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32 pages, 39042 KB  
Article
Molecular Phylogeny and Species Delimiting for the Genus Hoplolaimus (Nematoda: Tylenchida) with Description of Hoplolaimus floridensis sp. n. and Notes on Biogeography of the Genus in the United States
by Sergei A. Subbotin, Mihail Kantor, Erika Consoli, Niclas H. Lyndby, Amy Michaud, Zafar Handoo and Renato N. Inserra
Int. J. Mol. Sci. 2025, 26(17), 8501; https://doi.org/10.3390/ijms26178501 (registering DOI) - 1 Sep 2025
Abstract
Lance nematodes, Hoplolaimus spp., feed on the roots of many kinds of plants, including agronomic crops. In this study, morphological and molecular analyses of several Hoplolaimus species and populations are provided. We were able to collect and characterize the topotype materials of H. [...] Read more.
Lance nematodes, Hoplolaimus spp., feed on the roots of many kinds of plants, including agronomic crops. In this study, morphological and molecular analyses of several Hoplolaimus species and populations are provided. We were able to collect and characterize the topotype materials of H. galeatus from Arlington, Virginia; H. stephanus syn. n. from Nichols, South Carolina; and H. concaudajuvencus from Pensacola, Florida, and several additional populations and species from the United States, Israel, and India. Phylogenetic analyses of several hundred sequences of the D2–D3 expansion regions of 28S rRNA, ITS rRNA, and COI genes of Hoplolaimus species obtained from published and original datasets were given. Fifty-three new D2–D3 of 28S rRNA, 43 new ITS rRNA, and 47 new COI sequences from 23 isolates of Hoplolaimus spp. and one isolate of Peltamigratus christiei were obtained in this study. New molecular identities for H. concaudajuvencus and H. galeatus were proposed. Hoplolaimus stephanus syn. n. was considered a synonym of H. galeatus based on the morphological and molecular similarity of these two species. Analysis of morphology and molecular data did not reveal significant differences among H. columbus syn. n., H. indicus syn. n., and H. seinhorsti, and the first two species were synonymized with H. seinhorsti. A new species, H. floridensis sp. n., was described from many locations in Florida, USA. It was separated from other representatives of the genus Hoplolaimus by its morphological and molecular characteristics. Maps with geographical distribution of several lance nematode species in North America were reconstructed based on published and original molecular identification of samples. Full article
(This article belongs to the Special Issue Advances in Plant Nematology Research)
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14 pages, 550 KB  
Article
Rumen Microbiota in Cattle and Buffaloes: Insights into Host-Specific Bacterial Diversity
by Shyam Sundar Paul, Avijit Dey, Daoharu Baro, Jerome Andonissamy, Jyotirmoyee Paul and Balbir Singh Punia
Biology 2025, 14(9), 1166; https://doi.org/10.3390/biology14091166 - 1 Sep 2025
Abstract
The present investigation was designed to elucidate the comparative collective diversity of bacteria in the rumen of buffalo and cattle. For this study, a total of 14,913 16S rRNA gene (rrn) sequences generated through Sanger sequencing of ruminal bacteria deposited in the GenBank [...] Read more.
The present investigation was designed to elucidate the comparative collective diversity of bacteria in the rumen of buffalo and cattle. For this study, a total of 14,913 16S rRNA gene (rrn) sequences generated through Sanger sequencing of ruminal bacteria deposited in the GenBank database were analyzed, of which 13,432 sequences were from cattle and 1481 sequences were from buffalo. Bacterial sequences of cattle origin represented 18 existing phyla and 165 genera, and those of buffalo origin represented 12 phyla and 67 genera. According to Ribosomal Database Project (RDP) classifier, Firmicutes was the dominant phylum in cattle, representing 47.9% of all sequences. Bacteroidetes was the second most abundant phylum (32.3% of sequences), while Proteobacteria accounted for 8.6% of total sequences. In buffalo, Firmicutes was the dominant phylum with 47.2% of total sequences. Bacteroidetes and Proteobacteria phyla constituted 38.3% and 4.6% of total sequences, respectively. We identified 172 shared non-rare species-level operational taxonomic units (OTUs) between buffalo and cattle, with 17 unique to buffalo belonging to three phyla: Bacteroidetes, Firmicutes, and Fibrobacteres. In cattle, 774 OTUs of unique sequences were assigned to six phyla, namely, Firmicutes (422 OTUs), Bacteroidetes (234 OTUs), Fibrobacteres (99 OTUs), Actinobacteria (7 OTUs), Cyanobacteria (5 OTUs), and SR1 (7 OTUs). This study revealed significant differences in rumen bacterial diversity between buffaloes and cattle, supporting the development of species-specific strategies to enhance fibrous feed utilization. Full article
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18 pages, 2575 KB  
Article
The First Whole Genome Sequence and Methylation Profile of Gerronema lapidescens QL01
by Yanming Qiao, Zhiyuan Jia, Yuying Liu, Na Zhang, Chun Luo, Lina Meng, Yajie Cheng, Minglei Li, Xiuchao Xie and Jianzhao Qi
J. Fungi 2025, 11(9), 647; https://doi.org/10.3390/jof11090647 (registering DOI) - 1 Sep 2025
Abstract
Gerronema lapidescens (Lei Wan), a valued medicinal basidiomycete traditionally employed for antiparasitic and digestive ailments, faces severe conservation threats due to unsustainable wild harvesting and the absence of reliable cultivation protocols. To address this crisis and unlock its pharmacotherapeutic potential, we present the [...] Read more.
