Historical geographic overlap of human rickettsiosis with animal reservoirs in Juiz de Fora, Minas Gerais, Brazil / Sobreposição geográfica histórica da riquetsiose humana com reservatórios animais em Juiz de Fora, Minas Gerais, Brasil

Background and objectives: we investigated the existence of a historical geographic overlap between the location of spotted fever group rickettsiosis human cases, a disease caused by the gram-negative bacterium Rickettsia sp., and that of mammalian reservoirs, specifically domestic horses and capybaras, in the urban perimeter of the city of Juiz de Fora, Minas Gerais, Brazil. Methods: cases of human rickettsiosis that occurred during a period of 17 years (2003-2020) were geolocated and the distribution of cases in time and geographic space was assessed using 1st and 2nd order geospatial association indicators. We also analyzed the overlap between the locations of human rickettsiosis cases and the area of occurrence of domestic horses and capybaras. Results: men were diagnosed more often than women, but a large proportion of affected women died. The results indicate an aggregation of human rickettsiosis cases in time (cases tend to occur close to each other at each epidemic event) and in geographic space (cases are concentrated in a specific geographic region of the urban perimeter). Human cases seem to be more associated with city regions with: i) higher local frequency of domestic horses and not capybaras; ii) lower rates of family development. Conclusion: it is suggested that, in the local epidemiological scenario, domestic horses appear to be the main sources of the rickettsia infecting humans, not capybaras.(AU)

Justificativa e objetivos: foi investigada a existência de uma sobreposição geográfica histórica entre a localização dos casos de riquetsiose humana do grupo da febre maculosa, um grupo de doenças causadas pela bactéria Gram-negativa Rickettsia sp., e dos reservatórios mamíferos, especificamente cavalos domésticos e capivaras, no perímetro urbano do município de Juiz de Fora, Minas Gerais, Brasil. Métodos: foram geolocalizados os casos de riquetsiose humana ocorridos durante um período de 17 anos (2003-2020), sendo avaliada a distribuição dos casos no tempo e espaço geográfico através de indicadores de associação geoespacial de 1a e 2a ordem. Também analisamos a superposição dos locais dos casos de riquetsiose humana com a área de ocorrência de cavalos domésticos e capivaras. Resultados: homens foram diagnosticados mais frequentemente que as mulheres, mas grande proporção das mulheres acometidas faleceu. Os resultados indicam uma agregação dos casos de riquetsiose humana no tempo (os casos tendem a ocorrer próximos entre si a cada evento epidêmico) e no espaço geográfico (os casos se concentram em uma região geográfica específica do perímetro urbano). Os casos humanos aparentam ser mais associados às regiões da cidade com: i) maior frequência local de cavalos domésticos e não das capivaras; ii) menores índices de desenvolvimento familiar. Conclusão: sugere-se que, no cenário epidemiológico local, são os cavalos domésticos que aparentam ser as principais fontes da riquétsia infectando os humanos, não as capivaras.(AU)

Justificación y objetivos: investigamos la existencia de una superposición geográfica histórica entre la localización de casos de rickettsiosis exantemáticas humana, enfermedad causada por la bacteria gramnegativa Rickettsia sp., y la de mamíferos reservorios, específicamente caballos domésticos y capibaras, en el perímetro urbano de la ciudad de Juiz de Fora, Minas Gerais, Brasil. Métodos: se geolocalizaron los casos de rickettsiosis humana ocurridos durante un período de 17 años (2003-2020), y se evaluó la distribución de casos en el tiempo y espacio geográfico utilizando indicadores de asociación geoespacial de 1er y 2do orden. También analizamos la superposición entre las ubicaciones de los casos de rickettsiosis humana y el área de ocurrencia de los caballos domésticos y capibaras. Resultados: los hombres fueron diagnosticados con más frecuencia que las mujeres, pero una gran proporción de mujeres afectadas fallecieron. Los resultados indican una agregación de casos de rickettsiosis humana en el tiempo (los casos tienden a ocurrir cerca uno del otro en cada evento epidémico) y en el espacio geográfico (los casos se concentran en una región geográfica específica del perímetro urbano). Los casos humanos parecen estar más asociados con regiones urbanas con: i) mayor frecuencia local de caballos domésticos y no de capibaras; ii) menores tasas de desarrollo familiar. Conclusión: se sugiere que, en el escenario epidemiológico local, los caballos domésticos parecen ser las principales fuentes de la rickettsia que infecta a los humanos, no los capibaras.(AU)

