Genetic and ecologic characteristics of Bartonella communities in rodents in southern China

Ecologic and bacteriologic observations of small mammals captured in Yunnan Province in the People's Republic of China indicated that Bartonella infections occurred at a high prevalence among some rodent species. Sequence analyses of the citrate synthase genes of these Bartonella demonstrated that rodents in this region harbored a diverse assemblage of strains. The Bartonella isolates obtained from Apodemus, Eothenomys, and Rattus typically clustered separately by genus of rodent host. Cultures obtained from Rattus rats were genetically related to Bartonella elizabethae, a recognized human pathogen. The finding of Bartonella species in a high proportion of the rodent samples from Yunnan suggests the need to investigate whether these agents might be responsible for cases of febrile illnesses of unknown etiology in southern China and elsewhere in southeastern Asia.     

Prevalence of Bartonella species and 16s rRNA gene types of Bartonella henselae from domestic cats in Thailand.

Prevalence of Bartonella infection among 275 cats in 9 sites from 4 geographical regions (northern area: Chiang Mai; central area: Kanchanaburi, Ratchaburi, and Bangkok; northeastern area: Khon Kaen, Roi Et, Ubon Ratcharthani, and Nakhonratchasima; southern area: Songkhla) of Thailand was investigated. Overall, Bartonella species were isolated from 27.6% (76 of 275) of the cats. The isolation rate varied from 12.8% (5 of 39) in Songkhla (southern area) to 50.0% (26 of 52) in Khon Kaen (northeastern area). Bartonella henselae and B. clarridgeiae were isolated from 82.9% (63 of 76) and 11.8% (9 of 76) of the Bartonella-positive cats, respectively. Coinfection with both species was found in 5.3% (4 of 76) of the bacteremic cats. Of the 67 bacteremic cats from which B. henselae was isolated, 48 (71.6%) and 13 (19.4%) were infected with only Type I and Type II, respectively. Coinfection with both types was observed in 9.0% (6 of 67) of the B. henselae-positive cats. To our knowledge, this is the first report on the presence of Bartonella infection in domestic cats from Thailand, which constitute a large reservoir of Bartonella infection in this country.     

Occurrence of ectoparasites on rodents in Sukhothai Province, northern Thailand

A survey of ectoparasites on rodents was carried out bimonthly from April 2008 to March 2009 in 3 districts of Sukhothai Province, northern Thailand. A total of 130 rodents comprising 8 species of hosts were captured and examined for ectoparasites. The hosts examined were Bandicota indica, Bandicota savilei, Rattus losea, Rattus rattus, Rattus exulans, Rattus norvegicus, Menetes berdmorei and Tamiops mcclellandii. Ninety-seven ectoparasites were collected: 1 species of tick (Hemaphysalis bandicota), 2 species of mites (Laelaps nuttali and Laelaps echidninus), and 1 species of flea (Xenopsylla cheopis) were identified. The infestation rates by ticks, mites and fleas on the rodents were 0.77, 5.38 and 6.15%, respectively. Monitoring the rodent population and their ectoparasites is important for future planning of prevention and control of zoonotic diseases in the area.     

Different clinical features in patients with ankylosing spondylitis from southern and northern China

