An experiment of Bartonella henselae infection in Sprague-Dawley rats

doi:10.11853/j.issn.1003.8280.2021.06.002

Abstract

Abstract: Objective To infect Sprague-Dawley (SD) rats with Bartonella henselae strains and measure the dynamic changes of bacteremia and antibody in rats. Methods SD rats were infected by subcutaneous multi-point injection of different concentrations of B. henselae suspension. Whole blood was collected and diluted for culture. The heart, liver, spleen, and kidney were homogenized and isolated for culture. The probable B. henselae colony nucleic acids were extracted, and the citrate synthase gene (gltA) was amplified by PCR for sequence analysis. The antibody changes were measured by the indirect immunofluorescence antibody assay. SPSS 19.0 software was used to analyze the differences in the positive rate between different tissues, different suspension concentrations, and different days of infection using the Chi-square test and two-way analysis of variance (ANOVA). Results A total of 90 whole blood samples, 82 tissue samples, and 21 serum samples were collected, all of which were negative for culture. For whole blood samples, the positive rate of the nucleic acid was 30.56% (22/72), and positive samples were found on days 1, 3, and 5, with the highest positive rate of 100% occurring in the low-dose group and high-dose group on day 3. For tissue samples, the positive rate of the nucleic acid was 22.86% (16/70), and positive samples were found on days 3 and 5; the renal tissues showed the highest positive rate (7/16) and the most persistent positive status (lasting 5 days), and the liver tissues had the second highest positive rate (5/18). The antibody was present in 6 of 18 serum samples detected, beginning on the 7th day since infection till the 14th day when the experiment ended. No significant difference was observed in the positive rate between different dose groups (F=3.243, P=0.082, by two-way ANOVA) or between different types of samples (χ²=7.655, P=0.057, by the Fisher's exact test). The positive rate showed a significant difference between different times of infection (F=11.770, P=0.001, by two-way ANOVA), and specifically between the 3rd day and the 1st, 5th, 7th, 10th, or 14th day (all P<0.05). Conclusion B. henselae can infect SD rats for a short time, with nucleic acid positive rate peaking on the 3rd day of infection. The antibody can last at least two weeks since infection.

Key words: Bartonella henselae, Sprague-Dawley rat, Polymerase chain reaction, Indirect immunofluorescence antibody assay

摘要： 目的用汉赛巴尔通体菌株感染SD大鼠，检测大鼠体内菌血症及抗体的动态变化。方法用不同浓度的汉赛巴尔通体菌液经皮下多点注射感染SD大鼠，采集其全血稀释培养，心、肝、脾和肾脏组织匀浆后分离培养，提取疑似汉赛巴尔通体菌落核酸检测，用PCR扩增枸橼酸合酶基因（gltA），进行测序分析，间接免疫荧光抗体测定法测定抗体变化情况。应用SPSS 19.0软件进行统计分析，采用χ²检验和配伍组方差分析，分析不同感染浓度、不同组织间阳性率和不同感染天数的阳性率差异。结果共取样品全血90份、组织82份、血清21份，培养均为阴性。核酸检测全血阳性率为30.56%（22/72），其中第1、3、5天均有阳性样品，最高阳性率为100%（第3天的低剂量组和高剂量组），组织阳性率为22.86%（16/70），其中第3、5天均有阳性，阳性率最高的是肾脏，检测的16份样品中7份为阳性，其次是肝脏，检测的18份样品中5份为阳性，肾脏阳性率持续时间最长为5 d。检测血清18份，抗体阳性6份，最早出现在感染的第7天，持续到第14天实验结束。经配伍组方差分析，不同剂量组间阳性率差异无统计学意义（F=3.243，P=0.082）；经Fisher's确切概率法检验，不同组织样品阳性率差异无统计学意义（χ²=7.655，P=0.057）。经配伍组方差分析，不同感染天数阳性率差异有统计学意义（F=11.770，P=0.001），其中第3天与第1、5、7、10和14天的阳性率差异有统计学意义（均P<0.05）。结论 SD大鼠可短期感染汉赛巴尔通体，感染的高峰值出现在感染后的第3天，抗体至少可持续到感染后第2周。

关键词: 汉赛巴尔通体, SD大鼠, 聚合酶链式反应, 间接免疫荧光抗体测定法

CLC Number:

Q939.93

KANG Dong-mei, LI Dong-mei, SONG Xiu-ping, ZHANG Wen-zhu, LIU Qi-yong, LUN Xin-chang, MENG Feng-xia. An experiment of Bartonella henselae infection in Sprague-Dawley rats[J]. Chinese Journal of Vector Biology and Control, 2021, 32(6): 660-665.

