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The Sulfur Conversion Functional Microbial Communities in Biogas Liquid Can Participate in Coal Degradation. 沼气液中的硫转化功能微生物群落可参与煤炭降解。
Pub Date : 2024-08-26 DOI: 10.33073/pjm-2024-027
Yang Li, Zhong Liang, Xinyue Yan, Tianqi Qin, Zhaojun Wu, Chunshan Zheng

The addition of biogas liquid is a practical way to improve the yield of biological coalbed methane. The microbial composition in biogas liquid is complex, and whether it could participate in the sulfur conversion of coal remains unknown. In this study, sulfur conversion-related microbial communities were enriched from biogas liquid, which was dominated by genera Anaerosolibacter, Bacillus, Hydrogenispora, and Oxobacter. The co-culture of these groups with coal significantly changed the coal microbial community composition but did not increase the content of CH4 and H2S. The changed microbial communities mainly belonged to phyla Firmicutes, Proteobacteria, and Actinobacteriota, and increased the relative abundance of genera Bacillus, Thermicanus, Hydrogenispora, Oxobacter, Lutispora, Anaerovorax, Desulfurispora, Ruminiclostridium, and Fonticella. From the microscopic structure of coal, an increase in the number of holes and roughness on the surface of the coal was found but the change of surface functional groups was weak. In addition, the addition of S-related microbial communities increased the contents of phoxim, methylthiobenzoylglycine and glibornuride M5 in aromatic compounds, as well as the content of lauryl hydrogen sulfate in alkyl compounds. Furthermore, the dibenzothiophene degradation-related microbial communities included Bacillus, Brevibacillus, Brevundimonas, Burkholderia-Caballeronia-Paraburkholderia, and Thermicanus, which can break C-S bonds or disrupt benzene rings to degrade dibenzothiophene. In conclusion, the S-related microbial communities in biogas liquid could rebuild the coal microbial community and be involved in the conversion process of organic sulfur in coal.

添加沼气液是提高生物煤层气产量的一种实用方法。沼液中的微生物组成复杂,能否参与煤的硫转化仍是未知数。本研究从沼气液中富集了与硫转化相关的微生物群落,其中以厌氧菌属、芽孢杆菌属、氢孢属和牛杆菌属为主。这些菌群与煤共培养后,煤微生物群落组成发生了显著变化,但并未增加 CH4 和 H2S 的含量。改变后的微生物群落主要属于固相菌门、变形菌门和放线菌门,并增加了芽孢杆菌属、Thermicanus属、Hydrogenispora属、Oxobacter属、Lutispora属、Anaerovorax属、Desulfurispora属、Ruminiclostridium属和Fonticella属的相对丰度。从煤的微观结构来看,煤表面的孔洞数量和粗糙度有所增加,但表面官能团的变化较弱。此外,S 相关微生物群落的加入增加了芳香族化合物中的辛硫磷、甲硫苯甲酰甘氨酸和胶脲 M5 的含量,以及烷基化合物中月桂基硫酸氢酯的含量。此外,与二苯并噻吩降解相关的微生物群落包括芽孢杆菌(Bacillus)、芽孢杆菌(Brevibacillus)、芽孢杆菌(Brevundimonas)、伯克霍尔德菌(Burkholderia-Caballeronia-Paraburkholderia)和嗜热菌(Thermicanus),它们可以断裂 C-S 键或破坏苯环,从而降解二苯并噻吩。总之,沼气液中与 S 有关的微生物群落可以重建煤微生物群落,并参与煤中有机硫的转化过程。
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引用次数: 0
Screening and Characterization of Probiotics Isolated from Traditional Fermented Products of Ethnic-Minorities in Northwest China and Evaluation Replacing Antibiotics Breeding Effect in Broiler. 从中国西北少数民族传统发酵产品中分离的益生菌的筛选和特性鉴定及替代抗生素在肉鸡养殖中的效果评估
Pub Date : 2024-08-25 DOI: 10.33073/pjm-2024-025
Ze Ye, Bin Ji, Yinan Peng, Jie Song, Tingwei Zhao, Zhiye Wang