Aims: To describe clinical features of patients with ankylosing spondylitis (AS) from southern and northern China, and investigate the effects of onset age, gender and regional differences on disease phenotype. Methods: Totally 113 AS patients from southern China and 121 AS patients from northern China were analyzed retrospectively. Results: In southern and northern groups, low back pain was more frequent among initial symptoms (54.9% vs. 7.7%; 52.4% vs. 11.4%), while peripheral arthritis (15.7% vs. 35.9%; 22.2% vs. 68.6%) was less common in male adult AS (AAS) than in male juvenile AS (JAS) patients, respectively. Compared to those in the northern group, diagnostic delay was longer (7.3 vs. 3.5 years) and the prevalence of human leukocyte antigen (HLA)‐B27 was higher in the southern group (96.5% vs. 83.5%). Sacroiliitis grade 2 was more frequent (51.3% vs. 36.4%), while sacroiliitis grade 3 (32.7% vs. 53.7%), buttock pain (5.3% vs. 13.2%), knee (20.4% vs. 33.1%) and ankle (3.5% vs. 11.6%) arthritis were less frequent in the southern group. Diagnostic delay of southern JAS was longer than that of northern JAS regardless of gender. Both sacroiliitis grade 3 and peripheral arthritis were less frequent in southern male JAS than in northern male JAS. Diagnostic delay was longer, sacroiliitis grade 2 was more frequent, while sacroiliitis grade 3 was less frequent in southern male AAS than those in northern male AAS. Conclusion: Significant diagnostic delay and higher prevalence of HLA‐B27 were found in southern AS patients. The prevalence of buttock pain and peripheral arthritis at disease onset in northern AS was more frequent than in southern AS patients.     

Bartonella henselae: first reported isolate in a human in Thailand

This is the first reported case of bacillary angiomatosis associated with Bartonella henselae in Thailand. The clinical, pathological, and microbiological findings are presented. The bacterium was isolated from a biopsy of skin lesions obtained on admission and identified by cellular morphology, characteristics of colonies on chocolate agar, extensive biochemical tests and 16S ribosomal DNA sequencing.     

广东省鼠形动物携带巴尔通体的调查和基因特征分析

目的了解巴尔通体在广东省鼠形动物中的流行分布情况及其基因型特征,为巴尔通体的研究和自然疫源性疾病的防控提供科学依据。方法在广东省惠来县、惠东县、潮安县、罗定县和高州市5个地区进行捕鼠,采集鼠肝脏标本提取总核酸,利用聚合酶连反应(PCR)检测巴尔通体病原,并对其进行分离培养,阳性产物进行测序,并根据序列进行系统进化分析,统计分析巴尔通体的分布情况和特征。结果在广东省5个地区共捕鼠375只,检出巴尔通体阳性标本73份,阳性率为19.47%。不同鼠种(χ~2=6.361)、不同地区(χ~2=7.778)、不同性别(χ~2=0.292)、不同生境间(χ~2=0.621),阳性率差异均没有统计学意义(P>0.05)。73份样本中,10份成功分离培养出巴尔通体。系统进化分析表明广东鼠形动物携带的巴尔通体存在6种基因型:Bartonella elizabethae、B.phoceensis、B.japonica、B.henselae、B.rochalimae、B.tribocorum。结论巴尔通体在广东省鼠形动物中广泛存在,且基因型呈多样性,其中4种基因型对人类有致病性,对人类健康具有潜在威胁。     

Molecular screening of Bartonella species in rodents from south western Spain

Rodents (87 Mus spretus and 10 Rattus norvegicus) collected in a suburban area of Seville (Andalusia, Spain) between September 2003 and April 2004 were tested by polymerase chain reaction for the presence of Bartonella DNA using primers amplifying a fragment of the 16S/23S rRNA internal transcribed spacer (ITS). Sequence analyses of the ITS of these Bartonella demonstrated that rodents in this region harbored at least three strains: two Bartonella genotypes detected in M. spretus, which clustered separately, and one genotype corresponding with B. tribocorum in R. norvegicus. The finding of Bartonella species in a high proportion of the rodent samples suggests the need to investigate whether these agents might be responsible for cases of febrile illnesses of unknown etiology in Southern Spain.     

