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Article(id=1307013638070489965, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307013423896753139, articleNumber=null, orderNo=null, doi=10.11843/j.issn.0366-6964.2026.03.051, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1750608000000, receivedDateStr=2025-06-23, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1789545934454, onlineDateStr=2026-09-16, pubDate=1774195200000, pubDateStr=2026-03-23, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1789545934454, onlineIssueDateStr=2026-09-16, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1789545934454, creator=13701087609, updateTime=1789545934454, updator=13701087609, issue=Issue{id=1307013423896753139, tenantId=1146029695717560320, journalId=1306274286668972124, year='2026', volume='57', issue='3', pageStart='1177', pageEnd='1776', issueExtLink='null', onlineDate='null', pubDate='1774195200000', pubDateStr='2026-03-23', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1789545883392, creator='13701087609', updateTime=1789609930841, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1307282058464743901, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307013423896753139, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1307282058464743902, tenantId=1146029695717560320, journalId=1306274286668972124, issueId=1307013423896753139, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=1772, endPage=1776, ext={EN=ArticleExt(id=1307013638301176686, articleId=1307013638070489965, tenantId=1146029695717560320, journalId=1306274286668972124, language=EN, title=Identification and Pathogenicity Analysis of Bordetella bronchiseptica Isolated from Canine, columnId=1307007462452777247, journalTitle=Acta Veterinaria et Zootechnica Sinica, columnName=RESEARCH NOTES, runingTitle=null, highlight=null, articleAbstract=

This study aimed to identify three suspected strains isolated from nasal swabs of dogs with upper respiratory tract infections at a police dog base in Shenyang. Species identification of isolated strains through 16S rRNA gene detection， phylogenetic tree construction. Pathogenicity analysis was conducted through virulence gene testing， drug susceptibility testing， and canine challenge test.The results showed that three strains isolated in this experiment are Bordetella bronchiseptica， named SY-202401， SY-202402 and SY-202403 respectively.Strains are resistant to 9 antibiotics， and sensitive to 6 antibiotics； After being infected with SY-202402 strain， the dogs showed symptoms such as coughing， retching， fervescence and increased eye and nose secretions，one dog die. The isolated Bordetella bronchiseptica from dogs in this study have high drug resistance and strong pathogenicity to dogs.

, authors=Fuxiao XIU1, Shibing ZHOU1, Pengpeng WANG1, Bingzhi LI2, Yuxiang YANG1, Ning SUN1, Min YANG1, Chengwu LIU1, authorsList=Fuxiao XIU, Shibing ZHOU, Pengpeng WANG, Bingzhi LI, Yuxiang YANG, Ning SUN, Min YANG, Chengwu LIU, authorCompany=null, correspAuthors=Shibing ZHOU, authorNote=null, correspAuthorsNote=
ZHOU Shibing， E-mail：
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旨在对沈阳某警犬基地患上呼吸道感染犬分离到的3株菌株进行鉴定和致病性分析。通过16S rRNA基因检测，系统进化树分析对分离菌株进行种属鉴定。通过毒力基因检测，药敏试验和犬攻毒试验进行致病性分析。结果显示：本试验分离到的3株菌株为支气管败血波氏杆菌（Bordetella bronchiseptica，Bb），分别命名为SY-202401、SY-202402和SY-202403；菌株对阿莫西林、氨苄西林、土霉素等9种抗生素耐药，对恩诺沙星、阿米卡星、链霉素等6种抗生素敏感；SY-202402株攻毒后，犬出现咳嗽、干呕、体温升高和眼鼻分泌物增多等症状，1头犬死亡。综上，本研究分离到的犬支气管败血波氏杆菌耐药性较高，对犬的致病性较强。

, authors=修福晓1, 周士兵1, 王朋朋1, 李炳志2, 杨宇翔1, 孙宁1, 杨敏1, 刘成武1, authorsList=修福晓, 周士兵, 王朋朋, 李炳志, 杨宇翔, 孙宁, 杨敏, 刘成武, authorCompany=null, correspAuthors=周士兵, authorNote=

修福晓，副研究员，硕士，主要从事动物病原学研究，E-mail：

, correspAuthorsNote=
周士兵，主要从事警犬技术教学与科研工作，E-mail：
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修福晓，副研究员，硕士，主要从事动物病原学研究，E-mail：

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修福晓，副研究员，硕士，主要从事动物病原学研究，E-mail：

