The western flower thrips Frankliniella occidentalis and the two-spotted spider mites Tetranychus urticae are common pests in agricultural production. To evaluate the potential of Amblyseius tamatavensis for controlling these two pests, the predatory preference, functional response, and intraspecific interference of A. tamatavensis towards the first and second instar nymphs of western flower thrips and various developmental stages of two-spotted spider mites were measured under controlled conditions. The experimental results showed that A. tamatavensis had a strong preference for the egg of two-spotted spider mites and the first instar nymphs of western flower thrips, with predation capacity a/Th of 128.21 and 7.837 respectively. The functional response of A. tamatavensis to all tested developmental stages of both pests conformed to the Holling type II model. In a limited space, the searching efficiency of A. tamatavensis decreased with increasing prey density. When space size and prey density were fixed, the average daily predation and predation efficiency (E) of the predatory mites significantly decreased as predator density increased, indicating pronounced intraspecific competition and interference. This study demonstrated that A. tamatavensis possessed strong predatory capability against T. urticae and F. occidentalis, highlighting its potential as a promising biological control agent.

At low temperatures, alterations in root-associated microorganisms enhance the antioxidant enzyme activity in plants, thereby increasing their stress resistance. Low-temperature stress is an important environmental factor that restricts plant growth and development. While microbial inoculants play a significant role in alleviating plant low-temperature stress, the specific mechanisms involved remain unclear. This study evaluated the effects of exogenously applied microbial inoculants (composed of Clostridium sensu stricto, Altererythrobacte, Gemmatimona, Stenotrophomona, and Leclerici) on tobacco seedling growth, antioxidant enzyme activity, and root surface microbial communities under low-temperature stress, as well as the interrelationships among these factors. Under low-temperature stress, plants were treated with increasing volumes of functional microbial agents at three levels (T1, T2, T3), compared to a control (CK) without inoculation. The results indicated that the application of microbial inoculants significantly promoted the growth of tobacco seedlings. Root growth and dry matter accumulation of tobacco seedlings were positively correlated with the amount of microbial inoculum applied. Furthermore, microbial inoculants significantly enhanced the activities of antioxidant enzymes (POD, CAT, and SOD) in tobacco seedlings. The T3 treatment increased these activities by 191.1

Pepino (Solanum muricatum), belonging to the family Solanaceae, is a perennial semi-woody herbaceous plant (Song et al. 2022) with significant economic value. However, in recent years, pepino has been threatened by various viral diseases. Viruses reported to infect pepino include Tobacco mosaic virus (TMV), Cucumber mosaic virus (CMV), Tomato mosaic virus (ToMV), Potato virus M (PVM), and Potato virus Y (PVY). These infections cause yield reduction and poor fruit quality, resulting in substantial economic losses to farmers. Pepper vein yellows virus (PeVYV), a member of the genus Polerovirus, is primarily transmitted by aphids in a non-propagative manner. Its natural hosts are mainly solanaceous plants, with pepper (Capsicum annuum) being the primary host. Infections have been reported globally in countries such as Spain, China, India, and Thailand (Zhao et al. 2021). In 2016, the first natural infection in tobacco (Nicotiana tabacum) by PeVYV was reported in Guizhou, China, causing interveinal yellowing and leaf crumpling symptoms in affected plants. During virus surveys in Mali and Benin, Africa, PeVYV was found infecting eggplant (Solanum melongena) (Wang et al.2016). Records from Spain and other regions also indicate that PeVYV infects black nightshade (Solanum nigrum) (Liu et al. 2019). As pepino (Solanum muricatum) is also a solanaceous plant, it is theoretically susceptible to solanaceous-infecting viruses. However, based on current literature and research, there are no public reports confirming that pepper vein yellows virus (PeVYV) can infect pepino (Solanum muricatum). In May 2023, pepino plants exhibiting leaf yellowing symptoms were observed at a plantation in Shilin County, Kunming City, Yunnan Province, China. Approximately 50% of the 100 plants inspected at the site showed symptoms of vein yellowing and mosaic mottling. To identify the causative virus, five symptomatic leaf samples from infected pepino plants collected in Shilin, Kunming were tested using a PeVYV-specific ELISA kit, with healthy pepino leaves serving as negative controls. The ELISA results showed that all samples tested were positive for PeVYV. To further confirm the presence of PeVYV in pepino, total RNA was extracted from at least one selected sample and subjected to RT-PCR using specific primers A 966-bp fragment, consistent with the expected size for PeVYV-YNs, was amplified from pepino samples. Sequencing yielded an 822-bp sequence. BLAST analysis revealed that PeVYV-YNs shared 92.13% and 91.92% nucleotide sequence identity with isolates Ca (MK931185) and YN (MT188667), respectively. The full genome sequence was obtained by RACE amplification. The isolate's genome is 5,988 nt in length (GenBank accession: OR906336) and exhibits a typical PeVYV genome structure. Phylogenetic analysis indicated that it forms a distinct branch, with nucleotide sequence similarity ranging from 79.7% to 98.4% compared to known isolates. In summary, while PeVYV infection and detection in pepper have been extensively studied, no prior reports of its infection in pepino exist. This study, by combining ELISA and RT-PCR with specific primers, demonstrates the natural infection of pepino by the PeVYV-YNs isolate. To our knowledge, this is the first report of pepper vein yellows virus infecting pepino. These findings provide a crucial basis for the prevention and control of viral diseases in pepino.