福建省鼠类感染巴尔通体调查及序列分析

目的了解福建省鼠类中巴尔通体(Bartonellaspp.)的感染状况和基因特征。方法 2014-2016年采用笼日法在福建省闽东、闽西、闽南、闽北和闽中捕鼠,现场鉴定并记录捕获鼠类的釆集时间、地点、鼠种、性别、鼠龄等资料。采集鼠心脏血,PCR扩增巴尔通体的gltA和16S~23SrRNA基因,阳性PCR产物送测序并进行序列比对分析,构建系统进化树。感染率间的比较采用χ~2检验或Fisher精确检验法。结果调查共布放鼠笼5 917笼次,捕鼠381只,鼠密度为6.44%。巴尔通体感染率为12.34%。家鼠的巴尔通体感染率为10.61%,褐家鼠(Rattus norvebicus)和黄胸鼠(Rattus flavipectus)感染率分别为11.30%和10.00%;野鼠的感染率为13.86%,黄毛鼠(Rattus losea)和针毛鼠(Rattus fulvescens)感染率分别为22.86%和18.00%,野鼠的巴尔通体感染率高于家鼠的感染率,但无统计学意义。从地区分布看,闽西、闽北一带的感染率较高,分别为20.00%和25.33%。闽南一带感染率最低,为0。各地区感染率存在统计学意义(P0.05)。不同性别、鼠龄的巴尔通体感染率差异无统计学意义,而不同的生境下的感染率存在统计学差异(P0.05)。阳性样本测序分析显示,福建省鼠类感染的巴尔通体序列与B.tribocorum、B.elizabethae和B.grahamii序列最接近。结论福建省鼠类存在巴尔通体感染,存在对人群致病的风险。     

首次证实巴尔通体在我国云南鼠群中流行

目的　了解巴尔通体 (Bartonella)在云南鼠群中的分布及流行特征。方法　被检鼠血为 1999年 10月收集自云南省的 3个调查地区 ,采用兔血心浸液琼脂培养基进行巴尔通体分离 ,以聚合酶链反应 (PCR)对枸橼酸合酶基因 (gltA)的379bp片段进行扩增以证实是否巴尔通体 ,阳性者行以序列测定并与已知菌株加以比较。 结果与结论　从 131份鼠血分离到5 8株巴尔通体 (44 3% )。菌株分布于各调查点 ,感染鼠分属 3个属 6个种 ,以姬鼠属 (Apodemus)的带菌率最高 (6 2 2 % ,2 8/4 5 ) ,家鼠属 (Rattus)次之 (41 5 % ,2 7/ 6 5 ) ,绒鼠属 (Eothenomys)居第三位 (18 8% ,3/ 16 ) ,表明巴尔通体在云南常见鼠种中广泛分布及高度流行。所有菌株按其分离鼠属可分为 3群 ,具有以属为水平的宿主特异性。依基因结构可将它们分为 2 0个变异体 (家鼠属 8个 ;姬鼠属 12个 ;绒鼠属 2个 ) ,其中 17个为新发现变异体 ,表明云南巴尔通体基因型别的多样性。 7个家鼠巴尔通体变异体可分为B elizabethae、B tribocorum和新种B yunnannensis 3个基因型。由于云南巴尔通体的高度流行及基因型别的多样性 ,一些不明原因的疾病可能与巴尔通体的感染有关。需要对该地区巴尔通体的流行情况及对人类致病作用进行系统的调查和研究     

浙江省鼠形动物巴尔通体的遗传进化分析

目的分析浙江省鼠形动物中巴尔通体分子遗传进化关系,为巴尔通体人群感染的预防控制提供科学依据。方法用夹夜法在浙江省不同地区、不同季节捕获鼠形动物,无菌操作取鼠肝和脾,用PCR和分离培养检测巴尔通体,对部分阳性产物测序,提交到GenBank,用CLUSTAL W进行匹配,然后用PAUP4.0beta10软件构建系统关系,分析其遗传进化关系。结果我们分别从黑线姬鼠、黄毛鼠、褐家鼠、黑腹绒鼠、社鼠、臭鼩鼱、东方田鼠、黄胸鼠和大林姬鼠中检测到巴尔通体特异DNA片段,浙江首次从黑线姬鼠脾中分离出一株巴尔通体。遗传进化分析显示我们检测到的巴尔通体与Bartonellaratti massiliensis以及对人类有致病性的B.grahamii的遗传关系最近。结论浙江省鼠类中广泛存在巴尔通体感染,而且携带人类致病性巴尔通体,存在人群感染风险。     

