A molecular tool to identify Anastatus parasitoids of the brown marmorated stink bug

Globally, Anastatus species (Hymenoptera: Eupelmidae) are associated with the invasive agricultural pest Halyomorpha halys (Stål) (Hemiptera: Pentatomidae). In Europe, the polyphagous Anastatus bifasciatus (Geoffroy) is the most prevalent native egg parasitoid on H. halys eggs and is currently being tested as a candidate for augmentative biological control. Anastatus bifasciatus frequently displays behavior without oviposition, and induces additional host mortality through oviposition damage and host feeding that is not measured with offspring emergence. This exacerbates accurate assessment of parasitism and host impact, which is crucial for efficacy evaluation as well as for pre‐ and post‐release risk assessment. To address this, a general Anastatus primer set amplifying a 318‐bp fragment within the barcoding region of the cytochrome oxidase I (COI) gene was developed. When challenged with DNA of three Anastatus species —A. bifasciatus, Anastatus japonicus Ashmead, and Anastatus sp.—, five scelionid parasitoid species that might be encountered in the same host environments and 11 pentatomid host species, only Anastatus DNA was successfully amplified. When applied to eggs of the target host, H. halys, and an exemplary non‐target host, Dendrolimus pini L. (Lepidoptera: Lasiocampidae), subjected to host feeding, no Anastatus amplicons were produced. Eggs of the two host species containing A. bifasciatus parasitoid stages, from 1‐h‐old eggs to pupae, and emerged eggs yielded Anastatus fragments. Confirmation of parasitoid presence with dissections and subsequent PCRs with the developed primer pair resulted in 95% success for 1‐h‐old parasitoid eggs. For both host species, field‐exposed sentinel emerged eggs stored dry for 6 months, 100% of the specimens produced Anastatus amplicons. This DNA‐based screening method can be used in combination with conventional methods to better interpret host‐parasitoid and parasitoid‐parasitoid interactions. It will help address ecological questions related to an environmentally friendly approach for the control of H. halys in invaded areas.     

Nicotine induces cell survival and chemoresistance by stimulating Mcl-1 phosphorylation and its interaction with Bak in lung cancer

Nicotine is a major carcinogen in cigarettes, which can enhance cell proliferation and metastasis and increase the chemoresistance of cancer cells. Our previous data found that nicotine promotes cell survival in lung cancer by affecting the expression of antiapoptotic protein Mcl-1, suggesting that the Mcl-1 may be a therapeutic target for patients with lung cancer. In this study, we found that the effects of drug resistance on nicotine-induced lung cancer cell lines were shown to influence the phosphorylation of Mcl-1. Moreover, nicotine induces Mcl-1 phosphorylation exclusively at the T163 site, which results in enhancement of the antiapoptotic activity of Mcl-1 and increased cell survival. Meanwhile, nicotine can reduce the sensitivity of H1299 cells to CDDP via enhancement of the binding of Mcl-1 to Bak, which inhibits the proapoptotic effect of Bak and ultimately leads to increased survival and drug resistance of lung cancer cells. Thus, nicotine-induced cell survival and chemoresistance may occur in a mechanism by stimulating Mcl-1 phosphorylation and its interaction with Bak, which may contribute to improving the efficacy of chemotherapy in the treatment of human lung cancer.     

CtIP promotes G2/M arrest in etoposide-treated HCT116 cells in a p53-independent manner

Acquired resistance to cytotoxic antineoplastic agents is a major clinical challenge in tumor therapy; however, the mechanisms involved are still poorly understood. In this study, we show that knockdown of CtIP, a corepressor of CtBP, promotes cell proliferation and alleviates G2/M phase arrest in etoposide (Eto)-treated HCT116 cells. Although the expression of p21 and growth arrest and DNA damage inducible α (GADD45a), which are important targets of p53, was downregulated in CtIP-deficient HCT116 cells, p53 deletion did not affect G2/M arrest after Eto treatment. In addition, the phosphorylation levels of Ser317 and Ser345 in Chk1 and of Ser216 in CDC25C were lower in CtIP-deficient HCT116 cells than in control cells after Eto treatment. Our results indicate that CtIP may enhance cell sensitivity to Eto by promoting G2/M phase arrest, mainly through the ATR-Chk1-CDC25C pathway rather than the p53-p21/GADD45a pathway. The expression of CtIP may be a useful biomarker for predicting the drug sensitivity of colorectal cancer cells.     