Background: The three commercial Enterovirus 71 (EV71) inactivated vaccines which have effectively controlled the EV71 pandemic currently rely on inherent variable in vivo potency methods for batch release. To align with 3R (Replacement, Reduction, Refinement) principles and enhance quality control, this study referred to WHO guidelines and the European Pharmacopoeia to develop in vitro relative potency (IVRP) methods. Methods: Working standards tracing to phase 3 clinical vaccines were established. Manufacture-specific IVRP methods were developed and validated per ICH Q14/Q2(R2), utilizing conformational epitope-targeting neutralizing monoclonal antibodies (MAbs). One of the MAbs (CT11F9) recognition sites was clarified with Cryo-EM. Subsequently, the performance of IVRP was assessed using varied concentrations and heat-treated vaccines. The correlation between IVRP and in vivo methods was analyzed, followed by setting IVRP specifications. Results: The manufacturer-specific working standard exhibited ED50 values comparable to those of related phase 3 clinical vaccines. All IVRP methods achieved a relative bias/precision/total error ≤ 15%. The IVRP methods correlated with in vivo methods (p < 0.05, r > 0.9) can discriminate EV71 antigen concentrations (p < 0.01, r > 0.99) and indicate the stability of the vaccines. Cryo-EM was adopted to identify the epitopes recognized by CT11F9, revealing that this neutralizing antibody recognizes a conformational epitope spanning VP1-3 of the same protomer. Using 31–47 batches of commercial vaccines, IVRP specifications were proposed as 0.56–1.35, 0.58–1.40, and 0.54–1.50. Conclusions: Based on conformational epitope-targeting neutralizing MAbs, manufacturer-specific IVRP methods, which were sensitive to process variations and correlated with in vivo results, have been established. IVRP methods provide a reliable, animal-free alternative for EV71 vaccine batch release.

UA V and IoT have gradually become the significant technology in smart agriculture. In this paper, a three-layer IoT -UA V system is proposed in smart agriculture. This proposed system is comprised of three distinct layers: 1) the bottom layer composed of GUs; 2) the middle layer composed of MEC servers carried by M-UA Vs and BS; 3) the top layer composed of the AUs. The middle layer provides computing service to the bottom and top layer to help them execute heavy computation tasks. Meanwhile, we study the energy consumption minimization problem by jointly optimizing the 3D trajectory of the M-UAVs and resource allocation. Our approach employs a combination of the block coordinate descent (BCD) technique and the successive convex approximation (SCA) technique. Through numerical analysis, our proposed scheme demonstrates superior performance compared to other baseline.

