青海高原禾草内生真菌资源的调查和分离

青海高原的高寒特性决定其生境内禾草植物的多样性和特异性,然而有关青海高原地区禾草内生真菌的研究报道较少,本研究对青海高原地区48个样点内禾草植物内生真菌带菌率和内生真菌分离进行了研究.结果表明,14种禾本科植物内生真菌平均感染率低于50％,而12种植物内生真菌平均感染率大于50％.其中,缘毛鹅观草(Roegneria pendulina)、紫花针茅(Stipa purpurea)、醉马草(Achnatherum inebrians)、中华羊茅(Festuca sinensis)、硬质早熟禾(Poa sphondylodes)、微药羊茅(F.nitidula)、毛稃羊茅(F.kiritowii)和紫羊茅(F.rubra) 的平均带菌率均超过80％.同时,同属禾草植物内生真菌带菌率存在差异.禾草内生真菌分离率较低,其中紫花针茅、麦宾草(El ymus tangutorum)、光稃早熟禾(P.psiolepis)和异针茅(S.aliena)内生真菌在菌落和分生孢子形态上存在差异.可见,青海高原禾草内生真菌带菌率较高,同时不同禾草内生真菌可能存在多样性.     

Genetic characterization of bovine viral diarrhea virus strains in Beijing,China and innate immune responses of peripheral blood mononuclear cells in persistently infected dairy cattle

To acquire epidemiological data on the bovine viral diarrhea virus (BVDV) and identify cattle persistently infected (PI) with this virus, 4,327 samples from Holstein dairy cows were screened over a four-year period in Beijing, China. Eighteen BVD viruses were isolated, 12 from PI cattle. Based on genetic analysis of their 5''-untranslated region (5''-UTR), the 18 isolates were assigned to subgenotype BVDV-1m, 1a, 1d, 1q, and 1b. To investigate the innate immune responses in the peripheral-blood mononuclear cells of PI cattle, the expression of Toll-like receptors (TLRs), RIG-I-like receptors, interferon-α (IFN-α), IFN-β, myxovirus (influenza virus) resistance 1 (MX1), and interferon stimulatory gene 15 (ISG15) was assessed by qPCR. When compared with healthy cattle, the expression of TLR-7, IFN-α, and IFN-β mRNA was downregulated, but the expression of MX1 and ISG-15 mRNA was upregulated in PI cattle. Immunoblotting analysis revealed that the expression of interferon regulatory factor 3 (IRF-3) and IRF-7 was lower in PI cattle than in healthy cattle. Thus, BVDV-1m and 1a are the predominant subgenotypes in the Beijing region, and the strains are highly divergent. Our findings also suggest that the TLR-7/IRF-7 signaling pathway plays a role in evasion of host restriction by BVDV.     

Investigation of biofilm production and icaA and icaD genes in Staphylococcus aureus isolated from heifers and cows with mastitis

Biofilm formation and antimicrobial resistance of Staphylococcus aureus are important virulence factors in cases of mastitis in dairy cows. However, few studies have investigated mastitis strains isolated from heifers. Within this context, the objective of the present study was to investigate biofilm formation on Congo red agar, the presence of the icaA and icaD genes by polymerase chain reaction (PCR), and the percentage of in vitro antimicrobial resistance of 110 S. aureus isolates from mammary gland secretions of heifers and cows with mastitis. PCR detected the icaA and icaD genes in 98% and 100% of isolates, respectively. However, only 55.5% of all isolates produced a biofilm on Congo red agar. Antimicrobial susceptibility testing revealed that 47.0% of isolates from heifers and 70.4% of isolates from cows were resistant to at least one of the antimicrobial agents tested. Resistance to penicillin and/or ampicillin was the most frequent (44.5%). These results indicate the need to implement prophylactic and control measures of mastitis for heifers. Heifers and cows can carry resistant strains with the capacity of biofilm production, a fact representing a threat to public health and animal well‐being and generating losses to dairy farmers.     

