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Sample records for anaplasma phagocytophilum exploits

  1. Molecular Evidence for Anaplasma phagocytophilum in Israel

    PubMed Central

    Massung, Robert F.; Inbar, Moshe; Wallach, Arian D.; Shanas, Uri; Mumcuoglu, Kosta Y.; Waner, Trevor

    2007-01-01

    Sequences from the Anaplasma phagocytophilum 16S rRNA gene were detected in 5 ticks representing 3 species (Hyalomma marginatum, Rhipicephalus turanicus, and Boophilus kohlsi) collected from roe deer (Capreolus capreolus) in Mount Carmel, Israel. The sequences were all identical to those of Ap-variant 1 strain. PMID:18252125

  2. Two Anaplasma phagocytophilum Strains in Ixodes scapularis Ticks, Canada

    PubMed Central

    Krakowetz, Chantel N.; Dibernardo, Antonia; Lindsay, L. Robbin

    2014-01-01

    We developed PCR-based assays to distinguish a human pathogenic strain of Anaplasma phagocytophilum, Ap-ha, from Ap-variant 1, a strain not associated with human infection. The assays were validated on A. phagocytophilum-infected blacklegged ticks (Ixodes scapularis) collected in Canada. The relative prevalence of these 2 strains in I. scapularis ticks differed among geographic regions. PMID:25417645

  3. Anaplasma phagocytophilum from Rodents and Sheep, China

    PubMed Central

    Zhan, Lin; Jiang, Jia-Fu; Zhang, Xiao-Ai; Liu, Yun-Xi; Wu, Xiao-Ming; Zhang, Wen-Yi; Zhang, Pan-He; Bian, Chang-Ling; Dumler, J. Stephen; Yang, Hong; Zuo, Shu-Qing; Chu, Chen-Yi; Liu, Wei; Richardus, Jan H.; Habbema, J. Dik F.

    2010-01-01

    To characterize the strains of Anaplasma phagocytophilum in wild and domestic animals in China, we isolated the organism from rodents and sheep in northeastern China. We isolated 3 strains (2 from rodents and 1 from sick sheep) through propagation in BALB/c mice and then cell culture in HL60 cells. The 3 isolates were identified by Wright-Giemsa staining, immunofluorescence, and electronic microscopy and were characterized by sequence analyses of the 16S rRNA gene, partial citrate synthase gene, major surface protein 4 gene, and heat shock protein gene. The multiple sequences of the 3 isolates were identical to each other but different from all known strains from other countries. The public health and veterinary relevance of the isolates deserves further investigation. PMID:20409364

  4. Anaplasma phagocytophilum seroprevalence in equids: a survey in Sicily (Italy).

    PubMed

    Giudice, Elisabetta; Giannetto, Claudia; Furco, Vincenzo; Alongi, Angela; Torina, Alessandra

    2012-08-01

    This study was undertaken to determine the prevalence of Anaplasma phagocytophilum infection in Equidae and investigate the possibility of exposure to the organism in Sicily (Southern Italy). During the study blood samples were collected in horses and donkeys housed in five of the nine provinces of Sicilian Island. Of 133 horses and 100 donkeys tested, respectively 9.0% and 6.0% were seroactive (IFAT) with A. phagocytophilum antigen. In only 4.7% of the horses, specific A. phagocytophilum DNA was recorded; in donkey, Anaplasma DNA was not found. Our results indicate a low prevalence of A. phagocytophilum in Sicilian equids. This condition does not justify the exclusion of equids from prophylactic plans for this multihost pathogen infection, a zoonosis with a wide distribution in other European countries. However, further studies are necessary to elucidate the possible mechanisms that involve the Equidae as host of this pathogen.

  5. Wild animals as reservoirs of Anaplasma phagocytophilum for humans

    PubMed

    Dzięgiel, Beata; Adaszek, Łukasz; Winiarczyk, Stanisław

    Anaplasma phagocytophilum is a tick-transmitted obligate-intracellular gram-negative bacteria that causes emerging human zoonosis. A. phagocytophilum is transmitted by Ixodid ticks. Recent studies suggest that wild animals may be reservoirs of A. phagocytophilum for humans. The organism infects and survives within neutrophils. The infection diagnosis is based on the detection of morulae within granulocytes of peripheral blood, results of serological tests and detection of the DNA of A. phagocytophilum using specific polymerase chain reaction assays (PCR). A. phagocytophilum in most cases is transmitted to people by tick bites, but sometimes direct contact with infected blood may cause human granulocytic anaplasmosis (HGA). The possibility of infection should be taken into consideration at each occurrence of heavy disease symptoms after people come into contact with ticks.

  6. Anaplasma phagocytophilum in ruminants in Europe.

    PubMed

    Woldehiwet, Zerai

    2006-10-01

    The agent that causes tick-borne fever (TBF) in sheep was first described in 1940, 8 years after the disease was first recognized in Scotland. The same agent was soon shown to cause TBF in sheep and pasture fever in cattle in other parts of the UK, Scandinavia, and other parts of Europe. After the initial use of the name Rickettsia phagocytophila, the organism was given the name Cytoecetes phagocytophila to reflect its association with granulocytes and its morphological similarity with Cytoecetes microti. This name continued to be used by workers in the UK until the recent reclassification of the granulocytic ehrlichiae affecting ruminants, horses, and humans as variants of the same species, Anaplasma phagocytophilum. TBF and pasture fever are characterized by high fever, recurrent bacteremia, neutropenia, lymphocytopenia, thrombocytopenia, and general immunosuppression, resulting in more severe secondary infections such as tick pyemia, pneumonic pasteurellosis, listeriosis, and enterotoxemia. During the peak period of bacteremia as many as 90% of granulocytes may be infected. The agent is transmitted transtadially by the hard tick Ixodes ricinus, and possibly other ticks. After patent bacteremia, sheep, goats, and cattle become persistently infected "carriers," perhaps playing an important role in the maintenance of infection, in the flock/herd. Little is known about how efficiently ticks acquire and maintain infection in ruminant populations or whether "carrier" domestic ruminants play an important role as reservoirs of infection, but deer, other free-living ruminants, and wild rodents are also potential sources of infection. During the late 1990s serological evidence of infection of humans was demonstrated in several European countries, creating a renewed interest and increased awareness of the zoonotic potential of TBF variants. More recently, a few cases of human granulocytic anaplasmosis (HGA) have been reported in some European countries, but it remains to

  7. Anaplasma phagocytophilum in questing Ixodes ricinus ticks in southwestern Finland.

    PubMed

    Sormunen, Jani J; Penttinen, Ritva; Klemola, Tero; Vesterinen, Eero J; Hänninen, Jari

    2016-12-01

    Anaplasma phagocytophilum is the causative agent of an emerging tick-borne disease, human granulocytic anaplasmosis. While the bacterium has been reported from questing ticks in neighboring Sweden, Norway and Russia, the few surveys regarding questing ticks in Finland have thus far been negative. In the current study, the prevalence of A. phagocytophilum in Ixodes ricinus populations was evaluated in several study localities around southwestern Finland during 2013-2014. Some of these populations were previously screened and found negative for A. phagocytophilum in 2000. A total of 3158 I. ricinus collected by blanket dragging were screened for Anaplasma spp. using qPCR. Anaplasma were detected in 9.2% of adult ticks (n = 87) and 3.1% of nymphs (n = 979). All larval samples were negative for infection. All Anaplasma-positive samples were identified as A. phagocytophilum by sequencing. This is, to the best of our knowledge, the first report of the pathogen from questing ticks in Finland. Furthermore, the pathogen was detected from several localities found negative during the previous screening 13 years earlier.

  8. Prevalence of Anaplasma phagocytophilum in North Carolina Eastern Black Bears ( Ursus americanus ).

    PubMed

    Westmoreland, Lori S H; Stoskopf, Michael K; Maggi, Ricardo G

    2016-10-01

    We detected Anaplasma phagocytophilum by DNA amplification in whole blood from free-ranging, hunter-killed American black bears ( Ursus americanus ) from the east coast of North Carolina, US. Molecular prevalence for Anaplasma phagocytophilum was 3% from 68 black bears. No DNA of other Anaplasma or Ehrlichia spp. was identified.

  9. Anaplasma phagocytophilum in horses and ticks in Tunisia

    PubMed Central

    2012-01-01

    Background Anaplasma phagocytophilum , the causative agent of granulocytic anaplasmosis, affects several species of wild and domesticated mammals, including horses. We used direct and indirect methods to compare and evaluate exposure to A. phagocytophilum in horses in northern Tunisia. Methods Serum from 60 horses was tested by IFA for antibodies to A. phagocytophilum , and whole blood was tested for A. phagocytophilum 16S rRNA gene using a nested-PCR. To examine the risk of A. phagocytophilum transmission, 154 ticks that had been collected from horses were examined for the presence of A. phagocytophilum by nested-PCR targeting 16S rRNA gene. Results This is the first time that A. phagocytophilum has been detected in horses in Tunisia, with an overall seroprevalence of 40/60 (67%). Six of the seroreactive samples (10%) had an IFA titer of 1:80, 14 (23%) of 1:160, 8 (13%) of 1:320 and 12 (20%) a titer 1 ≥ 640. The seroprevalence revealed no significant regional and sex differences. In contrast, a significant difference was observed between breeds. Eight (13%) of the horses were positive for A. phagocytophilum in the PCR, with no significant breed and age differences. Hyalomma marginatum was a predominant tick species (130/154), and 3 were infected by A. phagocytophilum (a prevalence of 2.3%). The concordance rate of A. phagocytophilum detection between IFA and PCR had a k value of −0.07. Conclusions The results presented in this study suggest that horses infested by ticks in Tunisia are exposed to A. phagocytophilum. PMID:22935132

  10. Anaplasma phagocytophilum in horses and ticks in Tunisia.

    PubMed

    M'ghirbi, Youmna; Yaïch, Hèla; Ghorbel, Abderazek; Bouattour, Ali

    2012-08-30

    Anaplasma phagocytophilum , the causative agent of granulocytic anaplasmosis, affects several species of wild and domesticated mammals, including horses. We used direct and indirect methods to compare and evaluate exposure to A. phagocytophilum in horses in northern Tunisia. Serum from 60 horses was tested by IFA for antibodies to A. phagocytophilum , and whole blood was tested for A. phagocytophilum 16S rRNA gene using a nested-PCR. To examine the risk of A. phagocytophilum transmission, 154 ticks that had been collected from horses were examined for the presence of A. phagocytophilum by nested-PCR targeting 16S rRNA gene. This is the first time that A. phagocytophilum has been detected in horses in Tunisia, with an overall seroprevalence of 40/60 (67%). Six of the seroreactive samples (10%) had an IFA titer of 1:80, 14 (23%) of 1:160, 8 (13%) of 1:320 and 12 (20%) a titer 1 ≥ 640. The seroprevalence revealed no significant regional and sex differences. In contrast, a significant difference was observed between breeds. Eight (13%) of the horses were positive for A. phagocytophilum in the PCR, with no significant breed and age differences. Hyalomma marginatum was a predominant tick species (130/154), and 3 were infected by A. phagocytophilum (a prevalence of 2.3%). The concordance rate of A. phagocytophilum detection between IFA and PCR had a k value of -0.07. The results presented in this study suggest that horses infested by ticks in Tunisia are exposed to A. phagocytophilum.

  11. Prevalence of Anaplasma phagocytophilum infection and effect on lamb growth

    PubMed Central

    2011-01-01

    Background A major challenge in sheep farming during the grazing season along the coast of south-western Norway is tick-borne fever (TBF) caused by the bacteria Anaplasma phagocytophilum that is transmitted by the tick Ixodes ricinus. Methods A study was carried out in 2007 and 2008 to examine the prevalence of A. phagocytophilum infection and effect on weaning weight in lambs. The study included 1208 lambs from farms in Sunndal Ram Circle in Møre and Romsdal County in Mid-Norway, where ticks are frequently observed. All lambs were blood sampled and serum was analyzed by an indirect fluorescent antibody assay (IFA) to determine an antibody status (positive or negative) to A. phagocytophilum infection. Weight and weight gain and possible effect of infection were analyzed using ANOVA and the MIXED procedure in SAS. Results The overall prevalence of infection with A. phagocytophilum was 55%. A lower weaning weight of 3% (1.34 kg, p < 0.01) was estimated in lambs seropositive to an A. phagocytophilum infection compared to seronegative lambs at an average age of 137 days. Conclusions The results show that A. phagocytophilum infection has an effect on lamb weight gain. The study also support previous findings that A. phagocytophilum infection is widespread in areas where ticks are prevalent, even in flocks treated prophylactic with acaricides. PMID:21569524

  12. Variant -and individual dependent nature of persistent Anaplasma phagocytophilum infection

    PubMed Central

    2010-01-01

    Background Anaplasma phagocytophilum is the causative agent of tick-borne fever in ruminants and human granulocytotropic anaplasmosis (HGA). The bacterium is able to survive for several months in immune-competent sheep by modifying important cellular and humoral defence mechanisms. Little is known about how different strains of A. phagocytophilum propagate in their natural hosts during persistent infection. Methods Two groups of five lambs were infected with each of two 16S rRNA gene variants of A. phagocytophilum, i.e. 16S variant 1 which is identical to GenBank no M73220 and 16S variant 2 which is identical to GenBank no AF336220, respectively. The lambs were infected intravenously and followed by blood sampling for six months. A. phagocytophilum infection in the peripheral blood was detected by absolute quantitative real-time PCR. Results Both 16S rRNA gene variants of A. phagocytophilum established persistent infection for at least six months and showed cyclic bacteraemias, but variant 1 introduced more frequent periods of bacteraemia and higher number of organisms than 16S rRNA gene variant 2 in the peripheral blood. Conclusion Organisms were available from blood more or less constantly during the persistent infection and there were individual differences in cyclic activity of A. phagocytophilum in the infected animals. Two 16S rRNA gene variants of A. phagocytophilum show differences in cyclic activity during persistent infection in lambs. PMID:20398321

  13. Delineating Anaplasma phagocytophilum Ecotypes in Coexisting, Discrete Enzootic Cycles

    PubMed Central

    Bown, Kevin J.; Lambin, Xavier; Ogden, Nicholas H.; Begon, Michael; Telford, Gill; Woldehiwet, Zerai

    2009-01-01

    The emerging tick-borne pathogen Anaplasma phagocytophilum is under increasing scrutiny for the existence of subpopulations that are adapted to different natural cycles. Here, we characterized the diversity of A. phagocytophilum genotypes circulating in a natural system that includes multiple hosts and at least 2 tick species, Ixodes ricinus and the small mammal specialist I. trianguliceps. We encountered numerous genotypes, but only 1 in rodents, with the remainder limited to deer and host-seeking I. ricinus ticks. The absence of the rodent-associated genotype from host-seeking I. ricinus ticks was notable because we demonstrated that rodents fed a large proportion of the I. ricinus larval population and that these larvae were abundant when infections caused by the rodent-associated genotype were prevalent. These observations are consistent with the conclusion that genotypically distinct subpopulations of A. phagocytophilum are restricted to coexisting but separate enzootic cycles and suggest that this restriction may result from specific vector compatibility. PMID:19961674

  14. The pathogen-occupied vacuoles of anaplasma phagocytophilum and anaplasma marginale interact with the endoplasmic reticulum

    USDA-ARS?s Scientific Manuscript database

    The genus Anaplasma consists of tick-transmitted obligate intracellular bacteria that invade white or red blood cells to cause debilitating and potentially fatal infections. A. phagocytophilum, a human and veterinary pathogen, infects neutrophils to cause granulocytic anaplasmosis. A. marginale inva...

  15. [Seroprevalence of Anaplasma phagocytophilum in patients with suspected Lyme borreliosis].

    PubMed

    Dvořáková Heroldová, M; Dvořáčková, M

    2014-11-01

    Human granulocytic anaplasmosis (HGA) is an emerging tick-borne zoonotic disease caused by an obligate intracellular bacterium, Anaplasma phagocytophilum. In Europe, A. phagocytophilum is transmitted by Ixodes ricinus ticks. After Lyme borreliosis and European tick-borne encephalitis, HGA is the third most common tick-borne infection in the USA and Europe. The clinical symptoms of anaplasmosis are non-specific and include malaise, fever, headache, myalgia, and arthralgia. In more severe cases, the gastrointestinal or respiratory tract may be affected. However, most infections are asymptomatic. The aim of our study was to determine the seroprevalence of A. phagocytophilum in patients with suspected Lyme borreliosis. A total of 314 sera from patients with suspected Lyme borreliosis were screened for IgG and IgM antibodies against A. phagocytophilum. The immunoblot assay was used to detect the antibodies. Anti-A. phagocytophilum antibodies were detected in 34 patients, i.e. in 10.82%. IgM antibodies were positive in 19 cases and IgG antibodies in 10 cases. Positivity to both IgM and IgG antibodies was revealed in five patients. Antibodies against Borrelia burgdorferi sensu lato were detected in 181 patients (57.64%). Co-seroprevalence of Borrelia burgdorferi s. l. and A. phagocytophilum was found in 26 patients (8.3%). Positivity for anti-A. phagocytophilum antibodies was most often seen in samples from the age group 60-69 years. Our results show that A. phagocytophilum infection is not uncommon in the Czech Republic and should be considered in patients with a history of a tick bite.

  16. Anaplasma phagocytophilum in central and western Wisconsin: a molecular survey.

    PubMed

    Michalski, M; Rosenfield, C; Erickson, M; Selle, R; Bates, K; Essar, D; Massung, R

    2006-11-01

    Anaplasma phagocytophilum is an obligate intracellular bacterium that is transmitted to humans through the bite of Ixodes spp. ticks, and causes a febrile disease known as human granulocytic anaplasmosis. The presence of A. phagocytophilum in Wisconsin white-tailed deer blood and in deer ticks was assessed using PCR and DNA sequencing. Sampling sites in the western part of the state (Buffalo County) and central region (Waushara, Waupaca, and Green Lake counties) were used. In Buffalo County, 5.6% of deer and 8.9% of ticks were infected. At Hartman Creek State Park (Waupaca County), 11.5% of ticks were infected, while the observed prevalence in deer from counties to the south of the park (Waushara and Green Lake) reached 19-26%. Based on 16S rRNA sequences, A. phagocytophilum strains associated and not associated with human infections were identified. Furthermore, two novel A. phagocytophilum variants were found in deer blood samples. Transmission of Lyme disease has been documented in both the Western and Central regions we sampled, and the presence of A. phagocytophilum in naturally occurring tick populations could present an additional risk of disease to humans that enter tick habitats.

  17. Mechanisms of Obligatory Intracellular Infection with Anaplasma phagocytophilum

    PubMed Central

    Rikihisa, Yasuko

    2011-01-01

    Summary: Anaplasma phagocytophilum persists in nature by cycling between mammals and ticks. Human infection by the bite of an infected tick leads to a potentially fatal emerging disease called human granulocytic anaplasmosis. A. phagocytophilum is an obligatory intracellular bacterium that replicates inside mammalian granulocytes and the salivary gland and midgut cells of ticks. A. phagocytophilum evolved the remarkable ability to hijack the regulatory system of host cells. A. phagocytophilum alters vesicular traffic to create an intracellular membrane-bound compartment that allows replication in seclusion from lysosomes. The bacterium downregulates or actively inhibits a number of innate immune responses of mammalian host cells, and it upregulates cellular cholesterol uptake to acquire cholesterol for survival. It also upregulates several genes critical for the infection of ticks, and it prolongs tick survival at freezing temperatures. Several host factors that exacerbate infection have been identified, including interleukin-8 (IL-8) and cholesterol. Host factors that overcome infection include IL-12 and gamma interferon (IFN-γ). Two bacterial type IV secretion effectors and several bacterial proteins that associate with inclusion membranes have been identified. An understanding of the molecular mechanisms underlying A. phagocytophilum infection will foster the development of creative ideas to prevent or treat this emerging tick-borne disease. PMID:21734244

  18. Anaplasma phagocytophilum infection in moose (Alces alces) in Norway.

    PubMed

    Pūraitė, Irma; Rosef, Olav; Paulauskas, Algimantas; Radzijevskaja, Jana

    2015-01-01

    Anaplasma phagocytophilum is a tick-borne bacterium that infects a wide range of animal species. The aim of our study was to investigate the prevalence of A. phagocytophilum in Norwegian moose Alces alces and to characterize the bacteria by sequencing of partial msp4 and 16S rRNA genes. Hunters collected spleen samples from 99 moose of different ages during 2013 and 2014 in two areas: Aust-Agder County (n = 70) where Ixodes ricinus ticks are abundant and Oppland County (n = 29) where ticks were either absent, or abundance very low. A. phagocytophilum was detected only in moose from the I. ricinus - abundant area. The overall prevalence of infection according to 16S rRNA and msp4 gene-based PCR was 41.4% and 31.4% respectively. Sequence analysis of the partial 16S rRNA and msp4 gene revealed two and eight different sequence types respectively. Four of eight msp4 sequence types determined in this study were unique, while others were identical to sequences derived from other ruminants and ticks. The present study indicates that moose could be a potential wildlife reservoir of A. phagocytophilum in Norway.

  19. Acute transient sensorineural hearing loss due to Anaplasma phagocytophilum.

    PubMed

    Ghera, Princy; Kasirye, Yusuf; Choudhry, Muhammad Waqas; Shaw, Gene R; Ejercito, Victor S

    2011-12-01

    We report the case of a patient who presented with a 3-month history of random epistaxis and recent onset of acute hearing loss associated with fever, chills, and myalgias. Pure tone audiometry revealed bilateral sensorineural hearing loss. Complete blood cell count showed an abnormal neutrophil count of 700/uL (normal >1900/microL) and platelet count of 25 x 10(3)/microL (normal >175 x 10(3)/microL). Giemsa-stained peripheral blood smear revealed neutrophilic intracytoplasmic inclusion consistent with anaplasma morulae. Polymerase chain reaction confirmed Anaplasma phagocytophilum. The patient was treated with oral doxycycline, and, after 14 days of treatment, the hearing loss had improved markedly. Therefore, we concluded that the patient's acute transient bilateral sensorineural hearing loss was associated with anaplasmosis.

  20. Seroprevalence of Anaplasma phagocytophilum in horses in the Czech Republic.

    PubMed

    Praskova, Ivana; Bezdekova, Barbora; Zeman, Petr; Jahn, Petr

    2011-06-01

    Anaplasma phagocytophilum is the aetiological agent of equine granulocytic anaplasmosis (EGA). The aim of this survey was to assess the prevalence of anti-A. phagocytophilum antibodies in the horse population of the Czech Republic (CZ) and to investigate possible links between seropositivity and the geographic origin, age, and/or sex of the tested horses. Antibodies against A. phagocytophilum were screened using an indirect fluorescent antibody method (IFA). Serum samples from 96 healthy horses from 8 localities (7 within the CZ and one control upland/tick-free locality in Ukraine) were examined. While the controls tested negative, the seroprevalence in the Czech localities was estimated at 73%, which suggests that subclinical EGA is common among horses in the CZ. Significant differences between seropositivity rates in individual stables were demonstrated (explicable by the different environment and/or management styles of the horses stabled therein). Horses in stables from which previous manifest cases had been reported tended to have higher average titres. Mares displayed a somewhat higher seropositivity rate than did stallions; age had no demonstrable effect upon the serological status of the examined horses. Copyright © 2011 Elsevier GmbH. All rights reserved.

  1. Prevalence of Anaplasma phagocytophilum in ruminants, rodents and ticks in Gansu, north-western China.

    PubMed

    Yang, Jifei; Liu, Zhijie; Guan, Guiquan; Liu, Qing; Li, Youquan; Chen, Ze; Ma, Miling; Liu, Aihong; Ren, Qiaoyun; Luo, Jianxun; Yin, Hong

    2013-02-01

    The zoonotic rickettsial pathogen Anaplasma phagocytophilum has a broad geographical distribution and a high degree of biological and clinical diversity. To determine the prevalence of Anaplasma phagocytophilum in the Gannan Tibetan Autonomous Prefecture of Gansu Province, north-western China, four ruminant species, one rodent and one tick species were examined for Anaplasma phagocytophilum infection. DNA from Anaplasma phagocytophilum was detected by nested PCR in blood samples from 21/49 sheep (42.9 %), 35/91 goats (38.5 %), 51/158 yaks (32.3 %) and 7/20 cattle-yaks (35.0 %), and in spleen samples from 2/12 rodents (16.7 %). For samples from tick larvae and nymphs, 105 pools were tested; one of 46 larval tick pools was positive and seven of 59 nymphal tick pools were positive. For adult ticks, 40/598 female ticks (6.7 %) and 26/528 male ticks (4.9 %) were positive. The prevalence of Anaplasma phagocytophilum in female ticks was higher than that in males, although the difference was not statistically significant (P>0.05). Sequences analysis based on the 16S rRNA gene indicated that the strains in the study area were distinct from previously reported Anaplasma phagocytophilum in other continents. These results add new information on the epidemiology of Anaplasma phagocytophilum and indicate the tick-animal cycle of anaplasmosis in the area. To the best of our knowledge, this is the first report of Anaplasma phagocytophilum infection in Gansu Province in north-western China.

  2. Anaplasma phagocytophilum in horses and ticks: a preliminary survey of Central Italy.

    PubMed

    Passamonti, Fabrizio; Veronesi, Fabrizia; Cappelli, Katia; Capomaccio, Stefano; Coppola, Giacomo; Marenzoni, Maria Luisa; Piergili, Fioretti Daniela; Verini, Supplizi Andrea; Coletti, Mauro

    2010-01-01

    Anaplasma phagocytophilum, the causative agent of granulocytic ehrlichiosis, affects several species of wild and domesticated mammals, including horses. In this work we compared direct and indirect methods to evaluate A. phagocytophilum presence in Central Italy: 135 sera were screened by IFA for A. phagocytophilum and other haemopathogens (Theileria equi and Babesia caballi). Each horse was also tested for A. phagocytophilum 16S rRNA with a nested-PCR technique. In order to examine the risk of A. phagocytophilum transmission, 114 ticks were examined for the presence of A. phagocytophilum by PCR targeting the 16S rRNA. The seroprevalence against A. phagocytophilum was 17.03% and 11 horses (8.14%) showed positive PCR results. The concordance rate of A. phagocytophilum detection between IFAT and PCR had a K value of 0.34.

  3. A novel high-resolution melt PCR assay discriminates Anaplasma phagocytophilum and "Candidatus Neoehrlichia mikurensis".

    PubMed

    Krücken, Jürgen; Schreiber, Cécile; Maaz, Denny; Kohn, Mareen; Demeler, Janina; Beck, Stefanie; Schein, Eberhard; Olias, Philipp; Richter, Dania; Matuschka, Franz-Rainer; Pachnicke, Stefan; Krieger, Klemens; Kohn, Barbara; von Samson-Himmelstjerna, Georg

    2013-06-01

    "Candidatus Neoehrlichia mikurensis" (Anaplasmataceae) is an emerging pathogen transmitted by Ixodes ticks. Conventional PCR and the newly developed high-resolution melt PCR were used to detect and discriminate "Candidatus Neoehrlichia mikurensis" and Anaplasma phagocytophilum. Both bacterial species were frequently found in Ixodes ricinus and Ixodes hexagonus but virtually absent from Dermacentor reticulatus. In rodents, "Candidatus N. mikurensis" was significantly more prevalent than A. phagocytophilum, whereas in cats, only A. phagocytophilum was found.

  4. The Pathogen-Occupied Vacuoles of Anaplasma phagocytophilum and Anaplasma marginale Interact with the Endoplasmic Reticulum.

    PubMed

    Truchan, Hilary K; Cockburn, Chelsea L; Hebert, Kathryn S; Magunda, Forgivemore; Noh, Susan M; Carlyon, Jason A

    2016-01-01

    The genus Anaplasma consists of tick-transmitted obligate intracellular bacteria that invade white or red blood cells to cause debilitating and potentially fatal infections. A. phagocytophilum, a human and veterinary pathogen, infects neutrophils to cause granulocytic anaplasmosis. A. marginale invades bovine erythrocytes. Evidence suggests that both species may also infect endothelial cells in vivo. In mammalian and arthropod host cells, A. phagocytophilum and A. marginale reside in host cell derived pathogen-occupied vacuoles (POVs). While it was recently demonstrated that the A. phagocytophilum-occupied vacuole (ApV) intercepts membrane traffic from the trans-Golgi network, it is unclear if it or the A. marginale-occupied vacuole (AmV) interacts with other secretory organelles. Here, we demonstrate that the ApV and AmV extensively interact with the host endoplasmic reticulum (ER) in endothelial, myeloid, and/or tick cells. ER lumen markers, calreticulin, and protein disulfide isomerase, and the ER membrane marker, derlin-1, were pronouncedly recruited to the peripheries of both POVs. ApV association with the ER initiated early and continued throughout the infection cycle. Both the ApV and AmV interacted with the rough ER and smooth ER. However, only derlin-1-positive rough ER derived vesicles were delivered into the ApV lumen where they localized with intravacuolar bacteria. Transmission electron microscopy identified multiple ER-POV membrane contact sites on the cytosolic faces of both species' vacuoles that corresponded to areas on the vacuoles' lumenal faces where intravacuolar Anaplasma organisms closely associated. A. phagocytophilum is known to hijack Rab10, a GTPase that regulates ER dynamics and morphology. Yet, ApV-ER interactions were unhindered in cells in which Rab10 had been knocked down, demonstrating that the GTPase is dispensable for the bacterium to parasitize the ER. These data establish the ApV and AmV as pathogen-host interfaces that directly

  5. Draft Anaplasma phagocytophilum Genome Sequences from Five Cows, Two Horses, and One Roe Deer Collected in Europe

    PubMed Central

    Dugat, Thibaud; Rossignol, Marie-Noëlle; Rué, Olivier; Loux, Valentin; Marthey, Sylvain; Moroldo, Marco; Silaghi, Cornelia; Höper, Dirk; Fröhlich, Julia; Pfeffer, Martin; Zweygarth, Erich; Lagrée, Anne-Claire; Boulouis, Henri-Jean

    2016-01-01

    Anaplasma phagocytophilum is a zoonotic tick-borne intracellular bacterium responsible for granulocytic anaplasmosis. As it is difficult to isolate and cultivate, only 20 A. phagocytophilum genomes have been sequenced to date. Here, we present eight A. phagocytophilum genome sequences obtained using alternative approaches based on sequence capture technology. PMID:27908984

  6. Sequence Analysis of the msp4 Gene of Anaplasma phagocytophilum Strains

    PubMed Central

    de la Fuente, José; Massung, Robert F.; Wong, Susan J.; Chu, Frederick K.; Lutz, Hans; Meli, Marina; von Loewenich, Friederike D.; Grzeszczuk, Anna; Torina, Alessandra; Caracappa, Santo; Mangold, Atilio J.; Naranjo, Victoria; Stuen, Snorre; Kocan, Katherine M.

    2005-01-01

    The causative agent of human granulocytic ehrlichiosis was recently reclassified as Anaplasma phagocytophilum, unifying previously described bacteria that cause disease in humans, horses, dogs, and ruminants. For the characterization of genetic heterogeneity in this species, the homologue of Anaplasma marginale major surface protein 4 gene (msp4) was identified, and the coding region was PCR amplified and sequenced from a variety of sources, including 50 samples from the United States, Germany, Poland, Norway, Italy, and Switzerland and 4 samples of A. phagocytophilum-like organisms obtained from white-tailed deer in the United States. Sequence variation between strains of A. phagocytophilum (90 to 100% identity at the nucleotide level and 92 to 100% similarity at the protein level) was higher than in A. marginale. Phylogenetic analyses of msp4 sequences did not provide phylogeographic information but did differentiate strains of A. phagocytophilum obtained from ruminants from those obtained from humans, dogs, and horses. The sequence analysis of the recently discovered A. phagocytophilum msp2 gene corroborated these results. The results reported here suggest that although A. phagocytophilum-like organisms from white-tailed deer may be closely related to A. phagocytophilum, they could be more diverse. These results suggest that A. phagocytophilum strains from ruminants could share some common characteristics, including reservoirs and pathogenicity, which may be different from strains that infect humans. PMID:15750101

  7. The E3 Ubiquitin Ligase XIAP Restricts Anaplasma phagocytophilum Colonization of Ixodes scapularis Ticks

    PubMed Central

    Severo, Maiara S.; Choy, Anthony; Stephens, Kimberly D.; Sakhon, Olivia S.; Chen, Gang; Chung, Duk-Won D.; Le Roch, Karine G.; Blaha, Gregor; Pedra, Joao H. F.

    2013-01-01

    Ubiquitination is a posttranslational modification that regulates protein degradation and signaling in eukaryotes. Although it is acknowledged that pathogens exploit ubiquitination to infect mammalian cells, it remains unknown how microbes interact with the ubiquitination machinery in medically relevant arthropods. Here, we show that the ubiquitination machinery is present in the tick Ixodes scapularis and demonstrate that the E3 ubiquitin ligase named x-linked inhibitor of apoptosis protein (XIAP) restricts bacterial colonization of this arthropod vector. We provide evidence that xiap silencing significantly increases tick colonization by the bacterium Anaplasma phagocytophilum, the causative agent of human granulocytic anaplasmosis. We also demonstrate that (i) XIAP polyubiquitination is dependent on the really interesting new gene (RING) catalytic domain, (ii) XIAP polyubiquitination occurs via lysine (K)-63 but not K-48 residues, and (iii) XIAP-dependent K-63 polyubiquitination requires zinc for catalysis. Taken together, our data define a role for ubiquitination during bacterial colonization of disease vectors. PMID:23901084

  8. Anaplasma phagocytophilum, Babesia microti, and Borrelia burgdorferi in Ixodes scapularis, Southern Coastal Maine

    PubMed Central

    Caporale, Diane A.; Goldberg, John; Lacombe, Eleanor; Lubelczyk, Charles; Rand, Peter W.; Smith, Robert P.

    2004-01-01

    Ixodes scapularis (deer ticks) from Maine were tested for multiple infections by polymerase chain reaction and immunofluorescence. In 1995, 29.5%, 9.5%, and 1.9% of deer ticks were infected with Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti, respectively. In 1996 and 1997, the number of A. phagocytophilum-infected ticks markedly declined. In 1995 through 1996, 4 (1.3%) of 301 were co-infected. PMID:15200875

  9. [First serological study of the prevalence of Anaplasma phagocytophilum in dromedary (Camelus dromedarius) in Tunisia].

    PubMed

    Ben Said, M; Belkahia, H; Sayahi, L; Aloui, M; Jemli, M H; Hadj Mohamed, B; Sassi, L; Darghouth, M A; Djaïem, A A; Bayoudh, M; Messadi, L

    2014-02-01

    The aim of the study was to evaluate the seroprevalence of Anaplasma phagocytophilum in dromedary (Camelus dromedarius). Sera of 226 healthy dromedaries from three regions of Tunisia (Sidi Bouzid, Bouficha and Douz) were tested by indirect immunofluorescence (IFA). The overall infection rate was estimated at 29.2%. The study of risk factors showed that region, age, gender, presence of ticks and types of breeding had no influence on the seroprevalence of A. phagocytophilum. This study indicates for the first time in Tunisia that dromedary may be involved in the natural cycle of A. phagocytophilum.

  10. Molecular evidence of Anaplasma phagocytophilum in wild boar (Sus scrofa) in Belgium

    PubMed Central

    2014-01-01

    Background Anaplasma phagocytophilum is a tick-borne pathogen of veterinary and human importance. Both ticks as vectors and vertebrates as reservoir hosts are essential for the cycle maintenance of this bacterium. Currently, the whole range of animal species reservoirs for A. phagocytophilum in natural environment is still unknown. Therefore, the aim of this study was to estimate the prevalence of infection with A. phagocytophilum in the wild boar population in southern Belgium. Results In the frame of a targeted surveillance program, 513 wild boars were sampled during the hunting season 2011. A nested 16S rRNA PCR was used to screen the presence of A. phagocytophilum DNA in spleen of boars. Within 513 samples, 5 (0,97%) were tested PCR positive and identification was confirmed by sequencing. Conclusions This study gives the first insight of presence of A. phagocytophilum in wild boars in southern Belgium. PMID:24694049

  11. Detection of Borrelia burgdorferi and Anaplasma phagocytophilum in the black-legged tick, Ixodes scapularis, within southwestern Pennsylvania.

    PubMed

    Brown, Scott M; Lehman, Preston M; Kern, Ryan A; Henning, Jill D

    2015-06-01

    Prevalence studies of Borrelia burgdorferi and Anaplasma phagocytophilum have been rare for ticks from southwestern Pennsylvania. We collected 325 Ixodes scapularis ticks between 2011 and 2012 from four counties in southwestern Pennsylvania. We tested for the presence of Borrelia burgdorferi and Anaplasma phagocytophilum using PCR. Of the ticks collected from Pennsylvania, B. burgdorferi (causative agent of Lyme disease) was present in 114/325 (35%) and Anaplasma phagocytophilum (causative agent of Human Granulocytic Anaplasmosis) was present in 48/325 (15%) as determined by PCR analysis.

  12. Premature parturition, edema, and ascites in an alpaca infected with Anaplasma phagocytophilum

    PubMed Central

    Tinkler, Stacy H.; Firshman, Anna M.; Sharkey, Leslie C.

    2012-01-01

    An 8-year-old alpaca was presented for fever, anorexia, edema, ascites, and premature parturition. She was determined to have Anaplasma phagocytophilum infection based on positive blood polymerase chain reaction (PCR) and positive acute and convalescent serum titers. Antibiotics and supportive therapies were administered and the alpaca made a complete recovery. PMID:23633715

  13. Anaplasma phagocytophilum infection (granulocytic anaplasmosis) in a dog from Vancouver Island

    PubMed Central

    2005-01-01

    Abstract A 7-year-old Labrador retriever had nonspecific clinical signs that included lethargy, malaise, and difficult ambulation. The dog was native to Vancouver Island, British Columbia, and had never left this area. Morulae were identified in polymorphonuclear cells. Serologic studies and polymerase chain reaction (PCR) testing confirmed canine anaplasmosis caused by Anaplasma phagocytophilum. The dog recovered after treatment with tetracycline. PMID:16231653

  14. First molecular detection and genetic analysis of Anaplasma phagocytophilum in shelter cats in Seoul, Korea.

    PubMed

    Lee, Seung-Hun; VanBik, Dorene; Kim, Neung-Hee; Park, Sang-Joon; Kwon, Oh-Deog; Kim, Tae-Hwan; Kwak, Dongmi

    2016-12-01

    Here, we report the molecular detection of Anaplasma phagocytophilum in shelter cats in Korea and the relationships between A. phagocytophilum gene sequences and the pathogenicity, region, and host specificity of this bacterium. Two (0.9%) out of 222 shelter cats from Seoul, Korea, yielded positive results for the A. phagocytophilum 16S rRNA, groEL, and msp2 genes. Phylogenetic analysis divided groEL gene sequences into two groups (alanine and serine), based on their nucleotide and amino acid sequences. A. phagocytophilum msp2 gene sequences were grouped per the region of isolation (Europe vs. USA, including Korea). Some nucleotide and amino acid sequences of groEL and msp2 showed distinctive patterns according to the region of isolation, which helped in distinguishing A. phagocytophilum gene sequences detected in Korea from those detected in the USA and Europe. Although the limited number of clinical anaplasmosis cases caused by A. phagocytophilum belonging to the alanine group prevents any firm conclusions, the results of the present study tend to refute the previous view that the pathogenicity of A. phagocytophilum is associated with the serine group. Moreover, our results suggest that genetic analyses of groEL and msp2 can be used to obtain a regional fingerprint of A. phagocytophilum.

  15. First molecular detection and genetic analysis of Anaplasma phagocytophilum in shelter cats in Seoul, Korea.

    PubMed

    Lee, Seung-Hun; VanBik, Dorene; Kim, Neung-Hee; Park, Sang-Joon; Kwon, Oh-Deog; Kim, Tae-Hwan; Kwak, Dongmi

    2016-10-29

    Here, we report the molecular detection of Anaplasma phagocytophilum in shelter cats in Korea and the relationships between A. phagocytophilum gene sequences and the pathogenicity, region, and host specificity of this bacterium. Two (0.9%) out of 222 shelter cats from Seoul, Korea, yielded positive results for the A. phagocytophilum 16S rRNA, groEL, and msp2 genes. Phylogenetic analysis divided groEL gene sequences into two groups (alanine and serine), based on their nucleotide and amino acid sequences. A. phagocytophilum msp2 gene sequences were grouped per the region of isolation (Europe vs. USA, including Korea). Some nucleotide and amino acid sequences of groEL and msp2 showed distinctive patterns according to the region of isolation, which helped in distinguishing A. phagocytophilum gene sequences detected in Korea from those detected in the USA and Europe. Although the limited number of clinical anaplasmosis cases caused by A. phagocytophilum belonging to the alanine group prevents any firm conclusions, the results of the present study tend to refute the previous view that the pathogenicity of A. phagocytophilum is associated with the serine group. Moreover, our results suggest that genetic analyses of groEL and msp2 can be used to obtain a regional fingerprint of A. phagocytophilum.

  16. Molecular surveillance of Theileria equi and Anaplasma phagocytophilum infections in horses from Ukraine, Poland and Slovakia.

    PubMed

    Slivinska, Kateryna; Víchová, Bronislava; Werszko, Joanna; Szewczyk, Tomasz; Wróblewski, Zbigniew; Peťko, Branislav; Ragač, Ondrej; Demeshkant, Vitaliy; Karbowiak, Grzegorz

    2016-01-15

    A survey was undertaken to assess the prevalence of Theileria equi and Anaplasma phagocytophilum in some regions of Ukraine, Poland and Slovakia. Using a specific PCR assays, blood samples from 215 horses were tested. The prevalence of T. equi and A. phagocytophilum infection was 13.95% and 1.4%, respectively. BLAST analysis showed the isolates closest to the T. equi 18S rRNA and A. phagocytophilum msp4 gene sequences in GenBank with a similarity of ≥99%. No significant association was found between the T. equi PCR positivity and the age or sex of the horses. There was a significant association between the origin of horses and T. equi-PCR positivity. No significant association was found between the A. phagocytophilum-PCR positivity and the age, sex or origin. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Prevalence of Anaplasma phagocytophilum in fallow deer (Dama dama) and feeding ticks from an Italy preserve.

    PubMed

    Veronesi, F; Galuppi, R; Tampieri, M P; Bonoli, C; Mammoli, R; Piergili Fioretti, D

    2011-02-01

    Up to date, information concerning the Anaplasma phagocytophilum infection in fallow deer is scant, therefore, to verify its prevalence in these ungulates serological and PCR screenings were performed on blood of 72 fallow deer hunted in a Central-Northern Italian preserve. Molecular analyses were also performed on 90 ticks removed from the animals. A. phagocytophilum infection in fallow deer was confirmed in 20 out 72 by IFA assay and in 11 out 72 by PCR. The sequence obtained revealed a complete genetic homology among the blood samples and strong degrees of homology with other European isolates. Considering the 90 ticks collected we found that 7.3% of Ixodes ricinus harboured A. phagocytophilum specific DNA. The data obtained confirmed that fallow deer can be a competent host for A. phagocytophilum and, therefore, that may represent a biological reservoir playing an important role in the epidemiological scenarios of the infection, in the geographical areas where is widespread.

  18. Comparative genomics of first available bovine Anaplasma phagocytophilum genome obtained with targeted sequence capture.

    PubMed

    Dugat, Thibaud; Loux, Valentin; Marthey, Sylvain; Moroldo, Marco; Lagrée, Anne-Claire; Boulouis, Henri-Jean; Haddad, Nadia; Maillard, Renaud

    2014-11-17

    Anaplasma phagocytophilum is a zoonotic and obligate intracellular bacterium transmitted by ticks. In domestic ruminants, it is the causative agent of tick-borne fever, which causes significant economic losses in Europe. As A. phagocytophilum is difficult to isolate and cultivate, only nine genome sequences have been published to date, none of which originate from a bovine strain.Our goals were to; 1/ develop a sequencing methodology which efficiently circumvents the difficulties associated with A. phagocytophilum isolation and culture; 2/ describe the first genome of a bovine strain; and 3/ compare it with available genomes, in order to both explore key genomic features at the species level, and to identify candidate genes that could be specific to bovine strains. DNA was extracted from a bovine blood sample infected by A. phagocytophilum. Following a whole genome capture approach, A. phagocytophilum DNA was enriched 197-fold in the sample and then sequenced using Illumina technology. In total, 58.9% of obtained reads corresponded to the A. phagocytophilum genome, covering 85.3% of the HZ genome. Then by performing comparisons with nine previously-sequenced A. phagocytophilum genomes, we determined the core genome of these ten strains. Following analysis, 1281 coding DNA sequences, including 1001 complete sequences, were detected in the A. phagocytophilum bovine genome, of which four appeared to be unique to the bovine isolate. These four coding DNA sequences coded for "hypothetical proteins of unknown function" and require further analysis. We also identified nine proteins common to both European domestic ruminants tested. Using a whole genome capture approach, we have sequenced the first A. phagocytophilum genome isolated from a cow. To the best of our knowledge, this is the first time that this method has been used to selectively enrich pathogenic bacterial DNA from samples also containing host DNA. The four proteins unique to the A. phagocytophilum bovine

  19. Anaplasma phagocytophilum infection modulates expression of megakaryocyte cell cycle genes through phosphatidylinositol-3-kinase signaling.

    PubMed

    Khanal, Supreet; Sultana, Hameeda; Catravas, John D; Carlyon, Jason A; Neelakanta, Girish

    2017-01-01

    Anaplasma phagocytophilum, the agent of human granulocytic anaplasmosis infects neutrophils and other cells from hematopoietic origin. Using human megakaryocytic cell line, MEG-01, we show that expression of cell cycle genes in these cells are altered upon A. phagocytophilum infection. Expression of several cell cycle genes in MEG-01 cells was significantly up regulated at early and then down regulated at later stages of A. phagocytophilum infection. Lactate dehydrogenase (LDH) assays revealed reduced cellular cytotoxicity in MEG-01 cells upon A. phagocytophilum infection. The levels of both PI3KCA (p110 alpha, catalytic subunit) and PI3KR1 (p85, regulatory subunit) of Class I PI3 kinases and phosphorylated protein kinase B (Akt/PKB) and inhibitory kappa B (IκB) were elevated at both early and late stages of A. phagocytophilum infection. Inhibition of PI3 kinases with LY294002 treatment resulted in significant reduction in the expression of tested cell cycle genes, A. phagocytophilum burden and phosphorylated Akt levels in these MEG-01 cells. Collectively, these results suggest a role for PI3K-Akt-NF-κB signaling pathway in the modulation of megakaryocyte cell cycle genes upon A. phagocytophilum infection.

  20. Opening the black box of Anaplasma phagocytophilum diversity: current situation and future perspectives

    PubMed Central

    Dugat, Thibaud; Lagrée, Anne-Claire; Maillard, Renaud; Boulouis, Henri-Jean; Haddad, Nadia

    2015-01-01

    Anaplasma phagocytophilum is a zoonotic obligate intracellular bacterium known to be transmitted by ticks belonging to the Ixodes persulcatus complex. This bacterium can infect several mammalian species, and is known to cause diseases with variable symptoms in many domestic animals. Specifically, it is the causative agent of tick-borne fever (TBF), a disease of important economic impact in European domestic ruminants, and human granulocytic anaplasmosis (HGA), an emerging zoonotic disease in Asia, USA and Europe. A. phagocytophilum epidemiological cycles are complex and involve different ecotypes, vectors, and mammalian host species. Moreover, the epidemiology of A. phagocytophilum infection differs greatly between Europe and the USA. These different epidemiological contexts are associated with considerable variations in bacterial strains. Until recently, few A. phagocytophilum molecular typing tools were available, generating difficulties in completely elucidating the epidemiological cycles of this bacterium. Over the last few years, many A. phagocytophilum typing techniques have been developed, permitting in-depth epidemiological exploration. Here, we review the current knowledge and future perspectives regarding A. phagocytophilum epidemiology and phylogeny, and then focus on the molecular typing tools available for studying A. phagocytophilum genetic diversity. PMID:26322277

  1. The prenylation inhibitor manumycin A reduces the viability of Anaplasma phagocytophilum

    PubMed Central

    Xiong, Qingming

    2011-01-01

    Anaplasma phagocytophilum is an obligately intracellular bacterium and is the causative agent of human granulocytic anaplasmosis (HGA), an emerging and major tick-borne disease in the USA and other parts of the world. This study showed that the prenylation inhibitor manumycin A effectively blocked A. phagocytophilum infection in host cells (HL-60 or RF/6A cells). A. phagocytophilum infection activated extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase in host cells, and manumycin A treatment reduced ERK activation in A. phagocytophilum-infected host cells. As ERK activation is required for A. phagocytophilum infection, we examined whether manumycin A inhibited the bacteria directly or through host ERK signalling. Treatment of A. phagocytophilum alone with manumycin A significantly reduced the bacterial infectivity of host cells and bacterial viability in the absence of host cells, whereas pre-treatment of host cells did not inhibit bacterial infection in host cells. The inhibitory effect of manumycin A on A. phagocytophilum infection in host cells was achieved even at a concentration 100 times lower than that required for effective inhibition of mammalian cell signalling. These results suggested that manumycin A directly inactivates the bacterium, resulting in reduced infection and ERK1/2 activation. Thus, the manumycin group of drugs may have a therapeutic potential for HGA. PMID:21349982

  2. Novel Genetic Variants of Anaplasma phagocytophilum, Anaplasma bovis, Anaplasma centrale, and a Novel Ehrlichia sp. in Wild Deer and Ticks on Two Major Islands in Japan

    PubMed Central

    Kawahara, Makoto; Rikihisa, Yasuko; Lin, Quan; Isogai, Emiko; Tahara, Kenji; Itagaki, Asao; Hiramitsu, Yoshimichi; Tajima, Tomoko

    2006-01-01

    Wild deer are one of the important natural reservoir hosts of several species of Ehrlichia and Anaplasma that cause human ehrlichiosis or anaplasmosis in the United States and Europe. The primary aim of the present study was to determine whether and what species of Ehrlichia and Anaplasma naturally infect deer in Japan. Blood samples obtained from wild deer on two major Japanese islands, Hokkaido and Honshu, were tested for the presence of Ehrlichia and Anaplasma by PCR assays and sequencing of the 16S rRNA genes, major outer membrane protein p44 genes, and groESL. DNA representing four species and two genera of Ehrlichia and Anaplasma was identified in 33 of 126 wild deer (26%). DNA sequence analysis revealed novel strains of Anaplasma phagocytophilum, a novel Ehrlichia sp., Anaplasma centrale, and Anaplasma bovis in the blood samples from deer. None of these have been found previously in deer. The new Ehrlichia sp., A. bovis, and A. centrale were also detected in Hemaphysalis longicornis ticks from Honshu Island. These results suggest that enzootic cycles of Ehrlichia and Anaplasma species distinct from those found in the United States or Europe have been established in wild deer and ticks in Japan. PMID:16461655

  3. Detection of a non-pathogenic variant of Anaplasma phagocytophilum in Ixodes ricinus from La Rioja, Spain.

    PubMed

    Portillo, A; Santos, A S; Santibáñez, S; Pérez-Martínez, L; Blanco, J R; Ibarra, V; Oteo, J A

    2005-12-01

    Our aim was to identify variants of Anaplasma phagocytophilum 16S rRNA gene sequences among products amplified from Ixodes ricinus collected in La Rioja, Spain. A. phagocytophilum AP-variant 1, reported as non-pathogenic, was detected in 12 samples (two adults and ten nymphs). This finding could justify the low incidence of human anaplasmosis in our area, despite the high prevalence of A. phagocytophilum in ticks.

  4. Serological evidence of exposure to Ehrlichia canis and Anaplasma phagocytophilum in Central Italian healthy domestic cats.

    PubMed

    Ebani, Valentina V; Bertelloni, Fabrizio

    2014-10-01

    The aim of the present survey was to estimate the seroprevalences of Ehrlichia canis and Anaplasma phagocytophilum in the Central Italian feline population. Serum samples of 560 healthy domestic cats were examined by indirect immunofluorescence assay (IFAT), considering an antibody titre of 1:40 as cut-off. Seroprevalences of 6.4% and 4.5% were found for E. canis and A. phagocytophilum, respectively. Adult, mixed breed cats showed seroprevalences higher than younger and purebred subjects, whereas no differences were observed in relation to gender and living conditions.

  5. Molecular survey of Anaplasma phagocytophilum and Ehrlichia canis in red foxes (Vulpes vulpes) from central Italy.

    PubMed

    Ebani, Valentina Virginia; Verin, Ranieri; Fratini, Filippo; Poli, Alessandro; Cerri, Domenico

    2011-07-01

    During the 2007-2008 hunting season, 150 spleen samples were collected from free-ranging red foxes (Vulpes vulpes) in central Italy. The specimens were tested by two nested PCR assays to detect DNA of Anaplasma phagocytophilum, etiologic agent of granulocytic ehrlichiosis of animals and humans, and DNA of Ehrlichia canis, which causes the monocytic ehrlichiosis in canids. None of the foxes were PCR-positive for E. canis; 25 (16.6%) were positive for A. phagocytophilum. No specific gross alterations were detected at necropsy, and no histopathologic lesions found on PCR-positive spleen samples.

  6. Anaplasma phagocytophilum variants in sympatric red deer (Cervus elaphus) and sheep in southern Norway.

    PubMed

    Stuen, Snorre; Pettersen, Kristin Sæbø; Granquist, Erik Georg; Bergström, Karin; Bown, Kevin John; Birtles, Richard John

    2013-04-01

    Infections by the ixodid tick-transmitted bacterial pathogen Anaplasma phagocytophilum are common in domestic ruminants and cervids in the coastal areas of southern Norway. Previous experimental work has shown that A. phagocytophilum strains recovered from red deer (Cervus elaphus) are infective in lambs, but epidemiological links between infections in red deer and sheep have yet to be established. To address this shortfall, the present study explores the genotypic relatedness between A. phagocytophilum strains infecting sympatric red deer and sheep. Blood from 32 lambs grazing on tick-infested pasture, and blood and tissues from 8 red deer shot in proximity to these pastures were collected during the summer and autumn of 2007. The presence of A. phagocytophilum in these samples was determined by PCR-based methods, and genotyping of detected strains was performed using comparative sequence analysis of 16S rDNA and msp4 fragments. A. phagocytophilum DNA was detected in 12 lambs and 7 red deer, 11 and 4 individuals of which 16S rDNA and msp4 sequence data were obtained from, respectively. A total of 9 genotypes were delineated, and only different individuals of the same host species were infected with indistinguishable A. phagocytophilum genotypes. Although 3 of the red deer-infecting genotypes belonged to a cluster of exclusively deer-associated strains phylogenetically remote from those commonly encountered in sheep, one red deer-infecting genotype, although unique, clustered tightly with genotypes associated with a wide range of hosts including sheep.

  7. Molecular analysis of Anaplasma phagocytophilum and Babesia divergens in red deer (Cervus elaphus) in Western Austria.

    PubMed

    Cézanne, Rita; Mrowietz, Naike; Eigner, Barbara; Duscher, Georg Gerhard; Glawischnig, Walter; Fuehrer, Hans-Peter

    2017-02-01

    Wild ungulates may act as reservoirs of various vector borne pathogens that can infect humans and domestic animals. In the present study, blood samples from 196 red deer (Cervus elaphus) from Western Austria (Vorarlberg, Tyrol and Salzburg) were collected on filter paper and tested for Anaplasmataceae, Piroplasmida, Rickettsia and filarioid helminths using molecular tools. Babesia divergens was detected in ten (5.1%) and Anaplasma phagocytophilum in three (1.5%) of the 196 samples. Filarioid helminths, Rickettsia spp. and Theileria spp. were not detected. These findings indicate that red deer may serve as reservoirs of Babesia divergens and Anaplasma phagocytophilum in Western Austria. Further investigations are needed to assess the presence of these pathogens in ticks in this geographical region, and the significance of these pathogens in both animals and humans.

  8. Anaplasma phagocytophilum infection in a horse from Switzerland with severe neurological symptoms.

    PubMed

    Gussmann, K; Czech, C; Hermann, M; Schaarschmidt-Kiener, D; von Loewenich, F D

    2014-07-01

    A 22-year old mare from Switzerland was admitted to an equine clinic in May 2011. She presented with fever, lethargy, icteric mucous membranes, reduced alertness, an unsteady gait and ataxia. An Anaplasma phagocytophilum infection was confirmed by blood smear and PCR. The mare was treated with oxytetracylin and recovered rapidly, but she still suffered from a slight atactic gait disturbance at 3 weeks post infection.

  9. Detection of Anaplasma phagocytophilum DNA in Ixodes Ticks (Acari: Ixodidae) from Madeira Island and Setúbal District, Mainland Portugal

    PubMed Central

    Santos-Silva, Maria Margarida; Almeida, Victor Carlos; Bacellar, Fátima; Dumler, John Stephen

    2004-01-01

    A total of 278 Ixodes ticks, collected from Madeira Island and Setúbal District, mainland Portugal, were examined by polymerase chain reaction (PCR) for the presence of Anaplasma phagocytophilum. Six (4%) of 142 Ixodes ricinus nymphs collected in Madeira Island and 1 nymph and 1 male (2%) of 93 I. ventalloi collected in Setúbal District tested positive for A. phagocytophilum msp2 genes or rrs. Infection was not detected among 43 I. ricinus on mainland Portugal. All PCR products were confirmed by nucleotide sequencing to be identical or to be most closely related to A. phagocytophilum. To our knowledge, this is the first evidence of A. phagocytophilum in ticks from Setúbal District, mainland Portugal, and the first documentation of Anaplasma infection in I. ventalloi. Moreover, these findings confirm the persistence of A. phagocytophilum in Madeira Island's I. ricinus. PMID:15498168

  10. Detection of Anaplasma phagocytophilum DNA in Ixodes ticks (Acari: Ixodidae) from Madeira Island and Setubal District, mainland Portugal.

    PubMed

    Santos, Ana Sofia; Santos-Silva, Maria Margarida; Almeida, Victor Carlos; Bacellar, Fátima; Dumler, John Stephen

    2004-09-01

    A total of 278 Ixodes ticks, collected from Madeira Island and Setubal District, mainland Portugal, were examined by polymerase chain reaction (PCR) for the presence of Anaplasma phagocytophilum. Six (4%) of 142 Ixodes ricinus nymphs collected in Madeira Island and 1 nymph and 1 male (2%) of 93 I. ventalloi collected in Setubal District tested positive for A. phagocytophilum msp2 genes or rrs. Infection was not detected among 43 I. ricinus on mainland Portugal. All PCR products were confirmed by nucleotide sequencing to be identical or to be most closely related to A. phagocytophilum. To our knowledge, this is the first evidence of A. phagocytophilum in ticks from Setubal District, mainland Portugal, and the first documentation of Anaplasma infection in I. ventalloi. Moreover, these findings confirm the persistence of A. phagocytophilum in Madeira Island's I. ricinus.

  11. Anaplasma phagocytophilum in questing Ixodes ricinus ticks from Romania.

    PubMed

    Matei, Ioana Adriana; Kalmár, Zsuzsa; Magdaş, Cristian; Magdaş, Virginia; Toriay, Hortenzia; Dumitrache, Mirabela Oana; Ionică, Angela Monica; D'Amico, Gianluca; Sándor, Attila D; Mărcuţan, Daniel Ioan; Domşa, Cristian; Gherman, Călin Mircea; Mihalca, Andrei Daniel

    2015-04-01

    Granulocytic anaplasmosis is a common vector-borne disease of humans and animals with natural transmission cycle that involves tick vectors, among which Ixodes ricinus is the most important. The present paper reports the prevalence and geographical distribution of A. phagocytophilum in 10,438 questing Ixodes ricinus ticks collected at 113 locations from 40 counties of Romania. The unfed ticks were examined for the presence of A. phagocytophilum by PCR targeting a portion of ankA gene. The overall prevalence of infection was 3.42%, with local prevalences ranging between 0.29% and 22.45%, with an average prevalence of 5.39% in the infected localities. The infection with A. phagocytophilum was detected in 72 out of 113 localities and in 34 out of 40 counties. The highest prevalence was recorded in females followed by males and nymphs. The results and the distribution model have shown a large distribution of A. phagocytophilum, covering Romania's entire territory. This study is the first large scale survey of the presence of A. phagocytophilum in questing I. ricinus ticks from Romania.

  12. Integrated Metabolomics, Transcriptomics and Proteomics Identifies Metabolic Pathways Affected by Anaplasma phagocytophilum Infection in Tick Cells*

    PubMed Central

    Villar, Margarita; Ayllón, Nieves; Alberdi, Pilar; Moreno, Andrés; Moreno, María; Tobes, Raquel; Mateos-Hernández, Lourdes; Weisheit, Sabine; Bell-Sakyi, Lesley; de la Fuente, José

    2015-01-01

    Anaplasma phagocytophilum is an emerging zoonotic pathogen that causes human granulocytic anaplasmosis. These intracellular bacteria establish infection by affecting cell function in both the vertebrate host and the tick vector, Ixodes scapularis. Previous studies have characterized the tick transcriptome and proteome in response to A. phagocytophilum infection. However, in the postgenomic era, the integration of omics datasets through a systems biology approach allows network-based analyses to describe the complexity and functionality of biological systems such as host–pathogen interactions and the discovery of new targets for prevention and control of infectious diseases. This study reports the first systems biology integration of metabolomics, transcriptomics, and proteomics data to characterize essential metabolic pathways involved in the tick response to A. phagocytophilum infection. The ISE6 tick cells used in this study constitute a model for hemocytes involved in pathogen infection and immune response. The results showed that infection affected protein processing in endoplasmic reticulum and glucose metabolic pathways in tick cells. These results supported tick–Anaplasma co-evolution by providing new evidence of how tick cells limit pathogen infection, while the pathogen benefits from the tick cell response to establish infection. Additionally, ticks benefit from A. phagocytophilum infection by increasing survival while pathogens guarantee transmission. The results suggested that A. phagocytophilum induces protein misfolding to limit the tick cell response and facilitate infection but requires protein degradation to prevent ER stress and cell apoptosis to survive in infected cells. Additionally, A. phagocytophilum may benefit from the tick cell's ability to limit bacterial infection through PEPCK inhibition leading to decreased glucose metabolism, which also results in the inhibition of cell apoptosis that increases infection of tick cells. These

  13. Integrated Metabolomics, Transcriptomics and Proteomics Identifies Metabolic Pathways Affected by Anaplasma phagocytophilum Infection in Tick Cells.