Investigation on the infection of spotted fever group Rickettsiae and Rickettsia mooseri of wild rodents in the field of plague foci in Western Yunnan / 中华地方病学杂志

Objective:To investigate the infection of spotted fever group Rickettsiae (SFGR) and Rickettsia mooseri ( R.mooseri) of wild rodents in the field of plague foci in Western Yunnan. Methods:The DNA of liver samples of 2 512 wild rodents captured from the plague foci in Lianghe County, Jianchuan County and Yulong County in Western Yunnan from 2015 to 2016 was extracted by magnetic bead method, and the heat shock protein groEL gene primers were used for nested PCR amplification. Gene sequence splicing and Blast homology comparison were performed using DNAStar 7.1 software and GenBank of the National Center for Biotechnology Information (NCBI) of the United States, respectively, and DNAStar 7.1 and MEGA 6.0 softwares were used to construct phylogenetic trees.Results:The wild rodents infected with SFGR were Mus pahari, Rattus steini, Crocidura attenuata and Suncus murinus (one for each), with a total infection rate of 0.16% (4/2 512); no R.mooseri infection was detected. The SFGR infection rates of wild rodents in the plague foci of Lianghe County and Jianchuan County were 0.49% (3/611) and 0.10% (1/1 029), respectively; no SFGR infection was detected in the wild rodents in the plague foci of Yulong County. The homology analysis showed that the homology between SFGR positive samples and reference sequences was 95.45%-100.00%; some of the groEL gene sequences were highly similar among the four positive samples, and the homology was 89.60%-97.40%. Sequence evolution analysis showed that the sequences of three SFGR positive samples from the plague focus in Lianghe County were clustered in the same branch, and the homology reached 94.40%-97.40%; one positive sample sequence from the plague focus in Jianchuan County was clustered in one branch. Conclusion:SFGR infection rate of wild rodents in the field of plague foci in Western Yunnan is low, and no R.mooseri infection is found.

A new record of Rickettsia japonica in ticks infesting a Burmese ferret-badger in Thailand

@#Ticks are important vectors of arthropod-borne diseases and they can transmit a wide variety of zoonotic pathogens to humans, domestic and wild animals. Rickettsia japonica is a member of SFG rickettsiae causing Japanese spotted fever (JSF) and can transmit to humans via infected ticks. In this study, we report the first case of Rickettsia japonica in Haemaphysalis hystricis tick collected from a roadkill Burmese ferret-badger (Melogale personata) in Loei province, northeastern Thailand. According to the DNA sequences and phylogenetic analyses of the outer membrane protein A and B genes (ompA and ompB), the detected R. japonica was identical to those found in JSF patients in Korea, Japan, and China, and closely related to Rickettsia detected by ompA in a tick from Thailand. Further study on the prevalence of R. japonica and diversity of mammalian reservoir hosts will be useful to gain a better understanding of JSF epidemiology.

Investigation and research on ticks carrying spotted fever group rickettsia in the border area of Tumen River Basin / 中华预防医学杂志

Objective@#To understand the situation and genotype distribution of spotted fever group rickettsia (SFGR) in the border area of Tumen River Basin in free ticks in Yanbian Korean Autonomous Prefecture (Yanbian Prefecture), Jilin Province. @*Methods@#From April to September, 2017, ticks were collected using flagging method from Hunchun, Tumen, Helong and Longjing cities in the Tumen River basin of Yanbian Prefecture. Outer membrane protein A (ompA) was detected by Polymerase Chain Reaction (PCR), then, the species were identified by gene sequencing and analyzed systematically. The positive rate of pools and MIR(minimum infection rate per 100 ticks,MIR) of SFGR were calculated, and the difference of positive rate of pools among ticks with different characteristics was compared by Chi-square test. @*Results@#A total of 3 079 ticks were collected and divided into 536 pools. The positive rate of pools of SFGR nucleic acid was 39.7% (213 pools). The MIR of SFGR was 6.9%.The positive rate of pools of SFGR in Dermacentor silvarum, Haemaphysalis concinna, Haemaphysalis japonica, Haemaphysalis longicornis and Ixodes persulcatus were 80.4% (41/51), 14.0% (25/179), 20.2% (18/89), 78.9% (101/128) and 25.9% (21/81), and the difference was statistically significant (P<0.001). There was statistical difference in the positive rate of pools of SFGR in developmental stages of ticks (P<0.001); the positive rate of pools of female adults, male adults, nymph and larvae were 36.4% (95/261), 34.2% (67/196), 56.3% (40/71) and 7/8, and the MIR was 7.9%, 7.7%, 4.9% and 3.5%. The five genotype was detected which was Candidatus Rickettsia longicornii, Rickettsia raoultii, Rickettsia heilongjiangensis, Candidatus Rickettsia tarasevichiae,Rickettsia monacensis and have 98%-100% homology with known gene sequences. Candidatus Rickettsia longicornii, Rickettsia raoultii, Rickettsia heilongjiangensis and Candidatus Rickettsia tarasevichiae showed close evolutionary relationship with known specie (have 98%-100% homology with known gene sequences); Rickettsia monacensis showed Far from evolutionary relationship with known species (have 98% homology with known gene sequences). @*Conclusion@#SFGR infection of ticks is common in the border areas of the Tumen River Basin. There was high diversity in SFGR species and tick species in the areas surveyed.

