the context of their physicochemical properties, antigenicity, and allergenicity. Protein-protein docking and MD simulation found that vaccines 3 was more stable and had a higher binding affinity for TLR2 and TLR4 immune receptors. In-silico restriction cloning of vaccine model led to the formation of plasmid constructs for expression in a suitable host. Finally, the immune simulation showed strong immunological reactions to the engineered vaccine. These findings suggest that the final vaccine construct has the potential to be validated by in vivo and in vitro experiments to confirm its efficacy against the CCA causing O. viverrini. [ABSTRACT FROM AUTHOR]

As an important genus in Orchidaceae, Cymbidium has rich ecological diversity and significant economic value. DNA binding with one zinc finger (Dof) proteins are pivotal plant-specific transcription factors that play crucial roles in the growth, development, and stress response of plants. Although the Dof genes have been identified and functionally analyzed in numerous plants, exploration in Orchidaceae remains limited. We conducted a thorough analysis of the Dof gene family in Cymbidium goeringii, C. ensifolium, and C. sinensis. In total, 91 Dof genes (27 CgDofs, 34 CeDofs, 30 CsDofs) were identified, and Dof genes were divided into five groups (I–V) based on phylogenetic analysis. All Dof proteins have motif 1 and motif 2 conserved domains and over half of the genes contained introns. Chromosomal localization and collinearity analysis of Dof genes revealed their evolutionary relationships and potential gene duplication events. Analysis of cis-elements in CgDofs, CeDofs, and CsDofs promoters showed that light-responsive cis-elements were the most common, followed by hormone-responsive elements, plant growth-related elements, and abiotic stress response elements. Dof proteins in three Cymbidium species primarily exhibit a random coil structure, while homology modeling exhibited significant similarity. In addition, RT-qPCR analysis showed that the expression levels of nine CgDofs changed greatly under heat stress. CgDof03, CgDof22, CgDof27, CgDof08, and CgDof23 showed varying degrees of upregulation. Most upregulated genes under heat stress belong to group I, indicating that the Dof genes in group I have great potential for high-temperature resistance. In conclusion, our study systematically demonstrated the molecular characteristics of Dof genes in different Cymbidium species, preliminarily revealed the patterns of heat stress, and provided a reference for further exploration of stress breeding in orchids. [ABSTRACT FROM AUTHOR]

Pickled tea is an anaerobically fermented tea common in Thailand, Myanmar and Yunnan minority areas. γ-aminobutyric acid (GABA) is non-protein amino acid with multiple bioactives, which can be easily produced under anaerobic conditions. During the processing of pickled tea, controlling the process parameters is effective for the production of GABA-rich products; however, the precise parameters remain to be clarified. In the present study, the fresh leaves of Camellia sinensis (L.) Kuntze (C. sinensis) 'FudingDabai', C. sinensis 'MabianLv No. 1', C. sinensis 'Wuniuzao' and C. sinensis 'Fuxuan No. 9' were used as raw materials to process GABA-rich pickled tea. Single-factor and orthogonal experiments were conducted to determine the best tea cultivars and optimize the best processing parameters via comparing the content of GABA, tea polyphenols (TPs) and other biochemical components of GABA-rich pickled tea. The results of the signal-factor experiment showed that the fresh leaves of C. sinensis 'MabianLv No. 1' had the highest GABA content of 2.61 mg·g−1 after treatment with vacuum for 6 h; therefore, C. sinensis 'MabianLv No. 1' was selected as the raw material for the subsequent experiments. Orthogonal experiments showed that the highest GABA content of 2.53 mg·g−1 was found in the pickled tea with 8 h of vacuum treatment, 20 min of rolling after microwave fixing, 20 min of spreading and 20 d of anaerobic fermentation at room temperature. Further, the sensory evaluation showed that it possesses a strong sour taste with a slight sweetness and a light yellow color and better comprehensive quality. This indicates that these parameters are optimal for the processing of GABA-rich pickled tea. This study will provide scientific basis for the subsequent production of high GABA tea. [ABSTRACT FROM AUTHOR]

Clonorchis sinensis is a significant zoonotic food-borne parasite that causes a range of hepatobiliary diseases, which in severe cases can even lead to cholangiocarcinoma. To explore new diagnostic and treatment strategies, the dynamic RNA regulatory processes across different developmental stages of C. sinensis were analyzed by using whole-transcriptome sequencing. The chromosomal-level genome of C. sinensis was used for sequence alignment and annotation. In this study, we identified a total of 59,103 RNAs in the whole genome, including 2,384 miRNAs, 25,459 mRNAs, 27,564 lncRNAs and 3,696 circRNAs. Differential expression analysis identified 6,556 differentially expressed mRNAs, 2,231 lncRNAs, 877 miRNAs and 20 circRNAs at different developmental stages. Functional enrichment analysis highlighted the critical role of microtubule-related biological processes in the growth and development of C. sinensis. And coexpression analysis revealed 97 lncRNAs and 85 circRNAs that were coexpressed with 42 differentially expressed mRNAs that associated with microtubules at different developmental stages of C. sinensis. The expression of the microtubule-related genes dynein light chain 2 (DLC2) and dynein light chain 4 (DLC4) increased with C. sinensis development, and DLC2/4 could be inhibited by albendazole. Finally, by constructing competing endogenous RNA (ceRNA) networks, the lncRNA-miRNA-mRNA and circRNA-miRNA-mRNA regulatory relationships were constructed, and the ceRNA networks of MSTRG.14258.5-novel_miR_2287-newGene_28215 and MSTRG.14258.5-novel_miR_2216-CSKR_109340 were verified. This study suggests, through whole transcriptome sequencing, that the context of microtubule regulation may play an essential role in the development and growth of C. sinensis. Author summary: This study is the first to comprehensively elucidate the entire transcriptome of Clonorchis sinensis by exploring the expression and regulatory dynamics of noncoding RNAs and mRNAs at different developmental stages in its life cycle. By using next-generation sequencing technology, this research demonstrated new mRNAs, miRNAs, lncRNAs and circRNAs, and established valuable resources for future parasitic research. Compared with mRNAs, lncRNAs have shorter gene lengths, lower expression levels, fewer exons and fewer isoforms. Differential expression analysis demonstrated stage-specific patterns of noncoding RNAs and mRNAs in C. sinensis during its life cycle, thus providing crucial insights into the role of noncoding RNAs in parasite growth and development. Furthermore, the construction of ceRNA regulatory networks revealed that microtubule-related lncRNAs and circRNAs in C. sinensis may play an important role in its growth and development. This study advances our understanding of the entire transcriptome of C. sinensis and introduces novel perspectives for future parasitic infection treatment strategies. In summary, this study establishes a foundation for in-depth investigations into the interplay between noncoding RNAs and parasites, as well as the associated biological processes, thus demonstrating broad research and therapeutic potential. [ABSTRACT FROM AUTHOR]