    PubMed

    Villar, Margarita; Ayllón, Nieves; Alberdi, Pilar; Moreno, Andrés; Moreno, María; Tobes, Raquel; Mateos-Hernández, Lourdes; Weisheit, Sabine; Bell-Sakyi, Lesley; de la Fuente, José

    2015-12-01

    Anaplasma phagocytophilum is an emerging zoonotic pathogen that causes human granulocytic anaplasmosis. These intracellular bacteria establish infection by affecting cell function in both the vertebrate host and the tick vector, Ixodes scapularis. Previous studies have characterized the tick transcriptome and proteome in response to A. phagocytophilum infection. However, in the postgenomic era, the integration of omics datasets through a systems biology approach allows network-based analyses to describe the complexity and functionality of biological systems such as host-pathogen interactions and the discovery of new targets for prevention and control of infectious diseases. This study reports the first systems biology integration of metabolomics, transcriptomics, and proteomics data to characterize essential metabolic pathways involved in the tick response to A. phagocytophilum infection. The ISE6 tick cells used in this study constitute a model for hemocytes involved in pathogen infection and immune response. The results showed that infection affected protein processing in endoplasmic reticulum and glucose metabolic pathways in tick cells. These results supported tick-Anaplasma co-evolution by providing new evidence of how tick cells limit pathogen infection, while the pathogen benefits from the tick cell response to establish infection. Additionally, ticks benefit from A. phagocytophilum infection by increasing survival while pathogens guarantee transmission. The results suggested that A. phagocytophilum induces protein misfolding to limit the tick cell response and facilitate infection but requires protein degradation to prevent ER stress and cell apoptosis to survive in infected cells. Additionally, A. phagocytophilum may benefit from the tick cell's ability to limit bacterial infection through PEPCK inhibition leading to decreased glucose metabolism, which also results in the inhibition of cell apoptosis that increases infection of tick cells. These results

  14. Identification and Characterization of Anaplasma phagocytophilum Proteins Involved in Infection of the Tick Vector, Ixodes scapularis

    PubMed Central

    Kocan, Katherine M.; Bonzón-Kulichenko, Elena; Alberdi, Pilar; Blouin, Edmour F.; Weisheit, Sabine; Mateos-Hernández, Lourdes; Cabezas-Cruz, Alejandro; Bell-Sakyi, Lesley; Vancová, Marie; Bílý, Tomáš; Meyer, Damien F.; Sterba, Jan; Contreras, Marinela; Rudenko, Nataliia; Grubhoffer, Libor; Vázquez, Jesús; de la Fuente, José

    2015-01-01

    Anaplasma phagocytophilum is an emerging zoonotic pathogen transmitted by Ixodes scapularis that causes human granulocytic anaplasmosis. Here, a high throughput quantitative proteomics approach was used to characterize A. phagocytophilum proteome during rickettsial multiplication and identify proteins involved in infection of the tick vector, I. scapularis. The first step in this research was focused on tick cells infected with A. phagocytophilum and sampled at two time points containing 10–15% and 65–71% infected cells, respectively to identify key bacterial proteins over-represented in high percentage infected cells. The second step was focused on adult female tick guts and salivary glands infected with A. phagocytophilum to compare in vitro results with those occurring during bacterial infection in vivo. The results showed differences in the proteome of A. phagocytophilum in infected ticks with higher impact on protein synthesis and processing than on bacterial replication in tick salivary glands. These results correlated well with the developmental cycle of A. phagocytophilum, in which cells convert from an intracellular reticulated, replicative form to the nondividing infectious dense-core form. The analysis of A. phagocytophilum differentially represented proteins identified stress response (GroEL, HSP70) and surface (MSP4) proteins that were over-represented in high percentage infected tick cells and salivary glands when compared to low percentage infected cells and guts, respectively. The results demonstrated that MSP4, GroEL and HSP70 interact and bind to tick cells, thus playing a role in rickettsia-tick interactions. The most important finding of these studies is the increase in the level of certain bacterial stress response and surface proteins in A. phagocytophilum-infected tick cells and salivary glands with functional implication in tick-pathogen interactions. These results gave a new dimension to the role of these stress response and surface

  15. Development of a new PCR-based assay to detect Anaplasmataceae and the first report of Anaplasma phagocytophilum and Anaplasma platys in cattle from Algeria.

    PubMed

    Dahmani, Mustapha; Davoust, Bernard; Benterki, Mohamed Seghir; Fenollar, Florence; Raoult, Didier; Mediannikov, Oleg

    2015-04-01

    Bovine anaplasmosis is a hemoparasitic disease considered as a major constraint to cattle production in many countries. This pathology is at least partially caused by Anaplasma phagocytophilum, Anaplasma marginale, Anaplasma centrale, and Anaplasma bovis. The global threat and emergence of these species in animals require the reliable identification of these bacteria in animal samples. In this study, we developed a new qPCR tool targeting the 23S rRNA gene for the detection of Anaplasmataceae bacteria. The primers and probe for the qPCR reaction had 100% specificity and could identify at least A. phagocytophilum, A. marginale, A. centrale, Anaplasma ovis, Anaplasma platys, Ehrlichia canis, Ehrlichia ruminantium, Neorickettisa sennetsu, and Neorickettsia risticii. We used this tool to test samples of bovines from Batna (Algeria), an area from which bovine anaplasmosis have never been reported. We identified three genetic variants of A. phagocytophilum, A. platys and Anaplasma sp. "variant 4". This finding should attract the attention of public authorities to assess the involvement of these pathogens in human and animal health.

  16. Subversion of NPC1 pathway of cholesterol transport by Anaplasma phagocytophilum

    PubMed Central

    Xiong, Qingming; Rikihisa, Yasuko

    2013-01-01

    Summary Intracellular cholesterol amounts, distribution, and traffic are tightly regulated to maintain the healthy eukaryotic cell function. However, how intracellular pathogens that require cholesterol, interact with the host cholesterol homeostasis and traffic is not well understood. Anaplasma phagocytophilum is an obligatory intracellular and cholesterol-robbing bacterium, which causes human granulocytic anaplasmosis. Here we found that a subset of cholesterol-binding membrane protein, Niemann–Pick type C1 (NPC1)-bearing vesicles devoid of lysosomal markers were upregulated in HL-60 cells infected with A. phagocytophilum, and trafficked to live bacterial inclusions. The NPC1 localization to A. phagocytophilum inclusions was abolished by low-density lipoprotein (LDL)-derived cholesterol traffic inhibitor U18666A. Studies using NPC1 siRNA and the cell line with cholesterol traffic defect demonstrated that the NPC1 function is required for bacterial cholesterol acquisition and infection. Furthermore, trans-Golgi network-specific soluble N-ethylmaleimide-sensitive factor attachment protein receptors, vesicle-associated membrane protein (VAMP4) and syntaxin 16, which are associated with NPC1 and LDL-derived cholesterol vesicular transport were recruited to A. phagocytophilum inclusions, and VAMP4 was required for bacteria infection. Taken together, A. phagocytophilum is the first example of a pathogen that subverts the NPC1 pathway of intracellular cholesterol transport and homeostasis for bacterial inclusion membrane biogenesis and cholesterol capture. PMID:22212234

  17. Anaplasma phagocytophilum strains from voles and shrews exhibit specific ankA gene sequences

    PubMed Central

    2013-01-01

    Background Anaplasma phagocytophilum is a Gram-negative bacterium that replicates obligate intracellularly in neutrophils. It is transmitted by Ixodes spp. ticks and causes acute febrile disease in humans, dogs, horses, cats, and livestock. Because A. phagocytophilum is not transmitted transovarially in Ixodes spp., it is thought to depend on reservoir hosts to complete its life cycle. In Europe, A. phagocytophilum was detected in roe deer, red deer, wild boars, and small mammals. In contrast to roe deer, red deer and wild boars have been considered as reservoir hosts for granulocytic anaplasmosis in humans, dogs, and horses according to groESL- and ankA-based genotyping. A. phagocytophilum variants infecting small mammals in Europe have not been characterized extensively to date. Results We amplified the total ankA open reading frames of 27 strains from voles and shrews. The analysis revealed that they harboured A. phagocytophilum strains that belonged to a distinct newly described ankA gene cluster. Further, we provide evidence that the heterogeneity of ankA gene sequences might have arisen via recombination. Conclusions Based on ankA-based genotyping voles and shrews are unlikely reservoir hosts for granulocytic anaplasmosis in humans, dogs, horses, and livestock in Europe. PMID:24283328

  18. Global proteomic analysis of two tick-borne emerging zoonotic agents: Anaplasma phagocytophilum and Ehrlichia chaffeensis

    SciTech Connect

    Lin, Mingqun ..; Kikuchi, Takane; Brewer, Heather M.; Norbeck, Angela D.; Rikihisa, Yasuko

    2011-02-17

    Anaplasma phagocytophilum and Ehrlichia chaffeensis are obligatory intracellular {alpha}-proteobacteria that infect human leukocytes and cause potentially fatal emerging zoonoses. In the present study, we determined global protein expression profiles of these bacteria cultured in the human promyelocytic leukemia cell line, HL-60. Mass spectrometric (MS) analyses identified a total of 1,212 A. phagocytophilum and 1,021 E. chaffeensis proteins, representing 89.3 and 92.3% of the predicted bacterial proteomes, respectively. Nearly all bacterial proteins ({approx}99%) with known functions were expressed, whereas only approximately 80% of hypothetical proteins were detected in infected human cells. Quantitative MS/MS analyses indicated that highly expressed proteins in both bacteria included chaperones, enzymes involved in biosynthesis and metabolism, and outer membrane proteins, such as A. phagocytophilum P44 and E. chaffeensis P28/OMP-1. Among 113 A. phagocytophilum p44 paralogous genes, 110 of them were expressed and 88 of them were encoded by pseudogenes. In addition, bacterial infection of HL-60 cells up-regulated the expression of human proteins involved mostly in cytoskeleton components, vesicular trafficking, cell signaling, and energy metabolism, but down regulated some pattern recognition receptors involved in innate immunity. Our proteomics data represent a comprehensive analysis of A. phagocytophilum and E. chaffeensis proteomes, and provide a quantitative view of human host protein expression profiles regulated by bacterial infection. The availability of these proteomic data will provide new insights into biology and pathogenesis of these obligatory intracellular pathogens.

  19. Remodeling of tick cytoskeleton in response to infection with Anaplasma phagocytophilum.

    PubMed

    Cabezas-Cruz, Alejandro; Alberdi, Pilar; Valdes, James J; Villar, Margarita; de la Fuente, Jose

    2017-06-01

    The obligate intracellular pathogen Anaplasma phagocytophilum infects vertebrate and tick hosts. In this study, a genome-wide search for cytoskeleton components was performed in the tick vector, Ixodes scapularis. The available transcriptomics and proteomics data was then used to characterize the mRNA and protein levels of I. scapularis cytoskeleton components in response to A. phagocytophilum infection. The results showed that cytoskeleton components described in other model organisms were present in the I. scapularis genome. One type of intermediate filaments (lamin), a family of septins that was recently implicated in the cellular response to intracellular pathogens, and several members of motor proteins (kinesins and dyneins) that could be implicated in the cytoplasmic movements of A. phagocytophilum were found. The results showed that levels of tubulin, actin, septin, actin-related proteins and motor proteins were affected by A. phagocytophilum, probably to facilitate infection in I. scapularis. Functional studies demonstrated a role for selected cytoskeleton components in pathogen infection. These results provided a more comprehensive view of the cytoskeletal components involved in the response to A. phagocytophilum infection in ticks.

  20. Reptile infection with Anaplasma phagocytophilum, the causative agent of granulocytic anaplasmosis.

    PubMed

    Nieto, Nathan C; Foley, Janet E; Bettaso, Jamie; Lane, Robert S

    2009-10-01

    Granulocytic anaplasmosis (GA) is a potentially fatal tick-borne rickettsial disease that occurs sporadically in the far western United States. We evaluated the prevalence of Anaplasma phagocytophilum in multiple species of lizards and snakes from enzootic sites in northern California, described the infestation prevalence of its tick vector Ixodes pacificus on reptiles, and conducted an experimental challenge of western fence lizards (Sceloporus occidentalis) and Pacific gopher snakes (Pituophis catenifer) with A. phagocytophilum delivered via needle inoculation or tick bite. Both serologically and polymerase-chain reaction (PCR)-positive lizards (seroprevalence = 10.8%, PCR prevalence = 10.2%) and snakes (seroprevalence = 5.8%, PCR prevalence = 11.7%) were detected among wild-caught animals. A DNA sequence of the A. phagocytophilum groESL gene from a PCR-positive snake was 100% homologous to that of the human-derived A. phagocytophilum. Experimental attempts to infect naïve animals were unsuccessful for snakes (n = 2), but 1 of 12 lizards became infected for 1 wk only by tick bite. Xenodiagnostic I. pacificus larvae that fed on a PCR-positive lizard did not acquire or transmit rickettsiae. Our findings suggest that lizards and snakes are exposed to A. phagocytophilum by infected ticks, but that they do not serve as primary reservoir hosts of this rickettsia.

  1. Anaplasma phagocytophilum Infection Subverts Carbohydrate Metabolic Pathways in the Tick Vector, Ixodes scapularis

    PubMed Central

    Cabezas-Cruz, Alejandro; Alberdi, Pilar; Valdés, James J.; Villar, Margarita; de la Fuente, José

    2017-01-01

    The obligate intracellular pathogen, Anaplasma phagocytophilum, is the causative agent of human, equine, and canine granulocytic anaplasmosis and tick-borne fever (TBF) in ruminants. A. phagocytophilum has become an emerging tick-borne pathogen in the United States, Europe, Africa, and Asia, with increasing numbers of infected people and animals every year. It has been recognized that intracellular pathogens manipulate host cell metabolic pathways to increase infection and transmission in both vertebrate and invertebrate hosts. However, our current knowledge on how A. phagocytophilum affect these processes in the tick vector, Ixodes scapularis is limited. In this study, a genome-wide search for components of major carbohydrate metabolic pathways was performed in I. scapularis ticks for which the genome was recently published. The enzymes involved in the seven major carbohydrate metabolic pathways glycolysis, gluconeogenesis, pentose phosphate, tricarboxylic acid cycle (TCA), glyceroneogenesis, and mitochondrial oxidative phosphorylation and β-oxidation were identified. Then, the available transcriptomics and proteomics data was used to characterize the mRNA and protein levels of I. scapularis major carbohydrate metabolic pathway components in response to A. phagocytophilum infection of tick tissues and cultured cells. The results showed that major carbohydrate metabolic pathways are conserved in ticks. A. phagocytophilum infection inhibits gluconeogenesis and mitochondrial metabolism, but increases the expression of glycolytic genes. A model was proposed to explain how A. phagocytophilum could simultaneously control tick cell glucose metabolism and cytoskeleton organization, which may be achieved in part by up-regulating and stabilizing hypoxia inducible factor 1 alpha in a hypoxia-independent manner. The present work provides a more comprehensive view of the major carbohydrate metabolic pathways involved in the response to A. phagocytophilum infection in ticks

  2. Demonstration of transplacental transmission of a human isolate of Anaplasma phagocytophilum in an experimentally infected sheep.

    PubMed

    Reppert, E; Galindo, R C; Breshears, M A; Kocan, K M; Blouin, E F; de la Fuente, J

    2013-11-01

    Anaplasma phagocytophilum, first identified as a pathogen of sheep in Europe, more recently has been recognized as an emerging tick-borne pathogen of humans in the U.S. and Europe. Transmission of A. phagocytophilum is reported to be by ticks, primarily of the genus Ixodes. While mechanical and transplacental transmission of the type genus organism, A. marginale, occur in addition to tick transmission, these modes of transmission have not been considered for A. phagocytophilum. Recently, we developed a sheep model for studying host-tick-pathogen interactions of the human NY-18 A. phagocytophilum isolate. Sheep were susceptible to infection with this human isolate and served as a source of infection for I. scapularis ticks, but they did not display clinical signs of disease, and the pathogen was not apparent in stained blood smears. In the course of these experiments, one sheep unexpectedly gave birth to a lamb 5 weeks after being experimentally infected by inoculation with the pathogen propagated in HL-60 cells. The lamb was depressed and not feeding and was subsequently euthanized 18 h after birth. Tissues were collected at necropsy for microscopic examination and PCR to confirm A. phagocytophilum infection. At necropsy, the stomach contained colostrum, the spleen was moderately enlarged and thickened with conspicuous lymphoid follicles, and mesenteric lymph nodes were mildly enlarged and contained moderate infiltrates of eosinophils and neutrophils. Blood, spleen, heart, skin and cervical and mesenteric lymph nodes tested positive for A. phagocytophilum by PCR, and sequence analysis confirmed that the lamb was infected with the NY-18 isolate. Transplacental transmission should therefore be considered as a means of A. phagocytophilum transmission and may likely contribute to the epidemiology of tick-borne fever in sheep and other mammals, including humans.

  3. Candidatus Neoehrlichia mikurensis and Anaplasma phagocytophilum in natural rodent and tick communities in Southern Hungary.

    PubMed

    Szekeres, Sándor; Claudia Coipan, Elena; Rigó, Krisztina; Majoros, Gábor; Jahfari, Setareh; Sprong, Hein; Földvári, Gábor

    2015-03-01

    The aim of this study was to investigate the natural cycle of the new human pathogenic bacteria Candidatus Neoehrlichia mikurensis and Anaplasma phagocytophilum in Southern Hungary. We collected rodents with live-traps (2010-2013) and questing ticks with flagging in 2012. Small mammals were euthanized, tissue samples were collected and all the ectoparasites were removed and stored in 70% alcohol. We found relatively low overall prevalence of tick infestation (8%). Samples were analysed for A. phagocytophilum and Candidatus N. mikurensis with multiplex quantitative real-time PCR targeting a part of major surface protein 2 (msp2) and the heat shock protein groEL genes, respectively. The overall prevalence in tissue samples was 6.6% (skin) and 5.1% (spleen) for A. phagocytophilum and 1.7% (skin) and 3.4% (spleen) for Candidatus N. mikurensis. Candidatus N. mikurensis was only detected in Apodemus flavicollis and Apodemus agrarius, while A. phagocytophilum was found in A. flavicollis, A. agrarius, Myodes glareolus, Microtus arvalis and Mus musculus samples. Prevalence of A. phagocytophilum in skin samples of A. flavicollis was significantly higher than prevalence of N. mikurensis (p<0.05). Among questing Ixodes ricinus ticks we found three (8.8%) individuals (female, male, nymph) infected with Candidatus N. mikurensis. Five (3.1%) questing ticks had A. phagocytophilum infection (one I. ricinus male, two Dermacentor reticulatus females and two Haemaphysalis concinna females). We found one I. ricinus nymph removed from a male A. flavicollis with A. phagocytophilum infection. Our study provides new data on the occurrence of these pathogens in rodent tissue samples, questing ticks and engorged ticks in Southern Hungary.

  4. The infection of reintroduced ruminants - Bison bonasus and Alces alces - with Anaplasma phagocytophilum in northern Poland.

    PubMed

    Karbowiak, Grzegorz; Víchová, Bronislava; Werszko, Joanna; Demiaszkiewicz, Aleksander W; Pyziel, Anna M; Sytykiewicz, Hubert; Szewczyk, Tomasz; Peťko, Branislav

    2015-12-01

    The north-eastern part of Poland is considered an area of high risk for infection with tick-borne diseases, including with human granulocytic ehrlichiosis (HGE) agents. The etiological agent of HGE is Anaplasma phagocytophilum. As the animal reservoir for A. phagocytophilum in the environment serve the species from Cervidae and Bovidae families. European bison (Bison bonasus) and elk (Alces alces) are the big ruminant species, reintroduced to the forests of Middle Europe after many decades of absence. In the foci of zoonotic diseases they are able to play a role as natural reservoir to pathogens, however, their status as protected animals means their study has been rare and fragmentary. The studies of B. bonasus were conducted in Białowieża Primeval Forest and A. alces in Biebrza National Park. PCR amplifications were performed using primers amplifing the end of the groES gene, the intergenic spacer and approximately two-thirds of the groEL gene in the first round, and primers that span a 395-bp region of the groEL gene were used in the second round. The positive results were obtained in B. bonasus and A. alces, the prevalence of infection was 66.7 and 20.0%, respectively. Randomly selected samples were sequenced, sequences were compared with GenBank entries using Blast N2.2.13 and determined as A. phagocytophilum. The results presented herein are the first record of the presence of Anaplasma phagocytophilum in A. alces, and at the same time confirm the previous observations regarding the infection of B. bonasus with A. phagocytophilum.

  5. Ecology of Anaplasma phagocytophilum infection in gray foxes (Urocyon cinereoargenteus) in northwestern California.

    PubMed

    Gabriel, Mourad W; Brown, Richard N; Foley, Janet E; Higley, J Mark; Botzler, Richard G

    2009-04-01

    Although granulocytic anaplasmosis, caused by infection of Anaplasma phagocytophilum, is an emerging human and domestic animal disease, the ecology and natural history of the parasite is not well understood. Gray foxes (Urocyon cinereoargenteus) are relatively common, occasionally peri-urban mesocarnivores whose geographic distribution overlaps the reported distribution of granulocytic anaplasmosis in humans and domestic animals in North America. We evaluated the potential of foxes as hosts and reservoirs of A. phagocytophilum in both urban and backcountry habitats of the Hoopa Valley Indian Reservation, Humboldt County, California, USA. We trapped 54 individual foxes and had 16 recaptures for a total of 70 fox samples between June 2003 and October 2004 in delineated urban and backcountry zones. We collected 296 adult and 145 nymphal ticks from the 70 captured foxes including 193 Ixodes pacificus, 149 Ixodes texanus, 98 Dermacentor variabilis, and one Dermacentor occidentalis. There were seasonal differences in tick intensities, with most I. pacificus adults occurring in winter and spring (P < 0.001), most I. texanus nymphs in spring (P = 0.03), and most D. variabilis adults in spring and summer (P = 0.01). Thirty-six (51%) of the 70 fox sera had antibodies against A. phagocytophilum, with a higher (P = 0.24) prevalence in backcountry foxes (16 of 23) than in urban-zone foxes (12 of 31). Six (9%) of 70 fox samples were polymerase chain reaction-positive for A. phagocytophilum. Twenty-eight (31%) of 90 domestic dogs sampled from vaccine clinics within the study area were seropositive for A. phagocytophilum. There was a significant difference in prevalence between dogs and backcountry foxes (70%), but no differences were found between dogs and urban foxes (39%). We propose that gray foxes are a good sentinel species for A. phagocytophilum infections in northwestern California.

  6. First report of Anaplasma phagocytophilum and Babesia microti in rodents in Finland.

    PubMed

    Kallio, Eva R; Begon, Michael; Birtles, Richard J; Bown, Kevin J; Koskela, Esa; Mappes, Tapio; Watts, Phillip C

    2014-06-01

    Tick-borne diseases pose an increasingly important public health problem in Europe. Rodents are the reservoir host for many tick-transmitted pathogens, including Anaplasma phagocytophilum and Babesia microti, which can cause human granulocytic anaplasmosis and babesiosis, respectively. To estimate the presence of these pathogens in rodents in Finland, we examined blood samples from 151 bank voles (Myodes glareolus) and demonstrate, for the first time, that A. phagocytophilum and B. microti commonly infect bank voles (in 22% and 40% of animals, respectively) in Finland. Sequence analysis of a fragment of 18S rRNA showed that the B. microti strain isolated was identical to the Munich strain, which is considered to be nonzoonotic. The A. phagocytophilum strain (based on a fragment of the msp4 gene) was identical to one found earlier in rodents in the United Kingdom that is transmitted by the tick Ixodes trianguliceps, all the life stages of which feed on small mammals. The infection probability of B. microti in the bank voles was the greater the older the individual was, and males were more often infected than females. A. phagocytophilum infection probability first increased and then decreased with the age of individual without any difference between sexes. While these pathogens presumably pose a limited zoonotic risk to humans in Finland, they might have important interactions with other rodent pathogens and therefore affect infection dynamics of, for example, zoonotic pathogens.

  7. Evaluation of squirrels (Rodentia: Sciuridae) as ecologically significant hosts for Anaplasma phagocytophilum in California.

    PubMed

    Nieto, Nathan C; Foley, Janet E

    2008-07-01

    Granulocytic anaplasmosis (GA), caused by Anaplasma phagocytophilum, is a potentially fatal, emerging rickettsial disease of humans, domestic animals, and wildlife. The purpose of this study was to determine whether sciurids from multiple areas of northern California were infested with ticks or exposed to or infected with A. phagocytophilum using polymerase chain reaction (PCR) and indirect-fluorescent antibody (IFA) serology. Sciurids of nine different tree- and ground-dwelling species were assessed: arboreal squirrels (western and eastern gray squirrels, Sciurus griseus and S. carolinensis, and Douglas squirrels, Tamiasciurus douglasii) but not northern flying squirrels (Glaucomys sabrinus) had greater evidence of exposure and current infection than did semiarboreal or ground dwelling sciurids (California ground squirrels, Spermophilus beecheyi, and chipmunks, Tamias spp.). Western gray squirrels had the most extensive exposure (70.7% seroprevalence and 12.1% PCR prevalence). Positive squirrels were identified in all regions where squirrels were collected. A logistic regression identified being a western gray squirrel (OR = 20.5, P = 2.95 X 10(-8)) and from the north coastal region of California (OR = 9.052, P = 1.41 X 10(-6)) as having the highest risk of exposure to A. phagocytophilum. Five of nine sciurid species had evidence of infestation with Ixodes pacificus or I. spinipalpis that could vector A. phagocytophilum. Extensive exposure from multiple areas suggests sciurids may be important in the maintenance of GA in California and indicates that studies of reservoir competence of these species are warranted.

  8. First Report of Anaplasma phagocytophilum and Babesia microti in Rodents in Finland

    PubMed Central

    Begon, Michael; Birtles, Richard J.; Bown, Kevin J.; Koskela, Esa; Mappes, Tapio; Watts, Phillip C.

    2014-01-01

    Abstract Tick-borne diseases pose an increasingly important public health problem in Europe. Rodents are the reservoir host for many tick-transmitted pathogens, including Anaplasma phagocytophilum and Babesia microti, which can cause human granulocytic anaplasmosis and babesiosis, respectively. To estimate the presence of these pathogens in rodents in Finland, we examined blood samples from 151 bank voles (Myodes glareolus) and demonstrate, for the first time, that A. phagocytophilum and B. microti commonly infect bank voles (in 22% and 40% of animals, respectively) in Finland. Sequence analysis of a fragment of 18S rRNA showed that the B. microti strain isolated was identical to the Munich strain, which is considered to be nonzoonotic. The A. phagocytophilum strain (based on a fragment of the msp4 gene) was identical to one found earlier in rodents in the United Kingdom that is transmitted by the tick Ixodes trianguliceps, all the life stages of which feed on small mammals. The infection probability of B. microti in the bank voles was the greater the older the individual was, and males were more often infected than females. A. phagocytophilum infection probability first increased and then decreased with the age of individual without any difference between sexes. While these pathogens presumably pose a limited zoonotic risk to humans in Finland, they might have important interactions with other rodent pathogens and therefore affect infection dynamics of, for example, zoonotic pathogens. PMID:24848684

  9. Use of imidacloprid-permethrin to prevent transmission of Anaplasma phagocytophilum from naturally infected Ixodes scapularis ticks to dogs.

    PubMed

    Blagburn, Byron L; Spencer, Jennifer A; Billeter, Sarah A; Drazenovich, Nicole L; Butler, Jamie M; Land, Tracey M; Dykstra, Christine C; Stafford, Kirby C; Pough, Margaret B; Levy, Steven A; Bledsoe, David L

    2004-01-01

    One group of eight beagles was treated with a combination of imidacloprid and permethrin 7 days before exposure to Ixodes scapularis ticks that were naturally infected with Anaplasma phagocytophilum. A second group of eight beagles was not treated and was also exposed to infected ticks. Seven of eight non-treated dogs--but none of the treated dogs--developed specific antibodies to A. phagocytophilum. Results of this study indicate that a combination of imidacloprid and permethrin can prevent transmission of A. phagocytophilum to dogs if administered before exposure to infected ticks.

  10. Molecular analyses of a potentially novel Anaplasma species closely related to Anaplasma phagocytophilum detected in sika deer (Cervus nippon yesoensis) in Japan.

    PubMed

    Ybañez, Adrian P; Matsumoto, Kotaro; Kishimoto, Toshio; Inokuma, Hisashi

    2012-05-25

    An Anaplasma species closely related to Anaplasma phagocytophilum detected in sika deer in Hokkaido, Japan was molecularly analyzed using 16S rRNA, citrate synthase (gltA), and heat-shock operon (groEL) gene sequences. Genome walking was performed to determine its complete gltA and groEL sequences (1233 bp and 1650 bp, respectively). Percent identities to the closest A. phagocytophilum sequences from the US and European strains were 98.6-98.8%, 76.5%, and 80.3-80.8% for 16S rRNA, gltA, and groEL genes, respectively. For deduced amino acid sequences, percent identities to the closest A. phagocytophilum sequences were 66.7% and 97.6% for gltA and groEL genes, respectively. Phylogenetic analyses revealed divergence from any known A. phagocytophilum strain. The lower identities and the divergent phylogenetic position of the Anaplasma sp. detected from sika deer in Japan with established A. phagocytophilum strains provide evidence of its potential novelty. Copyright © 2011 Elsevier B.V. All rights reserved.

  11. Specific molecular detection of Anaplasma sp. closely related to Anaplasma phagocytophilum in ixodid ticks and cattle in a pastureland in Hokkaido, Japan.

    PubMed

    Ybañez, Adrian P; Tagawa, Michihito; Matsumoto, Kotaro; Kishimoto, Toshio; Yokoyama, Naoaki; Inokuma, Hisashi

    2013-01-01

    Recent molecular analyses of the Anaplasma sp. closely related to Anaplasma phagocytophilum (previously believed to be A. phagocytophilum) in Japan have clarified its distinct phylogenetic position. PCR methods relying on 16S rRNA- and P44/MSP2-based primers designed to detect this species have low sensitivity and specificity. In this study, a highly sensitive and specific nested PCR method using newly designed primers based on heat-shock operon gene (groEL) was developed to detect this species. The method was later used in an epidemiological study testing DNA samples from 85 Ixodid ticks (collected by flagging) and 50 cattle from the same pastureland in Nakaosobetsu, Hokkaido, Japan. Results revealed prevalence rates of 2.4% (2 of 85) in ticks and 2% (1 of 50) in cattle. The present study also reported the first molecular detection of the Anaplasma sp. closely related to A. phagocytophilum in Japan in H. douglasii, and established a new reliable PCR method that detects this Anaplasma sp. closely related to A. phagocytophilum in Japan.

  12. The dusky-footed woodrat (Neotoma fuscipes) is susceptible to infection by Anaplasma phagocytophilum originating from woodrats, horses, and dogs.

    PubMed

    Nieto, Nathan C; Madigan, John E; Foley, Janet E

    2010-07-01

    The dusky-footed woodrat (Neotoma fuscipes) is the putative reservoir for Anaplasma phagocytophilum in the western United States based on high prevalence of A. phagocytophilum genetic material detected by polymerase chain reaction (PCR), high antibody prevalence, and infestation with the vector tick Ixodes pacificus. Following inoculation of nine wild-caught woodrats with woodrat-, canine-, or equine-origin A. phagocytophilum, all became PCR-positive and seroconverted. However, when PCR-positive woodrat blood was delivered as inoculum to three horses, the horses never became clinically ill, PCR-positive, or antibody-positive. Each horse was subsequently infected with either the equine- or chipmunk-origin A. phagocytophilum, resulting in clinical anaplasmosis. Our data show that woodrats are permissive to several strains of A. phagocytophilum, but strains originating from woodrats did not cause clinical disease in horses.

  13. Detection of Anaplasma phagocytophilum in red foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) from Brandenburg, Germany.

    PubMed

    Härtwig, Vera; von Loewenich, Friederike D; Schulze, Christoph; Straubinger, Reinhard K; Daugschies, Arwid; Dyachenko, Viktor

    2014-04-01

    Anaplasma phagocytophilum is an obligate intracellular and tick-transmitted bacterium, which causes granulocytic anaplasmosis in animals and humans. Although infection with A. phagocytophilum in domestic animals and vector ticks is documented, there is sparse information on the occurrence of A. phagocytophilum in wild animals. Red foxes (Vulpes vulpes) as well as raccoon dogs (Nyctereutes procyonoides) are wildlife species highly abundant in certain areas of Germany and represent a potential wildlife reservoir for zoonotic diseases. To obtain data about the occurrence of A. phagocytophilum in these animals, red fox and raccoon dog carcasses (hunted or found dead) were collected from January to September 2009 in the Federal State of Brandenburg, Germany. Lung tissue samples were subjected to DNA extraction and were examined for the presence of A. phagocytophilum DNA by means of real-time PCR. Anaplasma phagocytophilum was detected in 10 out of 122 (8.2%) lungs of red foxes and in 3 out of 13 (23%) lungs of raccoon dogs. To the best of our knowledge, A. phagocytophilum was detected for the first time in red foxes and raccoon dogs in Germany. Copyright © 2014 Elsevier GmbH. All rights reserved.

  14. Anaplasma phagocytophilum and Ehrlichia chaffeensis type IV secretion and Ank proteins

    PubMed Central

    RIKIHISA, YASUKO; LIN, MINGQUN

    2011-01-01

    Summary of recent advances The obligatory intracellular bacterial pathogens Anaplasma and Ehrlichia infect leukocytes by hijacking host-cell components and processes. The type IV secretion system is up-regulated during infection. Among type IV secretion candidate substrates, an ankyrin repeat protein of Anaplasma phagocytophilum, AnkA, is delivered into the host cytoplasm via a complex that includes VirD4. AnkA is highly tyrosine-phosphorylated and binds to the Abl interactor 1, SHP-1, and nuclear DNA fragments. Ehrlichia chaffeensis AnkA was recently reported to be translocated into host cell nucleus. The recent discovery of several ankyrin repeat proteins secreted via the type IV secretion system of different intracellular bacteria suggests that a common strategy evolved to subvert host-cell functions. PMID:20053580

  15. Experimental infection in lambs with a red deer (Cervus elaphus) isolate of Anaplasma phagocytophilum.

    PubMed

    Stuen, Snorre; Scharf, Wiebke; Schauer, Sonja; Freyburger, Felix; Bergström, Karin; von Loewenich, Friederike D

    2010-07-01

    Anaplasma phagocytophilum transmitted by Ixodes spp. ticks is the causative agent of tick-borne fever (TBF) in domestic ruminants. TBF is widespread along the coast of southern Norway and may cause a severe problem for the sheep industry. Red deer (Cervus elaphus) are important hosts for ticks and have been found to be infected naturally with A. phagocytophilum. However, it is unclear whether red deer could serve as reservoir hosts for A. phagocytophilum infections in sheep. We infected lambs experimentally with a red deer and a sheep isolate, respectively. The 497 base pairs of the partial 16S rRNA gene sequences of both isolates were 100% identical to GenBank accession number M73220; the 3.8 kilobases of the total ank gene sequences were 99% identical. Sixteen lambs were used, four lambs in each group. Two groups were inoculated with the red deer isolate on day 0, and then challenged on day 42 with the ovine or the red deer isolate, respectively. The third group was inoculated with the sheep isolate on day 0 and challenged with the red deer strain on day 42. Four lambs were used as uninfected controls. Blood samples for hematology, bacteriology, and serology were collected regularly for 12 wk. Presence of A. phagocytophilum in blood was determined using blood smears. Serologic response was measured by indirect immunofluorescence. Although animals inoculated with the ovine strain showed more severe clinical manifestations, lambs infected with the red deer isolate reacted with typical signs of TBF such as fever, bacteremia, and neutropenia. We conclude that A. phagocytophilum strains causing TBF in sheep might circulate in the red deer population in Norway.

  16. Anaplasma phagocytophilum Asp14 Is an Invasin That Interacts with Mammalian Host Cells via Its C Terminus To Facilitate Infection

    PubMed Central

    Kahlon, Amandeep; Ojogun, Nore; Ragland, Stephanie A.; Seidman, David; Troese, Matthew J.; Ottens, Andrew K.; Mastronunzio, Juliana E.; Truchan, Hilary K.; Walker, Naomi J.; Borjesson, Dori L.; Fikrig, Erol

    2013-01-01

    Anaplasma phagocytophilum, a member of the family Anaplasmataceae, is the tick-transmitted obligate intracellular bacterium that causes human granulocytic anaplasmosis. The life cycle of A. phagocytophilum is biphasic, transitioning between the noninfectious reticulate cell (RC) and infectious dense-cored (DC) forms. We analyzed the bacterium's DC surface proteome by selective biotinylation of surface proteins, NeutrAvidin affinity purification, and mass spectrometry. Transcriptional profiling of selected outer membrane protein candidates over the course of infection revealed that aph_0248 (designated asp14 [14-kDa A. phagocytophilum surface protein]) expression was upregulated the most during A. phagocytophilum cellular invasion. asp14 transcription was induced during transmission feeding of A. phagocytophilum-infected ticks on mice and was upregulated when the bacterium engaged its receptor, P-selectin glycoprotein ligand 1. Asp14 localized to the A. phagocytophilum surface and was expressed during in vivo infection. Treating DC organisms with Asp14 antiserum or preincubating mammalian host cells with glutathione S-transferase (GST)–Asp14 significantly inhibited infection of host cells. Moreover, preincubating host cells with GST-tagged forms of both Asp14 and outer membrane protein A, another A. phagocytophilum invasin, pronouncedly reduced infection relative to treatment with either protein alone. The Asp14 domain that is sufficient for cellular adherence and invasion lies within the C-terminal 12 to 24 amino acids and is conserved among other Anaplasma and Ehrlichia species. These results identify Asp14 as an A. phagocytophilum surface protein that is critical for infection, delineate its invasion domain, and demonstrate the potential of targeting Asp14 in concert with OmpA for protecting against infection by A. phagocytophilum and other Anaplasmataceae pathogens. PMID:23071137

  17. Human Exposure to Anaplasma phagocytophilum in Two Cities of Northwestern Morocco

    PubMed Central

    Elhamiani Khatat, Sarah; Sahibi, Hamid; Hing, Mony; Alaoui Moustain, Ismail; El Amri, Hamid; Benajiba, Mohammed; Kachani, Malika; Duchateau, Luc; Daminet, Sylvie

    2016-01-01

    Anaplasma phagocytophilum is an emerging tick-borne zoonosis with extensive increased interest. Epidemiological data are available in several regions of the USA, Europe and Asia in contrast to other parts of the world such as North Africa. Blood samples of 261 healthy individuals divided in two groups i.e., dog handlers and blood donors were analysed. Indirect immunofluorescent assay using a commercial kit was performed to detect specific A. phagocytophilum IgG. Two dilutions were used to assess the prevalence of seroreactive samples. Demographic variables were assessed as potential risk factors using exact logistic regression. Seropositivity rates reached 37% and 27% in dog handlers and 36% and 22% in blood donors. No statistically significant differences were found in the prevalence rates between the two groups. Analysis of risk factors such as gender, age groups, outdoor activities, self-reported previous exposure to ticks, or contact with domestic animals (dogs, cats, ruminants and horses) did not shown any significant difference. A. phagocytophilum exposure was common in both high-risk population and blood donors in Morocco. PMID:27532208

  18. Epidemiology and Genetic Diversity of Anaplasma phagocytophilum in the San Francisco Bay Area, California

    PubMed Central

    Nieto, Nathan C.; Salkeld, Daniel J.

    2016-01-01

    In California, the agent of human granulocytic anaplasmosis (HGA), Anaplasma phagocytophilum, is transmitted by western black-legged ticks (Ixodes pacificus). Cases of HGA are infrequent in California but do occur annually. We investigated nymphal and adult western black-legged tick populations in 20 recreational areas in California's San Francisco Bay Area (Marin, Napa, San Mateo, Santa Clara, Santa Cruz, and Sonoma counties). Overall, prevalence of A. phagocytophilum in adult ticks was 0.8% (11/1,465), and in nymphal ticks was 4.2% (24/568), though presence was patchy and prevalence varied locally. We detected significant sequence variation in our quantitative polymerase chain reaction (qPCR)–positive samples. This included four sequences that grouped within a clade that contains clinical human and veterinary isolates as well as four others that grouped with sequences from PCR-positive lizards from northern California. Tick populations in our study sites harbor genetically diverse strains of A. phagocytophilum, which may influence potential risk in the region. PMID:27139447

  19. Anaplasma phagocytophilum Inhibits Apoptosis and Promotes Cytoskeleton Rearrangement for Infection of Tick Cells

    PubMed Central

    Ayllón, Nieves; Villar, Margarita; Busby, Ann T.; Kocan, Katherine M.; Blouin, Edmour F.; Bonzón-Kulichenko, Elena; Galindo, Ruth C.; Mangold, Atilio J.; Alberdi, Pilar; Pérez de la Lastra, José M.; Vázquez, Jesús

    2013-01-01

    Anaplasma phagocytophilum causes human granulocytic anaplasmosis. Infection with this zoonotic pathogen affects gene expression in both the vertebrate host and the tick vector, Ixodes scapularis. Here, we identified new genes, including spectrin alpha chain or alpha-fodrin (CG8) and voltage-dependent anion-selective channel or mitochondrial porin (T2), that are involved in A. phagocytophilum infection/multiplication and the tick cell response to infection. The pathogen downregulated the expression of CG8 in tick salivary glands and T2 in both the gut and salivary glands to inhibit apoptosis as a mechanism to subvert host cell defenses and increase infection. In the gut, the tick response to infection through CG8 upregulation was used by the pathogen to increase infection due to the cytoskeleton rearrangement that is required for pathogen infection. These results increase our understanding of the role of tick genes during A. phagocytophilum infection and multiplication and demonstrate that the pathogen uses similar strategies to establish infection in both vertebrate and invertebrate hosts. PMID:23630955

  20. Molecular Characterization of Anaplasma phagocytophilum and Borrelia burgdorferi in Ixodes scapularis Ticks from Pennsylvania

    PubMed Central

    Courtney, Joshua W.; Dryden, Richard L.; Montgomery, Jill; Schneider, Bradley S.; Smith, Gary; Massung, Robert F.

    2003-01-01

    Ixodes scapularis ticks were collected in 2000 and 2001 from two areas in Pennsylvania and tested for the presence of Anaplasma phagocytophilum and Borrelia burgdorferi by PCR and DNA sequencing. Of the ticks collected from northwestern and southeastern Pennsylvania, 162 of 263 (61.6%) and 25 of 191 (13.1%), respectively, were found to be positive for B. burgdorferi. DNA sequencing showed >99% identity with B. burgdorferi strains B31 and JD1. PCR testing for A. phagocytophilum revealed that 5 of 263 (1.9%) from northwestern Pennsylvania and 76 of 191 (39.8%) from southeastern Pennsylvania were positive. DNA sequencing revealed two genotypes of A. phagocytophilum, the human granulocytic ehrlichiosis (HGE) agent and a variant (AP-Variant 1) that has not been associated with human infection. Although only the HGE agent was present in northwestern Pennsylvania, both genotypes were found in southeastern Pennsylvania. These data add to a growing body of evidence showing that AP-Variant 1 is the predominant agent in areas where both genotypes coexist. PMID:12682147

  1. Molecular Events Involved in Cellular Invasion by Ehrlichia chaffeensis and Anaplasma phagocytophilum

    PubMed Central

    Rikihisa, Yasuko

    2009-01-01

    Ehrlichia chaffeensis and Anaplasma phagocytophilum are obligatory intracellular bacteria that preferentially replicate inside leukocytes by utilizing biological compounds and processes of these primary host defensive cells. These bacteria incorporate cholesterol from the host for their survival. Upon interaction with host monocytes and granulocytes, respectively, these bacteria usurp the lipid raft domain containing GPI-anchored protein to induce a series of signaling events that result in internalization of the bacteria. Monocytes and neutrophils usually kill invading microorganisms by fusion of the phagosomes containing the bacteria with granules containing both antimicrobial peptides and lysosomal hydrolytic enzymes and/or through sequestering vital nutrients. However, E. chaffeensis and A. phagocytophilum alter vesicular traffic to create a unique intracellular membrane-bound compartment that allows their replication in seclusion from lysosomal killing. These bacteria are quite sensitive to reactive oxygen species (ROS), so in order to survive in host cells that are primary mediators of ROS-induced killing, they inhibit activation of NADPH oxidase and assembly of this enzyme in their inclusion compartments. Moreover, host phagocyte activation and differentiation, apoptosis, and IFN-γ signaling pathways are inhibited by these bacteria. Through reductive evolution, lipopolysaccharide and peptidoglycan that activate the innate immune response, have been eliminated from these gram-negative bacteria at the genomic level. Upon interaction with new host cells, bacterial genes encoding the type IV secretion apparatus and the two-component regulatory system are upregulated to sense and adapt to the host environment. Thus dynamic signal transduction events concurrently proceed both in the host cells and in the invading E. chaffeensis and A. phagocytophilum bacteria for successful establishment of intracellular infection. Several bacterial surface exposed proteins and

  2. Molecular detection of Anaplasma platys, Anaplasma phagocytophilum and Wolbachia sp. but not Ehrlichia canis in Croatian dogs.

    PubMed

    Huber, Doroteja; Reil, Irena; Duvnjak, Sanja; Jurković, Daria; Lukačević, Damir; Pilat, Miroslav; Beck, Ana; Mihaljević, Željko; Vojta, Lea; Polkinghorne, Adam; Beck, Relja

    2017-09-14

    The bacteria Anaplasma platys, Anaplasma phagocytophilum and Ehrlichia canis are tick-borne agents that cause canine vector-borne disease. The prevalence of these pathogens in South Eastern Europe is unknown with the exception of an isolated case of A. platys detected in a dog imported into Germany from Croatia. To gain a better insight into their presence and prevalence, PCR-based screening for these bacterial pathogens was performed on domesticated dogs from different regions of Croatia. Blood samples from 1080 apparently healthy dogs from coastal and continental parts of Croatia as well as tissue samples collected from 63 deceased dogs with a history of anaemia and thrombocytopenia were collected for molecular screening by an Anaplasmataceae-specific 16S rRNA conventional PCR. Positive samples were confirmed using a second Anaplasmataceae-specific PCR assay with the PCR product sequenced for the purpose of bacterial species identification. All sequenced isolates were georeferenced and a kernel intensity estimator was used to identify clusters of greater case intensity. 42/1080 (3.8%; CI 2.7-5.0) of the healthy dogs were PCR positive for bacteria in the Anaplasmataceae. Sequencing of the 16S rRNA gene amplified from these positive samples revealed the presence of A. platys in 2.5% (CI 1.6-3.4%, 27 dogs), A. phagocytophilum in 0.3% (CI 0-0.6%, 3 dogs) and a Wolbachia endosymbiont in 1.1% (CI 0.4-1.6%, 12 dogs) of dogs screened in this study. Necropsied dogs were free from infection. Notably, no evidence of E. canis infection was found in any animal. This survey represents a rare molecular study of Anaplasmataceae in dogs in South Eastern Europe, confirming the presence of A. platys and A. phagocytophilum but not E. canis. The absence of E. canis was surprising given it has been described in all other Mediterranean countries surveyed and raises questions over the regional vector capacity of the Rhipicephalus sanguineus tick.

  3. Spatial distribution of Anaplasma phagocytophilum and Hepatozoon canis in red foxes (Vulpes vulpes) in Hungary.

    PubMed

    Tolnai, Z; Sréter-Lancz, Z; Sréter, T

    2015-07-01

    In recent years, Ehrlichia canis and Hepatozoon canis transmitted by Rhipicephalus sanguineus were reported from Hungary. The aim of the present study was to reveal the spatial distribution pattern of pathogens transmitted by R. sanguineus in a sentinel species, red fox (Vulpes vulpes) in Hungary and to analyse the relationship of these patterns with landscape and climate by geographical information systems. Fox carcasses, representing 0.5% of the total fox population were randomly selected out of all the foxes of Hungary. The spleen samples of the animals were tested by real-time PCR for Anaplasma platys, Babesia vogeli, E. canis and H. canis infection. Positive results were confirmed by conventional PCR followed by sequencing. The prevalence of H. canis infection was 22.2% (95% CI=18.4-26.4%), and this parasite was detected in all areas including the mountain regions of Hungary. These findings indicate that other tick species or other transmission routes (oral and transplacental) might be in the background of the countrywide distribution of H. canis. Anaplasma platys was not found; nevertheless, the prevalence of Anaplasma phagocytophilum infection transmitted by Ixodes ricinus was 12.5% (95% CI=9.7-16.1%) in foxes. B. vogeli and E. canis infection was not detected. There was no correlation between environmental parameter values in the home range of foxes and A. phagocytophilum or H. canis infection, which is in line with that observed in the case of tick species infesting foxes in Hungary. The results of this study indicate that R. sanguineus, if present, might be rare in Hungary. Our baseline study can be used for future evaluation of the effect of climate change on the spreading and emergence of R. sanguineus transmitted pathogens in Hungary. Copyright © 2015 Elsevier GmbH. All rights reserved.

  4. Wild boar (Sus scrofa) - reservoir host of Toxoplasma gondii, Neospora caninum and Anaplasma phagocytophilum in Slovakia.

    PubMed

    Reiterová, Katarína; Špilovská, Silvia; Blaňarová, Lucia; Derdáková, Markéta; Čobádiová, Andrea; Hisira, Vladimír

    2016-03-01

    In Central Europe the wild boar population is permanently growing and consequently Cf foodborne infections. In this study serological and molecular detection of Toxoplasma gondii and Neospora caninum in wild boars was evaluated. Moreover, same samples were screened for the presence and genetic variability of tick-borne bacterium Anaplasma phagocytophilum. Blood samples collected from 113 wild boars from Southern Slovakia were examined for antibodies to T. gondii by indirect and to N. caninum by competitive ELISA. The presence of parasitic DNA in blood samples was determined by standard or real time PCR techniques. Antibodies against T. gondii and N. caninum were detected in 45 (39.8%) and 38 (33.6%) animals, respectively. Females were more frequently infected for both pathogens than males. The high seropositivity against both coccidia indicates a permanent occurrence of these pathogens in the studied locality. T. gondii DNA was confirmed in five seropositive boars (4.4%) and N. caninum in 23 blood samples (20.4%). Three out of 23 N. caninum PCR positive animals did not show seropositivity. Three out of 113 blood samples of wild boars were positive for A. phagocytophilum (2.7%). The obtained A. phagocytophilum sequences were 100% identical with GenBankTM isolates from Slovak dog (KC985242); German horse (JF893938) or wild boar (EF143810) and red deer (EF143808) from Poland. Coinfections of T. gondii with N. caninum and N. caninum with A. phagocytophilum were detected in single cases. Results suggest a potential zoonotic risk of toxoplasmosis transmission to humans and the spread of neosporosis to farm animals.

  5. First molecular detection and phylogenetic analysis of Anaplasma phagocytophilum in shelter dogs in Seoul, Korea.

    PubMed

    Lee, Sukyee; Lee, Seung-Hun; VanBik, Dorene; Kim, Neung-Hee; Kim, Kyoo-Tae; Goo, Youn-Kyoung; Rhee, Man Hee; Kwon, Oh-Deog; Kwak, Dongmi

    2016-07-01

    In this study, the status of Anaplasma phagocytophilum infection was assessed in shelter dogs in Seoul, Korea, with PCR and phylogenetic analyses. Nested PCR on 1058 collected blood samples revealed only one A. phagocytophilum positive sample (female, age <1year, mixed breed, collected from the north of the Han River). The genetic variability of A. phagocytophilum was evaluated by genotyping, using the 16S rRNA, groEL, and msp2 gene sequences of the positive sample. BLASTn analysis revealed that the 16S rRNA, groEL, and msp2 genes had 99.6%, 99.9%, and 100% identity with the following sequences deposited in GenBank: a cat 16S rRNA sequence from Korea (KR021166), a rat groEL sequence from Korea (KT220194), and a water deer msp2 sequence from Korea (HM752099), respectively. Phylogenetic analyses classified the groEL gene into two distinct groups (serine and alanine), whereas the msp2 gene showed a general classification into two groups (USA and Europe) that were further subgrouped according to region. To the best of our knowledge, this study is the first to describe the molecular diagnosis of A. phagocytophilum in dogs reared in Korea. In addition, the high genetic identity of the 16S rRNA and groEL sequences between humans and dogs from the same region suggests a possible epidemiological relation. Given the conditions of climate change, tick ecology, and recent incidence of human granulocytic anaplasmosis in Korea, the findings of this study underscore the need to establish appropriate control programs for tick-borne diseases in Korea.

  6. The European hedgehog (Erinaceus europaeus)--a suitable reservoir for variants of Anaplasma phagocytophilum?

    PubMed

    Silaghi, Cornelia; Skuballa, Jasmin; Thiel, Claudia; Pfister, Kurt; Petney, Trevor; Pfäffle, Miriam; Taraschewski, Horst; Passos, Lygia M F

    2012-02-01

    The European hedgehog (Erinaceus europaeus) is a common insectivore in most parts of Europe and is frequently infested by the ticks Ixodes ricinus and I. hexagonus. I. ricinus ticks have been found infected with Anaplasma phagocytophilum, an obligate intracellular bacterium, but little is known about the potential of the hedgehog as a reservoir host. In this study, the infection with A. phagocytophilum and the genetic variants involved were investigated in a captive hedgehog population which was kept in a fenced, natural grass and bush garden habitat, and also in its ticks. Additionally hedgehogs from hedgehog caretaking stations were investigated. EDTA blood and ticks were collected from the captive hedgehog population once a month from March to October 2007 and in March and April 2008. All 3 developmental stages of I. ricinus and I. hexagonus occurred on the hedgehogs. After DNA extraction, the samples were screened for A. phagocytophilum with a real-time PCR, and selected samples were further investigated with a nested PCR targeting the partial 16S rRNA gene, followed by sequencing. One hundred thirty-six out of 220 hedgehog blood samples (61.8%) from altogether 48 individuals, 413 out of 563 I. ricinus samples and 90 out of 338 I. hexagonus samples were PCR-positive. Thirty-two hedgehogs were positive more than once, most frequently twice or 3 times, but also up to 9 times. Sequencing of the partial 16S rRNA gene resulted in 6 variants, but one variant ('A') was the most frequent which appeared in 93.8% of the positive hedgehogs. This variant (equaling Frankonia II, GenBank AF136712) has recently been reported from human, equine, and canine granulocytic anaplasmosis cases and thus, its specific association with hedgehogs is an important finding in the epidemiology of A. phagocytophilum in Europe. The high infection rate of both hedgehogs and ticks with A. phagocytophilum and the simultaneous infestation with 2 tick species of all developmental stages suggest

  7. Sheep experimentally infected with a human isolate of Anaplasma phagocytophilum serve as a host for infection of Ixodes scapularis ticks.

    PubMed

    Kocan, Katherine M; Busby, Ann T; Allison, Robin W; Breshears, Melanie A; Coburn, Lisa; Galindo, Ruth C; Ayllón, Nieves; Blouin, Edmour F; de la Fuente, José

    2012-06-01

    Anaplasma phagocytophilum, first identified as a pathogen of ruminants in Europe, has more recently been recognized as an emerging tick-borne pathogen of humans in the U.S. and Europe. A. phagocytophilum is transmitted by Ixodes spp., but the tick developmental cycle and pathogen/vector interactions have not been fully described. In this research, we report on the experimental infection of sheep with the human NY-18 isolate of A. phagocytophilum which then served as a host for infection of I. scapularis nymphs and adults. A. phagocytophilum was propagated in the human promyelocytic cell line, HL-60, and the infected cell cultures were then used to infect sheep by intravenous inoculation. Infections in sheep were confirmed by PCR and an Anaplasma-competitive ELISA. Clinical signs were not apparent in any of the infected sheep, and only limited hematologic and mild serum biochemical abnormalities were identified. While A. phagocytophilum morulae were rarely seen in neutrophils, blood film evaluation revealed prominent large granular lymphocytes, occasional plasma cells, and rare macrophages. Upon necropsy, gross lesions were restricted to the lymphoid system. Mild splenomegaly and lymphadenomegaly with microscopic evidence of lymphoid hyperplasia was observed in all infected sheep. Female I. scapularis that were allowed to feed and acquire infection on each of the 3 experimentally infected sheep became infected with A. phagocytophilum as determined by PCR of guts (80-87%) and salivary glands (67-100%). Female I. scapularis that acquired infection as nymphs on an experimentally infected sheep transmitted A. phagocytophilum to a susceptible sheep, thus confirming transstadial transmission. Sheep proved to be a good host for the production of I. scapularis infected with this human isolate of A. phagocytophilum, which can be used as a model for future studies of the tick/pathogen interface.

  8. The first report of Anaplasma phagocytophilum and a novel Theileria spp. co-infection in a South African giraffe.

    PubMed

    Zhang, Yan; Li, Tongyi; Cui, Yanyan; Wang, Jinhong; Lv, Yali; Wang, Rongjun; Jian, Fuchun; Zhang, Longxian; Wang, Jiantang; Yang, Guangcheng; Ning, Changshen

    2016-08-01

    Organisms of the genera Anaplasma and Theileria are important intracellular bacteria and parasites that cause various tick-borne diseases, threatening the health of numerous animals as well as human beings. In the present study, a 12-month-old male wild South African giraffe (Giraffa camelopardalis giraffa) originating from South Africa, and living in Zhengzhou Zoo (located in the urban district of Zhengzhou in the provincial capital of Henan), suddenly developed an unknown fatal disease and died 1day after the onset of the clinical signs. By microscopic examination of Giemsa-stained blood smears combined with nested PCR and DNA sequence analysis, Anaplasma phagocytophilum, Anaplasma bovis and a novel Theileria spp. were found in the blood of this giraffe. The six other Cervidae animals in the zoo and three ruminants living in the same colony house with them were found to be negative for both Anaplasma and Theileria in their blood specimens. We report on the first case of an A. phagocytophilum infection and the occurrence of a novel Theileria spp. in the blood of a giraffe. This is the first reported case of a multi-infection of A. bovis, A. phagocytophilum and Theileria spp. in a giraffe, as revealed by microscopic examination of blood smears and the results of nested PCR and DNA sequencing.

  9. [Lyme borreliosis and co-infections. Place of Anaplasma phagocytophilum and Bartonella henselae].

    PubMed

    Christmann, Daniel

    2015-01-01

    Lyme borreliosis (LB) is certainly the most common infection transmitted through the bite of Ixodes in Northern Hemisphere. These ticks are also able to transmit other microorganisms such as the bacteria Anaplasma phagocytophilum (Ap) and Bartonella henselae (Bh), with the latter discovered fairly recently, leading to diferent clinical presentations often close to those of LB. The aim of this study was to evaluate the frequency of co-infection by either of these bacteria in patients with LB, particularly when a treatment with beta-lactam antibiotic was only partially effective. Of these patients, on the basis of serological data, 8.07% were simultaneously contaminated by Bh, 6.83% by Ap and 4.96% were co-infected by Bh and Ap. Since the choice of an antibiotic should take into account the specificities of these germs and especially their intracellular proliferation, these results should be considered in selecting treatment.

  10. The Tick Salivary Protein Sialostatin L2 Inhibits Caspase-1-Mediated Inflammation during Anaplasma phagocytophilum Infection

    PubMed Central

    Chen, Gang; Wang, Xiaowei; Severo, Maiara S.; Sakhon, Olivia S.; Sohail, Mohammad; Brown, Lindsey J.; Sircar, Mayukh; Snyder, Greg A.; Sundberg, Eric J.; Ulland, Tyler K.; Olivier, Alicia K.; Andersen, John F.; Zhou, Yi; Shi, Guo-Ping; Sutterwala, Fayyaz S.; Kotsyfakis, Michail

    2014-01-01

    Saliva from arthropod vectors facilitates blood feeding by altering host inflammation. Whether arthropod saliva counters inflammasome signaling, a protein scaffold that regulates the activity of caspase-1 and cleavage of interleukin-1β (IL-1β) and IL-18 into mature molecules, remains elusive. In this study, we provide evidence that a tick salivary protein, sialostatin L2, inhibits inflammasome formation during pathogen infection. We show that sialostatin L2 targets caspase-1 activity during host stimulation with the rickettsial agent Anaplasma phagocytophilum. A. phagocytophilum causes macrophage activation and hemophagocytic syndrome features. The effect of sialostatin L2 in macrophages was not due to direct caspase-1 enzymatic inhibition, and it did not rely on nuclear factor κB or cathepsin L signaling. Reactive oxygen species from NADPH oxidase and the Loop2 domain of sialostatin L2 were important for the regulatory process. Altogether, our data expand the knowledge of immunoregulatory pathways of tick salivary proteins and unveil an important finding in inflammasome biology. PMID:24686067

  11. The first clinical and laboratory evidence of co-infection by Anaplasma phagocytophilum and Ehrlichia canis in a Brazilian dog.

    PubMed

    Silveira, Júlia A G; Valente, Pâmela C L G; Paes, Paulo R O; Vasconcelos, Artur V; Silvestre, Bruna T; Ribeiro, Múcio F B

    2015-04-01

    Information on Anaplasma phagocytophilum in Brazil is very restricted. The aim of this study was to report clinical, parasitological, hematological and molecular evidence of a natural A. phagocytophilum infection of an urban Brazilian dog. The dog was an eight-month-old male French bulldog. Veterinary clinical examinations were performed three times: in April, June and December 2013. Biochemical and hematological analyses were performed during all examinations, and blood samples were collected for parasitological surveys in June and December. Morulae were present within neutrophils in blood smears from June. Both samples were PCR positive for A. phagocytophilum and Ehrlichia spp. Phylogenetic analysis revealed that the phylogenetic topology placed samples from this study in close proximity to other A. phagocytophilum isolates. Ehrlichia isolates from this dog were 100% identical to E. canis isolates, thus E. canis and A. phagocytophilum co-infection was diagnosed in this dog. Lethargy and skin lesions were the clinical signs observed in this dog. Abnormal hematological parameters, among those, severe thrombocytopenia, were observed in all three occasions. This finding highlights the growing importance of A. phagocytophilum in South America.

  12. The Prostaglandin E2-EP3 Receptor Axis Regulates Anaplasma phagocytophilum-Mediated NLRC4 Inflammasome Activation

    PubMed Central

    Wang, Xiaowei; Shaw, Dana K.; Hammond, Holly L.; Sutterwala, Fayyaz S.; Rayamajhi, Manira; Shirey, Kari Ann; Perkins, Darren J.; Bonventre, Joseph V.; Velayutham, Thangam S.; Evans, Sean M.; Rodino, Kyle G.; VieBrock, Lauren; Scanlon, Karen M.; Carbonetti, Nicholas H.; Carlyon, Jason A.; Miao, Edward A.; McBride, Jere W.; Kotsyfakis, Michail

    2016-01-01

    Rickettsial agents are sensed by pattern recognition receptors but lack pathogen-associated molecular patterns commonly observed in facultative intracellular bacteria. Due to these molecular features, the order Rickettsiales can be used to uncover broader principles of bacterial immunity. Here, we used the bacterium Anaplasma phagocytophilum, the agent of human granulocytic anaplasmosis, to reveal a novel microbial surveillance system. Mechanistically, we discovered that upon A. phagocytophilum infection, cytosolic phospholipase A2 cleaves arachidonic acid from phospholipids, which is converted to the eicosanoid prostaglandin E2 (PGE2) via cyclooxygenase 2 (COX2) and the membrane associated prostaglandin E synthase-1 (mPGES-1). PGE2-EP3 receptor signaling leads to activation of the NLRC4 inflammasome and secretion of interleukin (IL)-1β and IL-18. Importantly, the receptor-interacting serine/threonine-protein kinase 2 (RIPK2) was identified as a major regulator of the immune response against A. phagocytophilum. Accordingly, mice lacking COX2 were more susceptible to A. phagocytophilum, had a defect in IL-18 secretion and exhibited splenomegaly and damage to the splenic architecture. Remarkably, Salmonella-induced NLRC4 inflammasome activation was not affected by either chemical inhibition or genetic ablation of genes associated with PGE2 biosynthesis and signaling. This divergence in immune circuitry was due to reduced levels of the PGE2-EP3 receptor during Salmonella infection when compared to A. phagocytophilum. Collectively, we reveal the existence of a functionally distinct NLRC4 inflammasome illustrated by the rickettsial agent A. phagocytophilum. PMID:27482714

  13. Evidence of Anaplasma phagocytophilum and Borrelia burgdorferi infection in cats after exposure to wild-caught adult Ixodes scapularis.