Chelonus munakatae Matsumura (Hymenoptera: Braconidae) is an important parasitoid of several lepidopteran species. The mitochondrial genome of this species is a crucial source for molecular identification and genetic studies. Here, we used next-generation sequencing to obtain the C . munakatae mitochondrial genome, which was found to be 14,868 bp in length and contained 13 protein-coding genes, 22 tRNAs and two rRNAs. The nucleotide composition exhibits a significant bias towards A + T content, with a value of 86.8 %. A large number of PCG and tRNA gene rearrangement events were detected ( I-V-M-C-W-Q and A-N-S1-E-R-F ), which represent a novel rearrangement type in Hymenoptera. Furthermore, synteny analysis based on the whole mitochondrial genomes showed that a significant rearrangement of homologous blocks had occurred within this Braconidae species. This study not only delineates the mitochondrial characteristics of C. munakatae but also significantly contributes to the repository of mitochondrial genetic information for the Braconidae family. Moreover, it highlights the occurrence of gene rearrangement within Hymenoptera species, offering valuable insights into the evolutionary dynamics and genomic variability of these insects.

目的:本文以对生物制品生物效价检测进行规范的《中国药典》2020年版新增9401指导原则为标准进行EV71抗原检测方法的验证,进一步浅析生物制品生物活性/效价检测的方法学验证.方法:依据《中国药典》2020年版9401指导原则,结合EV71抗原检测方法特点及验证目的对其进行专属性、相对准确度、精密度、线性和范围5个方面进行方法学验证.结果:EV71抗原检测方法专属性强,6个效价水平几何变异系数为4.7％-13.0％,均小于15％,相对偏倚为-6.05％～9.03％,均在±15％范围内,定量范围为225％～31％,线性回归相关系数为0.999 5,5方面验证均符合要求,且适用于该检测方法的使用.结论:本原则可具体指导生物学检测方法验证,在对不同检测方法进行验证时,根据检测目的、方法的原理、方法的技术特点、被检物的成分等设计具体方案.

Fusarium diseases include wilts, blights, rots, and cankers of many horticultural, field, ornamental, and forest crops in both agricultural and natural ecosystems, and they significantly hinder food plant production. Here, we describe a novel mycovirus, tentatively designated as "Fusarium fusarivirus 1" (FuFV1), which was discovered in an isolate of the phytopathogenic fungus Fusarium sp. FuFV1 has a positive-sense single-stranded RNA (+ssRNA) genome of 6,391 nucleotides (nt) containing three open reading frames (ORFs). ORF1 encodes a large polypeptide of 1,501 amino acids (aa) with conserved RNA-dependent RNA polymerase (RdRp) and helicase (Hel) domains. ORF2, overlapping ORF1 by 122 nucleotides, encodes a polypeptide with a conserved Smc domain. The third and smaller ORF (ORF3) encodes a polypeptide with an unknown function. BLASTp analysis of the ORF1-encoded polypeptide revealed that FuFV1 shares the highest aa sequence similarity (68.5% identity, E-value 0.0) with Fusarium poae fusarivirus 1 (FpFV1, genus Alphafusarivirus). Phylogenetic analysis of the RdRp and helicase (Hel) sequences indicated that FuFV1 clustered closely with FpFV1 in a separate branch within the clade containing members of the genus Alphafusarivirus. Based on these results, we propose that FuFV1 should be considered a novel mycovirus belonging to the genus Alphafusarivirus of the family Fusariviridae.