In this study, Lactobacillus fermentum DM7-6 (DM7-6), Lactobacillus plantarum DM9-7 (DM9-7), and Bacillus subtilis YF9-4 (YF9-4) were isolated from traditional fermented products. The survival rate of DM7-6, DM9-7, and YF9-4 in simulated intestinal gastric fluid reached 61.29%, 44.82%, and 55.26%, respectively. These strains had inhibition ability against common pathogens, and the inhibition zone diameters were more than 7 mm. Antioxidant tests showed these strains had good scavenging capacity for superoxide anion, hydroxyl radical and DPPH, and the total reduction capacity reached 65%. Then DM7-6, DM9-7 and YF9-4 were fed to broilers to study the effects on antioxidant capacity, immune response, biochemical indices, tissue morphology, and gut microbiota. 180 healthy broilers were allocated randomly into six experimental groups. SOD, GSH-Px, and T-AOC in broilers serum were detected, and the results showed probiotics significantly improve antioxidant capacity compared to CK group, while antibiotics showed the opposite result. Besides, IgA, IgM, IgG, TNF-α, and IL-2 indicated it could significantly improve immunity by adding probiotics in broilers diets. However, antibiotics reduced immunoglobulin levels and enhanced inflammation index. Biochemical indicators and tissue morphology showed probiotics had a protective effect on metabolic organs. Gut microbiota analysis proved antibiotics could significantly decrease microbial community diversity and increase the proportion of opportunistic pathogens, while probiotics could improve the diversity of gut microbiota and promote the colonization of beneficial microorganisms. In summary, probiotics DM7-6, DM9-7, and YF9-4 can improve the broiler's health by improving antioxidant capacity and immune function, regulating gut microbiota, and can be used as alternative probiotics for antibiotics-free breeding of broilers.

本研究从传统发酵产品中分离出了发酵乳杆菌 DM7-6(DM7-6)、植物乳杆菌 DM9-7(DM9-7)和枯草芽孢杆菌 YF9-4(YF9-4)。在模拟肠胃液中,DM7-6、DM9-7 和 YF9-4 的存活率分别达到 61.29%、44.82% 和 55.26%。这些菌株对常见病原体具有抑制能力,抑制区直径均大于 7 毫米。抗氧化试验表明,这些菌株对超氧阴离子、羟自由基和 DPPH 具有良好的清除能力,总还原能力达到 65%。然后用 DM7-6、DM9-7 和 YF9-4 喂养肉鸡,研究它们对肉鸡抗氧化能力、免疫反应、生化指标、组织形态和肠道微生物群的影响。180 只健康肉鸡被随机分配到六个实验组。结果表明,与 CK 组相比,益生菌能显著提高肉鸡的抗氧化能力,而抗生素组则相反。此外,IgA、IgM、IgG、TNF-α 和 IL-2 都表明,在肉鸡日粮中添加益生菌能显著提高免疫力。然而,抗生素会降低免疫球蛋白的水平并提高炎症指数。生化指标和组织形态显示益生菌对代谢器官有保护作用。肠道微生物群分析表明,抗生素能显著降低微生物群落的多样性,增加机会性病原体的比例,而益生菌则能改善肠道微生物群的多样性,促进有益微生物的定植。总之,益生菌 DM7-6、DM9-7 和 YF9-4 可通过提高抗氧化能力和免疫功能、调节肠道微生物群来改善肉鸡的健康状况,可作为肉鸡无抗生素养殖的替代益生菌。
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引用次数: 0
The Presence of Lactobacillus spp. and its Effect on the Occurrence of Other Microorganisms in the Reproductive Tract of Polish Women. 波兰妇女生殖道中乳酸杆菌的存在及其对其他微生物存在的影响。
Pub Date : 2024-08-25 DOI: 10.33073/pjm-2024-024
Aneta Kiecka, Katarzyna Rak, Joanna Białecka, Anna Białecka, Marian Szczepanik

Disorders of the vaginal microbiota can lead to many complications and affect fertility. This study evaluates the role of Lactobacillus in the vagina and its impact on the incidence of colonization by pathogenic microorganisms, analyzing the results of 1,039 women of reproductive age (18-49 years) who underwent a microbiological examination of the reproductive tract in 2020. Samples were examined by microscopy, culture, and NAAT. As the number of Lactobacillus increases, the chance of developing symptoms decreases. In fact, it has been shown that the higher the number of Lactobacillus spp. the less frequently Gardnerella vaginalis and Streptococcus group B are observed. As the concentration of Lactobacillus spp. increases by one category, the risk of G. vaginalis after adjustment to age and pH decreases by 80% (p < 0.001). Similarly, the correlation between Lactobacillus spp. and vaginal pH was shown. After adjustment to age, the odds of prevalence pH > 4.5 for people with Lactobacillus category higher 1 is 76% lower.