    PubMed

    Lappin, Michael R; Chandrashekar, Ramaswamy; Stillman, Brett; Liu, Jiayou; Mather, Thomas N

    2015-07-01

    Cats are infected by Anaplasma phagocytophilum and Borrelia burgdorferi when exposed to infected Ixodes scapularis (black-legged ticks). The purpose of our study was to allow wild-caught I. scapularis to feed on healthy research cats (n = 4) and temporally evaluate for A. phagocytophilum DNA in blood by a polymerase chain reaction (PCR) assay as well as for antibody responses to the B. burgdorferi C6 peptide, to the A. phagocytophilum P44 peptide, and to a novel A. phagocytophilum peptide (P44-4). Prior to I. scapularis infestation, all cats were negative for antibodies against both organisms based on a kit optimized for dog serum, and negative for A. phagocytophilum DNA in blood using a conventional PCR assay. Using the pre-infestation samples, an enzyme-linked immunosorbent assay for detecting antibodies against the P44-4 peptide was optimized. Cats were infested with wild-caught I. scapularis for 7 days. Genomic DNA of A. phagocytophilum was amplified from the blood before antibodies were detected in all 4 cats. Antibodies against the C6 peptide, P44 peptide, and P44-4 peptide were detected in the sera of all 4 cats. Antibodies against P44-4 were detected prior to those against P44 in 3 out of 4 cats. The results suggest that a PCR assay should be considered in acutely ill cats with suspected anaplasmosis that are seronegative.

  14. In vitro cultivation of Anaplasma marginale and A. phagocytophilum in tick cell lines: a review.

    PubMed

    Passos, Lygia Maria Friche

    2012-01-01

    Continuous cell lines have been established from several ixodid and argasid tick species, representing an excellent tool suitable for the isolation of pathogens and their subsequent propagation, which in turn allows the production of antigenic material for diagnostic tests, antibody and vaccine production, and also for studies on host-vector-pathogen relationships. This paper reviews the use of tick cells for culture initiation and maintenance of two obligate intracellular bacterial pathogens, Anaplasma marginale and Anaplasma phagocytophilum. These in vitro cultivation systems have been used in a wide range of studies, covering morphological ultrastructural analysis, genetics, proteomics and biological differences between strains, including genome transcriptional and protein expression approaches, enabling comparisons between host and vector cells. Thus, such systems open a new window for a better understanding of interactions between pathogens and tick cells. Last but not least, such systems contribute to the reduction in usage of animals for experimental research, as antigenic material can be produced in reasonably large quantities without the use of in vivo species-specific systems.

  15. The infection of questing Dermacentor reticulatus ticks with Babesia canis and Anaplasma phagocytophilum in the Chernobyl exclusion zone.

    PubMed

    Karbowiak, Grzegorz; Vichová, Bronislavá; Slivinska, Kateryna; Werszko, Joanna; Didyk, Julia; Peťko, Branislav; Stanko, Michal; Akimov, Igor

    2014-08-29

    Tick occurrence was studied in the Chernobyl exclusion zone (CEZ) during the August-October 2009-2012. Dermacentor reticulatus ticks were collected using the flagging method and then screened for infection with Anaplasma phagocytophilum and Babesia canis by a PCR method incorporating specific primers and sequence analysis. The prevalence of infection with B. canis canis and A. phagocytophilum was found to be 3.41% and 25.36%, respectively. The results present the first evidence of B. canis canis and A. phagocytophilum in questing D. reticulatus ticks from the Chernobyl exclusion zone. They also reveal the presence of tick-borne disease foci in areas with no human activity, and confirm that they can be maintained in areas after a nuclear disaster with radioactive contamination.

  16. Anaplasma phagocytophilum induces Ixodes scapularis ticks to express an antifreeze glycoprotein gene that enhances their survival in the cold

    PubMed Central

    Neelakanta, Girish; Sultana, Hameeda; Fish, Durland; Anderson, John F.; Fikrig, Erol

    2010-01-01

    In the United States, Ixodes scapularis ticks overwinter in the Northeast and Upper Midwest and transmit the agent of human granulocytic anaplasmosis, Anaplasma phagocytophilum, among other pathogens. We now show that the presence of A. phagocytophilum in I. scapularis ticks increases their ability to survive in the cold. We identified an I. scapularis antifreeze glycoprotein, designated IAFGP, and demonstrated via RNAi knockdown studies the importance of IAFGP for the survival of I. scapularis ticks in a cold environment. Transfection studies also show that IAFGP increased the viability of yeast cells subjected to cold temperature. Remarkably, A. phagocytophilum induced the expression of iafgp, thereby increasing the cold tolerance and survival of I. scapularis. These data define a molecular basis for symbiosis between a human pathogenic bacterium and its arthropod vector and delineate what we believe to be a new pathway that may be targeted to alter the life cycle of this microbe and its invertebrate host. PMID:20739755

  17. Seroprevalence of antibodies to tick-borne encephalitis virus and Anaplasma phagocytophilum in healthy adults from western Norway.

    PubMed

    Hjetland, Reidar; Henningsson, Anna J; Vainio, Kirsti; Dudman, Susanne G; Grude, Nils; Ulvestad, Elling

    2015-01-01

    The aim of this study was to assess the seroprevalence of antibodies to tick-borne encephalitis virus (TBEV) and Anaplasma phagocytophilum in a healthy adult population from Sogn and Fjordane county in western Norway. Sera from 1, 213 blood donors were analysed for IgG-antibodies to TBEV, and a random subgroup of 301 donors for IgG to A. phagocytophilum. In the TBEV ELISA, five (0.4%) sera were positive. These were all interpreted as "false" positives, as four had received vaccines against flaviviruses, and the remaining was negative for neutralizing antibodies to TBEV. Antibodies to A. phagocytophilum were detected by indirect immunofluorescence in 49 (16.2%) subjects (titer range 80-1280). The results indicate that TBE currently is not endemic in this part of western Norway. However, there is serological evidence of the existence of human granulocytic anaplasmosis in the population.

  18. Anaplasma phagocytophilum infection in American robins and gray catbirds: an assessment of reservoir competence and disease in captive wildlife.

    PubMed

    Johnston, Emily; Tsao, Jean I; Muñoz, Juan David; Owen, Jen

    2013-01-01

    Anaplasma phagocytophilum (Dumler et al.) is the bacterial agent of human granulocytic anaplasmosis, an emerging infectious disease. The main vector of A. phagocytophilum in the United States is the blacklegged tick (Ixodes scapularis (Say)) and various small and medium-sized mammals are reservoirs. Previous studies indicate that birds are exposed to A. phagocytophilum; however, because no studies have directly investigated avian susceptibility, reservoir competence, and morbidity for A. phagocytophilum, uncertainty remains as to what role birds could play in its transmission ecology. In a controlled laboratory study, we tested whether two species, the American robin (Turdus migratorius (L.)) and the gray catbird (Dumetella carolinensis (L.)), can become infected with and then transmit A. phagocytophilum to feeding ticks, and whether exposed birds develop disease. Wild caught, seronegative birds (n = 10 per species) were exposed to A. phagocytophilum-infected I. scapularis nymphs (day 0). Transmission was assessed by xenodiagnosis on days 7, 14, 42, and 77; blood was assayed for bacteremia and serology. A. phagocytophilum was detected using quantitative polymerase chain reaction targeting the 16s rRNA gene. One robin infected 2 of 13 larval ticks (15%) on day 7; no other birds were found to infect feeding ticks at any time. Birds did not develop bacteremia, specific antibodies or significant illness because of exposure. Mouse controls became bacteremic, infected feeding ticks, and seroconverted. Our results suggest that these two avian species are unlikely to play a significant role in the maintenance of the agent of human granulocytic anaplasmosis and that avian serosurveys may not be a reliable indicator of A. phagocytophilum exposure.

  19. Nidicolous ticks of small mammals in Anaplasma phagocytophilum-enzootic sites in northern California

    PubMed Central

    Foley, Janet; Rejmanek, Daniel; Fleer, Katryna; Nieto, Nathan

    2011-01-01

    Ixodes spp. tick-borne zoonotic diseases are present across the Holarctic in humans, domestic animals, and wildlife. Small mammals are reservoirs for the rickettsial pathogen Anaplasma phagocytophilum and tick vectors may include catholic-feeding bridge vectors as well as host-specialist or nidicolous ticks. Far western North American communities in which A. phagocytophilum is maintained are complex ecologically, with multiple reservoir host and tick species, multiple strains of the bacterial pathogen A. phagocytophilum and differences in dynamics of hosts and vectors across heterogeneous landscapes. We evaluated sites in northern California in order to identify primarily nidicolous ticks and the hosts they infest. A total of 667 ticks was found in 11 study sites, including 288 on flags and 379 attached to small mammals. Larvae were over-represented among attached ticks and adults on flags. The most abundant species was I. pacificus. Two-hundred fourteen nidicolous ticks were found, most abundantly I. angustus and I. spinipalpis. All adult I. ochotonae, I. auritulus, I. angustus, I. jellisoni, and I. woodi were female, while the male:female ratio of I. spinipalpis was 1.2:1 and 1:1 for I. pacificus. The greatest number of ticks was obtained from Tamias ochrogenys, Peromyscus spp., and Neotoma fuscipes. Of 234 small mammal individuals that were infested with Ixodes spp., only 81 (34.6%) were infested with I. pacificus. The remaining infested small mammals hosted nidicolous tick species. Eight ticks were PCR-positive, including 6 I. pacificus (one adult, one larva, and 6 nymphs), and 2 adult I. ochotonae and high PCR prevalences of 18% and 9% were detected in woodrats and chipmunks, respectively. Nymphal I. angustus ticks were active year-long with a possible increase in August while larval activity was only observed in December and spring months and adults only during spring and fall. Overall, we show high tick species richness and year-round high levels of

  20. Molecular epidemiology of the emerging zoonosis agent Anaplasma phagocytophilum (Foggie, 1949) in dogs and ixodid ticks in Brazil

    PubMed Central

    2013-01-01

    Background Anaplasma phagocytophilum is an emerging pathogen of humans, dogs and other animals, and it is transmitted by ixodid ticks. The objective of the current study was a) detect A. phagocytophilum in dogs and ixodid ticks using real-time Polymerase Chain Reaction (qPCR); and b) Determine important variables associated to host, environment and potential tick vectors that are related to the presence of A. phagocytophilum in dogs domiciled in Rio de Janeiro, Brazil. Methods We tested blood samples from 398 dogs and samples from 235 ticks, including 194 Rhipicephalus sanguineus sensu lato, 15 Amblyomma cajennense, 8 Amblyomma ovale and 18 pools of Amblyomma sp. nymphs. A semi-structured questionnaire was applied by interviewing each dog owner. Deoxyribonucleic acid obtained from ticks and dog buffy coat samples were amplified by qPCR (msp2 gene). The sequencing of 16S rRNA and groESL heat shock operon genes and a phylogenetic analysis was performed. The multiple logistic regression model was created as a function of testing positive dogs for A. phagocytophilum. Results Among the 398 blood samples from dogs, 6.03% were positive for A. phagocytophilum. Anaplasma phagocytophilum was detected in one A. cajennense female tick and in five R. sanguineus sensu lato ticks (four males and one female). The partial sequences of the 16S rRNA, and groESL genes obtained were highly similar to strains of A. phagocytophilum isolated from wild birds from Brazil and human pathogenic strains. The tick species collected in positive dogs were R. sanguineus sensu lato and A. cajennense, with A.cajennense being predominant. Tick infestation history (OR = 2.86, CI = 1.98-14.87), dog size (OR = 2.41, IC: 1.51-12.67), the access to forest areas (OR = 3:51, CI: 1.52-16.32), hygiene conditions of the environment in which the dogs lived (OR = 4.35, CI: 1.86-18.63) and Amblyomma sp. infestation (OR = 6.12; CI: 2.11-28.15) were associated with A. phagocytophilum

  1. Molecular epidemiology of the emerging zoonosis agent Anaplasma phagocytophilum (Foggie, 1949) in dogs and ixodid ticks in Brazil.

    PubMed

    Santos, Huarrisson A; Thomé, Sandra M G; Baldani, Cristiane D; Silva, Claudia B; Peixoto, Maristela P; Pires, Marcus S; Vitari, Gabriela L V; Costa, Renata L; Santos, Tiago M; Angelo, Isabele C; Santos, Leandro A; Faccini, João L H; Massard, Carlos L

    2013-12-11

    Anaplasma phagocytophilum is an emerging pathogen of humans, dogs and other animals, and it is transmitted by ixodid ticks. The objective of the current study was a) detect A. phagocytophilum in dogs and ixodid ticks using real-time Polymerase Chain Reaction (qPCR); and b) Determine important variables associated to host, environment and potential tick vectors that are related to the presence of A. phagocytophilum in dogs domiciled in Rio de Janeiro, Brazil. We tested blood samples from 398 dogs and samples from 235 ticks, including 194 Rhipicephalus sanguineus sensu lato, 15 Amblyomma cajennense, 8 Amblyomma ovale and 18 pools of Amblyomma sp. nymphs. A semi-structured questionnaire was applied by interviewing each dog owner. Deoxyribonucleic acid obtained from ticks and dog buffy coat samples were amplified by qPCR (msp2 gene). The sequencing of 16S rRNA and groESL heat shock operon genes and a phylogenetic analysis was performed. The multiple logistic regression model was created as a function of testing positive dogs for A. phagocytophilum. Among the 398 blood samples from dogs, 6.03% were positive for A. phagocytophilum. Anaplasma phagocytophilum was detected in one A. cajennense female tick and in five R. sanguineus sensu lato ticks (four males and one female). The partial sequences of the 16S rRNA, and groESL genes obtained were highly similar to strains of A. phagocytophilum isolated from wild birds from Brazil and human pathogenic strains. The tick species collected in positive dogs were R. sanguineus sensu lato and A. cajennense, with A.cajennense being predominant. Tick infestation history (OR = 2.86, CI = 1.98-14.87), dog size (OR = 2.41, IC: 1.51-12.67), the access to forest areas (OR = 3:51, CI: 1.52-16.32), hygiene conditions of the environment in which the dogs lived (OR = 4.35, CI: 1.86-18.63) and Amblyomma sp. infestation (OR = 6.12; CI: 2.11-28.15) were associated with A. phagocytophilum infection in dogs. This is the

  2. Transmission route efficacy and kinetics of Anaplasma phagocytophilum infection in white-footed mouse, Peromyscus leucopus.

    PubMed

    Massung, Robert F; Priestley, Rachael A; Levin, Michael L

    2004-01-01

    Anaplasma phagocytophilum was used to infect Peromyscus leucopus mice by three routes of inoculation: infected tick infestation and intraperitoneal (IP) and subcutaneous (SQ) injection of infected tissue culture cells. A set of 12 mice were infected (four tick, four IP, and four SQ), and blood was drawn at 1, 3, 6, 9, 12, 15, 21, 28, 35, and 60 days post-infection and analyzed by use of a quantitative PCR assay to assess the level of infection. An additional set of 108 mice were infected (36 tick, 36 IP, 36 SQ) and euthanized at 1, 3, 6, 9, 12, 15, 21, 28, and 35 days post-infection (four mice/time point), and blood, spleen, bone marrow, and bladder tissue samples were analyzed. Tick infection generally produced the highest average levels of infection and peaked at 9 days post-infestation in blood, spleen, and bone marrow and at 6 days after infestation in the bladder. IP injection resulted in levels of infection that peaked on day 6 (spleen) or 12 (bladder, bone marrow, and blood). A. phagocytophilum injected SQ showed low levels of infection, and the day of peak infection varied. The average level of infection in the blood drawstressed mice was consistently higher and peaked earlier than infection in the non-stressed, euthanized mice. Xenodiagnosis was used to assay a third set of 12 mice (four tick, four IP, and four SQ) on days 7 and 14 post-infection and ticks fed on tick-infected mice showed the highest rate of PCR-positive test results at both time points (day 7, 22.2%; day 14, 17.3%). These data indicate that P. leucopus mice can be infected by tick infestation, IP injection, or SQ injection but that the kinetics and level of infection are quite variable among individual mice, may be influenced by the route of inoculation, and may be further altered by common laboratory procedures such as repeated collection of blood samples.

  3. An O-Methyltransferase Is Required for Infection of Tick Cells by Anaplasma phagocytophilum

    PubMed Central

    Oliva Chávez, Adela S.; Fairman, James W.; Felsheim, Roderick F.; Nelson, Curtis M.; Herron, Michael J.; Higgins, LeeAnn; Burkhardt, Nicole Y.; Oliver, Jonathan D.; Markowski, Todd W.; Kurtti, Timothy J.; Edwards, Thomas E.; Munderloh, Ulrike G.

    2015-01-01

    Anaplasma phagocytophilum, the causative agent of Human Granulocytic Anaplasmosis (HGA), is an obligately intracellular α-proteobacterium that is transmitted by Ixodes spp ticks. However, the pathogen is not transovarially transmitted between tick generations and therefore needs to survive in both a mammalian host and the arthropod vector to complete its life cycle. To adapt to different environments, pathogens rely on differential gene expression as well as the modification of proteins and other molecules. Random transposon mutagenesis of A. phagocytophilum resulted in an insertion within the coding region of an o-methyltransferase (omt) family 3 gene. In wild-type bacteria, expression of omt was up-regulated during binding to tick cells (ISE6) at 2 hr post-inoculation, but nearly absent by 4 hr p.i. Gene disruption reduced bacterial binding to ISE6 cells, and the mutant bacteria that were able to enter the cells were arrested in their replication and development. Analyses of the proteomes of wild-type versus mutant bacteria during binding to ISE6 cells identified Major Surface Protein 4 (Msp4), but also hypothetical protein APH_0406, as the most differentially methylated. Importantly, two glutamic acid residues (the targets of the OMT) were methyl-modified in wild-type Msp4, whereas a single asparagine (not a target of the OMT) was methylated in APH_0406. In vitro methylation assays demonstrated that recombinant OMT specifically methylated Msp4. Towards a greater understanding of the overall structure and catalytic activity of the OMT, we solved the apo (PDB_ID:4OA8), the S-adenosine homocystein-bound (PDB_ID:4OA5), the SAH-Mn2+ bound (PDB_ID:4PCA), and SAM- Mn2+ bound (PDB_ID:4PCL) X-ray crystal structures of the enzyme. Here, we characterized a mutation in A. phagocytophilum that affected the ability of the bacteria to productively infect cells from its natural vector. Nevertheless, due to the lack of complementation, we cannot rule out secondary mutations

  4. An O-Methyltransferase Is Required for Infection of Tick Cells by Anaplasma phagocytophilum.

    PubMed

    Oliva Chávez, Adela S; Fairman, James W; Felsheim, Roderick F; Nelson, Curtis M; Herron, Michael J; Higgins, LeeAnn; Burkhardt, Nicole Y; Oliver, Jonathan D; Markowski, Todd W; Kurtti, Timothy J; Edwards, Thomas E; Munderloh, Ulrike G

    2015-01-01

    Anaplasma phagocytophilum, the causative agent of Human Granulocytic Anaplasmosis (HGA), is an obligately intracellular α-proteobacterium that is transmitted by Ixodes spp ticks. However, the pathogen is not transovarially transmitted between tick generations and therefore needs to survive in both a mammalian host and the arthropod vector to complete its life cycle. To adapt to different environments, pathogens rely on differential gene expression as well as the modification of proteins and other molecules. Random transposon mutagenesis of A. phagocytophilum resulted in an insertion within the coding region of an o-methyltransferase (omt) family 3 gene. In wild-type bacteria, expression of omt was up-regulated during binding to tick cells (ISE6) at 2 hr post-inoculation, but nearly absent by 4 hr p.i. Gene disruption reduced bacterial binding to ISE6 cells, and the mutant bacteria that were able to enter the cells were arrested in their replication and development. Analyses of the proteomes of wild-type versus mutant bacteria during binding to ISE6 cells identified Major Surface Protein 4 (Msp4), but also hypothetical protein APH_0406, as the most differentially methylated. Importantly, two glutamic acid residues (the targets of the OMT) were methyl-modified in wild-type Msp4, whereas a single asparagine (not a target of the OMT) was methylated in APH_0406. In vitro methylation assays demonstrated that recombinant OMT specifically methylated Msp4. Towards a greater understanding of the overall structure and catalytic activity of the OMT, we solved the apo (PDB_ID:4OA8), the S-adenosine homocystein-bound (PDB_ID:4OA5), the SAH-Mn2+ bound (PDB_ID:4PCA), and SAM- Mn2+ bound (PDB_ID:4PCL) X-ray crystal structures of the enzyme. Here, we characterized a mutation in A. phagocytophilum that affected the ability of the bacteria to productively infect cells from its natural vector. Nevertheless, due to the lack of complementation, we cannot rule out secondary mutations.

  5. Prevalence and Diversity among Anaplasma phagocytophilum Strains Originating from Ixodes ricinus Ticks from Northwest Norway

    PubMed Central

    Tveten, Ann-Kristin

    2014-01-01

    The tick-borne pathogen Anaplasma phagocytophilum causes great concern for livestock farmers. Tick-borne fever is a widespread disease in Norway, and antibodies have been produced amongst sheep, roe deer, red deer, and moose. The main vector Ixodes ricinus is found along the Norwegian coastline as far north as the Arctic Circle. A total number of 1804 I. ricinus ticks were collected and the prevalence of the pathogen was determined by species-specific qPCR. The overall infection rate varied from 2.83% to 3.32%, but there were no significant differences (p = 0.01) in the overall infection rate in 2010, 2011, or 2012. A multilocus sequencing analysis was performed to further characterise the isolates. The genotyping of 27 strains resulted in classification into 19 different sequences types (ST), none of which was found in the MLST database. The nucleotide diversity was for every locus <0.01, and the number of SNPs was between 1 and 2.8 per 100 bp. The majority of SNPs were synonymous. A goeBURST analysis demonstrated that the strains from northwest Norway cluster together with other Norwegian strains in the MLST database and the strains that are included in this study constitute clonal complexes (CC) 9, 10, and 11 in addition to the singleton. PMID:25215241

  6. Multilocus sequence analysis of Anaplasma phagocytophilum reveals three distinct lineages with different host ranges in clinically ill French cattle.

    PubMed

    Chastagner, Amélie; Dugat, Thibaud; Vourc'h, Gwenaël; Verheyden, Hélène; Legrand, Loïc; Bachy, Véronique; Chabanne, Luc; Joncour, Guy; Maillard, Renaud; Boulouis, Henri-Jean; Haddad, Nadia; Bailly, Xavier; Leblond, Agnès

    2014-12-09

    Molecular epidemiology represents a powerful approach to elucidate the complex epidemiological cycles of multi-host pathogens, such as Anaplasma phagocytophilum. A. phagocytophilum is a tick-borne bacterium that affects a wide range of wild and domesticated animals. Here, we characterized its genetic diversity in populations of French cattle; we then compared the observed genotypes with those found in horses, dogs, and roe deer to determine whether genotypes of A. phagocytophilum are shared among different hosts. We sampled 120 domesticated animals (104 cattle, 13 horses, and 3 dogs) and 40 wild animals (roe deer) and used multilocus sequence analysis on nine loci (ankA, msp4, groESL, typA, pled, gyrA, recG, polA, and an intergenic region) to characterize the genotypes of A. phagocytophilum present. Phylogenic analysis revealed three genetic clusters of bacterial variants in domesticated animals. The two principal clusters included 98% of the bacterial genotypes found in cattle, which were only distantly related to those in roe deer. One cluster comprised only cattle genotypes, while the second contained genotypes from cattle, horses, and dogs. The third contained all roe deer genotypes and three cattle genotypes. Geographical factors could not explain this clustering pattern. These results suggest that roe deer do not contribute to the spread of A. phagocytophilum in cattle in France. Further studies should explore if these different clusters are associated with differing disease severity in domesticated hosts. Additionally, it remains to be seen if the three clusters of A. phagocytophilum genotypes in cattle correspond to distinct epidemiological cycles, potentially involving different reservoir hosts.

  7. Systems Biology of Tissue-Specific Response to Anaplasma phagocytophilum Reveals Differentiated Apoptosis in the Tick Vector Ixodes scapularis

    PubMed Central

    Ayllón, Nieves; Villar, Margarita; Galindo, Ruth C.; Kocan, Katherine M.; Šíma, Radek; López, Juan A.; Vázquez, Jesús; Alberdi, Pilar; Cabezas-Cruz, Alejandro; Kopáček, Petr; de la Fuente, José

    2015-01-01

    Anaplasma phagocytophilum is an emerging pathogen that causes human granulocytic anaplasmosis. Infection with this zoonotic pathogen affects cell function in both vertebrate host and the tick vector, Ixodes scapularis. Global tissue-specific response and apoptosis signaling pathways were characterized in I. scapularis nymphs and adult female midguts and salivary glands infected with A. phagocytophilum using a systems biology approach combining transcriptomics and proteomics. Apoptosis was selected for pathway-focused analysis due to its role in bacterial infection of tick cells. The results showed tissue-specific differences in tick response to infection and revealed differentiated regulation of apoptosis pathways. The impact of bacterial infection was more pronounced in tick nymphs and midguts than in salivary glands, probably reflecting bacterial developmental cycle. All apoptosis pathways described in other organisms were identified in I. scapularis, except for the absence of the Perforin ortholog. Functional characterization using RNA interference showed that Porin knockdown significantly increases tick colonization by A. phagocytophilum. Infection with A. phagocytophilum produced complex tissue-specific alterations in transcript and protein levels. In tick nymphs, the results suggested a possible effect of bacterial infection on the inhibition of tick immune response. In tick midguts, the results suggested that A. phagocytophilum infection inhibited cell apoptosis to facilitate and establish infection through up-regulation of the JAK/STAT pathway. Bacterial infection inhibited the intrinsic apoptosis pathway in tick salivary glands by down-regulating Porin expression that resulted in the inhibition of Cytochrome c release as the anti-apoptotic mechanism to facilitate bacterial infection. However, tick salivary glands may promote apoptosis to limit bacterial infection through induction of the extrinsic apoptosis pathway. These dynamic changes in response to A

  8. Anaplasma phagocytophilum MSP4 and HSP70 Proteins Are Involved in Interactions with Host Cells during Pathogen Infection

    PubMed Central

    Contreras, Marinela; Alberdi, Pilar; Mateos-Hernández, Lourdes; Fernández de Mera, Isabel G.; García-Pérez, Ana L.; Vancová, Marie; Villar, Margarita; Ayllón, Nieves; Cabezas-Cruz, Alejandro; Valdés, James J.; Stuen, Snorre; Gortazar, Christian; de la Fuente, José

    2017-01-01

    Anaplasma phagocytophilum transmembrane and surface proteins play a role during infection and multiplication in host neutrophils and tick vector cells. Recently, A. phagocytophilum Major surface protein 4 (MSP4) and Heat shock protein 70 (HSP70) were shown to be localized on the bacterial membrane, with a possible role during pathogen infection in ticks. In this study, we hypothesized that A. phagocytophilum MSP4 and HSP70 have similar functions in tick-pathogen and host-pathogen interactions. To address this hypothesis, herein we characterized the role of these bacterial proteins in interaction and infection of vertebrate host cells. The results showed that A. phagocytophilum MSP4 and HSP70 are involved in host-pathogen interactions, with a role for HSP70 during pathogen infection. The analysis of the potential protective capacity of MSP4 and MSP4-HSP70 antigens in immunized sheep showed that MSP4-HSP70 was only partially protective against pathogen infection. This limited protection may be associated with several factors, including the recognition of non-protective epitopes by IgG in immunized lambs. Nevertheless, these antigens may be combined with other candidate protective antigens for the development of vaccines for the control of human and animal granulocytic anaplasmosis. Focusing on the characterization of host protective immune mechanisms and protein-protein interactions at the host-pathogen interface may lead to the discovery and design of new effective protective antigens. PMID:28725639

  9. A putative marker for human pathogenic strains of Anaplasma phagocytophilum correlates with geography and host, but not human tropism.

    PubMed

    Foley, Janet; Stephenson, Nicole; Cubilla, Michelle Pires; Qurollo, Barbara; Breitschwerdt, Edward B

    2016-03-01

    Anaplasma phagocytophilum is an Ixodes species tick-transmitted bacterium that is capable of infecting a variety of host species, although there is a diversity of bacterial strains with differing host tropism. Recent analysis of A. phagocytophilum strains suggested that "drhm", a gene locus designated "distantly related to human marker" (drhm), which was predicted to be an integral membrane protein with possible transporter functions was not present in available canine and human isolates. By assessing 117 strains from 14 host species from across the US, we extended this analysis. Phylogenetic clades were associated with geography, but not host species. Additionally, a virulent clade that lacks drhm and infects dogs, horses, and humans in northeastern US was identified.

  10. [Prevalence of haemotropic Mycoplasma spp., Bartonella spp. and Anaplasma phagocytophilum in cats in Berlin/Brandenburg (Northeast Germany)].

    PubMed

    Morgenthal, Dinah; Hamel, Dietmar; Arndt, Gisela; Silaghi, Cornelia; Pfister, Kurt; Kempf, Volkhard A J; Kohn, Barbara

    2012-01-01

    Aim of this study was to evaluate the occurrence of Mycoplasma (M.) haemofelis, Candidatus Mycoplasma (C. M.) turicensis, C M. haemominutum, Bartonella spp. (B. henselae, B. clarridgeiae and B. quintana) and Anaplasma (A.) phagocytophilum in cats in Northeast Germany in relation to their living conditions (indoor/outdoor/ stray cat), and tick/flea exposure. 265 cats were included in the study (150 indoor, 99 outdoor access, 16 stray cats). A questionnaire provided the following data: derivation, housing environment, and previous flea/tick exposure. Serum antibody titers against A. phagocytophilum, B. henselae, and B. quintana were determined by an immunofluorescence test (IFT). PCR tests (EDTA blood) were used to test for A. phagocytophilum, M. haemofelis, C. M. turicensis, C. M. haemominutum, B. henselae and B. clarridgeiae. In 19 of 265 cats (7.2%) DNA of one or more Mycoplasma spp. was detected: C M. haemominutum (5.3%), M. haemofelis (1.5%) and C M. turicensis (1.1%); three of the cats were tested positive for the feline immunodeficiency virus. All cats were B. henselae and B. clarridgeiae PCR-negative in peripheral blood. However, 91 of 245 cats (37.1%) had antibody titers > 1:200 for B. henselae (Houston I, Marseille type) and 46 (18.8%) for B. quintana. Antibody titers > 1:64 against A. phagocytophilum were detected in 24 cats (9.1%); one cat (0.4%) was PCR-positive. Since infections with haemotropic Mycoplasma spp. and also with arthropodborne organisms (Bartonella spp., A. phagocytophilum) occur in cats from the area Berlin/Brandenburg (Germany) an appropriate arthropod-control is recommended. Further studies are needed to evaluate the relevance of these infectious agents for the individual cat.

  11. Comparison of the reservoir competence of medium-sized mammals and Peromyscus leucopus for Anaplasma phagocytophilum in Connecticut.

    PubMed

    Levin, Michael L; Nicholson, William L; Massung, Robert F; Sumner, John W; Fish, Durland

    2002-01-01

    In the northeastern United States, Anaplasma phagocytophilum, the agent of human granulocytic ehrlichiosis (HGE), is transmitted by the tick vector Ixodes scapularis. The white-footed mouse Peromyscus leucopus is a competent reservoir for this agent, but the reservoir competence of non-Peromyscus hosts of I. scapularis has not been studied. Here, we report data confirming reservoir competence of medium-sized mammals for A. phagocytophilum. Raccoons, Virginia opossums, gray squirrels, and striped skunks were live-trapped in June-August of 1998-1999 at two locations in Connecticut. Captured animals were kept for several days at the laboratory in wire-mesh cages over water to allow naturally attached ticks to drop off. Samples of blood and serum were taken from each animal prior to its release at the site of capture. Engorged ticks collected from each animal were allowed to molt. Resulting I. scapularis nymphs and adults were tested for the presence of A. phagocytophilum DNA by polymerase chain reaction, as were the blood samples from the animals. A. phagocytophilum DNA was detected in the blood of >10% of the raccoons tested. Raccoons, opossums, squirrels, and skunks produced adult I. scapularis infected with the agent of HGE. Prevalence of infection was the highest in adult ticks fed as nymphs upon raccoons (23%) and the lowest in those fed upon skunks and opossums (5-7%). The agent was present in nymphal I. scapularis fed as larvae upon raccoons and squirrels, but not in ticks fed upon skunks or opossums. We also tested the ability of I. scapularis to transmit A. phagocytophilum to laboratory-reared white-footed mice after acquiring it from medium-sized mammals. Ticks that acquired the agent from raccoons and squirrels successfully transmitted it to mice. Thus, raccoons and gray squirrels are reservoir-competent for the agent of HGE-they become naturally infected, and are capable of transmitting the infection to feeding ticks.

  12. [Monoinfections caused by Borrelia burgdorferi and Borrelia burgdorferi / Anaplasma phagocytophilum co-infections in forestry workers and farmers].

    PubMed

    Tokarska-Rodak, Małgorzata; Pańczuk, Anna; Kozioł-Montewka, Maria; Plewik, Dorota; Szepeluk, Adam

    2015-01-01

    The presence of co-infections induced by tick-borne pathogens in humans is an important epidemiological phenomenon. This issue has attracted growing attention of doctors and people working under conditions of an increased risk of being exposed to tick bites. The research group consisted of 93 individuals with current anti-immunoglobulin M/G (IgM/ IgG) Borrelia burgdorferi or IgG anti-Anaplasma phagocytophilum. The respondents were identified during the screening survey in a group of farmers and foresters occupationally exposed to tick bites. The aim of the work was to analyse the frequency of antibodies to specific antigens of B. burgdorferi and the levels of cytokines in forestry workers and farmers with B. burgdorferi monoinfections and B. burgdorferi / A. phagocytophilum co-infections. Statistical analysis was performed using the Chi2, Mann-Whitney U and Kruskal-Wallis tests. There is a stronger generation of IgG antibodies to B. burgdorferi antigens in patients with B. burgdorferi / A. phagocytophilum co-infections, such as variable major protein-like sequence expressed (VlsE) (p < 0.05), p19 (p < 0.02), p17 (p < 0.05) and complement regulator-acquiring surface protein 3 (CRASP3) (p < 0.02) compared to persons with B. burgdorferi monoinfections. The discrepancies in the synthesis of cytokines interleukin 6 (IL-6), IL-10, and tumor necrosis factor α (TNF-α) have not been found in persons with B. burgdorferi monoinfections and B. burgdorferi / A. phagocytophilum co-infection. The immune response directed against B. burgdorferi is stronger in patients co-infected with B. burgdorferi and A. phagocytophilum than in those with monoinfection. This work is available in Open Access model and licensed under a CC BY-NC 3.0 PL license.

  13. Molecular detection of Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato genospecies in red foxes (Vulpes vulpes) from Romania.

    PubMed

    Dumitrache, Mirabela Oana; Matei, Ioana Adriana; Ionică, Angela Monica; Kalmár, Zsuzsa; D'Amico, Gianluca; Sikó-Barabási, Sándor; Ionescu, Dan Traian; Gherman, Călin Mircea; Mihalca, Andrei Daniel

    2015-10-08

    Red foxes (Vulpes vulpes) are one of the most widespread wild carnivores in the world, being recognized to harbor and transmit a wide range of vector-borne diseases. Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato are zoonotic tick-borne pathogens causing emerging diseases. Wild animals play an essential role in the transmission of diseases and pathogens maintenance in nature. Epidemiological studies regarding the prevalence of tick-borne pathogens in red foxes are of public health importance, as they may successfully act as a pathogen transmission interface between wildlife, domestic animals and humans. This study included 14 counties from Romania. A total number of 353 red foxes (Vulpes vulpes) were examined. Heart tissue samples were collected during necropsy and stored at -20 °C. Genomic DNA extraction was performed and all samples were examined by polymerase chain reaction (PCR). Specific primers for A. phagocytophilum, A. platys, E. canis and Borrelia burgdorferi s.l. were used. Sequence analysis was performed (Macrogen Europe, Amsterdam) and obtained sequences are available at GenBank™. Out of the 353 samples, 9 (2.55 %; 95 % CI: 1.25-4.96 %) were positive for A. phagocytophilum. Positive animals originated from 5 counties. In total, 5 out of 353 heart tissue samples (1.42 %; 95 % CI: 0.52-3.47 %) collected from red foxes were positive for B. burgdorferi s.l. Red foxes originated from 4 counties. None of the samples were positive for A. platys or E. canis. No co-infection with A. phagocytophilum and B. burgdorferi s.l. was found. This first report of A. phagocytophilum and B. burgdorferi s.l. in red foxes from Romania suggests a limited role of foxes in the maintenance of the two related pathogens, but may represent a potential risk from a public health perspective.

  14. Transmission of Anaplasma phagocytophilum from endothelial cells to peripheral granulocytes in vitro under shear flow conditions

    PubMed Central

    Wang, Jinyong; Dyachenko, Viktor; Munderloh, Ulrike G.

    2015-01-01

    Anaplasma phagocytophilum(Ap) is a tick-borne pathogen, which can cause granulocytic anaplasmosis in humans and animals. In vivo this obligate intracellular pathogen is primarily located in circulating mature granulocytes, but it also infects endothelial cells. In order to study the interaction between Ap-infected endothelial cells and human granulocytes under conditions similar to those found naturally in the infected host, an in vitro model that mimics physiological flow conditions in the microvasculature was established. Cell-to-cell interactions were then visualized by microscopy, which showed that granulocytes adhered strongly to Ap-infected endothelial cells at a shear stress of 0.5 dyne/cm2. In addition, Ap-transmission assays under flow conditions showed that the bacteria transferred from infected endothelial cells to circulating granulocytes and were able to establish infection in constantly moving granulocytes. Cell surface analysis showed that Ap induced up-regulation of the cell adhesion molecules ICAM-1 and VCAM-1 on infected endothelial cells in a dose-dependent manner. Furthermore, IL-8 secretion by endothelial cells indicated that the presence of Ap induced a pro-inflammatory response. In summary, the results of this study suggest that endothelial cells of the microvasculature (1) provide an excellent site for Ap dissemination to peripheral blood granulocytes under flow conditions and therefore may play a crucial role in the development of persistent infection, and (2) are stimulated by Ap to express surface molecules and cytokines that may lead to inflammatory responses at the site of the infection. PMID:25618174

  15. Vaccinomics Approach to the Identification of Candidate Protective Antigens for the Control of Tick Vector Infestations and Anaplasma phagocytophilum Infection.

    PubMed

    Contreras, Marinela; Alberdi, Pilar; Fernández De Mera, Isabel G; Krull, Christoph; Nijhof, Ard; Villar, Margarita; De La Fuente, José

    2017-01-01

    Anaplasma phagocytophilum is an emerging tick-borne pathogen causing human granulocytic anaplasmosis (HGA), tick-borne fever (TBF) in small ruminants, and other forms of anaplasmosis in different domestic and wild animals. The main vectors of this pathogen are Ixodes tick species, particularly I. scapularis in the United States and I. ricinus in Europe. One of the main limitations for the development of effective vaccines for the prevention and control of A. phagocytophilum infection and transmission is the identification of effective tick protective antigens. The objective of this study was to apply a vaccinomics approach to I. scapularis-A. phagocytophilum interactions for the identification and characterization of candidate tick protective antigens for the control of vector infestations and A. phagocytophilum infection. The vaccinomics pipeline included the use of quantitative transcriptomics and proteomics data from uninfected and A. phagocytophilum-infected I. scapularis ticks for the selection of candidate protective antigens based on the variation in tick mRNA and protein levels in response to infection, their putative biological function, and the effect of antibodies against these proteins on tick cell apoptosis and pathogen infection. The characterization of selected candidate tick protective antigens included the identification and characterization of I. ricinus homologs, functional characterization by different methodologies including RNA interference, immunofluorescence, gene expression profiling, and artificial tick feeding on rabbit antibodies against the recombinant antigens to select the candidates for vaccination trials. The vaccinomics pipeline developed in this study resulted in the identification of two candidate tick protective antigens that could be selected for future vaccination trials. The results showed that I. scapularis lipocalin (ISCW005600) and lectin pathway inhibitor (AAY66632) and I. ricinus homologs constitute candidate protective

  16. Vaccinomics Approach to the Identification of Candidate Protective Antigens for the Control of Tick Vector Infestations and Anaplasma phagocytophilum Infection

    PubMed Central

    Contreras, Marinela; Alberdi, Pilar; Fernández De Mera, Isabel G.; Krull, Christoph; Nijhof, Ard; Villar, Margarita; De La Fuente, José

    2017-01-01

    Anaplasma phagocytophilum is an emerging tick-borne pathogen causing human granulocytic anaplasmosis (HGA), tick-borne fever (TBF) in small ruminants, and other forms of anaplasmosis in different domestic and wild animals. The main vectors of this pathogen are Ixodes tick species, particularly I. scapularis in the United States and I. ricinus in Europe. One of the main limitations for the development of effective vaccines for the prevention and control of A. phagocytophilum infection and transmission is the identification of effective tick protective antigens. The objective of this study was to apply a vaccinomics approach to I. scapularis-A. phagocytophilum interactions for the identification and characterization of candidate tick protective antigens for the control of vector infestations and A. phagocytophilum infection. The vaccinomics pipeline included the use of quantitative transcriptomics and proteomics data from uninfected and A. phagocytophilum-infected I. scapularis ticks for the selection of candidate protective antigens based on the variation in tick mRNA and protein levels in response to infection, their putative biological function, and the effect of antibodies against these proteins on tick cell apoptosis and pathogen infection. The characterization of selected candidate tick protective antigens included the identification and characterization of I. ricinus homologs, functional characterization by different methodologies including RNA interference, immunofluorescence, gene expression profiling, and artificial tick feeding on rabbit antibodies against the recombinant antigens to select the candidates for vaccination trials. The vaccinomics pipeline developed in this study resulted in the identification of two candidate tick protective antigens that could be selected for future vaccination trials. The results showed that I. scapularis lipocalin (ISCW005600) and lectin pathway inhibitor (AAY66632) and I. ricinus homologs constitute candidate protective

  17. Evaluation of Antibiotic Susceptibilities of Ehrlichia canis, Ehrlichia chaffeensis, and Anaplasma phagocytophilum by Real-Time PCR

    PubMed Central

    Branger, S.; Rolain, J. M.; Raoult, D.

    2004-01-01

    We determined MICs of antibiotics against Anaplasma phagocytophilum, Ehrlichia chaffeensis, and Ehrlichia canis by real-time quantitative PCR. The doubling times of the organisms were established: 19 h for E. chaffeensis, 26 h for A. phagocytophilum, and 28 h for E. canis. In comparison to the reference method for determining sensitivities, which uses Diff-Quick staining, our PCR assay was very sensitive and specific. We confirmed that doxycycline and rifampin are highly active against these bacteria and found variable susceptibilities to fluoroquinolones; A. phagocytophilum was susceptible, but E. canis and E. chaffeensis were only partly susceptible. β-Lactam compounds, cotrimoxazole, macrolide compounds, and telithromycin showed no activity against any of the three organisms. Thiamphenicol was found to be more active than chloramphenicol. For the first time, we showed that these three species have numerous point mutations in their 23S RNA genes, with those at positions 754, 2057, 2058, 2059, and 2611 (Escherichia coli numbering) known to confer resistance to macrolide compounds in other bacteria. The role of each of these mutations in resistance to these drugs should be investigated in the future. Our study confirms previous reports that quantitative PCR is a reliable method for determining antibiotic susceptibility; therefore, it might be useful for screening new drugs. PMID:15561862

  18. Candidatus Neoehrlichia mikurensis and Anaplasma phagocytophilum: prevalences and investigations on a new transmission path in small mammals and ixodid ticks.

    PubMed

    Obiegala, Anna; Pfeffer, Martin; Pfister, Kurt; Tiedemann, Tim; Thiel, Claudia; Balling, Anneliese; Karnath, Carolin; Woll, Dietlinde; Silaghi, Cornelia

    2014-12-04

    Small mammals are crucial for the life history of ixodid ticks, but their role and importance in the transmission cycle of tick-borne pathogens is mostly unknown. Candidatus Neoehrlichia mikurensis (CNM) and Anaplasma phagocytophilum are both tick-borne pathogens, and rodents are discussed to serve as main reservoir hosts for CNM but not for the latter especially in Germany. Analysing the prevalence of both pathogens in small mammals and their ticks in endemic regions may help to elucidate possible transmission paths in small mammal populations and between small mammals and ticks. In 2012 and 2013, small mammals were trapped at three different sites in Germany. DNA was extracted from different small mammal tissues, from rodent neonates, foetuses and from questing and attached ticks. DNA samples were tested for CNM and A. phagocytophilum by real-time PCR. Samples positive for A. phagocytophilum were further characterized at the 16S rRNA gene locus. CNM was detected in 28.6% of small mammals and in 2.2% of questing and 3.8% of attached ticks. Altogether 33 positive ticks were attached to 17 different hosts, while positive ticks per host ranged between one and seven. The prevalences for this pathogen differed significantly within small mammal populations comparing sites (χ(2): 13.3987; p: 0.0004) and between sexes. Male rodents had an approximately two times higher chance of infection than females (OR: 1.9652; 95% CI: 1.32-2.92). The prevalence for CNM was 31.8% (95% CI: 22-44) in rodent foetuses and neonates (23 of 67) from positive dams, and 60% (95% CI: 35.7-80.25) of positive gravid or recently parturient rodents (9 out of 15) had at least one positive foetus or neonate. Anaplasma phagocytophilum was detected at a low percentage in rodents (0-5.6%) and host-attached ticks (0.5-2.9%) with no significant differences between rodent species. However, attached nymphs were significantly more often infected than attached larvae (χ(2): 25.091; p: <0.0001). This study

  19. Dexamethasone-Induced Cytokine Changes Associated with Diminished Disease Severity in Horses Infected with Anaplasma phagocytophilum

    PubMed Central

    Davies, R. S.; Madigan, J. E.; Hodzic, E.; Borjesson, D. L; Dumler, J. S.

    2011-01-01

    Anaplasma phagocytophilum is the zoonotic cause of granulocytic anaplasmosis. We hypothesized that immune response, specifically gamma interferon (IFN-γ), plays a role in disease severity. To test this, horses were infected and IFNG expression was pharmacologically downregulated using corticosteroids. Eight horses were infected with A. phagocytophilum; 4 received dexamethasone on days 4 to 8 of infection. Clinical signs, hematologic parameters, and transcription of cytokine/chemokine genes were compared among treated and untreated horses. Infection was quantitated by msp2 real-time PCR and microscopy. As anticipated, there was significantly greater leukopenia, thrombocytopenia, and anemia in infected versus uninfected horses. The A. phagocytophilum load was higher for dexamethasone-treated horses. Dexamethasone reduced IFNG transcription by day 12 and IL-8 and IL-18 by days 7 to 9 and increased IL-4 on day 7. The ratio of IL-10 to IFNG was increased by dexamethasone on day 9. There were no hematologic differences between the infected horses. Dexamethasone suppression of proinflammatory response resulted in delayed infection-induced limb edema and decreased icterus, anorexia, and reluctance to move between days 6 and 9 and lower fever on day 7. These results underscore the utility of the equine model of granulocytic anaplasmosis and suggest that Th1 proinflammatory response plays a role in worsening disease severity and that disease severity can be decreased by modulating proinflammatory response. A role for Th1 response and macrophage activation in hematologic derangements elicited by A. phagocytophilum is not supported by these data and remains unproven. PMID:21880854

  20. Prevalence of antibodies against Borrelia burgdorferi, Anaplasma phagocytophilum, and Leptospira interrogans serovars in Bernese Mountain Dogs.

    PubMed

    Preyß-Jägeler, C; Müller, E; Straubinger, R K; Hartmann, K

    2016-01-01

    Bernese Mountain Dogs (BMD) have a higher prevalence of Borrelia burgdorferi sensu lato (Bbsl) antibodies than other breeds, but it is not known whether this is the case for other pathogens. Therefore, the aim of the study was to determine the frequency and level of specific antibodies against members of the Bbsl group, Anaplasma phagocytophilum (Ap), and Leptospira (L.) interrogans serovars in BMD and compare the results with those found in dogs of other breeds. A total of 171 healthy BMD and 57 healthy control dogs of other breeds were included in the study. Controls were large dogs (> 30 kg) with long, dark hair coats. A two-tiered testing method consisting of computerized kinetic enzyme-linked immunosorbent assay (KELA) and Western blotting was used for detection of antibodies against Bbsl, an immunofluorescence assay (IFA) was used for detection of antibodies against Ap, and microscopic agglutination test (MAT) for antibodies to 18 different serovars of L. interrogans. The prevalence of anti-Bbsl antibodies was significantly higher in BMD (43.3%) than in controls (17.5%) (p < 0.001). Antibodies to Bbsl attributable to vaccination were excluded from the calculation of prevalence. Antibodies to Ap were found in 50.3% of BMD, whereas only 24.6% of the controls dogs were tested positive for Ap (p < 0.001). Antibody titers of the 18 different serovars of L. interrogans antibodies did not differ significantly between BMD and control dogs except for L. copenhageni antibody titers which were higher in BMD. Significantly higher antibody titers to L. canicola (p = 0.003), L. copenhageni (p = 0.005), L. grippothyphosa (p = 0.029) and L. vanderhoedoni (p = 0.035) were seen in BMD compared to control dogs. BMD had a higher prevalence of anti-Bbsl, anti-L. copenhageni and anti-Ap antibodies than control dogs. Significantly higher antibody titers against L. canicola (p = 0.003), L. copenhageni (p = 0.005), L. grippothyphosa (p = 0.029) and L

  1. A loop-mediated isothermal amplification (LAMP) assay targeting 16S rRNA gene for rapid detection of Anaplasma phagocytophilum infection in sheep and goats.

    PubMed

    Ning, Changshen; Wang, Jinhong; Zhang, Yan; Wang, Xiaoxing; Cui, Yanyan; Yan, Yaqun; Wang, Rongjun; Jian, Fuchun; Zhang, Longxian

    2017-01-24

    Anaplasma phagocytophilum is a zoonotic pathogen and the causative agent of human granulocytic anaplasmosis (HGA) in humans and tick-borne fever in various kinds of animals. In the present study, a loop-mediated isothermal amplification (LAMP) assay for rapid detection of A. phagocytophilum was developed using primers specific to 16S rRNA gene of this organism. The LAMP assay was performed at 65 C for 60 min and terminated at 80 C for 10 min. The optimal reaction conditions, under which no cross-reaction was observed with other closely related tick borne parasites (Anaplasma bovis, Anaplasma ovis, Theileria luwenshuni, Babesia motasi and Schistosoma japonicum) was established. The assay exhibited much higher sensitivity when compared with conventional PCR (1 copy vs 1000 copies). To evaluate the applicability of the LAMP assay, 94 sheep field blood samples were analyzed for A. phagocytophilum infection using LAMP, nested PCR and conventional PCR assay at the same time. A positive LAMP result was obtained from 53 of the 94 samples (56.4%), while only 12 (12.8%) and 3 (3.2%) were tested positive by nested PCR and conventional PCR, respectively. In conclusion, this LAMP assay is a specific, sensitive, and rapid method for the detection of A. phagocytophilum in sheep.

  2. Molecular Survey of Anaplasma Species in Small Ruminants Reveals the Presence of Novel Strains Closely Related to A. phagocytophilum in Tunisia

    PubMed Central

    Said, Mourad Ben; Belkahia, Hanène; Zobba, Rosanna; Bousrih, Maha; Yahiaoui, Mouna; Daaloul-Jedidi, Monia; Mamlouk, Aymen; Gharbi, Mohamed; Messadi, Lilia

    2015-01-01

    Abstract A survey of Anaplasma species in small ruminants is still lacking in North African countries. In this study, the presence of A. phagocytophilum, A. phagocytophilum–related species, and A. ovis was investigated in a total of 563 healthy small ruminants (303 goats and 260 sheep), from 25 randomly selected flocks sampled in Tunisia. Anaplasma spp. and A. ovis overall infection rates were 95.0% and 93.8% in sheep and 69.6% and 65.3% in goats, respectively. A. phagocytophilum was not detected in any of tested animals. A total of 20 sheep (7.7%) and 144 goats (47.5%) were infected by Anaplasma strains genetically related to A. phagocytophilum. Both in sheep and goats A. ovis prevalence was higher in adults (≥2 years) than in young (<2 years) subjects (p = 0.001 and 0.002 for goats and sheep, respectively). In sheep, A. ovis prevalence was higher in ewes with respect to rams (p = 0.010). The A. ovis infection rate was significantly lower in goats of the local breed (p = 0.049) and it was higher in goats infested by ticks than in not infested animals (p = 0.005). Genetic analysis of the msp4 gene of A. ovis indicated the presence of strains shared by Tunisian sheep and goats. Sequence analysis and phylogenetic studies on the basis of the 16S rRNA gene provided evidence for the circulation of at least two different potentially novel species genetically related to A. phagocytophilum in Tunisian small ruminants. These findings cause concern about specificity of serological tests used for detection of A. phagocytophilum in ruminants and provide additional information for elucidating pathogenesis and molecular epidemiology of A. phagocytophilum and related species. PMID:26394065

  3. Detection of Borrelia lusitaniae, Rickettsia sp. IRS3, Rickettsia monacensis, and Anaplasma phagocytophilum in Ixodes ricinus collected in Madeira Island, Portugal.

    PubMed

    de Carvalho, Isabel Lopes; Milhano, Natacha; Santos, Ana Sofia; Almeida, Victor; Barros, Silvia C; De Sousa, Rita; Núncio, Maria Sofia

    2008-08-01

    A total of 300 Ixodes ricinus ticks were tested by polymerase chain reaction (PCR) for the presence of Borrelia spp., Rickettsia spp., and Anaplasma phagocytophilum. Sequence analysis demonstrated 8 (2.7%) ticks infected with B. lusitaniae, 60 (20%) with Rickettsia spp., and 1 (0.3%) with A. phagocytophilum. Seven (2.3%) ticks were coinfected with B. lusitaniae and Rickettsia spp., 2 (0.6%) with R. monacensis, and 5 (1.7%) with Rickettsia sp. IRS3. The results of this study suggest simultaneous transmission of multiple tick-borne agents on Madeira Island, Portugal.

  4. Rapid Sequential Changeover of Expressed p44 Genes during the Acute Phase of Anaplasma phagocytophilum Infection in Horses

    PubMed Central

    Wang, Xueqi; Rikihisa, Yasuko; Lai, Tzung-Hui; Kumagai, Yumi; Zhi, Ning; Reed, Stephen M.

    2004-01-01

    Anaplasma phagocytophilum immunodominant polymorphic major surface protein P44s have been hypothesized to go through antigenic variation, but the within-host dynamics of p44 expression has not been demonstrated. In the present study we investigated the composition and changes of p44 transcripts in the blood during the acute phase of well-defined laboratory A. phagocytophilum infections in naïve equine hosts. Three traveling waves of sequential population changeovers of the p44 transcript species were observed within a single peak of rickettsemia of less than 1 month. During the logarithmic increase, the rapid switch-off of the initial dominant transcript p44-18 occurred regardless of whether the bacterium was transmitted by ticks or by intravenous inoculation. Each of the subsequently dominant p44 transcript species was phylogenetically dissimilar from p44-18. Development of antibody to the hypervariable region of P44-18 during the rickettsemia suggests the suppression of dominance of immuno-cross-reactive p44 populations. When A. phagocytophilum was preincubated with plasma from the infected horse and then coincubated with HL-60 cells, the dominance of the p44-18 transcript was rapidly suppressed in vitro and most of the newly emerged p44 transcript species were previously undetected in this horse. This work provides experimental evidence of within-host p44 antigenic variation. Results suggest that the rapid and synchronized switch of expression is an intrinsic property of p44s reinitiated after transmission to naïve mammalian hosts and shaped upon exposure to immune plasma. PMID:15557606

  5. IL-12/23p40-dependent clearance of Anaplasma phagocytophilum in the murine model of human anaplasmosis.

    PubMed

    Pedra, Joao H F; Tao, Jian; Sutterwala, Fayyaz S; Sukumaran, Bindu; Berliner, Nancy; Bockenstedt, Linda K; Flavell, Richard A; Yin, Zhinan; Fikrig, Erol

    2007-08-01

    Human anaplasmosis is an emerging infectious disease transmitted by ticks that can be potentially fatal in the immunocompromised and the elderly. The mechanisms of defense against the causative agent, Anaplasma phagocytophilum, are not completely understood; however, interferon (IFN)-gamma plays an important role in pathogen clearance. Here, we show that IFN-gamma is regulated through an early IL-12/23p40-dependent mechanism. Interleukin (IL)-12/23p40 is regulated in macrophages and dendritic cells after activation by microbial agonists and cytokines and constitutes a subunit of IL-12 and IL-23. IL-12/23p40-deficient mice displayed an increased A. phagocytophilum burden, accelerated thrombocytopenia and increased neutrophil numbers in the spleen at day 6 postinfection. Infection of MyD88- and mitogen-activated kinase kinase 3 (MKK3)-deficient mice suggested that the early susceptibility due to IL-12/23p40 deficiency was not dependent on signaling through MyD88 or MKK3. The lack of IL-12/23p40 reduced IFN-gamma production in both CD4(+) and CD8(+) T cells although the effect was more pronounced in CD4(+) T cells. Our data suggest that the immune response against A. phagocytophilum is a multifactorial and cooperative process. The IL-12/23p40 subunit drives the CD4(+) Th1 immune response in the early phase of infection and IL-12/23p40-independent mechanisms ultimately contribute to pathogen elimination from the host.

  6. Northern white-breasted hedgehogs Erinaceus roumanicus as hosts for ticks infected with Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum in Romania.

    PubMed

    Dumitrache, Mirabela Oana; Paştiu, Anamaria Ioana; Kalmár, Zsuzsa; Mircean, Viorica; Sándor, Attila David; Gherman, Călin Mircea; Peştean, Cosmin; Mihalca, Andrei Daniel; Cozma, Vasile

    2013-04-01

    Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum are two tick-borne pathogens of medical and/or veterinary importance which are distributed worldwide. Erinaceus roumanicus, the Northern white-breasted hedgehog, is a common synanthropic species that is known to carry not only the hedgehog tick, Ixodes hexagonus, but also I. ricinus, the most common European tick species. I. ricinus is the main vector of both mentioned pathogens. Within this framework and because only limited information is available on the role of E. roumanicus in the ecology of B. burgdorferi s.l. and Anaplasma phagocytophilum in Europe, we carried out an epidemiological surveillance on this species in Romania. From the 57 examined hedgehogs collected in 12 different counties, 24 presented tick infestation. Most ticks (n=959) were morphologically identified as larvae, nymphs, or adults of I. ricinus. The prevalence of B. burgdorferi s.l. was 0.4%, and that of A. phagocytophilum 12%. In all positive cases for B. burgdorferi s.l., restriction fragment length polymorphism revealed the genospecies B. afzelii. In Romania, only limited information is available on the epidemiology of B. burgdorferi s.l. and A. phagocytophilum. As hedgehogs commonly share the same environment with humans and other potential reservoir hosts for tick-borne pathogens, our study provides new epidemiological data of public health importance.

  7. Molecular Analysis of Anaplasma phagocytophilum Isolated from Patients with Febrile Diseases of Unknown Etiology in China

    PubMed Central

    Li, Jun; Long, Bo; Yu, Hong; Zhang, Zhilun; He, Jing; Qu, Zhangyi; Yu, Jiguang; Liu, Yuanni; Dong, Tuo; Yao, Na; Wang, Yong; Cheng, Xueqin; Xu, Jianguo

    2013-01-01

    Although anaplasmosis cases have been nationally identified in China, no human isolates of A. phagocytophilum have been obtained, which limits the analysis of any molecular and genetic contributions to patients' severe clinical manifestations and the study of the bacteria's pathogeneses in China. Given this situation, a joint project was conducted in 2009–2010. A total of 421 febrile cases of unknown etiology were collected and the patients' blood samples were collected for laboratory diagnoses including serologic diagnosis based on the four-fold rise in the anti- A. phagocytophilum IgG titer by indirect micro-immunofluorescence assay (IFA), positive PCR assay and confirmation of A. phagocytophilum DNA and positive culture of A. phagocytophilum and confirmed by amplification and sequencing of the 16S rRNA and ank A genes of the A. phagocytophilum isolates. A total of 570 ticks were collected from the patients' domestic animals (456) and from wild fields (114) for culturing and amplifying and sequencing the 16S rRNA gene of A. phagocytophilum. Phylogenetic analyses were performed on the 16S rRNA and ank A gene sequences of the isolates and the ticks tested in the study. A total of 46 (10.9%) confirmed and 16 (3.8%) probable cases were diagnosed and severe clinical features and higher mortality rates were observed in these Chinese patients. Five isolates were obtained and the 16S rRNA genes of the 5 isolates were conserved but variety for ank A genes. Two human isolates and 1 tick isolate from Shandong Peninsula, where all patients exhibited severe clinical manifestations, were grouped as one clan based on the phylogenetic analyses, while 2 other human isolates were clustered in a second clan. 43.5% of H. longicornis were infected with A. phagocytophilum.The present study is the first to obtain clinical isolates of A. phagocytophilum in China. The diversity of the ank A genes of Chinese isolates will help us to further discern the relationship between the

  8. Efficacy of sarolaner in the prevention of Borrelia burgdorferi and Anaplasma phagocytophilum transmission from infected Ixodes scapularis to dogs.

    PubMed

    Honsberger, Nicole A; Six, Robert H; Heinz, Thomas J; Weber, Angela; Mahabir, Sean P; Berg, Thomas C

    2016-05-30

    The efficacy of sarolaner (Simparica™, Zoetis) to prevent transmission primarily of Borrelia burgdorferi and secondarily of Anaplasma phagocytophilum from infected wild-caught Ixodes scapularis to dogs was evaluated in a placebo-controlled laboratory study. Twenty-four purpose-bred laboratory Beagles seronegative for B. burgdorferi and A. phagocytophilum antibodies were allocated randomly to one of three treatment groups: placebo administered orally on Days 0 and 7, or sarolaner at 2mg/kg administered orally on Day 0 (28 days prior to tick infestation) or on Day 7 (21 days prior to tick infestation). On Day 28, each dog was infested with approximately 25 female and 25 male wild caught adult I. scapularis that were determined to have prevalence of 57% for B. burgdorferi and 6.7% for A. phagocytophilum by PCR. In situ tick counts were conducted on Days 29 and 30. On Day 33, all ticks were counted and removed. Acaricidal efficacy was calculated based on the reduction of geometric mean live tick counts in the sarolaner-treated groups compared to the placebo-treated group for each tick count. Blood samples collected from each dog on Days 27, 49, 63, 77, 91 and 104 were tested for the presence of B. burgdorferi and A. phagocytophilum antibodies using the SNAP(®) 4Dx(®) Plus Test, and quantitatively assayed for B. burgdorferi antibodies using an ELISA test. Skin biopsies collected on Day 104 were tested for the presence of B. burgdorferi by bacterial culture and PCR. Geometric mean live tick counts for placebo-treated dogs were 14.8, 12.8, and 19.1 on Days 29, 30, and 33, respectively. The percent reductions in mean live tick counts at 1, 2, and 5 days after infestation were 86.3%, 100%, and 100% for the group treated with sarolaner 21 days prior to infestation, and 90.9%, 97.1%, and 100% for the group treated with sarolaner 28 days prior to infestation. Geometric mean live tick counts for both sarolaner-treated groups were significantly lower than those for the

  9. Comparative Experimental Infection Study in Dogs with Ehrlichia canis, E. chaffeensis, Anaplasma platys and A. phagocytophilum.