目的 比较干燥保存及湿保存肠道病毒71型(enterovirus 71,EV71)抗原检测板的稳定性.方法 用羊抗EV71多克隆抗体包被96孔酶标板,制备20块EV71抗原检测板,10块板置空气干燥后真空包装,-25℃保存;另外10块板用洗涤液浸泡,封口后置4℃保存.分别于保存0、20、40、60、90、120、180d时,检测5份EV71样品.结果 干燥保存EV71抗原检测板检测5份样品7个时间点的均值分别为315.0、299.4、7 795.3、827.6、261.4 U/mL,湿保存的EV71抗原检测板检测结果均值分别为354.3、347.1、6 766.6、749.7、277.3 U/mL,各时间点两种方法检测结果差异无统计学意义(P＞0.05).两种方法保存180 d时,EV71抗原检测板均可保持较好的活性(P＞0.05).结论 两种方法保存EV71抗原检测板6个月内,均具有良好的稳定性.

Objective To evaluate the recovery of polio virus Sabin strain of three types after dialysis.Methods Polio virus Sabin strain of types Ⅰ,Ⅱ and Ⅲ,at high titers,were dialyzed.The recoveries of titers of each type before and after dialysis,as well as the clearance rates of formaldehyde after dialysis,were compared.However,the polio virus at titers of 101,100 and 10-1 CCID50/mL,and the samples of the same type and titer before and after dialysis were inoculated to Hep-2 cells and subcultured for three passages.The cytopathic effect (CPE) rates and virus titers before and after dialysis were compared.Results The volume and titer of samples of various types at high titers after dialysis showed no significant difference with those before dialysis (P ＞ 0.05),while the clearance rate of formaldehyde after dialysis was more than 99％.All the samples of various types at a titer of 101 CCID50/mL caused CPE after subculture for three passages.The CPE rates caused by samples of types Ⅰ,Ⅱ and Ⅲ,each at a titer of 100 CCID50/mL,were 66％,33％and 33％ respectively after subculture for one passage,all of which were 100％ after subculture for two and three passages.However,the samples of various types at a titer of 10-1 CCID50/mL caused no CPE after subculture for three passages.All the virus titers of samples after CPE were more than 6.0 lgCCID50/mL.Conclusion The recovery of three types of polio virus Sabin strain of three types were high after dialysis,with little loss,indicating that dialysis might be used for purification of the virus.

Objective To compare 4-parameter model with parallel line assay method in the application in biological test statistics for antigens of Sabin inactivated poliovirus vaccine (sIPV) and enterovirus 71 (EV71). Methods The 4-parameter model and parallel line assay were methodologically validated using the detection data of sIPV-D antigen and EV71 antigen. Then, both methods were used to calculate the detection result of samples with different antigen content. Results The 4-parameter model was fit for sIPV-D antigen detection data (coefficient of determination 0.998, residual value ≤0.059), and parallel line analysis was reliable (regression: reference F=727.49, P 0.05). For EV71 antigen, 4-parameter model was appropriate (coefficient of determination 1.000, residual value≤0.025), and parallel line analysis was reliable (regression: reference F=126.63, P 0.05). There was no significant difference between the calculation results of two methods for different antigen content of sIPV and EV71 (t=0.248-1.023, all P>0.05). Conclusion Both 4-parameter model and parallel line assay can be applied in the ELISA detection system of sIPV-D and EV71 antigens. Key words: Models, statistical; Viral vaccines; Antigen, viral; Enzyme-linked immunosorbent assay

目的 评价CA16 K168/8疫苗候选株免疫血清对不同CA16毒株的交叉中和能力及对致乳鼠麻痹CA16临床分离株的体内中和保护能力.方法 将CA16 K168/8疫苗候选株病毒纯化液辅以弗氏佐剂经背部及侧腹部皮下多点注射免疫新西兰大白兔,制备高效价免疫血清.采用微量细胞病变法检测中和抗体效价;用中和抗体效价终浓度为32 U的特异性免疫血清对18株不同CA16临床分离毒株及15株EV71毒株进行交叉中和指数检测;选取中和抗体效价为16、32、64、128、256 U的特异性免疫血清对可致乳鼠产生明显麻痹作用的CA16 FY-18株进行体内中和保护水平检测.结果 获得的兔抗CA16 K168/8血清对14株CA16毒株的中和效价为1∶1 024～1∶6144,GMT值为1∶3 153;可于体外完全中和18株不同CA16毒株,中和指数为100～5 623.4,平均值为794.1,而对15株EV71临床分离株中和指数仅为0.32～5.62,平均值为2.2;体内中和保护水平随抗体效价升高而增加,呈量-效关系,当中和抗体效价达128 U时,可完全保护乳鼠不发生临床麻痹反应.结论 CA16 K168/8疫苗候选株特异性免疫血清体内、外试验结果表明其具有良好的免疫保护性.