Background: The liver fluke Clonorchis sinensis imports large amounts of glucose to generate energy and metabolic intermediates through glycolysis. We hypothesized that C. sinensis absorbs glucose through glucose transporters and identified four subtypes of glucose transporter (CsGTP) and one sodium glucose co-transporter (CsSGLT) in C. sinensis. Methodology/Principal findings: Expressed sequence tags encoding CsGTPs were retrieved from the C. sinensis transcriptome database, and their full-length cDNA sequences were obtained by rapid amplification of cDNA ends (RACE). The tissue distribution of glucose transporters in C. sinensis adults was determined using immunohistochemical staining. Developmental expression was measured using RT-qPCR. The transport and distribution of glucose into living C. sinensis were monitored using confocal microscopy. Membrane topology and key functional residues of CsGTPs were homologous to their counterparts in animals and humans. CsGTP1, 2, and 4 were transcribed 2.4–5.5 times higher in the adults than metacercariae, while CsGTP3 was transcribed 2.1 times higher in the metacercariae than adults. CsSGLT transcription was 163.6 times higher in adults than in metacercariae. In adults, CsSGLT was most abundant in the tegument; CsGTP3 and CsSGLT were localized in the vitelline gland, uterine wall, eggs, mesenchymal tissue, and testes; CsGTP4 was found in sperm and mesenchymal tissue; and CsGTP1 was mainly in the sperm and testes. In C. sinensis adults, exogenous glucose is imported in a short time and is present mainly in the middle and posterior body, in which the somatic and reproductive organs are located. Of the exogenous glucose, 53.6% was imported through CsSGLT and 46.4% through CsGTPs. Exogenous glucose import was effectively inhibited by cytochalasin B and phlorizin. Conclusions/Significance: We propose that CsSGLT cooperates with CsGTPs to import exogenous glucose from the environmental bile, transport glucose across mesenchymal tissue cells, and finally supply energy-demanding organs in C. sinensis adults. Studies on glucose transporters may pave the way for the development of new anthelmintic drugs. Author summary: The Chinese liver fluke C. sinensis is a fish-borne trematode that causes serious biliary diseases and complications, such as cholangiocarcinoma. Living in an anaerobic environment, C. sinensis adults consume large amounts of exogenous glucose and produce energy and intermediate products via glycolytic pathways. We identified four glucose transporters (CsGTP1–4) and one sodium glucose co-transporter (CsSGLT) in C. sinensis and their glucose transport mechanism was investigated. This is the first study to report SGLT in a helminthic parasite, particularly trematodes. Elucidating the structure and function of CsGTPs, CsSGLT, and their inhibitors may lead to strategies to exhaust glucose and ultimately starve C. sinensis. This study provides fundamental information on the glucose transporters of helminths and may shed light on directions for studying energy metabolism in other mammalian-infecting trematodes. [ABSTRACT FROM AUTHOR]

Citrus is commercially propagated via grafting, which ensures trees have consistent fruit traits combined with favorable traits from the rootstock such as soil adaptability, vigor, and resistance to soil pathogens. Graft incompatibility can occur when the scion and rootstock are not able to form a permanent, healthy union. Understanding and preventing graft incompatibility is of great importance in the breeding of new fruit cultivars and in the choice of scion and rootstock by growers. The rootstock US-1283, a citrandarin generated from a cross of "Ninkat" mandarin (Citrus reticulata) and "Gotha Road" #6 trifoliate orange (Poncirus trifoliata), was released after years of field evaluation because of its superior productivity and good fruit quality on "Hamlin" sweet orange (C. sinensis) under Florida's growing conditions. Subsequently, it was observed that trees of "Bearss" lemon (C. limon) and "Valencia" sweet orange (C. sinensis) grafted onto US-1283 exhibited unhealthy growth near the graft union. The incompatibility manifested as stem grooving and necrosis underneath the bark on the rootstock side of the graft. Another citrandarin rootstock, US-812 (C. reticulata "Sunki" x P. trifoliata "Benecke"), is fully graft compatible with the same scions. Transcriptome analysis was performed on the vascular tissues above and below the graft union of US- 812 and US-1283 graft combinations with "Bearss" and "Valencia" to identify expression networks associated with incompatibility and help understand the processes and potential causes of incompatibility. Transcriptional reprogramming was stronger in the incompatible rootstock than in the grafted scions. Differentially expressed genes (DEGs) in US-1283, but not the scions, were associated with oxidative stress and plant defense, among others, similar to a pathogen-induced immune response localized to the rootstock; however, no pathogen infection was detected. Therefore, it is hypothesized that this response could have been triggered by signaling miscommunications between rootstock and scion either through (1) unknown molecules from the scion that were perceived as danger signals by the rootstock, (2) missing signals from the scion or missing receptors in the rootstock necessary for the formation of a healthy graft union, (3) the overall perception of the scion by the rootstock as non-self, or (4) a combination of the above. [ABSTRACT FROM AUTHOR]

An ample amount of fruit waste is generated as agro-industrial waste, leading to significant nutritional, economic, and environmental challenges. Fruit peels are rich in many valuable bioactive compounds with the potential for developing nanoparticles. This study examined fresh juices of two citrus fruit peel wastes (Citrus sinensis: C. sinesis and Citrus limon: C. limon) for antioxidants and total protein. Then, we investigated their ability to produce silver nanoparticles, which were further analyzed for anti-microbial activity against thirteen pathogenic microbes. Both Citrus peel juices were rich in secondary metabolites. The formation of Ag nanoparticles was initially confirmed by UV-vis spectroscopy, with peaks at 400 nm for C. sinensis peel Ag nanoparticles and 430 nm for C. limon peel Ag nanoparticles. Further characterization was conducted using zeta sizer, zeta potential, Transmission Electron Microscopy (TEM), Scanning Electron Microscope (SEM), Energy-dispersive X-ray spectroscopy (EDX) and Fourier transform infrared (FTIR) spectroscopy. The antimicrobial activity was tested using the well diffusion method against 11 bacterial strains (five Gram-positive and six Gram-negative) and two fungal strains of Candida. TEM and SEM results revealed a spherical shape, with an average diameter of about 13 nm for C. sinensis and 21 nm for C. limon Ag. EDX confirmed the presence of silver in both nanoparticles. The FTIR spectrum of the extract indicated the presence of biomolecules, which facilitated the reduction and capping of the synthesized Ag nanoparticles. The prepared nanoparticles showed remarkable antimicrobial activity, but the nanoparticles from C. sinensis exhibited stronger antibacterial properties because of their smaller size. Citrus peel waste is a suitable medium for the eco-friendly production of silver nanoparticles. [ABSTRACT FROM AUTHOR]

Objective: This study assessed the cardioprotective properties of Persicaria maculosa (PME) and Citrus sinensis (CME) hydromethanolic extracts, besides Citrus sinensis aqueous extract (CWE) against doxorubicin (DOX)-induced cardiotoxicity. Materials and Methods: The extracts were characterized. Mice were divided into eight groups: control (saline), DOX, protected (injected with 200 mg/kg of PME, CWE or CME for 21 days, orally, and DOX), and extracts (PME, CWE or CME administration, orally, for 21 days). DOX was injected (5 mg/kg, ip) on days 8, 13 and 18 of the experiment. Cardiac tumor necrosis factor-alpha (TNF-a), nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and carbonyl reductase 1 (CBR1) expression levels, besides superoxide dismutase, catalase, malondialdehyde, nitric oxide and total protein levels were evaluated. Serum lactate dehydrogenase, creatine phosphokinase cardiac isoenzyme, aspartate transaminase, cholesterol, triglycerides and creatinine levels, as well as the cardiac tissues were examined. Results: Comparing with the control, DOX considerably (p<0.01) up-regulated TNF-a expression, malondialdehyde, nitric oxide, cardiac enzymes, lipids and creatinine levels, while it significantly (p<0.01) down-regulated Nrf2 and CBR1. Additionally, DOX interfered with antioxidant enzymes' activities (p<0.01). Conversely, protected groups showed a significant (p<0.01) amelioration of DOX-induced cardiotoxic effects. Conclusion: The current study provides a new understanding of P. maculosa and C. sinensis cardioprotective mechanisms. The extracts' cardioprotective effects may be due to their antioxidant activities, ability to maintain the redox homeostasis through regulation of important antioxidant genes and primary antioxidant enzymes, and capability to recover inflammatory cytokines and lipids levels. Noteworthy, the tested extracts showed no toxic changes on the normal mice. [ABSTRACT FROM AUTHOR]