    PubMed

    Nair, Arathy D S; Cheng, Chuanmin; Ganta, Chanran K; Sanderson, Michael W; Alleman, Arthur R; Munderloh, Ulrike G; Ganta, Roman R

    2016-01-01

    Dogs acquire infections with the Anaplasmataceae family pathogens, E. canis, E. chaffeensis, E. ewingii, A. platys and A. phagocytophilum mostly during summer months when ticks are actively feeding on animals. These pathogens are also identified as causing diseases in people. Despite the long history of tick-borne diseases in dogs, much remains to be defined pertaining to the clinical and pathological outcomes of infections with these pathogens. In the current study, we performed experimental infections in dogs with E. canis, E. chaffeensis, A. platys and A. phagocytophilum. Animals were monitored for 42 days to evaluate infection-specific clinical, hematological and pathological differences. All four pathogens caused systemic persistent infections detectible throughout the 6 weeks of infection assessment. Fever was frequently detected in animals infected with E. canis, E. chaffeensis, and A. platys, but not in dogs infected with A. phagocytophilum. Hematological differences were evident in all four infected groups, although significant overlap existed between the groups. A marked reduction in packed cell volume that correlated with reduced erythrocytes and hemoglobin was observed only in E. canis infected animals. A decline in platelet numbers was common with E. canis, A. platys and A. phagocytophilum infections. Histopathological lesions in lung, liver and spleen were observed in all four groups of infected dogs; infection with E. canis had the highest pathological scores, followed by E. chaffeensis, then A. platys and A. phagocytophilum. All four pathogens induced IgG responses starting on day 7 post infection, which was predominantly comprised of IgG2 subclass antibodies. This is the first detailed investigation comparing the infection progression and host responses in dogs after inoculation with four pathogens belonging to the Anaplasmataceae family. The study revealed a significant overlap in clinical, hematological and pathological changes resulting from the

  10. Comparative Experimental Infection Study in Dogs with Ehrlichia canis, E. chaffeensis, Anaplasma platys and A. phagocytophilum

    PubMed Central

    Nair, Arathy D. S.; Cheng, Chuanmin; Ganta, Chanran K.; Sanderson, Michael W; Alleman, Arthur R.; Munderloh, Ulrike G.; Ganta, Roman R.

    2016-01-01

    Dogs acquire infections with the Anaplasmataceae family pathogens, E. canis, E. chaffeensis, E. ewingii, A. platys and A. phagocytophilum mostly during summer months when ticks are actively feeding on animals. These pathogens are also identified as causing diseases in people. Despite the long history of tick-borne diseases in dogs, much remains to be defined pertaining to the clinical and pathological outcomes of infections with these pathogens. In the current study, we performed experimental infections in dogs with E. canis, E. chaffeensis, A. platys and A. phagocytophilum. Animals were monitored for 42 days to evaluate infection-specific clinical, hematological and pathological differences. All four pathogens caused systemic persistent infections detectible throughout the 6 weeks of infection assessment. Fever was frequently detected in animals infected with E. canis, E. chaffeensis, and A. platys, but not in dogs infected with A. phagocytophilum. Hematological differences were evident in all four infected groups, although significant overlap existed between the groups. A marked reduction in packed cell volume that correlated with reduced erythrocytes and hemoglobin was observed only in E. canis infected animals. A decline in platelet numbers was common with E. canis, A. platys and A. phagocytophilum infections. Histopathological lesions in lung, liver and spleen were observed in all four groups of infected dogs; infection with E. canis had the highest pathological scores, followed by E. chaffeensis, then A. platys and A. phagocytophilum. All four pathogens induced IgG responses starting on day 7 post infection, which was predominantly comprised of IgG2 subclass antibodies. This is the first detailed investigation comparing the infection progression and host responses in dogs after inoculation with four pathogens belonging to the Anaplasmataceae family. The study revealed a significant overlap in clinical, hematological and pathological changes resulting from the

  11. Prevalence of antibodies against Rickettsia conorii, Babesia canis, Ehrlichia canis, and Anaplasma phagocytophilum antigens in dogs from the Stretto di Messina area (Italy).

    PubMed

    Pennisi, Maria-Grazia; Caprì, Alessandra; Solano-Gallego, Laia; Lombardo, Gabriella; Torina, Alessandra; Masucci, Marisa

    2012-12-01

    The aims of this study were to determine the seroprevalence for Rickettsia conorii, Ehrlichia canis, Anaplasma phagocytophilum, and Babesia canis in outdoor-kennelled dogs (n=249) from the Stretto di Messina (Italy) and to compare seroprevalence in 2 public shelters and 4 privately-owned kennels where different tick-preventive measures were implemented in order to focus on the specific sanitary risk posed by public shelters in southern Italy for tick-borne pathogens. R. conorii (72%) and B. canis (70%) were the most prevalent infections when compared to E. canis (46%) and A. phagocytophilum (38%). Seroprevalence for R. conorii, E. canis, and A. phagocytophilum was significantly higher in public shelters than in private kennels. However, B. canis seropositivity was similar in both types of kennels. In addition, in private kennels where a regular ectocide treatment was carried out by means of spot-on devices, dogs did not present E. canis and A. phagocytophilum antibodies. One hundred fifty-one dogs out of 249 (61%) were seropositive to more than one pathogen with R. conorii and B. canis the most common ones. Coinfections were more frequently found in public-shelter dogs. This study demonstrated high seroprevalences against R. conorii, B. canis, E. canis, and A. phagocytophilum in kennelled dogs from both coastal sites of the Stretto di Messina and the importance of regular tick-bite prevention by means of individual spot-on devices.

  12. Coinfection of western gray squirrel (Sciurus griseus) and other sciurid rodents with Borrelia burgdorferi sensu stricto and Anaplasma phagocytophilum in California.

    PubMed

    Nieto, Nathan C; Leonhard, Sarah; Foley, Janet E; Lane, Robert S

    2010-01-01

    Overlapping geographic distributions of tick-borne disease agents utilizing the same tick vectors are common, and coinfection of humans, domestic animals, wildlife, and ticks with both Borrelia burgdorferi and Anaplasma phagocytophilum has been frequently reported. This study was undertaken in order to evaluate the prevalence of both B. burgdorferi sensu stricto (hereinafter referred to as B. burgdorferi) and A. phagocytophilum in several species of sciurid rodents from northern California, USA. Rodents were either collected dead as road-kills or live-trapped in four state parks from 13 counties. Thirty-seven western gray squirrels (Sciurus griseus), nine nonnative eastern gray squirrels (S. carolinensis) and an eastern fox squirrel (S. niger), four Douglas squirrels (Tamiasciurus douglasii), and two northern flying squirrels (Glaucomys sabrinus) were tested by polymerase chain reaction (PCR) and serology for evidence of coinfection. Of the 14 individual S. griseus that were PCR-positive for B. burgdorferi, two (14%) also were PCR-positive for A. phagocytophilum and 11 (79%) had serologic evidence of A. phagocytophilum exposure. Two of the four Douglas squirrels were PCR positive for B. burgdorferi and seropositive to A. phagocytophilum. Evidence of coinfection with these zoonotic pathogens in western gray squirrels suggests that both bacteria may be maintained in a similar transmission cycle involving this sciurid and the western black-legged tick Ixodes pacificus, the primary bridging vector to humans in the far-western US.

  13. Detection and characterization of p44/msp2 transcript variants of Anaplasma phagocytophilum from naturally infected ticks and wild deer in Japan.

    PubMed

    Gaowa; Wuritu; Wu, Dongxing; Yoshikawa, Yuko; Ohashi, Noiro; Kawamori, Fumihiko; Sugiyama, Kanji; Ohtake, Masayoshi; Ohashi, Masataka; Yamamoto, Seigo; Kitano, Tomokazu; Takada, Nobuhiro; Kawabata, Hiroki

    2012-01-01

    Anaplasma phagocytophilum is an obligate intracellular bacterium and causes a febrile illness in humans and livestock. In nature, this bacterium is sustained in a tick-mammal cycle. Several p44/msp2-related genes are expressed from a single expression locus by gene conversion. In this study, we obtained 119 cDNA sequences of p44/msp2 transcripts from A. phagocytophilum in 6 Haemaphysalis ticks and 3 wild sika deer (Cervus nippon) in Japan. These 119 sequences were classified into 36 different variant sequences based on their similarities. The 36 cDNA sequences were phylogenetically grouped into 2 major clusters--tick- and deer-associated. The tick-associated sequences were further classified into 4 distinct subclusters, suggesting that A. phagocytophilum in ticks seems to selectively express specific p44/msp2 transcripts, such as the transcripts in the 4 subclusters that were closely related to previously identified p44/msp2 genes. The deer-associated sequences were also grouped into 4 subclusters, but these transcripts were probably more diverse than the transcripts derived from ticks. This might be due to the relatively nonselective expression of p44/msp2 in deer or the strain differences in A. phagocytophilum from ticks and deer in separate geographic regions or both. Thus, this study may contribute to the understanding of A. phagocytophilum p44/msp2 expression in nature in Japan.

  14. High-resolution genetic fingerprinting of European strains of Anaplasma phagocytophilum by use of multilocus variable-number tandem-repeat analysis.

    PubMed

    Bown, Kevin J; Lambin, Xavier; Ogden, Nicholas H; Petrovec, Miroslav; Shaw, Susan E; Woldehiwet, Zerai; Birtles, Richard J

    2007-06-01

    Anaplasma phagocytophilum is a widely distributed tick-borne pathogen of humans, livestock, and companion animals. We used in silico methods to identify 10 variable-number tandem-repeat (VNTR) loci within the genome sequence of the A. phagocytophilum HZ strain and used these data to develop a multilocus VNTR-based typing scheme for the species. Having confirmed the stability of four of the loci in replicates of the A. phagocytophilum strain that had been subjected to different numbers of passages through cell cocultures in vitro, we then used this typing scheme to discriminate between 20 A. phagocytophilum strains of diverse geographical and host provenances. Extensive diversity was found at each of the four loci studied, with total allele numbers ranging from 13 to 18 and Hunter-Gaston discriminatory index values ranging from 0.93 to 0.99. Only 2 of the 20 strains examined shared alleles at all four loci. The discriminatory power of VNTR analysis was found to be greater than that of either partial msp4 or 16S rRNA gene sequence comparison. The extremely high sensitivity of this novel approach to the genetic fingerprinting of A. phagocytophilum strains should serve well in molecular epidemiological studies of infection transmission, particularly when fine-scale strain delineation is required.

  15. Intra-leukocyte expression of two-component systems in Ehrlichia chaffeensis and Anaplasma phagocytophilum and effects of the histidine kinase inhibitor closantel.

    PubMed

    Cheng, Zhihui; Kumagai, Yumi; Lin, Mingqun; Zhang, Chunbin; Rikihisa, Yasuko

    2006-08-01

    The two-component system (TCS) composed of a pair of a sensor histidine kinase and a response regulator, allows bacteria to sense signals and respond to changes in their environment through specific gene activation or repression. The present study examined TCS in the obligatory intracellular bacteria Ehrlichia chaffeensis and Anaplasma phagocytophilum, that cause human monocytic ehrlichiosis (HME) and human granulocytic anaplasmosis (HGA) respectively. The genomes of E. chaffeensis and A. phagocytophilum were each predicted to encode three pairs of TCSs. All six genes encoding three histidine kinases and three response regulators were expressed in both E. chaffeensis and A. phagocytophilum cultured in human leukocytes. Pretreatment of host cell-free E. chaffeensis or A. phagocytophilum with closantel, an inhibitor of histidine kinases, completely blocked the infection of host cells. Treatment of infected cells 1 day post infection with closantel cleared infection in dose-dependent manner. All six genes in E. chaffeensis were cloned, recombinant proteins were expressed, and polyclonal antibodies were produced. Double immunofluorescence labelling and Western blot analysis revealed that all six proteins were expressed in cell culture. Autokinase activities of the three recombinant histidine kinases from E. chaffeensis were inhibited by closantel in vitro. A number of E. chaffeensis genes, including the six TCS genes, were downregulated within 5-60 min post closantel treatment. These results suggest that these TCSs play an essential role in infection and survival of E. chaffeensis and A. phagocytophilum in human leukocytes.

  16. Babesia spp. and Anaplasma phagocytophilum in questing ticks, ticks parasitizing rodents and the parasitized rodents – Analyzing the host-pathogen-vector interface in a metropolitan area

    PubMed Central

    2012-01-01

    Background The aims of this study were to evaluate the host-tick-pathogen interface of Babesia spp. and Anaplasma phagocytophilum in restored areas in both questing and host-attached Ixodes ricinus and Dermacentor reticulatus and their small mammalian hosts. Methods Questing ticks were collected from 5 sites within the city of Leipzig, Germany, in 2009. Small mammals were trapped at 3 of the 5 sites during 2010 and 2011. DNA extracts of questing and host-attached I. ricinus and D. reticulatus and of several tissue types of small mammals (the majority bank voles and yellow-necked mice), were investigated by PCR followed by sequencing for the occurrence of DNA of Babesia spp. and by real-time PCR for A. phagocytophilum. A selected number of samples positive for A. phagocytophilum were further investigated for variants of the partial 16S rRNA gene. Co-infection with Rickettsia spp. in the questing ticks was additionally investigated. Results 4.1% of questing I. ricinus ticks, but no D. reticulatus, were positive for Babesia sp. and 8.7% of I. ricinus for A. phagocytophilum. Sequencing revealed B. microti, B. capreoli and Babesia spp. EU1 in Leipzig and sequence analysis of the partial 16S RNA gene of A. phagocytophilum revealed variants either rarely reported in human cases or associated with cervid hosts. The statistical analysis revealed significantly less ticks infected with A. phagocytophilum in a city park in Leipzig as compared to the other sampling sites. A. phagocytophilum-DNA was detected in 2 bank voles, DNA of B. microti in 1 striped field-mouse and of Babesia sp. EU1 in the skin tissue of a mole. Co-infections were detected. Conclusion Our results show the involvement of small mammals in the natural endemic cycles of tick-borne pathogens. A more thorough understanding of the interactions of ticks, pathogens and hosts is the essential basis for effective preventive control measures. PMID:22950642

  17. Genetic variability of Anaplasma phagocytophilum in ticks and voles from Ixodes persulcatus/Ixodes trianguliceps sympatric areas from Western Siberia, Russia.

    PubMed

    Rar, Vera A; Epikhina, Tamara I; Yakimenko, Valeriy V; Malkova, Marina G; Tancev, Aleksey K; Bondarenko, Evgeny I; Ivanov, Mikhail K; Tikunova, Nina V

    2014-10-01

    Anaplasma phagocytophilum is a causative agent of granulocytic anaplasmosis in different mammals. The presence of A. phagocytophilum was assayed in Ixodes persulcatus, Ixodes trianguliceps ticks and Myodes spp. voles from two I. persulcatus/I. trianguliceps sympatric areas in the Omsk region (Western Siberia, Russia). In total, A. phagocytophilum was found in 42/108 (38.9%) of vole blood samples, 13/34 (38.2%) of I. trianguliceps ticks removed from voles, 1/12 (8.3%) of I. persulcatus removed from voles, and 18/279 (7.2%) of questing I. persulcatus. GroESL operon sequence analysis of positive samples revealed three distinct A. phagocytophilum genetic groups previously identified in ticks and mammals in Russia. Genetic group 1 was found in 6/36 (16.7%) of sequenced positive blood samples; this group was previously revealed in I. persulcatus and Myodes spp. voles in different regions of Russia. Genetic group 2 was found in 30/36 (83.3%) of sequenced positive blood samples and all positive I. trianguliceps; this group was previously revealed only in Myodes spp. voles and common shrews (Sorex araneus) in I. persulcatus/I. trianguliceps sympatric areas in the Northern Ural. Genetic group 3 was found in all positive questing I. persulcatus and one blood sample; this group was previously revealed in I. persulcatus and Siberian chipmunks (Tamias sibiricus). We suppose that I. trianguliceps is the most probable vector for A. phagocytophilum of group 2. Analysis of the msp4 gene, intergenic region DOV1, and some other genetic loci has shown that isolates from different genetic groups significantly differ in all studied loci and that A. phagocytophilum of group 2 is closely related to A. phagocytophilum isolates revealed in voles and I. trianguliceps in Europe. A. phagocytophilum of groups 1 and 2 are the most similar to each other, while A. phagocytophilum of group 3 clusters with European A. phagocytophilum isolates from I. ricinus and various mammalian species. Copyright

  18. Transovarial Transmission of Francisella-Like Endosymbionts and Anaplasma phagocytophilum Variants in Dermacentor albipictus (Acari: Ixodidae)

    USDA-ARS?s Scientific Manuscript database

    Dermacentor albipictus (Packard) is a North American Ixodid tick that parasitizes deer species but also infests livestock. It is a suspected vector of the agent of anaplasmosis in cattle herds, Anaplasma marginale, but its microbial flora and emerging pathogen vector potential remain under-evaluated...

  19. Surveillance for Ixodes pacificus and the tick-borne pathogens Anaplasma phagocytophilum and Borrelia burgdorferi in birds from California's Inner Coast Range.

    PubMed

    Dingler, Regina J; Wright, Stan A; Donohue, Ann M; Macedo, Paula A; Foley, Janet E

    2014-06-01

    We investigated the involvement of birds in the ecology of the western black-legged tick, Ixodes pacificus, and its associated zoonotic bacteria, Borrelia burgdorferi and Anaplasma phagocytophilum, at two interior coast-range study sites in northern California. Anaplasma phagocytophilum, the agent of granulocytic anaplasmosis (GA), and B. burgdorferi s.s., the agent of Lyme disease (LD), are tick-borne pathogens that are well established in California. We screened blood and ticks from 349 individual birds in 48 species collected in 2011 and 2012 using pathogen-specific PCR. A total of 617 immature I. pacificus was collected with almost three times as many larvae than nymphs. There were 7.5 times more I. pacificus at the Napa County site compared to the Yolo County site. Two of 74 (3%) nymphal pools from an Oregon junco (Junco hyemalis) and a hermit thrush (Catharus guttatus) and 4 individual larvae (all from Oregon juncos) were PCR-positive for B. burgdorferi. Blood samples from a golden-crowned sparrow (Zonotrichia atricapilla) and a European starling (Sturnus vulgaris) were positive for A. phagocytophilum DNA at very low levels. Birds that forage on ground or bark and nest on the ground, as well as some migratory species, are at an increased risk for acquiring I. pacificus. Our findings show that birds contribute to the ecologies of LD and GA in California by serving as a blood-meal source, feeding and transporting immature I. pacificus, and sometimes as a source of Borrelia infection.

  20. Borrelia burgdorferi sensu lato and co-infections with Anaplasma phagocytophilum and Rickettsia spp. in Ixodes ricinus in Hamburg, Germany.

    PubMed

    May, K; Jordan, D; Fingerle, V; Strube, C

    2015-12-01

    To obtain initial data on Borrelia burgdorferi sensu lato (Spirochaetales: Spirochaetaceae) in Ixodes ricinus (Ixodida: Ixodidae) ticks in Hamburg, Germany, 1400 questing ticks were collected by flagging at 10 different public recreation areas in 2011 and analysed using probe-based quantitative real-time polymerase chain reaction. The overall rate of infection with B. burgdorferi s.l. was 34.1%; 30.0% of adults were infected (36.7% of females and 26.0% of males), as were 34.5% of nymphs. Significant differences in tick infection rates were observed between the spring and summer/autumn months, as well as among sampling locations. Borrelia genospecies identification by reverse line blotting was successful in 43.6% of positive tick samples. The most frequent genospecies was Borrelia garinii/Borrelia bavariensis, followed by Borrelia afzelii, Borrelia valaisiana, B. burgdorferi sensu stricto, Borrelia spielmanii, Borrelia bissettii and Borrelia lusitaniae. Based on previously published data, co-infection of Borrelia and Rickettsiales spp. was determined in 25.8% of ticks. Overall, 22.9% of ticks were co-infected with Rickettsia spp. (Rickettsiales: Rickettsiaceae), 1.7% with Anaplasma phagocytophilum (Rickettsiales: Anaplasmataceae), and 1.2% with both pathogens. Study results show a high prevalence of Borrelia-positive ticks in recreation areas in the northern German city of Hamburg and the potential health risk to humans in these areas should not be underestimated.

  1. Serological reactivity to Ehrlichia canis, Anaplasma phagocytophilum, Neorickettsia risticii, Borrelia burgdorferi and Rickettsia conorii in dogs from northwestern Spain.

    PubMed

    Amusategui, Inmaculada; Tesouro, Miguel A; Kakoma, Ibulaimu; Sainz, Angel

    2008-12-01

    The aim of the present work was to investigate the seroprevalence against Ehrlichia canis (Ec), Anaplasma phagocytophilum (Ap), Neorickettsia risticii (Nr), Rickettsia conorii (Rc), and Borrelia burgdorferi (Bb) in two different clusters of canine samples from Northwestern Spain. Cluster 1 included 479 dogs presented at veterinary clinics located in Ourense and Pontevedra. Cluster II included 170 dogs from the public kennel of Ourense. All 649 canine serum samples were analyzed by immunofluorescent antibody test. Prevalences against the above-mentioned agents in cluster I were: Rc (24.6%), Bb (6.26%), Ec (3.13%), Ap (5.01%), and Nr (1.04%), whereas for cluster II were: Rc (50%), Bb (8.8%), Ec (54.7%), Ap (45.3%), and Nr (4.7%). Rc was significantly associated with age and history of exposure to ticks, and Bb showed a statistical relationship with age and clinical status. Ec and Ap were related to the occupation of the dogs, with stray dogs being the most frequently seropositive. Furthermore, seroreactivity against Ec and Ap was significantly higher in Ourense than in Pontevedra. The univariate analysis demonstrated a significant concomitant seroreactivity between Ec and Ap and between Rc and Ec and Ap antigens. The seroreactivity to Nr must be interpreted very cautiously as this infectious agent has been seldom reported outside North America.

  2. Seroprevalence and geographic distribution of Dirofilaria immitis and tick-borne infections (Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, and Ehrlichia canis) in dogs from Romania.

    PubMed

    Mircean, Viorica; Dumitrache, Mirabela Oana; Györke, Adriana; Pantchev, Nikola; Jodies, Robert; Mihalca, Andrei Daniel; Cozma, Vasile

    2012-07-01

    Tick-borne diseases are of great concern worldwide. Despite this, in Romania there is only limited information regarding the prevalence of vector-borne pathogens in dogs. In all, 1146 serum samples were tested by SNAP(®) 4Dx(®) (IDEXX Laboratories, Inc., Westbrook, ME) for Anaplasma phagocytophilum, Borrelia burgdorferi, and Ehrlichia canis antibodies, and for Dirofilaria immitis antigen. The correlation between positive cases and their geographic distribution, as well as potential risk factors (age, sex, breed, type of dog, habitat, and prophylactic treatments) were evaluated. Overall, 129 dogs (11.3%) were serologically-positive to one or more of the tested pathogens. The seroprevalence for the four infectious agents were: A. phagocytophilum 5.5% (63/1146), D. immitis 3.3% (38/1146), E. canis 2.1% (24/1146), and B. burgdorferi 0.5% (6/1146). Co-infection with E. canis and A. phagocytophilum was registered in 2 dogs (0.2%). The geographical distribution of the seropositive cases suggests clustered foci in southern regions and in the western part of the country for D. immitis, and in the southeastern region (Constanţa County) for E. canis. A. phagocytophilum and B. burgdorferi showed a homogenous distribution, with a tendency for Lyme-positive samples to concentrate in central Romania. For D. immitis, A. phagocytophilum, and E. canis, administering prophylactic treatments was a risk factor associated with infection. Another associated risk factor was the type of dog (stray dogs were at risk being positive for D. immitis, shelter dogs for E. canis, and hunting dogs for B. burgdorferi). The prevalence of D. immitis was significantly higher in males and in dogs older than 2 years. This survey represents the first data detailing A. phagocytophilum and E. canis seroprevalence in Romanian dogs, and the most comprehensive epidemiological study on vector-borne infections in dogs from this country.

  3. Tissue-Specific Signatures in the Transcriptional Response to Anaplasma phagocytophilum Infection of Ixodes scapularis and Ixodes ricinus Tick Cell Lines

    PubMed Central

    Alberdi, Pilar; Mansfield, Karen L.; Manzano-Román, Raúl; Cook, Charlotte; Ayllón, Nieves; Villar, Margarita; Johnson, Nicholas; Fooks, Anthony R.; de la Fuente, José

    2016-01-01

    Anaplasma phagocytophilum are transmitted by Ixodes spp. ticks and have become one of the most common and relevant tick-borne pathogens due to their impact on human and animal health. Recent results have increased our understanding of the molecular interactions between Ixodes scapularis and A. phagocytophilum through the demonstration of tissue-specific molecular pathways that ensure pathogen infection, development and transmission by ticks. However, little is known about the Ixodes ricinus genes and proteins involved in the response to A. phagocytophilum infection. The tick species I. scapularis and I. ricinus are evolutionarily closely related and therefore similar responses are expected in A. phagocytophilum-infected cells. However, differences may exist between I. scapularis ISE6 and I. ricinus IRE/CTVM20 tick cells associated with tissue-specific signatures of these cell lines. To address this hypothesis, the transcriptional response to A. phagocytophilum infection was characterized by RNA sequencing and compared between I. scapularis ISE6 and I. ricinus IRE/CTVM20 tick cell lines. The transcriptional response to infection of I. scapularis ISE6 cells resembled that of tick hemocytes while the response in I. ricinus IRE/CTVM20 cells was more closely related to that reported previously in infected tick midguts. The inhibition of cell apoptosis by A. phagocytophilum appears to be a key adaptation mechanism to facilitate infection of both vertebrate and tick cells and was used to investigate further the tissue-specific response of tick cell lines to pathogen infection. The results supported a role for the intrinsic pathway in the inhibition of cell apoptosis by A. phagocytophilum infection of I. scapularis ISE6 cells. In contrast, the results in I. ricinus IRE/CTVM20 cells were similar to those obtained in tick midguts and suggested a role for the JAK/STAT pathway in the inhibition of apoptosis in tick cells infected with A. phagocytophilum. Nevertheless, tick

  4. Tissue-Specific Signatures in the Transcriptional Response to Anaplasma phagocytophilum Infection of Ixodes scapularis and Ixodes ricinus Tick Cell Lines.

    PubMed

    Alberdi, Pilar; Mansfield, Karen L; Manzano-Román, Raúl; Cook, Charlotte; Ayllón, Nieves; Villar, Margarita; Johnson, Nicholas; Fooks, Anthony R; de la Fuente, José

    2016-01-01

    Anaplasma phagocytophilum are transmitted by Ixodes spp. ticks and have become one of the most common and relevant tick-borne pathogens due to their impact on human and animal health. Recent results have increased our understanding of the molecular interactions between Ixodes scapularis and A. phagocytophilum through the demonstration of tissue-specific molecular pathways that ensure pathogen infection, development and transmission by ticks. However, little is known about the Ixodes ricinus genes and proteins involved in the response to A. phagocytophilum infection. The tick species I. scapularis and I. ricinus are evolutionarily closely related and therefore similar responses are expected in A. phagocytophilum-infected cells. However, differences may exist between I. scapularis ISE6 and I. ricinus IRE/CTVM20 tick cells associated with tissue-specific signatures of these cell lines. To address this hypothesis, the transcriptional response to A. phagocytophilum infection was characterized by RNA sequencing and compared between I. scapularis ISE6 and I. ricinus IRE/CTVM20 tick cell lines. The transcriptional response to infection of I. scapularis ISE6 cells resembled that of tick hemocytes while the response in I. ricinus IRE/CTVM20 cells was more closely related to that reported previously in infected tick midguts. The inhibition of cell apoptosis by A. phagocytophilum appears to be a key adaptation mechanism to facilitate infection of both vertebrate and tick cells and was used to investigate further the tissue-specific response of tick cell lines to pathogen infection. The results supported a role for the intrinsic pathway in the inhibition of cell apoptosis by A. phagocytophilum infection of I. scapularis ISE6 cells. In contrast, the results in I. ricinus IRE/CTVM20 cells were similar to those obtained in tick midguts and suggested a role for the JAK/STAT pathway in the inhibition of apoptosis in tick cells infected with A. phagocytophilum. Nevertheless, tick

  5. Host, habitat and climate preferences of Ixodes angustus (Acari: Ixodidae) and infection with Borrelia burgdorferi and Anaplasma phagocytophilum in California, USA.

    PubMed

    Stephenson, Nicole; Wong, Johnny; Foley, Janet

    2016-10-01

    The Holarctic tick Ixodes angustus is a competent vector for Borrelia burgdorferi, the etiologic agent of Lyme disease, and possibly Anaplasma phagocytophilum, the etiologic agent of granulocytic anaplasmosis, as well. From 2005 to 2013, we collected host-feeding I. angustus individuals from live-trapped small mammals and by flagging vegetation from 12 study sites in northern and central California, and tested for B. burgdorferi sensu lato, A. phagocytophilum, and Rickettsia spp. DNA by real-time PCR. Among 261 I. angustus collected (259 from hosts and two by flagging), the most common hosts were tree squirrels (20 % of ticks) and chipmunks (37 %). The PCR-prevalence for A. phagocytophilum and B. burgdorferi in ticks was 2 % and zero, respectively. The minimum infection prevalence on pooled DNA samples was 10 % for Rickettsia spp. DNA sequencing of the ompA gene identified this rickettsia as Candidatus Rickettsia angustus, a putative endosymbiont. A zero-inflated negative binomial mixed effects model was used to evaluate geographical and climatological predictors of I. angustus burden. When host species within study site and season within year were included in the model as nested random effects, all significant variables revealed that I. angustus burden increased as temperature decreased. Together with published data, these findings suggest that I. angustus is a host generalist, has a broad geographic distribution, is more abundant in areas with lower temperature within it's range, and is rarely infected with the pathogens A. phagocytophilum and B. burgdorferi.

  6. Prevalence Rates of Borrelia burgdorferi (Spirochaetales: Spirochaetaceae), Anaplasma phagocytophilum (Rickettsiales: Anaplasmataceae), and Babesia microti (Piroplasmida: Babesiidae) in Host-Seeking Ixodes scapularis (Acari: Ixodidae) from Pennsylvania.

    PubMed

    Hutchinson, M L; Strohecker, M D; Simmons, T W; Kyle, A D; Helwig, M W

    2015-07-01

    The etiological agents responsible for Lyme disease (Borrelia burgdorferi), human granulocytic anaplasmosis (Anaplasma phagocytophilum), and babesiosis (Babesia microti) are primarily transmitted by the blacklegged tick, Ixodes scapularis Say. Despite Pennsylvania having in recent years reported the highest number of Lyme disease cases in the United States, relatively little is known regarding the geographic distribution of the vector and its pathogens in the state. Previous attempts at climate-based predictive modeling of I. scapularis occurrence have not coincided with the high human incidence rates in parts of the state. To elucidate the distribution and pathogen infection rates of I. scapularis, we collected and tested 1,855 adult ticks statewide from 2012 to 2014. The presence of I. scapularis and B. burgdorferi was confirmed from all 67 Pennsylvania counties. Analyses were performed on 1,363 ticks collected in the fall of 2013 to avoid temporal bias across years. Infection rates were highest for B. burgdorferi (47.4%), followed by Ba. microti (3.5%) and A. phagocytophilum (3.3%). Coinfections included B. burgdorferi+Ba. microti (2.0%), B. burgdorferi+A. phagocytophilum (1.5%) and one tick positive for A. phagocytophilum+Ba. microti. Infection rates for B. burgdorferi were lower in the western region of the state. Our findings substantiate that Lyme disease risk is high throughout Pennsylvania.

  7. Serological detection of Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato and Ehrlichia canis antibodies and Dirofilaria immitis antigen in a countrywide survey in dogs in Poland.

    PubMed

    Krämer, Friederike; Schaper, Roland; Schunack, Bettina; Połozowski, Andrzej; Piekarska, Jolanta; Szwedko, Aleksandra; Jodies, Robert; Kowalska, Dagmara; Schüpbach, Dörte; Pantchev, Nikola

    2014-09-01

    Canine vector-borne diseases (CVBDs) have increasingly become a focus of attention in the past few years. Nevertheless, in many parts of Europe information on their occurrence is still scarce. In a large study in Poland 3,094 serum samples taken from dogs throughout all 16 Polish provinces were tested using a commercial kit for the detection of circulating antibodies against Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato and Ehrlichia canis and of Dirofilaria immitis antigen. A total of 12.31% (381/3,094; 95% confidence interval [CI]: 11.18-13.52%) and 3.75% (116/3,094; 95% CI: 3.11-4.48%) of the dogs were positive for A. phagocytophilum and B. burgdorferi s.l. antibodies, respectively. Furthermore, 0.26% (8/3,094; 95% CI: 0.11-0.51%) were positive for E. canis antibodies and 0.16% (5/3,094; 95% CI: 0.05-0.38%) for D. immitis antigen. The highest percentages of A. phagocytophilum-positive dogs were noted in Lesser Poland, Silesia and Łódź Provinces. For B. burgdorferi s.l., the highest prevalence was recorded in Łódź Province. Co-infections with A. phagocytophilum and B. burgdorferi s.l. were recorded in 1.71% of all examined dogs (53/3,094; 95% CI: 1.29-2.23%). One dog even had a triple infection, testing positive for E. canis too. Both A. phagocytophilum and B. burgdorferi s.l. have previously been reported in Poland and were confirmed in the present study by positive samples from all 16 provinces. Concerning E. canis and D. immitis travel history or importation cannot be excluded as factors which may have determined the occurrence of these pathogens in the relevant animals. Practitioners in Poland should be aware of the above mentioned CVBDs and of prophylactic measures to protect dogs and their owners.

  8. Prevalence and Distribution of Human and Tick Infections with the Ehrlichia muris-Like Agent and Anaplasma phagocytophilum in Wisconsin, 2009-2015.

    PubMed

    Murphy, Darby S; Lee, Xia; Larson, Scott R; Johnson, Diep K Hoang; Loo, Theoren; Paskewitz, Susan M

    2017-04-01

    Ehrlichiosis and anaplasmosis are important emerging tickborne zoonoses that affect both humans and animals. Knowledge of the geographic distribution and prevalence of Ehrlichia spp. and Anaplasma phagocytophilum in Wisconsin is important information as a baseline for future comparisons. Reported human cases between 2009 and 2015 were identified using the Wisconsin Electronic Disease Surveillance System (WEDSS) and mapped by county of residence. Vector surveillance was established using ticks collected from animals by partners, including veterinary medical clinics, domestic animal shelters, and wildlife rehabilitation centers from 40 Wisconsin counties. A total of 1835 Ixodes scapularis tick specimens (larvae, nymphs, and adults) were collected from 18 different domestic and wildlife species from July 2011 to November 2015. An additional 1136 nymphs were collected by drag sampling at 23 locations in 19 counties in 2015. A real-time PCR assay that detects and distinguishes several Ehrlichia species, including a pathogenic Ehrlichia muris-like agent (EMLA), and A. phagocytophilum was performed on adult and nymphal ticks. A total of 757 I. scapularis ticks (predominately adults) were tested from animal collections, with 67 (8.9%) individuals positive for A. phagocytophilum and 22 (2.9%) positive for EMLA DNA. Of the 1150 questing nymphs, 62 (5.4%) were positive for A. phagocytophilum and 10 (0.9%) were positive for EMLA DNA. Specimens of I. scapularis that were positive for A. phagocytophilum were found in 27 of the 33 counties surveyed. Specimens that were positive for EMLA were less common and were found in nine counties. This study provides the first statewide survey of I. scapularis ticks for these pathogens and indicates that the risk of human exposure is widely distributed.

  9. A new multiple-locus variable-number tandem repeat analysis reveals different clusters for Anaplasma phagocytophilum circulating in domestic and wild ruminants.

    PubMed

    Dugat, Thibaud; Chastagner, Amélie; Lagrée, Anne-Claire; Petit, Elisabeth; Durand, Benoît; Thierry, Simon; Corbière, Fabien; Verheyden, Hélène; Chabanne, Luc; Bailly, Xavier; Leblond, Agnès; Vourc'h, Gwenaël; Boulouis, Henri-Jean; Maillard, Renaud; Haddad, Nadia

    2014-09-16

    Anaplasma phagocytophilum is a tick-borne intragranulocytic alpha-proteobacterium. It is the causative agent of tick-borne fever in ruminants, and of human granulocytic anaplasmosis in humans, two diseases which are becoming increasingly recognized in Europe and the USA. However, while several molecular typing tools have been developed over the last years, few of them are appropriate for in-depth exploration of the epidemiological cycle of this bacterium. Therefore we have developed a Multiple-Locus Variable number tandem repeat (VNTR) Analysis typing technique for A. phagocytophilum. Five VNTRs were selected based on the HZ human-derived strain genome, and were tested on the Webster human-derived strain and on 123 DNA samples: 67 from cattle, 7 from sheep, 15 from roe deer, 4 from red deer, 1 from a reindeer, 2 from horses, 1 from a dog, and 26 from ticks. From these samples, we obtained 84 different profiles, with a diversity index of 0.96 (0.99 for vertebrate samples, i.e. without tick samples). Our technique confirmed that A. phagocytophilum from roe deer or domestic ruminants belong to two different clusters, while A. phagocytophilum from red deer and domestic ruminants locate within the same cluster, questioning the respective roles of roe vs red deer as reservoir hosts for domestic ruminant strains in Europe. As expected, greater diversity was obtained between rather than within cattle herds. Our technique has great potential to provide detailed information on A. phagocytophilum isolates, improving both epidemiological and phylogenic investigations, thereby helping in the development of relevant prevention and control measures.

  10. Serological survey of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, and Ehrlichia canis infections in rural and urban dogs in Central Italy.

    PubMed

    Ebani, Valentina Virginia; Bertelloni, Fabrizio; Torracca, Beatrice; Cerri, Domenico

    2014-01-01

    Borrelia burgdorferi sensu lato (s.l.) and Anaplasma phagocytophilum are well known zoonotic pathogens, whereas Ehrlichia canis is usually considered to be of veterinary concern, although on the basis of recent reports it also seems to be able to infect humans. The aim of the study was to determine the seroprevalence of B. burgdorferi s.l., A. phagocytophilum and E. canis in an Italian canine population, and to verify if there are differences between dogs living in urban areas and those from a rural environment. Blood sera of 1,965 dogs, 1,235 from cities and 730 from rural areas, were tested by indirect immunofluorescent assay (IFAT). The overall seroprevalence was highest for E. canis (7.07%), followed by A. phagocytophilum (4.68%), and B. burgdorferi s.l. (1.47%). Rural dogs showed the highest seroprevalence to B. burgdorferi s.l. and A. phagocytophilum. No significant differences were observed between rural and urban E. canis-positive dogs. A low percentage (1.32%) of dogs with dual seropositivity was detected, and no triple positive reactions were observed. No significant differences were detected in the seroprevalence of the three agents in relationship to the age and gender of the dogs. Seroprevalence in the five years considered were not statistically different, except for the lowest rate for E. canis observed in 2012. The results confirm the presence of B. burgdorferi s.l., A. phagocytophilum and E. canis in Italian dogs in both urban and rural areas. Monitoring pet dogs, which share the same environment with their owners, is useful for identifying the presence of tick-borne disease agents of both veterinary and public health significance.

  11. Anaplasma phagocytophilum APH0032 Is Exposed on the Cytosolic Face of the Pathogen-Occupied Vacuole and Co-opts Host Cell SUMOylation

    PubMed Central

    Oki, Aminat T.; Huang, Bernice; Beyer, Andrea R.; May, Levi J.; Truchan, Hilary K.; Walker, Naomi J.; Galloway, Nathan L.; Borjesson, Dori L.; Carlyon, Jason A.

    2016-01-01

    Anaplasma phagocytophilum, a member of the family Anaplasmataceae and the obligate intracellular bacterium that causes granulocytic anaplasmosis, resides in a host cell-derived vacuole. Bacterial proteins that localize to the A. phagocytophilum-occupied vacuole membrane (AVM) are critical host-pathogen interfaces. Of the few bacterial AVM proteins that have been identified, the domains responsible for AVM localization and the host cell pathways that they co-opt are poorly defined. APH0032 is an effector that is expressed and localizes to the AVM late during the infection cycle. Herein, the APH0032 domain that is essential for associating with host cell membranes was mapped. Immunofluorescent labeling of infected cells that had been differentially permeabilized confirmed that APH0032 is exposed on the AVM's cytosolic face, signifying its potential to interface with host cell processes. SUMOylation is the covalent attachment of a member of the small ubiquitin-like modifier (SUMO) family of proteins to lysines in target substrates. Previous work from our laboratory determined that SUMOylation is important for A. phagocytophilum survival and that SUMOylated proteins decorate the AVM. Algorithmic prediction analyses identified APH0032 as a candidate for SUMOylation. Endogenous APH0032 was precipitated from infected cells using a SUMO affinity matrix, confirming that the effector co-opts SUMOylation during infection. APH0032 pronouncedly colocalized with SUMO1, but not SUMO2/3 moieties on the AVM. Ectopic expression of APH0032 in A. phagocytophilum infected host cells significantly boosted the bacterial load. This study delineates the first domain of any Anaplasmataceae protein that is essential for associating with the pathogen-occupied vacuole membrane, demonstrates the importance of APH0032 to infection, and identifies it as the second A. phagocytophilum effector that co-opts SUMOylation, thus underscoring the relevance of this post-translational modification to

  12. Anaplasma phagocytophilum increases the levels of histone modifying enzymes to inhibit cell apoptosis and facilitate pathogen infection in the tick vector Ixodes scapularis

    PubMed Central

    Cabezas-Cruz, Alejandro; Alberdi, Pilar; Ayllón, Nieves; Valdés, James J.; Pierce, Raymond; Villar, Margarita; de la Fuente, José

    2016-01-01

    ABSTRACT Epigenetic mechanisms have not been characterized in ticks despite their importance as vectors of human and animal diseases worldwide. The objective of this study was to characterize the histones and histone modifying enzymes (HMEs) of the tick vector Ixodes scapularis and their role during Anaplasma phagocytophilum infection. We first identified 5 histones and 34 HMEs in I. scapularis in comparison with similar proteins in model organisms. Then, we used transcriptomic and proteomic data to analyze the mRNA and protein levels of I. scapularis histones and HMEs in response to A. phagocytophilum infection of tick tissues and cultured cells. Finally, selected HMEs were functionally characterized by pharmacological studies in cultured tick cells. The results suggest that A. phagocytophilum manipulates tick cell epigenetics to increase I. scapularis p300/CBP, histone deacetylase, and Sirtuin levels, resulting in an inhibition of cell apoptosis that in turn facilitates pathogen infection and multiplication. These results also suggest that a compensatory mechanism might exist by which A. phagocytophilum manipulates tick HMEs to regulate transcription and apoptosis in a tissue-specific manner to facilitate infection, but preserving tick fitness to guarantee survival of both pathogens and ticks. Our study also indicates that the pathogen manipulates arthropod and vertebrate cell epigenetics in similar ways to inhibit the host response to infection. Epigenetic regulation of tick biological processes is an essential element of the infection by A. phagocytophilum and the study of the mechanisms and principal actors involved is likely to provide clues for the development of anti-tick drugs and vaccines. PMID:27019326

  13. Are Apodemus spp. mice and Myodes glareolus reservoirs for Borrelia miyamotoi, Candidatus Neoehrlichia mikurensis, Rickettsia helvetica, R. monacensis and Anaplasma phagocytophilum?

    PubMed

    Burri, C; Schumann, O; Schumann, C; Gern, L

    2014-04-01

    In Europe, in addition to Borrelia burgdorferi sensu lato and tick-borne encephalitis (TBE) virus, other zoonotic pathogens, like B. miyamotoi, a species related to the relapsing fever spirochaetes, Candidatus Neoehrlichia mikurensis (N. mikurensis), Rickettsia helvetica, Rickettsia monacensis, and Anaplasma phagocytophilum have been reported in the ixodid tick Ixodes ricinus. No study was conducted to identify reservoir hosts for these pathogens. Here, we investigated the role played by wild rodents in the natural transmission cycle of B. miyamotoi, N. mikurensis, R. helvetica, R. monacensis, and A. phagocytophilum in Switzerland. In 2011 and 2012, small mammals were captured in an area where these pathogens occur in questing ticks. Ixodes ricinus ticks infesting captured small mammals were analysed after their moult by PCR followed by reverse line blot to detect the different pathogens. Xenodiagnostic larvae were used to evaluate the role of rodents as reservoirs and analysed after their moult. Most of the 108 captured rodents (95.4%) were infested by I. ricinus ticks; 4.9%, 3.9%, 24.0%, and 0% of the rodents were infested by Borrelia, N. mikurensis, Rickettsia spp., and A. phagocytophilum-infected larvae, respectively. Borrelia afzelii, B. miyamotoi, N. mikurensis, Rickettsia spp., and A. phagocytophilum were detected in 2.8%, 0.17%, 2.6%, 6.8%, and 0% of the ticks attached to rodents, respectively. Borrelia afzelii was transmitted by 4 rodents to 41.2% of the xenodiagnostic ticks, B. miyamotoi by 3 rodents to 23.8%, and N. mikurensis was transmitted by 6 rodents to 41.0% of the xenodiagnostic ticks. None of the tested rodent transmitted Rickettsia spp. or A. phagocytophilum to I. ricinus xenodiagnostic larvae. This study showed that rodents are reservoir hosts for B. miyamotoi and N. mikurensis in Europe.

  14. Seroprevalence of Borrelia burgdorferi, Anaplasma phagocytophilum, and Francisella tularensis Infections in Belgium: Results of Three Population-Based Samples.

    PubMed

    De Keukeleire, Mathilde; Vanwambeke, Sophie O; Cochez, Christel; Heyman, Paul; Fretin, David; Deneys, Véronique; Luyasu, Victor; Kabamba, Benoît; Robert, Annie

    2017-02-01

    To estimate the seroprevalence of Borrelia burgdorferi (Bb), Anaplasma phagocytophilum (Ap), and Francisella tularensis (Ft) in Belgium, we tested sera from three population-based samples in which exposure to pathogen is assumed to vary: 148 samples from workers professionally exposed, 209 samples from rural blood donors, and 193 samples from urban blood donors. Sera were tested using ELISA or the immunofluorescence assay test. The seroprevalence of Bb was 5.4% in workers professionally exposed, 2.9% in rural blood donors, and 2.6% in urban blood donors, which is similar to other studies. The fraction of negative results decreases significantly from urban blood donors and rural blood donors to workers. Regarding the seroprevalence of Ap, the cutoff titer of 1:64 recommended by the manufacturer may be set too low and produces artificially high seroprevalence rates. Using a cutoff titer of 1:128, the seroprevalence of Ap was estimated at 8.1% for workers professionally exposed, 6.2% for rural blood donors, and 5.7% for urban blood donors. Tularemia sera confirmed the presence of the pathogen in Belgium at 2.0% for workers and 0.5% for rural and urban blood donors. Our study is one of the few providing an estimation of the seroprevalences of Bb, Ap, and Ft in three different populations in Belgium, filling the gap in seroprevalence data among those groups. Our findings provide evidence that the entire Belgian population is exposed to Bb, Ap, and Ft infections, but a higher exposure is noticed for professionals at risk. Education on the risk factors for tick bites and preventive measures for both professionals exposed and the general population is needed.

  15. Emergence of tick-borne pathogens (Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Ricketsia raoultii and Babesia microti) in the Kyiv urban parks, Ukraine.

    PubMed

    Didyk, Yuliya M; Blaňárová, Lucia; Pogrebnyak, Svyatoslav; Akimov, Igor; Peťko, Branislav; Víchová, Bronislava

    2017-02-01

    To date, only limited data about the presence of ticks and circulation of tick-borne pathogens in urban parks of Kyiv in northern Ukraine are available. In total, 767 ticks (696 Ixodes ricinus and 69 Dermacentor reticulatus) collected in seven urban parks and one suburban oak wood park in Kyiv were individually analyzed by the PCR assays. Tick-borne pathogens, namely spirochetes from Borrelia burgdorferi sensu lato complex, Anaplasma phagocytophilum, and Babesia microti, were detected in 11.1% of tested I. ricinus ticks. In total, 4% of I. ricinus ticks tested positive for the presence of B. burdorferi s.l. (Borrelia afzelii and Borrelia garinii), 5.2% for A. phagocytophilum, and Ba. microti was confirmed in 1.9% of examined ticks. Mixed infections were recorded in four DNA samples, representing the prevalence of 0.6%. One female and two I. ricinus nymphs were simultaneously infected with B. afzelii and A. phagocytophilum, and one female carried B. afzelii and Ba. microti. In addition, 10.1% of D. reticulatus ticks tested positive for Rickettsia raoultii. Identification of infectious agents and their diversity, assessment of the relative epidemiological importance and determination of the prevalence in questing ticks from central parts of the cities are crucial steps towards the tick-borne diseases surveillance in urban environment.

  16. Serologic and Molecular Prevalence of Rickettsia helvetica and Anaplasma phagocytophilum in Wild Cervids and Domestic Mammals in the Central Parts of Sweden.

    PubMed

    Elfving, Karin; Malmsten, Jonas; Dalin, Anne-Marie; Nilsson, Kenneth

    2015-09-01

    Both Rickettsia helvetica and Anaplasma phagocytophilum are common in Ixodes ricinus ticks in Sweden. Knowledge is limited regarding different animal species' competence to act as reservoirs for these organism. For this reason, blood samples were collected from wild cervids (roe deer, moose) and domestic mammals (horse, cat, dog) in central Sweden, and sera were tested using immunofluorescence assay to detect antibodies against spotted fever rickettsiae using Rickettsia helvetica as antigen. Sera with a titer ≥1:64 were considered as positive, and 23.1% (104/450) of the animals scored positive. The prevalence of seropositivity was 21.5% (23/107) in roe deer, 23.3% (21/90) in moose, 36.5% (23/63) in horses, 22.1% (19/90) in cats, and 17.0% (17/100) in dogs. PCR analysis of 113 spleen samples from moose and sheep from the corresponding areas were all negative for rickettsial DNA. In roe deer, 85% (91/107) also tested seropositive for A. phagocytophilum with a titer cutoff of 1:128. The findings indicate that the surveyed animal species are commonly exposed to rickettsiae and roe deer also to A. phagocytophilum.

  17. Molecular detection of co-infections with Anaplasma phagocytophilum and/or Babesia canis canis in Dirofilaria-positive dogs from Slovakia.

    PubMed

    Víchová, Bronislava; Miterpáková, Martina; Iglódyová, Adriana

    2014-06-16

    Recently, several arthropod-borne infections have been introduced into previously non-endemic regions in Europe as the result of various global changes. At the same time, endemic regions are expanding and the risk of co-infections is rising, due to climate change that allows vectors to move and spread infectious diseases into new areas. The aim of the current study was to confirm simultaneous infections with Anaplasma phagocytophilum and/or Babesia canis canis in Dirofilaria-infected dogs from Slovakia, central Europe. Genomic DNA was isolated from 366 blood samples of microfilaraemic dogs without clinical signs of infection. Samples were further screened for the presence of canine tick-borne pathogens using PCR and sequencing. This survey revealed co-infection with four arthropod-borne pathogens, in particular, Dirofilaria repens, Dirofilaria immitis, A. phagocytophilum, and B. canis canis. While D. repens, responsible for canine subcutaneous dirofilariosis, is scattered through the whole territory of the country, D. immitis occurs only in endemic areas of southeastern and southwestern Slovakia in mixed infection with D. repens. Co-infection with A. phagocytophilum was reported in 3.27% of the dogs positive for D. repens; mixed infection with D. repens and B. canis canis was detected in 3.55% of the tested blood samples. Eastern Slovak Lowland represents a natural focus of B. canis canis and is a highly endemic area for canine dirofilariosis. The presence of triple infection with D. repens, A. phagocytophilum, and B. canis canis was detected in one dog originating from the eastern lowland region of Slovakia. This study highlights the importance of co-infected, clinically healthy dogs in the spreading of several different arthropod-borne pathogens and the necessity for detailed epidemiological surveys, especially in newly infested areas. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Host specificity, pathogen exposure, and superinfections impact the distribution of Anaplasma phagocytophilum genotypes in ticks, roe deer, and livestock in a fragmented agricultural landscape.

    PubMed

    Chastagner, Amélie; Pion, Angélique; Verheyden, Hélène; Lourtet, Bruno; Cargnelutti, Bruno; Picot, Denis; Poux, Valérie; Bard, Émilie; Plantard, Olivier; McCoy, Karen D; Leblond, Agnes; Vourc'h, Gwenaël; Bailly, Xavier

    2017-08-12

    Anaplasma phagocytophilum is a bacterial pathogen mainly transmitted by Ixodes ricinus ticks in Europe. It infects wild mammals, livestock, and, occasionally, humans. Roe deer are considered to be the major reservoir, but the genotypes they carry differ from those that are found in livestock and humans. Here, we investigated whether roe deer were the main source of the A. phagocytophilum genotypes circulating in questing I. ricinus nymphs in a fragmented agricultural landscape in France. First, we assessed pathogen prevalence in 1837 I. ricinus nymphs (sampled along georeferenced transects) and 79 roe deer. Prevalence was dramatically different between ticks and roe deer: 1.9% versus 76%, respectively. Second, using high-throughput amplicon sequencing, we characterized the diversity of the A. phagocytophilum genotypes found in 22 infected ticks and 60 infected roe deer; the aim was to determine the frequency of co-infections. Only 22.7% of infected ticks carried genotypes associated with roe deer. This finding fits with others suggesting that cattle density is the major factor explaining infected tick density. To explore epidemiological scenarios capable of explaining these patterns, we constructed compartmental models that focused on how A. phagocytophilum exposure and infection dynamics affected pathogen prevalence in roe deer. At the exposure levels predicted by the results of this study and the literature, the high prevalence in roe deer was only seen in the model in which superinfections could occur during all infection phases and when the probability of infection post exposure was above 0.43. We then interpreted these results from the perspective of livestock and human health. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Detection and quantification of Anaplasma phagocytophilum and Babesia spp. in Ixodes ricinus ticks from urban and rural environment, northern Poland, by real-time polymerase chain reaction.

    PubMed

    Stańczak, Joanna; Cieniuch, Stella; Lass, Anna; Biernat, Beata; Racewicz, Maria

    2015-05-01

    Anaplasma phagocytophilum and Babesia spp. are emerging tick-borne pathogens which can threaten human health. A duplex real-time PCR and qPCRs with primers and probes targeting 97 and 116 bp fragments of 16S rRNA and 18S rRNA genes, respectively, were used for qualitative and quantitative detection of both pathogens in Ixodes ricinus ticks. Altogether 1875 ticks (1084 adults and 791 nymphs) were collected from rural and urban habitats in northern Poland. Of them, at least 0.9% were found to be infected with A. phagocytophilum while 2.5% with Babesia spp. A comparison of the infection rates by the tick stage, the type of area, the collection site, habitats of different tick density and by the month of collection was done. The prevalence of pathogens was significantly lower in nymphs than in adult ticks (p = 0.02) and in rural areas than in urban areas (p = 0.007). Four different 16S rRNA gene variants of A. phagocytophilum were determine, however none of them showed 100% identity with compared sequences isolated from human patients. The dominant Babesia species was B. venatorum. Results of qPCRs with circular and linearized forms of plasmids used as the standards showed significant difference in the pathogen loads (p = 0.001). The copy numbers of A. phagocytophilum and Babesia spp. estimated from the linear plasmids were 28.7 and 5.1 times lower, respectively, when compared with their circular forms, and were accepted as more reliable. The average number of copies of 16S rRNA gene of A. phagocytophilum in the positive I. ricinus samples were 3.39 × 10(5) ± 6.09 × 10(5). The mean copy number of 18S rRNA gene of Babesia spp. was ~2.55 × 10(5) ± 1.04 × 10(6). We confirmed the presence of A. phagocytophilum and Babesia spp. in I. ricinus in both rural and urban environments. The determined low infection rates suggests, however, that the risk for local population and tourists to acquire infection is also low. Moreover, we confirmed recent findings that serious

  20. Are patients with erythema migrans who have leukopenia and/or thrombocytopenia coinfected with Anaplasma phagocytophilum or tick-borne encephalitis virus?

    PubMed

    Strle, Franc; Bogovič, Petra; Cimperman, Jože; Maraspin, Vera; Ogrinc, Katarina; Rojko, Tereza; Stupica, Daša; Lusa, Lara; Avšič-Županc, Tatjana; Smrdel, Katja Strašek; Jelovšek, Mateja; Lotrič-Furlan, Stanka

    2014-01-01

    Lyme borreliosis (LB), tick-borne encephalitis (TBE) and human granulocytic anaplasmosis (HGA) are endemic in central part of Slovenia. We tested the hypothesis that patients with erythema migrans (EM) from this region, who have leukopenia and/or thrombocytopenia (typical findings in HGA and in the initial phase of TBE but not in patients with LB) are coinfected with Anaplasma phagocytophilum and/or with TBE virus, i.e. that cytopenia is a result of concomitant HGA or the initial phase of TBE. Comparison of clinical and laboratory findings for 67 patients with EM who disclosed leukopenia/thrombocytopenia with the corresponding results in sex- and age-matched patients with EM and normal blood cell counts revealed no differences. In addition, patients with typical EM and leukopenia and/or thrombocytopenia tested negative for the presence of IgM and IgG antibodies to TBE virus by ELISA as well as for the presence of specific IgG antibodies to A. phagocytophilum antigens by IFA in acute and convalescent serum samples. Thus, none of 67 patients (95% CI: 0 to 5.3%) with typical EM (the presence of this skin lesion attests for early Lyme borreliosis and is the evidence for a recent tick bite) was found to be coinfected with A. phagocytophilum or had a recent primary infection with TBE virus. The findings in the present study indicate that in Slovenia, and probably in other European countries endemic for LB, TBE and HGA, patients with early LB are rarely coinfected with the other tick-transmitted agents.

  1. Genome-Wide Anaplasma phagocytophilum AnkA-DNA Interactions Are Enriched in Intergenic Regions and Gene Promoters and Correlate with Infection-Induced Differential Gene Expression

    PubMed Central

    Dumler, J. Stephen; Sinclair, Sara H.; Pappas-Brown, Valeria; Shetty, Amol C.

    2016-01-01

    Anaplasma phagocytophilum, an obligate intracellular prokaryote, infects neutrophils, and alters cardinal functions via reprogrammed transcription. Large contiguous regions of neutrophil chromosomes are differentially expressed during infection. Secreted A. phagocytophilum effector AnkA transits into the neutrophil or granulocyte nucleus to complex with DNA in heterochromatin across all chromosomes. AnkA binds to gene promoters to dampen cis-transcription and also has features of matrix attachment region (MAR)-binding proteins that regulate three-dimensional chromatin architecture and coordinate transcriptional programs encoded in topologically-associated chromatin domains. We hypothesize that identification of additional AnkA binding sites will better delineate how A. phagocytophilum infection results in reprogramming of the neutrophil genome. Using AnkA-binding ChIP-seq, we showed that AnkA binds broadly throughout all chromosomes in a reproducible pattern, especially at: (i) intergenic regions predicted to be MARs; (ii) within predicted lamina-associated domains; and (iii) at promoters ≤ 3000 bp upstream of transcriptional start sites. These findings provide genome-wide support for AnkA as a regulator of cis-gene transcription. Moreover, the dominant mark of AnkA in distal intergenic regions known to be AT-enriched, coupled with frequent enrichment in the nuclear lamina, provides strong support for its role as a MAR-binding protein and genome “re-organizer.” AnkA must be considered a prime candidate to promote neutrophil reprogramming and subsequent functional changes that belie improved microbial fitness and pathogenicity. PMID:27703927

  2. Role of migratory birds in introduction and range expansion of Ixodes scapularis ticks and of Borrelia burgdorferi and Anaplasma phagocytophilum in Canada.

    PubMed

    Ogden, N H; Lindsay, L R; Hanincová, K; Barker, I K; Bigras-Poulin, M; Charron, D F; Heagy, A; Francis, C M; O'Callaghan, C J; Schwartz, I; Thompson, R A

    2008-03-01

    During the spring in 2005 and 2006, 39,095 northward-migrating land birds were captured at 12 bird observatories in eastern Canada to investigate the role of migratory birds in northward range expansion of Lyme borreliosis, human granulocytic anaplasmosis, and their tick vector, Ixodes scapularis. The prevalence of birds carrying I. scapularis ticks (mostly nymphs) was 0.35% (95% confidence interval [CI] = 0.30 to 0.42), but a nested study by experienced observers suggested a more realistic infestation prevalence of 2.2% (95% CI = 1.18 to 3.73). The mean infestation intensity was 1.66 per bird. Overall, 15.4% of I. scapularis nymphs (95% CI = 10.7 to 20.9) were PCR positive for Borrelia burgdorferi, but only 8% (95% CI = 3.8 to 15.1) were positive when excluding nymphs collected at Long Point, Ontario, where B. burgdorferi is endemic. A wide range of ospC and rrs-rrl intergenic spacer alleles of B. burgdorferi were identified in infected ticks, including those associated with disseminated Lyme disease and alleles that are rare in the northeastern United States. Overall, 1.4[corrected]% (95% CI = 0.3 [corrected] to 0.41) of I. scapularis nymphs were PCR positive for Anaplasma phagocytophilum. We estimate that migratory birds disperse 50 million to 175 million I. scapularis ticks across Canada each spring, implicating migratory birds as possibly significant in I. scapularis range expansion in Canada. However, infrequent larvae and the low infection prevalence in ticks carried by the birds raise questions as to how B. burgdorferi and A. phagocytophilum become endemic in any tick populations established by bird-transported ticks.

  3. Role of Migratory Birds in Introduction and Range Expansion of Ixodes scapularis Ticks and of Borrelia burgdorferi and Anaplasma phagocytophilum in Canada▿

    PubMed Central

    Ogden, N. H.; Lindsay, L. R.; Hanincová, K.; Barker, I. K.; Bigras-Poulin, M.; Charron, D. F.; Heagy, A.; Francis, C. M.; O'Callaghan, C. J.; Schwartz, I.; Thompson, R. A.