Gerronema lapidescens (Lei Wan), a valued medicinal basidiomycete traditionally employed for antiparasitic and digestive ailments, faces severe conservation threats due to unsustainable wild harvesting and the absence of reliable cultivation protocols. To address this crisis and unlock its pharmacotherapeutic potential, we present the first chromosome-scale genome assembly and comprehensive methylome profile for the wild strain G. lapidescens QL01, domesticated from the Qinling Mountains. A multi-platform sequencing strategy (Illumina and PacBio HiFi) yielded a high-quality 82.23 Mb assembly anchored to 11 chromosomes, exhibiting high completeness (98.4% BUSCO) and 46.03% GC content. Annotation predicted 15,847 protein-coding genes, with 81.12% functionally assigned. Genome-wide analysis identified 8.46 million high-confidence single-nucleotide polymorphisms (SNPs). Notably, methylation profiling revealed 3.25 million methylation events, with elevated densities on chromosomes 4, 9, and 10, suggesting roles in gene silencing and environmental adaptation. Phylogenomic analyses clarified the evolutionary status of G. lapidescens, whilst gene family evolution indicated moderate dynamics reflecting niche adaptation. Carbohydrate-Active enzymes (CAZymes) analysis identified 521 enzymes, including 211 Glycoside Hydrolases (GHs), consistent with organic matter degradation. Additionally, 3279 SSRs were catalogued as molecular markers. This foundational resource elucidates G. lapidescens’s genetic architecture, epigenetic regulation, evolutionary history, and enzymatic toolkit, underpinning future research into medicinal compound biosynthesis, environmental adaptation, germplasm conservation, and sustainable cultivation. Full article
16 pages, 6039 KB  
Article
Blue Light Receptor WC-2 Regulates Ganoderic Acid Biosynthesis in Ganoderma lingzhi
by Yan Xu, Xiong-Min Huang, Zi-Xu Wang, Ying-Jie Zhao, Dong-Mei Lv and Jun-Wei Xu
J. Fungi 2025, 11(9), 646; https://doi.org/10.3390/jof11090646 (registering DOI) - 1 Sep 2025
Abstract
Ganoderic acid (GA) is a key bioactive component with pharmacological properties that is found in Ganoderma lingzhi, a renowned medicinal mushroom. Currently, the regulatory mechanisms underlying GA biosynthesis in G. lingzhi remain to be further elucidated. In this study, blue light induction [...] Read more.