阴道微生物群紊乱可导致多种并发症并影响生育能力。本研究对 2020 年接受生殖道微生物检查的 1039 名育龄妇女(18-49 岁)的结果进行了分析,评估了乳酸杆菌在阴道中的作用及其对病原微生物定植发生率的影响。样本通过显微镜、培养和 NAAT 进行了检查。随着乳酸杆菌数量的增加,出现症状的几率也会降低。事实上,研究表明,乳酸杆菌的数量越多,阴道加德纳菌和 B 型链球菌的出现频率就越低。当乳酸杆菌的浓度增加一个等级时,根据年龄和酸碱度进行调整后,阴道加德纳菌的风险就会降低 80%(p < 0.001)。同样,乳酸杆菌与阴道 pH 值之间也存在相关性。在对年龄进行调整后,乳酸杆菌类别高于 1 的人患 pH > 4.5 的几率降低了 76%。
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引用次数: 0
Cloning Cellulase Genes from Victoria Falls Rainforest Decaying Logs Metagenome. 克隆维多利亚瀑布雨林腐木元基因组中的纤维素酶基因。
Pub Date : 2024-07-29 DOI: 10.33073/pjm-2024-029
Makhosazana Nyathi, Zephaniah Dhlamini, Thembekile Ncube

The Victoria Falls rainforest is a protected site whose forest floors harbor a host of cellulolytic microorganisms involved in biomass degradation. This study collected decaying logs and soil from the rainforest for bioprospecting cellulases from their metagenomes. Metagenomic DNA was isolated from the compound sample. Degenerate cellulase primers were used to amplify cellulase genes in the metagenome. The resulting amplicons cloned into Z-competent Escherichia coli DH5α were analyzed by functional screening for the production of cellulase extracellularly. Functional screening of the clones resulted in one clone (Clone-i) testing positive for extracellular cellulase production. Submerged fermentation of Clone-i was carried out for cellulase production. The cellulases were characterized to determine their activity's optimum pH and temperature. The diversity of the cellulases produced by Clone-i was determined. Clone-i's optimum enzyme activity was observed after 72 hours of incubation at 50°C and pH 5. Clone-i produced 80% more exoglucanases as compared to endoglucanases. The cellulolytic Clone-i' isolate shows Victoria Falls rainforest's potential as an enzyme bioprospecting site, reflecting that metagenomics is a valuable tool in microbial ecology.

维多利亚瀑布热带雨林是一个受保护的地方,其森林地面蕴藏着大量参与生物质降解的纤维素分解微生物。这项研究收集了雨林中的腐烂原木和土壤,以便从它们的元基因组中对纤维素酶进行生物勘探。从复合样本中分离出元基因组 DNA。使用退化纤维素酶引物扩增元基因组中的纤维素酶基因。将得到的扩增子克隆到 Z 型大肠杆菌 DH5α 中,通过功能筛选分析细胞外纤维素酶的生产情况。对克隆进行功能筛选的结果是,一个克隆(克隆-i)在胞外纤维素酶生产方面呈阳性。对克隆-i 进行了浸没式发酵,以生产纤维素酶。对纤维素酶进行了表征,以确定其活性的最适 pH 值和温度。确定了 Clone-i 生产的纤维素酶的多样性。在 50°C 和 pH 值为 5 的条件下培养 72 小时后,Clone-i 的酶活性达到最佳。纤维素溶解克隆-i'分离物显示了维多利亚瀑布雨林作为酶生物勘探地点的潜力,反映了元基因组学是微生物生态学的一种宝贵工具。
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引用次数: 0
Guanylate-Binding Protein 1 (GBP1) Enhances IFN-α Mediated Antiviral Activity against Hepatitis B Virus Infection. 鸟苷酸结合蛋白 1 (GBP1) 可增强 IFN-α 介导的乙型肝炎病毒感染抗病毒活性。
Pub Date : 2024-06-20 eCollection Date: 2024-06-01 DOI: 10.33073/pjm-2024-021
Yadi Li, Haiying Luo, Xiaoxia Hu, Jiaojiao Gong, Guili Tan, Huating Luo, Rui Wang, Hao Pang, Renjie Yu, Bo Qin