    2008-01-01

    During the spring in 2005 and 2006, 39,095 northward-migrating land birds were captured at 12 bird observatories in eastern Canada to investigate the role of migratory birds in northward range expansion of Lyme borreliosis, human granulocytic anaplasmosis, and their tick vector, Ixodes scapularis. The prevalence of birds carrying I. scapularis ticks (mostly nymphs) was 0.35% (95% confidence interval [CI] = 0.30 to 0.42), but a nested study by experienced observers suggested a more realistic infestation prevalence of 2.2% (95% CI = 1.18 to 3.73). The mean infestation intensity was 1.66 per bird. Overall, 15.4% of I. scapularis nymphs (95% CI = 10.7 to 20.9) were PCR positive for Borrelia burgdorferi, but only 8% (95% CI = 3.8 to 15.1) were positive when excluding nymphs collected at Long Point, Ontario, where B. burgdorferi is endemic. A wide range of ospC and rrs-rrl intergenic spacer alleles of B. burgdorferi were identified in infected ticks, including those associated with disseminated Lyme disease and alleles that are rare in the northeastern United States. Overall, 0.4% (95% CI = 0.03 to 0.41) of I. scapularis nymphs were PCR positive for Anaplasma phagocytophilum. We estimate that migratory birds disperse 50 million to 175 million I. scapularis ticks across Canada each spring, implicating migratory birds as possibly significant in I. scapularis range expansion in Canada. However, infrequent larvae and the low infection prevalence in ticks carried by the birds raise questions as to how B. burgdorferi and A. phagocytophilum become endemic in any tick populations established by bird-transported ticks. PMID:18245258

  4. Occurrence of ticks and prevalence of Anaplasma phagocytophilum and Borrelia burgdorferi s.l. in three types of urban biotopes: forests, parks and cemeteries.

    PubMed

    Hornok, Sándor; Meli, Marina L; Gönczi, Enikő; Halász, Edina; Takács, Nóra; Farkas, Róbert; Hofmann-Lehmann, Regina

    2014-10-01

    The aim of the present study was to compare different urban biotopes for the occurrence of ixodid tick species, for the population density of Ixodes ricinus and for the prevalence rates of two emerging, zoonotic pathogens. Altogether 2455 ticks were collected from the vegetation on 30 places (forests, parks, cemeteries) of Budapest, Hungary. I. ricinus and Haemaphysalis concinna were collected in all three biotope types, but Dermacentor reticulatus only in parks and forests, and D. marginatus only in a forest. Highest population density of I. ricinus was observed in neglected parts of cemeteries. In females of this tick species the prevalence rates of Anaplasma phagocytophilum and Borrelia burgdorferi s.l. were significantly lower in cemeteries, than in parks or forests. In conclusion, risks associated with the presence of ticks and tick-borne pathogens may be high in a city, but this depends on biotope types, due to habitat-related differences in the vegetation, as well as in the availability of tick hosts and pathogen reservoirs.

  5. Anaplasma species of veterinary importance in Japan.

    PubMed

    Ybañez, Adrian Patalinghug; Inokuma, Hisashi

    2016-11-01

    Anaplasma species of the family Anaplasmataceae, order Rickettsiales are tick-borne organisms that can cause disease in animals and humans. In Japan, all recognized species of Anaplasma (except for Anaplasma ovis) and a potentially novel Anaplasma sp. closely related to Anaplasma phagocytophilum have been reported. Most of these detected tick-borne pathogens are believed to be lowly pathogenic in animals in Japan although the zoonotic A. phagocytophilum has recently been reported to cause clinical signs in a dog and in humans. This review documents the studies and reports about Anaplasma spp. in Japan.

  6. Anaplasma species of veterinary importance in Japan

    PubMed Central

    Ybañez, Adrian Patalinghug; Inokuma, Hisashi

    2016-01-01

    Anaplasma species of the family Anaplasmataceae, order Rickettsiales are tick-borne organisms that can cause disease in animals and humans. In Japan, all recognized species of Anaplasma (except for Anaplasma ovis) and a potentially novel Anaplasma sp. closely related to Anaplasma phagocytophilum have been reported. Most of these detected tick-borne pathogens are believed to be lowly pathogenic in animals in Japan although the zoonotic A. phagocytophilum has recently been reported to cause clinical signs in a dog and in humans. This review documents the studies and reports about Anaplasma spp. in Japan. PMID:27956767

  7. Current Surveys of the Seroprevalence of Borrelia burgdorferi, Ehrlichia canis, Anaplasma phagocytophilum, Leishmania infantum, Babesia canis, Angiostrongylus vasorum and Dirofilaria immitis in Dogs in Bulgaria.

    PubMed

    Pantchev, Nikola; Schnyder, Manuela; Vrhovec, Majda Globokar; Schaper, Roland; Tsachev, Ilia

    2015-08-01

    Canine vector-borne diseases (CVBDs) have increasingly become a focus of interest in recent years. Some of the CVBDs are zoonotic and may therefore also represent a risk for the human population. Different factors are in discussion to explain the expansion of vectors and pathogens into formerly unaffected areas. Knowledge of the prevalence and distribution of CVBDs in Bulgaria is scant overall and most data rely on single case descriptions. The aim of the present study was to determine the seroprevalence of important CVBDs in 167 dogs from central-southern Bulgaria (Stara Zagora), with special emphasis on hitherto uninvestigated babesiosis and angiostrongylosis, on poorly investigated Lyme borreliosis and canine granulocytic anaplasmosis, and on the potentially zoonotic dirofilariosis and leishmaniosis. Relatively high prevalence rates were documented for anti-Babesia canis antibodies, Dirofilaria immitis antigen (16.2 %; 27/167 each), anti-Ehrlichia canis (21 %; 35/167) and anti-Anaplasma phagocytophilum antibodies (30.5 - 46.1 %; 51 - 77/167), while Borrelia burgdorferi seroprevalence was low (2.4 %; 4/167). All samples were negative for Leishmania infantum antibodies and Angiostrongylus vasorum antigen and antibodies. In total, 64.7 % (108/167) of the samples indicated infection or exposure to at least one agent and a high proportion of dual infections (39.8 %; 43/108) was demonstrated. Multiple infections with up to four different organisms were also detected. Our data underline the importance of CVBDs and especially of co-infections which could influence the clinical outcome in dogs.

  8. Evaluation of Doxycycline-Laden Oral Bait and Topical Fipronil Delivered in a Single Bait Box to Control Ixodes scapularis (Acari: Ixodidae) and Reduce Borrelia burgdorferi and Anaplasma phagocytophilum Infection in Small Mammal Reservoirs and Host-Seeking Ticks.

    PubMed

    Dolan, Marc C; Schulze, Terry L; Jordan, Robert A; Schulze, Christopher J; Ullmann, Amy J; Hojgaard, Andrias; Williams, Martin A; Piesman, Joseph

    2017-03-01

    A field trial was conducted on residential properties in a Lyme disease endemic area of New Jersey to determine the efficacy of Maxforce Tick Management System (TMS) bait boxes modified with doxycycline hyclate-laden bait to reduce the acarological risk of Lyme disease and the utility of galvanized steel shrouds to protect the bait boxes from squirrel depredation and ability to routinely service these devices. The strategy began with a 9-wk deployment against larvae followed by a 17-wk deployment against nymphs and larvae the second year. Passive application of fipronil reduced nymphal and larval tick burdens on small mammals by 76 and 77%, respectively, and nymphal tick abundance by 81% on treated properties. In addition, the percentage of infected small mammals recovered from intervention areas following treatment was reduced by 96% for Borrelia burgdorferi and 93% for Anaplasma phagocytophilum. Infection prevalence in host-seeking nymphal ticks for both B. burgdorferi and A. phagocytophilum were reduced by 93 and 61%, respectively. Results indicate that Maxforce TMS bait boxes fitted with doxycycline-impregnated bait is an effective means of reducing ticks and infection prevalence for B. burgdorferi and A. phagocytophilum in both rodent reservoirs and questing Ixodes scapularis Say ticks. The protective shroud allows the device to be routinely serviced and protect against squirrel depredation. Published by Oxford University Press on behalf of Entomological Society of America 2016. This work is written by US Government employees and is in the public domain in the US.

  9. A serological survey of tick-borne pathogens in dogs in North America and the Caribbean as assessed by Anaplasma phagocytophilum, A. platys, Ehrlichia canis, E. chaffeensis, E. ewingii, and Borrelia burgdorferi species-specific peptides

    PubMed Central

    Qurollo, Barbara A.; Chandrashekar, Ramaswamy; Hegarty, Barbara C.; Beall, Melissa J.; Stillman, Brett A.; Liu, Jiayou; Thatcher, Brendon; Pultorak, Elizabeth; Cerrito, Brian; Walsh, Mary; Breitschwerdt, Edward B.

    2014-01-01

    Introduction Tick-borne pathogens cause a spectrum of disease manifestations in both dogs and humans. Recognizing regional and temporal shifts in exposure are important as tick distributions change. To better delineate regional exposure to canine tick-borne pathogens, an expanded set of species-specific peptides were used to detect Anaplasma phagocytophilum (Aph), Anaplasma platys (Apl), Ehrlichia canis (Ec), Ehrlichia chaffeensis (Ech), Ehrlichia ewingii (Eew), and Borrelia burgdorferi (Bb) antibodies in canine serum. Methods Archived canine serum samples (n=6,582) collected during 2008–2010 and in 2012 from the US, Canada, and the Caribbean were retrospectively screened for antibodies against Ehrlichia and Anaplasma species-specific peptides. Overall, regional and temporal seroprevalence rates were determined. Results Overall Bb and Eew were the most seroprevalent pathogens. During 2008–2010, seroprevalence rates increased overall for Aph and Ech, and regionally, Bb and Aph seroprevalence rates increased in the South. Canada had unexpectedly high seroprevalence rates for Ec and Apl. The most common co-exposures were Eew+Ech, followed by Aph+Bb and Eew+Bb. Conclusions This study demonstrated significant shifts in canine vector-borne disease seroprevalence rates. The use of specific peptides facilitated improved geographic delineation of tick-borne pathogen distributions among dogs, which may enhance epidemiological surveillance of vector-borne pathogens shared by dogs and humans. PMID:25405006

  10. Analysis of main T-cell subsets and activated T supressor/cytotoxic cells in patients with Borrelia burgdorferi s. lato only infection and co-infections with Anaplasma phagocytophilum, Bartonella spp. and Babesia microti.

    PubMed

    Tomasiewicz, Krzysztof; Chmielewska-Badora, Jolanta; Zwolinski, Jacek; Murias-Brylowska, Elzbieta

    2016-01-01

    The study was designed to assess the role of some important immunologic factors with regards to both laboratory results and clinical symptoms in patients with confirmed Lyme disease. Additional examinations were carried out for co-infections with a number of tick-borne pathogens. The study group consisted of 54 patients with Lyme disease and a group of 21 healthy controls. Serology of co-infections with Anaplasma phagocytophilum, Bartonella spp. and Babesia microti was carrieed out in all patients. Blood samples were stained using the whole-blood lysis method and analyzed concurrently on a flow cytometer FACSCalibur. Directly conjugated anti-human monoclonal antibodies against CD3, CD4, CD8, CD16, CD56, HLA-DR and CD69 were used. No significant differences were observed with respect to thepretreatment level of CD4+ and CD8+ cells. In patients with symptoms relief and symptoms persistence, lower percentages of CD4+ and CD8+ cells were found, but with no statistical dependence. In the study group, both in patients with and without co-infections, pretreatment values of CD16+CD56+ cells did not differ significantly. In patients who did not respond to the treatment, the baseline percentage of NK cells was higher (P<0.01) than in group with clinical improvement, and lower after the treatment, whereas in patients with symptoms relief after the treatment there was an increase in the percentage of NK cells. Co-infections with Anaplasma phagocytophilum, Bartonella spp. and Babesia microti had no impact on T-cell percentages in Lyme disease patients. There was a lower baseline percentage of NK cells in patients not responding to antibiotic treatment.

  11. Infection and co-infection rates of Anaplasma phagocytophilum variants, Babesia spp., Borrelia burgdorferi, and the rickettsial endosymbiont in Ixodes scapularis (Acari: Ixodidae) from sites in Indiana, Maine, Pennsylvania, and Wisconsin.

    PubMed

    Steiner, Fresia E; Pinger, Robert R; Vann, Carolyn N; Grindle, Nate; Civitello, David; Clay, Keith; Fuqua, Clay

    2008-03-01

    In total, 394 questing adult blacklegged ticks, Ixodes scapularis Say (Acari: Ixodidae), collected at four sites were analyzed by polymerase chain reaction (PCR) for five microbial species: Anaplasma phagocytophilum, Babesia microti, Babesia odocoilei, Borrelia burgdorferi, and the rickettsial I. scapularis endosymbiont. Identities of genetic variants of A. phagocytophilum were determined by sequencing a portion of the 16S DNA. In 55% of infected ticks (193/351), a single agent was detected. In 45% (158/351), two or more agents were detected; 37% harbored two agents and 8% harbored three agents. One male tick, collected from Ft. McCoy, WI, harbored all four microbial genera The highest rates of co-infection were by the Ixodes endosymbiont and B. burgdorferi (95/351). Two species of Babesia co-occurred within a single tick population in Wells National Estuarine Research Reserve, Wells, ME, whereas only B. odocoilei was found in other tick populations. Only A. phagocytophilum human anaplasmosis variant was detected in questing ticks from Tippecanoe River State Park, IN; from Wells; and Ft. McCoy, whereas a single infected tick from Presque Isle, PA, was infected by AP-Variant 1. Partially engorged ticks from deer in Tippecanoe River State Park were all infected with AP-Variant 1. Frequency of infections with each agent varied among populations. Rates and types of co-infections were not significantly different from random except for the Ixodes endosymbiont and B. burgdorferi in male ticks, which co-occurred less frequently than expected. Thus, I. scapularis hosts an array of pathogenic and symbiotic agents and potential evidence of interactions among microbial species was observed.

  12. Serologic and molecular characterization of Anaplasma species infection in farm animals and ticks from Sicily.

    PubMed

    de la Fuente, José; Torina, Alessandra; Caracappa, Santo; Tumino, Giovanni; Furlá, Roberto; Almazán, Consuelo; Kocan, Katherine M

    2005-11-05

    Although Anaplasma marginale was known to be endemic in Italy, the diversity of Anaplasma spp. from this area have not been characterized. In this study, the prevalence of Anaplasma spp. antibodies in randomly selected farm animals collected on the island of Sicily was determined by use of a MSP5 cELISA for Anaplasma spp. and an immunofluorescence test specific for Anaplasma phagocytophilum. Genetic variation among strains of Anaplasma spp. from animals and ticks was characterized using the A. marginale msp1alpha and the Anaplasma spp. msp4 genes. Eight species of ticks were collected and tested by PCR. Seropositivity for Anaplasma spp. and A. phagocytophilum was detected in bovine and ovine samples. All the donkeys were seropositive for A. phagocytophilum but not for Anaplasma spp. Four A. marginale genotypes were identified by msp4 sequences from bovine and tick samples. Two new genotypes of Anaplasma ovis were characterized in sheep. The sequences of A. phagocytophilum from three donkeys proved to be identical to the sequence of the MRK equine isolate from California. Six A. marginale genotypes were found in cattle and one tick using the A. marginale msp1alpha sequences. All genotypes had four repeated sequences in the N-terminal portion of the MSP1a, except for one that had five repeats. The Italian strains of A. marginale contained three repeat sequences that were not reported previously. Definition of the diversity of Anaplasma spp. in Sicily reported, herein is fundamental to development of control strategies for A. marginale, A. ovis and A. phagocytophilum in Sicily.

  13. High Prevalence of Anaplasma spp. in Small Ruminants in Morocco.

    PubMed

    Ait Lbacha, H; Alali, S; Zouagui, Z; El Mamoun, L; Rhalem, A; Petit, E; Haddad, N; Gandoin, C; Boulouis, H-J; Maillard, R

    2017-02-01

    The prevalence of infection by Anaplasma spp. (including Anaplasma phagocytophilum) was determined using blood smear microscopy and PCR through screening of small ruminant blood samples collected from seven regions of Morocco. Co-infections of Anaplasma spp., Babesia spp, Theileria spp. and Mycoplasma spp. were investigated and risk factors for Anaplasma spp. infection assessed. A total of 422 small ruminant blood samples were randomly collected from 70 flocks. Individual animal (breed, age, tick burden and previous treatment) and flock data (GPS coordinate of farm, size of flock and livestock production system) were collected. Upon examination of blood smears, 375 blood samples (88.9%) were found to contain Anaplasma-like erythrocytic inclusion bodies. Upon screening with a large spectrum PCR targeting the Anaplasma 16S rRNA region, 303 (71%) samples were found to be positive. All 303 samples screened with the A. phagocytophilum-specific PCR, which targets the msp2 region, were found to be negative. Differences in prevalence were found to be statistically significant with regard to region, altitude, flock size, livestock production system, grazing system, presence of clinical cases and application of tick and tick-borne diseases prophylactic measures. Kappa analysis revealed a poor concordance between microscopy and PCR (k = 0.14). Agreement with PCR is improved by considering microscopy and packed cell volume (PCV) in parallel. The prevalence of double infections was found to be 1.7, 2.5 and 24% for Anaplasma-Babesia, Anaplasma-Mycoplasma and Anaplasma-Theileria, respectively. Co-infection with three or more haemoparasites was found in 1.6% of animals examined. In conclusion, we demonstrate the high burden of anaplasmosis in small ruminants in Morocco and the high prevalence of co-infections of tick-borne diseases. There is an urgent need to improve the control of this neglected group of diseases. © 2015 Blackwell Verlag GmbH.

  14. Molecular investigation and phylogeny of Anaplasma spp. in Mediterranean ruminants reveal the presence of neutrophil-tropic strains closely related to A. platys.

    PubMed

    Zobba, Rosanna; Anfossi, Antonio G; Pinna Parpaglia, Maria Luisa; Dore, Gian Mario; Chessa, Bernardo; Spezzigu, Antonio; Rocca, Stefano; Visco, Stefano; Pittau, Marco; Alberti, Alberto

    2014-01-01

    Few data are available on the prevalence and molecular typing of species belonging to the genus Anaplasma in Mediterranean ruminants. In this study, PCR analysis and sequencing of both 16S rRNA and groEL genes were combined to investigate the presence, prevalence, and molecular traits of Anaplasma spp. in ruminants sampled on the Island of Sardinia, chosen as a subtropical representative area. The results demonstrate a high prevalence of Anaplasma spp. in ruminants, with animals infected by at least four of six Anaplasma species (Anaplasma marginale, A. bovis, A. ovis, and A. phagocytophilum). Moreover, ruminants host a number of neutrophil-tropic strains genetically closely related to the canine pathogen A. platys. The high Anaplasma spp. prevalence and the identification of as-yet-unclassified neutrophil-tropic strains raise concerns about the specificity of serological tests routinely used in ruminants and provide additional background for reconstructing the evolutionary history of species genetically related to A. phagocytophilum.

  15. Molecular Investigation and Phylogeny of Anaplasma spp. in Mediterranean Ruminants Reveal the Presence of Neutrophil-Tropic Strains Closely Related to A. platys

    PubMed Central

    Zobba, Rosanna; Anfossi, Antonio G.; Pinna Parpaglia, Maria Luisa; Dore, Gian Mario; Chessa, Bernardo; Spezzigu, Antonio; Rocca, Stefano; Visco, Stefano; Pittau, Marco

    2014-01-01

    Few data are available on the prevalence and molecular typing of species belonging to the genus Anaplasma in Mediterranean ruminants. In this study, PCR analysis and sequencing of both 16S rRNA and groEL genes were combined to investigate the presence, prevalence, and molecular traits of Anaplasma spp. in ruminants sampled on the Island of Sardinia, chosen as a subtropical representative area. The results demonstrate a high prevalence of Anaplasma spp. in ruminants, with animals infected by at least four of six Anaplasma species (Anaplasma marginale, A. bovis, A. ovis, and A. phagocytophilum). Moreover, ruminants host a number of neutrophil-tropic strains genetically closely related to the canine pathogen A. platys. The high Anaplasma spp. prevalence and the identification of as-yet-unclassified neutrophil-tropic strains raise concerns about the specificity of serological tests routinely used in ruminants and provide additional background for reconstructing the evolutionary history of species genetically related to A. phagocytophilum. PMID:24162569

  16. Molecular survey and sequence analysis of Anaplasma spp. in cattle and ticks in a Malaysian farm.

    PubMed

    Tay, S T; Koh, F X; Kho, K L; Ong, B L

    2014-12-01

    This study was conducted to determine the occurrence of Anaplasma spp. in the blood samples of cattle, goats, deer and ticks in a Malaysian farm. Using polymerase chain reaction (PCR) and sequencing approach, Anaplasma spp. was detected from 81(84.4%) of 96 cattle blood samples. All blood samples from 23 goats and 22 deer tested were negative. Based on the analysis of the Anaplasma partial 16S ribosomal RNA gene, four sequence types (genotypes 1 to 4) were identified in this study. Genotypes 1-3 showed high sequence similarity to those of Anaplasma platys/ Anaplasma phagocytophilum, whilst genotype 4 was identical to those of Anaplasma marginale/ Anaplasma centrale/ Anaplasma ovis. Anaplasma DNA was detected from six (5.5%) of 109 ticks which were identified as Rhipicephalus (formely known as Boophilus) microplus ticks collected from the cattle. This study reported for the first time the detection of four Anaplasma sequence types circulating in the cattle population in a farm in Malaysia. The detection of Anaplasma DNA in R. microplus ticks in this study provides evidence that the ticks are one of the potential vectors for transmission of anaplasmosis in the cattle.

  17. First molecular evidence for the presence of Anaplasma DNA in milk from sheep and goats in China.

    PubMed

    Zhang, Yan; Lv, Yali; Cui, Yanyan; Wang, Jinhong; Cao, Shuxuan; Jian, Fuchun; Wang, Rongjun; Zhang, Longxian; Ning, Changshen

    2016-07-01

    Anaplasmosis, a disease caused by bacteria in the genus of Anaplasma, imposes economic constraints on animal breeders and also threatens human health. In the present study, we investigated the presence of Anaplasma spp. DNA in milk collected from sheep and goats in China. A total of 120 milk samples and 414 field-sampled blood specimens from sheep and goats were analyzed by PCR and DNA sequencing. Anaplasma ovis was detected in 12 milk samples (three from sheep and nine from goats). The blood specimens corresponding to the A. ovis DNA-positive milk were analyzed for Anaplasma DNA presence, and A. ovis DNA was identified in 10 out of the 12 blood specimens. One goat's milk sample was Anaplasma bovis DNA-positive, as was the corresponding blood sample. Anaplasma phagocytophilum was found in a milk sample and blood sample from one goat. One milk sample from Xinmi in Henan province was simultaneously positive for A. bovis and A. phagocytophilum; however, the corresponding blood was negative for both species. DNA sequencing of the PCR products and phylogenetic analysis confirmed that the sequences from the milk samples matched those of the corresponding blood samples. This is the first report to detect the Anaplasma DNA in milk samples under natural condition, and represents the first evidence of the presence of A. ovis, A. bovis and A. phagocytophilum DNA in milk from sheep and goats.

  18. Anaplasma spp. in wild mammals and Ixodes ricinus from the north of Spain.

    PubMed

    Portillo, Aránzazu; Pérez-Martínez, Laura; Santibáñez, Sonia; Santibáñez, Paula; Palomar, Ana M; Oteo, José A

    2011-01-01

    Our objectives were to investigate the presence of Anaplasma spp. infection in red deer, wild boars, and Ixodes ricinus removed from deer surveyed in La Rioja, as well as to analyze the presence of Anaplasma spp. in I. ricinus from different Spanish regions--ours included. A total of 21 deer and 13 wild boar blood samples as well as 295 I. ricinus removed from deer, vegetation, or asymptomatic people were tested by polymerase chain reaction targeting Anaplasma spp. 16S rRNA gene and groESL heat shock operon. Twelve deer blood samples were found to be infected with Anaplasma centrale (n = 7) or Anaplasma phagocytophilum (n = 5). No wild boar blood samples gave positive polymerase chain reaction results. Further, A. phagocytophilum was detected in 12 out of 89 I. ricinus removed from deer and in 18 out of 168 I. ricinus collected over vegetation in the North of Spain. Anaplasma spp. was not detected in any of the 38 I. ricinus removed from people. Nucleotide sequences for 16S rRNA gene showed substancial heterogeneity. The etiological agent of human anaplasmosis was found in two deer blood samples, an adult tick from deer, and a nymph from vegetation. The 16S rRNA sequences for 12 out of 35 samples matched the sequence of the Ap-variant 1 strain previously described in the United States, and the remaining 19 positive samples (deer blood and I. ricinus) showed variations with unknown significance. Although the groEL DNA sequences varied among analyzed strains, the deduced amino acid sequences did not change for any of them. This study suggests that deer population from La Rioja harbors strains of A. phagocytophilum similar to that pathogen for humans and other of unknown pathogenicity. Further, it seems that the Ap-variant 1 is circulating among I. ricinus ticks from the North of Spain more frequently than the A. phagocytophilum strain associated to human anaplasmosis.

  19. Molecular detection of Anaplasma spp. in pangolins (Manis javanica) and wild boars (Sus scrofa) in Peninsular Malaysia.

    PubMed

    Koh, Fui Xian; Kho, Kai Ling; Panchadcharam, Chandrawathani; Sitam, Frankie Thomas; Tay, Sun Tee

    2016-08-30

    Anaplasma spp. infects a wide variety of wildlife and domestic animals. This study describes the identification of a novel species of Anaplasma (Candidatus Anaplasma pangolinii) from pangolins (Manis javanica) and Anaplasma bovis from wild boars (Sus scrofa) in Malaysia. Based on 16S rRNA gene sequences, Candidatus Anaplasma pangolinii is identified in a distinct branch within the family Anaplasmataceae, exhibiting the closest sequence similarity with the type strains of Anaplasma bovis (97.7%) and Anaplasma phagocytophilum (97.6%). The sequence also aligned closely (99.9%) with that of an Anaplasma spp. (strain AnAj360) detected from Amblyomma javanense ticks. The nearly full length sequence of the 16S rRNA gene derived from two wild boars in this study demonstrated the highest sequence similarity (99.7%) to the A. bovis type strain. Partial 16S rRNA gene fragments of A. bovis were also detected from a small population of Haemaphysalis bispinosa cattle ticks in this study. Our finding suggests a possible spread of two Anaplasma species in the Malaysian wildlife and ticks. The zoonotic potential of the Anaplasma species identified in this study is yet to be determined.

  20. Prevalence and genotypes of Anaplasma species and habitat suitability for ticks in a Mediterranean ecosystem.

    PubMed

    Torina, Alessandra; Alongi, Angelina; Naranjo, Victoria; Estrada-Peña, Agustín; Vicente, Joaquín; Scimeca, Salvatore; Marino, Anna M F; Salina, Felice; Caracappa, Santo; de la Fuente, José

    2008-12-01

    Anaplasma species are tick-transmitted pathogens that impact veterinary and human health. Sicily is one of the locations where these pathogens are endemic. Sicily represents a typical Mediterranean ecosystem to study Anaplasma infection and tick habitat suitability. The aims of this study were (i) to characterize by 16S rRNA and species-specific msp4 gene PCR the prevalence and genotypes of A. marginale, A. phagocytophilum, and A. ovis in the most abundant host species in Sicilian provinces and (ii) to correlate differences between hosts and between western and eastern Sicily with the habitat suitability for ticks in these regions. Differences were found in the prevalence of Anaplasma spp. between different hosts and between western and eastern provinces. The differences in Anaplasma prevalence between different hosts may be explained by pathogen host tropism. The differences between western and eastern provinces correlated with the tick habitat suitability in these regions. The analysis of Anaplasma genotypes suggested a higher host and regional specificity for A. phagocytophilum than for A. marginale and A. ovis strains, a finding probably associated with the broader host range of A. phagocytophilum. The presence of identical A. marginale genotypes in the two regions may reflect cattle movement. The results for A. ovis suggested the possibility of some genotypes being host specific. These results provide information potentially useful for the management of tick-borne diseases caused by Anaplasma spp. in Sicily and other Mediterranean regions and may contribute to the development of models to predict the risks for these tick-borne pathogens.

  1. Molecular and phylogenetic analysis of Anaplasma spp. in sheep and goats from six provinces of China.

    PubMed

    Zhang, Yan; Lv, Yali; Zhang, Feifei; Zhang, Wenjing; Wang, Jinhong; Cui, Yanyan; Wang, Rongjun; Jian, Fuchun; Zhang, Longxian; Ning, Changshen

    2016-12-30

    Members of the genus Anaplasma are important emerging tick-borne pathogens in both humans and animals in tropical and subtropical areas. Here, we investigated the presence of Anaplasma spp. in 621 sheep and 710 goats from six provinces of China. Polymerase chain reaction (PCR) and DNA sequencing were conducted to determine the prevalence of Anaplasma (A.) phagocytophilum, A. ovis and A. bovis targeting the 16S ribosomal RNA or the major surface protein 4 gene. PCR revealed Anaplasma in 39.0% (240/621) of sheep and 45.5% (323/710) of goats. The most frequently detected species was A. ovis (88/621, 14.2% for sheep; 129/710, 18.2% for goats), followed by A. bovis (60/621, 9.7% for sheep; 74/710, 10.4% for goats) and A. phagocytophilum (33/621, 5.3% for sheep; 15/710, 2.1% for goats). Additionally, eight sheep and 20 goats were found to be infected with three pathogens simultaneously. DNA sequencing confirmed the presence of these three Anaplasma species in the investigated areas, and phylogenetic analysis indicated that there was geographic segregation to a certain extent, as well as a relationship between the host and cluster of A. ovis. The results of the present study provide valuable data that helps understand the epidemiology of anaplasmosis in ruminants from China.

  2. Molecular and phylogenetic analysis of Anaplasma spp. in sheep and goats from six provinces of China

    PubMed Central

    Zhang, Yan; Lv, Yali; Zhang, Feifei; Zhang, Wenjing; Wang, Jinhong; Cui, Yanyan; Wang, Rongjun; Jian, Fuchun; Zhang, Longxian

    2016-01-01

    Members of the genus Anaplasma are important emerging tick-borne pathogens in both humans and animals in tropical and subtropical areas. Here, we investigated the presence of Anaplasma spp. in 621 sheep and 710 goats from six provinces of China. Polymerase chain reaction (PCR) and DNA sequencing were conducted to determine the prevalence of Anaplasma (A.) phagocytophilum, A. ovis and A. bovis targeting the 16S ribosomal RNA or the major surface protein 4 gene. PCR revealed Anaplasma in 39.0% (240/621) of sheep and 45.5% (323/710) of goats. The most frequently detected species was A. ovis (88/621, 14.2% for sheep; 129/710, 18.2% for goats), followed by A. bovis (60/621, 9.7% for sheep; 74/710, 10.4% for goats) and A. phagocytophilum (33/621, 5.3% for sheep; 15/710, 2.1% for goats). Additionally, eight sheep and 20 goats were found to be infected with three pathogens simultaneously. DNA sequencing confirmed the presence of these three Anaplasma species in the investigated areas, and phylogenetic analysis indicated that there was geographic segregation to a certain extent, as well as a relationship between the host and cluster of A. ovis. The results of the present study provide valuable data that helps understand the epidemiology of anaplasmosis in ruminants from China. PMID:27456776

  3. Anaplasma phagocytophilum—a widespread multi-host pathogen with highly adaptive strategies

    PubMed Central

    Stuen, Snorre; Granquist, Erik G.; Silaghi, Cornelia

    2013-01-01

    The bacterium Anaplasma phagocytophilum has for decades been known to cause the disease tick-borne fever (TBF) in domestic ruminants in Ixodes ricinus-infested areas in northern Europe. In recent years, the bacterium has been found associated with Ixodes-tick species more or less worldwide on the northern hemisphere. A. phagocytophilum has a broad host range and may cause severe disease in several mammalian species, including humans. However, the clinical symptoms vary from subclinical to fatal conditions, and considerable underreporting of clinical incidents is suspected in both human and veterinary medicine. Several variants of A. phagocytophilum have been genetically characterized. Identification and stratification into phylogenetic subfamilies has been based on cell culturing, experimental infections, PCR, and sequencing techniques. However, few genome sequences have been completed so far, thus observations on biological, ecological, and pathological differences between genotypes of the bacterium, have yet to be elucidated by molecular and experimental infection studies. The natural transmission cycles of various A. phagocytophilum variants, the involvement of their respective hosts and vectors involved, in particular the zoonotic potential, have to be unraveled. A. phagocytophilum is able to persist between seasons of tick activity in several mammalian species and movement of hosts and infected ticks on migrating animals or birds may spread the bacterium. In the present review, we focus on the ecology and epidemiology of A. phagocytophilum, especially the role of wildlife in contribution to the spread and sustainability of the infection in domestic livestock and humans. PMID:23885337

  4. The occurrence of Anaplasma phagocyto- philum in wild bison from the Bialowieza Primeval Forest in Eastern Poland.

    PubMed

    Dzięgiel, Beata; Adaszekl, Łukasz; Krzysiak, Michał; Skrzypczak, Maciej; Adaszek, Michał; Furmaga, Barbara; Winiarczyk, Stanislaw

    2015-01-01

    The aim of the present study was to investigate the occurrence of Anaplasma spp. in a group of 120 wild bison (Bison bonasus) from the Bialowieza Primeval Forest in eastern Poland and to determine which species of Anaplasma could infect these animals based on a PCR of a part of the 16S rRNA gene followed by sequencing. The PCR technique showed the presence of 16S rRNA Anaplasma spp. genetic material in the blood of 22 from a total of 120 animals. DNA amplification by means of the primers EHR 521 and EHR 747 gave a product size of 252-bp. The sequences of the PCR products obtained showed 100% homology with each other and 100% homology with the Anaplasma phagocytophilum GU 183908 sequence from our earlier study, isolated from a horse with a clinical case of anaplasmosis. The similarity of the sequences with the sequences of the 16S rRNA gene isolated from various Anaplasma species deposited in the GeneBank, ranged between 95.8% and 98.8%. Based on the results of molecular analysis, bacterial DNA detected in the blood of 22 wild bison was identified as Anaplasma phagocytophilum.

  5. Migratory and carnivorous birds in Brazil: reservoirs for Anaplasma and Ehrlichia species?

    PubMed

    Machado, Rosangela Zacarias; André, Marcos Rogério; Werther, Karin; de Sousa, Eliane; Gavioli, Fernando Antônio; Alves Junior, José Roberto Ferreira

    2012-08-01

    In order to investigate new hosts for Anaplasmataceae agents in Brazil, we collected blood samples from 21 wild birds. Using molecular techniques, we detected the presence of Anaplasma phagocytophilum, Ehrlichia chaffeensis, and an Ehrlichia species closely related to Ehrlichia canis in carnivorous avian blood samples. In addition, an Ehrlichia species closely related to an Ehrlichia species found in wild felines in Brazil was also detected in a goose blood sample. Wild birds may play a role as carriers of Anaplasmataceae agents in Brazil.

  6. Molecular Detection of Anaplasma spp. and Ehrlichia spp. in Ruminants from Twelve Provinces of China

    PubMed Central

    Qiu, Haixiang; Kelly, Patrick John; Zhang, Jilei; Luo, Qinghua; Yang, Yi; Mao, Yongjiang; Yang, Zhangping; Li, Jing; Wu, Hongzhuan

    2016-01-01

    Anaplasma spp. and Ehrlichia spp. are tick-transmitted bacteria that are of significant economic importance as they can infect large and small ruminants and also people. There is little information on anaplasmosis and ehrlichiosis in ruminants in China. 16S rRNA FRET-qPCRs were used to screen convenience whole blood samples from 2,240 domestic ruminants in 12 provinces of China for Anaplasma spp. and Ehrlichia spp. Positive samples were further analyzed with a standard PCR for the gltA. Anaplasma spp. DNA was detected in the sheep (11.7%; 13/111), goats (81.8%; 219/270), cattle (13.2%; 241/1,830), and water buffaloes (6.9%; 2/29). Ehrlichia spp. DNA was detected in sheep (1.8%; 2/111), goats (1.1%; 3/270), and cattle (3.6%; 65/1830) but not in water buffaloes (0/29). Sequencing of gltA PCR products showed that A. marginale, A. ovis, Ehrlichia canis, and Ehrlichia sp. (JX629807) were present in ruminants from China, while the 16S rRNA FRET-qPCR sequence data indicated that there might also be A. platys, A. phagocytophilum, Anaplasma sp. BL126-13 (KJ410243), and Anaplasma sp. JC3-6 (KM227012). Our study shows that domestic ruminants from China are not uncommonly infected with a variety of Anaplasma spp. and Ehrlichia spp. PMID:28096822

  7. Anaplasma platys Immunoblot Test Using Major Surface Antigens.

    PubMed

    Lai, Tzung-Huei; Parraga, Maria E; Alvarez, Elizabeth; Rikihisa, Yasuko

    2016-09-01

    Anaplasma platys is an uncultivable tick-borne obligatory intracellular bacterium, which is known to infect platelets of dogs. A. platys causes infectious canine cyclic thrombocytopenia in subtropical and tropical regions throughout the world. Several cases of human infection with A. platys infection have also been reported. However, seroprevalence of A. platys exposure and infection has not been determined in most of the regions, in part, due to lack of a simple and reliable assay method. Furthermore, A. platys antigens recognized by dogs are unknown. We previously sequenced gene encoding A. platys major outer membrane proteins P44 and Omp-1X. In the present study, we obtained purified recombinant A. platys P44 and Omp-1X proteins, and using them as antigens in immunoblotting examined seroreactivity in dogs. Of 34 specimens from Venezuela where A. platys infection was previously reported, 25 specimens (73.5%) reacted to rAplP44 and/or rAplOMP-1X. Neither Anaplasma phagocytophilum-seropositive (N = 10) nor A. phagocytophilum-seronegative canine specimens (N = 10) from the geographic regions where A. platys infection has never been reported, reacted rAplP44 or rAplOMP-1X. The result indicates a high A. platys seroprevalence rate in tested dogs from Venezuela and suggests that the immunoblot analysis based on recombinant A. platys major outer membrane proteins can provide a simple and defined tool to enlighten the prevalence of A. platys infection.

  8. Assessment of a quantitative 5' nuclease real-time polymerase chain reaction using groEL gene for Ehrlichia and Anaplasma species in rodents in Brazil.

    PubMed

    Benevenute, Jyan Lucas; Dumler, John Stephen; Ogrzewalska, Maria; Roque, André Luiz Rodrigues; Mello, Victoria Valente Califre; de Sousa, Keyla Carstens Marques; Gonçalves, Luiz Ricardo; D'Andrea, Paulo Sérgio; de Sampaio Lemos, Elba Regina; Machado, Rosangela Zacarias; André, Marcos Rogério

    2017-06-01

    New genotypes of Anaplasmataceae agents have been detected in wild carnivores, birds and deer in Brazil. The present work aimed to investigate the presence of Ehrlichia and Anaplasma species in rodents sampled in Brazil. Additionally, a newly designed quantitative 5' nuclease real-time multiplex PCR for Ehrlichia and Anaplasma spp. detection based on groEL gene amplification was designed, showing high specificity and sensitivity (10 groEL fragment copy/μL). Between 2000 and 2011, different rodent species [n=60] were trapped in 5 Brazilian biomes. Among 458 rodent spleen samples, 0.4% (2/458) and 2.4% (11/458) were positive for Ehrlichia and Anaplasma spp., respectively. Of 458 samples, 2.0% (9/458) and 1.1% (5/458) were positive for Anaplasma sp. and Ehrlichia sp., respectively, using conventional 16S rRNA PCR assays. Maximum Likelihood phylogenetic analyse based on a small region of 16S rRNA genes positioned the Anaplasma genotypes in rodents near Anaplasma phagocytophilum or Anaplasma marginale and Anaplasma odocoilei isolates. Ehrlichia genotypes were closely related to E. canis. There was a low occurrence of Anaplasma and Ehrlichia in wild and synanthropic rodents in Brazil, suggesting the circulation of new genotypes of these agents in rodents in the studied areas. Copyright © 2017 Elsevier GmbH. All rights reserved.

  9. Factors associated with Anaplasma spp. seroprevalence among dogs in the United States.

    PubMed

    McMahan, Christopher S; Wang, Dongmei; Beall, Melissa J; Bowman, Dwight D; Little, Susan E; Pithua, Patrick O; Sharp, Julia L; Stich, Roger W; Yabsley, Michael J; Lund, Robert B

    2016-03-22

    Dogs in the United States are hosts to a diverse range of ticks and tick-borne pathogens, including A. phagocytophilum, an important emerging canine and human pathogen. Previously, a Companion Animal Parasite Council (CAPC)-sponsored workshop proposed factors purported to be associated with the infection risk for tick-transmitted pathogens in dogs in the United States, including climate conditions, socioeconomic characteristics, local topography, and vector distribution. Approximately four million test results from routine veterinary diagnostic tests from 2011-2013, which were collected on a county level across the contiguous United States, are statistically analyzed with the proposed factors via logistic regression and generalized estimating equations. Spatial prevalence maps of baseline Anaplasma spp. prevalence are constructed from Kriging and head-banging smoothing methods. All of the examined factors, with the exception of surface water coverage, were significantly associated with Anaplasma spp. prevalence. Overall, Anaplasma spp. prevalence increases with increasing precipitation and forestation coverage and decreases with increasing temperature, population density, relative humidity, and elevation. Interestingly, socioeconomic status and deer/vehicle collisions were positively and negatively correlated with canine Anaplasma seroprevalence, respectively. A spatial map of the canine Anaplasma hazard is an auxiliary product of the analysis. Anaplasma spp. prevalence is highest in New England and the Upper Midwest. The results from the two posited statistical models (one that contains an endemic areas assumption and one that does not) are in general agreement, with the major difference being that the endemic areas model estimates a larger prevalence in Western Texas, New Mexico, and Colorado. As A. phagocytophilum is zoonotic, the results of this analysis could also help predict areas of high risk for human exposure to this pathogen.

  10. Anaplasma infections in ticks and reservoir host from Slovakia.

    PubMed

    Víchová, Bronislava; Majláthová, Viktória; Nováková, Mária; Stanko, Michal; Hviščová, Ivana; Pangrácová, Lucia; Chrudimský, Tomáš; Čurlík, Ján; Petko, Branislav

    2014-03-01

    Anaplasma phagocytophilum is a worldwide distributed bacterium with a significant medical and veterinary importance. It grows within the phagosome of infected neutrophils and is responsible for human granulocytic anaplasmosis (HGA), tick-borne fever (TBF) of small ruminants and cattle, canine and equine granulocytic anaplasmosis, but infects also a great variety of wildlife species. Wild ungulates and rodents are considered reservoirs of infection in natural foci. The objective of this study was to determine the spectrum of animal species involved in the circulation of A. phagocytophilum in Slovakia and to analyze the variability of obtained nucleotide sequences, in order to determine whether genotypes from Slovakia cluster according to host-species or geographical location. Several animal species and vector ticks were screened for the presence of members of the family Anaplasmataceae using PCR based methods. Additional data on the molecular evidence of Anaplasma ovis and Candidatus Neoehrlichia mikurensis are presented. These pathogens were detected in tested sheep flocks and rodents with the mean infection rates of 8.16% and 10.75%, respectively. A. phagocytophilum was genotyped by 16S rRNA and groEL gene sequencing. Bacterial DNA was confirmed in questing ixodid ticks, in domesticated canine, wild rodents and several species of wild ungulates. In European isolates, 16S rRNA gene does not seem to be an appropriate locus for the analyses of heterogeneity as it is too conservative. Similarly, 16S rRNA isolates from our study did not reveal any polymorphisms. All isolates were identical in overlapped region and showed identity with sequences from ticks, horses or ruminants previously isolated elsewhere in the world. On the other hand, the groESL heat shock operon is widely used for determination of diversity and the analyses have already revealed considerable degree of heterogeneity. Tested ungulates were infected with A. phagocytophilum to a considerable extent

  11. The prevalence of Anaplasma platys and a potential novel Anaplasma species exceed that of Ehrlichia canis in asymptomatic dogs and Rhipicephalus sanguineus in Taiwan.

    PubMed

    Yuasa, Yumi; Tsai, Yi-Lun; Chang, Chao-Chin; Hsu, Tien-Huan; Chou, Chi-Chung

    2017-09-12

    Canine anaplasmosis is regarded as an infection by Anaplasma platys rather than zoonotic Anaplasma phagocytophilum in subtropical areas based on the assumption that the common dog tick species is Rhipicephalus sanguineus, which transmits E. canis and presumably A. platys. We investigated asymptomatic dogs and dog ticks from 16 communities in Nantou County, Taiwan to identify common dog tick species and to determine the prevalence of Anaplasma and Ehrlichia spp. Of total 175 canine blood samples and 315 ticks, including 306 R. sanguineus and 9 Haemaphysalis hystricis, 15 dogs and 3 R. sanguineus ticks were positive for E. canis, while 47 dogs and 71 R. sanguineus ticks were positive for A. platys, via nested PCR for 16S rDNA and DNA sequencing of selected positive amplicons. However, among the dogs and ticks that were positive to A. platys 16S rDNA, only 20 dogs and 11 ticks were positive to nested PCR for A. platys groEL gene. These results revealed the importance of searching for novel Anaplasma spp. closely related to A. platys in dogs and ticks. Seropositivity to a commercial immunochromatographic test SNAP 4Dx Anaplasma sp. was not significantly associated with PCR positivity for A. platys but with infestation by ticks carrying A. platys (P<0.05). Accordingly, R. sanguineus may be involved in transmission of A. platys but may not act as a reservoir of E. canis and PCR results for 16S rDNA could be a problematic diagnostic index for A. platys infection.

  12. A Molecular and Serological Survey of Rickettsiales Bacteria in Wild Sika Deer (Cervus nippon nippon) in Shizuoka Prefecture, Japan: High Prevalence of Anaplasma Species.

    PubMed

    Wu, Dongxing; Wuritu; Yoshikawa, Yuko; Gaowa; Kawamori, Fumihiko; Ikegaya, Asaka; Ohtake, Masayoshi; Ohashi, Masataka; Shimada, Masahiko; Takada, Ayumi; Iwai, Katsuki; Ohashi, Norio

    2015-01-01

    We surveyed Rickettsiales bacteria, including Rickettsia, Ehrlichia, Anaplasma, and Neoehrlichia, in wild sika deer (Cervus nippon nippon) from Shizuoka prefecture, Japan. In spleen samples from 187 deer, Anaplasma phagocytophilum (deer type), A. bovis, and A. centrale were successfully detected by PCR assay targeting to 16S rDNA or p44/msp2, and their positive rates were 96.3% (180/187), 53.5% (100/187), and 78.1% (146/187), respectively. Additionally, 2 or 3 Anaplasma species could be detected from a single deer in 165 spleen samples (88.2%), indicating dual or triple infection. In contrast, A. phagocytophilum (human type) 16S rDNA, Rickettsia gltA, Ehrlichia p28/omp-1, and Neoehrlichia 16S rDNA could not be amplified. The serological test of 105 deer serum samples by immunofluorescence assay showed that the detection of antibodies against antigens of A. phagocytophilum HZ (US-human isolate) and Rickettsia japonica YH were 29.5% (31/105) and 75.2% (79/105), respectively. These findings suggest that A. phagocytophilum (deer type), A. centrale, and A. bovis are highly dominant and prevalent in wild sika deer from Shizuoka, a central region of Japan, and that the antibodies against some Rickettsiales bacteria have also been retained in deer blood.

  13. A Molecular survey of Anaplasma spp., Rickettsia spp., Ehrlichia canis and Babesia microti in foxes and fleas from Sicily.

    PubMed

    Torina, A; Blanda, V; Antoci, F; Scimeca, S; D'Agostino, R; Scariano, E; Piazza, A; Galluzzo, P; Giudice, E; Caracappa, S

    2013-11-01

    Fleas (Insecta: Siphonaptera) are obligate bloodsucking insects, which parasitize birds and mammals, and are distributed throughout the world. Several species have been implicated in pathogen transmission. This study aimed to monitor red foxes and the fleas isolated from them in the Palermo and Ragusa provinces of Sicily, Italy, as these organisms are potential reservoirs and vectors of pathogens. Thirteen foxes (Vulpes vulpes) and 110 fleas were analysed by polymerase chain reaction (PCR) to detect DNA of the pathogens Ehrlichia canis, Babesia microti, Rickettsia spp., Anaplasma phagocytophilum, Anaplasma platys, Anaplasma marginale and Anaplasma ovis. In the foxes, A. ovis was detected in only one animal, whereas the prevalence of the E. canis pathogen was 31%. B. microti and Rickettsia spp. were not detected. Of all of the collected fleas, 75 belonged to the species Xenopsylla cheopis, 32 belonged to Ctenocephalides canis, two belonged to Ctenocephalides felis and one belonged to Cediopsylla inaequalis. In the fleas, the following pathogens were found: A. ovis (prevalence 25%), A. marginale (1%), A. phagocytophilum (1%), Rickettsia felis (2%) and E. canis (3%). X. cheopis was the flea species most frequently infected with Anaplasma, in particular A. ovis (33%), A. marginale (1%) and A. phagocytophilum (1%). Both C. felis exemplars were positive for R. felis. E. canis was found in the lone C. inaequalis and also in 3% of the X. cheopis specimens. No fleas were positive for B. microti or A. platys. As foxes often live in proximity to domestic areas, they may constitute potential reservoirs for human and animal parasites. Further studies should be performed on fleas to determine their vectorial capacity. © 2013 Blackwell Verlag GmbH.

  14. Genetic characterization of Anaplasma ovis strains from bighorn sheep in Montana.

    PubMed

    de la Fuente, José; Atkinson, Mark W; Hogg, John T; Miller, David S; Naranjo, Victoria; Almazán, Consuelo; Anderson, Neil; Kocan, Katherine M

    2006-04-01

    Wildlife reservoir species and genetic diversity of Anaplasma ovis (Rickettsiales: Anaplasmataceae) have been poorly characterized. Bighorn sheep (Ovis canadensis), captured in Montana from December 2004 to January 2005, were tested for antibodies to Anaplasma spp.; the presence of A. ovis was determined by the characterization of major surface protein msp4 sequences. Anaplasma antibodies were detected in 25/180 (14%) sampled bighorn sheep and A. ovis msp4 sequences were amplified by polymerase chain reaction (PCR) and sequenced from 9/23 (39%) of seropositive animals. All animals were negative by PCR for the related pathogens, Anaplasma phagocytophilum and Anaplasma marginale. All msp4 sequences identified in the bighorn sheep were identical and corresponded to a single A. ovis genotype that was identical to a sheep isolate reported previously from Idaho. The finding of a single genotype of A. ovis in this wild herd of bighorn sheep was in contrast to the genetic diversity reported for A. marginale in cattle herds in the western United States and worldwide. These results demonstrated that bighorn sheep may be a wildlife reservoir of A. ovis in Montana.

  15. Prevalence of Anaplasma and Bartonella spp. in Ticks Collected from Korean Water Deer (Hydropotes inermis argyropus)

    PubMed Central

    Kang, Jun-Gu; Ko, Sungjin; Kim, Heung-Chul; Chong, Sung-Tae; Klein, Terry A.; Chae, Jeong-Byoung; Jo, Yong-Sun; Choi, Kyoung-Seong; Yu, Do-Hyeon; Park, Bae-Keun; Park, Jinho; Chae, Joon-Seok

    2016-01-01

    Deer serve as reservoirs of tick-borne pathogens that impact on medical and veterinary health worldwide. In the Republic of Korea, the population of Korean water deer (KWD, Hydropotes inermis argyropus) has greatly increased from 1982 to 2011, in part, as a result of reforestation programs established following the Korean War when much of the land was barren of trees. Eighty seven Haemaphysalis flava, 228 Haemaphysalis longicornis, 8 Ixodes nipponensis, and 40 Ixodes persulcatus (21 larvae, 114 nymphs, and 228 adults) were collected from 27 out of 70 KWD. A total of 89/363 ticks (266 pools, 24.5% minimum infection rate) and 5 (1.4%) fed ticks were positive for Anaplasma phagocytophilum using nested PCR targeting the 16S rRNA and groEL genes, respectively. The 16S rRNA gene fragment sequences of 88/89 (98.9%) of positive samples for A. phagocytophilum corresponded to previously described gene sequences from KWD spleen tissues. The 16S rRNA gene fragment sequences of 20/363 (5.5%) of the ticks were positive for A. bovis and were identical to previously reported sequences. Using the ITS specific nested PCR, 11/363 (3.0%) of the ticks were positive for Bartonella spp. This is the first report of Anaplasma and Bartonella spp. detected in ticks collected from KWD, suggesting that ticks are vectors of Anaplasma and Bartonella spp. between reservoir hosts in natural surroundings. PMID:26951985

  16. Prevalence of Anaplasma and Bartonella spp. in Ticks Collected from Korean Water Deer (Hydropotes inermis argyropus).

    PubMed

    Kang, Jun-Gu; Ko, Sungjin; Kim, Heung-Chul; Chong, Sung-Tae; Klein, Terry A; Chae, Jeong-Byoung; Jo, Yong-Sun; Choi, Kyoung-Seong; Yu, Do-Hyeon; Park, Bae-Keun; Park, Jinho; Chae, Joon-Seok

    2016-02-01

    Deer serve as reservoirs of tick-borne pathogens that impact on medical and veterinary health worldwide. In the Republic of Korea, the population of Korean water deer (KWD, Hydropotes inermis argyropus) has greatly increased from 1982 to 2011, in part, as a result of reforestation programs established following the Korean War when much of the land was barren of trees. Eighty seven Haemaphysalis flava, 228 Haemaphysalis longicornis, 8 Ixodes nipponensis, and 40 Ixodes persulcatus (21 larvae, 114 nymphs, and 228 adults) were collected from 27 out of 70 KWD. A total of 89/363 ticks (266 pools, 24.5% minimum infection rate) and 5 (1.4%) fed ticks were positive for Anaplasma phagocytophilum using nested PCR targeting the 16S rRNA and groEL genes, respectively. The 16S rRNA gene fragment sequences of 88/89 (98.9%) of positive samples for A. phagocytophilum corresponded to previously described gene sequences from KWD spleen tissues. The 16S rRNA gene fragment sequences of 20/363 (5.5%) of the ticks were positive for A. bovis and were identical to previously reported sequences. Using the ITS specific nested PCR, 11/363 (3.0%) of the ticks were positive for Bartonella spp. This is the first report of Anaplasma and Bartonella spp. detected in ticks collected from KWD, suggesting that ticks are vectors of Anaplasma and Bartonella spp. between reservoir hosts in natural surroundings.

  17. Molecular detection of novel Anaplasmataceae closely related to Anaplasma platys and Ehrlichia canis in the dromedary camel (Camelus dromedarius).

    PubMed

    Bastos, Armanda D S; Mohammed, Osama B; Bennett, Nigel C; Petevinos, Charalambos; Alagaili, Abdulaziz N

    2015-09-30

    Serological surveys have confirmed Anaplasma marginale and Anaplasma phagocytophilum infections in dromedary camels, but molecular surveys and genetic characterisation of camel-associated Anaplasma species are lacking. In this study, we detected tick-borne Anaplasmataceae in 30 of 100 (30%) healthy dromedary camels screened using a combined 16S rRNA-groEL PCR-sequencing approach. Nucleotide sequencing confirmed Anaplasmataceae genome presence in 28 of the 33 16S rRNA PCR-positive samples, with two additional positive samples, for which 16S rRNA sequence data were ambiguous, being identified by groEL gene characterisation. Phylogenetic analyses of a 1289 nt segment of the 16S rRNA gene confirmed the presence of a unique Ehrlichia lineage and a discrete Anaplasma lineage, comprising three variants, occurring at an overall prevalence of 4% and 26%, respectively. Genetic characterisation of an aligned 559 nt groEL gene region revealed the camel-associated Anaplasma and Ehrlichia lineages to be novel and most closely related to Anaplasma platys and Ehrlichia canis. Based on the confirmed monophyly, minimum pairwise genetic distances between each novel lineage and its closest sister taxon, and the inability to isolate the bacteria, we propose that Candidatus status be assigned to each. This first genetic characterisation of Anaplasmataceae from naturally infected, asymptomatic dromedary camels in Saudi Arabia confirms the presence of two novel lineages that are phylogenetically linked to two pathogenic canid species of increasing zoonotic concern.

  18. Detection of A. phagocytophilum and E. chaffeensis in patient and mouse blood and ticks by a duplex real-time PCR assay.

    PubMed

    Dong, Tuo; Qu, Zhangyi; Zhang, Lijuan

    2013-01-01

    Human granulocytic anaplasmosis (HGA) and human monocytic ehrlichiosis (HME) are emerging, tick-borne, zoonotic infectious diseases caused by Anaplasma phagocytophilum and Ehrlichia chaffeensis, respectively. Early diagnosis is essential for rapid clinical treatment to avoid misdiagnosis and severe patient outcomes. Simple, sensitive and reliable diagnostic methods are urgently needed. In this study, we developed a duplex real-time PCR assay targeting the A. phagocytophilum ankA gene and the E. chaffeensis TRP120 gene, respectively. The lowest limit of detection of the duplex real-time PCR assay was 100 copies of the targeted A. phagocytophilum ankA gene and the E. chaffeensis TRP120 gene per reaction, and the specificity was 100%. Detection in blood DNA samples from the acute stage of illness for 22 HGA cases and 8 HME cases indicated that the duplex real-time PCR assay was more sensitive than the nested PCR assay. The infection of Citellusundulatus Pallas with A. phagocytophilum and E. chaffeensis was first confirmed in Xinjiang Province and the positive rate was 3.1% for A. phagocytophilum, 6.3% for E. chaffeensis and 3.1% for co-infection with both pathogens. The rates of A. phagocytophilum and E. chaffeensis infection of D. silvarum ticks collected from Shanxi Province were 8.2% and 14.8%, respectively, and the co-infection rate was 3.3%. The rates of A. phagocytophilum and E. chaffeensis infection in H. longicornis ticks collected from Shandong Province were 1.6% and 6.3%, respectively, and the co-infection rate was 1.6%.

  19. Evaluation of Gulf Coast Ticks (Acari: Ixodidae) for Ehrlichia and Anaplasma Species.

    PubMed

    Allerdice, Michelle E J; Hecht, Joy A; Karpathy, Sandor E; Paddock, Christopher D

    2017-03-01

    Amblyomma maculatum Koch (the Gulf Coast tick) is an aggressive, human-biting ixodid tick distributed throughout much of the southeastern United States and is the primary vector for Rickettsia parkeri, an emerging human pathogen. Amblyomma maculatum has diverse host preferences that include white-tailed deer, a known reservoir for Ehrlichia and Anaplasma species, including the human pathogens E. ewingii and E. chaffeensis. To examine more closely the potential role of A. maculatum in the maintenance of various pathogenic Ehrlichia and Anaplasma species, we screened DNA samples from 493 questing adult A. maculatum collected from six U.S. states using broad-range Anaplasmataceae and Ehrlichia genus-specific PCR assays. Of the samples tested, four (0.8%) were positive for DNA of Ehrlichia ewingii, one (0.2%) was positive for Anaplasma platys, and one (0.2%) was positive for a previously unreported Ehrlichia species closely related to Ehrlichia muris and an uncultivated Ehrlichia species from Haemaphysalis longicornis ticks in Japan. No ticks contained DNA of Ehrlichia chaffeensis, Ehrlichia canis, the Panola Mountain Ehrlichia, or Anaplasma phagocytophilum. This is the first identification of E. ewingii, A. platys, and the novel Ehrlichia in questing Gulf Coast ticks; nonetheless the low prevalence of these agents suggests that A. maculatum is not likely an important vector of these zoonotic pathogens.

  20. Molecular survey of Anaplasma and Ehrlichia infections of feral raccoons (Procyon lotor) in Hokkaido, Japan.

    PubMed

    Sashika, Mariko; Abe, Go; Matsumoto, Kotaro; Inokuma, Hisashi

    2011-04-01

    Infection by Anaplasma and Ehrlichia in feral raccoons (Procyon lotor) in Hokkaido, Japan, was examined by molecular methods. A polymerase chain reaction (PCR) screen for Anaplasmataceae, based on 16S rRNA, showed that 38 (5.4%) of 699 raccoons examined were positive. These 38 positive samples were examined for Anaplasma phagocytophilum, Anaplasma bovis, Ehrlichia chaffeensis, and Ehrlichia canis infection by species-specific nested PCR. Nested PCR results indicated that 36 of the 38 samples were positive for A. bovis. All 38 samples were PCR negative for A. phagocytophilum, E. chaffeensis, and E. canis. This is the first report of the detection of A. bovis in the peripheral blood of raccoons. A total of 124 raccoons were infested with ticks, including Ixodes ovatus, Ixodes persulcatus, and Haemaphysalis spp. The rate of A. bovis infection in raccoons infested with Haemaphysalis spp. (46.7%, 7/15) was significantly higher than that in raccoons without Haemaphysalis spp. infestation (3.7%, 4/109, p < 0.001). No significant differences were observed in A. bovis infection rates between raccoons infested with I. ovatus or I. persulcatus and those not so infested. A total of four ticks (two males and two nymphs) and one larval pools from four raccoons showed positive for A. bovis-specific nested PCR. This results support the correlation between the A. bovis infection of raccoons and Haemaphysalis infestation. In conclusion, raccoons could be possible reservoir animals for A. bovis, and A. bovis infection in raccoons may be related to infestation with Haemaphysalis spp.

  1. Sequence analysis of the msp4 gene of Anaplasma ovis strains.

    PubMed

    de la Fuente, José; Atkinson, Mark W; Naranjo, Victoria; Fernández de Mera, Isabel G; Mangold, Atilio J; Keating, Kimberly A; Kocan, Katherine M

    2007-01-31

    Anaplasma ovis (Rickettsiales: Anaplasmataceae) is a tick-borne pathogen of sheep, goats and wild ruminants. The genetic diversity of A. ovis strains has not been well characterized due to the lack of sequence information. In this study, we evaluated bighorn sheep (Ovis canadensis) and mule deer (Odocoileus hemionus) from Montana for infection with A. ovis by serology and sequence analysis of the msp4 gene. Antibodies to Anaplasma spp. were detected in 37% and 39% of bighorn sheep and mule deer analyzed, respectively. Four new msp4 genotypes were identified. The A. ovismsp4 sequences identified herein were analyzed together with sequences reported previously for the characterization of the genetic diversity of A. ovis strains in comparison with other Anaplasma spp. The results of these studies demonstrated that although A. ovismsp4 genotypes may vary among geographic regions and between sheep and deer hosts, the variation observed was less than the variation observed between A. marginale and A. phagocytophilum strains. The results reported herein further confirm that A. ovis infection occurs in natural wild ruminant populations in Western United States and that bighorn sheep and mule deer may serve as wildlife reservoirs of A. ovis.

  2. Sequence analysis of the msp4 gene of Anaplasma ovis strains

    USGS Publications Warehouse

    de la Fuente, J.; Atkinson, M.W.; Naranjo, V.; Fernandez de Mera, I. G.; Mangold, A.J.; Keating, K.A.; Kocan, K.M.

    2007-01-01

    Anaplasma ovis (Rickettsiales: Anaplasmataceae) is a tick-borne pathogen of sheep, goats and wild ruminants. The genetic diversity of A. ovis strains has not been well characterized due to the lack of sequence information. In this study, we evaluated bighorn sheep (Ovis canadensis) and mule deer (Odocoileus hemionus) from Montana for infection with A. ovis by serology and sequence analysis of the msp4 gene. Antibodies to Anaplasma spp. were detected in 37% and 39% of bighorn sheep and mule deer analyzed, respectively. Four new msp4 genotypes were identified. The A. ovis msp4 sequences identified herein were analyzed together with sequences reported previously for the characterization of the genetic diversity of A. ovis strains in comparison with other Anaplasma spp. The results of these studies demonstrated that although A. ovis msp4 genotypes may vary among geographic regions and between sheep and deer hosts, the variation observed was less than the variation observed between A. marginale and A. phagocytophilum strains. The results reported herein further confirm that A. ovis infection occurs in natural wild ruminant populations in Western United States and that bighorn sheep and mule deer may serve as wildlife reservoirs of A. ovis. ?? 2006.