目的 探究肠道病毒71型(EV-A71) FY23-K株在不同温度下连续传代对其生物学特性的影响,旨在获得EV-A71的减毒候选株.方法 在33℃条件下进行FY23-K株克隆筛选,之后分别在33 ℃及35℃下连续传代,并进行病毒生长增殖特性、乳鼠致病性及在小鼠体内的免疫原性检测,对比分析其各项生物学特性的变化.结果 挑选出感染性滴度较高的6个克隆,在33℃条件下连续传代后发现,其感染性滴度逐渐降低,且相同接种条件下,达到75％病变程度的收获时间随代次的增加逐渐延长,全26代后,接种至第8天细胞病变程度只达到40％.而在35 ℃条件下连续传代的病毒能够稳定增殖,保持良好的生长特性,毒株感染性滴度＞ 6.5 lg CCID50/ml;35℃传代后毒株的免疫原性良好,6株克隆株有4株样品的灭活抗原免疫小鼠后抗体的阳转率可达100.0％,GMT可达1∶128,最高的中和抗体效价为1∶512;活病毒免疫小鼠抗体阳转率达88.9％以上,中和抗体效价均＞1∶16;传代样品攻击乳鼠后,乳鼠无发病、死亡情况,其脑、脊髓也未产生明显病理性损伤,仅在肌肉组织细胞间隙发现少量的炎性细胞浸润,在脑前额叶出现数个胶质小结.结论 FY-23K克隆株在35℃低温连续传代后有稳定的生长特性和良好的免疫原性,对乳鼠未表现出明显致病性,可作为EV-A71减毒活疫苗进一步研究的候选毒株.

Human echovirus 11 (E-11), a member of the species Enterovirus B, frequently causes aseptic meningitis and hand, foot and mouth disease (HFMD). We determined the complete genome sequence of strain 520K/YN/CHN/2010, isolated from a subject with HFMD and aseptic meningitis in Yunnan Province, China, in 2010. The strain shared 78.8% and 81.1% nucleotide sequence similarity with prototype strain Gregory in the complete VP1 gene and the complete genome, respectively. Only the VP2-VP3-VP1 genome region of 520K/YN/CHN/2010 was similar to that of the E-11 strain; the other genome regions were most similar to those of other members of the species Enterovirus B. Using phylogenetic analysis and sequence comparisons of the complete VP1 gene, E-11 strains could be divided into five genogroups, and the 520K/YN/CHN/2010 strain was found to belong to genogroup A. Recombination analysis showed evidence of recombination with other member of the species Enterovirus B, especially the E-9 strain MSH/KM812/2010. Persistent surveillance of HFMD pathogens might help predict potential emerging viruses and related disease outbreaks.

Hand, foot and mouth disease (HFMD) has been prevalent in China since 2008. Enterovirus 71 (EV71) is a common causative agent of HFMD, and various strains of EV71 are prevalent worldwide. The EV71C4 subgenotype is the most endemic strain in China. However, few studies investigating the biological characteristics and pathogeneses of different C4 strains have been reported. Therefore, the current study investigated 19 clinical EV71 strains in neonatal ICR mice and neonatal rhesus monkeys by comparing pathogenicity; the virulence of different viral passages, dosages, and routes of infection; and the effects produced by subject animal age. These 19 clinical EV71 strains, which were of the same subtype, displayed varying pathogenic effects. Three strains (HE31, 231 and 262) induced limb paralysis in neonatal ICR mice. In addition, the degree of virulence was largely dependent upon the dose, route of infection, and number of passages of the challenge virus, as well as the ages of the infected animals. The present study provides valuable basic data to enable further research into EV71 pathogenesis and to facilitate the development of new drugs and vaccines.