Aim: The aim of this study was to evaluate the effect of proanthocyanidin and C. sinensis-polyphenols on microtensile bonding properties of prepared teeth with resin-modified glass ionomer cement (GIC). Setting and Design: This was an in vitro study. Materials and Methods: Seventy-eight maxillary premolars were selected and mounted into auto-polymerizing acrylic resin blocks. The samples were prepared and metal crowns were fabricated. The samples were randomly divided into three groups. Samples under Group 1 were not treated with any of the extracts and followed conventional bonding protocol. Samples under Group 2 and Group 3 were treated with proanthocyanidin and C. sinensis-polyphenols, respectively. After dentin treatment, these samples were luted to metal crowns using resin-modified GIC. Universal testing machine was used to measure the load at which the crowns were debonded and microtensile bond strength in MPa was calculated. Statistical Analysis: The results were statistically analyzed using one-way ANOVA and post hoc Tukey HSD. Results: Samples treated with C. sinensis polyphenols (Group 3) had maximum bond strength followed by Group 2, where the samples were treated with proanthocyanidin. Conclusion: C. sinensis polyphenols due to their anti-proteolytic and antioxidant properties showed improved bond strength compared to proanthocyanidin, a cross-linking agent, followed by conventional bonding protocol. [ABSTRACT FROM AUTHOR]

The initial valves of two Cymbella species are observed under a scanning electron microscope, and the perizonium ultrastructure of Cymbella is revealed for the first time. The perizonium is composed of alternate nodes and internodes and lacks transverse perizonium bands. Four new species, Cymbella apiculatophora sp. nov., C. hunanensis sp. nov., C. juglandis sp. nov. and C. menyuanensis sp. nov., are described using light and scanning electron microscopy based on epilithon samples collected from rivers in Hunan and Qinghai Provinces, China. Cymbella menyuanensis is a typical Cymbella species that closely resembles species in the group around C. cymbiformis Agardh, the type species of the genus. Cymbella apiculatophora is similar to C. sinensis Metzeltin & Krammer, while Cymbella hunanensis is closer to the C. hustedtii Krasske group. The last species, C. juglandis, has a cymbelloid valve outline, an obscured intermissio, internal occlusions of the areolae, dorsally deflected distal raphe fissures and a divided apical pore field at each apex, and it does not appear to belong to any group. In addition, new observations on C. cf. excisiformis Krammer and C. hustedtii are reported. The current concept of the genus Cymbella does not represent a monophyletic group as shown by molecular phylogenetic analyses. However, these analyses are still at the preliminary stage and are not yet sufficient to support a complete revision of the genus. Thus, although extremely diverse ultrastructural features are observed in the six Cymbella species investigated in this paper, we prefer to keep them within Cymbella at this moment for the sake of nomenclatural stability. [ABSTRACT FROM AUTHOR]

Clonorchis sinensis infections persist globally among humans. These pathogens mainly inhabit the intrahepatic biliary system. Most individuals with clonorchiasis exhibit mild symptoms. The absence of distinctive symptoms often results in delayed diagnosis and treatment, potentially leading to chronic infection. We herein report a case of a 29-year-old female presented with a year-long history of abdominal distention and dyspepsia. Imaging revealed intrahepatic bile duct dilatation, intrahepatic bile duct cyst, and associated deposits. One month post-cystectomy, the patient developed massive ascites and a significant increase in eosinophil count. After treatment, multiple worms were observed in the drainage tube. Morphological and DNA metagenomic analyses confirmed the presence of C. sinensis. Clinical manifestations of C. sinensis vary widely. Imaging serves as a valuable diagnostic tool in endemic areas, especially in detecting intrahepatic duct dilation where the flukes reside. In addition to intrahepatic bile duct dilation, abnormal echoes within the bile duct and the presence of floating objects in the gallbladder significantly aid in diagnosis. Clinicians may encounter these parasitic diseases unexpectedly, underscoring the importance of understating such cases in routine practice and contributing to our broader understanding of managing similar cases in clinical settings.

Abstract Background Opisthorchiasis and clonorchiasis, caused by Opisthorchis viverrini and Clonorchis sinensis, respectively, are significant yet neglected foodborne trematodiases in the Great Mekong Subregion (GMS). Despite the reporting of the prevalence of these human liver flukes in the region over the past decades, there has been a lack of a comprehensive and systematic consolidation of this data. Therefore, we aimed to conduct a thorough systematic review and meta-analysis to synthesize and analyze time-trend prevalence estimates of both O. viverrini and C. sinensis across the GMS for the past 30 years. Methods This study undertakes a systematic review using a comprehensive search for published articles in PubMed, EMBASE, Scopus, Cochrane and Thai Journal Online databases until early 2023. The pooled prevalence of O. viverrini and C. sinensis infection was analyzed through a random-effects meta-analysis, with meta-regression analysis used to quantify associations with study characteristics. Sub-group analysis was conducted, whenever comparison data were available, to assess the risk of O. viverrini and C. sinensis infection in each GMS country. Heterogeneity among studies was assessed using the Q statistic and quantified by using the I 2 Index. Results From a total of 2997 articles, 155 articles comprising 218 datasets and 751,108 participants were included for review. The GMS prevalence of O. viverrini was 21.11% [45,083/260,237; 95% confidence interval (CI): 17.74–24.47%]. Pooled prevalence estimates were highly observed in Laos (34.06%, 95% CI: 26.85–41.26%), followed by Thailand (18.19%, 95% CI: 13.86–22.51%), and Cambodia (10.48%, 95% CI: 5.52–15.45%). Myanmar and Vietnam had limited data sources for calculation. Clonorchis sinensis infection in GMS was 25.33% (95% CI: 18.32–32.34%), with Guangxi, China, exhibiting the highest prevalence rates at 26.89% (95% CI: 18.34–35.43%), while Vietnam had a prevalence rate of 20.30% (95% CI: 9.13–31.47%). O. viverrini prevalence decreased significantly over time, whereas C. sinensis infection appeared to be stable consistently over time in both China and Vietnam. Conclusions This comprehensive study, drawing from the largest datasets to date, offers an in-depth systematic prevalence review of human liver flukes in the Greater Mekong Subregion. It underscores the imperative for systematic surveillance, data collection, and the implementation of intervention and control measures for these infectious diseases of poverty. Graphical Abstract

The family Acuariidae is a speciose group of parasitic nematodes, infecting mostly birds as definitive hosts. This study focused on the characterization of two species of acuariids, collected in two different species of piscivorous birds, the European great cormorant Phalacrocorax carbo sinensis from Italy, and the pygmy cormorant Microcarbo pygmaeus from Israel. Parasites were analyzed using light and scanning electron microscopy and by amplification and sequencing of the 28S rDNA. The results of morphological and molecular analyses showed that Ph. carbo sinensis was infected by the acuariid Syncuaria squamata (12 females) and Cosmocephalus obvelatus (1 female), whereas M. pygmaeus was infected by C. obvelatus (2 males, 12 females). The present results provide new data on the distribution of acuariid parasites of piscivorous birds, the first report of Acuariidae in Israel, and the first molecular data on S. squamata and C. obvelatus, which will be useful in future epidemiological and phylogenetic studies of these widely distributed, but less molecularly studied parasites. [ABSTRACT FROM AUTHOR]