  3. Differences in prevalence of Borrelia burgdorferi and Anaplasma spp. infection among host-seeking Dermacentor occidentalis, Ixodes pacificus, and Ornithodoros coriaceus ticks in northwestern California.

    PubMed

    Lane, Robert S; Mun, Jeomhee; Peribáñez, Miguel A; Fedorova, Natalia

    2010-12-01

    Previous studies revealed that the Pacific Coast tick (Dermacentor occidentalis) is infected occasionally with the agents of Lyme disease (Borrelia burgdorferi) or human granulocytic anaplasmosis (Anaplasma phagocytophilum) and that it is an inefficient experimental vector of B. burgdorferi. The relationship of the pajahuello tick (Ornithodoros coriaceus) to each of these bacterial zoonotic agents has not been reported. The primary bridging vector of both bacterial zoonotic agents to humans is the western black-legged tick (Ixodes pacificus). Because of the spatial and temporal overlap of D. occidentalis and O. coriaceus populations with those of I. pacificus in natural foci of B. burgdorferi and A. phagocytophilum in northwestern California, we conducted field and laboratory studies to determine if the Pacific Coast tick or the pajahuello tick potentially may serve as secondary vectors of either bacterium. Our findings reconfirmed that wild-caught D. occidentalis ticks are infected infrequently with B. burgdorferi or A. phagocytophilum, but some adult ticks from dense woodlands or chaparral were found to contain 2 important veterinary pathogens for the first time (Anaplasma bovis, A. ovis). The high prevalence of A. bovis infection (4.3%, n=185 ticks) within chaparral-derived ticks suggests that D. occidentalis could be an efficient vector of this rickettsia. Experimental attempts to transmit borreliae or Anaplasma spp. that may have been present in >100 wild-caught D. occidentalis adults to naïve rabbits were unsuccessful. Anaplasma spp. were not detected in O. coriaceus, but one (4.3%) of 23 nymphs was infected with B. bissettii. This finding and an antecedent report of a B. burgdorferi-like spirochete from the same tick species demonstrate that O. coriaceus sometimes acquires and transstadially passes Lyme disease group spirochetes. I. pacificus nymphs inhabiting a woodland nidus of B. burgdorferi and A. phagocytophilum had a 5-fold higher prevalence of

  4. Differences in prevalence of Borrelia burgdorferi and Anaplasma spp. infection among host-seeking Dermacentor occidentalis, Ixodes pacificus, and Ornithodoros coriaceus ticks in northwestern California

    PubMed Central

    Lane, Robert S.; Mun, Jeomhee; Peribáñez, Miguel A.; Fedorova, Natalia

    2010-01-01

    Previous studies revealed that the Pacific Coast tick (Dermacentor occidentalis) is infected occasionally with the agents of Lyme disease (Borrelia burgdorferi) or human granulocytic anaplasmosis (Anaplasma phagocytophilum) and that it is an inefficient experimental vector of B. burgdorferi. The relationship of the pajahuello tick (Ornithodoros coriaceus) to each of these bacterial zoonotic agents has not been reported. The primary bridging vector of both bacterial zoonotic agents to humans is the western black-legged tick (Ixodes pacificus). Because of the spatial and temporal overlap of D. occidentalis and O. coriaceus populations with those of I. pacificus in natural foci of B. burgdorferi and A. phagocytophilum in northwestern California, we conducted field and laboratory studies to determine if the Pacific Coast tick or the pajahuello tick potentially may serve as secondary vectors of either bacterium. Our findings reconfirmed that wild-caught D. occidentalis ticks are infected infrequently with B. burgdorferi or A. phagocytophilum, but some adult ticks from dense woodlands or chaparral were found to contain 2 important veterinary pathogens for the first time (Anaplasma bovis, A. ovis). The high prevalence of A. bovis infection (4.3%, n=185 ticks) within chaparral-derived ticks suggests that D. occidentalis could be an efficient vector of this rickettsia. Experimental attempts to transmit borreliae or Anaplasma spp. that may have been present in >100 wild-caught D. occidentalis adults to naïve rabbits were unsuccessful. Anaplasma spp. were not detected in O. coriaceus, but one (4.3%) of 23 nymphs was infected with B. bissettii. This finding and an antecedent report of a B. burgdorferi-like spirochete from the same tick species demonstrate that O. coriaceus sometimes acquires and transstadially passes Lyme disease group spirochetes. I. pacificus nymphs inhabiting a woodland nidus of B. burgdorferi and A. phagocytophilum had a 5-fold higher prevalence of

  5. Molecular Characterization of Anaplasma sp. in Free-Living Gray Brockets (Mazama gouazoubira).

    PubMed

    Mongruel, Anna Claudia Baumel; Benevenute, Jyan Lucas; André, Marcos Rogério; Carrasco, Adriano de Oliveira Torres; Machado, Rosangela Zacarias; Seki, Meire Christina

    2017-03-01

    Bacteria in the genus Anaplasma are responsible for diseases in animals and humans. Studies carried out in Brazil have demonstrated that Brazilian deer are able to act as hosts of agents in the family Anaplasmataceae and are possibly potential reservoirs of these pathogens. Molecular and phylogenetic studies have been carried out on samples of two gray brocket specimens (Mazama gouazoubira) from the city of Guarapuava, state of Paraná, Brazil, for the detection of Anaplasma sp. in these animals. Partial nucleotide sequences of the genes 16S rRNA and groESL were used for phylogenetic analyses and compared with other 13 and 17 partial sequences of the respective genes obtained in GenBank. These assessments showed topological incongruence among the trees generated in the phylogenetic analyses. Phylogenetic analysis based on the gene 16S rRNA of the genotypes amplified in the samples of this study was similar to those of A. bovis detected in dogs and wild deer in Japan, whereas studies carried out on gene groESL indicated proximity with sequences of Anaplasma sp. that were also isolated in deer in Japan and allocated in the same clade of partial sequences of A. phagocytophilum. As the 16S rRNA gene is highly conserved, with few polymorphic positions, it may show low reliability for studies on phylogenetic positioning. The present study detected an Anaplasma sp. genotype in two specimens of M. gouazoubira in southern Brazil, which may mean that this agent possibly circulates in deer populations, and demonstrated the need for studies related to the possible role of deer in enzootic cycles of Anaplasmataceae in Brazil.

  6. Molecular Detection of Zoonotic Rickettsiae and Anaplasma spp. in Domestic Dogs and Their Ectoparasites in Bushbuckridge, South Africa.

    PubMed

    Kolo, Agatha O; Sibeko-Matjila, Kgomotso P; Maina, Alice N; Richards, Allen L; Knobel, Darryn L; Matjila, Paul T

    2016-04-01

    Members of the order Rickettsiales are small, obligate intracellular bacteria that are vector-borne and can cause mild to fatal diseases in humans worldwide. There is little information on the zoonotic rickettsial pathogens that may be harbored by dogs from rural localities in South Africa. To characterize rickettsial pathogens infecting dogs, we screened 141 blood samples, 103 ticks, and 43 fleas collected from domestic dogs in Bushbuckridge Municipality, Mpumalanga Province of South Africa, between October 2011 and May 2012 using the reverse line blot (RLB) and Rickettsia genus and species-specific quantitative real-time PCR (qPCR) assays. Results from RLB showed that 49% of blood samples and 30% of tick pools were positive for the genus-specific probes for Ehrlichia/Anaplasma; 16% of the blood samples were positive for Ehrlichia canis. Hemoparasite DNA could not be detected in 36% of blood samples and 30% of tick pools screened. Seven (70%) tick pools and both flea pools were positive for Rickettsia spp; three (30%) tick pools were positive for Rickettsia africae; and both flea pools (100%) were positive for Rickettsia felis. Sequencing confirmed infection with R. africae and Candidatus Rickettsia asemboensis; an R. felis-like organism from one of the R. felis-positive flea pools. Anaplasma sp. South Africa dog strain (closely related to Anaplasma phagocytophilum), A. phagocytophilum, and an Orientia tsutsugamushi-like sequence were identified from blood samples. The detection of emerging zoonotic agents from domestic dogs and their ectoparasites in a rural community in South Africa highlights the potential risk of human infection that may occur with these pathogens.

  7. Spatio-temporal variations and genetic diversity of Anaplasma spp. in cattle from the North of Tunisia.

    PubMed

    Belkahia, Hanène; Ben Said, Mourad; El Mabrouk, Narjesse; Saidani, Mariem; Cherni, Chayma; Ben Hassen, Mariem; Bouattour, Ali; Messadi, Lilia

    2017-09-01

    In cattle, anaplasmosis is a tick-borne rickettsial disease caused by Anaplasma marginale, A. centrale, A. phagocytophilum, and A. bovis. To date, no information concerning the seasonal dynamics of single and/or mixed infections by different Anaplasma species in bovines are available in Tunisia. In this work, a total of 1035 blood bovine samples were collected in spring (n=367), summer (n=248), autumn (n=244) and winter (n=176) from five different governorates belonging to three bioclimatic zones from the North of Tunisia. Molecular survey of A. marginale, A. centrale and A. bovis in cattle showed that average prevalence rates were 4.7% (minimum 4.1% in autumn and maximum 5.6% in summer), 7% (minimum 3.9% in winter and maximum 10.7% in autumn) and 4.9% (minimum 2.7% in spring and maximum 7.3% in summer), respectively. A. phagocytophilum was not detected in all investigated cattle. Seasonal variations of Anaplasma spp. infection and co-infection rates in overall and/or according to each bioclimatic area were recorded. Molecular characterization of A. marginale msp4 gene indicated a high sequence homology of revealed strains with A. marginale sequences from African countries. Alignment of 16S rRNA A. centrale sequences showed that Tunisian strains were identical to the vaccine strain from several sub-Saharan African and European countries. The comparison of the 16S rRNA sequences of A. bovis variants showed a perfect homology between Tunisian variants isolated from cattle, goats and sheep. These present data are essential to estimate the risk of bovine anaplasmosis in order to develop integrated control policies against multi-species pathogen communities, infecting humans and different animal species, in the country. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Prevalence of Anaplasma, Bartonella and Borrelia Species in Haemaphysalis longicornis collected from goats in North Korea

    PubMed Central

    Kang, Jun-Gu; Ko, Sungjin; Smith, W. Barney; Kim, Heung-Chul; Lee, In-Yong

    2016-01-01

    North Korea is located on the northern part of the Korean Peninsula in East Asia. While tick-borne pathogens of medical and veterinary importance have been reported from China and South Korea, they have not been reported from North Korea. To screen for zoonotic tick-borne pathogens in North Korea, ticks were collected from domestic goats. A total of 292 (27 nymph, 26 male, 239 female) Haemaphysalis (H.) longicornis were collected and assayed individually for selected tick-borne pathogens. A total of 77 (26.4%) were positive for Anaplasma bovis, followed by Bartonella (B.) grahamii (15, 5.1%), Anaplasma phagocytophilum (12, 4.1%), Bartonella henselae (10, 3.4%), and Borrelia spp. (3, 1.0%) based on 16S ribosomal RNA and ITS species-specific nested polymerase chain reaction. Using the groEL-based nested PCR, a total of 6 and 1 H. longicornis were positive for B. grahamii and B. henselae, respectively. All products were sequenced and demonstrated 100% identity and homology with previously reported sequences from other countries in GenBank. This is the first report of the detection of tick-borne pathogens in the North Korea and suggests that farm animals may act as reservoirs for zoonotic tick-borne pathogens. PMID:26645342

  9. Prevalence of Anaplasma, Bartonella and Borrelia Species in Haemaphysalis longicornis collected from goats in North Korea.

    PubMed

    Kang, Jun-Gu; Ko, Sungjin; Smith, W Barney; Kim, Heung-Chul; Lee, In-Yong; Chae, Joon-Seok

    2016-06-30

    North Korea is located on the northern part of the Korean Peninsula in East Asia. While tick-borne pathogens of medical and veterinary importance have been reported from China and South Korea, they have not been reported from North Korea. To screen for zoonotic tick-borne pathogens in North Korea, ticks were collected from domestic goats. A total of 292 (27 nymph, 26 male, 239 female) Haemaphysalis (H.) longicornis were collected and assayed individually for selected tick-borne pathogens. A total of 77 (26.4%) were positive for Anaplasma bovis, followed by Bartonella (B.) grahamii (15, 5.1%), Anaplasma phagocytophilum (12, 4.1%), Bartonella henselae (10, 3.4%), and Borrelia spp. (3, 1.0%) based on 16S ribosomal RNA and ITS species-specific nested polymerase chain reaction. Using the groEL-based nested PCR, a total of 6 and 1 H. longicornis were positive for B. grahamii and B. henselae, respectively. All products were sequenced and demonstrated 100% identity and homology with previously reported sequences from other countries in GenBank. This is the first report of the detection of tick-borne pathogens in the North Korea and suggests that farm animals may act as reservoirs for zoonotic tick-borne pathogens.

  10. Investigation of tick-borne bacteria (Rickettsia spp., Anaplasma spp., Ehrlichia spp. and Borrelia spp.) in ticks collected from Andean tapirs, cattle and vegetation from a protected area in Ecuador.

    PubMed

    Pesquera, Cristina; Portillo, Aránzazu; Palomar, Ana M; Oteo, José A

    2015-01-24

    Ixodid ticks play an important role in the transmission and ecology of infectious diseases. Information about the circulation of tick-borne bacteria in ticks is lacking in Ecuador. Our aims were to investigate the tick species that parasitize Andean tapirs and cattle, and those present in the vegetation from the buffer zone of the Antisana Ecological Reserve and Cayambe-Coca National Park (Ecuador), and to investigate the presence of tick-borne bacteria. Tick species were identified based on morphologic and genetic criteria. Detection of tick-borne bacteria belonging to Rickettsia, Anaplasma, Ehrlichia and Borrelia genera was performed by PCRs. Our ticks included 91 Amblyomma multipunctum, 4 Amblyomma spp., 60 Rhipicephalus microplus, 5 Ixodes spp. and 1 Ixodes boliviensis. A potential Candidatus Rickettsia species closest to Rickettsia monacensis and Rickettsia tamurae (designated Rickettsia sp. 12G1) was detected in 3 R. microplus (3/57, 5.3%). In addition, Anaplasma spp., assigned at least to Anaplasma phagocytophilum (or closely related genotypes) and Anaplasma marginale, were found in 2 A. multipunctum (2/87, 2.3%) and 13 R. microplus (13/57, 22.8%). This is the first description of Rickettsia sp. in ticks from Ecuador, and the analyses of sequences suggest the presence of a potential novel Rickettsia species. Ecuadorian ticks from Andear tapirs, cattle and vegetation belonging to Amblyomma and Rhipicephalus genera were infected with Anaplasmataceae. Ehrlichia spp. and Borrelia burgdorferi sensu lato were not found in any ticks.

  11. Molecular characterization and specific detection of Anaplasma species (AP-sd) in sika deer and its first detection in wild brown bears and rodents in Hokkaido, Japan.

    PubMed

    Moustafa, Mohamed Abdallah Mohamed; Lee, Kyunglee; Taylor, Kyle; Nakao, Ryo; Sashika, Mariko; Shimozuru, Michito; Tsubota, Toshio

    2015-12-01

    A previously undescribed Anaplasma species (herein referred to as AP-sd) has been detected in sika deer, cattle and ticks in Japan. Despite being highly similar to some strains of A. phagocytophilum, AP-sd has never been detected in humans. Its ambiguous epidemiology and the lack of tools for its specific detection make it difficult to understand and interpret the prevalence of this Anaplasma species. We developed a method for specific detection, and examined AP-sd prevalence in Hokkaido wildlife. Our study included 250 sika deer (Cervus nippon yesoensis), 13 brown bears (Ursus arctos yesoensis) and 252 rodents including 138 (Apodemus speciosus), 45 (Apodemus argenteus), 42 (Myodes rufocanus) and 27 (Myodes rutilus) were collected from Hokkaido island, northern Japan, collected during 2010 to 2015. A 770 bp and 382 bp segment of the 16S rRNA and gltA genes, respectively, were amplified by nested PCR. Results were confirmed by cloning and sequencing of the positive PCR products. A reverse line blot hybridization (RLB) based on the 16S rRNA gene was then developed for the specific detection of AP-sd. The prevalence of AP-sd by nested PCR in sika deer was 51% (128/250). We detected this Anaplasma sp. for the first time in wild brown bears and rodents with a prevalence of 15% (2/13) and 2.4% (6/252), respectively. The sequencing results of the 16S rRNA and gltA gene amplicons were divergent from the selected A. phagocytophilum sequences in GenBank. Using a newly designed AP-sd specific probe for RLB has enabled us to specifically detect this Anaplasma species. Besides sika deer and cattle, wild brown bears and rodents were identified as potential reservoir hosts for AP-sd. This study provided a high throughput molecular method that specifically detects AP-sd, and which can be used to investigate its ecology and its potential as a threat to humans in Japan.

  12. Larvae of chigger mites Neotrombicula spp. (Acari: Trombiculidae) exhibited Borrelia but no Anaplasma infections: a field study including birds from the Czech Carpathians as hosts of chiggers.

    PubMed

    Literak, Ivan; Stekolnikov, Alexandr A; Sychra, Oldrich; Dubska, Lenka; Taragelova, Veronika

    2008-04-01

    Chigger mites were collected from 1,080 wild birds of 37 species at Certak (Czech Republic), in the western Carpathian Mountains, from 29 July to 24 September 2005. The prevalence of infestation with chigger larvae was 7%. A total of 325 chigger specimens from 10 bird species was identified and three chigger species were found: Neotrombicula autumnalis, N. carpathica, and N. inopinata, the latter two species being reported on new hosts. Neotrombicula carpathica is reported in the Czech Republic for the first time. A total of 509 chigger larvae found on 79 host specimens were examined by polymerase chain reaction (PCR) for the presence of Borrelia burgdorferi s.l. DNA (fragments of the rrf (5S)--rrl (23S) intergenic spacer), and Anaplasma phagocytophilum DNA (epank1 gene). A fragment of specific Borrelia DNA was amplified through PCR in one sample, and the PCR product was further analyzed by reverse line blotting assay, whereby both genospecies of B. garinii and B. valaisiana were proved. This sample pooled five chigger larvae collected from one Sylvia atricapilla on 11 August 2005. No A. phagocytophilum DNA was amplified. We conclude that larvae of the genus Neotrombicula can be infected with Borrelia genospecies originated from their present or former hosts.

  13. Expression of Anaplasma marginale ankyrin repeat-containing proteins during infection of the mammalian host and tick vector.

    PubMed

    Ramabu, Solomon S; Schneider, David A; Brayton, Kelly A; Ueti, Massaro W; Graça, Telmo; Futse, James E; Noh, Susan M; Baszler, Timothy V; Palmer, Guy H

    2011-07-01

    Transmission of tick-borne pathogens requires transition between distinct host environments with infection and replication in host-specific cell types. Anaplasma marginale illustrates this transition: in the mammalian host, the bacterium infects and replicates in mature (nonnucleated) erythrocytes, while in the tick vector, replication occurs in nucleated epithelial cells. We hypothesized that proteins containing ankyrin motifs would be expressed by A. marginale only in tick cells and would traffic to the infected host cell nucleus. A. marginale encodes three proteins containing ankyrin motifs, an AnkA orthologue (the AM705 protein), AnkB (the AM926 protein), and AnkC (the AM638 protein). All three A. marginale Anks were confirmed to be expressed during intracellular infection: AnkA is expressed at significantly higher levels in erythrocytes, AnkB is expressed equally by both infected erythrocytes and tick cells, and AnkC is expressed exclusively in tick cells. There was no evidence of any of the Ank proteins trafficking to the nucleus. Thus, the hypothesis that ankyrin-containing motifs were predictive of cell type expression and nuclear localization was rejected. In contrast, AnkA orthologues in the closely related A. phagocytophilum and Ehrlichia chaffeensis have been shown to localize to the host cell nucleus. This difference, together with the lack of a nuclear localization signal in any of the AnkA orthologues, suggests that trafficking may be mediated by a separate transporter rather than by endogenous signals. Selection for divergence in Ank function among Anaplasma and Ehrlichia spp. is supported by both locus and allelic analyses of genes encoding orthologous proteins and their ankyrin motif compositions.

  14. Molecular survey and characterization of a novel Anaplasma species closely related to Anaplasma capra in ticks, northwestern China.

    PubMed

    Yang, Jifei; Liu, Zhijie; Niu, Qingli; Liu, Junlong; Han, Rong; Liu, Guangyuan; Shi, Yaoxu; Luo, Jianxun; Yin, Hong

    2016-11-25

    Anaplasma spp. are tick-transmitted bacteria that infect a wide variety of wild and domestic animals. These pathogens exhibit a high degree of biological diversity, broad geographical distribution, and represent a serious threat to veterinary and public health worldwide. A novel Anaplasma species was identified in Haemaphysalis qinghaiensis (Ixodidae) in northwestern China and was molecularly characterized by comparison of 16S rRNA, gltA, and groEL gene sequences. Of the 414 samples tested, 24 (5.8%) were positive for this Anaplasma species. On the basis of the 16S rRNA gene, this organism has been found to be closely related to and exhibit the highest sequence similarity with A. capra (99.8-99.9%) that was identified in goats and humans in northern China, but was distinct from other known Anaplasma species. Sequence analysis of the gltA and groEL genes revealed that this Anaplasma species was distinct from A. capra considering the lower sequence identity (88.6-88.7% for gltA and 90.6-91.0% for groEL) and a divergent phylogenetic position. Therefore, we described this Anaplasma species as A. capra-like bacteria. The present study reports a potential novel Anaplasma species closely related to A. capra in H. qinghaiensis in northwestern China. The zoonotic potential of A. capra-like bacteria needs to be further determined.

  15. Detection of Rickettsia and Anaplasma from hard ticks in Thailand.

    PubMed

    Malaisri, Premnika; Hirunkanokpun, Supanee; Baimai, Visut; Trinachartvanit, Wachareeporn; Ahantarig, Arunee

    2015-12-01

    We collected a total of 169 adult hard ticks and 120 nymphs from under the leaves of plants located along tourist nature trails in ten localities. The results present data examining the vector competence of ticks of different genera and the presence of Rickettsia and Anaplasma species. The ticks belonged to three genera, Amblyomma, Dermacentor, and Haemaphysalis, comprising 11 species. Rickettsia bacteria were detected at three collection sites, while Anaplasma bacteria were detected at only one site. Phylogenetic analysis revealed new rickettsia genotypes from Thailand that were closely related to Rickettsia tamurae, Rickettsia monacensis, and Rickettsia montana. This study was also the first to show that Anaplasma bacteria are found in Haemaphysalis shimoga ticks and are closely related evolutionarily to Anaplasma bovis. These results provide additional information for the geographical distribution of tick species and tick-borne bacteria in Thailand and can therefore be applied for ecotourism management.

  16. Molecular detection of Anaplasma platys, Ehrlichia canis, Hepatozoon canis and Rickettsia monacensis in dogs from Maio Island of Cape Verde archipelago.

    PubMed

    Lauzi, Stefania; Maia, João P; Epis, Sara; Marcos, Ricardo; Pereira, Cristina; Luzzago, Camilla; Santos, Marta; Puente-Payo, Pablo; Giordano, Alessia; Pajoro, Massimo; Sironi, Giuseppe; Faustino, Augusto

    2016-07-01

    Tick-borne diseases are emerging worldwide and have an important zoonotic relevance. Dogs play an important role in the epidemiology of several zoonotic tick-borne pathogens acting as sentinels and/or reservoirs. This study focused on the molecular identification of tick-borne pathogens in blood samples of 153 autochthonous asymptomatic dogs in Maio Island, Cape Verde archipelago. Eighty-four (54.9%) dogs were positive for one or more pathogens. Fifty-five (35.9%) dogs were infected with Hepatozoon canis, 53 (34.6%) with Anaplasma platys, five (3.3%) with Ehrlichia canis and Rickettsia monacensis, an emerging human pathogen, was also identified in a single dog (0.7%). The former three pathogens cause important canine tick-borne diseases that are transmitted or potentially transmitted by Rhipicephalus sanguineus s.l., the only hard tick identified in Cape Verde. Furthermore, Wolbachia spp. was amplified from the blood of one dog. None of the dogs were positive for Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Midichloria mitochondrii, Bartonella spp., Babesia spp. or Theileria spp. Fifty-four (35.3%) animals showed single infections and 30 (19.6%) co-infections, with A. platys and H. canis co-infection being the most frequent (28 dogs, 18.3%). The frequency of E. canis infection was statistically different among age groups (P=0.017), being higher among dogs older than 4 years compared to younger dogs. Infection by A. platys was also statistically different among age groups (P=0.031), being higher in dogs younger than 2 years compared to older dogs. The statistical analyses showed no significant association of PCR positivity with gender or location. The frequency of tick-borne pathogens detected in dogs in Maio Island, including R. monacensis, highlights the need to improve diagnosis and control in order to prevent the risk of transmission of these pathogens among dogs and humans living in or travelling to this touristic island.

  17. Recrudescence of Anaplasma marginale induced by immunosuppression with cyclophosphamide.

    PubMed

    Corrier, D E; Wagner, G G; Adams, L G

    1981-01-01

    Eight doses of cyclophosphamide (5 mg/kg), at 2-day intervals between doses, were administered IV to Anaplasma-carrier splenectomized calves. Significant decreases in serum immunoglobulins, diminished complement-fixing antibody response, and transitory leukopenia were associated with the treatment. Recrudescence of clinical Anaplasma infection occurred after the 5th dose of cyclophosphate was given and was characterized by rapid increase in parasitemia, marked decrease in PCV, and mortality. The results of the present study indicated that humoral mediated immunity may contribute to the maintenance of a state of equilibrium in the host-parasite relationship and that suppression of humoral immune responses may alter the course and outcome of infection in Anaplasma carriers.

  18. Seroprevalence of Anaplasma marginale in Texas cattle.

    PubMed

    Hairgrove, Thomas B; Craig, Thomas M; Budke, Christine M; Rodgers, Sandy J; Gill, Ronald J

    2014-09-01

    To our knowledge the seroprevalence of Anaplasma marginale in Texas has not been reported. The objective of this study was to estimate the point seroprevalence and spatial distribution of Texas cattle persistently infected with A. marginale. This was a cross-sectional observational study examining serum collected from 12,000 adult cattle marketed in 23 selected Texas auction markets during the second week of July 2011. A random subset of those cattle comprising 1835 individuals was evaluated for persistent infection with A. marginale using a commercial cELISA for antibody detection. The pooled apparent seroprevalence for cattle tested at auction markets across the state was 15.02% (95% CI: 11.02-19.53%), with markets in the western portion of the state demonstrating prevalence ⇒ 30%. The winter tick, Dermacentor albipictus is involved in the biological transfer of A. marginale and is prevalent in west Texas. Producers in endemic and non-endemic areas should be encouraged to determine the infection status of replacement cattle in order to implement effective management strategies for the control bovine anaplasmosis. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Anaplasma platys in dogs from Uruguay.

    PubMed

    Carvalho, Luis; Armua-Fernandez, Maria Teresa; Sosa, Nicolás; Félix, María Laura; Venzal, José Manuel

    2017-02-01

    Anaplasmataceae family members include vector-borne bacteria of veterinary importance that may also affect humans. Ehrlichia canis and Anaplasma platys are the main members of this family detected in dogs worldwide. In Uruguay there are not many published studies on tick-borne pathogens affecting dogs, the only haemoparasite molecularly confirmed in dogs, is the piroplasm Rangelia vitalii. The aim of the present work was to detect the presence of A. platys and E. canis in dogs and dogs-associated ticks of two localities in Northwestern Uruguay. Blood samples from dogs with and without clinical signs associated with vector-borne diseases, and Rhipicephalus sanguineus obtained from these dogs were analyzed by PCR for Anaplasmataceae. Positive dogs were further analyzed by PCR for Ehrlichia spp. and A. platys. All the ticks were found negative. No dog was detected infected with E. canis, while eight dogs (4.2%) were found to be infected with A. platys. Phylogenetic analysis of groESL operon sequence for A. platys revealed no differences with sequences described for A. platys in neighbor countries and from other regions of the world. This is the first report of the presence of A. platys in Uruguay.

  20. Experimental infections of Anaplasma ovis in pronghorn antelope.

    PubMed

    Zaugg, J L

    1987-04-01

    Anaplasma ovis was experimentally transmitted from sheep to pronghorn antelope (Antilocapra americana) and back to sheep. Anaplasma ovis was recovered in splenectomized sheep, from two of three spleen-intact pronghorns following their inoculation with blood from known A. ovis carrier sheep. These two pronghorns exhibited a 0.5% or higher A. ovis parasitemia within 48 days after exposure, and an anaplasmosis-positive serological response 91 days after exposure. Clinical signs of illness were not observed. Blood from the infected pronghorns produced disease in four splenectomized sheep.

  1. Anaplasma platys in Bone Marrow Megakaryocytes of Young Dogs

    PubMed Central

    De Tommasi, A. Sara; Baneth, Gad; Breitschwerdt, Edward B.; Stanneck, Dorothee; Dantas-Torres, Filipe; Otranto, Domenico

    2014-01-01

    Anaplasma platys is an obligate intracellular rickettsial pathogen that infects platelets of dogs, forming basophilic intracellular morulae. In the present report, cellular inclusions were documented in bone marrow thrombocyte precursors of two young naturally infected dogs, indicating that A. platys can infect megakaryocytes and promegakaryocytes. PMID:24622106

  2. Detection of Anaplasma sp. in Korean Native Goats (Capra aegagrus hircus) on Jeju Island, Korea

    PubMed Central

    Seong, Giyong; Han, Yu-Jung; Chae, Jeong-Byoung; Chae, Joon-Seok; Yu, Do-Hyeon; Lee, Young-Sung; Park, Jinho; Park, Bae-Keun; Yoo, Jae-Gyu; Choi, Kyoung-Seong

    2015-01-01

    Anaplasma species are obligate intracellular pathogens that can cause tick-borne diseases in mammalian hosts. To date, very few studies of their occurrence in Korean native goats (Capra aegagrus hircus) have been reported. In the present study, we investigated Anaplasma infection of Korean native goats on Jeju Island, Republic of Korea, and performed phylogenetic analysis based on the 16S rRNA gene sequences. Our results showed that Anaplasma infection was found mostly in adult female goats. The phylogenetic tree revealed that the 7 sequences identified in Korean native goats could belong to Anaplasma sp. and were distinct from A. marginale, A. centrale, and A. ovis. The results indicated that the sequences identified to belong to Anaplasma were closely related to sequences isolated from goats in China and were clustered within the same group. To our knowledge, this is the first study to detect Anaplasma sp. infection in Korean native goats. PMID:26797447

  3. Detection of Anaplasma sp. in Korean Native Goats (Capra aegagrus hircus) on Jeju Island, Korea.

    PubMed

    Seong, Giyong; Han, Yu-Jung; Chae, Jeong-Byoung; Chae, Joon-Seok; Yu, Do-Hyeon; Lee, Young-Sung; Park, Jinho; Park, Bae-Keun; Yoo, Jae-Gyu; Choi, Kyoung-Seong

    2015-12-01

    Anaplasma species are obligate intracellular pathogens that can cause tick-borne diseases in mammalian hosts. To date, very few studies of their occurrence in Korean native goats (Capra aegagrus hircus) have been reported. In the present study, we investigated Anaplasma infection of Korean native goats on Jeju Island, Republic of Korea, and performed phylogenetic analysis based on the 16S rRNA gene sequences. Our results showed that Anaplasma infection was found mostly in adult female goats. The phylogenetic tree revealed that the 7 sequences identified in Korean native goats could belong to Anaplasma sp. and were distinct from A. marginale, A. centrale, and A. ovis. The results indicated that the sequences identified to belong to Anaplasma were closely related to sequences isolated from goats in China and were clustered within the same group. To our knowledge, this is the first study to detect Anaplasma sp. infection in Korean native goats.

  4. Anaplasma sp. and hemoplasma infection in leopard cats (Prionailurus bengalensis euptilurus) from Korea

    PubMed Central

    Hwang, Jusun; Oh, Dae-Hyun; Lee, Hang

    2015-01-01

    This study examined the occurrence of Anaplasma spp. and hemoplasma infection in leopard cats, Prionailurus bengalensis euptilurus, in Korea. Twenty-nine biological samples were tested by molecular analysis. Two (6.9%) and eight (27.6%) tested specimens were positive for Anaplasma bovis and hemoplasma infection, respectively. Based on our results, Anaplasma/Ehrlichia spp. and hemoplasma are regularly infecting leopard cat populations of Korea. Considering their endangered status, regular monitoring of infection by arthropod-borne pathogens known to cause clinical symptoms in feline hosts such as Anaplasma/Ehrlichia spp. and hemoplasma would be crucial as part of ongoing conservation efforts. PMID:26040618

  5. Knockdown of the rhipicephalus microplus cytochrome c oxidase subunit III gene is associated with a failure of anaplasma marginale transmission

    USDA-ARS?s Scientific Manuscript database

    Rhipicephalus microplus is an obligate hematophagous ectoparasite of cattle and an important biological vector of Anaplasma marginale in tropical and subtropical regions. The primary determinants for Anaplasma transmission are infection of tick gut epithelial cells followed by infection of salivary ...

  6. Evaluation of jackrabbits as nonruminant hosts for Anaplasma marginale.

    PubMed

    Zaugg, J L; Newman, B A

    1985-03-01

    Two black-tailed jackrabbits (Lepus californicus), 1 splenectomized and 1 intact, were inoculated with 0.2 ml of a 1:5 dilution of a Florida Anaplasma marginale stabilate. Five months later, both hares were inoculated with 1 ml of whole blood from a calf with acute anaplasmosis. Neither hare developed any signs of clinical anaplasmosis. Pooled blood (7 ml) from these jackrabbits which was inoculated into 2 Anaplasma-susceptible, splenectomized calves failed to induce hematologic or serologic signs of anaplasmosis for at least 90 days. Two susceptible, splenectomized calves were inoculated with 35 ml of pooled whole blood from 9 wild-collected black-tailed jackrabbits from a known anaplasmosis enzootic area. Both steers remained free of anaplasmosis signs for 90 days.

  7. Anaplasma marginale and Theileria annulata in questing ticks from Portugal.

    PubMed

    Antunes, S; Ferrolho, J; Domingues, N; Santos, A S; Santos-Silva, M M; Domingos, A

    2016-09-01

    Ticks are ubiquitous arthropods and vectors of several pathogenic agents in animals and humans. Monitoring questing ticks is of great importance to ascertain the occurrence of pathogens and the potential vector species, offering an insight into the risk of disease transmission in a given area. In this study 428 host-seeking ticks, belonging to nine species of Ixodidae and collected from 17 of the 23 Portuguese mainland subregions, were screened for several tick-borne agents with veterinary relevance: Anaplasma marginale, Anaplasma ovis, Anaplasma centrale, Babesia spp., Coxiella burnetii and Theileria spp. Prevalence was assessed by PCR and amplified amplicons sequenced for validation of results. Twenty ticks, in a total of 428, were found positive: one Ixodes ventalloi for Theileria annulata and four Dermacentor marginatus, one Haemaphysalis punctata, five Ixodes ricinus, five I. ventalloi, and four Rhipicephalus sanguineus sensu lato for A. marginale. According to the reviewed literature, this is the first report of A. marginale and T. annulata detection in I. ventalloi. Furthermore, the amplification of A. marginale DNA in several tick species suggests a broad range for this agent in Portugal that might include other uncommon species as R. sanguineus s.l. This work provides new data towards a better understanding of tick-pathogen associations and also contributes to the surveillance of tick-borne agents in geographic areas with limited information.

  8. Anaplasma odocoilei sp. nov. (family Anaplasmataceae) from white-tailed deer (Odocoileus virginianus).

    PubMed

    Tate, Cynthia M; Howerth, Elizabeth W; Mead, Daniel G; Dugan, Vivien G; Luttrell, M Page; Sahora, Alexandra I; Munderloh, Ulrike G; Davidson, William R; Yabsley, Michael J

    2013-02-01

    Recently, an undescribed Anaplasma sp. (also called Ehrlichia-like sp. or WTD agent) was isolated in ISE6 tick cells from captive white-tailed deer. The goal of the current study was to characterize this organism using a combination of experimental infection, morphologic, serologic, and molecular studies. Each of 6 experimentally inoculated white-tailed deer fawns (Odocoileus virginianus) became chronically infected (100+ days) with the Anaplasma sp. by inoculation of either infected whole blood or culture. None of the deer showed evidence of clinical disease, but 3 of the 6 deer evaluated had multiple episodes of transient thrombocytopenia. Light microscopy of Giemsa-stained, thin blood smears revealed tiny, dark, spherical structures in platelets of acutely infected deer. Anaplasma sp. was detected in platelets of inoculated deer by polymerase chain reaction, transmission electron microscopy, immunohistochemistry, and in situ hybridization. Five of 6 deer developed antibodies reactive to Anaplasma sp. antigen, as detected by indirect fluorescent antibody testing. Phylogenetic analyses of 16S rRNA, groESL, and gltA sequences confirmed the Anaplasma sp. is related to A. platys. Two attempts to transmit the Anaplasma sp. between deer by feeding Amblyomma americanum, a suspected tick vector, were unsuccessful. Based on its biologic, antigenic, and genetic characteristics, this organism is considered a novel species of Anaplasma, and the name Anaplasma odocoilei sp. nov. is proposed with UMUM76(T) (=CSUR-A1) as the type strain. Copyright © 2012 Elsevier GmbH. All rights reserved.

  9. Anaplasma odocoilei sp. nov. (family Anaplasmataceae) from white-tailed deer (Odocoileus virginianus)

    PubMed Central

    Tate, Cynthia M.; Howerth, Elizabeth W.; Mead, Daniel G.; Dugan, Vivien G.; Luttrell, M. Page; Sahora, Alexandra I.; Munderloh, Ulrike G.; Davidson, William R.; Yabsley, Michael J.

    2014-01-01

    Recently, an undescribed Anaplasma sp. (also called Ehrlichia-like sp. or WTD agent) was isolated in ISE6 tick cells from captive white-tailed deer. The goal of the current study was to characterize this organism using a combination of experimental infection, morphologic, serologic, and molecular studies. Each of 6 experimentally inoculated white-tailed deer fawns (Odocoileus virginianus) became chronically infected (100+ days) with the Anaplasma sp. by inoculation of either infected whole blood or culture. None of the deer showed evidence of clinical disease, but 3 of the 6 deer evaluated had multiple episodes of transient thrombocytopenia. Light microscopy of Giemsa-stained, thin blood smears revealed tiny, dark, spherical structures in platelets of acutely infected deer. Anaplasma sp. was detected in platelets of inoculated deer by polymerase chain reaction, transmission electron microscopy, immunohistochemistry, and in situ hybridization. Five of 6 deer developed antibodies reactive to Anaplasma sp. antigen, as detected by indirect fluorescent antibody testing. Phylogenetic analyses of 16S rRNA, groESL, and gltA sequences confirmed the Anaplasma sp. is related to A. platys. Two attempts to transmit the Anaplasma sp. between deer by feeding Amblyomma americanum, a suspected tick vector, were unsuccessful. Based on its biologic, antigenic, and genetic characteristics, this organism is considered a novel species of Anaplasma, and the name Anaplasma odocoilei sp. nov. is proposed with UMUM76T (=CSUR-A1) as the type strain. PMID:23276749

  10. Validation of an improved Anaplasma antibody competitive ELISA for detection of Anaplasma ovis antibody in domestic sheep.

    PubMed

    Mason, Kathleen L; Gonzalez, Michael V; Chung, Chungwon; Mousel, Michelle R; White, Stephen N; Taylor, Joshua B; Scoles, Glen A

    2017-09-01

    An accurate and simple-to-perform new version of a competitive ELISA (cELISA) kit that became commercially available in 2015 for testing of cattle for antibody to Anaplasma marginale was validated for detection of Anaplasma ovis antibody in domestic sheep. True positives and negatives were identified using nested PCR (nPCR) as the gold standard. Negative bovine control sera supplied with the kit were used to calculate % inhibition (%I), designated bovine control ELISA (BcELISA), and this was compared to %I calculated from negative ovine sera derived from hand-raised, pathogen-free sheep, designated ovine control ELISA (OcELISA). The receiver operating characteristics area under the curve was 1.0 with a p value <0.001 regardless of the source of the control sera. The cutoff values for negative BcELISA and OcELISA were <30%I and <27%I, respectively. Our work confirmed that this Anaplasma antibody cELISA kit version 2 can be used with the serum controls supplied in the kit to test for A. ovis antibody in domestic sheep. Furthermore, this work confirmed the historically high infection prevalence (>93%) at the U.S. Sheep Experiment Station (Dubois, Idaho), in spite of efforts to reduce the possibility for iatrogenic transmission there, suggesting high levels of tick-borne transmission.

  11. Anaplasma marginale Outer Membrane Protein A Is an Adhesin That Recognizes Sialylated and Fucosylated Glycans and Functionally Depends on an Essential Binding Domain.

    PubMed

    Hebert, Kathryn S; Seidman, David; Oki, Aminat T; Izac, Jerilyn; Emani, Sarvani; Oliver, Lee D; Miller, Daniel P; Tegels, Brittney K; Kannagi, Reiji; Marconi, Richard T; Carlyon, Jason A

    2017-03-01

    Anaplasma marginale causes bovine anaplasmosis, a debilitating and potentially fatal tick-borne infection of cattle. Because A. marginale is an obligate intracellular organism, its adhesins that mediate entry into host cells are essential for survival. Here, we demonstrate that A. marginale outer membrane protein A (AmOmpA; AM854) contributes to the invasion of mammalian and tick host cells. AmOmpA exhibits predicted structural homology to OmpA of A. phagocytophilum (ApOmpA), an adhesin that uses key lysine and glycine residues to interact with α2,3-sialylated and α1,3-fucosylated glycan receptors, including 6-sulfo-sialyl Lewis x (6-sulfo-sLe(x)). Antisera against AmOmpA or its predicted binding domain inhibits A. marginale infection of host cells. Residues G55 and K58 are contributory, and K59 is essential for recombinant AmOmpA to bind to host cells. Enzymatic removal of α2,3-sialic acid and α1,3-fucose residues from host cell surfaces makes them less supportive of AmOmpA binding. AmOmpA is both an adhesin and an invasin, as coating inert beads with it confers adhesiveness and invasiveness. Recombinant forms of AmOmpA and ApOmpA competitively antagonize A. marginale infection of host cells, but a monoclonal antibody against 6-sulfo-sLe(x) fails to inhibit AmOmpA adhesion and A. marginale infection. Thus, the two OmpA proteins bind related but structurally distinct receptors. This study provides a detailed understanding of AmOmpA function, identifies its essential residues that can be targeted by blocking antibody to reduce infection, and determines that it binds to one or more α2,3-sialylated and α1,3-fucosylated glycan receptors that are unique from those targeted by ApOmpA.

  12. Molecular Survey of Anaplasma and Ehrlichia of Red Deer and Sika Deer in Gansu, China in 2013.

    PubMed

    Li, Y; Chen, Z; Liu, Z; Liu, J; Yang, J; Li, Q; Li, Y; Luo, J; Yin, H

    2016-12-01

    Anaplasma and Ehrlichia are important emerging tick-borne pathogens in both humans and animals. Here, we conducted a molecular surveillance study in Gansu, China to assess the prevalence of Anaplasma and Ehrlichia spp. in red deer and sika deer based on polymerase chain reaction (PCR) analysis and sequencing of 16S rRNA or msp genes. PCR revealed that the prevalence of Anaplasma ovis, Anaplasma bovis and Anaplasma platys of the Qilian Mountain samples was 32%, 9% and 9%, respectively; the prevalence of Anaplasma ovis, Anaplasma bovis, Anaplasma platys was 20%, 15% and 15% among the Long Mountain samples, respectively. Of the Long Mountain samples, two (5%) of the 40 samples were positive for Ehrlichia canis, but all 44 of the Qilian Mountain samples were negative for E. canis, and no other Anaplasma or Ehrlichia spp. were found in the samples. The phylogenetic tree showed that the newly isolated Anaplasma and Ehrlichia spp. could be classified as belonging to four clades, including an A. bovis cluster, A. ovis cluster, A. platys cluster and E. canis cluster. In addition, Bartonella schoenbuchensis was firstly identified in blood samples from red deer in Gansu, China. Our results provide important data to increase the understanding of the epidemiology of anaplasmosis and ehrlichiosis of red deer and sika deer and will assist with the implementation of measures to control anaplasmosis and ehrlichiosis transmission to red deer, sika deer and other animals in Gansu, China.

  13. Acute phase response in cattle infected with Anaplasma marginale.

    PubMed

    Nazifi, S; Razavi, S M; Kaviani, F; Rakhshandehroo, E

    2012-03-23

    This study was undertaken to evaluate the acute phase responses via the assessment of the concentration of serum sialic acids (total, lipid bound and protein bound), inflammatory mediators (IFN-γ and TNF-α) and acute phase proteins (Hp and SAA) in 20 adult crossbred cattle naturally infected by Anaplasma marginale. The infected animals were divided into 2 subgroups on the basis of parasitemia rate (<20% and >20%). Also, as a control group, 10 clinically healthy cattle from the same farms were sampled. Our data revealed significant decreases in red blood cell count (RBC), hematocrite (PCV) and hemoglobine (Hb) in infected cattle compared to healthy ones. Conversely, the concentrations of Hp, SAA, ceruloplasmin, fibrinogen, serum sialic acids and the circulatory IFN-γ and TNF-α were increased in the diseased cattle (P<0.05). In addition, it was evident that the progression of parasitemia in infected cattle did not induce any significant alterations in the hematological indices (RBCs, PCV and Hb) and the concentrations of Hp, SAA, ceruloplasmin and fibrinogen. SAA was the most sensitive factor to change in the diseased cattle. Therefore, increase in SAA concentration may be a good indicator of inflammatory process in cattle naturally infected with Anaplasma marginale.

  14. Molecular detection of Ehrlichia canis, Anaplasma bovis, Anaplasma platys, Candidatus Midichloria mitochondrii and Babesia canis vogeli in ticks from Israel.

    PubMed

    Harrus, S; Perlman-Avrahami, A; Mumcuoglu, K Y; Morick, D; Eyal, O; Baneth, G

    2011-03-01

    : Ticks are vectors of important pathogens of human and animals. Therefore, their microbial carriage capacity is constantly being investigated. The aim of this study was to characterize the diversity of domestic animal pathogens in ticks collected from vegetation and the ground, from different parts of Israel. Non-engorged questing adult ticks were collected from 13 localities. A total of 1196 ticks in 131 pools-83 pools of Rhipicephalus turanicus and 48 of Rhipicephalus sanguineus (with two to ten ticks per pool)-were included in this study. In addition, 13 single free-roaming Hyalomma spp. ticks were collected. Screening by molecular techniques revealed the presence of Ehrlichia canis, Anaplasma platys, Anaplasma bovis and Babesia canis vogeli DNA in R. turanicus ticks. E. canis, A. bovis, B. canis vogeli and Candidatus Midichloria mitochondrii DNA sequences were detected in R. sanguineus ticks. Candidatus Midichloria mitochondrii DNA was also detected in Hyalomma spp. ticks. Neither Hepatozoon spp. nor Bartonella spp. DNA was detected in any of the ticks examined. This study describes the first detection of E. canis in the tick R. turanicus, which may serve as a vector of this canine pathogen; E. canis was the most common pathogen detected in the collected questing ticks. It also describes the first detection of A. bovis and Candidatus Midichloria mitochondrii in Israel. To the best of the author's knowledge, this is the first report describing the detection of DNA of the latter two pathogens in R. sanguineus, and of A. bovis in R. turanicus.

  15. The Anaplasma marginale msp5 gene encodes a 19-kilodalton protein conserved in all recognized Anaplasma species.

    PubMed Central

    Visser, E S; McGuire, T C; Palmer, G H; Davis, W C; Shkap, V; Pipano, E; Knowles, D P

    1992-01-01

    Immunization with Anaplasma marginale outer membranes induced immunity against clinical disease which correlated with antibody titer to outer membrane proteins, including a 19-kDa protein (N. Tebele, T. C. McGuire, and G. H. Palmer, Infect. Immun. 59:3199-3204, 1991). This 19-kDa protein, designated major surface protein 5 (MSP-5), was encoded by a single-copy 633-bp gene. The molecular mass of MSP-5, defined in immunoblots by binding to monoclonal antibody ANAF16C1, was conserved among all recognized species of Anaplasma: A. marginale, A. centrale, and A. ovis. Recombinant MSP-5, which absorbed the antibody reactivity of bovine immune serum to native MSP-5, was recognized by anti-A. marginale and anti-A. centrale immune sera in a competitive inhibition assay with monoclonal antibody ANAF16C1. The presence of antibody to the epitope defined by monoclonal antibody ANAF16C1 in all postinfection sera tested indicates that this epitope is a potential diagnostic antigen for use in identifying persistently infected cattle. Images PMID:1280624

  16. Molecular detection of Rickettsia, Anaplasma, Coxiella and Francisella bacteria in ticks collected from Artiodactyla in Thailand.

    PubMed

    Sumrandee, Chalao; Baimai, Visut; Trinachartvanit, Wachareeporn; Ahantarig, Arunee

    2016-07-01

    A total of 79 ticks collected from Sambar deer (Cervus unicolor), Barking deer (Muntiacus muntjak) and Wild boar (Sus scrofa) were examined by PCR for the presence of Rickettsia, Anaplasma, Coxiella, and Francisella bacteria. Of the 79 ticks, 13% tested positive for Rickettsia, 15% tested positive for Anaplasma, 4% tested positive for Coxiella, and 3% tested positive for Francisella. Interestingly, triple infection with Anaplasma, Rickettsia and Francisella was determined in a Dermacentor auratus tick. Moreover, another triple infection with Rickettsia, Anaplasma, and Coxiella was found in a Haemaphysalis lagrangei tick. Double infection of Rickettsia with Coxiella was also detected in another H. lagrangei tick. From the phylogenetic analyses, we found a Rickettsia sp. with a close evolutionary relationship to Rickettsia bellii in the H. lagrangei tick. We also found the first evidence of a Rickettsia sp. that is closely related to Rickettsia tamurae in Rhipicephalus (Boophilus) microplus ticks from Thailand. H. lagrangei and Haemaphysalis obesa ticks collected from Sambar deer tested positive for Anaplasma species form the same clade with Anaplasma bovis. In contrast, other H. lagrangei ticks collected from Sambar deer and D. auratus ticks collected from Wild boar were also reported for the first time to be infected with an Anaplasma species that is closely related to Anaplasma platys. The phylogenetic analysis of the 16S rRNA gene of Coxiella bacteria revealed that Coxiella symbionts from H. lagrangei formed a distinctly different lineage from Coxiella burnetii (a human pathogen). Additionally, Francisella bacteria identified in D. auratus ticks were found to be distantly related to a group of pathogenic Francisella species. The identification of these bacteria in several feeding ticks suggests the risk of various emerging tick-borne diseases and endosymbionts in humans, wildlife, and domestic animals in Thailand. Copyright © 2016 Elsevier GmbH. All rights

  17. Improved molecular detection of Ehrlichia and Anaplasma species applied to Amblyomma ticks collected from cattle and sheep in Ethiopia.

    PubMed

    Teshale, S; Geysen, D; Ameni, G; Asfaw, Y; Berkvens, D

    2015-02-01

    Detection of Ehrlichia and Anaplasma species in animals and tick vectors is crucial for an understanding of the epidemiology of diseases caused by these pathogens. In this study, a pair of primers designated EBR2 and EBR3 was designed from the Anaplasma 16S rDNA sequence and was used along with a previously described primer EHR 16SD for the simultaneous detection of Ehrlichia and Anaplasma species by nested PCR. The primers were used to amplify 925bp of DNA from known species of Ehrlichia and Anaplasma. Restriction with MboII and MspI enzymes allowed Ehrlichia and Anaplasma speciation. Restriction with MboII differentiated between An. marginale, Anaplasma (formerly Ehrlichia) sp. Omatjenne, and An. centrale with An. marginale and Anaplasma (formerly Ehrlichia) sp. Omatjenne yielding 2 distinct fragments each while An. centrale produced 3 distinct bands. Ehrlichia ruminantium and An. phagocytophylum remained undigested. Subsequent restriction with MspI differentiated E. ruminantium from An. phagocytophylum with 2 and 4 fragments, respectively. When used on tick samples from the field, 63 ticks (16.4%) out of 384 collected from cattle and sheep were positive for one or more species of Ehrlichia and Anaplasma. The positivity ranged from 6.3% at Andasa to 36.7% at Habernosa. Higher overall infection rates were found in Amblyomma lepidum than in Amblyomma variegatum ticks (p=0.009). Amblyomma lepidum from Habernosa were more often infected with all detected species of Anaplasma and Ehrlichia than Am. variegatum. At Bako, however, Anaplasma (formerly Ehrlichia) sp. Omatjenne was detected only in Am. variegatum. A significantly higher proportion of ticks collected from cattle (20.6%) was found positive than in those collected from sheep (3.3%) (p=0.003). Simultaneous detection of Ehrlichia and Anaplasma species and correct identification of mixed infections was possible. Since the ticks were collected from animals, the occurrence of the major species of Ehrlichia and

  18. "Candidatus anaplasma camelii" in one-humped camels (Camelus dromedarius) in Morocco: a novel and emerging anaplasma species?

    PubMed

    Ait Lbacha, Hicham; Zouagui, Zaid; Alali, Said; Rhalem, Abdelkbir; Petit, Elisabeth; Ducrotoy, Marie Julie; Boulouis, Henri-Jean; Maillard, Renaud

    2017-02-05

    There has been a growing interest in camel anaplasmosis due to its recent emergence in this reservoir species and concerns for its zoonotic potential. The epidemiology of anaplasmosis in camels therefore remains poorly understood mostly because camels belong to marginalised poor and often transhumant populations whose interests are largely neglected. Most studies of anaplasmosis in camels have relied on microscopy and serology for diagnosis and only three studies, undertaken in Tunisia, Saudia Arabia and China, have used molecular diagnostics. The present work characterises Anaplasmataceae strains circulating in the Camelus dromedarius reservoir in Morocco using PCR. Camels (n = 106) were randomly sampled from 6 regions representing different agro-ecological areas in southern Morocco. Whole blood was collected and screened using PCR methods targeting the gene groEL. Anaplasmataceae strains were characterised by sequence analysis of the gene groEL. A total of 39.62% (42/106) camels screened were positive for Anaplasmataceae spp. GenBank BLAST analysis of five positive sequenced samples revealed that all strains were 100% identical to "Candidatus Anaplasma camelii". Phylogenetic investigation and genetic characterisation of the aligned segment (650 bp) of the gene groEL confirmed high similarity with A. platys. This study demonstrates the circulation of a previously unidentified species of the genus Anaplasma in Morocco which is genetically close to the agent causing canine anaplasmosis but whose main reservoir is thought to be Camelus dromedarius. This study is not a clinical trial and therefore a trial registration number does not apply.

  19. Comparative study of Anaplasma parasites in tick carrying buffaloes and cattle

    PubMed Central

    Rajput, Z.I.; Hu, Song-hua; Arijo, A.G.; Habib, M.; Khalid, M.

    2005-01-01

    A comparative study on the prevalence of Anaplasma parasite was conducted on ticks carrying buffaloes and cattle. Five hundred blood samples of both animals (250 of each) were collected during February, March and April. Thin blood smears on glass slides were made, fixed in 100% methyl alcohol and examined. Microscopic examination revealed that 205 (41%) animals had Anaplasma parasites, out of which 89, 44 and 72 animals had Anaplasma marginale, Anaplasma centrale and mixed infection respectively. Infected buffaloes and cattle were 75 and 130 respectively. The infection in female was 53 and 92 in buffaloes and cattle respectively. Twenty-two and 92 blood samples of male were found positive in buffaloes and cattle respectively. Comparative study revealed that the cattle were 26.82% more susceptible than buffaloes. The parasite prevailing percentage in female of both animals was slightly higher than that of the male. This investigation was aimed at studying the comparative prevalence of Anaplasma parasite in tick carrying buffaloes and cattle. PMID:16252338

  20. Molecular survey of Rickettsia, Ehrlichia, and Anaplasma infection of domestic cats in Japan.

    PubMed

    Sasaki, Hiromi; Ichikawa, Yasuaki; Sakata, Yoshimi; Endo, Yasuyuki; Nishigaki, Kazuo; Matsumoto, Kotaro; Inokuma, Hisashi

    2012-12-01

    The prevalence of Rickettsia, Ehrlichia, and Anaplasma in 1764 DNA samples extracted from feline peripheral blood from all 47 prefectures in Japan was evaluated by screening real-time PCR, genus-specific PCR, and DNA nucleotide sequencing. The survey revealed that all cats were negative for Rickettsia infection. Two cats were positive for Ehrlichia or Anaplasma based on the screening PCR assay. Nucleotide sequence analysis of the partial 16S rRNA including the divergent region near the 3'-end revealed that the 2 positives were most similar to Anaplasma bovis with percent identities of 99.8% and 99.2%. This was the first detection of A. bovis DNA fragments in cats. Although these 2 cats showed stomatitis, both were also infected with feline immunodeficiency virus. The relationship between A. bovis carriage and clinical disease is not yet understood.

  1. [Microbiological diagnosis of emerging bacterial pathogens: Anaplasma, Bartonella, Rickettsia, and Tropheryma whipplei].

    PubMed

    Blanco, José Ramón; Jado, Isabel; Marín, Mercedes; Sanfeliu, Isabel; Portillo, Aránzazu; Anda, Pedro; Pons, Immaculada; Oteo, José Antonio

    2008-11-01

    Ehrlichia/Anaplasma, Bartonella, Rickettsia and Tropheryma whipplei (formerly called whippelii) are fastidious bacterial organisms, considered the causative agents of potentially severe emerging and re-emerging diseases with repercussions on public health. The recent availability of advanced molecular biology and cell culture techniques has led to the implication of many of these species in human pathologies. These issues are extensively covered in number 27 of the SEIMC microbiological procedure: Diagnóstico microbiológico de las infecciones por patógenos bacterianos emergentes: Anaplasma, Bartonella, Rickettsia y Tropheryma whippelii (Microbiological diagnosis of Anaplasma, Bartonella, Rickettsia and Tropheryma whippelii infections) (2nd ed., 2007) (www.seimc.org/documentos/protocolos/microbiologia/).

  2. Anaplasma, Ehrlichia and rickettsial pathogens in ixodid ticks infesting cattle and sheep in western Oromia, Ethiopia.

    PubMed

    Teshale, Sori; Kumsa, Bersissa; Menandro, Maria Luisa; Cassini, Rudi; Martini, Marco

    2016-10-01

    Although ticks are widely distributed in all agro-ecological zones of Ethiopia, information on tick-borne pathogens is scarce. This study was conducted to determine the presence of Anaplasma spp., Ehrlichia spp., and Rickettsia spp. in Rhipicephalus evertsi and Rhipicephalus (Boophilus) decoloratus collected from cattle and sheep at Bako, western Oromia, Ethiopia, using polymerase chain reaction and sequencing. Anaplasma ovis and Anaplasma spp., Ehrlichia ruminantium and Ehrlichia spp. were detected in Rh. decoloratus, whereas only A. ovis was detected in Rh. evertsi. Both tick species were found to harbor DNA belonging to Rickettsia spp., and Rickettsia africae. Our findings highlight the risk of infection of animals and humans with these zoonotic tick-borne bacteria in Ethiopia.

  3. Experimental transmission of Anaplasma marginale by male Dermacentor reticulatus

    PubMed Central

    Zivkovic, Zorica; Nijhof, Ard M; de la Fuente, José; Kocan, Katherine M; Jongejan, Frans

    2007-01-01

    Background Bovine anaplasmosis has been reported in several European countries, but the vector competency of tick species for Anaplasma marginale from these localities has not been determined. Because of the wide distributional range of Dermacentor reticulatus within Europe and the major role of Dermacentor spp. as a vector of A. marginale in the United States, we tested the vector competency of D. reticulatus for A. marginale. Results Male D. reticulatus were allowed to feed for 7 days on a calf persistently infected with a Zaria isolate of A. marginale, after which they were removed and held off-host for 7 days. The ticks were then allowed to feed a second time for 7 days on a susceptible tick-naïve calf. Infection of calf No. 4291 was detected 20 days post exposure (p.i.) and confirmed by msp4 PCR. Thirty percent of the dissected acquisition fed ticks was infected. In addition, A. marginale colonies were detected by light microscopy in the salivary glands of the acquisition fed ticks. Transmission of A. marginale to calf No. 9191 was confirmed by examination of Giemsa-stained blood smears and msp4 PCR. Ticks were dissected after transmission feeding and presence of A. marginale was confirmed in 18.5% of the dissected ticks. Conclusion This study demonstrates that D. reticulatus males are competent vectors of A. marginale. Further studies are needed to confirm the vector competency of D. reticulatus for other A. marginale strains from geographic areas in Europe. PMID:18053123

  4. Molecular detection of Ehrlichia canis and Anaplasma platys in dogs in Southern Brazil.

    PubMed

    Lasta, Camila Serina; dos Santos, Andrea Pires; Messick, Joanne Belle; Oliveira, Simone Tostes; Biondo, Alexander Welker; Vieira, Rafael Felipe da Costa; Dalmolin, Magnus Larruscaim; González, Félix Hilario Diaz

    2013-01-01

    The aims of this study were to determine the occurrence of Anaplasma platys and Ehrlichia canis infection in dogs in Porto Alegre, Southern Brazil; and to investigate their association with hematological abnormalities. Serum samples from 196 dogs were first tested using dot-ELISA for antibodies against Anaplasma spp. and Ehrlichia canis. Peripheral blood samples from 199 dogs were subjected to 16S rRNA nested PCR (nPCR) for A. platys and E. canis, followed by DNA sequencing to ensure pathogen identity. A total of 19/196 samples (9.69%) were positive for Anaplasma spp. using ELISA and 28/199 (14.07%) samples were positive for A. platys by nested PCR. All the dog samples were negative for E. canis, both in anti-E. canis antibody tests and in nested PCR. There were no significant differences in hematological parameters between A. platys-PCR positive and negative dogs and Anaplasma spp. serologically positive dogs, except for basophil counts, which were higher in nPCR-positive dogs. This is the first report showing A. platys presence in dogs in Southern Brazil. In conclusion, hematological parameters may not be sufficient to diagnose A. platys infection in dogs in Southern Brazil, probably due either to low pathogenicity or to chronic infection. On the other hand, E. canis may either have very low occurrence or be absent in dogs in Porto Alegre.

  5. Anaplasma marginale actively modulates vacuolar maturation during intracellular infection of its tick vector dermacentor andersoni

    USDA-ARS?s Scientific Manuscript database

    Tick-borne transmission of bacterial pathogens in the Order Rickettsiales is responsible for a diversity of infectious diseases, many of them severe, in both humans and animals. Transmission dynamics differ among these pathogens and are reflected in the pathogen-vector interaction. Anaplasma margina...