Ganoderic acid (GA) is a key bioactive component with pharmacological properties that is found in Ganoderma lingzhi, a renowned medicinal mushroom. Currently, the regulatory mechanisms underlying GA biosynthesis in G. lingzhi remain to be further elucidated. In this study, blue light induction was found to significantly enhance the GA content in G. lingzhi. To explore the regulatory mechanism of GA biosynthesis in response to blue light, the blue light receptor WC-2 was identified, and its regulatory role was characterized. The deletion of wc-2 resulted in a significant reduction in both GA content and the accumulation of intermediates compared to the wild-type control strain, largely due to the strong downregulation of key GA biosynthetic genes. Additionally, decreased asexual spore production and reduced expression of sporulation-specific genes were observed with the deletion of wc-2. The overexpression of wc-2 led to greatly enhanced GA accumulation. Under blue light induction, the maximum contents of GA-Mk, GA-T, GA-S, and GA-Me were 2.27-, 2.51-, 2.49-, and 2.08-fold higher, respectively, compared to the control kept in darkness. These results demonstrate that the blue light receptor WC-2 functions as a positive regulator of GA biosynthesis in G. lingzhi, influencing the expression of genes involved in GA biosynthesis and asexual spore production, thereby advancing our understanding of the intricate regulatory network of GA biosynthesis. Full article
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17 pages, 1315 KB  
Article
MfWRKY40 Positively Regulates Drought Tolerance in Arabidopsis thaliana by Scavenging Reactive Oxygen Species
by Xueli Zhang, Wei Duan, Yuxiang Wang, Zhihu Jiang and Qian Li
Int. J. Mol. Sci. 2025, 26(17), 8495; https://doi.org/10.3390/ijms26178495 (registering DOI) - 1 Sep 2025
Abstract
Drought stress is a major abiotic constraint that severely restricts the growth of Medicago falcata L. by inducing the accumulation of reactive oxygen species (ROS) in plants. WRKY transcription factors (TFs) play a key role in regulating plant responses to drought stress. In [...] Read more.
Drought stress is a major abiotic constraint that severely restricts the growth of Medicago falcata L. by inducing the accumulation of reactive oxygen species (ROS) in plants. WRKY transcription factors (TFs) play a key role in regulating plant responses to drought stress. In this study, we investigated the role of the MfWRKY40 gene in drought tolerance. Under mannitol and ABA stress treatments, MfWRKY40-overexpressing lines (OEs) showed significantly longer primary roots, increased lateral roots, and higher fresh weight compared to wild-type (Col) lines, indicating significantly enhanced growth and drought tolerance. Similarly, under soil drought conditions, transgenic Arabidopsis thaliana exhibited enhanced drought tolerance. NBT staining demonstrated decreased ROS accumulation in transgenic lines after stress treatment. Correspondingly, the MfWRKY40-overexpressing lines displayed significantly lower levels of hydrogen peroxide (H2O2), superoxide anion (O2), and malondialdehyde (MDA) compared to Col, along with elevated activities of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD), as well as increased proline (Pro) content. Furthermore, MfWRKY40 upregulated the expression of antioxidant enzyme genes (AtPOD3, AtSOD4, and AtCAT1) and modulated the expression of other drought-related genes. In summary, our results demonstrate that MfWRKY40 enhances drought tolerance in A. thaliana by improving ROS scavenging capacity. This study provides a theoretical foundation for further exploration of MfWRKY40’s functional mechanisms in drought stress adaptation. Full article
(This article belongs to the Special Issue Abiotic Stress in Plants: Physiological and Molecular Responses)
37 pages, 2247 KB  
Review
The Central Nervous System Modulatory Activities of N-Acetylcysteine: A Synthesis of Two Decades of Evidence
by Desislava Ivanova Cherneva, Gabriela Kehayova, Simeonka Dimitrova and Stela Dragomanova
Curr. Issues Mol. Biol. 2025, 47(9), 710; https://doi.org/10.3390/cimb47090710 (registering DOI) - 1 Sep 2025
Abstract
N-acetylcysteine (NAC) has garnered increasing interest for its neurotherapeutic capabilities beyond its recognized functions as a mucolytic agent and an antidote for acetaminophen toxicity. This review consolidates findings from both preclinical and clinical studies to investigate NAC’s diverse modulatory effects on the central [...] Read more.