Bisphenol S (BPS), one of the substitutes for bisphenol A (BPA), is widely used in various commodities. The BPS concentrations in surface water have gradually increased in recent years, making it a predominant bisphenol analogue in the aquatic environment and raising concerns about its health and ecological effects on aquatic organisms. For this study, we conducted a 96 h acute toxicity test and a 15-day developmental exposure test to assess the adverse effects of BPS exposure in Chinese medaka (Oryzias sinensis), a new local aquatic animal model. The results indicate that the acute exposure of Chinese medaka embryos to BPS led to relatively low toxicity. However, developmental exposure to BPS was found to cause developmental abnormalities, such as decreased hatching rate and body length, at 15 dpf. A transcriptome analysis showed that exposure to different concentrations of bisphenol S often induced different reactions. In summary, environmental concentrations of BPS can have adverse effects on the hatching and physical development of Chinese medaka, and further attention needs to be paid to the potential toxicity of environmental BPS. [ABSTRACT FROM AUTHOR]

[Objective] Trehalase (Tre) is a key enzyme in trehalose metabolism of Conopomorpha sinensis Bradley, which plays an important role in energy metabolism and growth development by specifically hydrolyzing trehalose into glucose. The study aims to clone two trehalase genes (CsTre1 and CsTre2) from Conopomorpha sinensis Bradley, to clarify its expression patterns in different developmental stages and tissues, and to analyze the molecular characteristics of the two genes and their enzyme proteins.[Method] Based on the transcriptome data of C. sinensis, the full-length cDNA sequences of CsTre1 and CsTre2 were cloned with the rapid amplification of cDNA ends (RACE)-PCR. Bioinformatics analysis was performed with software such as ORF Finder, ProtParam, SignalP 4.1, ProtScale, NetPhos2.0 Server and IQ TREE. The mRNA expression patterns of CsTre1 and CsTre2 in different developmental stages and tissues of C. sinensis were detected by using real-time quantitative PCR (RT-qPCR).[Result] The open reading frame of CsTre1 was 1 701 bp, encoding 566 amino acids, and the protein molecular weight was 64.53 kD. The open reading frame length of CsTre2 was 1 821 bp, encoding 606 amino acids, and the protein molecular weight was 69.08 kD. Signal peptide prediction analysis showed that both front-ends of CsTre1 and CsTre2 had a signal peptide, with positions of 1-16 and 1-17, respectively. The analysis on the secondary structure of the sequence showed that both CsTre1 and CsTre2 were mainly composed of α-helix and random coil, CsTre1 had 24 Sers, 15 Tyrs, and 10 Thrs that may serve as binding sites for protein kinases, while CsTre2 had 27 Sers, 10 Tyrs, and 13 Thrs that may serve as binding sites for protein kinases. The RT-qPCR results revealed that CsTre was expressed throughout all developmental stages of C. sinensis. In the expression pattern of adult stage, the expression level of CsTre1 was much higher than that of CsTre2, and the expression level of male adults of CsTre1 dropped sharply after the fourth day.[Conclusion] The study successfully cloned two trehalose genes of Conopomorpha sinensis Bradley. According to the results of analysis on their molecular characteristics and expression patterns, CsTre1 may be the main gene regulating trehalose metabolism in Conopomorpha sinensis Bradley. The research results provide important clues for elucidating the function of trehalase genes, which lay a solid foundation for the development pest control strategies. [ABSTRACT FROM AUTHOR]

Praying mantids (Mantodea: Mantidae) are iconic insects that have captivated biologists for decades, especially the species with cannibalistic copulatory behavior. This behavior has been cited as evidence that insects lack nociceptive capacities and cannot feel pain; however, this behaviorally driven hypothesis has never been rigorously tested at the genetic or functional level. To enable future studies of nociceptive capabilities in mantids, we sequenced and assembled a draft genome of the Chinese praying mantis (Tenodera sinensis) and identified multiple classes of nociceptive ion channels by comparison to orthologous gene families in Arthropoda. Our assembly—produced using PacBio HiFi reads—is fragmented (total size = 3.03 Gb; N50 = 1.8 Mb; 4,966 contigs), but is highly complete with respect to gene content (BUSCO complete = 98.7% [odb10_insecta]). The size of our assembly is substantially larger than that of most other insects, but is consistent with the size of other mantid genomes. We found that most families of nociceptive ion channels are present in the T. sinensis genome; that they are most closely related to those found in the damp-wood termite (Zootermopsis nevadensis); and that some families have expanded in T. sinensis while others have contracted relative to nearby lineages. Our findings suggest that mantids are likely to possess nociceptive capabilities and provide a foundation for future experimentation regarding ion channel functions and their consequences for insect behavior. [ABSTRACT FROM AUTHOR]

Huanglongbing (HLB) is a severe citrus disease worldwide. Wild Australian limes like Citrus australasica, C. inodora, and C. glauca possess beneficial HLB resistance traits. Individual trees of the three taxa were extensively used in a breeding program for over a decade to introgress resistance traits into commercial-quality citrus germplasm. We generated high-quality, phased, de novo genome assemblies of the three Australian limes using PacBio long-read sequencing. The genome assembly sizes of the primary and alternate haplotypes were determined for C. australasica (337 Mb/335 Mb), C. inodora (304 Mb/299 Mb), and C. glauca (376 Mb/379 Mb). The nine chromosome-scale scaffolds included 86–91% of the genome sequences generated. The integrity and completeness of the assembled genomes were estimated to be at 97.2–98.8%. Gene annotation studies identified 25,461 genes in C. australasica, 27,665 in C. inodora, and 30,067 in C. glauca. Genes belonging to 118 orthogroups were specific to Australian lime genomes compared to other citrus genomes analyzed. Significantly fewer canonical resistance (R) genes were found in C. inodora and C. glauca (319 and 449, respectively) compared to C. australasica (576), C. clementina (579), and C. sinensis (651). Similar patterns were observed for other gene families associated with potential HLB resistance, including Phloem protein 2 (PP2) and Callose synthase (CalS) genes predicted in the Australian lime genomes. The genomic information on Australian limes developed in the present study will help understand the genetic basis of HLB resistance. [ABSTRACT FROM AUTHOR]

Cordyceps militaris is a valuable medicinal fungus which has been widely used as a traditional medicine in East Asia. Compared to the well-known medicinal fungus C. sinensis, C. militaris can produce similar fermented metabolites with various biological activities, but it requires a shorter culture time and simpler culture conditions, and therefore, it has attracted increasing attention in recent years. The purpose of this review was to organize the current studies regarding metabolite production from C. militaris relative to their biological functions. We combined findings of metabolite production to correlate with different fermentation modes to obtain a full view of production processes used to yield the product. While research on C. militaris fermentation is not uncommon to date, its high value still highlights the importance of developing more modern fermentation processes for industrial production. [ABSTRACT FROM AUTHOR]

Reactive oxygen species (ROS) are pivotal in signal transduction processes in plant–pathogen interactions. The ROS signaling pathways involved in Candidatus Liberibacter asiaticus (CLas) and Xanthomonas citri subspecies citri (Xcc) infections in Citrus sinensis (sweet orange) are unclear. In this study, we comprehensively identified ROS metabolism-associated genes, including 9 NADPH oxidase (RBOH), 14 superoxide dismutase (SOD), 1 catalase (CAT), 9 peroxiredoxin (PrxR), 5 ascorbate peroxidase (APX), 4 glutathione peroxidase (GPX), 3 monodehydroascorbate reductase (MDAR), 2 dehydroascorbate reductase (DHAR), 2 glutathione reductase (GR), 24 thioredoxin (Trx), and 18 glutaredoxin (GLR) genes in C. sinensis. An analysis revealed variable gene structures but conserved motifs and domains in ROS subfamilies. A comparative synteny analysis with Arabidopsis thaliana and Vitis vinifera indicated evolutionary conservation of most ROS metabolism-associated genes, with some originating from gene duplication events post-species divergence in C. sinensis. Expression profiling revealed five up-regulated genes and four down-regulated genes during both CLas and Xcc infections. Promoter analysis revealed numerous stress-responsive elements in the promoter of ROS metabolism-associated genes. Protein–protein interaction network analysis highlighted the involvement of ROS metabolism in various biological processes. A comparison of ROS metabolism-associated genes between C. sinensis and Poncirus trifoliata indicated multiple gene gain and loss events within ROS subfamilies of C. sinensis. This study enhances our understanding of ROS metabolism in C. sinensis and sheds light on citrus–pathogen interactions. [ABSTRACT FROM AUTHOR]