  6. The transcription factor relish controls anaplasma marginale infection in the bovine tick rhipicephalus microplus

    USDA-ARS?s Scientific Manuscript database

    Rhipicephalus microplus is an important biological vector of Anaplasma marginale, the etiological agent of bovine anaplasmosis. The knowledge of tick immune responses to control bacterial infections remains limited. In this study, we demonstrate that transcription factor Relish from the Imd signalin...

  7. Protective immunity induced by immunization with a live, cultured anaplasma marginale strain

    USDA-ARS?s Scientific Manuscript database

    Despite significant economic losses resulting from infection with Anaplasma marginale, a tick-transmitted rickettsial disease of cattle, available vaccines provide, at best, only partial protection against clinical disease. The green-fluorescent protein (GFP) expressing mutant of the A. marginale St...

  8. Primary structural variation in anaplasma marginale Msp2 efficiently generates immune escape variants

    USDA-ARS?s Scientific Manuscript database

    Antigenic variation allows microbial pathogens to evade immune clearance and establish persistent infection. Anaplasma marginale utilizes gene conversion of a repertoire of silent msp2 alleles into a single active expression site to encode unique Msp2 variants. As the genomic complement of msp2 alle...

  9. Subdominant outer membrane antigens in anaplasma marginale: conservation, antigenicity, and protective capacity using recombinant protein

    USDA-ARS?s Scientific Manuscript database

    Anaplasma marginale is a tick-borne rickettsial pathogen of cattle with a worldwide distribution. Currently a safe and efficacious vaccine is unavailable. Outer membrane protein (OMP) extracts or a well- defined surface protein complex reproducibly induce protective immunity. However, there are seve...

  10. Anaplasma Infection in Ticks, Livestock and Human in Ghaemshahr, Mazandaran Province, Iran.

    PubMed

    Hosseini-Vasoukolaei, Nasibeh; Oshaghi, Mohammad Ali; Shayan, Parviz; Vatandoost, Hassan; Babamahmoudi, Farhang; Yaghoobi-Ershadi, Mohammad Reza; Telmadarraiy, Zakkyeh; Mohtarami, Fatemeh

    2014-12-01

    Anaplasmosis is an important issue for animal breeders in terms of economic losses as well as a health concern to human. Ticks are considered as the main vector of this disease. Lack of documented information about Anaplasma species in Iran was the scope of this study to determine the population of ticks and the presence of Anaplasma in ticks, domestic ruminants and also human beings in northern Iran. A total of 101 unengorged hard ticks, 78 domestic ruminants and 40 human blood samples collected from Ghaemshahr, Mazandaran Province, northern Iran were tested by nested PCR against 16s rRNA gene of Anaplasma species. Positive PCR was found in 50 ticks, 28 sheep, 2 cattle, one goat, and 10 human specimens. Sequence analysis of the PCR products confirmed presence of A. ovis in two Rhipicephalus sanguineus and two Ixodes ricinus ticks, one human and 4 sheep samples. Moreover one Boophilus annulatus tick and one sheep sample were infected with A. bovis. Furthermore one sample of sheep was infected with A. centrale. This study is the first report of tick infection to A. ovis, A. bovis and human infection to A. ovis in Iran. The result of this study is a survey of Anaplasma infections from ticks, domestic animals and human in Iran which help to have appropriate prevention measures for anaplasmosis.

  11. Association of anaplasma marginale strain superinfection with infection prevalence within tropical regions

    USDA-ARS?s Scientific Manuscript database

    Strain superinfection occurs when a second strain infects a host already infected with a primary strain. The incidence of superinfection with Anaplasma marginale, a tick-borne rickettsial pathogen of domestic and wild ruminants, has been shown to be higher in tropical versus temperate regions. This ...

  12. Knockout of an outer membrane protein operon of anaplasma marginale by transposon mutagenesis

    USDA-ARS?s Scientific Manuscript database

    Large amounts of data generated by genomics, transcriptomics and proteomics technologies have increased our understanding of the biology of Anaplasma marginale. However, these data have also led to new assumptions that require testing, ideally through classic genetic mutation. One example is the def...

  13. Anaplasma Infection in Ticks, Livestock and Human in Ghaemshahr, Mazandaran Province, Iran

    PubMed Central

    Hosseini-Vasoukolaei, Nasibeh; Oshaghi, Mohammad Ali; Shayan, Parviz; Vatandoost, Hassan; Babamahmoudi, Farhang; Yaghoobi-Ershadi, Mohammad Reza; Telmadarraiy, Zakkyeh; Mohtarami, Fatemeh

    2014-01-01

    Background: Anaplasmosis is an important issue for animal breeders in terms of economic losses as well as a health concern to human. Ticks are considered as the main vector of this disease. Lack of documented information about Anaplasma species in Iran was the scope of this study to determine the population of ticks and the presence of Anaplasma in ticks, domestic ruminants and also human beings in northern Iran. Methods: A total of 101 unengorged hard ticks, 78 domestic ruminants and 40 human blood samples collected from Ghaemshahr, Mazandaran Province, northern Iran were tested by nested PCR against 16s rRNA gene of Anaplasma species. Results: Positive PCR was found in 50 ticks, 28 sheep, 2 cattle, one goat, and 10 human specimens. Sequence analysis of the PCR products confirmed presence of A. ovis in two Rhipicephalus sanguineus and two Ixodes ricinus ticks, one human and 4 sheep samples. Moreover one Boophilus annulatus tick and one sheep sample were infected with A. bovis. Furthermore one sample of sheep was infected with A. centrale. Conclusion: This study is the first report of tick infection to A. ovis, A. bovis and human infection to A. ovis in Iran. The result of this study is a survey of Anaplasma infections from ticks, domestic animals and human in Iran which help to have appropriate prevention measures for anaplasmosis. PMID:26114134

  14. Molecular and serological in-herd prevalence of Anaplasma marginale infection in Texas cattle

    USDA-ARS?s Scientific Manuscript database

    Bovine anaplasmosis is an infectious, non-contagious disease caused by the rickettsial pathogen Anaplasma marginale (A. marginale). The organism has a global distribution and infects erythrocytes, resulting in anemia, jaundice, fever, abortions and death. Once infected, animals remain carriers for l...

  15. Multiple infections of Anaplasma platys variants in Philippine dogs

    PubMed Central

    Ybañez, Adrian Patalinghug; Ybañez, Rochelle Haidee Daclan; Yokoyama, Naoaki; Inokuma, Hisashi

    2016-01-01

    Aim: Anaplasma platys, the causative agent of infectious canine cyclic thrombocytopenia, is a tick-borne pathogen that also has been implicated as potentially zoonotic. To provide molecular evidence on the multiple infections of A. platys variants in Philippine dogs. Materials and Methods: DNA fragments of A. platys from infected dogs in the Philippines were molecularly characterized. For screening, 25 dogs suspected to have canine anaplasmosis were tested using a 16S rRNA-based nested polymerase chain reaction (PCR). Infection was confirmed by sequencing of positive amplicons. Second round PCR targeting a longer 16S rRNA fragment was subsequently performed on the first round PCR amplicons of the positive samples. Further characterization using the heat-shock operon (groEL) gene was also performed on the A. platys-positive samples. Results: 10 16S rRNA sequences were obtained and found 99.6-100% identical to each other and 99.6-99.7% identical to the closest registered A. platys sequences. On the other hand, 36 groEL clone sequences were obtained and found to be 85.1-99.8% identical with each other and 85.0-88.9% identical to the closest previously registered A. platys sequences. Four dogs were found coinfected with 2-3 groEL variant sequences. Phylogenetic analyses suggest that the detected A. platys in the Philippines may represent unique variants. Conclusion: A. platys variants were detected in Philippine dogs. Coinfection of different A. platys variants in dogs was also demonstrated. The present study may indicate the potential genetic diversity of A. platys in the country. PMID:28096621

  16. Multiple infections of Anaplasma platys variants in Philippine dogs.

    PubMed

    Ybañez, Adrian Patalinghug; Ybañez, Rochelle Haidee Daclan; Yokoyama, Naoaki; Inokuma, Hisashi

    2016-12-01

    Anaplasma platys, the causative agent of infectious canine cyclic thrombocytopenia, is a tick-borne pathogen that also has been implicated as potentially zoonotic. To provide molecular evidence on the multiple infections of A. platys variants in Philippine dogs. DNA fragments of A. platys from infected dogs in the Philippines were molecularly characterized. For screening, 25 dogs suspected to have canine anaplasmosis were tested using a 16S rRNA-based nested polymerase chain reaction (PCR). Infection was confirmed by sequencing of positive amplicons. Second round PCR targeting a longer 16S rRNA fragment was subsequently performed on the first round PCR amplicons of the positive samples. Further characterization using the heat-shock operon (groEL) gene was also performed on the A. platys-positive samples. 10 16S rRNA sequences were obtained and found 99.6-100% identical to each other and 99.6-99.7% identical to the closest registered A. platys sequences. On the other hand, 36 groEL clone sequences were obtained and found to be 85.1-99.8% identical with each other and 85.0-88.9% identical to the closest previously registered A. platys sequences. Four dogs were found coinfected with 2-3 groEL variant sequences. Phylogenetic analyses suggest that the detected A. platys in the Philippines may represent unique variants. A. platys variants were detected in Philippine dogs. Coinfection of different A. platys variants in dogs was also demonstrated. The present study may indicate the potential genetic diversity of A. platys in the country.

  17. Exploitability Assessment with TEASER

    DTIC Science & Technology

    2017-05-01

    executable xor writable, or that executable images cannot have write permissions over the course of the program’s execution. DWARF Debugging With...feature a Proof of Concept (POC), or input to a program demonstrating the ability to use a bug to exploit the application. For example, a recent write -up...TEASER Exploit Development Workflow Tainted Malloc Reads, Tainted Function Arguments, Tainted Pointer Reads/ Writes , Tainted Data Reads Tainted

  18. A Bayesian spatio-temporal model for forecasting Anaplasma species seroprevalence in domestic dogs within the contiguous United States

    PubMed Central

    Liu, Yan; Watson, Stella C.; Gettings, Jenna R.; Lund, Robert B.; Nordone, Shila K.; McMahan, Christopher S.

    2017-01-01

    This paper forecasts the 2016 canine Anaplasma spp. seroprevalence in the United States from eight climate, geographic and societal factors. The forecast’s construction and an assessment of its performance are described. The forecast is based on a spatial-temporal conditional autoregressive model fitted to over 11 million Anaplasma spp. seroprevalence test results for dogs conducted in the 48 contiguous United States during 2011–2015. The forecast uses county-level data on eight predictive factors, including annual temperature, precipitation, relative humidity, county elevation, forestation coverage, surface water coverage, population density and median household income. Non-static factors are extrapolated into the forthcoming year with various statistical methods. The fitted model and factor extrapolations are used to estimate next year’s regional prevalence. The correlation between the observed and model-estimated county-by-county Anaplasma spp. seroprevalence for the five-year period 2011–2015 is 0.902, demonstrating reasonable model accuracy. The weighted correlation (accounting for different sample sizes) between 2015 observed and forecasted county-by-county Anaplasma spp. seroprevalence is 0.987, exhibiting that the proposed approach can be used to accurately forecast Anaplasma spp. seroprevalence. The forecast presented herein can a priori alert veterinarians to areas expected to see Anaplasma spp. seroprevalence beyond the accepted endemic range. The proposed methods may prove useful for forecasting other diseases. PMID:28738085

  19. The Exploitation of Drug Users.

    PubMed

    Stallings, Shirley; Montagne, Michael

    2015-01-01

    Drug users have been exploited in research studies and clinical practice. We explore ways in which exploitation has occurred and strategies to help patients, research subjects and communities to prevent or avoid exploitation.

  20. Hope and exploitation.

    PubMed

    Martin, Adrienne M

    2008-01-01

    How do we encourage patients to be hopeful without exploiting their hope? A medical researcher or a pharmaceutical company can take unfair advantage of someone's hope by much subtler means than simply giving misinformation. Hope shapes deliberation, and therefore can make deliberation better or worse, by the deliberator's own standards of deliberation.

  1. Anaplasma, Ehrlichia, and "Candidatus Neoehrlichia" bacteria: pathogenicity, biodiversity, and molecular genetic characteristics, a review.

    PubMed

    Rar, Vera; Golovljova, Irina

    2011-12-01

    The family Anaplasmataceae includes gram-negative obligate intracellular alphaproteobacteria, which amplify within membrane-bound vacuoles. The genera Anaplasma and Ehrlichia as well as the recently specified cluster "Candidatus Neoehrlichia" comprise all bacteria of the family that are transmitted by ixodid ticks to mammalian hosts and cause infections in humans and numerous domestic and wild animal species. Totally, six Anaplasma, five Ehrlichia, and two "Candidatus Neoehrlichia" species as well as a variety of Ehrlichia bacteria with undefined taxonomic position belong to tick-borne bacteria. This review briefs the ecology, pathogenic properties, and molecular genetic diversity of Anaplasmataceae validated species, "Candidatus" species, and undefined novel Ehrlichia genetic variants. Copyright © 2011 Elsevier B.V. All rights reserved.

  2. Survey on blood-sucking lice (Phthiraptera: Anoplura) of ruminants and pigs with molecular detection of Anaplasma and Rickettsia spp.

    PubMed

    Hornok, S; Hofmann-Lehmann, R; de Mera, I G Fernández; Meli, M L; Elek, V; Hajtós, I; Répási, A; Gönczi, E; Tánczos, B; Farkas, R; Lutz, H; de la Fuente, J

    2010-12-15

    Lice may serve as biological or mechanical vectors for various infectious agents. To investigate louse infestation of ruminants and pigs, and pathogens potentially transmitted by them, anopluran lice (n=1182) were collected in Hungary, and evaluated for the presence of anaplasma, rickettsia and haemotropic mycoplasma DNA. On cattle the following species were found: Linognathus vituli (57%), Haematopinus eurysternus (38%) and Solenopotes capillatus (5%). L. vituli had a lower mean individual count/host when compared to H. eurysternus. On calves only L. vituli was observed, with a higher louse burden than on full-grown cattle. H. eurysternus and S. capillatus were more likely to occur simultaneously with another species on the same host, than L. vituli. Goats infested with Linognathus stenopsis had the overall highest prevalence (68%), while pigs harbouring Haematopinus suis showed the lowest (<1%). Anaplasma DNA was detected in 50% of pools analysed. In L. vituli Anaplasma ovis (or a closely related novel Anaplasma marginale genotype) was identified. Anaplasma-positivity of H. suis suggests that pigs may extend the reservoir and/or host spectrum of relevant species. Anaplasma-infected L. stenopsis pools show for the first time that caprine anaplasmosis is endemic in Hungary. Rickettsia spp. were demonstrated from Linognathus spp. and H. eurysternus. No haemotropic mycoplasmas were detected in any samples. In conclusion, this is the first molecularly confirmed report of bovine and ovine Anaplasma spp. in L. vituli, L. stenopsis and H. suis. The present results suggest that phthirapterosis of domestic animals deserves more attention, and lice should be evaluated among the broad range of potential vectors of arthropod-borne pathogens.

  3. Molecular characterisation of Anaplasma species from African buffalo (Syncerus caffer) in Kruger National Park, South Africa.

    PubMed

    Sisson, Danielle; Hufschmid, Jasmin; Jolles, Anna; Beechler, Brianna; Jabbar, Abdul

    2017-03-01

    Bovine anaplasmosis is a tick-borne disease, mainly caused by Anaplasma marginale and A. centrale and is distributed in tropical and sub-tropical areas. This study aimed to characterise A. marginale and A. centrale from African buffaloes in Kruger National Park (KNP), South Africa, using the DNA sequences of the genes coding for major surface protein (msp1β) and heat shock protein (groEL), respectively. A total of 747 blood samples were collected from February 2014 to August 2016 from African buffaloes kept in KNP, and DNAs were tested using a molecular-phylogenetic approach. Out of 747 samples tested, 129 (17.3%) and 98 (13.1%) were positive for single infection with A. marginale and A. centrale, respectively; whereas 113 (15.1%) were positive for both Anaplasma spp. Pairwise difference of 1.6-8.5% was observed in msp1β sequences of A. marginale whereas that was only 0.3-2.4% for groEL sequences of A. centrale. Separate phylogenetic analyses of msp1β and groEL sequences of A. marginale and A. centrale, respectively, revealed that sequences of Anaplasma spp. from African buffaloes were unique and they grouped separately when compared with previously published sequences of both species. This is the first study to characterise A. marginale and A. centrale from African buffalo using species specific molecular markers. This study will pave the way for future studies to assess genetic variation among Anaplasma spp. from wild ruminants using molecular markers that are better at differentiating between species and strains than the more commonly used 16S rRNA gene, and help to undertake health and fitness studies and host-parasite dynamics using quantitative molecular tools.

  4. Molecular diagnosis and species identification of Ehrlichia and Anaplasma infections in dogs from Panama, Central America.

    PubMed

    Santamaria, Annamaria; Calzada, Jose E; Saldaña, Azael; Yabsley, Michael J; Gottdenker, Nicole L

    2014-05-01

    The prevalence and distribution of Anaplasma and Ehrlichia were estimated in 201 symptomatic dogs from Panama by nested PCR and DNA sequencing of the 16S rRNA gene. In total, 163 dogs (70.6%; 163/201) were infected with either Anaplasma or Ehrlichia. On the basis of PCR results, the majority of cases were infected with E. canis (64.2%; 129/201) followed by 21.4% (43/201) with A. platys, whereas 7.5% (15/201) had Anaplasma/Ehrlichia co-infections. Further analyses of 16S rDNA partial sequences show sequence homology with E. canis and A. platys from other countries. Hematology findings from 79 E. canis PCR-positive dogs included anemia (74.7%), thrombocytopenia (81.9%), macroplatelets (29.1%), and leukopenia (6.3%). Among 16 A. platys PCR-positive dogs with available hematology, 62.5% were anemic, 75% had thrombocytopenia, and 100% had macroplatelets. On the basis of E.canis serology (enzyme-linked immunosorbent assay [ELISA]) (n=92 dogs), 30 dogs that were seropositive for E. canis were also PCR-positive, whereas among seronegatives (n=62), 10 were PCR-positive for E. canis. This study provides the first characterization of canine anaplasmosis and ehrlichiosis infections in Panama and is important to veterinary public health and comparative studies of these pathogens in the Americas.

  5. Associations between coinfection prevalence of Borrelia lusitaniae, Anaplasma sp., and Rickettsia sp. in hard ticks feeding on reptile hosts.

    PubMed

    Václav, Radovan; Ficová, Martina; Prokop, Pavol; Betáková, Tatiana

    2011-02-01

    An increasing number of studies reveal that ticks and their hosts are infected with multiple pathogens, suggesting that coinfection might be frequent for both vectors and wild reservoir hosts. Whereas the examination of associations between coinfecting pathogen agents in natural host-vector-pathogen systems is a prerequisite for a better understanding of disease maintenance and transmission, the associations between pathogens within vectors or hosts are seldom explicitly examined. We examined the prevalence of pathogen agents and the patterns of associations between them under natural conditions, using a previously unexamined host-vector-pathogen system--green lizards Lacerta viridis, hard ticks Ixodes ricinus, and Borrelia, Anaplasma, and Rickettsia pathogens. We found that immature ticks infesting a temperate lizard species in Central Europe were infected with multiple pathogens. Considering I. ricinus nymphs and larvae, the prevalence of Anaplasma, Borrelia, and Rickettsia was 13.1% and 8.7%, 12.8% and 1.3%, and 4.5% and 2.7%, respectively. The patterns of pathogen prevalence and observed coinfection rates suggest that the risk of tick infection with one pathogen is not independent of other pathogens. Our results indicate that Anaplasma can play a role in suppressing the transmission of Borrelia to tick vectors. Overall, however, positive effects of Borrelia on Anaplasma seem to prevail as judged by higher-than-expected Borrelia-Anaplasma coinfection rates.

  6. Multisensor staring exploitation

    NASA Astrophysics Data System (ADS)

    Bryant, Michael L.

    2008-04-01

    The focus of this paper is on the exploitation of new staring sensors to address the urban surveillance challenge and help combat the war on terror. A staring sensor visualization environment, known as the Data Table, will be presented which integrates staring sensors with close-in sensors, such as small UAVs, building mounted sensors, and unattended ground sensors (UGS). There are several staring sensors in development, but two in particular will be highlighted in this paper - NightStare and the Gotcha Radar, both under development by the Air Force Research Laboratory (AFRL).

  7. Development of duplex PCR for simultaneous detection of Theileria spp. and Anaplasma spp. in sheep and goats.

    PubMed

    Cui, Yanyan; Zhang, Yan; Jian, Fuchun; Zhang, Longxian; Wang, Rongjun; Cao, Shuxuan; Wang, Xiaoxing; Yan, Yaqun; Ning, Changshen

    2017-05-01

    Theileria spp. and Anaplasma spp., which are important tick-borne pathogens (TBPs), impact the health of humans and animals in tropical and subtropical areas. Theileria and Anaplasma co-infections are common in sheep and goats. Following alignment of the relevant DNA sequences, two primer sets were designed to specifically target the Theileria spp. 18S rRNA and Anaplasma spp. 16S rRNA gene sequences. Genomic DNA from the two genera was serially diluted tenfold for testing the sensitivities of detection of the primer sets. The specificities of the primer sets were confirmed when DNA from Anaplasma and Theileria (positive controls), other related hematoparasites (negative controls) and ddH2O were used as templates. Fifty field samples were also used to evaluate the utility of single PCR and duplex PCR assays, and the detection results were compared with those of the PCR methods previously published. An optimized duplex PCR assay was established from the two primer sets based on the relevant genes from the two TBPs, and this assay generated products of 298-bp (Theileria spp.) and 139-bp (Anaplasma spp.). The detection limit of the assay was 29.4 × 10(-3) ng per μl, and there was no cross-reaction with the DNA from other hematoparasites. The results showed that the newly developed duplex PCR assay had an efficiency of detection (P > 0.05) similar to other published PCR methods. In this study, a duplex PCR assay was developed that can simultaneously identify Theileria spp. and Anaplasma spp. in sheep and goats. This duplex PCR is a potentially valuable assay for epidemiological studies of TBPs in that it can detect cases of mixed infections of the pathogens. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. The Geohazards Exploitation Platform

    NASA Astrophysics Data System (ADS)

    Laur, Henri; Casu, Francesco; Bally, Philippe; Caumont, Hervé; Pinto, Salvatore

    2016-04-01

    The Geohazards Exploitation Platform, or Geohazards TEP (GEP), is an ESA originated R&D activity of the EO ground segment to demonstrate the benefit of new technologies for large scale processing of EO data. This encompasses on-demand processing for specific user needs, systematic processing to address common information needs of the geohazards community, and integration of newly developed processors for scientists and other expert users. The platform supports the geohazards community's objectives as defined in the context of the International Forum on Satellite EO and Geohazards organised by ESA and GEO in Santorini in 2012. The GEP is a follow on to the Supersites Exploitation Platform (SSEP) an ESA initiative to support the Geohazards Supersites & Natural Laboratories initiative (GSNL). Today the GEP allows to exploit 70+ Terabyte of ERS and ENVISAT archive and the Copernicus Sentinel-1 data available on line. The platform has already engaged 22 European early adopters in a validation activity initiated in March 2015. Since September, this validation has reached 29 single user projects. Each project is concerned with either integrating an application, running on demand processing or systematically generating a product collection using an application available in the platform. The users primarily include 15 geoscience centres and universities based in Europe: British Geological Survey (UK), University of Leeds (UK), University College London (UK), ETH University of Zurich (CH), INGV (IT), CNR-IREA and CNR-IRPI (IT), University of L'Aquila (IT), NOA (GR), Univ. Blaise Pascal & CNRS (FR), Ecole Normale Supérieure (FR), ISTERRE / University of Grenoble-Alpes (FR). In addition, there are users from Africa and North America with the University of Rabat (MA) and the University of Miami (US). Furthermore two space agencies and four private companies are involved: the German Space Research Centre DLR (DE), the European Space Agency (ESA), Altamira Information (ES

  9. Image exploitation for MISAR

    NASA Astrophysics Data System (ADS)

    Heinze, N.; Edrich, M.; Saur, G.; Krüger, W.

    2007-04-01

    The miniature SAR-system MiSAR has been developed by EADS Germany for lightweight UAVs like the LUNASystem. MiSAR adds to these tactical UAV-systems the all-weather reconnaissance capability, which is missing until now. Unlike other SAR sensors, that produce large strip maps at update rates of several seconds, MiSAR generates sequences of SAR images with approximately 1 Hz frame rate. photo interpreters (PI) of tactical drones, now mainly experienced with visual interpretation, are not used to SARimages, especially not with SAR-image sequence characteristics. So they should be supported to improve their ability to carry out their task with a new, demanding sensor system. We have therefore analyzed and discussed with military PIs in which task MiSAR can be used and how the PIs can be supported by special algorithms. We developed image processing- and exploitation-algorithms for such SAR-image sequences. A main component is the generation of image sequence mosaics to get more oversight. This mosaicing has the advantage that also non straight /linear flight-paths and varying squint angles can be processed. Another component is a screening-component for manmade objects to mark regions of interest in the image sequences. We use a classification based approach, which can be easily adapted to new sensors and scenes. These algorithms are integrated into an image exploitation system to improve the image interpreters ability to get a better oversight, better orientation and helping them to detect relevant objects, especially considering long endurance reconnaissance missions.

  10. Molecular identification and characterization of Anaplasma platys and Ehrlichia canis in dogs in Mexico.

    PubMed

    Almazán, Consuelo; González-Álvarez, Vicente H; Fernández de Mera, Isabel G; Cabezas-Cruz, Alejandro; Rodríguez-Martínez, Rafael; de la Fuente, José

    2016-03-01

    The tick-borne pathogens Ehrlichia canis and Anaplasma platys are the causative agents of canine monocytic ehrlichiosis (CME) and canine cyclic thrombocytopenia (CCT). Although molecular evidence of E. canis has been shown, phylogenetic analysis of this pathogen has not been performed and A. platys has not been identified in Mexico, where the tick vector Rhipicephalus sanguineus sensu lato (s.l.) is common. The aim of this research was to screen, identify and characterize E. canis and A. platys by PCR and phylogenetic analysis in dogs from La Comarca Lagunera, a region formed by three municipalities, Torreon, Gomez-Palacio and Lerdo, in the Northern states of Coahuila and Durango, Mexico. Blood samples and five engorged R. sanguineus s.l. ticks per animal were collected from 43 females and 57 male dogs presented to veterinary clinics or lived in the dog shelter from La Comarca Lagunera. All the sampled dogs were apparently healthy and PCR for Anaplasma 16S rRNA, Ehrlichia 16S rRNA, and E. canis trp36 were performed. PCR products were sequenced and used for phylogenetic analysis. PCR products were successfully amplified in 31% of the samples using primers for Anaplasma 16S rRNA, while 10% and 4% amplified products using primers for Ehrlichia 16S rRNA and E. canis trp36 respectively. Subsequent sequencing and phylogenetic analyses of these products showed that three samples corresponded to A. platys and four to E. canis. Based on the analysis of trp36 we confirmed that the E. canis strains isolated from Mexico belong to a conservative clade of E. canis and are closely related to strains from USA. In conclusion, this is the first molecular identification of A. platys and the first molecular characterization and phylogenetic study of both A. platys and E. canis in dogs in Mexico.

  11. Novel Anaplasma and Ehrlichia organisms infecting the wildlife of two regions of the Brazilian Amazon.

    PubMed

    Soares, Herbert S; Marcili, Arlei; Barbieri, Amália R M; Minervino, Antonio H H; Malheiros, Antonio F; Gennari, Solange M; Labruna, Marcelo B

    2017-10-01

    During 2009-2012, wild animals were sampled in the Amazon biome of Brazil. Animal tissues and blood were tested by polymerase chain reaction (PCR) assays targeting DNA of the bacterial family Anaplasmataceae (genera Anaplasma, Ehrlichia, Wolbachia) and the genus Borrelia. Overall, 181 wild animals comprising 36 different species (2 reptiles, 5 birds, and 29 mammals) were sampled. All birds and reptiles were negative by all PCR assays, as well as all mammals for the Borrelia PCR assay. Anaplasmataceae agents were searched by PCR assays targeting two different genes, the ribosomal 16S rRNA gene and the protein-coding dsb gene. Three dsb closely related haplotypes were generated from 3 white-lipped peccaries (Tayassu pecari). In a phylogenetic analysis inferred from dsb partial sequences, these haplotypes grouped with previously reported Ehrlichia haplotypes from jaguar (Panthera onca) and horse from Brazil, suggesting that they could all represent a single species, yet to be properly characterized. A unique dsb haplotype was generated from a sloth (Bradypus tridactylus), and could also represent a different Ehrlichia species. All these dsb haplotypes formed a clade sister to the Ehrlichia ruminantium clade. Three distinct 16S rRNA gene haplotypes were generated from a wild guinea pig (Cavia sp.), a woolly mouse opossum (Micoureus demerarae), and two from robust capuchin monkeys (Sapajus sp.). In a phylogenetic analysis inferred from 16S rRNA gene partial sequence, these haplotypes grouped within the Wolbachia clade, and are likely to represent Wolbachia organisms that were infecting invertebrate metazoarians (e.g., filarids) associated with the sampled mammals. Two deer (Mazama americana) samples yielded two distinct 16S rRNA gene sequences, one identical to several sequences of Anaplasma bovis, and an unique sequence that grouped in a clade with different Anaplasma species. Our results indicate that a variety of genetically distinct Anaplasmataceae organisms

  12. Survey of Ticks Collected from Tennessee Cattle and Their Pastures for Anaplasma and Ehrlichia Species.

    PubMed

    Pompo, K; Mays, S; Wesselman, C; Paulsen, D J; Fryxell, R T Trout

    2016-02-01

    Anaplasma marginale is the causative agent for bovine anaplasmosis (BA) and Ehrlichia ruminantium is the causative agent for heartwater, 2 devastating diseases of cattle. BA is common in the United States and frequently reported in western Tennessee cattle; however, cases of heartwater are not yet established in the continental United States. Because both pathogens are transmitted via the bites of infected ticks, the objective of this study was to survey cattle and pastures for ticks and for each pathogen. University of Tennessee AgResearch has 7 research and education centers (REC) located throughout the state at which they manage cattle. Ticks were collected from selected cattle (every fourth to sixth animal) and pastures (via dragging) associated with the herd from each REC during the summer of 2013. A total of 512 ticks were collected from cattle (n = 386) and pastures (n = 126) and were PCR-screened for Anaplasma and Ehrlichia using genus-specific primers. Collections consisted of 398 (77.7%) Amblyomma americanum, 84 (16.4%) Amblyomma maculatum, and 30 (5.9%) Dermacentor variabilis. Ticks were not recovered from pastures or cattle east of the Tennessee Plateau. The North American vectors for An. marginale and E. ruminantium were identified (D. variabilis and A. maculatum, respectively), but neither pathogen was recovered. A large proportion of ticks were collected from cattle and, of these, a majority were attached to their host (compared to questing on their host or engorged on the host). Four A. americanum were positive for Ehrlichia spp. (Ehrlichia ewingii, Ehrlichia chaffeensis, and Panola Mountain Ehrlichia), all in western Tennessee. With the identification of a few Ehrlichia infections in cattle-associated ticks and current A. marginale rates in Tennessee beef cattle nearing 11%, additional research is needed to establish baseline tick, Anaplasma, and Ehrlichia data for future management studies.

  13. Validation of an Anaplasma marginale cELISA for use in the diagnosis of A. ovis infections in domestic sheep and Anaplasma spp. in wild ungulates.

    PubMed

    Scoles, Glen A; Goff, Will L; Lysyk, Timothy J; Lewis, Gregory S; Knowles, Donald P

    2008-07-27

    A commercially available (cELISA) kit for diagnosing Anaplasma marginale infection in cattle was validated for diagnosing A ovis infection in sheep using the bovine serum controls as supplied by the manufacturer (BcELISA) and sheep serum controls from pathogen-free sheep (OcELISA). True positives were identified using two previously established assays, a nested PCR (nPCR) test and an indirect immunofluorescent assay (IFA). The BcELISA was also applied to sera from various species of wild ruminants, comparing the results with the IFA. Receiver operating characteristic (ROC) analysis indicated that the predicted threshold inhibition for the BcELISA was 19.2. The sensitivity for the BcELISA was 98.2% and the specificity was 96.3%. The predicted threshold inhibition decreased to 14.3 for the OcELISA; the sensitivity was 96.5% and the specificity was 98.1%. There was >/=90% concordance between IFA and nPCR, as well as between the BcELISA at 19% inhibition cutoff and either IFA or PCR. Concordance between the cELISA and IFA using sera from elk, mule deer, bighorn sheep, pronghorn antelope, and black-tailed deer ranged from 64% to 100%. This commercially available cELISA test kit can be used very effectively to test domestic sheep for infection with A. ovis using the kit-supplied controls (i.e. the BcELISA) and a 19% inhibition cutoff; the kit may also be useful for detecting intra-erythrocytic Anaplasma infections in wild ruminants.

  14. Cell tropism and molecular epidemiology of Anaplasma platys-like strains in cats.

    PubMed

    Zobba, R; Anfossi, A G; Visco, S; Sotgiu, F; Dedola, C; Pinna Parpaglia, M L; Battilani, M; Pittau, M; Alberti, A

    2015-04-01

    Bacterial species of the genus Anaplasma are tick transmitted pathogens that negatively impact on animal productions and generate veterinary and public health concerns. This paper reports the identification, molecular characterization and phylogeny of novel unclassified A. platys-like strains in cats. Interestingly, these novel strains are closely related to conspecific strains recently identified in ruminants, and significantly differ from A. platys. A. platys-like strains in cats, unlike ruminants strains, show tropism for platelets. Results have implications in the diagnostic scenario of animal anaplasmosis and provide background for reconstructing the evolutionary history of species genetically related to A. platys. Copyright © 2015 Elsevier GmbH. All rights reserved.

  15. Case report: Molecular evidence of Anaplasma platys infection in two women from Venezuela.

    PubMed

    Arraga-Alvarado, Cruz M; Qurollo, Barbara A; Parra, Omaira C; Berrueta, Maribel A; Hegarty, Barbara C; Breitschwerdt, Edward B

    2014-12-01

    This article presents two case reports of Anaplasma platys detection in two women from Venezuela. Both patients were exposed to Rhipicephalus sanguineus, the presumed tick vector, and experienced chronic, nonspecific clinical signs including headaches and muscle pains. Intra-platelet inclusion bodies resembling A. platys were observed in buffy coat smears and A. platys DNA was amplified and sequenced from whole blood; however, treatment with doxycycline did not alleviate their symptoms. These cases provide further support for A. platys as a zoonotic tick-borne pathogen, most likely of low pathogenicity; nonetheless, the cause of illness in humans by A. platys is yet to be confirmed.

  16. Molecular Evidence of Anaplasma platys Infection in Two Women from Venezuela

    PubMed Central

    Arraga-Alvarado, Cruz M.; Qurollo, Barbara A.; Parra, Omaira C.; Berrueta, Maribel A.; Hegarty, Barbara C.; Breitschwerdt, Edward B.

    2014-01-01

    This article presents two case reports of Anaplasma platys detection in two women from Venezuela. Both patients were exposed to Rhipicephalus sanguineus, the presumed tick vector, and experienced chronic, nonspecific clinical signs including headaches and muscle pains. Intra-platelet inclusion bodies resembling A. platys were observed in buffy coat smears and A. platys DNA was amplified and sequenced from whole blood; however, treatment with doxycycline did not alleviate their symptoms. These cases provide further support for A. platys as a zoonotic tick-borne pathogen, most likely of low pathogenicity; nonetheless, the cause of illness in humans by A. platys is yet to be confirmed. PMID:25266347

  17. Anaplasma marginale Yucatan (Mexico) Strain. Assessment of low virulence and potential use as a live vaccine.

    PubMed

    Rodríguez Camarillo, Sergio D; García Ortiz, Miguel Angel; Rojas Ramírez, Edmundo E; Cantó Alarcón, Germinal J; Preciado de la Torre, Jesús F; Rosario Cruz, Rodrigo; Ramos Aragón, Juan A; Aboytes Torres, Ramón

    2008-12-01

    Anaplasma marginale Yucatan strain was found to have low virulence in cattle. We studied the virulence of this isolate by experimental inoculation of 113 susceptible cattle at increasing doses, after which only one animal required treatment for clinical disease. Subsequently, 104 cattle received a live vaccine of this strain by inoculation, which induced immunoprotection after heterologous challenged exposure with a different A. marginale isolate. In this study 14% of the immunized cattle required treatment as compared with the control nonimmunized cattle, in which 56% required treatment. The A. marginale vaccine strains used for the immunization studies had MSP1a variable regions that were different from those used for the challenge exposure.

  18. Cooperation of Pd-1 and LAG-3 contributes to T-cell exhaustion in anaplasma marginale-infected cattle

    USDA-ARS?s Scientific Manuscript database

    The CD4+ T-cell response is central for control of Anaplasma marginale infection in cattle. However, the infection induces a functional exhaustion of antigen-specific CD4+ T cells in cattle immunized with A. marginale outer membrane proteins or purified outer membranes (OM), which presumably facilit...

  19. Identification of multi-locus genetic heterogeneity in anaplasma marginale ss. centrale and its restriction following tick-borne transmission.

    USDA-ARS?s Scientific Manuscript database

    Anaplasma marginale ss. centrale was the first vaccine used to protect against a rickettsial disease and continues in widespread use a century after initial implementation. As its use preceded development of either cryopreservation or cell culture, the vaccine strain was maintained for decades by se...

  20. Genetic profiling for Anaplasma and Ehrlichia species in ticks collected in the Eastern Cape Province of South Africa.

    PubMed

    Iweriebor, Benson C; Mmbaga, Elia J; Adegborioye, Abiodun; Igwaran, Aboi; Obi, Larry C; Okoh, Anthony I

    2017-02-27

    Anaplasma and Ehrlichia are emerging tick-borne pathogens that cause anaplasmosis and ehrlichiosis in humans and other animals worldwide. Infections caused by these pathogens are deadly if left untreated. There has been relatively no systematic survey of these pathogens among ticks in South Africa, thus necessitating this study. The presence of Anaplasma and Ehrlichia species were demonstrated by PCR in ticks collected from domestic ruminants at some selected communities in the Eastern Cape of South Africa. The ticks were identified by morphological characteristics and thereafter processed to extract bacterial DNA, which was analyzed for the presence of genetic materials of Anaplasma and Ehrlichia. Three genera of ticks comprising five species were identified. The screening yielded 16 positive genetic materials that were phylogenetically related to Ehrlichia sequences obtained from GenBank, while no positive result was obtained for Anaplasma. The obtained Ehrlichia sequences were closely related to E. chaffeensis, E. canis, E. muris and the incompletely described Ehrlichia sp. UFMG-EV and Ehrlichia sp. UFMT. The findings showed that ticks in the studied areas were infected with Ehrlichia spp. and that the possibility of transmission to humans who might be tick infested is high.

  1. Tick-Borne Transmission of Two Genetically Distinct Anaplasma marginale Strains following Superinfection of the Mammalian Reservoir Host

    USDA-ARS?s Scientific Manuscript database

    Strain superinfection affects the dynamics of epidemiological spread of pathogens through a host population. Superinfection has recently been shown to occur for genetically distinct strains of the tick-borne pathogen Anaplasma marginale that encode distinctly different surface protein variants. Supe...

  2. Expression of Anaplasma marginale ankyrin repeat-containing proteins during infection of the mammalian host and tick vector

    USDA-ARS?s Scientific Manuscript database

    Using searches of the NCBI conserved domain database and SMART genomic architecture analysis, we identified three ankyrin repeat-containing genes in Anaplasma marginale: AM705, AM926 and AM638. Recombinant protein was used to immunize mice and generate fusion hybridomas secreting protein-specific mo...

  3. First description of natural Ehrlichia canis and Anaplasma platys infections in dogs from Argentina.

    PubMed

    Eiras, Diego Fernando; Craviotto, María Belén; Vezzani, Darío; Eyal, Osnat; Baneth, Gad

    2013-03-01

    Bacteria belonging to the Anaplasmataceae family are vector transmitted agents that affect a variety of vertebrate hosts including the tick-borne pathogens Ehrlichia canis and Anaplasma platys, which cause canine monocytic ehrlichiosis and cyclic thrombocytopenia, respectively. These two infections, typically reported from tropical and sub-tropical regions, have not been previously reported in dogs from Argentina. A total of 86 blood samples from dogs with suspected rickettsial disease and 28 non-suspected dogs were studied. Analysis included evaluation of hematological findings, PCR for Ehrlichia and Anaplasma species and sequencing of the positive PCR products. E. canis was detected in the blood of six dogs and A. platys in eighteen. All the dogs categorized as non-suspected were negative by PCR. Co-infection with Hepatozoon canis and Babesia vogeli was documented. This first report of E. canis and A. platys infections in dogs from Argentina indicates that these tick-borne infections have a considerably broader range than previously recognized in South America.

  4. Molecular and serological prevalence of Anaplasma marginale in water buffaloes in northern Brazil.

    PubMed

    Barbosa da Silva, Jenevaldo; Vinhote, Wagner Marcelo Sousa; Oliveira, Carlos Magno Chaves; André, Marcos Rogério; Machado, Rosangela Zacarias; da Fonseca, Adivaldo Henrique; Barbosa, José Diomedes

    2014-03-01

    Bovine anaplasmosis, caused by Anaplasma marginale, occurs in tropical and subtropical regions throughout the world and is a major constraint on cattle production in many countries. Approximately 60% of the buffalo herds in South America are located in northern Brazil. However, compared with the research on cattle, research on buffaloes has been neglected. Therefore, the present study was conducted to investigate the distribution of A. marginale in water buffaloes in northern Brazil. A total of 500 buffalo blood samples was randomly collected from 16 provinces and was analyzed using both nPCR assay and ELISA techniques. The percentage of animals that were seropositive for A. marginale according to ELISA was 49% (245/500). The main risk factors associated with seroprevalence were the region (p=0.021; OR=1.2) and the reproductive status (p=0.0001; OR=1.6). Anaplasma marginale DNA was detected in 5.4% (27/500) of the sampled buffaloes. Our data provide information about the incidence of A. marginale infection in water buffaloes and may guide future programs aimed at controlling the disease in the northern region of Brazil. Although these water buffaloes are exposed to A. marginale, a low rate of A. marginale PCR-positive animals was found, which could be explained by the habitat in which the sampled animals live because they exhibited a low rate of attached ticks on their skin. Copyright © 2013 Elsevier GmbH. All rights reserved.

  5. Coinfection of sheep with Anaplasma, Theileria and Babesia species in the Kurdistan Region, Iraq.

    PubMed

    Renneker, S; Abdo, J; Bakheit, M A; Kullmann, B; Beyer, D; Ahmed, J; Seitzer, U

    2013-11-01

    Infections of small ruminants with Anaplasma, Theileria and Babesia species are widely distributed in the old world and are of great economic impact. In Iraq, data on disease occurrence in sheep caused by above-mentioned infectious agents are scarce. This study provides information on various haemoparasitic agents infecting sheep in the Kurdistan Region, Iraq, using molecular diagnostic tools. Altogether, 195 samples originating from three governorates in the Kurdistan Region, namely Duhok, Erbil and Sulaimaniya, were analysed. The following pathogens were identified: Anaplasma ovis (62.6%), Theileria ovis (14.35%), T. lestoquardi (7.7%), T. uilenbergi (5.6%) and Babesia ovis (1.5%). T. uilenbergi is detected for the first time in Iraq. Coinfection of sheep with different pathogens could be observed in this study, and it was found that 45 of 195 (23%) of the samples contained more than one pathogen. Even triple-positive samples were identified in 3% of the investigated animals. In conclusion, we confirm the coinfection of sheep with various haemoparasitic pathogen species in the Kurdistan Region of Iraq. Further investigations are needed to reveal the epidemiology of the diseases, the respective tick vectors, and, in the case of coinfection, pathogens' interaction and possible cross-protection.

  6. AMU NEXRAD Exploitation Task

    NASA Technical Reports Server (NTRS)

    Lambert, Winifred C.; Wheeler, Mark M.

    1997-01-01

    This report documents the results of the Applied Meteorology Unit's NEXRAD Exploitation Task. The objectives of this task are to determine what radar signatures are present prior to and at the time of convection initiation, and to determine radar signatures which will help distinguish whether the ensuing convection will become severe. Radar data from the WSR-88D radar located at NWS Melbourne (WSR-88D/KMLB) were collected between June and September 1995, and 16 convective case studies were analyzed for which the radar was operating during the entire period of interest. All WSR-88D/KMLB products were scrutinized for their utility in detecting convection initiation and severe storm signatures. Through process of elimination, it was found that the 0.5 deg reflectivity product with the lowest reflectivity values displayed is the best product to monitor for convection initiation signatures. Seven meteorological features associated with the initiation of deep convection were identified: the Merritt Island and Indian River convergence zones, interlake convergence, horizontal convective rolls, the sea breeze, storm outflow boundaries, and fires. Their reflectivity values ranged from -5 to 20 dBZ. Of the three severe weather phenomena (winds greater than or equal to 50 kts, tornado, 3/4 inch hail), high wind events due to microbursts were most common in the data set. It was found that the values and trends of composite reflectivity, vertically integrated liquid, and core aspect ratio were key indicators of the potential of a cell to produce a microburst. The data were not analyzed for the other two severe weather phenomena because they rarely occurred during the data collection period. This report also includes suggestions for new WSR-88D products, summaries of ongoing research aimed at creating new products, and explicit recommended procedures for detecting convection initiation and severe storm signatures in the radar data using the currently available technology.

  7. Expression of Heat Shock and Other Stress Response Proteins in Ticks and Cultured Tick Cells in Response to Anaplasma spp. Infection and Heat Shock

    PubMed Central

    Villar, Margarita; Ayllón, Nieves; Busby, Ann T.; Galindo, Ruth C.; Blouin, Edmour F.; Kocan, Katherine M.; Bonzón-Kulichenko, Elena; Zivkovic, Zorica; Almazán, Consuelo; Torina, Alessandra; Vázquez, Jesús; de la Fuente, José

    2010-01-01

    Ticks are ectoparasites of animals and humans that serve as vectors of Anaplasma and other pathogens that affect humans and animals worldwide. Ticks and the pathogens that they transmit have coevolved molecular interactions involving genetic traits of both the tick and the pathogen that mediate their development and survival. In this paper, the expression of heat shock proteins (HSPs) and other stress response proteins (SRPs) was characterized in ticks and cultured tick cells by proteomics and transcriptomics analyses in response to Anaplasma spp. infection and heat shock. The results of these studies demonstrated that the stress response was activated in ticks and cultured tick cells after Anaplasma spp. infection and heat shock. However, in the natural vector-pathogen relationship, HSPs and other SRPs were not strongly activated, which likely resulted from tick-pathogen coevolution. These results also demonstrated pathogen- and tick-specific differences in the expression of HSPs and other SRPs in ticks and cultured tick cells infected with Anaplasma spp. and suggested the existence of post-transcriptional mechanisms induced by Anaplasma spp. to control tick response to infection. These results illustrated the complexity of the stress response in ticks and suggested a function for the HSPs and other SRPs during Anaplasma spp. infection. PMID:22084679

  8. Babesia canis vogeli, Ehrlichia canis, and Anaplasma platys infection in a dog.

    PubMed

    Al Izzi, Salah; Martin, Donald S; Chan, Roxanne Y Y; Leutenegger, Christian M

    2013-12-01

    A 12-month-old male neutered mixed breed dog was presented with a history of diarrhea, lethargy, emaciation, polydypsia, and sniffling. Physical examination findings included pale mucous membranes, increased heart and respiratory rates, and normal rectal temperature (38°C). Hematologic abnormalities included anemia and thrombocytopenia. Biochemical abnormalities included hypoalbuminemia, hyperbilirubinemia, and elevated ALP and ALT activities. A SNAP 4Dx test result was positive for Ehrlichia canis. Babesia canis vogeli organisms were found in the peripheral blood films, while morulae of E canis were not seen. Real-time polymerase chain reaction testing confirmed the presence of both B c vogeli and E canis organisms, and also was positive for Anaplasma platys infection. The dog recovered following treatment with doxycycline and imidocarb dipropionate, with normal hematology and biochemical profiles.

  9. First molecular detection of Ehrlichia canis and Anaplasma platys in ticks from dogs in Cebu, Philippines.

    PubMed

    Ybañez, Adrian P; Perez, Zandro O; Gabotero, Shirleny R; Yandug, Ryan T; Kotaro, Matsumoto; Inokuma, Hisashi

    2012-12-01

    Ehrlichia canis infection of dogs in the Philippines has been detected by serological and peripheral blood smear examination methods, but not by molecular means. Anaplasma platys infection in dogs has not yet been officially reported, although it is suspected to occur in the country. Thus, sensitive and specific molecular techniques were used in this study to demonstrate the presence of both E. canis and A. platys in the Philippines. A total of 164 Rhipicephalus sanguineus ticks was collected from 36 dogs. Seven tick samples were found positive with E. canis and one sample with A. platys. To further characterize these pathogens, molecular analyses based on citrate synthase and heat-shock operon genes were also performed. Philippine strains were found to be not divergent from strains from other countries. The present results are the first molecular detection and analyses of E. canis and A. platys in ticks from dogs in the Philippines.

  10. Rickettsia, Ehrlichia, Anaplasma, and Bartonella in ticks and fleas from dogs and cats in Bangkok.

    PubMed

    Foongladda, Suporn; Inthawong, Dutsadee; Kositanont, Uraiwan; Gaywee, Jariyanart

    2011-10-01

    Flea and tick specimens (5-10 fleas or ticks) on dogs and cats from various sites in Bangkok were tested by polymerase chain reaction and DNA sequencing to detect DNA of bacteria Rickettsia (gltA and 17 kDa genes), Anaplasmataceae (16S rRNA gene), and Bartonella (pap31 and its genes). We confirmed that Rickettsia sp. related to Rickettsia felis was detected in 66 of 98 (67.4%) flea specimens from dogs, whereas 8 Bartonella henselae and 2 Bartonella clarridgeiae were detected in 10 of 54 (18.5%) flea specimens from cats. Further, this work provides the first evidence of 10 Ehrlichia canis (3.3%), 7 Anaplasma platys (2.3%), and 2 Wolbachia spp. (0.66%) in 304 Rhipicephalus sanguineus tick specimens in Thailand.

  11. Seasonal Dynamics of Anaplasma phagocytophila in a Rodent-Tick (Ixodes trianguliceps) System, United Kingdom

    PubMed Central

    Begon, Michael; Bennett, Malcolm; Woldehiwet, Zerai; Ogden, Nicholas H.

    2003-01-01

    We investigated the reservoir role of European wild rodents for Anaplasma phagocytophila using polymerase chain reaction (PCR) analysis of blood collected from individually tagged rodents captured monthly over 2 years. The only tick species observed in the woodland study site was Ixodes trianguliceps, and ruminant reservoir hosts were not known to occur. A. phagocytophila infections were detected in both bank voles and wood mice but were restricted to periods of peak nymphal and adult tick activity. Most PCR-positive rodents were positive only once, suggesting that rodent infections are generally short-lived and that ticks rather than rodents may maintain the infection over winter. Bank voles were more likely to be PCR positive than wood mice, possibly because detectable infections are longer lived in bank voles. This study confirms that woodland rodents can maintain A. phagocytophila in Great Britain in the absence of other reservoir hosts and suggests that I. trianguliceps is a competent vector. PMID:12533283

  12. Epidemiology of Babesia, Anaplasma and Trypanosoma species using a new expanded reverse line blot hybridization assay.

    PubMed

    Paoletta, Martina Soledad; López Arias, Ludmila; de la Fournière, Sofía; Guillemi, Eliana Carolina; Luciani, Carlos; Sarmiento, Néstor Fabián; Mosqueda, Juan; Farber, Marisa Diana; Wilkowsky, Silvina Elizabeth

    2017-09-01

    Vector-borne hemoparasitic infections are a major problem that affects livestock industries worldwide, particularly in tropical and subtropical regions. In this work, a reverse line blot (RLB) hybridization assay was developed for the simultaneous detection and identification of Anaplasma, Babesia and bovine trypanosomes, encompassing in this way the most relevant hemoparasites that affect cattle. A total of 186 bovine blood samples collected from two different ecoepidemiological regions of northeast Argentina, with and without tick control, were analyzed with this new RLB. High diversity of parasites, such as Babesia bovis, B. bigemina, Anaplasma marginale and three different Trypanosoma species, was found. High rates of coinfections were also detected, and significant differences were observed not only in the prevalence of parasites but also in the level of coinfections between the two analyzed areas. Regarding the Trypanosoma genus, we provide molecular evidence of the presence of T. vivax and T. theileri for the first time in Argentina. Besides, since the RLB is a prospective tool, it allowed the identification of a yet unknown bovine trypanosome which could not be assigned to any of the bovine species known so far. In the present study we provide new insights on the prevalence of several pathogens that directly impact on livestock production in Argentina. The RLB assay developed here allows to identify simultaneously numerous pathogenic species which can also be easily expanded to detect other blood borne pathogens. These characteristics make the RLB hybridization assay an essential tool for epidemiological survey of all vector-borne pathogens. Copyright © 2017 Elsevier GmbH. All rights reserved.

  13. Propagation of the Israeli vaccine strain of Anaplasma centrale in tick cell lines.

    PubMed

    Bell-Sakyi, Lesley; Palomar, Ana M; Bradford, Emma L; Shkap, Varda

    2015-09-30

    Anaplasma centrale has been used in cattle as a live blood vaccine against the more pathogenic Anaplasma marginale for over 100 years. While A. marginale can be propagated in vitro in tick cell lines, facilitating studies on antigen production, immunisation and vector-pathogen interaction, to date there has been no in vitro culture system for A. centrale. In the present study, 25 cell lines derived from 13 ixodid tick species were inoculated with the Israeli vaccine strain of A. centrale and monitored for at least 12 weeks by microscopic examination of Giemsa-stained cytocentrifuge smears. Infection of 19 tick cell lines was subsequently attempted by transfer of cell-free supernate from vaccine-inoculated tick cells. In two separate experiments, rickettsial inclusions were detected in cultures of the Rhipicephalus appendiculatus cell line RAE25 28-32 days following inoculation with the vaccine. Presence of A. centrale in the RAE25 cells was confirmed by PCR assays targeting the 16S rRNA, groEL and msp4 genes; sequenced PCR products were 100% identical to published sequences of the respective genes in the Israeli vaccine strain of A. centrale. A. centrale was taken through three subcultures in RAE25 cells over a 30 week period. In a single experiment, the Dermacentor variabilis cell line DVE1 was also detectably infected with A. centrale 11 weeks after inoculation with the vaccine. Availability of an in vitro culture system for A. centrale in tick cells opens up the possibility of generating a safer and more ethical vaccine for bovine anaplasmosis. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  14. Bacterial Profiling Reveals Novel "Ca. Neoehrlichia", Ehrlichia, and Anaplasma Species in Australian Human-Biting Ticks.

    PubMed

    Gofton, Alexander W; Doggett, Stephen; Ratchford, Andrew; Oskam, Charlotte L; Paparini, Andrea; Ryan, Una; Irwin, Peter

    2015-01-01

    In Australia, a conclusive aetiology of Lyme disease-like illness in human patients remains elusive, despite growing numbers of people presenting with symptoms attributed to tick bites. In the present study, we surveyed the microbial communities harboured by human-biting ticks from across Australia to identify bacteria that may contribute to this syndrome. Universal PCR primers were used to amplify the V1-2 hyper-variable region of bacterial 16S rRNA genes in DNA samples from individual Ixodes holocyclus (n = 279), Amblyomma triguttatum (n = 167), Haemaphysalis bancrofti (n = 7), and H. longicornis (n = 7) ticks. The 16S amplicons were sequenced on the Illumina MiSeq platform and analysed in USEARCH, QIIME, and BLAST to assign genus and species-level taxonomies. Nested PCR and Sanger sequencing were used to confirm the NGS data and further analyse novel findings. All 460 ticks were negative for Borrelia spp. by both NGS and nested PCR analysis. Two novel "Candidatus Neoehrlichia" spp. were identified in 12.9% of I. holocyclus ticks. A novel Anaplasma sp. was identified in 1.8% of A. triguttatum ticks, and a novel Ehrlichia sp. was identified in both A. triguttatum (1.2%) ticks and a single I. holocyclus (0.6%) tick. Further phylogenetic analysis of novel "Ca. Neoehrlichia", Anaplasma and Ehrlichia based on 1,265 bp 16S rRNA gene sequences suggests that these are new species. Determining whether these newly discovered organisms cause disease in humans and animals, like closely related bacteria do abroad, is of public health importance and requires further investigation.

  15. Co-infection with Anaplasma platys, Bartonella henselae and Candidatus Mycoplasma haematoparvum in a veterinarian.

    PubMed

    Maggi, Ricardo G; Mascarelli, Patricia E; Havenga, Lauren N; Naidoo, Vinny; Breitschwerdt, Edward B

    2013-04-15

    During a two year period, a 27-year-old female veterinarian experienced migraine headaches, seizures, including status epilepticus, and other neurological and neurocognitive abnormalities. Prior to and during her illness, she had been actively involved in hospital-based work treating domestic animals, primarily cats and dogs, in Grenada and Ireland and anatomical research requiring the dissection of wild animals (including lions, giraffe, rabbits, mongoose, and other animals), mostly in South Africa. The woman reported contact with fleas, ticks, lice, biting flies, mosquitoes, spiders and mites and had also been scratched or bitten by dogs, cats, birds, horses, reptiles, rabbits and rodents. Prior diagnostic testing resulted in findings that were inconclusive or within normal reference ranges and no etiological diagnosis had been obtained to explain the patient's symptoms. PCR assays targeting Anaplasma sp. Bartonella sp. and hemotopic Mycoplasma sp. were used to test patient blood samples. PCR positive amplicons were sequenced directly and compared to Gen Bank sequences. In addition, Bartonella alpha Proteobacteria growth medium (BAPGM) enrichment blood culture was used to facilitate bacterial growth and Bartonella spp. serology was performed by indirect fluorescent antibody testing. Anaplasma platys, Bartonella henselae and Candidatus Mycoplasma haematoparvum DNA was amplified and sequenced from the woman's blood, serum or blood culture samples. Her serum was variably seroreactive to several Bartonella sp. antigens. Despite symptomatic improvement, six months of doxycycline most likely failed to eliminate the B. henselae infection, whereas A. platys and Candidatus M. haematoparvum DNA was no longer amplified from post-treatment samples. As is typical of many veterinary professionals, this individual had frequent exposure to arthropod vectors and near daily contact with persistently bacteremic reservoir hosts, including cats, the primary reservoir host for B

  16. Co-infection with Anaplasma platys, Bartonella henselae and Candidatus Mycoplasma haematoparvum in a veterinarian

    PubMed Central

    2013-01-01

    Background During a two year period, a 27-year-old female veterinarian experienced migraine headaches, seizures, including status epilepticus, and other neurological and neurocognitive abnormalities. Prior to and during her illness, she had been actively involved in hospital-based work treating domestic animals, primarily cats and dogs, in Grenada and Ireland and anatomical research requiring the dissection of wild animals (including lions, giraffe, rabbits, mongoose, and other animals), mostly in South Africa. The woman reported contact with fleas, ticks, lice, biting flies, mosquitoes, spiders and mites and had also been scratched or bitten by dogs, cats, birds, horses, reptiles, rabbits and rodents. Prior diagnostic testing resulted in findings that were inconclusive or within normal reference ranges and no etiological diagnosis had been obtained to explain the patient’s symptoms. Methods PCR assays targeting Anaplasma spp. Bartonella spp. and hemotopic Mycoplasma spp. were used to test patient blood samples. PCR positive amplicons were sequenced directly and compared to GenBank sequences. In addition, Bartonella alpha Proteobacteria growth medium (BAPGM) enrichment blood culture was used to facilitate bacterial growth and Bartonella spp. serology was performed by indirect fluorescent antibody testing. Results Anaplasma platys, Bartonella henselae and Candidatus Mycoplasma haematoparvum DNA was amplified and sequenced from the woman’s blood, serum or blood culture samples. Her serum was variably seroreactive to several Bartonella sp. antigens. Despite symptomatic improvement, six months of doxycycline most likely failed to eliminate the B. henselae infection, whereas A. platys and Candidatus M. haematoparvum DNA was no longer amplified from post-treatment samples. Conclusions As is typical of many veterinary professionals, this individual had frequent exposure to arthropod vectors and near daily contact with persistently bacteremic reservoir hosts, including

  17. Seroprevalence and risk factors associated with Ehrlichia canis, Anaplasma spp., Borrelia burgdorferi sensu lato, and D. immitis in hunting dogs from southern Italy.

    PubMed

    Piantedosi, Diego; Neola, Benedetto; D'Alessio, Nicola; Di Prisco, Francesca; Santoro, Mario; Pacifico, Laura; Sgroi, Giovanni; Auletta, Luigi; Buch, Jesse; Chandrashekar, Ramaswamy; Breitschwerdt, Edward B; Veneziano, Vincenzo

    2017-08-04

    Canine vector-borne diseases (CVBDs) are caused by a range of pathogens transmitted to dogs by arthropods. The present study investigates Ehrlichia canis, Anaplasma spp., Borrelia burgdorferi sensu lato, and Dirofilaria immitis seroprevalences in hunting dogs from southern Italy. Dogs (no. 1335) were tested using a commercial in-clinic enzyme-linked immunosorbent assay kit. Odds ratios (ORs) were calculated by logistic regression analysis to identify risk factors. Overall, 138/1335 dogs (10.3%) were seroreactive to at least one CVBD pathogen. E. canis, Anaplasma spp., B. burgdorferi s.l., and D. immitis seroprevalences were 7.6, 4.4, 0.3, and 0.2%, respectively. E. canis and Anaplasma spp. co-exposures were found in 30 dogs (2.2%), compared with Anaplasma spp. and B. burgdorferi s.l. co-exposures in 2 dogs (0.1%). Adult age was a risk factor for E. canis (OR 2.35) seroreactivity whereas hunting fur-bearing animals for E. canis (OR 4.75) and Anaplasma spp. (OR 1.87), respectively. The historical presence of tick infestation was identified as a risk factor for positivity to E. canis (OR 2.08) and Anaplasma spp. (OR 2.15). Finally, larger dog pack size was significantly associated with E. canis (OR 1.85) and Anaplasma spp. (OR 2.42) exposures. The results of the present survey indicated that hunting dog populations are at relative risk of CVBDs in southern Italy. Further studies are needed to evaluate the role of hunting dogs in the epidemiology of vector-borne organisms due to sharing common environments with wild, sympatric animal populations.

  18. Improved diagnostic performance of a commercial anaplasma antibody competitive enzyme-linked immunosorbent assay using recombinant major surface protein 5–glutathione S-transferase fusion protein as antigen

    USDA-ARS?s Scientific Manuscript database

    This study tested the hypothesis that removal of maltose binding protein from recombinant antigen used for plate coating would improve the specificity of Anaplasma antibody competitive ELISA. Three hundred and eight sera with significant MBP antibody binding (=30%I) in Anaplasma negative herds was 1...