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DOI:10.1128/mbio.00500-15, articleTitle=null, refAbstract=null), Reference(id=1307281945147236589, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013638070489965, doi=null, pmid=null, pmcid=null, year=null, volume=null, issue=null, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[16], rfOrder=18, authorNames=null, journalName=null, refType=null, unstructuredReference=CHEN Q，LEE G，CRAIG C，et al.A novel bvg-repressed promoter causes vrg-Like transcription of fim3 but does not result in the production of serotype 3 fimbriae in bvg-mode Bordetella pertussis ［J］.J Bacteriol，2018，24，200（20）：175-180. 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A. PCR result of 16S rRNA（M. DL2000 DNA marker；1.SY-202401； 2.SY-202402； 3.SY-202403； N.Negative control； P.Positive control）； B.PCR result of fla （M.DL2000 DNA marker； 1.SY-202401； 2.SY-202402； 3.SY-202403； N.Negative control； P.Positive control）

, figureFileSmall=wYUOHNEYWHw8PVd4rc2pgw==, figureFileBig=+z5QWGN7i3z9d/Lez/QdMg==, tableContent=null), ArticleFig(id=1307281939765944527, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013638070489965, language=CN, label=图1, caption=Bb 16S rRNA和fla基因PCR检测结果

A.16S rRNA PCR结果（M. DL2000 DNA相对分子质量标准； 1.SY-202401； 2.SY-202402； 3.SY-202403； N.阴性对照； P.阳性对照）；B. fla PCR检测结果（M. DL2000 DNA相对分子质量标准； 1.SY-202401； 2.SY-202402；3.SY-202403； N.阴性对照； P.阳性对照）

, figureFileSmall=wYUOHNEYWHw8PVd4rc2pgw==, figureFileBig=+z5QWGN7i3z9d/Lez/QdMg==, tableContent=null), ArticleFig(id=1307281940294426833, tenantId=1146029695717560320, journalId=1306274286668972124, articleId=1307013638070489965, language=EN, label=Fig.2, caption=Clinical symptoms and autopsy results of dead dog

A. Eye secretions increased in dog on the 6th day of infection； B. Dog experienced dry vomiting （arrow） on the 5th day of infection； C. Spleen enlargement； D. Tracheal wall congestion； E.Generalized necrosis of lung； F. Lung pathology show granulomatous pneumonia， increased number of multinucleated macrophages（arrow）

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A.犬感染第6天，眼分泌物增多；B.犬感染第5天出现干呕（箭头）；C.死亡犬脾脏肿大；D.气管壁淤血（箭头）；E.肺叶泛发性坏死（箭头）；F.肺脏病理结果显示肉芽肿性肺炎，可见多核巨噬细胞增多（箭头）

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犬源支气管败血波氏杆菌的分离鉴定及致病性分析
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修福晓 1 , 周士兵 1 , 王朋朋 1 , 李炳志 2 , 杨宇翔 1 , 孙宁 1 , 杨敏 1 , 刘成武 1
畜牧兽医学报 | 研究简报 2026,57(3): 1772-1776
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畜牧兽医学报 |研究简报 2026 , 57 (3) : 1772 -1776
犬源支气管败血波氏杆菌的分离鉴定及致病性分析
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修福晓1 , 周士兵1 , 王朋朋1, 李炳志2, 杨宇翔1, 孙宁1, 杨敏1, 刘成武1
作者信息
  • 1.中国刑事警察学院警犬技术学院，沈阳 110034
  • 2.杨凌职业技术学院动物工程学院，杨凌 712100
通讯作者:
周士兵，主要从事警犬技术教学与科研工作，E-mail：
作者简介:

