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We sequenced the complete mitochondrial genomes of Lycosa shansia, and Lycosa singoriensis by combining Sanger and next-generation sequencing methods and analyzed the sequenced genomes in order to explore the phylogenetic placement and the mitogenome composition and evolution of these species. The mitochondrial genome of L. shansia was 14,638 bp, whereas that of L. singoriensis was 13,686 bp. The type of genes and direction of the coding strand present in the mitogenomes were the same as those in other species of Lycosoidea, including two ribosomal RNA genes (rRNAs), 22 transfer RNA genes (tRNAs), and 13 protein-coding genes (PCGs). The mitogenomes of the two species exhibited negative AT and positive GC skews. This indicated that the nucleotide compositions of the mitogenomes of L. singoriensis and L. shansia tended to be T and G. Both the mean and median values of Ka/Ks of ATP8 were the highest among the 13 protein-coding genes, indicating that it might have evolved more rapidly than the other protein-coding genes in both species. ATP8 may have undergone more relaxed selection constraints and accumulated more mutations. In addition, many tRNAs lacked T and D stem loops; a few had no acceptor stems. The assessed species were recovered nested within Lycosidae with high support. The present findings will be useful for future studies on the mitogenome evolution of spiders.

In the so-called “struggle for existence” competition, the venomous animals developed a smart and effective strategy, envenomation, for predation and defense. Biochemical analysis revealed that animal venoms are chemical pools of proteinase, peptide toxins, and small organic molecules with various biological activities. Of them, peptide toxins are of great molecular diversity and possess the capacity to modulate the activity of ion channels, the second largest group of drug targets expressed on the cell membrane, which makes them a rich resource for developing peptide drug pioneers. The spider Lycosa coelestis (L. coelestis) commonly found in farmland in China is a dominant natural enemy of agricultural pests; however, its venom composition and activity were never explored. Herein, we conducted cDNA library and transcriptomic sequencing of the venom gland of L. coelestis, which identified 1131 high-quality expressed sequence tags (ESTs), grouped into three categories denoted as toxin-like ESTs (597, 52.79%), cellular component ESTs (357, 31.56%), and non-matched ESTs (177, 15.65%). These toxin-like ESTs encode 98 non-reductant toxins, which are artificially divided into 11 families based on their sequence homology and cysteine frameworks (2–14 cysteines forming 1–7 disulfide bonds to stabilize the toxin structure). Furthermore, RP-HPLC purification combined with off-line MALDI-TOF analysis have detected 147 different peptides physically existing in the venom of L. coelestis. Electrophysiology analysis confirmed that the venom preferably inhibits the voltage-gated calcium channels in rat dorsal root ganglion neurons. Altogether, the present study has added a great lot of new members to the spider toxin superfamily and built the foundation for characterizing novel active peptides in the L. coelestis venom.

Background/Objectives: This study investigates the structural and biophysical properties of the wild-type antimicrobial peptide LyeTx I, isolated from the venom of the spider Lycosa erythrognatha, and its analog LyeTx I-b, designed to enhance antibacterial activity, selectivity, and membrane interactions by the acetylation and increased amphipathicty. Methods: To understand the mechanisms behind these enhanced properties, comparative analyses of the structural, topological, biophysical, and thermodynamic aspects of the interactions between each peptide and phospholipid bilayers were evaluated. Both peptides were isotopically labeled with 2H3-Ala and 15N-Leu to facilitate structural studies via NMR spectroscopy. Results: Circular dichroism and solid-state NMR analyses revealed that, while both peptides adopt α-helical conformations in membrane mimetic environments, LyeTx I-b exhibits a more amphipathic and extended helical structure, which correlates with its enhanced membrane interaction. The thermodynamic properties of the peptide–membrane interactions were quantitatively evaluated in the presence of phospholipid bilayers using ITC and DSC, highlighting a greater propensity of LyeTx I-b to disrupt lipid vesicles. Calcein release studies reveal that both peptides cause vesicle disruption, although DLS measurements and TEM imaging indicate distinct effects on phospholipid vesicle organization. While LyeTx I-b permeabilizes anionic membrane retaining the vesicle integrity, LyeTx I promotes significant vesicle agglutination. Furthermore, DSC and calcein release assays indicate that LyeTx I-b exhibits significantly lower cytotoxicity toward eukaryotic membranes compared to LyeTx I, suggesting greater selectivity for bacterial membranes. Conclusions: Our findings provide insights into the structural and functional modifications that enhance the antimicrobial and therapeutic potential of LyeTx I-b, offering valuable guidance for the design of novel peptides targeting resistant bacterial infections and cancer.