Fiebre manchada de evolución fatal en la provincia de Salta / Fatal spotted fever in Salta province

Describimos el caso de un varón de 17 años oriundo de la ciudad de Salta quien, 10 días después de visitar una zona rural de la provincia homónima, ingresó a nuestro hospital por convulsiones febriles. Durante la internación presentó exantema seguido de disfunción orgánica múltiple, la que evolucionó rápidamente hacia shock séptico irreversible y muerte a las 48 horas de su admisión. El diagnóstico serológico -altos títulos de IgM e IgG anti-Rickettsia spp. por inmunofluorescencia indirecta- arribó post mortem. Las rickettsiosis del grupo de las fiebres manchadas son transmitidas por garrapatas, tienen distribución global y en varios países continúan siendo subdiagnosticadas debido a una baja sospecha clínica. En las provincias del noroeste argentino se agrega la carencia de un laboratorio regional capacitado para efectuar el diagnóstico etiológico. Esta limitación es crítica porque en esa región del país ya ha sido documentada la presencia de las formas graves de la enfermedad, usualmente debidas a R. rickettsii. Dado que las fiebres manchadas se presentan como sindromes febriles inespecíficos y los componentes del ciclo enzoótico están presentes en vastas áreas geográficas, incluso en algunas aún no consideradas endémicas para rickettsiosis, su diagnóstico nunca debe ser subestimado. Con el tratamiento antibiótico adecuado instaurado en tiempo oportuno, el pronóstico de este grupo de enfermedades potencialmente mortales mejora en forma drástica.

We describe the case of a 17-year-old male patient living in Salta city who, 10 days after visiting a rural area in Salta province, was hospitalized for febrile seizures. Shortly after admission, he developed an exanthema followed by a multiple organ dysfunction that evolved to irreversible septic shock followed by death 48 hours after admission. Serological diagnosis -high IgM and IgG anti-Rickettsia spp. antibody titres as detected by indirect immunofluorescence- arrived post mortem. Spotted fever group rickettsioses are tick-borne diseases distributed worldwide and continue being under diagnosed in several countries mainly due to a low clinical suspicion. In the north-western provinces of Argentina there is also the limitation of not counting with a regional laboratory to perform the etiological diagnosis. This is crucial because the severe forms of the disease, which are commonly caused by R. rickettsii, have been already documented in the region. Given that spotted fevers have broadly unspecific febrile presentations and the components of the enzootic cycle are present even in geographic areas not yet considered to be endemic for tick borne diseases, their diagnosis should not be underestimated. If the adequate antibiotic treatment is administered timely, the prognosis of this group of life-threatening diseases improves drastically.

Seroprevalence of Rickettsia rickettsii and Rickettsia parkeri in dogs during a Brazilian Spotted Fever outbreak in the State of Rio de Janeiro / Soroprevalência de Rickettsia rickettsii e Rickettsia parkeri em cães durante um surto de Febre Maculosa Brasileira no Estado do Rio de Janeiro

The present paper is the first to perform this evaluation in dogs from the cities of Natividade, Porciuncula and Varre-Sai. The aim of this study is to search for Spotted Fever Group Rickettsia in canine sera using indirect immunofluorescence assay and to identify the probable causative agent of sera reactions in animals. Of the 253 sampled canines, 67.59% (171/253) were seroreactive for Rickettsia rickettsii and 11.07% (28/253) for Rickettsia parkeri, both in dilution 1:64. Titration of tested sera against R. rickettsii antigens reached 1:131.072 and, for R. parkeri, 1:4.096. We conclude that dogs are important sentinels for R. rickettsii infection, and can be infected regardless of sex, age, the habit of visiting woodlands or being in direct contact with equines and capybaras. Serological diagnosis has highlighted many dogs infected by R. rickettsii, and ambient conditions, such as the presence of flowing water bodies, was important for the occurrence of Brazilian Spotted Fever in the northwestern of Rio de Janeiro State.(AU)