N-acetylcysteine (NAC) has garnered increasing interest for its neurotherapeutic capabilities beyond its recognized functions as a mucolytic agent and an antidote for acetaminophen toxicity. This review consolidates findings from both preclinical and clinical studies to investigate NAC’s diverse modulatory effects on the central nervous system (CNS). NAC primarily functions as an antioxidant by replenishing glutathione and mitigating oxidative stress; however, it produces glutathione-independent effects through the modulation of mitochondrial redox systems, ferroptosis, and the Nrf2-ARE signaling pathway. It plays a significant role in neuroinflammatory processes by inhibiting the production of cytokines, the expression of iNOS, and the activation of microglia. Furthermore, NAC affects various neurotransmitter systems—including glutamatergic, dopaminergic, GABAergic, serotonergic, cholinergic, and adrenergic pathways—by modulating synaptic transmission, receptor activity, and transporter functionality. It promotes neuroprotection through the enhancement of neurotrophic factors, the preservation of mitochondrial integrity, and the upregulation of survival signaling pathways. Recent evidence also emphasizes NAC’s role in gene expression and the regulation of cortisol levels. The extensive range of NAC’s neurobiological effects highlights its therapeutic potential in treating neurodegenerative and neuropsychiatric disorders. Nevertheless, the variability in clinical outcomes indicates a pressing need for more focused, mechanism-based research. Full article
(This article belongs to the Section Molecular Pharmacology)
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16 pages, 7237 KB  
Article
In Silico Analysis of Two Hard Tick P450s: Identification, Characterization, and Putative Metabolism of Cymbopogon citratus Essential Oil Constituents
by Caishan Li, Licui Wen, Wenyu Shi, Yuqian Deng, Na Zhou, Xueqing Zhao, Qingyong Guo and Bayinchahan Gailike
Int. J. Mol. Sci. 2025, 26(17), 8489; https://doi.org/10.3390/ijms26178489 (registering DOI) - 1 Sep 2025
Abstract
The cytochrome p450 gene family is widely involved in various biological processes in arthropods. Tick p450s are often associated with chemical acaricides, but knowledge of their involvement in the metabolism of plant-derived essential oil components is limited. In this study, we identified the [...] Read more.
The cytochrome p450 gene family is widely involved in various biological processes in arthropods. Tick p450s are often associated with chemical acaricides, but knowledge of their involvement in the metabolism of plant-derived essential oil components is limited. In this study, we identified the non-redundant number of p450 transcripts (NRNPTs) from Haemaphysalis longicornis and Hyalomma asiaticum under the Cymbopogon citratus essential oil (CCEO) and terpinolene stress using de novo transcriptome data, respectively. In this study, we identified and characterized the NRNPTs of Ha. longicornis and Hy. asiaticum. Their gene expression patterns and biological functions under CCEO and terpinolene stress were further analyzed. Finally, Hy. asiaticum NRNPTs (87) were more numerous than Ha. longicornis (58). Phylogenetic analyses showed that NRNPTs of both Hy. asiaticum and Ha. longicornis could be categorized in clan 2, clan 3, clan 4, and clan mito, this data comes from the NRNPTs. Phylogenetic analyses showed that NRNPTs of both Hy. asiaticum and Ha. longicornis could be categorized in clan 2, clan 3, clan 4, and clan mito. p450 members of both were most distributed in clan 3. In addition, one Hy. asiaticum NRNPT was identified as belonging to the new classification clan 20 (HyasCYP20A1). The biological functions and pathways of p450 family members enriched in Hy. asiaticum and Ha. longicornis under different exogenous substance stresses were different, and the expression patterns of these genes were inconsistent. Molecular docking results showed that Ha. longicornis p450 members (HaloCYP3A4 and HaloCYP4B1), which were significantly up-regulated under CCEO stress, as well as Hy. asiaticum HyasCYP24A1 and HyasCYP4V2 (the HaloCYP3A4 and HaloCYP4B1 homologous genes), encode proteins that differ in their ability to metabolize CCEO components, but they all bind well to Germacrene D and naphthalene. Our study enriches the knowledge of the involvement of p450 family members of different tick species in the metabolism of essential oil components of plants, and provides a theoretical basis for further in-depth studies on the function of tick p450 enzymes. Full article
(This article belongs to the Section Bioactives and Nutraceuticals)
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23 pages, 2096 KB  
Review
Epigenetic Mechanisms Associated with Livestock Adaptation to Heat Stress
by Sundar Aravindh, Mullakkalparambil Velayudhan Silpa, Santhi Priya Voggu, Ebenezer Binuni Rebez, Gajendirane Kalaignazhal, Mouttou Vivek Srinivas, Frank Rowland Dunshea and Veerasamy Sejian
Biology 2025, 14(9), 1154; https://doi.org/10.3390/biology14091154 - 1 Sep 2025
Abstract
The livestock sector, a crucial source of revenue and global food security, is facing serious challenges due to climate change driven by global warming. This leads to serious effects on animal health and productivity, making it difficult for the livestock industry to meet [...] Read more.