Essential oils (EOs) from Citrus sinensis (Rutaceae) possess diverse biological activities. However, a comprehensive comparison of their chemical composition and bioactivity across different plant parts has not been studied yet. The current research comparatively assesses the yield, chemical composition, chiral distribution, antioxidant properties, and larvicidal activity of EOs extracted from the peels, leaves, and flowers of C. sinensis. EOs extracted via hydro-distillation (HD) and steam distillation (SD) were analyzed by gas chromatography–mass spectrometry (GC-MS) and chiral GC-MS to explore their chemical composition and enantiomeric distribution. In addition, their larvicidal and antioxidant potentials were evaluated following standard protocols. Peels of C. sinensis exhibited significantly higher oil content (1.75–2.25%) compared to its leaves (0.75–0.78%) and flowers (0.20–0.25%). The GC-MS analysis identified around 60 compounds, including terpenoids, sesquiterpenoids, and oxygenated terpenoids in the HD and SD extractions. Higher concentrations of sabinene were found in flower extract (38.05–39.89%) and leaf extract (32.30–36.91%), while peel extract contained more than 90% limonene. The larvicidal activity of peel oil was primarily attributed to limonene, with an LC50 value of 0.0031 µL/mL. The current study reports the first chiral (GC-MS) analysis in the essential oil of the leaves and flowers of C. sinensis, paving the way for authenticity and purity. Furthermore, the chemical profiling of citrus EOs, particularly from the peel, demonstrates a safe and promising candidate for diverse biological applications. [ABSTRACT FROM AUTHOR]

The clam Cyclina sinensis is an economically important marine species in China. However, overfishing, habitat destruction, and inbreeding have led to the destruction of genetic resources. In this study, five natural populations [Dandong (DD), Dongying (DY), Tianjin (TJ), Wenzhou (WZ), and Yancheng (YC)] from different geographical locations were relocated and cultured homogeneously to study their potential for translocation and environmental adaptation by observing growth performance and nutrient content. There were significant differences in the growth rates of shell length (SL), shell height (SH), and shell width (SW) among the five populations (P<0.05). The DD, TJ, and YC populations exhibited the highest SL and SH growth rates, whereas these were lowest in the WZ population. The survival rate was highest in the YC population and lowest in the WZ population, which was significantly lower than the other four populations (P<0.05). The DD population exhibited superior plumpness and glycogen content, and the overall glycogen content in male C. sinensis was higher compared with that in females (P<0.05). All populations displayed a high total amino acid content and the essential amino acid/Total Amino Acid (EAA/TAA) and Non-essential amino acid/Total Amino Acid (NEAA/TAA) ratios in the soft tissue of all five populations aligned with the FAO/WHO ideal protein evaluation standards. Despite the same aquaculture environment, however, the nutrient composition of C. sinensis sampled from different populations varied significantly (P<0.05). Therefore, ex situ conservation did not eliminate nutritional differences between the different populations of C. sinensis. These findings highlight the importance of considering environmental and ecological factors in the nutritional assessment and cultivation of shellfish. In addition, C. sinensis from sampling sites closer to the transplantation sites had higher survival rates and growth rates, and the gonadal development of all populations showed adaption to the local environment, resulting in synchronization of the reproductive period. [ABSTRACT FROM AUTHOR]

Background: Citrus is a major fruit crop, and RNA-sequencing (RNA-seq) data can be utilized to investigate its gene functions, heredity, evolution, development, and the detection of genes linked to essential traits or resistance to pathogens. However, it is challenging to use the public RNA-seq datasets for researchers without bioinformatics training, and expertise. Results: OrangeExpDB is a web-based database that integrates transcriptome data of various Citrus spp., including C. limon (L.) Burm., C. maxima (Burm.) Merr., C. reticulata Blanco, C. sinensis (L.) Osbeck, and Poncirus trifoliata (L.) Raf., downloaded from the NCBI SRA database. It features a blast tool for browsing and searching, enabling quick download of expression matrices for different transcriptome samples. Expression of genes of interest can be easily generated by searching gene IDs or sequence similarity. Expression data in text format can be downloaded and presented as a heatmap, with additional sample information provided at the bottom of the webpage. Conclusions: Researchers can utilize OrangeExpDB to facilitate functional genomic analysis and identify key candidate genes, leveraging publicly available citrus RNA-seq datasets. OrangeExpDB can be accessed at http://www.orangeexpdb.com/. [ABSTRACT FROM AUTHOR]

Introduction: Aeromonas hydrophila, a bacterium widely distributed in the natural environment, causes multiple diseases in various animals. Exploring the mechanism of the host defense against A. hydrophila can help develop efficient strategies against Aeromonas infection. Methods: Herein, we investigated the temporal influence of A. hydrophila on the Chinese soft-shelled turtle, an economically important species, at the biochemical, transcriptomic, and metabolomic levels. Plasma parameters were detected with the test kits. Transcriptome and metabolome were respectively applied to screen the differentially expressed genes and metabolites. Results: The contents or activities of these plasma parameters were significantly increased at 24 hpi and declined at 96 hpi, indicating that 24 and 96 hpi were two important time points during infection. Totals of 3121 and 274 differentially expressed genes (DEGs) from the transcriptome while 74 and 91 differentially abundant metabolites (DAMs) from the metabolome were detected at 24 and 96 hpi. The top DEGs at 24 hpi included Ccl2, Ccl3, Ccl4, Il1β, Il6, Il7, Il15, Tnf, and Tnfr1 while Zap70, Cd3g, Cd8a, Itk, Pik3r3, Cd247, Malt1, and Cd4 were the most abundant at 96 hpi. The predominant DAMs included O-phospho-L-serine, γ-Aminobutyric acid, orotate, L-tyrosine, and L-tryptophan at 24 hpi, as well as Lglutamic acid, L-arginine, glutathione, glutathione disulfide, and citric acid at 96 hpi. Discussion: The combined analysis of DEGs and DAMs revealed that tryptophan metabolism, nicotinate and nicotinamide metabolism, as well as starch and sucrose metabolism, were the most important signaling pathways at the early infective stage while tyrosine metabolism, pyrimidine metabolism, as well as alanine, aspartate and glutamate metabolism were the most crucial pathways at the later stage. In general, our results indicated that the Chinese softshelled turtle displays stage-specific physiological responses to resist A. hydrophila infection. [ABSTRACT FROM AUTHOR]

It is well known that application of exogenous trehalose can enhance the heat resistance of plants. To investigate the underlying molecular mechanisms by which exogenous trehalose induces heat resistance in C. sinensis, a combination of physiological and transcriptome analyses was conducted. The findings revealed a significant increase in the activity of superoxide dismutase (SOD) and peroxidase (POD) upon treatment with 5.0 mM trehalose at different time points. Moreover, the contents of proline (PRO), endogenous trehalose, and soluble sugar exhibited a significant increase, while malondialdehyde (MDA) content decreased following treatment with 5.0 mM trehalose under 24 h high-temperature stress (38 °C/29 °C, 12 h/12 h). RNA-seq analysis demonstrated that the differentially expressed genes (DEGs) were significantly enriched in the MAPK pathway, plant hormone signal transduction, phenylpropanoid biosynthesis, flavone and flavonol biosynthesis, flavonoid biosynthesis, and the galactose metabolism pathway. The capability to scavenge free radicals was enhanced, and the expression of a heat shock factor gene (HSFB2B) and two heat shock protein genes (HSP18.1 and HSP26.5) were upregulated in the tea plant. Consequently, it was concluded that exogenous trehalose contributes to alleviating heat stress in C. sinensis. Furthermore, it regulates the expression of genes involved in diverse pathways crucial for C. sinensis under heat-stress conditions. These findings provide novel insights into the molecular mechanisms underlying the alleviation of heat stress in C. sinensis with trehalose. [ABSTRACT FROM AUTHOR]