Interferon-alpha (IFN-α) is a first-line drug for treating chronic hepatitis B (CHB). Guanylate-binding protein 1 (GBP1) is one of the interferon-stimulating factors, which participates in the innate immunity of the host and plays an antiviral and antibacterial role. In this study, we explored how GBP1 is involved in IFN-α antiviral activity against HBV. Before being gathered, HepG2-NTCP and HepG2 2.15 cells were transfected with the wild-type hGBP1 plasmid or si-GBP1, respectively, and followed by stimulation with Peg-IFNα-2b. We systematically explored the role of GBP1 in regulating HBV infection in cell models. Additionally, we also examined GBP1 levels in CHB patients. GBP1 activity increased, and its half-life was prolonged after HBV infection. Overexpression of GBP1 inhibited the production of HBsAg and HBeAg, as well as HBs protein and HBV total RNA levels, whereas silencing of GBP1 inhibited its ability to block viral infections. Interestingly, overexpressing GBP1 co-treatment with Peg-IFNα-2b further increased the antiviral effect of IFN-α, while GBP1 silencing co-treatment with Peg-IFNα-2b partly restored its inhibitory effect on HBV. Mechanistically, GBP1 mediates the anti-HBV response of Peg-IFNα-2b by targeting HBs. Analysis of clinical samples revealed that GBP1 was elevated in CHB patients and increased with Peg-IFNα-2b treatment, while GBP1 showed good stability in the interferon response group. Our study demonstrates that GBP1 inhibits HBV replication and promotes HBsAg clearance. It is possible to achieve antiviral effects through the regulation of IFN-α induced immune responses in response to HBV.

干扰素-α(IFN-α)是治疗慢性乙型肝炎(CHB)的一线药物。鸟苷酸结合蛋白 1(GBP1)是干扰素刺激因子之一,它参与宿主的先天免疫,发挥抗病毒和抗菌作用。本研究探讨了 GBP1 如何参与 IFN-α 对 HBV 的抗病毒活性。在收集前,分别用野生型 hGBP1 质粒或 si-GBP1 转染 HepG2-NTCP 和 HepG2 2.15 细胞,然后用 Peg-IFNα-2b 刺激。我们系统地探讨了 GBP1 在细胞模型中调控 HBV 感染的作用。此外,我们还检测了慢性阻塞性肺病患者体内的 GBP1 水平。在感染 HBV 后,GBP1 的活性增加,半衰期延长。过表达 GBP1 可抑制 HBsAg 和 HBeAg 的产生以及 HBs 蛋白和 HBV 总 RNA 水平,而沉默 GBP1 则会抑制其阻断病毒感染的能力。有趣的是,过表达 GBP1 并与 Peg-IFNα-2b 联合治疗可进一步增强 IFN-α 的抗病毒作用,而沉默 GBP1 并与 Peg-IFNα-2b 联合治疗可部分恢复其对 HBV 的抑制作用。从机理上讲,GBP1通过靶向HBs介导了Peg-IFNα-2b的抗HBV反应。临床样本分析显示,GBP1在CHB患者中升高,并随Peg-IFNα-2b治疗而升高,而GBP1在干扰素反应组中表现出良好的稳定性。我们的研究表明,GBP1 可抑制 HBV 复制并促进 HBsAg 清除。通过调节 IFN-α 诱导的对 HBV 的免疫反应可以达到抗病毒效果。
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引用次数: 0
Halogenated Secondary Metabolites from Higher Plants: Potent Drug Candidates for Chikungunya Using in silico Approaches. 高等植物中的卤代次生代谢物:利用硅学方法研究治疗基孔肯雅病的有效候选药物。
Pub Date : 2024-06-20 eCollection Date: 2024-06-01 DOI: 10.33073/pjm-2024-020
Sushil Kumar, Nidhi Joshi, Gourav Choudhir, Sakshi Sharma, Abhay Tiwari, Sulaiman Ali Alharbi, Saleh Alfarraj, Mohammad Javed Ansari