The livestock sector, a crucial source of revenue and global food security, is facing serious challenges due to climate change driven by global warming. This leads to serious effects on animal health and productivity, making it difficult for the livestock industry to meet the global demand and sustain the livelihoods of farmers. The main factor affecting livestock’s productivity is heat stress. However, animals develop various adaptive mechanisms to cope with the effects of heat stress. Cellular and molecular responses act as key defense mechanisms, enabling animals adapt to environmental changes. The recent advancements in molecular biology have opened up opportunities for extensive research on epigenetics, which has a key role in regulating gene expression in animals in response to environmental stimuli. Such studies have gained considerable attention regarding heat acclimation in animals due to the fact that epigenetic mechanisms have been recognized as key players in long-term adaptation to high temperatures in farm animals. This review summarizes the different mechanisms and methodologies used to assess heat stress-associated epigenetic changes, including DNA methylation, which is an extensively studied epigenetic regulatory mechanism in relation to gene expression. The review also highlights the mechanisms and regulation of adaptation to heat stress in animals and collates information related to various epigenetic markers to assess the heat stress response, thereby aiding in improving thermal resilience in animals. Full article
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22 pages, 2681 KB  
Article
Genome Sequence and Characterization of Bacillus cereus Endophytes Isolated from the Alectra sessiliflora and Their Biotechnological Potential
by Khuthadzo Tshishonga and Mahloro Hope Serepa-Dlamini
Microbiol. Res. 2025, 16(9), 198; https://doi.org/10.3390/microbiolres16090198 - 1 Sep 2025
Abstract
Bacillus cereus AS_3 and Bacillus cereus AS_5 are bacterial endophytes isolated from sterilized leaves of the medical plant Alectra sessiliflora, which were previously identified using 16S rRNA sequencing. Here, we present the whole-genome sequencing and annotation of strains AS_3 and AS_5, the [...] Read more.
Bacillus cereus AS_3 and Bacillus cereus AS_5 are bacterial endophytes isolated from sterilized leaves of the medical plant Alectra sessiliflora, which were previously identified using 16S rRNA sequencing. Here, we present the whole-genome sequencing and annotation of strains AS_3 and AS_5, the first genome report of Bacillus cereus strains from A. sessiliflora. The genome of strain AS_3 has 59 contigs, 5 503 542 bp draft circular chromosome, an N50 of 211,274 bp, and an average G+C content of 35.2%; whereas strain AS_5 has 38 contigs, 5,510,121 bp draft circular chromosome, an N50 of 536,033 bp, and an average G+C content of 35.2%. A total of 5679 protein-coding genes, 62 genes coding for RNAs, and 122 pseudogenes in the strain AS_3 genome were identified by the National Center for Biotechnology Information Prokaryotic Annotation pipeline, whereas a total of 5688 gene protein-coding genes were identified in AS_5, with 60 genes coding for RNAs and 120 pseudogenes. Phenotypic analysis and whole-genome sequencing analysis showed that AS_3 and AS_5 share similar characteristics, including Gram-positive, motile, rod-shaped, and endospore-forming have shown a high sequence similarity with Bacillus cereus, type strain ATCC 14579T. Strains AS_3 and AS_5 had genomic digital DNA–DNA hybridization (dDDH) with the type strain Bacillus cereus ATCC 14579T of 85.8% and 86%, respectively, and average nucleotide identities (ANIs) of 98% and 98.01%, respectively. Phylogenomic analysis confirmed that strains AS_3 and AS_5 share very similar genomic and phenotypic characteristics, and are closely related to the type strain Bacillus cereus type strain ATCC 14579T, supporting their classification within the Bacillus cereus species. A total of 10 secondary metabolite gene clusters, including siderophore type petrobactin, terpene type molybdenum cofactor, non-ribosomal peptide synthetase (NRPS) type bacillibactin, and β-lactone type fengycin, were predicted using AntiSMASH software (version 5.0). Putative genes potentially involved in bioremediation and endophytic lifestyle were identified in the genome analysis. Genome sequencing of Bacillus cereus AS_3 and Bacillus cereus AS_5 has provided genomic information and demonstrated potential biotechnological applications. Full article
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24 pages, 2945 KB  
Article
Comprehensive Investigation of Qatar Soil Bacterial Diversity and Its Correlation with Soil Nutrients
by Muhammad Riaz Ejaz, Kareem Badr, Farzin Shabani, Zahoor Ul Hassan, Nabil Zouari, Roda Al-Thani and Samir Jaoua
Microbiol. Res. 2025, 16(9), 196; https://doi.org/10.3390/microbiolres16090196 - 1 Sep 2025
Abstract
Arid and semi-arid regions show distinctive bacterial groups important for the sustainability of ecosystems and soil health. This study aims to investigate how environmental factors across five Qatari soils influence the taxonomic composition of bacterial communities and their predicted functional roles using 16S [...] Read more.