Background: Opisthorchiasis and clonorchiasis, caused by Opisthorchis viverrini and Clonorchis sinensis, respectively, are significant yet neglected foodborne trematodiases in the Great Mekong Subregion (GMS). Despite the reporting of the prevalence of these human liver flukes in the region over the past decades, there has been a lack of a comprehensive and systematic consolidation of this data. Therefore, we aimed to conduct a thorough systematic review and meta-analysis to synthesize and analyze time-trend prevalence estimates of both O. viverrini and C. sinensis across the GMS for the past 30 years. Methods: This study undertakes a systematic review using a comprehensive search for published articles in PubMed, EMBASE, Scopus, Cochrane and Thai Journal Online databases until early 2023. The pooled prevalence of O. viverrini and C. sinensis infection was analyzed through a random-effects meta-analysis, with meta-regression analysis used to quantify associations with study characteristics. Sub-group analysis was conducted, whenever comparison data were available, to assess the risk of O. viverrini and C. sinensis infection in each GMS country. Heterogeneity among studies was assessed using the Q statistic and quantified by using the I2 Index. Results: From a total of 2997 articles, 155 articles comprising 218 datasets and 751,108 participants were included for review. The GMS prevalence of O. viverrini was 21.11% [45,083/260,237; 95% confidence interval (CI): 17.74–24.47%]. Pooled prevalence estimates were highly observed in Laos (34.06%, 95% CI: 26.85–41.26%), followed by Thailand (18.19%, 95% CI: 13.86–22.51%), and Cambodia (10.48%, 95% CI: 5.52–15.45%). Myanmar and Vietnam had limited data sources for calculation. Clonorchis sinensis infection in GMS was 25.33% (95% CI: 18.32–32.34%), with Guangxi, China, exhibiting the highest prevalence rates at 26.89% (95% CI: 18.34–35.43%), while Vietnam had a prevalence rate of 20.30% (95% CI: 9.13–31.47%). O. viverrini prevalence decreased significantly over time, whereas C. sinensis infection appeared to be stable consistently over time in both China and Vietnam. Conclusions: This comprehensive study, drawing from the largest datasets to date, offers an in-depth systematic prevalence review of human liver flukes in the Greater Mekong Subregion. It underscores the imperative for systematic surveillance, data collection, and the implementation of intervention and control measures for these infectious diseases of poverty. [ABSTRACT FROM AUTHOR]

It is of great significance to explore the molecular mechanism of gametophytic sexual reproduction in the genetic improvement and breeding of tea plants [Camellia sinensis (L.) O. Kuntze]. Imported beta family members (IMBs) are a class of widely distributed nucleoplasmic transport receptor proteins in eukaryotes, affecting plant development and reproduction, and participating in flowering time and sexual reproduction. Still missing, though, is a thorough examination of IMB members in tea plants. Here, seven members of the IMB gene family were screened by a genome-wide investigation in tea plants. These members were scattered unevenly throughout five chromosomes. All of them contained the conserved KAP95 and HEAT Repeat domains. Additionally, the promoter regions of CsIMBs harbored cis-acting elements associated with plant hormones, light, and abiotic stress responses. In order to further confirm the function of CsIMBs in the sexual reproduction of tea plants, the expression patterns of CsIMBs in different flower development stages and the ovary (before and after pollination) were analyzed. The expression results highlighted that CsIMBs were related to the fertility and fruiting of tea plants. Furthermore, five of the seven CsIMBs (CsIMB1a, CsIMB1b, CsIMB2, CsIMB3a, and CsIMB3b) were found to be localized in the nucleus revealed by subcellular localization analysis. These results offer a comprehensive characterization of IMB genes as well as insights into the potential roles of CsIMBs participating in the gametophytic sexual reproduction of C. sinensis. [ABSTRACT FROM AUTHOR]

Cordyceps sinensis is a precious medicinal and edible fungus, which is widely used in body health care and disease prevention. The current research focuses on the comparison of metabolite characteristics between a small number of samples and lacks a comprehensive evaluation of the quality of C. sinensis in a large-scale space. In this study, LC-MS/MS, principal component analysis (PCA), hierarchical cluster analysis (HCA), and the membership function method were used to comprehensively evaluate the characteristics and quality of metabolites in 15 main producing areas of C. sinensis in China. The results showed that a total of 130 categories, 14 supercategories, and 1718 metabolites were identified. Carboxylic acids and derivatives, fatty acyls, organo-oxygen compounds, benzene and substituted derivatives, prenol lipids, and glycerophospholipids were the main components of C. sinensis. The HCA analysis and KEGG pathway enrichment analysis of 559 differentially accumulated metabolites (DAMs) showed that the accumulation models of fatty acids and conjugates and carbohydrates and carbohydrate conjugates in glycerophospholipid metabolism and arginine and proline metabolism may be one of the reasons for the quality differences in C. sinensis in different producing areas. In addition, a total of 18 biomarkers were identified and validated, which had a significant discrimination effect on the samples (p < 0.05). Overall, YS, BR, and ZD, with the highest membership function values, are rich and balanced in nutrients. They are excellent raw materials for the development of functional foods and provide scientific guidance for consumers to nourish health care. [ABSTRACT FROM AUTHOR]

Background: The purpose of the present study was to enhance the memory-boosting activity of the standardized hydroalcoholic Camellia sinensis extract (CSE) by the formation of nanophytosomes with Leciva S70 phospholipid. The central composite design was used to optimize the solvent evaporation method for the formulation of C. sinesis phytosomes (CSP). Results: The optimized formulation had a mean particle size of 212.3 nm ± 0.39, PDI of 0.238 ± 0.0197, and zeta potential of −42.02 ± 0.995 mV. C. sinensis phytosome formation was confirmed by analytical techniques. The aqueous solubility of the developed CSP was 95.92 ± 0.31, which is 7.34 times greater than that of pure CSE (13.07 ± 0.19). CSP was found more effective than either pure CSE (26.42 ± 0.4654%) or the physical mixture (32.15 ± 0.4596%) in releasing the CSE from the formulation (72.16 ± 0.5248%). Acute toxicity study corroborated the safety of CSP in rats. CSP demonstrated a significant (p < 0.05) reduction in escape and transferred latency on both days (15th and 16th) as compared to CSE, indicating the improvement of the memory-boosting activity. Furthermore, CSP-treated rats significantly improved acetylcholine (Ach) levels and brain tissue concentration compared with CSE. Moreover, the phytosomal formulation of CSP exhibited its rationality with an improvement of bioavailability by 3.21 folds compared with pure CSE. Conclusion: The presence of phospholipids in the CSP formulation and the formation of smaller particles may aid in crossing the blood–brain barrier, increasing brain tissue concentration and bioavailability. This, in turn, leads to an increase in memory-boosting activity. [ABSTRACT FROM AUTHOR]