Chikungunya virus (CHIKV) causes a debilitating fever and joint pain, with no specific antiviral treatment available. Halogenated secondary metabolites from plants are a promising new class of drug candidates against chikungunya, with unique properties that make them effective against the virus. Plants produce these compounds to defend themselves against pests and pathogens, and they are effective against a wide range of viruses, including chikungunya. This study investigated the interactions of halogenated secondary metabolites with nsP2pro, a therapeutic target for CHIKV. A library of sixty-six halogenated plant metabolites screened previously for ADME properties was used. Metabolites without violation of Lipinski's rule were docked with nsP2pro using AutoDock Vina. To find the stability of the pipoxide chlorohydrin-nsP2pro complex, the GROMACS suite was used for MD simulation. The binding free energy of the ligand-protein complex was computed using MMPBSA. Molecular docking studies revealed that halogenated metabolites interact with nsP2pro, suggesting they are possible inhibitors. Pipoxide chlorohydrin showed the greatest affinity to the target. This was further confirmed by the MD simulations, surface accessible area, and MMPBSA studies. Pipoxide chlorohydrin, a halogenated metabolite, was the most potent against nsP2pro in the survey.

基孔肯雅病毒(CHIKV)会导致发烧和关节疼痛,使人衰弱,目前还没有特效的抗病毒治疗方法。植物中的卤代次生代谢物是一种很有前景的抗基孔肯雅病毒候选药物,其独特的性质使它们能够有效地抗病毒。植物产生这些化合物是为了抵御害虫和病原体,它们对包括基孔肯雅病毒在内的多种病毒都有效。本研究调查了卤代次生代谢物与 nsP2pro 的相互作用,nsP2pro 是 CHIKV 的治疗靶点。研究使用了一个包含 66 种卤代植物代谢物的文库,该文库之前已对其 ADME 特性进行了筛选。使用 AutoDock Vina 将不违反利宾斯基规则的代谢物与 nsP2pro 进行对接。为了确定哌啶氯醇-nsP2pro 复合物的稳定性,使用了 GROMACS 套件进行 MD 模拟。使用 MMPBSA 计算了配体-蛋白质复合物的结合自由能。分子对接研究发现,卤代代谢物与 nsP2pro 发生了相互作用,表明它们可能是抑制剂。硫代氯醇与目标物的亲和力最大。MD 模拟、表面可及面积和 MMPBSA 研究进一步证实了这一点。在此次调查中,卤代代谢物 Pipoxide chlorohydrin 对 nsP2pro 的抑制作用最强。
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引用次数: 0
Potential Usefulness of IgA for the Early Detection of SARS-CoV-2 Infection: Comparison With IgM. IgA 对早期检测 SARS-CoV-2 感染的潜在作用:与 IgM 的比较
Pub Date : 2024-06-20 eCollection Date: 2024-06-01 DOI: 10.33073/pjm-2024-019
Pei Wang

Serological testing can be a powerful complementary approach to achieve timely diagnosis of severe acute respiratory coronavirus 2 (SARS-CoV-2) infection, along with nucleic acid detection. Immunoglobulin (Ig) A antibodies are less frequently utilized to detect SARS-CoV-2 infection than IgM and IgG antibodies, even though IgA antibodies play an important role in protective immunity against SARS-CoV-2. This review discusses the differences in kinetics and assay performance between IgA and IgM antibodies and the factors influencing antibody responses. It highlights the potential usefulness of analyzing IgA antibodies for the early detection of SARS-CoV-2 infection. The early appearance of IgA and the high sensitivity of IgA-based immunoassays can aid in diagnosing coronavirus disease 2019. However, because of cross-reactivity, it is important to recognize the only moderate specificity of the early detection of SARS-CoV-2 IgA antibodies against spike antigens. Either the analysis of antibodies targeting the nucleocapsid antigen or a combination of antibodies against the nucleocapsid and spike antigens may strengthen the accuracy of serological evaluation.