Arid and semi-arid regions show distinctive bacterial groups important for the sustainability of ecosystems and soil health. This study aims to investigate how environmental factors across five Qatari soils influence the taxonomic composition of bacterial communities and their predicted functional roles using 16S rRNA amplicon sequencing and soil chemical analysis. Soil samples from five different locations in Qatar (three coastal and two inland) identified 26 bacterial phyla, which were dominated by Actinomycetota (35–43%), Pseudomonadota (12–16%), and Acidobacteriota (4–13%). Species-level analysis discovered taxa such as Rubrobacter tropicus, Longimicrobium terrae, Gaiella occulta, Kallotenue papyrolyticum, and Sphingomonas jaspsi, suggesting the presence of possible novel microbial families. The functional predictions showed development in pathways related to amino acid metabolism, carbohydrate metabolism, and stress tolerance. In addition, heavy-metal-related taxa, which are known to harbor genes for metal resistance mechanisms including efflux pumps, metal chelation, and oxidative stress tolerance. The presence of Streptomyces, Pseudomonas, and Bacillus highlights their roles in stress tolerance, biodegradation, and metabolite production. These findings improve the understanding of microbial roles in dry soils, especially in nutrient cycling and ecosystem resilience. They highlight the importance of local bacteria for sustaining desert soil functions. Further research is needed to validate these relationships, using metabolomic approaches while monitoring microbial-community-changing aspects under fluctuating environmental conditions. Full article
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17 pages, 3740 KB  
Article
Diversity Characteristics and Composition of Gut Microbiota in Antheraea pernyi (Lepidoptera: Saturniidae) Larvae Across Different Instars
by Peng Hou, Li Liu, Xin Ma, Ying Men, Ding Yang, Jianfeng Wang and Chuntian Zhang
Insects 2025, 16(9), 909; https://doi.org/10.3390/insects16090909 (registering DOI) - 1 Sep 2025
Abstract
Antheraea pernyi (Lepidoptera: Saturniidae) is an economically important silk-producing insect, whose gut microbiota play a crucial role in growth, development, and nutrient metabolism. This study focused on the entire larval developmental stages of A. pernyi. Using the Illumina MiSeq high-throughput sequencing platform, we [...] Read more.
Antheraea pernyi (Lepidoptera: Saturniidae) is an economically important silk-producing insect, whose gut microbiota play a crucial role in growth, development, and nutrient metabolism. This study focused on the entire larval developmental stages of A. pernyi. Using the Illumina MiSeq high-throughput sequencing platform, we performed 16S rRNA gene amplicon sequencing on the gut microbiota of laboratory-reared A. pernyi larvae, analyzing in detail the composition and diversity characteristics of the gut microbial communities across all five instars (1st to 5th instar). Additionally, functional predictions were conducted to explore the potential roles of these microbiota during larvae development. The study revealed that the core gut microbiota of A. pernyi larvae primarily consisted of Actinomycetota (39.78%), Cyanobacteriota (32.46%), Bacillota (18.08%), and Pseudomonadota (9.02%). Among these, Actinomycetota dominated in the 1st to 4th-instar larvae, while Cyanobacteriota became the predominant phylum in the 5th instar. Linear discriminant analysis effect size identified statistically significant biomarkers across different instar larvae of A. pernyi. Alpha diversity analysis showed that gut microbiota diversity initially increased and then decreased with larval development, peaking in the 3rd instar, and reaching its lowest level in the 5th instar. Principal coordinate analysis (PCoA) of beta diversity indicated that the gut microbiota structures of the 1st to 4th instars were similar but significantly differed from that of the 5th instar. Functional prediction analysis based on the KEGG database revealed that Carbohydrate metabolism and Amino acid metabolism-related genes were significantly lower in the 5th instar compared to other instars, while Energy metabolism and Cofactor and vitamin metabolism-related genes were significantly higher. This study offers valuable insights for the development of gut microbial resources in Lepidoptera insects. Full article
(This article belongs to the Section Insect Physiology, Reproduction and Development)
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13 pages, 288 KB  
Article
Genetic Characterization of Salmonella and Analysis of Ciprofloxacin Resistance Using Sanger Technique in Romania, 2024
by Elena Roxana Buzilă, Raluca Gatej, Cristina Trifan, Teodora Vremera, Mihaela Leustean, Adina David, Daniela Cosmina Bosogea, Georgiana Barbu, Adina Gatea, Ciprian Ilie and Luminița Smaranda Iancu
Bacteria 2025, 4(3), 43; https://doi.org/10.3390/bacteria4030043 (registering DOI) - 1 Sep 2025
Abstract
Salmonella is a major foodborne pathogen, representing a significant public health concern across the European Union (EU), accounting for 39% of foodborne illness-related hospitalizations in 2022, with the highest rates observed in Romania, Cyprus, Greece, and Lithuania. This pilot study aimed to enhance [...] Read more.