Background: Trichomonas gallinae is a parasite that causes canker and severe loss and death, especially in young pigeons. Metronidazole (MTZ) is the recommended drug for treating avian trichomoniasis. Due to drug resistance, non‐chemical alternatives, such as medicinal plant extracts, are also considered possible therapies for this disease. Objectives: This study compares the antitrichomonal effects of MTZ with extracts of Camellia sinensis and Ziziphus vulgaris on T. gallinae in vitro. Methods: Samples of T. gallinae were taken from infected pigeons. Multi‐well plates with different concentrations (5, 10, 25, 50 and 100 µg/mL) of plant extracts were used for the in vitro study. Results: The minimum inhibitory concentration (MIC) of C. sinensis extract was 25 µg/mL over 24 h, compared to 50 µg/mL for MTZ. The MIC value of the Z. vulgaris extracts was 50 µg/mL. Conclusions: The results suggest that the extracts of Z. vulgaris and C. sinensis, as potential natural agents, could have anti‐avian trichomoniasis properties. This study also shows that MTZ, C. sinensis and Z. vulgaris are equally effective in preventing the growth of T. gallinae trophozoites in the culture. [ABSTRACT FROM AUTHOR]

Sigma-class glutathione transferase (GST) proteins with dual GST and prostaglandin synthase (PGS) activities play a crucial role in the establishment of Clonorchis sinensis infection. Herein, we analyzed the structural and enzymatic properties of sigma-class GST (CsGST-σ) proteins to obtain insight into their antioxidant and immunomodulatory functions in comparison with mu-class GST (CsGST-μ) proteins. CsGST-σ proteins conserved characteristic structures, which had been described in mammalian hematopoietic prostaglandin D2 synthases. Recombinant forms of these CsGST-σ and CsGST-μ proteins expressed in Escherichia coli exhibited considerable degrees of GST and PGS activities with substantially different specific activities. All recombinant proteins displayed higher affinities toward prostaglandin H2 (PGS substrate; average Km of 30.7 and 3.0 μm for prostaglandin D2 [PGDS] and E2 synthase [PGES], respectively) than those toward CDNB (GST substrate; average Km of 1,205.1 μm). Furthermore, the catalytic efficiency (Kcat/Km) of the PGDS/PGES activity was higher than that of GST activity (average Kcat/Km of 3.1, 0.7, and 7.0×10-3 s-1μm-1 for PGDS, PGES, and GST, respectively). Our data strongly suggest that the C. sinensis sigma- and mu-class GST proteins are deeply involved in regulating host immune responses by generating PGD2 and PGE2 in addition to their roles in general detoxification. [ABSTRACT FROM AUTHOR]

Purpose: Garcinia quaesita is an exquisite spice used extensively in imparting flavor to foods, and in curing diseases in Ayurveda. Considering the seasonality of the fruit rind, abundance of the leaves, and potential functional properties displayed by the leaves, this study was intended to develop functional foods incorporating garcinia leaf extract and assess their sensory, functional, and nutritional properties. Research Method: Herbal tea, herbal crackers, herbal appetizers and herbal confectionery incorporated with garcinia leaves were developed and assessed for their sensory qualities. As in functional properties, total polyphenol content, total antioxidant activity, and total flavonoid content of the products were measured while the proximate composition was analyzed to obtain basic nutritional composition. Findings: All the products were sensory accepted; with mean scores above, "3" from a 5-point hedonic scale in each attribute, while retaining the basic nutritional composition of their particular food categories. Each product reported significant amounts of polyphenols and flavonoids with remarkable antioxidant activity, whereas, the highest reported values were shown by Herbal tea, consisting of G. quaesita, P. guajava, C. verum and C. sinensis. Being comprised of direct herbal infusions as well as the synergetic effect of the herbal materials seems the potential factors behind these elevated functional properties. Research Limitations: The main research limitation was the limited accessibility to sensory panelists, raw materials, chemicals and laboratories due to the pandemic situation during the study period. Originality: The study demonstrates the concept of incorporating Garcinia quaesita leaf extract in food products to impart functional properties while retaining higher sensory acceptability. [ABSTRACT FROM AUTHOR]

Enterohemorrhagic E. coli (EHEC) is a group of pathogenic bacteria that is associated with worldwide human foodborne diarrheal illnesses and the development of hemolytic uremic syndrome, a potentially deadly condition associated with Shiga toxins (Stxs). Currently, approved vaccines for human prophylaxis against infection do not exist, and one barrier preventing the successful creation of EHEC vaccines is the absence of dependable animal models, including mice, which are naturally resistant to EHEC infection and do not manifest the characteristic signs of the illness. Our lab previously developed gold nanoparticle (AuNP)-based EHEC vaccines, and assessed their efficacy using Citrobacter rodentium, which is the mouse pathogen counterpart of EHEC, along with an Stx2d-producing strain that leads to more consistent disease kinetics in mice, including lethality. The purpose of this study was to continue evaluating these vaccines to increase protection. Here, we demonstrated that subcutaneous immunization of mice with AuNPs linked to the EHEC antigens EscC and intimin (Eae), either alone or simultaneously, elicits functional robust systemic humoral responses. Additionally, vaccination with both antigens together showed some efficacy against Stx2d-producing C. rodentium while AuNP-EscC successfully limited infection with non-Stx2d-producing C. rodentium. Overall, the collected results indicate that our AuNP vaccines have promising potential for preventing disease with EHEC, and that evaluation of novel vaccines using an appropriate animal model, like C. rodentium described here, could be the key to finally developing an effective EHEC vaccine that can progress into human clinical trials. [ABSTRACT FROM AUTHOR]

The liver fluke disease caused by Clonorchis sinensis is one of the most serious food-borne parasitic diseases in China. Many freshwater fish and shrimps can be infected with C. sinensis metacercariae as the second intermediate hosts in endemic regions. Owing to the lack of infected humans and the good administration of pet dogs and cats in cities of non-endemic regions, few fish are expected to be infected with C. sinensis metacercariae in urban lakes. To determine the infection of C. sinensis metacercariae in freshwater fish and shrimps in urban lakes, a total of 18 fish species and one shrimp species were investigated in the East Lake of Wuhan City. Metacercariae were isolated by artificial digestive juice and identified using morphology and rDNA-ITS2 sequences. Five species of fish, Pseudorasbora parva, Ctenogobius giurinus, Squalidus argentatus, Hemiculter leuciclus, and Rhodeus spp., were infected with C. sinensis metacercariae. The overall prevalence of C. sinensis was 32.5%. The highest prevalence was found in P. parva with 57.9%, while S. argentatus exhibited the highest mean abundance (13.9). Apart from the C. sinensis metacercariae, four species of other trematode metacercariae were also identified across twelve fish species in total. Owing to the consumption of undercooked fish and feeding cats with small fish caught by anglers, there is a potential risk that the small fish infected with C. sinensis metacercariae may act as an infection source to spread liver fluke. Given the complete life cycle of C. sinensis, stray cats and rats were inferred to act as the important final hosts of C. sinensis in urban lakes in non-endemic areas. [ABSTRACT FROM AUTHOR]