血清学检测可以与核酸检测一起成为及时诊断严重急性呼吸道冠状病毒 2(SARS-CoV-2)感染的有力补充方法。与 IgM 和 IgG 抗体相比,免疫球蛋白 (Ig) A 抗体较少用于检测 SARS-CoV-2 感染,尽管 IgA 抗体在针对 SARS-CoV-2 的保护性免疫中发挥着重要作用。本综述讨论了 IgA 和 IgM 抗体在动力学和检测性能方面的差异,以及影响抗体反应的因素。它强调了分析 IgA 抗体对早期检测 SARS-CoV-2 感染的潜在作用。IgA 的早期出现和基于 IgA 的免疫测定的高灵敏度有助于 2019 年冠状病毒疾病的诊断。然而,由于存在交叉反应,必须认识到早期检测针对尖峰抗原的 SARS-CoV-2 IgA 抗体仅具有中等特异性。无论是分析针对核壳抗原的抗体,还是将针对核壳抗原和尖峰抗原的抗体结合起来,都可能提高血清学评估的准确性。
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引用次数: 0
Negative Pressure Level and Effects on Bacterial Growth Kinetics in an in vitro Wound Model. 负压水平及其对体外伤口模型中细菌生长动力学的影响
Pub Date : 2024-06-20 eCollection Date: 2024-06-01 DOI: 10.33073/pjm-2024-018
Adam Bobkiewicz, Wojciech Francuzik, Amy Martinkosky, Maciej Borejsza-Wysocki, Witold Ledwosinski, Krzysztof Szmyt, Tomasz Banasiewicz, Lukasz Krokowicz

Negative Pressure Wound Therapy (NPWT) has been widely adopted in wound healing strategies due to its multimodal mechanism of action. While NPWT's positive impression on wound healing is well-established, its effect on bacterial load reduction remains equivocal. This study investigates NPWT's efficacy in reducing bioburden using an in vitro porcine skin model, focusing on the impact of Staphylococcus aureus and Staphylococcus epidermidis. Custom-made negative pressure chambers were employed to apply varying negative pressures. Porcine skin was cut into 5 × 5 cm squares and three standardized wounds of 6 mm each were created using a biopsy punch. Then, wounds were infected with S. aureus and S. epidermidis bacterial suspensions diluted 1:10,000 to obtain a final concentration of 1.5 × 104 CFU/ml and were placed in negative pressure chambers. After incubation, bacterial counts were expressed as colony-forming units (CFU) per ml. For S. aureus at 120 hours, the median CFU, mean area per colony, and total growth area were notably lower at -80 mmHg when compared to -250 mmHg and -50 mmHg, suggesting an optimal negative pressure for the pressure-dependent inhibition of the bacterial proliferation. While analyzing S. epidermidis at 120 hours, the response to the negative pressure was similar but less clear, with the minor CFU at -100 mmHg. The influence of intermittent negative pressure on the S. epidermidis growth showed notably lower median CFU with the interval therapy every hour compared to the S. aureus control group. This study contributes valuable insights into NPWT's influence on the bacterial load, emphasizing the need for further research to reformulate its role in managing contaminated wounds.

负压伤口疗法(NPWT)因其多模式的作用机制而在伤口愈合策略中被广泛采用。虽然 NPWT 对伤口愈合的积极作用已得到证实,但其对减少细菌负荷的效果仍不明确。本研究使用体外猪皮肤模型研究了 NPWT 在减少生物负载方面的功效,重点关注金黄色葡萄球菌和表皮葡萄球菌的影响。采用定制的负压室来施加不同的负压。猪皮肤被切割成 5 × 5 厘米的正方形,并用活检打孔器造成三个各 6 毫米的标准化伤口。然后,将金黄色葡萄球菌和表皮葡萄球菌细菌悬浮液按 1:10,000 的比例稀释至最终浓度为 1.5 × 104 CFU/ml,并将其置于负压室中。培养后,细菌计数以每毫升菌落形成单位(CFU)表示。对于 120 小时的金黄色葡萄球菌,与 -250 mmHg 和 -50 mmHg 相比,-80 mmHg 下的 CFU 中位数、每个菌落的平均面积和总生长面积都明显较低,这表明压力依赖性抑制细菌增殖的最佳负压值。在分析 120 小时的表皮葡萄球菌时,对负压的反应类似,但不太明显,在-100 mmHg 时的 CFU 较少。间歇负压对表皮葡萄球菌生长的影响显示,与金黄色葡萄球菌对照组相比,每隔一小时治疗一次,表皮葡萄球菌的 CFU 中位数明显降低。这项研究为了解 NPWT 对细菌负荷的影响提供了宝贵的见解,强调了进一步研究其在管理污染伤口中的作用的必要性。
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引用次数: 0
Biological Characteristics of the Mycelium and Optimization of the Culture Medium for Phallus dongsun. 东荪菌丝的生物学特征和培养基的优化
Pub Date : 2024-06-20 eCollection Date: 2024-06-01 DOI: 10.33073/pjm-2024-022
Chao Kang, Xuan Zheng, Wankun Wang, Weijun Zeng, Jing Wang, Zhongxuan Liu, Ling Yang, Fang Wang, Yan Zhu