Subclinical atherosclerosis in northern and southern China: the Chinese paradox

Background The incidence of coronary heart disease (CHD) is higher in Northern than that in Southern China, however differences in traditional CHD risk factors do not fully explain this. No study has examined the differences in subclinical atherosclerosis that may help explain the differences in incidence. This study examined these differences in subclinical atherosclerosis using coronary computed tomography (CT) for calcification between the Northern and Southern China. Methods We selected a random sample of participants in a large multi-center ongoing epidemiologic study for coronary calcium scanning in one northern city (North) (Beijing, n = 49) and in two southern cities (South) (Shanghai, n = 50, and Guangzhou, n = 50). Participants from the three field centers (mean age 67 years) underwent coronary risk factor evaluation and cardiac CT scanning for coronary calcium measurement using the Multi-Ethnic Study of Atherosclerosis scanning protocol. Results Adjusted log-transformed coronary artery calcium score in North China (Beijing) was 3.1±0.4 and in South China (Shanghai and Guangzhou) was 2.2±0.3 (P = 0.04). Mean calcium score for the northern city of Beijing was three times higher than that of the southern city of Guangzhou (P = 0.01) and 2.5 times higher than for the southern city of Shanghai (P = 0.03). Conclusions The extent of subclinical atherosclerosis is significantly higher in the northern city of Beijing than that in the two southern cities of Guangzhou and Shanghai, even after adjusting for standard cardiac risk factors. This finding suggests that standard risk factors do not fully explain north south differences in clinical CHD incidence.     

Molecular Detection and Identification of Bartonella Species in Rat Fleas from Northeastern Thailand

The presence of Bartonella species in Xenopsylla cheopis fleas collected from Rattus spp. (R. exulans, R. norvegicus, and R. rattus) in Khon Kaen Province, Thailand was investigated. One hundred ninety-three fleas obtained from 62 rats, were screened by polymerase chain reaction using primers specific for the 16S–23S intergenic spacer region, and the presence of Bartonella DNA was confirmed by using the citrate synthase gene. Bartonella DNA was detected in 59.1% (114 of 193) of fleas examined. Sequencing demonstrated the presence of Bartonella spp. similar to B. elizabethae, B. rattimassiliensis, B. rochalimae, and B. tribocorum in the samples tested with a cutoff for sequence similarity ≥ 96% and 4 clustered together with the closest match with B. grahamii (95.5% identity). If X. cheopis proves to be a competent vector of these species, our results suggest that humans and animals residing in this area may be at risk for infection by several zoonotic Bartonella species.Bartonella species are small, pleomorphic, gram-negative bacteria that infect a variety of mammalian hosts, including cats, dogs, rodents, ruminants, and humans. Clinical symptoms associated with Bartonella range from mild, influenza-like symptoms to more severe manifestations such as endocarditis, myocarditis, uveitis, bacillary angiomatosis, and peliosis hepatis.¹ Approximately half of the 20 Bartonella species or subspecies identified to date are known or suspected human pathogens,² and most are believed to be transmitted by arthropod vectors (fleas, lice, sandflies, and ticks).³Xenopsylla cheopis, the Oriental rat flea, is a suspected vector of several Bartonella species (B. tribocorum, B. elizabethae, B. queenslandensis, B. rochalimae, and novel Bartonella genotypes), and Bartonella DNA has been detected in these fleas from various locations worldwide.^3–8 Although generally found on rodents, X. cheopis have been found to parasitize humans and are known vectors of the zoonotic agents Yersinia pestis (plague) and Rickettsia typhi (murine typhus).⁹Numerous surveys have been performed to identify the presence of Bartonella species affecting humans and domestic and peri-domestic animals in Thailand.^10–17 Bartonella henselae, (the agent of cat scratch disease),¹⁴ B. tamiae,¹⁰ B. elizabethae, B. rattimassiliensis, and B. tribocorum have been isolated from febrile patients,¹⁵ B. henselae and B. clarridgeiae have been reported in cats,¹¹ and B. clarridgeiae, B. vinsonii subsp. arupensis, B. elizabethae, B. grahamii, B. quintana, B. taylorii, and novel Bartonella genotypes have been found in dogs.^11,16 In rodent species, B. grahamii, B. elizabethae, Candidatus Bartonella thailandensis, B. coopersplainensis, B. phoceensis, B. rattimassiliensis, B. tribocorum, and novel Bartonella genotypes have been detected by culture and polymerase chain reaction (PCR) analysis.^12,13,17 However, little information has been obtained to identify potential arthropod vectors of Bartonella species in Thailand. Bartonella henselae, B. clarridgeiae and B. koehlerae were detected in Ctenocephalides felis fleas removed from cats^18–20 and B. henselae was identified in two C. canis¹⁹ also collected from cats. Furthermore, a Bartonella sp., similar to B. grahamii, was found in a rodent flea, Nosopsyllus fasciatus, obtained from Rattus surifer.¹⁸ Bartonella tamiae DNA has also been found in chigger mites (genera Leptotrombidium, Schoengastia, and Blankarrtia) and in a tick (genus Haemaphysalis) collected from rodents in Thailand, suggesting a potential role for these arthropods in the transmission of B. tamiae.²¹The aim of the current study was to investigate the prevalence of Bartonella species in rodent-associated fleas collected in Khon Kaen Province, Thailand, and to determine what potential role, if any, these fleas may play in the transmission of Bartonella species to individuals residing in this area.For this study, 62 rats (10 R. norvegicus, 9 R. rattus, and 43 R. exulans) were trapped in and around homes in 4 villages, 1 market, and on farm land (a pig farm and 2 rice fields) in Khon Kaen Province, Thailand during May–June 2011 (22 Work involving rodents was conducted as outlined in our approved animal use protocol (#11-003), under the supervision of the Institutional Animal Care and Use Committee of the Division of Vector Borne Diseases.