一种中华鳖结节病的病原菌分离鉴定及病理研究

【背景】在山东聊城市一中华鳖(Trionyx sinensis)养殖场暴发了结节病(Sarcoidosis),典型症状为内脏、四肢肌肉等部位出现结节;感染后期引起多重感染,所有内脏发臭腐烂。调查发现2017年该病传染率、死亡率较2016年均升高。目前国内尚未有中华鳖感染结节病的报道,其感染途径、病原体及防治方法均属未知。【目的】分析中华鳖结节病的病原,进行病理学研究,并为防控该病寻找解决方案。【方法】按照病害诊断方法,在排除寄生虫及病毒感染后,进行病原菌的分离及鉴定。同时通过病原菌的体外药敏试验进行防控药物筛选,并对该病的组织病理学进行研究。【结果】从病灶处分离到两种优势菌株DM1和DM2,体外回归感染试验表明DM1为中华鳖结节病病原菌。16S rRNA基因序列测定及生理生化鉴定显示,DM1为鼠伤寒沙门氏菌(Salmonella typhimurium)。药敏试验显示,该菌株对丁胺卡那、庆大霉素、头孢哌酮、环丙沙星等14种药物敏感,对呋喃唑酮和头孢氨苄表现中敏,对诺氟沙星、利福平、氨苄西林等18种药物表现耐药。病理切片显示结节最外层为纤维结缔组织形成的包膜,内部豆渣样物质为嗜酸性凝固性坏死。【结论】中华鳖结节病的病原菌为鼠伤寒沙门氏菌,养殖过程中可通过使用丁胺卡那、庆大霉素等药物进行疾病防控。     

四株杂交鳢致病性舒伯特气单胞菌特性及致病性比较

【背景】舒伯特气单胞菌为革兰氏阴性短杆菌,是一种人-畜-鱼共患的条件性致病菌,它能引起不同程度的人类疾病,近年来更是在水产养殖中被频繁报道。【目的】比较分析4株杂交鳢源舒伯特气单胞菌(WL-4、ZL-1、ZL-13、D075)菌株间生理生化特性、致病性以及对常用药物的敏感性差异,为水产舒伯特气单胞菌病的防治提供参考依据。【方法】使用全自动细菌生化鉴定仪检测菌株的生理生化特性;运用PCR扩增和测序比对菌株的gyrB、16S rRNA基因序列;采用改良平板法检测毒力因子表达,PCR技术检测菌株毒力基因和耐药基因的携带情况;通过人工注射感染分析4株舒伯特气单胞菌对杂交鳢的致病性差异;并采用微量二倍稀释法测试15种抗菌药物对菌株的最小抑菌浓度。【结果】以ATCC43700 (人源舒伯特气单胞菌)作为参考,4株菌株理化特性基本相同,gyrB和16SrRNA基因一致性分别为99.57%和100%,聚类分析聚为一簇;不同的舒伯特气单胞菌(ATCC43700、WL-4、ZL-1、ZL-13、D075)均具有溶血性、蛋白酶活性和脂肪酶活性;但人源菌株(ATCC43700)与杂交鳢源舒伯特气单胞菌携带的毒力基因存在差异,其中人源菌株携带了hlyA、ela、lip、ahal、act,而杂交鳢源菌株(WL-4、ZL-1、ZL-13、D075)携带hlyA、ela、lip、ahal、aer;4株杂交鳢源舒伯特气单胞菌对杂交鳢致病性存在差异,1.2×10~5CFU/mL浓度腹腔注射感染健康杂交鳢,WL-4、ZL-1、ZL-13、D075对杂交鳢的感染致死率依次为30%、40%、70%和80%;不同菌株(ATCC43700、WL-4、ZL-1、ZL-13、D075)的药物敏感性以及携带的耐药基因也存在差异。【结论】4株杂交鳢源舒伯特气单胞菌在致病性、药物敏感性等方面存在差异,研究结果有助于进一步开展舒伯特气单胞菌病发病机制和防控技术研究。     