O presente trabalho é o primeiro a ser realizado com cães nos municípios de Natividade, Porciúncula e Varre-Sai e tem por objetivo pesquisar Rickettsias do Grupo da Febre Maculosa em soros de cães, por meio da reação de imunofluorescência indireta, e identificar o provável agente causador da reação sorológica nos animais. Dos caninos amostrados, 67,59% (171/253) foram sororreativos para Rickettsia rickettsii e 11,07% (28/253) para Rickettsia parkeri, ambos em diluição de 1:64. A titulação dos soros testados contra antígenos de R. rickettsii chegou a 1:131.072, e para R. parkeri, 1:4.096. Conclui-se que cães são importantes sentinelas para a infecção por R. rickettsii, independente de sexo, idade, hábito de visitar ambientes florestais ou de estarem em contato direto com equinos e capivaras. O diagnóstico sorológico permitiu evidenciar muitos cães infectados por R. rickettsii, e condições ambientais, como a presença de áreas ribeirinhas, foram importantes para a ocorrência de Febre Maculosa Brasileira na região noroeste do Estado do Rio de Janeiro.(AU)

Multispacer Typing (MST) of Spotted Fever Group Rickettsiae Isolated from Humans and Rats in Chengmai County, Hainan Province, China

Spotted fever caused by spotted fever group rickettsiae (SFGR) is found throughout China. During 2007–2008, 28 human SFGR isolates and 34 rat SFGR isolates including 15 isolates from <i>Rattus fulvescens</i>, 5 isolates from <i>R. edwardsi</i>, 7 isolates from <i>Callosciurus erythraeus roberti</i> and 7 isolates from <i>Dremomys rufigenis</i>) were obtained from L929 cell culture. Previous research indicated that the 62 strains of SFGR mentioned above shared not only the same serophenotype but also 100% of identity sequences of 16S rRNA, <i>gltA</i>, <i>ompA</i>, <i>groEL</i> and 17KD, which enabled us to apply multispacer typing (MST) to the 62 SFGR isolates in the study. Six primer pairs, which were used for typing of <i>Rickettsia rickettsii</i> and <i>Rickettsia conorii</i>, were chosen, and the results exhibited greater nucleotide polymorphisms among the 62 isolates tested. A total of 48 distinct genotypes were identified. The dominant genotype, represented by h3 isolates, accounted for 21.7% (13/60) of the isolates tested, and the remaining 47 genotypes were all unique. Phylogenetic analysis showed that all the 48 genotypes could be classified in the same clade, while the genetically related strain, <i>R. heilongjiangensis</i>, was close but not the same as the cluster. We concluded that the genetically diverse of spotted fever group rickettsiae strains are endemic in Chengmai County, Hainan Province, China.

Molecular detection of spotted fever group rickettsia in Dermacentor silvarum from the Xunke Area,China / 中国人兽共患病学报

To identify the prevalence and the distribution of ticks infected with spotted fever group rickettsiae (SFGR) in Xunke Area of Heilongjiang Province ,China ,partial outer membrane protein A gene (ompA) and citrate synthase gene (gltA) specific fragments were assessed using the PCR method .The positive products were sequenced .Result showed that the pres-ence of SFGR was 14 of 60 in detection Dermacentor silvarum cases ,while the overall positive rate was 23 .33% .Its nucleotide sequence of ompA showed 99 .3% and 99 .18% similarity with nucleotide sequence of Rickettsia sp .JL-02 and Rickettsia rao-ultii respectively .And the evolutionary positions of SFGR species were similar with Rickettsiamontana and Rickettsiamassili-ae .It's concluded that the nature focus of tick-borne spotted fever did exist in the area of Xunke Area of Heilongjiang Province , China .