Salmonella is a major foodborne pathogen, representing a significant public health concern across the European Union (EU), accounting for 39% of foodborne illness-related hospitalizations in 2022, with the highest rates observed in Romania, Cyprus, Greece, and Lithuania. This pilot study aimed to enhance the surveillance and characterization of Salmonella by implementing both phenotypic and genotypic methods for strain typing, as well as for the detection and confirmation of resistance to ciprofloxacin. Materials and methods: A total of 109 Salmonella strains from acute diarrheal cases in North-Eastern Romania were collected (January–August 2024). From these, 19 representative isolates were selected for molecular characterization, including Multi-Locus Sequence Typing (MLST) and the detection of ciprofloxacin resistance determinants. Whole-Genome Sequencing (WGS) was subsequently performed to confirm serotype identity and resistance markers. Results: The 19 isolates underwent Multi-Locus Sequence Typing (MLST) and ciprofloxacin resistance profiling, with Whole-Genome Sequencing (WGS) for confirmation. MLST identified S. Enteritidis (42.1%) as the predominant serotype, followed by S. Typhimurium, S. Livingstone, and S. Infantis. WGS confirmed serotypes in 15 isolates; 2 showed discrepancies with phenotypic results. Phenotypic resistance to ciprofloxacin was detected in 12/19 (63.2%) of the isolates, 6/12 presenting gyrA mutations (S83Y, D87G), and 2/12 strains presenting the plasmid-mediated qnrB19 gene. Full article
17 pages, 3797 KB  
Article
Transcriptomic and Metabolomic Analyses Provide Insights into Cryptocaryon irritans Resistance in Golden Pompano (Trachinotus ovatus)
by Bo Liu, Bao-Suo Liu, Jing-Wen Yang, Hua-Yang Guo, Nan Zhang, Teng-Fei Zhu, Lin Xian, Ke-Cheng Zhu and Dian-Chang Zhang
Fishes 2025, 10(9), 426; https://doi.org/10.3390/fishes10090426 (registering DOI) - 1 Sep 2025
Abstract
Golden pompano (Trachinotus ovatus) is an economically important fish species along China’s southern coast. However, infections by Cryptocaryon irritans severely constrain the healthy and sustainable development of the aquaculture industry. To investigate the genetic basis of resistance to this parasite in [...] Read more.