XRCC4, which is inhibited by PFDA,regulates DNA damage repair and cell chemosensitivity

The mechanism of environmental pollution promoting gastric cancer incidence and difficulty of treatment is not fully understood. In the present article, perfluorodecanoic acid (PFDA), a common persistent environmental pollutant, was used to treat the gastric cell lines and mice to test its genotoxicity. The γ-H2AX immunoblot and plasmid fragment PCR results showed that PFDA had a promotion effect on the DNA double-strand breaks (DSBs) in human and mouse cells. Subsequent results showed that PFDA significantly altered the sensitivity of cells to chemotherapy. Microarray data showed that the expressions of some important DNA repair genes were changed. Further investigation discovered that PFDA inhibition of DNA repair was mediated by X-ray repair cross complementing 4 (XRCC4). The cells deficient in XRCC4 generally exhibited reduced proliferation and premature aging in culture; however, our results indicated that PFDA induced p53 inhibition rescued cells from the apoptosis that was triggered by nonhomologous end-joining (NHEJ) inactivation, and overexpression of p53 expression in PFDA-treated cells enhanced their apoptosis. Finally, T-cell specific factor 4 was suggested by the results as an upstream regulator of XRCC4. This article revealed for the first time that perfluorinated chemicals affect chemotherapeutic sensitivity and the NHEJ pathway, and p53 reduction rescues cells from death.     

Demographic factors affecting population growth in giant gartersnakes

Demographic models provide insight into which vital rates and life stages contribute most to population growth. Integral projection models (IPMs) offer flexibility in matching model structure to a species’ demography. For many rare species, data are lacking for key vital rates, and uncertainty might dissuade researchers from attempting to build a demographic model. We present work that highlights how the implications of uncertainties and unknowns can be explored by building and analyzing alternative models. We constructed IPMs for the threatened giant gartersnake (Thamnophis gigas) based on published studies to determine where management efforts could be targeted to have the greatest effect on population persistence and what unknowns remain for future research. Given uncertainty in the survival of snakes during their first year, and in the form of the size-survival relationship, we modeled a range of scenarios and evaluated where models agree about factors influencing population growth and where discrepancies exist. For most scenarios, the survival of large adult females had the greatest influence on population growth, but the relative importance of juvenile versus adult somatic growth for population growth was dependent on the recruitment probability and the shape of the size-survival function. More data on temporal variation and covariance among vital rates would improve stochastic models for the giant gartersnake. This paper demonstrates the effectiveness of IPMs for studying the demography of reptiles and the value of the model-building process for formalizing what is known and unknown about the demography of rare species. Published 2019. This article is a U.S. Government work and is in the public domain in the USA.     

Response of shrubland birds to regenerating clearcut area and shape

Conservation of birds that breed in early-successional forests will require an understanding of their response to patch size and shape during the early seral stages following disturbance. We modeled the effects of patch area, patch shape, and time period following harvest on territory densities of 5 shrubland bird species and bird community composition in 36 clearcuts from 1997 to 2014 in the Missouri Ozarks, USA. Our best-supported models indicated that densities of indigo bunting (Passerina cyanea), yellow-breasted chat (Icteria virens), and prairie warbler (Setophaga discolor) declined from the early time period (1–7 years post-harvest) to the late time period (12–18 years post-harvest), regardless of clearcut area or shape. Yellow-breasted chat density and bird species richness were positively related to clearcut shape complexity. Bird species richness increased, and bird species density decreased with clearcut area during both time periods. Non-metric multidimensional scaling indicated that bird community composition was strongly related to time since harvest. Within the spatial scale of our study, successional stage is more influential than clearcut size and shape, and some bird species may benefit from relatively small or irregularly shaped clearcuts. © 2019 The Wildlife Society.