Table 1

Number of rats trapped per site by species and total number of fleas examined per site, northeastern Thailand

Prevalence of drug-resistant tuberculosis in an HIV endemic area in northern Thailand.

SETTING: Chiang Rai Province in Northern Thailand, where human immunodeficiency virus (HIV) infection has been prevalent since the 1990s. OBJECTIVE: To observe the prevalence of drug-resistant tuberculosis (TB) and investigate the factors related to the level of drug resistance in an HIV endemic area. DESIGN: Population-based surveillance study covering the whole province. METHOD: Drug susceptibility testing was performed at the Thai Ministry of Public Health laboratory for all sputum smear-positive TB patients diagnosed in hospitals in Chiang Rai Province over a 25-month period in 1996-1998. Patient characteristics were obtained through interview by trained personnel. HIV testing was performed with informed consent. RESULTS: Among the 1077 incident patients without previous history of treatment, the proportion of patients with resistance to isoniazid was 13.2%, 10.8% to rifampicin, 15.6% to streptomycin, and 5.8% to ethambutol. Multidrug resistance (MDR), i.e., resistance to at least both isoniazid and rifampicin, was observed in 6.3%. Factors associated with primary MDR-TB were HIV positivity (OR 2.2, 95%CI 1.3-3.9), age <50 years (OR 2.0), and treatment in the provincial hospital (OR 2.3), compared to patients treated in the community and private hospitals. Stratified analysis shows a significantly high prevalence of primary MDR-TB among HIV-positive patients treated in the provincial hospital against HIV-negative patients or HIV-positive patients in other hospitals. CONCLUSION: The prevalence of primary MDR-TB in this area was high. It is necessary to strengthen TB control activities in order to reduce the burden of MDR-TB.     