Golden pompano (Trachinotus ovatus) is an economically important fish species along China’s southern coast. However, infections by Cryptocaryon irritans severely constrain the healthy and sustainable development of the aquaculture industry. To investigate the genetic basis of resistance to this parasite in golden pompano, this study employed transcriptomic and metabolomic analyses to compare differences between susceptible (ES) and resistant (RS) groups following C. irritans challenge. Transcriptome analysis identified 2031 differentially expressed genes (DEGs) between EST and RST groups, comprising 1004 up-regulated and 1027 down-regulated genes. Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment revealed that these DEGs were primarily enriched in lipid metabolism and amino acid metabolism pathways. Untargeted metabolomics detected 461 significantly differentially abundant metabolites (295 up-regulated, 166 down-regulated), confirming pronounced metabolic differences between ES and RS groups, particularly in lipid and amino acid metabolism. Further, KEGG enrichment highlighted steroid hormone biosynthesis, α-linolenic acid metabolism, and arachidonic acid metabolism as the most significantly altered pathways upon infection. This integrated transcriptomic and metabolomic study reveals substantial differences in gene expression and metabolite profiles between susceptible and resistant golden pompano in response to C. irritans. These changes predominantly involve lipid metabolism and amino acid metabolism, suggesting that these processes are critical in determining host resistance/susceptibility. Full article
(This article belongs to the Special Issue Molecular Mechanism of Fish Immune Response to Pathogens)
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13 pages, 1866 KB  
Article
Transcriptomic Analysis of Chilo suppressalis (Walker) (Lepidoptera: Pyralidae) Reveals Cold Tolerance Mechanisms Under Parasitism Stress
by Chuan-Lei Dong, Elyar Abil, Rong Ji, Yu-Zhou Du and Ming-Xing Lu
Insects 2025, 16(9), 907; https://doi.org/10.3390/insects16090907 (registering DOI) - 1 Sep 2025
Abstract
Parasitoids exhibit remarkable abilities to manipulate host physiology, ensuring offspring survival and development. This study investigated the molecular mechanisms underlying how the parasitoid Cotesia chilonis modulates cold tolerance in its host, the rice stem borer Chilo suppressalis, using transcriptome sequencing. We found [...] Read more.
Parasitoids exhibit remarkable abilities to manipulate host physiology, ensuring offspring survival and development. This study investigated the molecular mechanisms underlying how the parasitoid Cotesia chilonis modulates cold tolerance in its host, the rice stem borer Chilo suppressalis, using transcriptome sequencing. We found that the host larvae’s supercooling point was lowest at 3 days post-parasitism but increased significantly by day 4. Transcriptome analysis identified 507 differentially expressed genes (DEGs), including 235 up-regulated by parasitism. Functional enrichment revealed that these DEGs were primarily associated with ribosome biogenesis, protein processing in the endoplasmic reticulum (ER), and oxidative phosphorylation under parasitism stress. Notably, 24 DEGs linked to temperature tolerance were predominantly heat shock proteins (HSPs) and calcium signaling-related genes. The reliability of transcriptome data was confirmed via RT-qPCR for eight randomly selected DEGs. Functional assays demonstrated that parasitism stress significantly inhibited ER activity. However, HSP expression did not significantly affect ER activity or cytosolic Ca2+ concentration in the hemolymph cells of C. suppressalis larvae. This research provides insights into the complex physiological and molecular mechanisms through which C. suppressalis responds to parasitism stress, particularly concerning cold tolerance modulation. Full article
(This article belongs to the Special Issue Insect Transcriptomics)
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Article
A Programmable Finite-Replicated Organism Framework for Balanced Safety and Functionality
by Mengyuan Wang, Pei Du, Fankang Meng, Wenhui Zhang, Yanhui Xiang, Qiong Wu and Chunbo Lou
Life 2025, 15(9), 1381; https://doi.org/10.3390/life15091381 - 1 Sep 2025
Abstract
Live-attenuated vaccines face a critical challenge in balancing immunogenicity with safety. To address this, we engineered programmable finite-replicated organisms (FROs) by depositing a limited number of indispensable components (such as noncanonical amino acids, ncAAs) within the cell, consuming the coenabling precise control of [...] Read more.
Live-attenuated vaccines face a critical challenge in balancing immunogenicity with safety. To address this, we engineered programmable finite-replicated organisms (FROs) by depositing a limited number of indispensable components (such as noncanonical amino acids, ncAAs) within the cell, consuming the coenabling precise control of bacterial replication capability while preserving antigenic breadth. Two strategies were adopted to achieve the following purposes: (1) encoding ncAA in essential genes; (2) encoding ncAA in antitoxin of toxin–antitoxin (TA) systems. As noncanonical amino acids, 3,5-dichlorotyrosine (Cl2Y) was encoded by the amber codon (TAG) and inserted into the essential genes (e.g., serS, murG, and dnaA) or antitoxin genes. After optimizing expression and the number of amber codons in the storage genes, the FRO cells can grow up to six generations, achieving amplification approaching 100 times after depletion of the ncAA in the growth medium. The escape frequencies are 10−5 to 10−7, which need to be optimized by combining multiple storage genes in the same genome in the future. This work holds the potential to amplify the amounts of antigens for vaccines, potentially accelerating the development of next-generation vaccines against antibiotic-resistant threats. Full article
(This article belongs to the Special Issue Synthetic Genetic Elements, Devices, and Systems: 2nd Edition)
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