康东梅, 栗冬梅, 宋秀平, 张文竹, 刘起勇, 伦辛畅, 孟凤霞. 汉赛巴尔通体感染SD大鼠实验研究[J]. 中国媒介生物学及控制杂志, 2021, 32(6): 660-665.

References

[1] Oray M,Önal S,Akbay AK,et al. Diverse clinical signs of ocular involvement in cat scratch disease[J]. Turk J Ophthalmol,2017,47(1):9-17. DOI:10.4274/tjo.28009.
[2] Sykes JE,Henn JB,Kasten RW,et al. Bartonella henselae infection in splenectomized domestic cats previously infected with hemotropic Mycoplasma species[J]. Vet Immunol Immunopathol,2007,116(1/2):104-108. DOI:10.1016/j.vetimm.2006.12.004.
[3] 杨小冉,刘起勇,崔步云,等. BALB/c及昆明小鼠的汉赛巴尔通体实验室感染模型[J]. 中国媒介生物学及控制杂志,2008,19(1):41-43. DOI:10.3969/j.issn.1003-4692.2008.01.015. Yang XR,Liu QY,Cui BY,et al. Characterization of infective model in Bartonella henselae-specific immunity in BALB/c and KM mice[J]. Chin J Vector Biol Control,2008,19(1):41-43. DOI:10.3969/j.issn.1003-4692.2008.01.015.
[4] Arvand M,Ignatius R,Regnath T,et al. Bartonella henselae-specific cell-mediated immune responses display a predominantly Th1 phenotype in experimentally infected C57BL/6 mice[J]. Infect Immun,2001,68(10):6427-6433. DOI:10. 1128/IAI.69.10.6427-6433.2001.
[5] Polar RC,Orellana G,Caso WS,et al. Encephalitis with convulsive status in an immunocompetent pediatric patient caused by Bartonella henselae[J]. Asian Pacific J Trop Med,2016,9(6):610-613. DOI:10.1016/j.apjtm.2016.03.030.
[6] Chiaraviglio L,Duong S,Brown DA,et al. An immunocompromised murine model of chronic Bartonella infection[J]. Am J Pathol,2010,176(6):2753-2763. DOI:10.2353/ajpath.2010.090862.
[7] 白瑛,Kosoy MY,Maupin GO,等. 首次证实巴尔通体在我国云南鼠群中流行[J]. 中国人兽共患病杂志,2002,18(3):5-9. DOI:10.3969/j.issn.1002-2694.2002.03.001. Bai Y,Kosoy MY,Maupin GO,et al. Discovery of Bartonella species in rodents in Yunnan[J]. Chin J Zoonoses,2002,18(3):5-9. DOI:10.3969/j.issn.1002-2694.2002.03.001.
[8] 宋秀平,刘起勇,鲁亮,等. 海南省小型兽类巴尔通体的分离培养和序列分析[J]. 中国媒介生物学及控制杂志,2010,21(2):131-133. Song XP,Liu QY,Lu L,et al. Isolation and sequence analysis of Bartonella in small mammals in Hainan province[J]. Chin J Vector Biol Control,2010,21(2):131-133.
[9] 叶曦,姚美琳,李国伟. 福建省鼠形动物巴尔通体感染调查[J]. 中国人兽共患病学报,2006,22(8):779-781. DOI:10.3969/j.issn.1002-2694.2006.08.024. Ye X,Yao ML,Li GW. The investigation on the infection of Bartonella species in rodent hosts in Fujian[J]. Chin J Zoonoses,2006,22(8):779-781. DOI:10.3969/j.issn.1002-2694.2006. 08.024.