修福晓，副研究员，硕士，主要从事动物病原学研究，E-mail：

Identification and Pathogenicity Analysis of Bordetella bronchiseptica Isolated from Canine
Fuxiao XIU1 , Shibing ZHOU1 , Pengpeng WANG1, Bingzhi LI2, Yuxiang YANG1, Ning SUN1, Min YANG1, Chengwu LIU1
Affiliations
  • 1.Criminal Investigation Police University of China，Shenyang 110034，China
  • 2.College of Animal Engineering，Yangling Vocational and Technical College，Yangling 712100，China
    • Corresponding Author:
    ZHOU Shibing， E-mail：
出版时间: 2026-03-23 doi: 10.11843/j.issn.0366-6964.2026.03.051
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摘要
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旨在对沈阳某警犬基地患上呼吸道感染犬分离到的3株菌株进行鉴定和致病性分析。通过16S rRNA基因检测，系统进化树分析对分离菌株进行种属鉴定。通过毒力基因检测，药敏试验和犬攻毒试验进行致病性分析。结果显示：本试验分离到的3株菌株为支气管败血波氏杆菌（Bordetella bronchiseptica，Bb），分别命名为SY-202401、SY-202402和SY-202403；菌株对阿莫西林、氨苄西林、土霉素等9种抗生素耐药，对恩诺沙星、阿米卡星、链霉素等6种抗生素敏感；SY-202402株攻毒后，犬出现咳嗽、干呕、体温升高和眼鼻分泌物增多等症状，1头犬死亡。综上，本研究分离到的犬支气管败血波氏杆菌耐药性较高，对犬的致病性较强。

支气管败血波氏杆菌  /  药敏试验  /  毒力基因

This study aimed to identify three suspected strains isolated from nasal swabs of dogs with upper respiratory tract infections at a police dog base in Shenyang. Species identification of isolated strains through 16S rRNA gene detection， phylogenetic tree construction. Pathogenicity analysis was conducted through virulence gene testing， drug susceptibility testing， and canine challenge test.The results showed that three strains isolated in this experiment are Bordetella bronchiseptica， named SY-202401， SY-202402 and SY-202403 respectively.Strains are resistant to 9 antibiotics， and sensitive to 6 antibiotics； After being infected with SY-202402 strain， the dogs showed symptoms such as coughing， retching， fervescence and increased eye and nose secretions，one dog die. The isolated Bordetella bronchiseptica from dogs in this study have high drug resistance and strong pathogenicity to dogs.