Prevalence and risk factors of microalbuminuria in patient with diabetic mellitus at northern part referral hospital in Thailand

ObjectiveTo identify prevalence of microalbuminuria and its associated risk factors including old age, hypertension, dyslipidemia, diabetic retinopathy and metabolic syndrome in diabetic patients in Lampang, Thailand.Material and method2231 diabetic patients at Lampang Hospital were recruited in this cross-sectional study. Eye examination, blood pressure and abdominal circumference measurements were done. Blood was collected and checked for glucose, cholesterol, and renal function. Single-spot morning urine was collected for urine albumin–creatinine ratio (UACR). UACR 30–300 mg/g indicated microalbuminuria (by National Kidney Foundation). Multivariate logistic regression method including odds ratio and 95% confidence interval were used to analyze the data.ResultsMicroalbuminuria was found in 27% of the patients. There were clear association between creatinine clearance and microalbuminuria. Older age for every 10 years and diabetic retinopathy had relationship with microalbuminuria in overall and female patients while had no statistically significant in male patients.ConclusionMicroalbuminuria is common in Thai diabetic patients. Older age and history of diabetic retinopathy increase the risk of microalbuminuria in overall and female patients. Screening for albuminuria should be done in every diabetic patient for early prevention of diabetic nephropathy.     

Serological studies of patients with cutaneous and oral-oropharyngeal anthrax from northern Thailand

An outbreak of 52 cases of cutaneous anthrax and 24 cases of oral-oropharyngeal anthrax occurred in rural Northern Thailand in 1982, caused by contaminated water buffalo meat. Microbiologic diagnosis of many of these cases was hindered by delayed presentation for care and by prior antibiotic therapy. In a retrospective investigation, we used enzyme-linked immunosorbent assays to measure antibody titers to components of anthrax edema toxin (edema factor [EF] and protective antigen [PA]), lethal toxin (lethal factor [LF] and PA), and poly-D-glutamic acid capsule. Electrophoretic-immunotransblots (EITB, Western blot) were used to detect antibodies to PA and LF. Nine patients with cutaneous anthrax, 10 patients with oral-oropharyngeal anthrax, and 43 healthy unexposed Thai control villagers were studied. Over all, EITB was positive in 13/18 patients (sensitivity 72%) and 0/43 controls (specificity 100%). The sensitivity of the ELISA was 72% for PA, 42% for LF, 26% for EF, and 95-100% for capsule. Although a few control sera had apparent false positive titers to PA, the specificity of the ELISA confirmed by EITB (100%) demonstrated the applicability of these tests for the diagnosis of anthrax.     

Bartonella species in rodents and shrews in the greater Jakarta area

In February 2004, we captured 221 rodents and shrews in the Greater Jakarta area as part of a study to determine the prevalence of rodent-associated vector-borne infections. Microscopic examination of blood smears revealed 6% (13/218) to be positive for Bartonella spp. The corresponding DNA samples, either from blood blots or frozen spleen pieces and from fleas collected on these animals, were tested for evidence of Bartonella infection by PCR, targeting the portions: 378bp and 930bp of the citrate synthase gene (g/tA). The sequences from our sample clusters with a Peruvian entity, B. phoceensis, B. rattimassiliensis and B. elizabethae, the latter species has been associated with endocarditis and neuroretinitis in humans. As previous analyses have shown, there appears to be little geographic or host consistency with phylogenetic placement. The public health significance of these findings remains to be determined.     

Identification of Bartonella Infections in Febrile Human Patients from Thailand and Their Potential Animal Reservoirs

To determine the role of Bartonella species as causes of acute febrile illness in humans from Thailand, we used a novel strategy of co-cultivation of blood with eukaryotic cells and subsequent phylogenetic analysis of Bartonella-specific DNA products. Bartonella species were identified in 14 blood clots from febrile patients. Sequence analysis showed that more than one-half of the genotypes identified in human patients were similar or identical to homologous sequences identified in rodents from Asia and were closely related to B. elizabethae, B. rattimassiliensis, and B. tribocorum. The remaining genotypes belonged to B. henselae, B. vinsonii, and B. tamiae. Among the positive febrile patients, animal exposure was common: 36% reported owning either dogs or cats and 71% reported rat exposure during the 2 weeks before illness onset. The findings suggest that rodents are likely reservoirs for a substantial portion of cases of human Bartonella infections in Thailand.