[10] 陈秀英,雷永良,柳付明,等. 浙江省丽水市鼠类巴尔通体的检测研究[J]. 中国卫生检验杂志,2013,23(1):118-119. Chen XY,Lei YL,Liu FM,et al. Study on the detection of Bartonella of Lishui rodents in Zhejiang province[J]. Chin J Heal Inspect,2013,23(1):118-119.
[11] 赖瑞青,徐晓红,黄洁玲,等. 33例猫抓病性淋巴结炎超声图像分析[J]. 广东医学院学报,2010,28(1):45-47. DOI:10.3969/j.issn.1005-4057.2010.01.019. Lai RQ,Xu XH,Huang JL,et al. Ultrasonographic analysis of 33 cases of cat scratch disease lymphadenitis[J]. J Guangdong Med Coll,2010,28(1):45-47. DOI:10.3969/j.issn.1005-4057.2010. 01.019.
[12] 宋秀平,栗冬梅,贾丽军,等. 内蒙古小型兽类巴尔通体感染情况调查[J]. 中国媒介生物学及控制杂志,2015,26(3):233-237. DOI:10.11853/j.issn.1003.4692.2015.03.004. Song XP,Li DM,Jia LJ,et al. Investigation of Bartonella infection in small mammals in Inner Mongolia,China[J]. Chin J Vector Biol Control,2015,26(3):233-237. DOI:10.11853/j.issn.1003.4692.2015.03.004.
[13] 左双燕. 我国黑龙江林区鼠型动物巴尔通体感染调查与分离鉴定[D]. 长沙:中南大学,2012. Zuo SY. Detection and isolation of Bartonella spp. in rodents in Helongjiang province of China[D]. Changsha:Central South University,2012.
[14] 王廷兆. 喜忧参半"宠物热"[J]. 环境,2002,5(10):27-28. Wang TZ. Pet fever[J]. Environment,2002,5(10):27-28.
[15] 刘纳新. 猫抓病23例明确诊断与治疗分析[J]. 中国误诊学杂志,2010,10(9):2204-2205. Liu NX. Diagnosis and treatment of cat scratch disease:an analysis of 23 cases[J]. Chin J Misd,2010,10(9):2204-2205.
[16] Brouqui PP,Houpikian XY,Lepedi H,et al. Virulence-related factors of Bartonella:observation of Bartonella infection in vitro and in vivo[M]//Kazar J,Toman R. Rickettsiae and Rickettsial Diseases Veda. Bratislava:Slovakia, 1996:622-627.
[17] Greene CE,Mcdermott M,Jameson PH,et al. Bartonella henselae infection in cats:evaluation during primary infection,treatment,and rechallenge infection[J]. J Clin Microbiol,1996,34(7):1682-1685. DOI:10.1128/jcm.34.7.1682-1685.1996.
[18] 栗冬梅,徐爱玲,宋秀平,等. 巴尔通体在自然感染的啮齿动物组织中的分布[J]. 中国媒介生物学及控制杂志,2021,32(2):181-187. DOI:10.11853/j.issn.1003.8280.2021.02.012. Li DM,Xu AL,Song XP, et al. Bartonella distribution in naturally infected rodent tissues[J]. Chin J Vector Biol Control,2021,32(2):181-187. DOI:10.11853/j.issn.1003.8280.2021.02.012.
[19] 康东梅,师灿南,开文龙,等. 宿主动物对不同宿主型猫栉首蚤吸血率和饱血率的影响[J]. 中国媒介生物学及控制杂志,2017,28(3):197-200. DOI:10.11853/j.issn.1003.8280.2017. 03.001. Kang DM,Shi CN,Kai WL, et al. Effect of hosts on blood-sucking and blood engorging in different host type cat fleas,Ctenocephalides felis[J]. Chin J Vector Biol Control,2017,28(3):197-200. DOI:10.11853/j.issn.1003.8280.2017.03.001.