This study aimed to elucidate the influence of various culture medium components, including carbon sources, nitrogen sources, inorganic salts, suspension agents, and temperature, on the mycelial growth characteristics of Phallus dongsun. Employing single-factor experiments and response surface methodology within glass Petri dishes, the research identified that carrot powder, soybean powder, and ZnSO4 notably enhanced the proliferation of aerial mycelium, significantly augmenting the growth rate of P. dongsun mycelium. The resultant mycelium was observed to be dense, robust, and fluffy in texture. In particular, ZnSO4 markedly accelerated the mycelium growth rate. Furthermore, xanthan gum was found to effectively modulate the medium's viscosity, ensuring a stable suspension and facilitating nutrient equilibrium. The optimal cultivation temperature was determined to be 25°C, with mycelial growth ceasing below 5°C and mycelium perishing at temperatures exceeding 35°C. The optimal medium composition was established as follows: wheat starch 5 g/l, carrot powder 5 g/l, soybean powder 7.50 g/l, glucose 10 g/l, ZnSO4 0.71 g/l, NH4Cl 0.68 g/l, xanthan gum 0.5 g/l, and agar 20 g/l. Under these optimized conditions, the mycelium of P. dongsun exhibited a rapid growth rate (1.04 ± 0.14 mm/day), characterized by a thick, dense, and well-developed structure. This investigation provides a theoretical foundation for the conservation, strain selection, and breeding of P. dongsun.

本研究旨在阐明各种培养基成分(包括碳源、氮源、无机盐、悬浮剂和温度)对东胜菇菌丝生长特性的影响。通过在玻璃培养皿中进行单因素实验和响应面法,研究发现胡萝卜粉、大豆粉和 ZnSO4 能明显促进气生菌丝的增殖,显著提高东胜菇菌丝的生长速度。据观察,所产生的菌丝致密、粗壮、质地蓬松。特别是,硫酸锌明显加快了菌丝的生长速度。此外,研究还发现黄原胶能有效调节培养基的粘度,确保悬浮液稳定,促进营养平衡。最佳培养温度被确定为 25°C,低于 5°C 时菌丝停止生长,超过 35°C 时菌丝死亡。最佳培养基成分确定如下:小麦淀粉 5 克/升,胡萝卜粉 5 克/升,大豆粉 7.50 克/升,葡萄糖 10 克/升,ZnSO4 0.71 克/升,NH4Cl 0.68 克/升,黄原胶 0.5 克/升,琼脂 20 克/升。在这些优化条件下,东胜菇菌丝的生长速度很快(1.04 ± 0.14 毫米/天),其特点是粗壮、致密和结构发达。这项研究为东胜菇的保护、菌种选择和育种提供了理论依据。
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引用次数: 0
Establishment of a Nucleic Acid Detection Method for Norovirus GII.2 Genotype Based on RT-RPA and CRISPR/Cas12a-LFS. 基于 RT-RPA 和 CRISPR/Cas12a-LFS 的诺罗病毒 GII.2 基因型核酸检测方法的建立
Pub Date : 2024-06-20 eCollection Date: 2024-06-01 DOI: 10.33073/pjm-2024-023
Ting Wang, Hao Zeng, Jie Kang, Lanlan Lei, Jing Liu, Yuhong Zheng, Weidong Qian, Cheng Fan

To establish a rapid detection method for norovirus GII.2 genotype, this study employed reverse transcription recombinase polymerase amplification (RT-RPA) combined with CRISPR/Cas12a and lateral flow strip (RT-RPA-Cas12a-LFS). Here, the genome of norovirus GII.2 genotype was compared to identify highly conserved sequences, facilitating the design of RT-RPA primers and crRNA specific to the conserved regions of norovirus GII.2. Subsequently, the reaction parameters of RT-RPA were optimized and evaluated using agar-gel electrophoresis and LFS. The results indicate that the conserved sequences of norovirus GII.2 were successfully amplified through RT-RPA at 37°C for 25 minutes. Additionally, CRISPR/Cas12a-mediated cleavage detection was achieved through LFS at 37°C within 10 minutes using the amplification products as templates. Including the isothermal amplification reaction time, the total time is 35 minutes. The established RT-RPA-Cas12a-LFS method demonstrated specific detection of norovirus GII.2, yielding negative results for other viral genomes, and exhibited an excellent detection limit of 10 copies/μl. The RT-RPA-Cas12a-LFS method was further compared with qRT-PCR by analyzing 60 food-contaminated samples. The positive conformity rate was 100%, the negative conformity rate was 95.45%, and the overall conformity rate reached 98.33%. This detection method for norovirus GII.2 genotype is cost-effective, highly sensitive, specific, and easy to operate, offering a promising technical solution for field-based detection of the norovirus GII.2 genotype.

