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Investigation of Anaplasmosis in Yiyuan County, Shandong Province

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Investigation of Anaplasmosis in Yiyuan County, Shandong Province Asian Pacific Journal of Tropical Medicine (2011)568-572 568 Contents lists available at ScienceDirect Asian Pacific Journal of Tropical Medicine journal homepage:www.elsevier.com/locate/apjtm Document heading doi: Investigation of anaplasmosis in Yiyuan County, Shandong Province, China Lijuan Zhang1* , Feng Cui2, Lingling Wang2, Lingling Zhang2, Jingshan Zhang1, Shiwen Wang1, Shuxia Yang2 1Department of Rickettsiology and Anaplasmosis, National Institute of Communicable Disease Control and Prevention, China CDC, Beijing 102206, China 2Zibo Center for Disease Control & Prevention, Zibo 255026, Shandong, China ARTICLE INFO ABSTRACT Article history: Objective: To investigate the situation of anaplasmosis in Yiyuan county, Shandong Province. Received 11 February 2011 Methods: A total of 26 blood samples from febrile patients suspected of anaplasmosis, 48 blood Received in revised form 11 April 2011 8 10 170 Accepted 15 June 2011 samples from healthy farmers, from dogs, and from goats and ticks were collectedResults: in Available online 20 July 2011 the same area during 2005-2007, and detected by serological and molecular methods. Eight confirmed cases and 6 probable cases were determined using serologic and molecular Anaplasma phagocytophilum A. phagocytophilum Keywords: methods. The seroprevalence of ( ) was 26.7% in healthy cases. Nine out of 10 sheep samples and 7 out of 8 dog samples reacted positively Anaplasma phagocytophilum A. phagocytophilum A. to the antigen. PCR amplification and sequencing of the 16SrRNA of phagocytophilum Field investigation gene showed that some samples from patients, goats and ticks were 100% Rickettsia Rickettsia typhi Orientia tsutsugamushi identical. The seroprevalence of was 22.9%, 6.3%, Rickettsia sibirica Coxiella burnetii Bartonella henselae Borrelia Prevalence 27.1%, 18.8%, 31.3%, and Yiyuan County burgdorferi Conclusions: 41.6%. It is important to make differential diagnosis of febrile patients Shandong Province China and to apply treatment with specific antibiotics. It is needed to enforce essential prevention and control measures including tick control and to improve sanitation conditions. [6-12] 2006 1. Introduction including Korea, Japan and China . In , an unusual nosocomial human-to-human transmission was demonstrated by PCR and seroconversion in Anhui Human granulocytic anaplasmosis (HGA) and human Province, China[13]. As a result of these events, a general monocytic ehrlichiosis (HME) are acute and febrile tick- retrospective laboratory survey of suspected HGA cases borne rickettsial diseases caused by the bacterium collected from 2004-2006 in Jiangsu, Zhejiang, Shandong Anaplasma phagocytophilum A. phagocytophilum ( ) and and Hubei Provinces was undertaken by the Department Ehrlichia chaffeensis E. chaffeensis ( ), respectively[1,2]. As of Rickettsiology of China and local health centers during other rickettsioses, the prevalence of these diseases are 2006-2007. In this report, we outlined the findings from found worldwide, but mostly in the USA and Europe[3-5]. Yiyuan County of Shandong Province, China. Recently, serologic and molecular evidences of these Yiyuan is one of the poorest counties in Shandong diseases had been reported in some Asian countries, Province. It is reported that incidence of unknown endemic febrile (>38 曟) has increased from March to October every [14] year . Typical clinical characteristics of these cases *Corresponding author: Lijuan Zhang, Department of Rickettsiology, National Institute of Communicable Disease Control and Prevention, China CDC, PO Box 5, include high fever, headaches, myalgias, hematological and Changping, Beijing 102206, China. biochemical abnormalities,leucopenia, thrombocytopenia, Tel: +86 23 58900780 Fax: 86 23 58900780 mild/moderately elevated hepatic transaminase levels, E-mail:[email protected] and (in some cases) high rates of urinary protein. Some of Foundation project: This study was supported by the National Basic Research Program of China (973 Program) 2010CB530200(2010CB530206); the National these patients ultimately die due to multi-organ failure. Natural Science Foundation of China (No. 30771854), National Key Science and Laboratory tests of Dengue fever, scrub typhus, Lyme Technology Projects of China (Project No. 2008ZX10004-008). disease and epidemic hemorrhagic fever were performed Lijuan Zhang et al./Asian Pacific Journal of Tropical Medicine (2011)568-572 569 for each patient, but no positive result was obtained. infected healthy controls from the same area. Informed Based on field epidemiology surveys, the occurrence of the’ consent was obtained before data collection. Approval disease was associated with poor