贵州某野生动物园长臂猿流感病毒、支原体及巴氏杆菌混合感染的诊治

为弄清贵州某野生动物园长臂猿死亡的原因,本试验采用流行病学调查、临床症状观察、病理剖检诊断和RT-PCR检测确诊等方法,对该野生动物园发病长臂猿进行了诊断。试验结果显示,流行病学调查、剖检病理初步诊断该野生动物园长臂猿疑似流感病毒、支原体和细菌混合感染,RT-PCR/PCR检测确诊发病长臂猿为流感病毒、支原体混合感染,细菌分离培养、生化特性鉴定和动物致病性试验确诊为多杀性巴氏杆菌感染。结果表明造成该野生动物园长臂猿发病死亡的原因为流感病毒、支原体和多杀性巴氏杆菌混合感染。根据诊断及药敏试验结果,制定出了免疫预防及治疗措施。     

Isolation,Identification and Multidrug-resistant Analysis of the New Sequence Type ST989 Strain of Enterococcus faecium

Gram-positive cocci were isolated from half a month diseased lambs with a fatal infection in a sheep farm in Minhe county, Qinghai province.The clinical symptoms observed were drooping, reduced feed intake, emaciated and fever.The infected lambs died several hours after the disease been observed.Necropsy of two dying lambs revealed that for one lamb, the liver was enlarged and capsule of the liver dropped, and other organs had no obvious changes;For the other one, the organs had no obvious changes.After gram staining, gram-positive cocci were observed from all the samples.Partial Tuf gene sequences of these isolates were amplified and sequenced for species identification, BLASTn conparison result revealed that they had the highest sequence similarity with that of Enterococcus faecium (E.faecium) AUS0085 (99.8%), we thus primarily confirmed that these strains belonged to E.faecium.Antimicrobial susceptibility test indicated that these strains could be divided into two classes.Class 1 was resistant to β-lactams, aminoglycosides, macrolides, tetracyclines, quinolones, rifamycins, lincomycin and clindamycin;Susceptible to chloromycetin and vancomycin.Class 2 was resistant to β-lactams, aminoglycosides, tetracyclines, quinolones, rifamycins;Intermediate resistance to erythromycin and roxithromycin;But susceptible to clarithromycin, azithromycin, clindamycin, lincomycin, chloromycetin and vancomycin.Multilocus sequence typing (MLST) based on seven housekeeping genes for E.faecium revealed that E.faecium lby1 strain in class 1 and E.faecium lbg3 strain in class 2 were defined as a new sequence type ST989.Phylogenetic tree analysis revealed the representative strain of these isolates, E.faecium lbg3, was clustered together with E.faecium ST468.The pathogenicity test revealed that E.faecium lby1 and lbg3 strains both had pathogenicity.     

Pathogen Analysis of Bovine Respiratory Disease Complex

To investigate the pathogen of bovine respiratory disease complex (BRDC) of a dairy farm in Guangxi, a strain of Mycoplasma and a strain of gram-negative pathogenic bacterium were isolated and identified by the means of field surveys, clinic observation, pathological examination, isolation studies and so on.Treatments were taken according to drug sensitivity test results.The Mycoplasma strain, growing on PPLO medium, formed typical "fried egg" colonies.A 448 bp of oppF fragment was amplified by PCR from the strain and had 98.4% nucleotide identity with Mycoplasma bovis reference isolate PG5 of USA.The biochemical features of the gram-negative bacterial isolate were same with Serratia marcescens.The PCR amplified 16S rDNA of the gram-negative pathogenic bacterium strain was 1 400 bp.It shared 99.0% nucleotide identity with other Serratia marcescens reference strains obtained from GenBank.Animal experiment showed that the gram-negative pathogenic bacterium isolate could cause the mice to die.The drug sensitivity tests showed all isolates were sensitive to spectinomycin, azithromycin, amikacin, gentamicin and neomycin.It was effective to treat with dexamethasone and spectinomycin.Pathogen analysis and drug treatment showed that the BRDC was caused by Mycoplasma bovis and Serratia marcescens.     