Multispacer typing (MST) of spotted fever group rickettsiae isolated from humans and rats in Chengmai County, Hainan Province, China

Spotted fever caused by spotted fever group rickettsiae (SFGR) is found throughout China. During 2007—2008, 28 human SFGR isolates and 34 rat SFGR isolates including 15 isolates from <i>Rattus fulvescens</i>, 5 isolates from <i>R.edwardsi</i>, 7 isolates from <i>Callosciurus erythraeus roberti</i> and 7 isolates from <i>Dremomys rufigenis</i>) were obtained from L929 cell culture. Previous research indicated that the 62 strains of SFGR mentioned above shared not only the same serophenotype but also 100% of identity sequences of 16S rRNA, <i>gltA</i>, <i>ompA</i>, <i>groEL</i> and 17KD, which enabled us to apply multispacer typing (MST) to the 62 SFGR isolates in the study. Six primer pairs, which were used for typing of <i>Rickettsia rickettsii</i> and <i>Rickettsia conorii</i>, were chosen, and the results exhibited greater nucleotide polymorphisms among the 62 isolates tested. A total of 48 distinct genotypes were identified. The dominant genotype, represented by h3 isolates, accounted for 21.7% (13/60) of the isolates tested, and the remaining 47 genotypes were all unique. Phylogenetic analysis showed that all the 48 genotypes could be classified in the same clade, while the genetically related strain, <i>R.heilongjiangensis</i>, was close but not the same as the cluster. We concluded that the genetically diverse of spotted fever group rickettsiae strains are endemic in Chengmai County, Hainan Province, China.

Rickettsia parkeri: a Rickettsial pathogen transmitted by ticks in endemic areas for spotted fever rickettsiosis in southern Uruguay / Rickettsia parkeri: patógeno rickettsial transmitido por garrapatas en áreas endémicas de rickettsiosis por fiebre manchada en el sur de Uruguay

At first Rickettsia conorii was implicated as the causative agent of spotted fever in Uruguay diagnosed by serological assays. Later Rickettsia parkeri was detected in human-biting Amblyomma triste ticks using molecular tests. The natural vector of R. conorii, Rhipicephalus sanguineus, has not been studied for the presence of rickettsial organisms in Uruguay. To address this question, 180 R. sanguineus from dogs and 245 A. triste from vegetation (flagging) collected in three endemic localities were screened for spotted fever group (SFG) rickettsiosis in southern Uruguay. Tick extracted DNA pools were subjected to PCR using primers which amplify a fragment of the rickettsial gltA gene. Positive tick DNA pools with these primers were subjected to a second PCR round with primers targeting a fragment of the ompA gene, which is only present in SFG rickettsiae. No rickettsial DNA was detected in R. sanguineus. However, DNA pools of A. triste were found to be positive for a rickettsial organism in two of the three localities, with prevalences of 11.8% to 37.5% positive pools. DNA sequences generated from these PCR-positive ticks corresponded to R. parkeri. These findings, joint with the aggressiveness shown by A. triste towards humans, support previous data on the involvement of A. triste as vector of human infections caused by R. parkeri in Uruguay.

Inicialmente, Rickettsia conorii fue señalada como el agente causal de la fiebre manchada en Uruguay, diagnosticada mediante pruebas serológicas. Posteriormente, Rickettsia parkeri fue detectada mediante técnicas moleculares en garrapatas Amblyomma triste colectadas sobre humanos. El vector natural de R. conorii, Rhipicephalus sanguineus, no ha sido estudiado en cuanto a rickettsias en Uruguay. Para abordar este tema, 180 R. sanguineus fueron colectados sobre perros y 245 A. triste sobre vegetación en tres localidades consideradas endémicas para fiebres manchadas en el sur de Uruguay. El ADN de las garrapatas fue extraído en pools y sometido a una primera PCR utilizando cebadores que amplifican un fragmento del gen gltA, presente en prácticamente todas las especies de Rickettsia. Las muestras positivas fueron sometidas a una segunda PCR con cebadores que amplifican un fragmento del gen ompA, presente sólo en rickettsias del grupo de las fiebres manchadas (GFM). No se detectó ADN rickettsial en R. sanguineus. Sin embargo, muestras de A. triste fueron positivas a rickettsiales en dos de las tres localidades estudiadas, con prevalencias de pools positivos del 11.8 y 37.5% respectivamente. La secuenciación del ADN evidenció la presencia de R. parkeri. Basados en estos resultados junto a los anteriores y la agresividad de A. triste hacia los humanos, se concluye que esta garrapata es vector de rickettsiosis humana por R. parkeri en Uruguay.