Bordetella bronchiseptica  /  drug sensitivity test  /  virulence gene
修福晓, 周士兵, 王朋朋, 李炳志, 杨宇翔, 孙宁, 杨敏, 刘成武. 犬源支气管败血波氏杆菌的分离鉴定及致病性分析. 畜牧兽医学报, 2026 , 57 (3) : 1772 -1776 . DOI: 10.11843/j.issn.0366-6964.2026.03.051
Fuxiao XIU, Shibing ZHOU, Pengpeng WANG, Bingzhi LI, Yuxiang YANG, Ning SUN, Min YANG, Chengwu LIU. Identification and Pathogenicity Analysis of Bordetella bronchiseptica Isolated from Canine[J]. Acta Veterinaria et Zootechnica Sinica, 2026 , 57 (3) : 1772 -1776 . DOI: 10.11843/j.issn.0366-6964.2026.03.051
正文
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支气管败血波氏杆菌（Bordetella bronchiseptica， Bb）属于β变形杆菌纲产碱杆菌科博特氏菌属，在自然界中广泛分布，主要引起犬、猪、猫和兔等多种哺乳动物呼吸道疾病［1］。Bb常与犬腺病毒、支原体、犬副流感、犬瘟热、犬疱疹病毒等中的一种或多种混合感染，从而引发犬传染性呼吸道疾病（canine infectious respiratory disease，CIRD），该病世界各地均有报道。近些年研究发现，人感染Bb病例逐渐增多［2］，表明Bb可对公共卫生安全造成一定威胁。本试验对沈阳某警犬基地分离到的犬源支气管败血波氏杆菌进行生物学特性分析，并研究其致病力，旨在为犬源支气管败血波氏杆菌病原学的研究及诊疗提供数据支持和理论依据。
1　材料与方法
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1.1　材料
1.1.1　样本来源
2024年4月和11月，沈阳某警犬基地患上呼吸道感染犬的鼻、咽拭子。
1.1.2　主要试剂与仪器
DL-2000 Marker、dNTP（0.5 mL·只-1）和Taq DNA聚合酶（250 IU·只-1），购自宝生生物工程有限公司；药敏纸片购自江苏维赛科技生物发展有限公司。PowerPac HC电泳仪、PCR仪、凝胶成像仪均购自美国Bio-Rad公司。
1.2　方法
1.2.1　细菌分离及革兰染色
采用无菌棉拭子采集患犬的鼻分泌物，经纯化培养后，挑取单一菌落涂抹于洁净载玻片上，进行革兰染色，油镜下对细菌进行染色特性观察。
1.2.2　分子鉴定及进化树分析
采用16S rRNA通用引物扩增分离菌的16S rRNA基因片段［3］。参照Akerley等［4］设计的引物对支气管败血波氏杆菌鞭毛基因（fla）进行PCR检测。对阳性产物进行测序并与NCBI 核酸序列数据库进行同源性比对。
1.2.3　生化反应鉴定
挑取单菌落分别接种于细菌微量发酵管进行如下生化试验：葡萄糖、蔗糖、乳糖、氧化/发酵（O/F）、肌醇、山梨醇、VP、MP、吲哚、枸橼酸钠、触酶、氧化酶、尿素和硝酸盐试验，根据不同生化反应时间要求定时观察。
1.2.4　药敏试验
采用纸片扩散法对3株菌株进行药敏试验。将菌液浓度调至3.0×108 CFU·mL-1，吸取菌液150 μL至TSA平板上涂布，选取8类共18种抗生素药敏纸片贴片。在37 ℃恒温培养箱中培养 48 h，测量抑菌圈直径，根据CLSI/NCCLS标准判断药物敏感性。
1.2.5　毒力基因检测
采用PCR检测Bb 5个主要毒力基因：细胞坏死毒素（dnt）、菌毛（fim）、黏附素（prn）、腺苷酸环化酶溶血素（cyaA）和III 型分泌系统效应物（bteA）。参考Wang等［5］进行引物设计。
1.2.6　犬的致病性试验
选取SY-202402菌株进行攻毒试验。SY-202402菌株经TSA液体培养基培养12 h后，用无菌 PBS 将菌量调整为5×109 CFU·mL-1，选取5头6~8月龄健康马里努阿犬（分别编号为A、B、C、D和E），对其滴鼻接种1 mL菌液，连续观察10 d试验犬的临床症状，并采集鼻拭子进行细菌的分离鉴定。
2　结果
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2.1　细菌的分离培养与革兰染色
从沈阳某警犬基地患上呼吸道感染犬只中分离得到3株疑似支气管败血波氏杆菌，分别命名为SY-202401、SY-202402和SY-202403。将疑似菌落纯化培养并进行革兰染色，油镜下为革兰阴性细菌，呈杆状或球状。
2.2　分子鉴定及遗传分析
采用PCR方法对3株菌株的16S rRNA基因和fla基因扩增，结果显示均扩增出约1 500和237 bp的目的条带（图1）。将16S rRNA基因的测序结果上传至NCBI进行序列比对，证实分离到的菌株均为支气管败血波氏杆菌。
2.3　药敏试验
分离株对阿莫西林、氨苄西林、阿奇霉素、红霉素、头孢呋锌、头孢曲松、磺胺嘧啶、磺胺甲噁唑、土霉素9种抗生素耐药，对环丙沙星、链霉素、恩诺沙星、阿米卡星、链霉素等6种抗生素敏感，对庆大霉素、卡那霉素和多西环素中度敏感。
2.4　生化试验
分离菌株不发酵各种糖类、不分解碳水化合物；甲基红（MR）、VP和吲哚反应均为阴性；分解尿素，氧化酶阳性。与波氏杆菌生化反应特性一致［6］。
2.5　毒力基因检测
SY-202401和SY-202403各检测到3种毒力基因（fim、prn和bteA），SY-202402株检测到4种毒力基因（fim、prn、dnt和bteA），所有菌株均未检测到cyaA基因。