sanitation of the patient s OR CI P for the study was obtained from the Shandong’ provincial living room ( : 12.000, 95% : 1.990-72.354, <0.05), institutional review board. Patients information was co-habitationOR withCI sheep and otherP domestic animals obtained, including name, age, gender, occupation, and 6 400 95 : 1 944 21 072 0 05 ( : . OR, % . -CI . , < . ), untreatedP open history of tick bite/exposure. The mean age of the suspected wounds ( : 6.333, 95% : 1.501-26.732, <0.05), wild ’ OR CI cases was 59 years old, and all were farmers, including 10 5 909 95 1 690 grass aroundP the patient s house ( : . , % : . - male (mean age 60 years old) and 16 female (mean age 59 20 660 0 05 . , < . ),OR and working onCI a field for moreP than one years old). They all had high fever (>38 ), leucopenia, [11] 曟 hour per day ( : 5.250, 95% : 1.407-19.593, <0.05) . thrombocytopenia, and mild/moderately elevated hepatic Local epidemiologists have suggested that the disease is a transaminase levels; 65% had headache and myalgias, 20% type of tick-related natural foci disease, because ticks are had localized enlarged lymph nodes with lymphangitis and the most abundant disease vectors in domestic and wild digestive system involvement, 5% had eye socket ache, animals in these areas. Hence, we first evaluated emerging and 5% presented a rash. Two patients died of multi-organ tick-borne rickettsial diseases based on clinical features, A. phagocytophilum E. chaffeensis failure. The mean age of 48 was 50 years old, including 26 particularly with and . male (mean age = 52 years old) and 22 female (mean age =48 26 In this survey, we retrospectively investigated blood years old), and 82% were farmers. 2004 2006 samples from suspected cases collected from - . Blood (5 mL) was collected from each suspected patient at We also evaluated blood samples from 48 healthy controls the acute stage of illness; a second samples were collected and predominant domestic animals (including 10 goats and from 14 patients at convalescent phase of illness (3 or 4 8 dogs) using serologic methods. In addition, tick samples weeks after onset of illness). Separated serum was used for from the same areas where the suspected patients lived were indirect immunofluorescence assays and DNA was extracted identified by molecular methods. from red blood cells for PCR amplification. Blood (5 mL) from controls was collected to detect IgG-specific antibody 2. Materials and methods A. phagocytophilum E. chaffeensis titers of and℃ . All samples were temporally stored at -20 . 2.1. Study area and populations 2.3. Diagnosis standard The study was conducted in 633 administrative hamlets, 33 administrative villages of 11 towns in Yiyuan County, China. A positive diagnosis was made if any one of the following These were all located in the mountainous areas of Yimeng conditions was met: 1) typical epidemiologic history of tick ° ’ ° ’′ ° ’ ° ’’ 2 (117 54 -118 31 E and 35 55 -36 23 N) and had a total exposure/bite, or outdoor activity; ) clinical manifestation℃ population of 555 000, among which farmers accounted for of tick-borne rickettsioses, such as high fever (>38 ), 85%. Foothills with heavy wild grass, low, middle, and high headache, myalgias,localized enlarged lymph nodes, or lymphangitis; 3) laboratory-based diagnosis of leucopenia, upland were the most prevalent geographic features. The℃ area is seasonal, with an average temperature of 11.9 , thrombocytopenia, or mild/moderately elevated hepatic and average annual rainfall of 720.8 mm. Besides planting transaminase levels. crops, most common human occupations are livestock- 2.4. Collection of ticks from goats and dogs raising activities (predominantly by women) and active outdoor activities such as fishing and fruit tree farming (predominantly by men). Since many younger had moved to One hundred and fifty ticks from goats and 20 ticks from ne＞arby urban areas, the major in families are older people dogs were obtained in different farmer families from different ( 50 years old). Goats (Boer sheep imported from America, villages. A total of 34 groups of ticks (5 ticks from the same and native black goats) farming plays an important role in animal as a group) were homogenized and their DNA was each family, and pen-breeding in courtyards is the most PCR Haemaphysalis longicornis H. longicornis extracted for amplification. Local slaughterhouses were common method. ( ) 10 Haemaphysalis concinna H. concinna selected for collecting animal blood, where goat blood and ( ) ticks are samples and 8 dog blood samples were obtained for serologic predominant species in these areas[15] and actively grow from and molecular analyses. April to October. Higher density of these ticks are found in 2.5. Serologic testing grass on hill areas. Ticks are common external parasites of goats and dogs from these areas. All serum samples
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