Transmission ecology of rodent‐borne diseases: New frontiers

Rodents are recognized reservoir hosts for many human zoonotic pathogens. The current trends resulting from anthropocene defaunation suggest that in the future they, along with other small mammals, are likely to become the dominant mammals in almost all human‐modified environments. Recent intricate studies on bat‐borne emerging diseases have highlighted that many gaps exist in our understanding of the zoonotic transmission of rodent‐borne pathogens. This has emphasized the need for scientists interested in rodent‐borne diseases to integrate rodent ecology into their analysis of rodent‐borne pathogen transmission in order to identify in more detail the mechanisms of spillover and chains of transmission. Further studies are required to better understand the true impact of rodent abundance and the importance of pathogen sharing and circulation in multi‐host– multi‐pathogen communities. We also need to explore in more depth the roles of generalist and abundant species as the potential links between pathogen‐sharing, co‐infections and disease transmission.     

青蒿中番茄斑萎病毒和烟草花叶病毒的分子鉴定及相关序列分析

番茄斑萎病毒(tomato spotted wilt virus, TSWV)和烟草花叶病毒(tobacco mosaic virus, TMV)是2种重要的植物病原病毒, 对多种经济作物的产量和品质均造成严重影响。2021年-2022年, 在云南省丽江市烟草种植区不同烟区采集叶片黄化、皱缩以及无症状的青蒿Artemisia caruifolia样品共计14份, 利用免疫金标速测卡和RT-PCR对其病原病毒进行检测。利用免疫金标速测卡检测结果显示, 在所检样品中有9份样品检测出TSWV, 检出率为64.28%, 有3份样品检测出TMV, 检出率为21.43%, 2种病毒复合侵染的检出率同样为21.43%;利用RT-PCR对复合侵染的3份样品进行分子检测, 结果显示, 在3份复合侵染青蒿样品中获得3条TSWV N基因序列、3条TMV cp基因序列和2条TMV RdRp部分序列。TSWV青蒿分离物与分离自云南的TSWV-2分离物相似性最高, 为99.6%;TMV青蒿分离物与分离自辽宁的TMV-Shenyang分离物和分离自云南的TMV-Yongren-1相似性最高, 均大于99.4%。这是首次发现TSWV和TMV 2种不同属病毒复合侵染青蒿。     

渝西地区兔球虫病流行情况调查及虫种鉴定

【目的】掌握渝西地区兔球虫病的流行情况和兔球虫种类,为该地区兔球虫病的防治提供依据。 【方法】采用饱和食盐水漂浮法和麦克马斯特氏计数法检测渝西荣昌、永川、大足等 8 个区 14 个兔场共 1 273 份 粪样本,并收集卵囊,鉴定兔球虫种类。【结果】发现 14 个兔场均有感染球虫,平均感染率为 90.65%,璧山 和江津的感染率最高,为 100%,2 月龄以下兔感染率极显著高于 2~4 月龄、4~9 月龄及 9 月龄以上年龄段兔, 感染率为 98.19%。小型养殖场兔球虫感染率极显著低于大型规模化养殖场,感染率为 86.53%。8 个区的克粪便 虫卵数（Oocysts Per Gram,OPG）为 9.65×103 ~6.56×104 ,其中 2~4 月龄 OPG 最高、为 6.31×104 。对药物使 用情况调查显示,未使用球虫药养殖场的感染率为 92.31%,OPG 为 4.42×104 ; 使用球虫药养殖场的感染率为 89.19%,OPG 为 2.75×104 。在 14 个兔场共鉴定了 10 种艾美耳球虫,其中优势虫种为穿孔艾美耳球虫（Eimeria perforans）、大型艾美耳球虫（E. magna）、盲肠艾美耳球虫（E. coecicola）和中型艾美耳球虫（E. media）, 检出率分别为 25.20%、23.38%、11.96%、10.02%。【结论】渝西地区普遍感染兔球虫,且多为混合感染,因此 应加强该地区兔球虫病的防治。