Major concerns exist regarding the environmental and human health risks caused by exposure to heavy metals. Spiders are often used as a model in ecotoxicological studies to assess soil pollution. Here, we measured the bioaccumulation of copper (Cu) and lead (Pb) in spiders, Lycosa terrestris and Pardosa birmanica , by inductively coupled plasma mass spectrometry (ICP-MS). We investigated whether Cu and Pb accumulation differs according to different spider species, single versus combined metal exposure, and routes of exposure. Spiders were exposed to 10 mM CuSO 4 and 10 mM PbCl 2 solutions separately or in combination (10 mM + 10 mM) through different exposure routes (spiked soil and food) for 6 weeks. The effect of metals on the survival and body mass of exposed and unexposed (control) spiders was determined. We found that in both spider species, accumulation of metals increased with exposure time. In single metal exposure, Cu accumulation from food was higher than soil exposure in both spider species, whereas the opposite was observed for Pb. The simultaneous uptake of Cu and Pb significantly decreased from food and soil, respectively. Soil exposure caused more accumulation of metals in L. terrestris than P. birmanica . Metal exposure via contaminated food caused higher mortality compared to soil exposure. Body mass of both spider species was significantly decreased and negatively correlated with metal’s concentration. Overall, our results show that bioaccumulation efficiency of Cu and Pb differs significantly in spiders exposed to metal’s mixture compared to single metal exposure and is dependent on the exposure route, the type of metal, and spider species. More understanding of the effects of exposure to metal mixture and exposure routes is essential for designing and supporting risk assessment and ecological monitoring programs.

In recent years, there has been a growing interest in antimicrobial peptides as innovative antimicrobial agents for combating drug-resistant bacterial infections, particularly in the fields of biofilm control and eradication. In the present study, a novel cationic antimicrobial peptide, named LC-AMP-F1, was derived from the cDNA library of the Lycosa coelestis venom gland. The sequence, physicochemical properties and secondary structure of LC-AMP-F1 were predicted and studied. LC-AMP-F1 was tested for stability, cytotoxicity, drug resistance, antibacterial activity, and antibiofilm activity in vitro compared with melittin, a well-studied antimicrobial peptide. The findings indicated that LC-AMP-F1 exhibited inhibitory effects on the growth of various bacteria, including five strains of multidrug-resistant bacteria commonly found in clinical settings. Additionally, LC-AMP-F1 demonstrated effective inhibition of biofilm formation and disruption of mature biofilms. Furthermore, LC-AMP-F1 exhibited favorable stability, minimal hemolytic activity, and low toxicity towards different types of eukaryotic cells. Also, it was found that the combination of LC-AMP-F1 with conventional antibiotics exhibited either synergistic or additive therapeutic benefits. Concerning the antibacterial mechanism, scanning electron microscopy and SYTOX Green staining results showed that LC-AMP-F1 increased cell membrane permeability and swiftly disrupted bacterial cell membranes to exert its antibacterial effects. In summary, the findings and studies facilitated the development and clinical application of novel antimicrobial agents.

The spider Lycosa sinensis represents a burrowing wolf spider (family Lycosidae) widely distributed in the cotton region of northern China, whose venom is rich in various bioactive peptides. In previous study, we used a combination strategy of peptidomic and transcriptomic analyses to systematically screen and identify potential antimicrobial peptides (AMPs) in Lycosa sinensis venom that matched the α-helix structures. In this work, the three peptides (LS-AMP-E1, LS-AMP-F1, and LS-AMP-G1) were subjected to sequence analysis of the physicochemical properties and helical wheel projection, and then six common clinical pathogenic bacteria (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) with multiple drug-resistance were isolated and cultured for the evaluation and analysis of antimicrobial activity of these peptides. The results showed that two peptides (LS-AMP-E1 and LS-AMP-F1) had different inhibitory activity against six clinical drug-resistant bacteria; they can effectively inhibit the formation of biofilm and have no obvious hemolytic effect. Moreover, both LS-AMP-E1 and LS-AMP-F1 exhibited varying degrees of synergistic therapeutic effects with traditional antibiotics (azithromycin, erythromycin, and doxycycline), significantly reducing the working concentration of antibiotics and AMPs. In terms of antimicrobial mechanisms, LS-AMP-E1 and LS-AMP-F1 destroyed the integrity of bacterial cell membranes in a short period of time and completely inhibited bacterial growth within 10 min of action. Meanwhile, high concentrations of Mg2+ effectively reduced the antibacterial activity of LS-AMP-E1 and LS-AMP-F1. Together, it suggested that the two peptides interact directly on bacterial cell membranes. Taken together, bioinformatic and functional analyses in the present work sheds light on the structure–function relationships of LS-AMPs, and facilitates the discovery and clinical application of novel AMPs.