Dynamic investigation on the co-infection status of two pathogens in ticks from tourist point in Heilongjiang province / 中华流行病学杂志

Objective To monitor the co-infection status of Borrelia burgdorferi sensu lato (R.b.s.1) and spotted fever group Rickettsia (SFGR) in tourist areas of Heilongjiang province.Methods Polymerase chain reaction (PCR) was used to detect the 5S-23S rRNA intergenic spacer of B.b.s.1 and ompA of SFGR in ticks,dynamically collected from tourist areas of Heilongjiang province in 2010.Amplification products from positive ticks were sequenced,and phylogenetic analysis was conducted by Mega 5.0 software package.Results 849 ticks were collected from two tourist points,with the dominant ticks in Tiger Mountain and Jingpo Lake were Ixodes persulcatus and Haemaphysalis concinna.Regarding the Ixodes persulcatus from Tiger Mountain,the infection rates of B.b.s.1 and SFGR were 26.15％ and 10.05％.The infection rate of SFGR was 13.33％ in Haemaphysalis concinna and the B.b.s.1 was tndiscovered in the same ticks from Jingpo Lake.However,the co-infection could only be detected in Ixodes persulcatus of both tourist areas.Surveillance data showed that the major ticks were more likely to be appeared in July at Tiger Mountain and in June at Jingpo Lake.Data from the sequence analysis on B.b.s.1 showed that the B.b.s.1 in tourist areas could be classified into three different genotypes,other than B.garinii and B.afzelii.We first detected B.valaisiana-like group genotype in northeast of China.Results from the sequence analysis of SFGR positive products showed that the two DNA sequences of newly detected agents were completely the same as Rickettsia sp.HL-93 which was detected in Hulin and Rickettsia sp.H820 found in northeast,China.Conclusion The co-infection of B.b.s.1 and SFGR was detected in ticks from the tourist areas of Heilongjiang province,and data from the sequencing of specific fragment showed that various kinds of genotypes existed in this area.However; the rates of co-infectionitis-different according to environment,time and population that contributed to the kinds of and the index of ticks existed in the surveys points,also the infection rate of the ticks was studied.

Characterization of rickettsia rickettsii in a case of Fatal Brazilian spotted fever in the city of Rio de Janeiro, Brazil

A lethal case of Brazilian spotted fever (BSF) is presented. Clinical features were initially of gastrointestinal involvement and evolved with progression to septic shock, meningoencephalitis and death on the 6th day of illness. Indirect immunofluorescence assay (IFA) for spotted fever group rickettsia (SFGR) was non-reactive. Diagnosis was confirmed by the polymerase chain reaction (PCR) and the nucleotide sequencing of a fragment of the ompA gene showed 100 percent homology to Rickettsia rickettsii. BSF has not been reported in the city of Rio de Janeiro in the last three decades, and the present description should alert the clinicians to its presence in urban Rio de Janeiro, and to the differential diagnosis with dengue fever, gastroenteritis, leptospirosis and bacterial septic shock, among others.

Detection of co-infection with Lyme spirochetes and Spotted fever group rickettsiae in a group of Haemaphysalis longicornis / 中华流行病学杂志

Objective The present study was conducted to investigate the infection of Lyme disease, Spotted fever, Ehrlichiosis (anaplasmosisin) in wild animals and ticks in the mountain areas of Zhejiang province. Methods Nested polymerase chain reaction was used to amplify specific DNA sequences of Lyme spirochetes, Spotted fever group rickettsiae, Ehrlichia (anaplasma) from samples of mice and ticks. Results 14 positive samples were identified from 121 mice and 105 groups of ticks. Among mice samples, one positive 5S-23S rDNA intergenic spacer of Borreia burgdorferi and two 5' fragments of Ehrlichia (anaplasma) 16S rDNA were obtained. 11 positive results were detected from tick samples including three 5S-23S rDNA intergenic spacer regions of Borreia burgdorferi and eight 5' fragments of Spotted fever group rickettsiae outer member protein A gene. One group of adult ticks, Haemaphysalis longicornis, which had been collected from eastern mountain area were detected to have co-infected with Lyme spirochetes and Spotted fever group rickettsiae. The positive sequences of 5S-23S rDNA intergenic spacer and ompA gene were tested and analyzed as Lyme spirochetes while rickettsia which was closely related to Borrelia valaisiana and R. massilliae. Conclusion This was the first report about co-infection of Lyme spirochetes and Spotted fever group rickettsiae found in the same group of adult Haemaphysalis longicornis. It is very important to strengthen the surveillance program on tick-borne infectious disease and their pathogenic in vectors, wild animals and targeted high risk groups and to differentiate the clinical manifestation and diagnosis to extend the knowledge of tick-borne infectious diseases in Zhejiang.