Human coxsackievirus A9 (CVA9) is a member of Enterovirus B species and may cause aseptic meningitis. The complete genome analyses of two strains CVA9 A242/YN/CHN/2009 and A108/YN/CHN/2009 isolated from aseptic meningitis cases in Yunnan Province, China, in 2009 were performed. These two strains shared 81.3 and 80.7, 81.0 and 81.1 % nucleotide similarity with prototype strain Griggs in the VP1-encoding sequence and the complete genome sequence, respectively. Through phylogenetic analysis and homogeneity analysis for twenty-eight VP1-encoding sequences, CVA9 strains could be divided into four genotypes and the Chinese strains might belong to genotype D. Similarity plot and bootscanning analyses showed evidence of recombination with other EVB viruses. In conclusion, persistent surveillance of circulating enterovirus might help understand the enterovirus evolution.

Objective To analyze the biological characteristics of clinical isolates of coxsackievirus group A type 16(CA16),a major pathogen of hand,foot and mouth disease(HFMD)and lay a foundation of vaccine development. Methods CA16 strains were isolated and identified from 178 stool and throat swab specimens of suspected patients with HFMD in Kunming Region,Yunnan Province,China,of which the growth characteristics,plaque morphology and virulence to suckling mice were analyzed. Results Seven CA16 strains,KM1,KM15,KM154,KM165,KM168,KM263 and KM881,were isolated,all of which belonged to B1 subtype. The strains were proliferated rapidly in Vero cells, most of which reached peak values of proliferation 4 ~ 6 d after inoculation. However,the infectious titers of these strains were significantly different,which ranged from 4. 75 to 7. 78 lgCCID50/ ml. The morphologies of plaques formed by the strains cultured in Vero cells for the same days were different,of which those formed by KM15,KM154 and KM168 were round,in needle-like size,and with clear edges,while those formed by KM1,KM881,KM263 and KM165 were large, irregular and with blurry edges. All the strains showed strong virulence,with which the suckling mice injected i. c. were attacked 3 ~ 6 d later. Both the morbidity and mortality of mice challenged with the strains except KM168,KM154, KM15 were 100%. The strains caused lesions at various degrees in brain,heart,lungs,muscles,spinal cord,liver and other tissues of suckling mice,especially in brain and muscle tissues. Conclusion The clinical isolates of CA16 in this study showed good adaptability to Vero cells,clear plaques and strong virulence,most of which showed high infectious titers. The isolates may be used for study on pathogenic mechanism of CA16 and vaccine development.

目的 评价人呼吸道合胞病毒(HRSV) Long株的免疫原性,为进一步的研究疫苗提供实验数据.方法 将HRSV制备成纯化抗原,取40只ICR鼠,随机分为4组,分别用纯化HRSV、纯化HRSV+ Al(OH)3、并设0.01 mol/L PBS、Al(OH)3对照组,0天、28天免疫小鼠,采用中和试验检测血清中和抗体,流式检测7种细胞因子.结果 初免后28天,加Al(OH)3组与无Al(OH)3组抗体阳转率分别为60％和50％,抗体几何平均效价(GMT)分别为1∶4.49和1∶5.27;加强免疫后,Al(OH)3组与无Al(OH)3组小鼠血清抗体阳转率均达100％,抗体几何平均效价(GMT)分别为1∶17.15和1∶10.56,平均增长3.8倍和2.0倍;中和抗体在组间无差异,而组内初免与再免的中和抗体有统计学差异(P＜0.01).加Al(OH)3样品组与无Al(OH)3样品组,所检测7种细胞因子均诱导IL-6及TNF,与两个对照组间有统计学差异(P＜0.05),而另外5种细胞因子则无明显升高.结论 人呼吸道合胞病毒Long株可产生良好的免疫效应.