2.6　犬的攻毒试验
5头犬攻毒后，均表现体温升高、鼻分泌物增多和咳嗽；有4头犬出现干呕，3头犬出现眼分泌物增多。编号D试验犬死亡。对死亡解剖发现死亡犬脾脏肿大，气管壁淤血，肺脏泛发性坏死，肺脏病理显示为肉芽肿性肺炎（图2）。攻毒后每日采集鼻拭子，按照“1.2.1”和“1.2.2”的方法进行鉴定，结果显示，从攻毒第3天开始，能够在鼻拭子分离检测到支气管败血波氏杆菌。
3　讨论
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支气管败血波氏杆菌可引发包括人、猪、犬、猫和兔等多种动物呼吸道感染。不同物种、不同地区Bb的流行情况存在一定差异。如：Rodriguez和Berliner［7］对美国16所猫舍的流行病学调查发现，Bb的感染率达到25%；Tabatabaei和Rohani［8］对伊朗63头犬咽拭子PCR检测，发现Bb阳性率达39.68%。Bb多是与一至多种病原体共同感染引发犬的支气管炎，如Jang等［9］对3 994篇文献采用meta回顾分析，发现Bb与其他病原体共同感染比例达到47%。这种多病原混合性感染模式增加了该病的防控难度。值得注意的是，犬和猫作为伴侣动物会将Bb传染给宠物主人或相关从业人员，如：Dworkin等［10］研究发现，9名被Bb感染患者（患有HIV）中，两名患者在生病前与犬有家庭接触，一名患者与猫有家庭接触；Clements等［11］研究发现，1名35岁与犬有家庭接触史的男性被Bb感染后，出现严重的呼吸系统疾病。因此，加强对犬、猫等伴侣动物的Bb流行病学调查具有重要的公共卫生意义。
细菌具有复杂的耐药机制，有学者针对Bb耐药基因进行了相关检测，以从遗传学角度进行耐药机制分析。不同学者针对不同来源Bb所分析得到的耐药基因存在一定差异，如Kadlec等［12］对猪源Bb耐药基因分析，发现猪源Bb携带甲氧苄啶抗性基因dfrA1和氯霉素抗性基因catB3。李雅卓等［13］对犬源Bb耐药基因分析发现：2个氨基糖苷类耐药基因和1个磺胺类耐药基因。需要注意的是，对耐药基因进行检测可以从遗传学角度更好地分析和研究细菌的耐药机制，但是耐药基因有无不等同于细菌的实际耐药性，如：赵梦溪［14］研究发现，猪源所有Bb分离菌株均对β-内酰胺类和部分氨基糖苷类药物具有耐药性，但均未检测出相关耐药基因。临床上在治疗支气管败血波氏杆菌引发的疾病，应尽可能的先进行药敏试验，并依据药敏试验结果选择合适的抗生素进行治疗。
所有鲍特氏菌属细菌感染机体的先决条件是能够在呼吸道定殖。而毒力因子在细菌呼吸道定殖中起到关键作用。本研究对分离到3株Bb毒力基因检测表明，所有菌株均存在fim、prn和bteA基因。有研究表明，毒力因子的表达受bvg （Bordetella virulencegenes）基因座的协同调节，在bvg基因座调节下表达的毒力因子主要包括腺苷酸环化酶溶血素、黏附素和菌毛［15］。综上，fim、prn和bteA基因在鲍特氏菌属细菌呼吸道定殖和感染中起到重要作用。在鲍特氏菌属中，上述基因虽然存在，但并非所有细菌都表达。如，fimX亚基基因虽然存在于百日咳杆菌中，但该基因是沉默的［16］。因此，致病性不仅依赖毒力基因的存在，还受调控机制的影响。
本研究虽然在一定程度上填补了支气管败血波氏杆菌部分研究空白，但仍存在一定的局限性，如：样本量小，样本采集时间跨度短。为更好地对支气管败血波氏杆菌进行系统性和延续性的分析，需要对来自不同地区和时间的更多样本，持续性进行呼吸道病原调查和分析，为犬呼吸道疾病的病原诊断和预防治疗提供数据支持。
4　结论
收起
本试验成功分离并鉴定3株犬源支气管败血波氏杆菌，检测了上述菌株的耐药性，可为临床用药提供参考依据。对相关菌株毒力基因和致病性进行了分析，为犬支气管败血波氏杆菌的疫病诊断及防控方向研究提供了科学依据。
基金
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  • 国家重点研发计划项目(2017YFC0820301)
  • 中国刑事警察学院重点攻关项目(D2025013)
  • 中国刑事警察学院周士兵专家工作室专项建设经费（30299）
文献
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参考文献 引证文献
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2026年第57卷第3期
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doi: 10.11843/j.issn.0366-6964.2026.03.051
  • 接收时间：2025-06-23
  • 首发时间：2026-09-16
  • 出版时间：2026-03-23
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  • 收稿日期：2025-06-23
基金
国家重点研发计划项目(2017YFC0820301)
中国刑事警察学院重点攻关项目(D2025013)
中国刑事警察学院周士兵专家工作室专项建设经费（30299）
作者信息
    1.中国刑事警察学院警犬技术学院，沈阳 110034
    2.杨凌职业技术学院动物工程学院，杨凌 712100

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周士兵，主要从事警犬技术教学与科研工作，E-mail：
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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