为了建立诺如病毒GII.2基因型的快速检测方法,本研究采用了反转录重组聚合酶扩增(RT-RPA)结合CRISPR/Cas12a和侧流条带(RT-RPA-Cas12a-LFS)的方法。通过比较诺如病毒GII.2基因型的基因组,确定了高度保守的序列,从而设计出针对诺如病毒GII.2保守区的特异性RT-RPA引物和crRNA。随后,对 RT-RPA 的反应参数进行了优化,并使用琼脂凝胶电泳和 LFS 进行了评估。结果表明,诺如病毒GII.2的保守序列在37℃、25分钟的RT-RPA反应中被成功扩增。此外,以扩增产物为模板,在 37°C 下 10 分钟内通过 LFS 实现了 CRISPR/Cas12a 介导的裂解检测。包括等温扩增反应时间在内,总时间为 35 分钟。已建立的 RT-RPA-Cas12a-LFS 方法能特异性检测诺如病毒 GII.2,对其他病毒基因组的检测结果为阴性,检测限为 10 拷贝/μl。通过分析 60 份食品污染样本,进一步比较了 RT-RPA-Cas12a-LFS 方法与 qRT-PCR 方法。阳性符合率为 100%,阴性符合率为 95.45%,总体符合率达到 98.33%。这种诺如病毒 GII.2 基因型的检测方法具有成本低、灵敏度高、特异性强、操作简便等特点,为现场检测诺如病毒 GII.2 基因型提供了一种可行的技术方案。
{"title":"Establishment of a Nucleic Acid Detection Method for Norovirus GII.2 Genotype Based on RT-RPA and CRISPR/Cas12a-LFS.","authors":"Ting Wang, Hao Zeng, Jie Kang, Lanlan Lei, Jing Liu, Yuhong Zheng, Weidong Qian, Cheng Fan","doi":"10.33073/pjm-2024-023","DOIUrl":"10.33073/pjm-2024-023","url":null,"abstract":"<p><p>To establish a rapid detection method for norovirus GII.2 genotype, this study employed reverse transcription recombinase polymerase amplification (RT-RPA) combined with CRISPR/Cas12a and lateral flow strip (RT-RPA-Cas12a-LFS). Here, the genome of norovirus GII.2 genotype was compared to identify highly conserved sequences, facilitating the design of RT-RPA primers and crRNA specific to the conserved regions of norovirus GII.2. Subsequently, the reaction parameters of RT-RPA were optimized and evaluated using agar-gel electrophoresis and LFS. The results indicate that the conserved sequences of norovirus GII.2 were successfully amplified through RT-RPA at 37°C for 25 minutes. Additionally, CRISPR/Cas12a-mediated cleavage detection was achieved through LFS at 37°C within 10 minutes using the amplification products as templates. Including the isothermal amplification reaction time, the total time is 35 minutes. The established RT-RPA-Cas12a-LFS method demonstrated specific detection of norovirus GII.2, yielding negative results for other viral genomes, and exhibited an excellent detection limit of 10 copies/μl. The RT-RPA-Cas12a-LFS method was further compared with qRT-PCR by analyzing 60 food-contaminated samples. The positive conformity rate was 100%, the negative conformity rate was 95.45%, and the overall conformity rate reached 98.33%. This detection method for norovirus GII.2 genotype is cost-effective, highly sensitive, specific, and easy to operate, offering a promising technical solution for field-based detection of the norovirus GII.2 genotype.</p>","PeriodicalId":94173,"journal":{"name":"Polish journal of microbiology","volume":"73 2","pages":"253-262"},"PeriodicalIF":0.0,"publicationDate":"2024-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11192556/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141437979","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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Polish journal of microbiology
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