Rice was an important food crop that one of the cultivation was the jajar legowo planting system. However, the effect of the planting system on brown planthopper and predatory Lycosa has not been well studied, related to microclimate changes. This study aims to identify microclimate factors in the jajar legowo and their effects on brown leafhopper populations and Lycosa . This research was designed through field experiments in rice fields with the treatment of legowo planting system (2: 1) and conventional system as a check. The research variables consisted of temperature, humidity, light intensity, brown leafhopper population, and Lycosa population. The results of this study showed the highest humidity was at the end of the planting period. The temperature and light intensity recorded in jajar legowo planting systems tends higher. The population of Lycosa in the Jajar legowo planting system was highest in the 2 week after transplanting (WAT), while in the conventional planting, the system was found in the 1 WAT The highest brown plant hopper population was recorded at 3 WAT, while in jajar legowo at 2 WAT. The results of this study showed no significant effect of microclimate factors on the Lycosa population and brown leafhopper populations. The benefit of this research is knowing the effect of microclimate on lycosa and WBC populations in Jajar Legowo and Conventional rice planting systems.

The venoms of spiders from the RTA (retro-lateral tibia apophysis) clade contain diverse short linear peptides (SLPs) that offer a rich source of therapeutic candidates. Many of these peptides have insecticidal, antimicrobial and/or cytolytic activities, but their biological functions are unclear. Here, we explore the bioactivity of all known members of the A-family of SLPs previously identified in the venom of the Chinese wolf spider (Lycosa shansia). Our broad approach included an in silico analysis of physicochemical properties and bioactivity profiling for cytotoxic, antiviral, insecticidal and antibacterial activities. We found that most members of the A-family can form α-helices and resemble the antibacterial peptides found in frog poison. The peptides we tested showed no cytotoxic, antiviral or insecticidal activities but were able to reduce the growth of bacteria, including clinically relevant strains of Staphylococcus epidermidis and Listeria monocytogenes. The absence of insecticidal activity may suggest that these peptides have no role in prey capture, but their antibacterial activity may help to defend the venom gland against infection.

Triflumezopyrim (TMP), a mesoionic insecticide, is commonly used for controlling planthoppers in rice. However, the relationship between the TMP residue and toxicity against brown planthoppers (BPHs) has not been studied in detail. We are reporting the dissipation of TMP from rice plant and soil under field conditions. The median lethal dose and median lethal concentration were 0.036 ng per insect and 0.525 mg L −1 , respectively. TMP at recommended dose (25 g a.i. ha −1 ) recorded 1.25 live BPH per hill as against 25.5 per hill in control at 14 days after treatment. TMP was considered to be harmless to the natural enemies, namely, Cyrtorhinus lividipennis and Lycosa pseudoannulata in the rice ecosystem. The residue of TMP from rice plant and soil was estimated using the QuEChERS method using three different doses (12.5, 25, and 50 g a.i. ha −1 ). The limit of quantitation (LOQ) of TMP in plant and soil was 5 µg kg −1 and 1 µg kg −1 , respectively. The maximum content of TMP in soil was less than 1% that of plant content on day 1. The dissipation pattern of TMP both from plant and soil was better explained by the first-order double-exponential decay model (FODED) as compared to the first-order kinetic model. Overall, the half-lives of TMP were ranged from 2.21 to 3.02 days in plant tissues and 3.78 to 4.79 days in soil as per the FODED model. Based on the persistence and toxicity of TMP, we could conclude that TMP will be effective against BPH up to 7–10 days after application. Triflumezopyrim with reasonable persistence and high efficacy could be recommended as an alternate pesticide in BPH management in rice.

Background Toxoplasmosis, caused by Toxoplasma gondii , poses serious health issues for humans and animals. Individuals with impaired immune systems are more susceptible to severe toxoplasmosis. Pregnant women infected by T. gondii can face the possibility of birth defects and miscarriages. While pyrimethamine and sulfadiazine are commonly used drugs in clinical practice, concerns over their side effects and resistance are on the rise. A spider peptide XYP1 isolated from Lycosa coelestis had potent anti- T. gondii effects, but it had a high synthesis cost and strong cytotoxicity. Methods This study intended to modify XYP1 for producing derived peptides via amino acid truncation and substitution. The anti- T. gondii effect was evaluated by trypan blue staining assay and killing experiment of RH strain tachyzoites. The CCK8 and hemolysis assays were used to compare their safeties. The morphological changes of T. gondii were observed by scanning electron microscope and transmission electron microscope. In addition, the mechanism of XYP1 against T. gondii through RNA-sequencing was further explored. Results In vivo and in vitro experiments revealed that XYP1-18 and XYP1-18-1 had excellent anti- T. gondii activity with lower cytotoxicity and hemolysis activity than XYP1. XYP1, XYP1-18, and XYP1-18-1 were able to disrupt the surface membrane integrity of T. gondii tachyzoites, forming pores and causing the disruption of organelles. Furthermore, RNA-sequencing analysis indicated that XYP1 could stimulate the host immune response to effectively eliminate T. gondii and lessen the host’s inflammatory reaction. Conclusions XYP1-18 had lower cytotoxicity and hemolysis activity than XYP1, as well as significantly extending the survival time of the mice. XYP1 played a role in host inflammation and immune responses, revealing its potential mechanism. Our research provided valuable insights into the development and application of peptide-based drugs, offering novel strategies and directions for treating toxoplasmosis. Graphical Abstract