Clonorchis sinensis is one of the most important fish-borne zoonotic parasitic worms in humans, and is distributed in several countries with more than 15 million people infected globally. However, the lack of a point-of-care testing (POCT) method is still the critical barrier to effectively prevent clonorchiasis. With the application of novel fluorescent nanomaterials, the development of on-site testing methods with high signal enhancement can provide a simple, precise and inexpensive tool for disease detection. In this study, Eu-(III) nanoparticles (EuNPs) were used as indicative probes, combined with C. sinensis tandem repeat sequence 1 (CSTR1) antigen to capture specific antibodies. Afterward, the complex binds to mouse anti-human IgG immobilized on the test line (T-line) producing a fluorescent signal under UV light. The EuNPs-fluorescent immunoassay (EuNPs-FIA) was successfully constructed, allowing sample detection within 10 min. It enabled both qualitative determination with the naked eye under UV light and quantitative detection by scanning the fluorescence intensity on the test line and control line (C-line). A total of 133 clinical human sera (74 negative, 59 clonorchiasis, confirmed by conventional Kato-Katz (KK) methods and PCR via testing fecal samples corresponding to each serum sample) were used in this study. For qualitative analysis, the cut-off value of fluorescence for positive serum was 31.57 by testing 74 known negative human samples. The assay had no cross-reaction with other 9 parasite-infected sera, and could recognize the mixed infection sera of C. sinensis and other parasites. The sensitivity and specificity of EuNPs-FIA were both 100% compared with KK smear method. Taking advantage of its high precision and user-friendly procedure, the established EuNPs-FIA provides a powerful tool for the diagnosis and epidemiological survey of clonorchiasis. Author summary: Clonorchis sinensis is one of the most important foodborne zoonotic parasites, affecting over 15 million people worldwide. However, the low-intensity infection and the lack of instant detection methods remain the main inhibitory factors for controlling this disease. In this study, the specific CSTR1 antigen was combined with EuNPs to construct a fluorescent probe to capture clonorchiasis-specific antibodies. Then, the complex binds to anti-human IgG immobilized on the test line, producing a fluorescent signal under UV light. The established EuNPs-FIA method enabled both qualitative determination with the naked eye and quantitative detection by scanning fluorescence probes intensity. The FIA has no cross-reaction with other 9 heterologous samples, and the sensitivity tests showed that the serum sample corresponding to 24 eggs per gram (EPG) in KK method can be still detected. Our work indicated that the established EuNPs-FIA method has broad application prospects in clinical and epidemiological studies and can aid in the surveillance and control of clonorchiasis. [ABSTRACT FROM AUTHOR]

Citrus canker disease affects citrus production. This disease is caused by Xanthomonas citri subsp. citri (Xcc). Previous studies confirmed that during Xcc infection, PthA4, a transcriptional activator like effector (TALE), is translocated from the pathogen to host plant cells. PthA4 binds to the effector binding elements (EBEs) in the promoter region of canker susceptibility gene LOB1 (EBEPthA4-LOBP) to activate its expression and subsequently cause canker symptoms. Previously, the Cas12a/CBE co-editing method was employed to disrupt EBEPthA4-LOBP of pummelo, which is highly homozygous. However, most commercial citrus cultivars are heterozygous hybrids and more difficult to generate homozygous/biallelic mutants. Here, we employed Cas12a/CBE co-editing method to edit EBEPthA4-LOBP of Hamlin (Citrus sinensis), a commercial heterozygous hybrid citrus cultivar grown worldwide. Binary vector GFPp1380N-ttLbCas12a:LOBP1-mPBE:ALS2:ALS1 was constructed and shown to be functional via Xcc-facilitated agroinfiltration in Hamlin leaves. This construct allows the selection of transgene-free regenerants via GFP, edits ALS to generate chlorsulfuron-resistant regenerants as a selection marker for genome editing resulting from transient expression of the T-DNA via nCas9-mPBE:ALS2:ALS1, and edits gene(s) of interest (i.e., EBEPthA4-LOBP in this study) through ttLbCas12a, thus creating transgene-free citrus. Totally, 77 plantlets were produced. Among them, 8 plantlets were transgenic plants (#HamGFP1 - #HamGFP8), 4 plantlets were transgene-free (#HamNoGFP1 - #HamNoGFP4), and the rest were wild type. Among 4 transgene-free plantlets, three lines (#HamNoGFP1, #HamNoGFP2 and #HamNoGFP3) contained biallelic mutations in EBEpthA4, and one line (#HamNoGFP 4) had homozygous mutations in EBEpthA4. We achieved 5.2% transgene-free homozygous/biallelic mutation efficiency for EBEPthA4-LOBP in C. sinensis cv. Hamlin, compared to 1.9% mutation efficiency for pummelo in a previous study. Importantly, the four transgenefree plantlets and 3 transgenic plantlets that survived were resistant against citrus canker. Taken together, Cas12a/CBE co-editing method has been successfully used to generate transgene-free canker-resistant C. sinensis cv. Hamlin in the T0 generation via biallelic/homozygous editing of EBEpthA4 of the canker susceptibility gene LOB1. [ABSTRACT FROM AUTHOR]

The sweet oranges [C. sinensis (L.) Osbeck] produced in India are mainly consumed fresh as a table fruit and in the form of freshly extracted juice. Currently, a fraction of the fruit is processed into products like orange juice, concentrates, pulp, and other value-added products. Seedless varieties are preferred both by the consumers and by the processing industry; however, indigenously developed seedless sweet orange cultivars are not available. Citrus triploids are usually seedless due to their abnormal meiosis and embryo abortion. A research study was undertaken at CCRI Nagpur to develop triploid seedless plants of the local sweet orange cultivar Mosambi through endosperm culture in the shortest possible time by dovetailing various techniques. Various steps, viz., endosperm excision, standardization of media for callus initiation, somatic embryogenesis, shoot/root differentiation, mini-grafting, and validation of the triploid status through flow cytometry, chromosome counting, and other morphological parameters, were standardized, and seven confirmed triploid plants were finally produced. An evaluation of fruit quality parameters during the 2022 and 2023 fruiting seasons revealed that the fruits of the triploid sweet orange trees were commercially seedless. This is the first reported comprehensive study on the successful development of commercially seedless plants of the sweet orange cultivar Mosambi. The fruits of the triploid plants showed desirable fruit quality parameters in terms of their seed number (3 to 5.9/fruit), higher vitamin C content (34.4 to 42.66 mg), and lower limonene content (7.77 to 11.34 µg/mL). These triploid plants have the potential to gain recognition as a distinct variety. [ABSTRACT FROM AUTHOR]

Objective: Foodborne zoonotic trematodes (FZT), which infect the liver, lungs and intestines of humans, are an emerging public health concern in tropical countries including Vietnam. In northern Vietnam, Clonorchis sinensis is recognised as the most important species of the FZT. Up to now, small‐scale studies conducted in the country have indicated that health education could improve participants' knowledge and practices related to clonorchiasis, however strong evidence is still lacking. We conducted an intervention trial between 2020 and 2021 in four communes in Yen Bai and Thanh Hoa province, aiming to evaluate the impact of an extended educational package on knowledge, attitude and practices related to clonorchiasis, as well as on knowledge on the large liver fluke and minute intestinal flukes. Methods: To this end, baseline and post‐intervention questionnaires were conducted. Generalised estimating equations models were run to analyse the impact of the intervention on knowledge, attitudes and practices over time in the intervention versus control group. Finally, 172 people in the intervention group and 162 in the control group were included for analysis. Results: Results demonstrated that the intervention significantly improved clonorchiasis knowledge, attitudes and practices, with higher odds for a correct knowledge response (odds ratio (OR) = 2.80, 95% confidence interval (CI) = 1.84–4.27, p‐value < 0.001), increased average mean attitude score (mean = 0.363, 95%CI = 0.182–0.544, p‐value < 0.001), and a reduced odds of consuming raw fish (OR = 0.15, 95%CI = 0.06–0.40, p = 0.002) in the intervention group compared to the control group post‐intervention versus baseline. Additionally, participants in the intervention group exhibited enhanced basic knowledge of the large liver fluke and minute intestinal flukes post‐intervention. Conclusions: These findings demonstrate the potential of health education programs in preventing clonorchiasis in endemic areas, emphasising the importance of continued health education as a critical component of integrated control programs for clonorchiasis. [ABSTRACT FROM AUTHOR]