  19. Sensory exploitation and sexual conflict

    PubMed Central

    Arnqvist, Göran

    2006-01-01

    Much of the literature on male–female coevolution concerns the processes by which male traits and female preferences for these can coevolve and be maintained by selection. There has been less explicit focus on the origin of male traits and female preferences. Here, I argue that it is important to distinguish origin from subsequent coevolution and that insights into the origin can help us appreciate the relative roles of various coevolutionary processes for the evolution of diversity in sexual dimorphism. I delineate four distinct scenarios for the origin of male traits and female preferences that build on past contributions, two of which are based on pre-existing variation in quality indicators among males and two on exploitation of pre-existing sensory biases among females. Recent empirical research, and theoretical models, suggest that origin by sensory exploitation has been widespread. I argue that this points to a key, but perhaps transient, role for sexually antagonistic coevolution (SAC) in the subsequent evolutionary elaboration of sexual traits, because (i) sensory exploitation is often likely to be initially costly for individuals of the exploited sex and (ii) the subsequent evolution of resistance to sensory exploitation should often be associated with costs due to selective constraints. A review of a few case studies is used to illustrate these points. Empirical data directly relevant to the costs of being sensory exploited and the costs of evolving resistance is largely lacking, and I stress that such data would help determining the general importance of sexual conflict and SAC for the evolution of sexual dimorphism. PMID:16612895

  20. The Exploitation of Black Athletes.

    ERIC Educational Resources Information Center

    Edwards, Harry

    1983-01-01

    Colleges and universities have not up held their end of the bargain with athletes, exploiting a disproportionate number of talented Black athletes by not providing the kind of education the students sought or needed and by applying rigid academic standards for eligibility. (MSE)

  1. Detection of Ehrlichia spp., Anaplasma spp., Rickettsia spp., and other eubacteria in ticks from the Thai-Myanmar border and Vietnam.

    PubMed

    Parola, Philippe; Cornet, Jean-Paul; Sanogo, Yibayiri Osée; Miller, R Scott; Thien, Huynh Van; Gonzalez, Jean-Paul; Raoult, Didier; Telford III, Sam R; Wongsrichanalai, Chansuda

    2003-04-01

    A total of 650 ticks, including 13 species from five genera, were collected from animals, from people, or by flagging of the vegetation at sites on the Thai-Myanmar border and in Vietnam. They were tested by PCR to detect DNA of bacteria of the order RICKETTSIALES: Three Anaplasma spp. were detected in ticks collected in Thailand, including (i) Anaplasma sp. strain AnDa465, which was considered a genotype of Anaplasma platys (formerly Ehrlichia platys) and which was obtained from Dermacentor auratus ticks collected from dogs; (ii) Anaplasma sp. strain AnAj360, which was obtained from Amblyomma javanense ticks collected on a pangolin; and (iii) Anaplasma sp. strain AnHl446, which was closely related to Anaplasma bovis and which was detected in Haemaphysalis lagrangei ticks collected from a bear. Three Ehrlichia spp. were identified, including (i) Ehrlichia sp. strain EBm52, which was obtained from Boophilus microplus ticks collected from cattle from Thailand; (ii) Ehrlichia sp. strain EHh324, which was closely related to Ehrlichia chaffeensis and which was detected in Haemaphysalis hystricis ticks collected from wild pigs in Vietnam; and (iii) Ehrlichia sp. strain EHh317, which was closely related to Ehrlichia sp. strain EBm52 and which was also detected in H. hystricis ticks collected from wild pigs in Vietnam. Two Rickettsia spp. were detected in Thailand, including (i) Rickettsia sp. strain RDla420, which was detected in Dermacentor auratus ticks collected from a bear, and (ii) Rickettsia sp. strain RDla440, which was identified from two pools of Dermacentor larvae collected from a wild pig nest. Finally, two bacteria named Eubacterium sp. strain Hw124 and Eubacterium sp. strain Hw191 were identified in Haemaphysalis wellingtoni ticks collected from chicken in Thailand; these strains could belong to a new group of bacteria.

  2. The occurrence of Dirofilaria immitis, Borrelia burgdorferi, Ehrlichia canis and Anaplasma phagocytophium in dogs in China.

    PubMed

    Xia, Z; Yu, D; Mao, J; Zhang, Z; Yu, J

    2012-06-01

    A survey of the occurrence of Dirofilaria immitis, Borrelia burgdorferi, Ehrlichia canis and Anaplasma phagocytophium in dogs was undertaken in the People's Republic of China between October 2008 and October 2009. A total of 600 blood samples were taken from dogs in four cities in China: 300 in Beijing, 150 in Shenzhen, 30 in Shanghai and 120 in Zhengzhou. All samples were tested for the heartworm antigen and antibodies of canine B. burgdorferi, E. canis and A. phagocytophium by using the canine SNAP® 4Dx® test kit. The occurrence of D. immitis, B. burgdorferi, E. canis and A. phagocytophium was 1.17% (7/600), 0.17% (1/600), 2.17% (13/600) and 0.5% (3/600), respectively. In Shenzhen city 2% (3/150), 8.67% (13/150) and 2% (3/150) of samples were positive for D. immitis, E. canis and A. phagocytophium, respectively. The occurrence of heartworm antigen was 0.33% (1/300) in Beijing, 2.00% (3/150) in Shenzhen, 3.33% (1/30) in Shanghai and 1.67% (2/120) in Zhengzhou. We found E. canis and A. phagocytophium only at one site, Shenzhen, while the only occurrence of B. burgdorferi was at Beijing. In conclusion, the dog population in China is at potential risk for D. immitis, B. burgdorferi, E. canis and A. phagocytophium infection, the risk being especially high in southern China.

  3. Occurrence of Ehrlichia canis and Anaplasma platys in household dogs from northern Parana.

    PubMed

    da Silva, Gislaine Cristina Ferreira; Benitez, Aline do Nascimento; Girotto, Aline; Taroda, Alessandra; Vidotto, Marilda Carlos; Garcia, João Luis; de Freitas, Julio Cesar; Arlington, Selwyn Headley; Vidotto, Odilon

    2012-01-01

    Canine monocytic ehrlichiosis caused primarily by Ehrlichia canis and canine thrombocytic anaplasmosis induced by Anaplasma platys are important emerging zoonotic tick-borne diseases of dogs. There is evidence that these pathogens can also affect humans. This study evaluated the presence of E. canis and A. platys in blood samples collected from 256 domiciled dogs in the municipality of Jataizinho, located in north region of the State of Parana, Brazil, by PCR assay. The occurrence of E. canis and A. platys was 16.4% (42/256) and 19.4% (49/256), respectively; while 5.47% (14/256) of the dogs evaluated were co-infected by these two organisms. The presence of E. canis and A. platys was not significantly associated with the variables evaluated (sex, age, outdoor access, and presence of ticks during blood collection). Infection of dogs by E. canis was associated with anemia and thrombocytopenia, while infection induced by A. platys was related only to thrombocytopenia. Canine monocytic ehrlichiosis and canine thrombocytic anaplasmosis should be included in the differential diagnoses when these hematological alterations are observed during routine laboratory evaluation of dogs.

  4. Detection of Ehrlichia canis and Anaplasma platys DNA using multiplex PCR.

    PubMed

    Rufino, Claudia Pinheiro; Moraes, Pablo Henrique Gonçalves; Reis, Thais; Campos, Ruan; Aguiar, Délia Cristina Figueira; McCulloch, John Anthony; Meneses, Andre Marcelo Conceição; Gonçalves, Evonnildo Costa

    2013-12-01

    We hereby propose a novel sensitive, specific, and cost-efficient method to detect Ehrlichia canis and Anaplasma platys DNA from canine whole blood samples by multiplex PCR. Blood samples from hemoparasited dogs attending the Veterinary Hospital at the Universidade Federal Rural da Amazônia-UFRA, Belém, Brazil, were collected in tubes containing EDTA. Amplification of E. canis and A. platys 16S rDNA by nested (n) PCR was successfully achieved by using primers specific to the Anaplasmataceae in the first round of PCR, followed by a second round of PCR using E. canis-specific primers in conjunction with A. platys-specific primers. The amplicons obtained were cloned and sequenced, yielding sequences of 478 and 473 bp (including primers) pertaining to regions of the 16S rDNA of E. canis and A. platys, respectively. The protocol we here propose may help to measure the prevalence of canine monocytic ehrlichiosis (CME) and canine cyclic thrompocytopenia, not only in northern Brazil, where there is no data available, but also elsewhere.

  5. Molecular detection of Anaplasma platys and Ehrlichia canis in dogs from Kabylie, Algeria.

    PubMed

    Dahmani, Mustapha; Loudahi, Abdelghani; Mediannikov, Oleg; Fenollar, Florence; Raoult, Didier; Davoust, Bernard

    2015-03-01

    Ehrlichia canis and Anaplasma platys are bacteria belonging to the Anaplasmataceae family that cause acute, self-limiting and sometimes fatal vector-borne infections in dogs. These bacteria have been reported worldwide and are transmitted mainly by Rhipicephalus sanguineus. Aside from a report on E. canis once in 1935, no other Anaplasmataceae bacteria have been reported in Algeria to date. The aim of this study was to identify the microbial species implicated in ehrlichiosis and anaplasmosis by a molecular epidemiological survey in dogs. The study was carried out in Kabylie, in northeast Algeria. Sampling was performed in 11 municipalities in the province of Tizi Ouzou and 2 municipalities in the province of Béjaïa. Peripheral blood samples from 110 dogs were screened by qPCR, which is capable of identifying most Anaplasmataceae bacteria. Out of 110, a total of 13 samples screened positive (7/110 E. canis and 6/110 A. platys), and two genetic variants of A. platys and one of E. canis were identified. This is the first study to report the presence of A. platys in dogs from Algeria using a molecular investigative method. This survey was conducted in early spring. As tick activity can affect the prevalence of these pathogens in dogs, further investigations are needed to establish the year-round prevalence of these infections.

  6. Use of Percoll gradients to purify Anaplasma marginale (Rickettsiales: Anaplasmataceae) from tick cell cultures.

    PubMed

    Lis, Katarzyna; Najm, Nour; de la Fuente, José; Fernández de Mera, Isabel; Zweygarth, Erich; Pfister, Kurt; Passos, Lygia M F

    2014-09-01

    Anaplasma marginale (Rickettsiales: Anaplasmataceae) is an obligate intracellular bacterium that multiplies exclusively within membrane-bound vacuoles in the cytoplasm of host cells. A number of A. marginale isolates can be propagated in the Ixodes scapularis IDE8 tick cell line, which provides a reliable source of antigens for a wide variety of studies. However, because of its intracellular nature, separation of bacteria from host cell materials remains an important constraint for researchers. In the present study, we evaluated the use of Percoll gradients for purification of two Brazilian strains of A. marginale grown in IDE8 tick cells. The purified A. marginale monitored in Giemsa-stained smears contained only minimal amounts of IDE8 cell stroma. The total protein yields were 1.2mg and 1.7mg, while the DNA titers quantified with real-time PCR were 6.4×10(9) for UFMG1 and 4.87×10(9) for UFMG2 copies in the purified material, respectively. Additionally, we confirmed the viability of purified bacteria by infecting tick cells after being freshly purified and after retrieval from long-term storage. Importantly, the viability of the organisms is preserved after use of this separation method, and therefore the purified organisms can be used in enzymatic assays and other research approaches where live organisms would be preferred. Copyright © 2014 Elsevier GmbH. All rights reserved.

  7. Detection and phylogenetic characterization of Theileria spp. and Anaplasma marginale in Rhipicephalus bursa in Portugal.

    PubMed

    Ferrolho, Joana; Antunes, Sandra; Santos, Ana S; Velez, Rita; Padre, Ludovina; Cabezas-Cruz, Alejandro; Santos-Silva, Maria Margarida; Domingos, Ana

    2016-04-01

    Ticks are obligatory blood-sucking arthropod (Acari:Ixodida) ectoparasites of domestic and wild animals as well as humans. The incidence of tick-borne diseases is rising worldwide, challenging our approach toward diagnosis, treatment and control options. Rhipicephalus bursa Canestrini and Fanzago, 1877, a two-host tick widely distributed in the Palearctic Mediterranean region, is considered a multi-host tick that can be commonly found on sheep, goats and cattle, and occasionally on horses, dogs, deer and humans. R. bursa is a species involved in the transmission of several tick-borne pathogens with a known impact on animal health and production. The aim of this study was to estimate R. bursa prevalence in Portugal Mainland and circulating pathogens in order to contribute to a better knowledge of the impact of this tick species. Anaplasma marginale and Theileria spp. were detected and classified using phylogenetic analysis. This is the first report of Theileria annulata and Theileria equi detection in R. bursa ticks feeding on cattle and horses, respectively, in Portugal. This study contributes toward the identification of currently circulating pathogens in this tick species as a prerequisite for developing future effective anti-tick control measures.

  8. Functional and Immunological Relevance of Anaplasma marginale Major Surface Protein 1a Sequence and Structural Analysis

    PubMed Central

    Cabezas-Cruz, Alejandro; Passos, Lygia M. F.; Lis, Katarzyna; Kenneil, Rachel; Valdés, James J.; Ferrolho, Joana; Tonk, Miray; Pohl, Anna E.; Grubhoffer, Libor; Zweygarth, Erich; Shkap, Varda; Ribeiro, Mucio F. B.; Estrada-Peña, Agustín; Kocan, Katherine M.; de la Fuente, José

    2013-01-01

    Bovine anaplasmosis is caused by cattle infection with the tick-borne bacterium, Anaplasma marginale. The major surface protein 1a (MSP1a) has been used as a genetic marker for identifying A. marginale strains based on N-terminal tandem repeats and a 5′-UTR microsatellite located in the msp1a gene. The MSP1a tandem repeats contain immune relevant elements and functional domains that bind to bovine erythrocytes and tick cells, thus providing information about the evolution of host-pathogen and vector-pathogen interactions. Here we propose one nomenclature for A. marginale strain classification based on MSP1a. All tandem repeats among A. marginale strains were classified and the amino acid variability/frequency in each position was determined. The sequence variation at immunodominant B cell epitopes was determined and the secondary (2D) structure of the tandem repeats was modeled. A total of 224 different strains of A. marginale were classified, showing 11 genotypes based on the 5′-UTR microsatellite and 193 different tandem repeats with high amino acid variability per position. Our results showed phylogenetic correlation between MSP1a sequence, secondary structure, B-cell epitope composition and tick transmissibility of A. marginale strains. The analysis of MSP1a sequences provides relevant information about the biology of A. marginale to design vaccines with a cross-protective capacity based on MSP1a B-cell epitopes. PMID:23776456

  9. Comparison of proteins synthesized by two different isolates of Anaplasma marginale.

    PubMed Central

    Barbet, A F; Anderson, L W; Palmer, G H; McGuire, T C

    1983-01-01

    We present results on the initial definition of proteins synthesized by two isolates of Anaplasma marginale. Bovine erythrocytes infected with A. marginale were radioactively labeled with [35S]methionine or a 3H-amino acid mixture during short-term in vitro culture. The labeled proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This technique revealed protein bands of various apparent molecular weights from less than 14,000 to greater than 200,000. The bands observed represented A. marginale proteins because (i) uninfected erythrocytes from the same animal did not incorporate radioisotope during identical culture conditions, and (ii) the incorporation of radioisotope into proteins during culture of infected erythrocytes was inhibited by tetracycline but not by cycloheximide. The radioactive protein profiles of two different isolates of A. marginale, from Washington and Florida, were compared by two-dimensional gel electrophoresis. About 200 proteins were resolved in each case. Several proteins differed in position when the two-dimensional gel maps were compared, indicating variations in protein structure between the two A. marginale isolates. Images PMID:6852911

  10. Protective immunity induced by immunization with a live, cultured Anaplasma marginale strain

    PubMed Central

    Hammac, G. Kenitra; Ku, Pei-Shin; Galletti, Maria F.; Noh, Susan M.; Scoles, Glen A.; Palmer, Guy H.; Brayton, Kelly A.

    2014-01-01

    Despite significant economic losses resulting from infection with Anaplasma marginale, a tick-transmitted rickettsial pathogen of cattle, available vaccines provide, at best, only partial protection against clinical disease. The green-fluorescent protein expressing mutant of the A. marginale St. Maries strain is a live, marked vaccine candidate (AmStM-GFP1). To test whether AmStM-GFP is safe and provides clinical protection, a group of calves was vaccinated, and clinical parameters, including percent parasitized erythrocytes (PPE), packed cell volume (PCV) and days required to reach peak bacteremia, were measured following inoculation and following tick challenge with wild type St Maries strain (AmStM). These clinical parameters were compared to those obtained during infection with the A. marginale subsp. centrale vaccine strain (A. centrale) or wild type AmStM. AmStM-GFP resulted in similar clinical parameters to A. centrale, but had a lower maximum PPE, smaller drop in PCV and took longer to reach peak bacteremia than wild type AmStM. AmStM-GFP provided clinical protection, yielding a stable PCV and low bacteremia following challenge, whereas A. centrale only afforded partial clinical protection. PMID:23664994

  11. Association of Anaplasma marginale Strain Superinfection with Infection Prevalence within Tropical Regions

    PubMed Central

    Castañeda-Ortiz, Elizabeth J.; Ueti, Massaro W.; Camacho-Nuez, Minerva; Mosqueda, Juan J.; Mousel, Michelle R.; Johnson, Wendell C.; Palmer, Guy H.

    2015-01-01

    Strain superinfection occurs when a second strain infects a host already infected with and having mounted an immune response to a primary strain. The incidence of superinfection with Anaplasma marginale, a tick-borne rickettsial pathogen of domestic and wild ruminants, has been shown to be higher in tropical versus temperate regions. This has been attributed to the higher prevalence of infection, with consequent immunity against primary strains and thus greater selective pressure for superinfection with antigenically distinct strains. However an alternative explanation would be the differences in the transmitting vector, Dermacentor andersoni in the studied temperate regions and Rhipicephalus microplus in the studied tropical regions. To address this question, we examined two tropical populations sharing the same vector, R. microplus, but with significantly different infection prevalence. Using two separate markers, msp1α (one allele per genome) and msp2 (multiple alleles per genome), there were higher levels of multiple strain infections in the high infection prevalence as compared to the low prevalence population. The association of higher strain diversity with infection prevalence supports the hypothesis that high levels of infection prevalence and consequent population immunity is the predominant driver of strain superinfection. PMID:25793966

  12. Dark matters: exploitation as cooperation.

    PubMed

    Dasgupta, Partha

    2012-04-21

    The empirical literature on human cooperation contains studies of communitarian institutions that govern the provision of public goods and management of common property resources in poor countries. Scholars studying those institutions have frequently used the Prisoners' Dilemma game as their theoretical tool-kit. But neither the provision of local public goods nor the management of local common property resources involves the Prisoners' Dilemma. That has implications for our reading of communitarian institutions. By applying a fundamental result in the theory of repeated games to a model of local common property resources, it is shown that communitarian institutions can harbour exploitation of fellow members, something that would not be possible in societies where cooperation amounts to overcoming the Prisoners' Dilemma. The conclusion we should draw is that exploitation can masquerade as cooperation.

  13. Teotihuacan, tepeapulco, and obsidian exploitation.

    PubMed

    Charlton, T H

    1978-06-16

    Current cultural ecological models of the development of civilization in central Mexico emphasize the role of subsistence production techniques and organization. The recent use of established and productive archeological surface survey techniques along natural corridors of communication between favorable niches for cultural development within the Central Mexican symbiotic region resulted in the location of sites that indicate an early development of a decentralized resource exploitation, manufacturing, and exchange network. The association of the development of this system with Teotihuacán indicates the importance such nonsubsistence production and exchange had in the evolution of this first central Mexican civilization. The later expansion of Teotihuacán into more distant areas of Mesoamerica was based on this resource exploitation model. Later civilizations centered at Tula and Tenochtitlán also used such a model in their expansion.

  14. Canine infection with Dirofilaria immitis, Borrelia burgdorferi, Anaplasma spp., and Ehrlichia spp. in the United States, 2010–2012

    PubMed Central

    2014-01-01

    Background The geographic distribution of canine infection with vector-borne disease agents in the United States appears to be expanding. Methods To provide an updated assessment of geographic trends in canine infection with Dirofilaria immitis, Borrelia burgdorferi, Ehrlichia spp., and Anaplasma spp., we evaluated results from an average of 3,588,477 dogs tested annually by veterinarians throughout the United States from 2010 – 2012. Results As in an earlier summary report, the percent positive test results varied by agent and region, with antigen of D. immitis and antibody to Ehrlichia spp. most commonly identified in the Southeast (2.9% and 3.2%, respectively) and antibody to both B. burgdorferi and Anaplasma spp. most commonly identified in the Northeast (13.3% and 7.1%, respectively) and upper Midwest (4.4% and 3.9%, respectively). Percent positive test results for D. immitis antigen were lower in every region considered, including in the Southeast, than previously reported. Percent positive test results for antibodies to B. burgdorferi and Ehrlichia spp. were higher nationally than previously reported, and, for antibodies to Anaplasma spp., were higher in the Northeast but lower in the Midwest and West, than in the initial report. Annual reports of human cases of Lyme disease, ehrlichiosis, and anaplasmosis were associated with percent positive canine test results by state for each respective tick-borne disease agent (R2 = 0.701, 0.457, and 0.314, respectively). Within endemic areas, percent positive test results for all three tick-borne agents demonstrated evidence of geographic expansion. Conclusions Continued national monitoring of canine test results for vector-borne zoonotic agents is an important tool for accurately mapping the geographic distribution of these agents, and greatly aids our understanding of the veterinary and public health threats they pose. PMID:24886589

  15. Routing Algorithm Exploits Spatial Relations

    NASA Technical Reports Server (NTRS)

    Okino, Clayton; Jennings, Esther

    2004-01-01

    A recently developed routing algorithm for broadcasting in an ad hoc wireless communication network takes account of, and exploits, the spatial relationships among the locations of nodes, in addition to transmission power levels and distances between the nodes. In contrast, most prior algorithms for discovering routes through ad hoc networks rely heavily on transmission power levels and utilize limited graph-topology techniques that do not involve consideration of the aforesaid spatial relationships. The present algorithm extracts the relevant spatial-relationship information by use of a construct denoted the relative-neighborhood graph (RNG).

  16. Immunization-induced anaplasma marginale-specific T lymphocyte reponses impaired by A. marginale infection are restored after eliminating the infection with tetracycline

    USDA-ARS?s Scientific Manuscript database

    Infection of cattle with Anaplasma marginale fails to prime sustained effector/memory T-cell responses, and high bacterial load may induce antigen-specific CD4 T exhaustion and deletion. We tested the hypothesis that clearance of persistent infection restores the exhausted T-cell response. We show t...

  17. Anaplasma marginale infection with persistent high-load bacteremia induces a dysfunctional memory CD4+ T lymphocyte response but sustained high IgG titers

    USDA-ARS?s Scientific Manuscript database

    Control of blood-borne infections is dependent on antigen-specific effector and memory T cells and high-affinity IgG responses. In chronic infections characterized by a high antigen load, it has been shown that antigen-specific T and B cells are vulnerable to downregulation and apoptosis. Anaplasma ...

  18. Molecular and serological in-herd prevalence of Anaplasma marginale infection in Texas cattle.

    PubMed

    Hairgrove, Thomas; Schroeder, Megan E; Budke, Christine M; Rodgers, Sandy; Chung, Chungwon; Ueti, Massaro W; Bounpheng, Mangkey A

    2015-04-01

    Bovine anaplasmosis is an infectious, non-contagious disease caused by the rickettsial pathogen Anaplasma marginale (A. marginale). The organism has a global distribution and infects erythrocytes, resulting in anemia, jaundice, fever, abortions and death. Once infected, animals remain carriers for life. The carrier status provides immunity to clinical disease, but is problematic if infected and naïve cattle are comingled. Knowledge of infection prevalence and spatial distribution is important in disease management. The objective of this study was to assess A. marginale infection in-herd prevalence in Texas cattle using both molecular and serological methods. Blood samples from 11 cattle herds within Texas were collected and analyzed by reverse transcription quantitative real-time PCR (RT-qPCR) and a commercial competitive enzyme-linked immunosorbent assay (cELISA). Samples from experimentally infected animals were also analyzed and RT-qPCR detected A. marginale infection up to 15 days before cELISA, providing empirical data to support the interpretation of herd prevalence results. Herds with high prevalence were located in the north Texas Rolling Plains and west Trans-Pecos Desert, with RT-qPCR prevalence as high as 82% and cELISA prevalence as high as 88%. Overall prevalence was significantly higher in cattle in north and west Texas compared to cattle in east Texas (p<0.0001 for prevalence based on both RT-qPCR and cELISA). The overall RT-qPCR and cELISA results exhibited 90% agreement (kappa=0.79) and provide the first A. marginale infection prevalence study for Texas cattle using two diagnostic methods. Since cattle are the most important reservoir host for A. marginale and can serve as a source of infection for tick and mechanical transmission, information on infection prevalence is beneficial in the development of prevention and control strategies. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Genetic diversity of Anaplasma marginale strains from an outbreak of bovine anaplasmosis in an endemic area.

    PubMed

    Almazán, Consuelo; Alamzán, Consuelo; Medrano, Citlaly; Ortiz, Martín; de la Fuente, José

    2008-11-25

    Anaplasma marginale is a tick-borne pathogen of cattle that causes the disease bovine anaplasmosis worldwide. Major surface proteins (MSPs) are involved in host-pathogen and tick-pathogen interactions and have been used as markers for the genetic characterization of A. marginale strains. A. marginale genotypes are highly variable in endemic areas worldwide. The genetic composition of A. marginale strains during anaplasmosis outbreaks has been characterized in one study only which reported a single msp1alpha genotype in infected cattle. However, more information is required to characterize whether a single genotype is responsible for an anaplasmosis outbreak or whether multiple genotypes can cause disease in naïve cattle within a single herd in endemic areas. The aim of this study was to characterize the genetic diversity of A. marginale strains from an outbreak of bovine anaplasmosis in the State of Tamaulipas, Mexico. A. marginale genotypes were characterized at the molecular level using msp4 and msp1alpha gene sequences. The results revealed that several A. marginale genotypes are present in cattle during acute anaplasmosis outbreaks, thus suggesting that mechanical transmission or stochastic biological transmission through equally efficient independent transmission events may explain A. marginale genotype frequency in a cattle herd during acute bovine anaplasmosis outbreaks in endemic areas. The results reported herein corroborated the genetic heterogeneity of A. marginale strains in endemic regions worldwide. The development and implementation of anaplasmosis control measures is dependent upon understanding the epidemiology of A. marginale in endemic regions, including the characterization of the genetic diversity of strains that produce outbreaks of bovine anaplasmosis.

  20. Molecular characterization of Anaplasma marginale in ticks naturally feeding on buffaloes.

    PubMed

    da Silva, Jenevaldo Barbosa; da Fonseca, Adivaldo Henrique; Barbosa, José Diomedes

    2015-10-01

    Anaplasma marginale is the most prevalent pathogen transmitted by ticks in cattle in tropical and subtropical regions of the world. However, the tick species involved in the transmission of A. marginale in buffaloes in Brazil have not been identified. The objective of the present study was to determine the presence of A. marginale in ticks parasitizing water buffaloes. A total of 200 samples of Rhipicephalus microplus, Dermacentor nitens, Amblyomma cajennense, and Amblyomma maculatum were collected and tested by conventional and quantitative PCR for the presence of the msp1a and msp5 genes. In the present study, 35 ticks (17.5%) were positive for A. marginale DNA by qPCR analysis. The positive ticks belonged to four different species: R. microplus (22.2%), A. cajennense (13.8%), A. maculatum (16.0%), and D. nitens (10.0%). Individuals of the three developmental stages (larvae, nymphs, and adults) of R. microplus and A. cajennense were found to be positive for A. marginale, only nymphs and adults of A. maculatum were found to be positive, and finally, only adults of D. nitens were positive for A. marginale. Our results suggest that R. microplus, A. cajennense, A. maculatum, and D. nitens ticks may be involved in the transmission of A. marginale in buffaloes. However, while A. marginale PCR positive ticks were recorded, this does not indicate vector competence; only that the ticks may contain a blood meal from an infected host. Additionally, the results show that the strains of A. marginale from buffaloes and cattle are phylogenetically related.

  1. Seroprevalence of Anaplasma marginale and Babesia bigemina infections and associated risk factors in Machakos County, Kenya.

    PubMed

    Wesonga, F D; Gachohi, J M; Kitala, P M; Gathuma, J M; Njenga, M J

    2017-02-01

    Anaplasma marginale and Babesia bigemina are important tick-borne pathogens of cattle. A cross-sectional survey was undertaken to determine the seroprevalence of A. marginale and B. bigemina infections and identify associated risk factors on traditional smallholder farms in Machakos County, Kenya. A total of 421 cattle from 127 farms from four divisions in the county were sampled and visited between September and November 2007. The farms were selected by a proportional allocation approach based on the number of farms in the four divisions previously selected by stratified random sampling method. Information on animal and individual farm management variables was obtained using standardized questionnaires. Prevalence of serum antibodies due to A. marginale and B. bigemina pathogens was determined using the enzyme-linked immunosorbent assay (ELISA) technique. The relationship between the seropositivity and associated risk factors was assessed by multivariable analyses using standard logistic regression models. The overall estimation (and their 95% confidence intervals) of A. marginale and B. bigemina seropositivity at the animal level was 53.4% (48.5%, 58.2%) and 40.6% (35.8%, 45.4%), respectively. Two variables, "animal age" and "administrative division," were significantly associated with the A. marginale seroresponse. Three variables, "animal age" "grazing system" and "administrative division" were significantly associated with the B. bigemina seroresponse. These findings suggest possible indicators of existence of endemic instability for the two infections. The study identifies characterization of environmental suitability for the vectors and how they interact with grazing systems to cause the infections as an area for further studies, for improved understanding of the infections and in designing disease control programs.

  2. Intraoperative bleeding in dogs from Grenada seroreactive to Anaplasma platys and Ehrlichia canis.

    PubMed

    Lanza-Perea, M; Zieger, U; Qurollo, B A; Hegarty, B C; Pultorak, E L; Kumthekar, S; Bruhl-Day, R; Breitschwerdt, E B

    2014-01-01

    Frequent exposure of Grenadian dogs to Rhipicephalus sanguineus results in Anaplasma platys, and Ehrlichia canis seroreactivity. During elective surgeries, substantial intraoperative hemorrhage occurs in some seroreactive dogs. To assess hemostatic parameters and bleeding tendencies as well as prevalence of PCR positivity in apparently healthy A. platys and E. canis seroreactive and seronegative free-roaming dogs from Grenada. Forty-seven elective surgery dogs allocated to 4 groups: Seronegative control (n = 12), A. platys (n = 10), E. canis (n = 14) and A. platys, and E. canis (n = 11) seroreactive. Preoperatively, hemostasis was assessed by platelet count, prothrombin time, activated partial thromboplastin time, and buccal mucosal bleeding time. Intra- and postoperative bleeding scores were subjectively assigned. Blood, spleen, bone marrow, and lymph node aspirates were tested by PCR. Bleeding scores in dogs coseroreactive for A. platys and E. canis were higher (P = .015) than those of seronegative dogs. A. platys DNA was amplified from 7/21 (33%) A. platys seroreactive dogs and from 1 E. canis seroreactive dog; E. canis DNA was amplified from 21/25 (84%) E. canis seroreactive dogs. E. canis DNA was amplified most often from blood, whereas A. platys DNA was amplified most often from bone marrow. Apparently healthy, free-roaming dogs coseropositive for A. platys and E. canis may have increased intraoperative bleeding tendencies despite normal hemostatic parameters. Future investigations should explore the potential for vascular injury as a cause for bleeding in these dogs. Improved tick control is needed for dogs in Grenada. Copyright © 2014 by the American College of Veterinary Internal Medicine.

  3. Prevalence and haemato-biochemical profile of Anaplasma marginale infection in dairy animals of Punjab (India).

    PubMed

    Ashuma; Sharma, Amrita; Singla, Lachhman Das; Kaur, Paramjit; Bal, Mandeep Singh; Batth, Balwinder Kaur; Juyal, Prayag Dutt

    2013-02-01

    To do the systematic comparison of prevalence of anaplasmosis by PCR and Giemsa stained thin blood smear (GSTBS) based parasitological assays in dairy cattle of Punjab, which has not been reported yet. To analyse the haematobiochemical alterations in infected animals to arrive at the conclusion regarding the pathogenicity induced by Anaplasma marginale (A. marginale) in latent and patent infection. Study was conducted on 320 animals (236 cows, 62 calves and 22 buffaloes) of Punjab, India. PCR on genome of A. marginale was performed by targeting msp1 β gene using specific primers BAP-2/AL34S, amplifies products of size 407 bp. Questionnaires based data on the characteristics of the infected animals and management strategies of the farm were collected and correlated. Higher prevalence and more significant association was observed in the PCR based molecular diagnosis (P=0.00012) as compared to that in GSTBS (P=0.028 8) based diagnosis with various regions under study. With respect to the regions, highest prevalence was recorded in Ferozepur by PCR based diagnosis, while that in Jalandhar by GSTBS examination. Similar marked significant association of the PCR based diagnosis with the age of the animals under study (P=0.00013) was observed elucidating no inverse age resistance to A. marginale in cow calves. Haematobiochemical profile of infected animals revealed marked anemia, liver dysfunction and increase globulin concentrate indicating rise in immunoglobulin level to counteract infection. PCR is far more sensitive in detecting the disease even in latent infection which may act as nidus for spread of anaplasmosis to susceptible animals in endemic areas. Severity of anaemia and liver dysfunction were comparable both in patent as well as latent infection indicating pathogenicity of both. Copyright © 2013 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

  4. Global transcriptional analysis reveals surface remodeling of Anaplasma marginale in the tick vector

    PubMed Central

    2014-01-01

    Background Pathogens dependent upon vectors for transmission to new hosts undergo environment specific changes in gene transcription dependent on whether they are replicating in the vector or the mammalian host. Differential gene transcription, especially of potential vaccine candidates, is of interest in Anaplasma marginale, the tick-borne causative agent of bovine anaplasmosis. Methods RNA-seq technology allowed a comprehensive analysis of the transcriptional status of A. marginale genes in two conditions: bovine host blood and tick derived cell culture, a model for the tick vector. Quantitative PCR was used to assess transcription of a set of genes in A. marginale infected tick midguts and salivary glands at two time points during the transmission cycle. Results Genes belonging to fourteen pathways or component groups were found to be differentially transcribed in A. marginale in the bovine host versus the tick vector. One of the most significantly altered groups was composed of surface proteins. Of the 56 genes included in the surface protein group, eight were up regulated and 26 were down regulated. The down regulated surface protein encoding genes include several that are well studied due to their immunogenicity and function. Quantitative PCR of a set of genes demonstrated that transcription in tick cell culture most closely approximates transcription in salivary glands of recently infected ticks. Conclusions The ISE6 tick cell culture line is an acceptable model for early infection in tick salivary glands, and reveals disproportionate down regulation of surface protein genes in the tick. Transcriptional profiling in other cell lines may help us simulate additional microenvironments. Understanding vector-specific alteration of gene transcription, especially of surface protein encoding genes, may aid in the development of vaccines or transmission blocking therapies. PMID:24751137

  5. Structural Basis for Recombinatorial Permissiveness in the Generation of Anaplasma marginale Msp2 Antigenic Variants

    PubMed Central

    Silva, Marta G.; Kostyukova, Alla S.; Palmer, Guy H.

    2016-01-01

    Sequential expression of outer membrane protein antigenic variants is an evolutionarily convergent mechanism used by bacterial pathogens to escape host immune clearance and establish persistent infection. Variants must be sufficiently structurally distinct to escape existing immune effectors yet retain the core structural elements required for localization and function within the outer membrane. We examined this balance using Anaplasma marginale, which generates antigenic variants in the outer membrane protein Msp2 using gene conversion. The overwhelming majority of Msp2 variants expressed during long-term persistent infection are mosaics, derived by recombination of oligonucleotide segments from multiple alleles to form unique hypervariable regions (HVR). As a result, the mosaics are not under long-term selective pressure to encode a functional protein; consequently, we hypothesized that the Msp2 HVR is structurally permissive for mosaic expression. Using an integrated approach of predictive modeling with determination of the native Msp2 protein structure and function, we demonstrate that structured elements, most notably, β-sheets, are significantly concentrated in the highly conserved N- and C-terminal domains. In contrast, the HVR is overwhelmingly a random coil, with the structured α-helices and β-sheets being confined to the genomically defined structural tethers that separate the antigenically variable microdomains. This structure is supported by the surface exposure of the HVR microdomains and the slow diffusion-type porin function in native Msp2. Importantly, the predominance of the random coil provides plasticity for the formation of functional HVR mosaics and realization of the full potential of segmental gene conversion to dramatically expand the variant repertoire. PMID:27400719

  6. The efficacy of three chlortetracycline regimens in the treatment of persistent Anaplasma marginale infection.

    PubMed

    Reinbold, James B; Coetzee, Johann F; Hollis, Larry C; Nickell, Jason S; Riegel, Casey; Olson, K C; Ganta, Roman R

    2010-09-28

    Chemosterilization is reported in cattle fed chlortetracycline hydrochloride (CTC) at dosages ranging from 1.1mg/kg for 120 days to 11 mg/kg for 30-60 days. The relationship between plasma CTC drug concentration and carrier clearance has not been described. Chronic carrier status was established in 21 steers with a Virginia isolate of Anaplasma marginale and confirmed by cELISA and an A. marginale-specific RT-PCR. Four negative, splenectomized steers served as active disease transmission sentinels. Steers were randomized to receive 4.4 mg/kg/day (LD); 11 mg/kg/day (MD); or 22 mg/kg/day (HD) of oral chlortetracycline; or placebo (CONTROL) for 80 days. The LD, MD and HD treatment groups consisted of 5 infected steers and 1 splenectomized steer; CONTROL group had six infected steers and 1 splenectomized steer. The daily treatments and ration were divided equally and fed twice daily. Blood samples were collected semi-weekly for determining plasma drug concentration by ultrahigh performance liquid chromatography-mass spectrometry/mass spectrometry method and assessment of disease status by both cELISA and RT-PCR. Mean (CV%) chlortetracycline plasma drug concentrations in the LD, MD, and HD groups were 85.3 (28%), 214.5 (32%) and 518.9 (40%)ng/mL during days 4 through 53 of treatment. A negative RT-PCR assay result was confirmed in all CTC-treated groups within 49 days of treatment; however, cELISA required an additional 49 to 88 days before similar results. Subinoculation of splenectomized steers confirmed chemosterilization. These results are important for influencing future chemosterilization strategies and impacting free trade policy among countries and regions of contrasting endemicity. (c) 2010 Elsevier B.V. All rights reserved.

  7. Identification of Midgut and Salivary Glands as Specific and Distinct Barriers to Efficient Tick-Borne Transmission of Anaplasma marginale▿

    PubMed Central

    Ueti, Massaro W.; Reagan, James O.; Knowles, Donald P.; Scoles, Glen A.; Shkap, Varda; Palmer, Guy H.

    2007-01-01

    Understanding the determinants of efficient tick-borne microbial transmission is needed to better predict the emergence of highly transmissible pathogen strains and disease outbreaks. Although the basic developmental cycle of Anaplasma and Ehrlichia spp. within the tick has been delineated, there are marked differences in the ability of specific strains to be efficiently tick transmitted. Using the highly transmissible St. Maries strain of Anaplasma marginale in Dermacentor andersoni as a positive control and two unrelated nontransmissible strains, we identified distinct barriers to efficient transmission within the tick. The Mississippi strain was unable to establish infection at the level of the midgut epithelium despite successful ingestion of infected blood following acquisition feeding on a bacteremic animal host. This inability to colonize the midgut epithelium prevented subsequent development within the salivary glands and transmission. In contrast, A. marginale subsp. centrale colonized the midgut and then the salivary glands, replicating to a titer indistinguishable from that of the highly transmissible St. Maries strain and at least 100 times greater than that previously associated with successful transmission. Nonetheless, A. marginale subsp. centrale was not transmitted, even when a large number of infected ticks was used for transmission feeding. These results establish that there are at least two specific barriers to efficient tick-borne transmission, the midgut and salivary glands, and highlight the complexity of the pathogen-tick interaction. PMID:17420231

  8. Surveillance of Egyptian fleas for agents of public health significance: Anaplasma, Bartonella, Coxiella, Ehrlichia, Rickettsia, and Yersinia pestis.

    PubMed

    Loftis, Amanda D; Reeves, Will K; Szumlas, Daniel E; Abbassy, Magda M; Helmy, Ibrahim M; Moriarity, John R; Dasch, Gregory A

    2006-07-01

    Serologic surveys in Egypt have documented human and animal exposure to vector-borne bacterial pathogens, but the presence and distribution of these agents in arthropods has not been determined. Between July 2002 and July 2003, fleas were collected from 221 mammals trapped in 17 cities throughout Egypt. A total of 987 fleas were collected, representing four species (Ctenocephalides felis, Echidnophaga gallinacea, Leptopsylla segnis, and Xenopsylla cheopis); 899 of these fleas were X. cheopis from rats (Rattus spp.). Fleas were tested for DNA from Anaplasma spp., Bartonella spp., Coxiella burnetii, Ehrlichia spp., Rickettsia spp., and Yersinia pestis. Rickettsia typhi, the agent of murine typhus, was detected in X. cheopis and L. segnis from rats from nine cities. A spotted-fever group Rickettsia sp. similar to "RF2125" was detected in E. gallinacea, and two unidentified spotted fever group Rickettsia were detected in two X. cheopis. Novel Bartonella genotypes were detected in X. cheopis and L. segnis from three cities. Coxiella burnetii was detected in two fleas. Anaplasma, Ehrlichia, and Y. pestis were not detected.

  9. Molecular characterization of Rickettsia massiliae and Anaplasma platys infecting Rhipicephalus sanguineus ticks and domestic dogs, Buenos Aires (Argentina).

    PubMed

    Cicuttin, Gabriel L; Brambati, Diego F; Rodríguez Eugui, Juan I; Lebrero, Cecilia González; De Salvo, María N; Beltrán, Fernando J; Gury Dohmen, Federico E; Jado, Isabel; Anda, Pedro

    2014-09-01

    Rickettsioses, ehrlichioses and anaplasmoses are emerging diseases that are mainly transmitted by arthropods and that affect humans and animals. The aim of the present study was to use molecular techniques to detect and characterize those pathogens in dogs and ticks from Buenos Aires city. We studied 207 Rhipicephalus sanguineus ticks and 52 canine blood samples from poor neighborhoods of Buenos Aires city. The samples were molecularly screened for the genera Rickettsia, Ehrlichia, and Anaplasma by PCR and sequencing. DNA of Rickettsia massiliae (3.4%) and Anaplasma platys (13.5%) was detected in ticks and blood samples, respectively. For characterization, the positive samples were subjected to amplification of a fragment of the 190-kDa outer membrane protein gene (spotted fever group rickettsiae) and a fragment of the groESL gene (specific for A. platys). A phylogenetic tree was constructed using the neighbor-joining method, revealing that the sequences were closely related to those of strains from other geographic regions. The results indicate that human and animal pathogens are abundant in dogs and their ticks in Buenos Aires city and portray the potentially high risk of human exposure to infection with these agents, especially in poor neighborhoods, where there is close contact with animals in an environment of poor health conditions.

  10. Simultaneous detection of Anaplasma and Ehrlichia species in ruminants and detection of Ehrlichia ruminantium in Amblyomma variegatum ticks by reverse line blot hybridization.

    PubMed

    Bekker, Cornelis P J; de Vos, Sander; Taoufik, Amar; Sparagano, Olivier A E; Jongejan, Frans

    2002-10-22

    The detection of Anaplasma and Ehrlichia species is usually based on species-specific PCR assays, since no assay is yet available which can detect and identify these species simultaneously. To this end, we developed a reverse line blot (RLB) assay for simultaneous detection and identification of Anaplasma and Ehrlichia species in domestic ruminants and ticks. In a PCR the hypervariable V1 region of the 16S ribosomal RNA (rRNA) gene was amplified with a set of primers unique for members of the genera Anaplasma and Ehrlichia [Int. J. Syst. Evol. Microbiol. 51 (2001) 2145]. Amplified PCR products from blood of domestic ruminants or Amblyomma variegatum tick samples were hybridized onto a membrane to which eight species-specific oligonucleotide probes and one Ehrlichia and Anaplasma catch-all oligonucleotide probe were covalently linked. No DNA was amplified from uninfected blood, nor from other hemoparasites such as Theileria annulata, or Babesia bigemina. The species-specific probes did not cross-react with DNA amplified from other species. E. ruminantium, A. ovis and another Ehrlichia were identified by RLB in blood samples collected from small ruminants in Mozambique. Finally, A. variegatum ticks were tested after feeding on E. ruminantium infected sheep. E. ruminantium could be detected in adult ticks even if feeding of nymphs was carried out 3.5 years post-infection. In conclusion, the developed species-specific oligonucleotide probes used in an RLB assay can simultaneously detect and identify several Ehrlichia and Anaplasma species. However, as no quantitative data for the detection limit are available yet, only positive results are interpretable at this stage.

  11. Investigating the Adult Ixodid Tick Populations and Their Associated Anaplasma, Ehrlichia, and Rickettsia Bacteria at a Rocky Mountain Spotted Fever Hotspot in Western Tennessee.

    PubMed

    Trout Fryxell, Rebecca T; Hendricks, Brain M; Pompo, Kimberly; Mays, Sarah E; Paulsen, Dave J; Operario, Darwin J; Houston, Allan E

    2017-08-01

    Ehrlichiosis and rickettsiosis are two common bacterial tick-borne diseases in the southeastern United States. Ehrlichiosis is caused by ehrlichiae transmitted by Amblyomma americanum and rickettsiosis is caused by rickettsiae transmitted by Amblyomma maculatum and Dermacentor variabilis. These ticks are common and have overlapping distributions in the region. The objective of this study was to identify Anaplasma, Ehrlichia, and Rickettsia species associated with questing ticks in a Rocky Mountain spotted fever (RMSF) hotspot, and identify habitats, time periods, and collection methods for collecting questing-infected ticks. Using vegetation drags and CO2-baited traps, ticks were collected six times (May-September 2012) from 100 sites (upland deciduous, bottomland deciduous, grassland, and coniferous habitats) in western Tennessee. Adult collections were screened for Anaplasma and Ehrlichia (simultaneous polymerase chain reaction [PCR]) and Rickettsia using genus-specific PCRs, and resulting positive amplicons were sequenced. Anaplasma and Ehrlichia were only identified within A. americanum (Ehrlichia ewingii, Ehrlichia chaffeensis, Panola Mountain Ehrlichia, and Anaplasma odocoilei sp. nov.); more Ehrlichia-infected A. americanum were collected at the end of June regardless of habitat and collection method. Rickettsia was identified in three tick species; "Candidatus Rickettsia amblyommii" from A. americanum, R. parkeri and R. andeanae from A. maculatum, and R. montanensis ( = montana) from D. variabilis. Overall, significantly more Rickettsia-infected ticks were identified as A. americanum and A. maculatum compared to D. variabilis; more infected-ticks were collected from sites May-July and with dragging. In this study, we report in the Tennessee RMSF hotspot the following: (1) Anaplasma and Ehrlichia are only found in A. americanum, (2) each tick species has its own Rickettsia species, (3) a majority of questing-infected ticks are collected May-July, (4) A

  12. Exploiting social evolution in biofilms.

    PubMed

    Boyle, Kerry E; Heilmann, Silja; van Ditmarsch, Dave; Xavier, Joao B

    2013-04-01

    Bacteria are highly social organisms that communicate via signaling molecules, move collectively over surfaces and make biofilm communities. Nonetheless, our main line of defense against pathogenic bacteria consists of antibiotics-drugs that target individual-level traits of bacterial cells and thus, regrettably, select for resistance against their own action. A possible solution lies in targeting the mechanisms by which bacteria interact with each other within biofilms. The emerging field of microbial social evolution combines molecular microbiology with evolutionary theory to dissect the molecular mechanisms and the evolutionary pressures underpinning bacterial sociality. This exciting new research can ultimately lead to new therapies against biofilm infections that exploit evolutionary cheating or the trade-off between biofilm formation and dispersal.

  13. Energy for lunar resource exploitation

    NASA Astrophysics Data System (ADS)

    Glaser, Peter E.

    1992-02-01

    Humanity stands at the threshold of exploiting the known lunar resources that have opened up with the access to space. America's role in the future exploitation of space, and specifically of lunar resources, may well determine the level of achievement in technology development and global economic competition. Space activities during the coming decades will significantly influence the events on Earth. The 'shifting of history's tectonic plates' is a process that will be hastened by the increasingly insistent demands for higher living standards of the exponentially growing global population. Key to the achievement of a peaceful world in the 21st century, will be the development of a mix of energy resources at a societally acceptable and affordable cost within a realistic planning horizon. This must be the theme for the globally applicable energy sources that are compatible with the Earth's ecology. It is in this context that lunar resources development should be a primary goal for science missions to the Moon, and for establishing an expanding human presence. The economic viability and commercial business potential of mining, extracting, manufacturing, and transporting lunar resource based materials to Earth, Earth orbits, and to undertake macroengineering projects on the Moon remains to be demonstrated. These extensive activities will be supportive of the realization of the potential of space energy sources for use on Earth. These may include generating electricity for use on Earth based on beaming power from Earth orbits and from the Moon to the Earth, and for the production of helium 3 as a fuel for advanced fusion reactors.

  14. Energy for lunar resource exploitation

    NASA Technical Reports Server (NTRS)

    Glaser, Peter E.

    1992-01-01

    Humanity stands at the threshold of exploiting the known lunar resources that have opened up with the access to space. America's role in the future exploitation of space, and specifically of lunar resources, may well determine the level of achievement in technology development and global economic competition. Space activities during the coming decades will significantly influence the events on Earth. The 'shifting of history's tectonic plates' is a process that will be hastened by the increasingly insistent demands for higher living standards of the exponentially growing global population. Key to the achievement of a peaceful world in the 21st century, will be the development of a mix of energy resources at a societally acceptable and affordable cost within a realistic planning horizon. This must be the theme for the globally applicable energy sources that are compatible with the Earth's ecology. It is in this context that lunar resources development should be a primary goal for science missions to the Moon, and for establishing an expanding human presence. The economic viability and commercial business potential of mining, extracting, manufacturing, and transporting lunar resource based materials to Earth, Earth orbits, and to undertake macroengineering projects on the Moon remains to be demonstrated. These extensive activities will be supportive of the realization of the potential of space energy sources for use on Earth. These may include generating electricity for use on Earth based on beaming power from Earth orbits and from the Moon to the Earth, and for the production of helium 3 as a fuel for advanced fusion reactors.

  15. Opportunistic exploitation: an overlooked pathway to extinction.

    PubMed

    Branch, Trevor A; Lobo, Aaron S; Purcell, Steven W

    2013-07-01

    How can species be exploited economically to extinction? Past single-species hypotheses examining the economic plausibility of exploiting rare species have argued that the escalating value of rarity allows extinction to be profitable. We describe an alternative pathway toward extinction in multispecies exploitation systems, termed 'opportunistic exploitation'. In this mode, highly valued species that are targeted first by fishing, hunting, and logging become rare, but their populations can decline further through opportunistic exploitation while more common but less desirable species are targeted. Effectively, expanding exploitation to more species subsidizes the eventual extinction of valuable species at low densities. Managers need to recognize conditions that permit opportunistic depletion and pass regulations to protect highly desirable species when exploitation can expand to other species.

  16. Low temperature affects cattle tick reproduction but does not lead to transovarial transmission of Anaplasma marginale.

    PubMed

    Esteves, E; Pohl, P C; Klafke, G M; Reck, J; Fogaça, A C; Martins, J R; Daffre, S

    2015-12-15

    Anaplasma marginale is an obligate intracellular pathogen that infects the erythrocytes of calves, causing bovine anaplasmosis. This rickettsia is biologically transmitted by several species of ticks. In tropical and subtropical regions of the world, Rhipicephalus microplus is the main vector. Due to their mobility and longevity, the adult males play an important role in the transmission of A. marginale to calves. Some studies have demonstrated that A. marginale can be intrastadially and interstadially transmitted in R. microplus, but the transovarial transmission has not been demonstrated so far. In the present study, we investigated the effects of low temperature on both the A. marginale migration from infected females to their offspring and reproductive parameters of the tick R. microplus. The larvae of R. microplus fed on a calf infected with the strain Jaboticabal of A. marginale. At the end of the parasitic phase, fully engorged females were incubated at either 18°C or 28°C for oviposition. Although A. marginale was detected in the salivary glands of the females, demonstrating that the ticks were successfully infected, the presence of rickettsia was not detected in the offspring. However, the preoviposition period of the non-infected females maintained at 18°C was longer than that of those maintained at 28°C. In addition, the average weight of the mass of eggs as well as the egg production efficiency (ratio of the egg mass weight to the female weight) of the females maintained at 18°C were significantly lower than those of the females incubated at 28°C. There was no larval hatching from the eggs maintained exclusively at 18°C, even at 65 days after female detachment. Hatching occurred only when the eggs maintained at 18°C were transferred to 28°C at 20 days after female detachment (18°C/28°C). We also verified a significantly higher larvae conversion efficiency (ratio of the larvae mass weight to the egg mass weight) in the group of females

  17. Anaplasma marginale Actively Modulates Vacuolar Maturation during Intracellular Infection of Its Tick Vector, Dermacentor andersoni

    PubMed Central

    Thompson, Chelsea Wright; Schneider, David A.; Noh, Susan M.

    2016-01-01

    ABSTRACT Tick-borne transmission of bacterial pathogens in the order Rickettsiales is responsible for diverse infectious diseases, many of them severe, in humans and animals. Transmission dynamics differ among these pathogens and are reflected in the pathogen-vector interaction. Anaplasma marginale has been shown to establish and maintain infectivity within Dermacentor spp. for weeks to months while escaping the complex network of vacuolar peptidases that are responsible for digestion of the tick blood meal. How this prolonged maintenance of infectivity in a potentially hostile environment is achieved has been unknown. Using the natural vector Dermacentor andersoni, we demonstrated that A. marginale-infected tick vacuoles (AmVs) concurrently recruit markers of the early endosome (Rab5), recycling endosome (Rab4 and Rab11), and late endosome (Rab7), are maintained near neutral pH, do not fuse with lysosomes, exclude the protease cathepsin L, and engage the endoplasmic reticulum and Golgi apparatus for up to 21 days postinfection. Maintenance of this safe vacuolar niche requires active A. marginale protein synthesis; in its absence, the AmVs mature into acidic, protease-active phagolysosomes. Identification of this bacterially directed modeling of the tick midgut endosome provides a mechanistic basis for examination of the differences in transmission efficiency observed among A. marginale strains and among vector populations. IMPORTANCE Ticks transmit a variety of intracellular bacterial pathogens that cause significant diseases in humans and animals. For successful transmission, these bacterial pathogens must first gain entry into the tick midgut digestive cells, avoid digestion, and establish a replicative niche without harming the tick vector. Little is known about how this replicative niche is established and maintained. Using the ruminant pathogen A. marginale and its natural tick vector, D. andersoni, this study characterized the features of the A. marginale

  18. Epidemiology and evolution of the genetic variability of Anaplasma marginale in South Africa.

    PubMed

    Mutshembele, Awelani M; Cabezas-Cruz, Alejandro; Mtshali, Moses S; Thekisoe, Oriel M M; Galindo, Ruth C; de la Fuente, José

    2014-10-01

    Bovine anaplasmosis caused by infection of cattle with Anaplasma marginale has been considered to be endemic in South Africa, an assumption based primarily on the distribution of the tick vectors of A. marginale and serological studies on the prevalence of anaplasmosis in Limpopo, Free State, and North West. However, molecular evidence of the distribution of anaplasmosis has only been reported in the Free State province. In order to establish effective control measures for anaplasmosis, epidemiological surveys are needed to define the prevalence and distribution of A. marginale in South Africa. In addition, a proposed control strategy for anaplasmosis is the development of an A. marginale major surface protein 1a (MSP1a)-based vaccine. Nevertheless, regional variations of this gene would need to be characterized prior to vaccine development for South Africa. The objectives of the present study were therefore to conduct a national survey of the prevalence of A. marginale in South Africa, followed by an evaluation of the diversity and evolution of msp1a in South African strains of A. marginale. To accomplish these objectives, species-specific PCR was used to test 250 blood samples from cattle collected from all South African provinces (including 26 districts and municipalities), except the Free State province where similar studies were reported previously. The prevalence of A. marginale ranged from 65% to 100%, except in Northern Cape province where A. marginale was not detected. A correlation was found between the prevalence and genetic diversity of A. marginale MSP1a. Additionally, the genetic diversity of the A. marginale MSP1a was found to evolve under negative and positive selection, and 23 new tandem repeats in South Africa were shown to have evolved from the extant tandem repeat 4. Despite the MSP1a genetic variability, some types of tandem repeats were found to be conserved among the A. marginale strains, and low-variable peptides in MSP1a tandem repeats were

  19. Transcriptional pathways associated with the slow growth phenotype of transformed Anaplasma marginale

    PubMed Central

    2013-01-01

    Background The ability to genetically manipulate bacteria has been fundamentally important for both basic biological discovery and translational research to develop new vaccines and antibiotics. Experimental alteration of the genetic content of prokaryotic pathogens has revealed both expected functional relationships and unexpected phenotypic consequences. Slow growth phenotypes have been reported for multiple transformed bacterial species, including extracellular and intracellular pathogens. Understanding the genes and pathways responsible for the slow growth phenotype provides the opportunity to develop attenuated vaccines as well as bacteriostatic antibiotics. Transformed Anaplasma marginale, a rickettsial pathogen, exhibits slow growth in vitro and in vivo as compared to the parent wild type strain, providing the opportunity to identify the underlying genes and pathways associated with this phenotype. Results Whole genome transcriptional profiling allowed for identification of specific genes and pathways altered in transformed A. marginale. Genes found immediately upstream and downstream of the insertion site, including a four gene operon encoding key outer membrane proteins, were not differentially transcribed between wild type and transformed A. marginale. This lack of significant difference in transcription of flanking genes and the large size of the insert relative to the genome were consistent with a trans rather than a cis effect. Transcriptional profiling across the complete genome identified the most differentially transcribed genes, including an iron transporter, an RNA cleaving enzyme and several genes involved in translation. In order to confirm the trend seen in translation-related genes, K-means clustering and Gene Set Enrichment Analysis (GSEA) were applied. These algorithms allowed evaluation of the behavior of genes as groups that share transcriptional status or biological function. Clustering and GSEA confirmed the initial observations and

  20. The Gaia scientific exploitation networks

    NASA Astrophysics Data System (ADS)

    Figueras, F.; Jordi, C.

    2015-05-01

    On July 2014 the Gaia satellite, placed at L2 since January 2014, finished their commissioning phase and started collecting high accurate scientific data. New and more realistic estimations of the astrometric, photometric and spectroscopic accuracy expected after five years mission operation (2014-2019) have been recently published in the Gaia Science Performance Web page. Here we present the coordination efforts and the activities being conducted through the two GREAT (Gaia Research for European Astronomy Training) European Networks, the GREAT-ESF, a programme supported by the European Science Foundation (2010-2015), and the GREAT-ITN network, from the European Union's Seventh Framework Programme (2011-2015). The main research theme of these networks is to unravel the origin and history of our home galaxy. Emphasis is placed on the research projects being conducted by the Spanish Researchers through these networks, well coordinated by the Red Española de Explotación Científica de Gaia (REG network, with more than 140 participants). Members of the REG play an important role on the collection of complementary spectroscopic data from ground based telescopes, on the development of new tools for an optimal scientific exploitation of Gaia data and on the preparation task to create the Gaia archive.

  1. Exploiting dual otoacoustic emission sources

    NASA Astrophysics Data System (ADS)

    Abdala, Carolina; Kalluri, Radha

    2015-12-01

    Two distinct processes generate otoacoustic emissions (OAEs). Reflection-source emissions, here recorded as stimulus frequency OAEs, are optimally informative at low sound levels and are more sensitive to slight hearing loss; they have been linked to cochlear amplifier gain and tuning. Distortion-source emissions are strongest at moderate-high sound levels and persist despite mild hearing loss; they likely originate in the nonlinear process of hair cell transduction. In this preliminary study, we exploit the unique features of each by generating a combined reflection-distortion OAE profile in normal hearing and hearing-impaired ears. Distortion-product (DP) and stimulus-frequency (SF) OAEs were recorded over a broad range of stimulus levels and frequencies. Individual I/O and transfer functions were generated for both emission types in each ear, and OAE peak strength, compression threshold, and rate of compression were calculated. These combined SFOAE and DPOAE features in normal and hearing-impaired ears may provide a potentially informative and novel index of hearing loss. This is an initial step toward utilizing OAE source in characterizing cochlear function and dysfunction.

  2. Design study for asteroidal exploitation

    NASA Astrophysics Data System (ADS)

    Adams, Carl; Blissit, Jim; Jarrett, Dave; Sanner, Rob; Yanagawa, Koji

    1985-08-01

    A systematic approach to asteroidal exploitation for the 1990 to 2010 time frame is presented as an initial step toward expanding the use of space beyond the space station by providing a source of lower cost materials. With only a limited amount of information known about the asteroids, reconnaissance and exploration phases to determine the exact locations and compositions of several earth-approaching asteroids are required. Earth-based telescopes are used to locate and study the asteroids, while unmanned probes will return samples of asteroidal material to earth for analysis. After these phases are completed, the retrieval of a 35,000 metric ton piece of the asteroid Anteros is undertaken. A cargo transporter uses magnetoplasmadynamic (MPD) arcjets outbound and a mass-driver using asteroidal material inbound. A crew ship uses ion engines. Low thrust trajectories are used for both spacecraft. A materials processing facility will manufacture propellant pellets and retrieve non-propellant materials for spacecraft use. The cost is 1/10th that to transport the same materials from earth to high earth orbit. The project will cost 25 percent less if done in conjunction with a lunar and Martian base.

  3. First molecular evidence of Anaplasma ovis and Rickettsia spp. in keds (Diptera: Hippoboscidae) of sheep and wild ruminants.

    PubMed

    Hornok, Sándor; de la Fuente, José; Biró, Nóra; Fernández de Mera, Isabel G; Meli, Marina L; Elek, Vilmos; Gönczi, Eniko; Meili, Theres; Tánczos, Balázs; Farkas, Róbert; Lutz, Hans; Hofmann-Lehmann, Regina

    2011-10-01

    To evaluate the presence of rickettsial agents in hippoboscid flies with molecular methods, 81 sheep keds (Melophagus ovinus) were collected from 23 sheep, 144 deer keds (Lipoptena cervi) were caught in the environment, and a further 463 and 59 individuals of the latter species were obtained from fresh carcasses of 29 red deer and 17 roe deer, respectively. DNA was extracted individually or in pools. Anaplasma ovis was demonstrated in all examined sheep keds, and from one pool of free-living deer keds. Rickettsia helvetica or other, unidentified rickettsiae were also present in one pool of sheep keds, and in four pools of deer keds from both red deer and roe deer. This is the first account of polymerase chain reaction positivity of hippoboscid flies for A. ovis and rickettsiae. These results raise the possibility that-apart from cattle and roe deer as already reported-sheep and red deer might also play a reservoir role in the epidemiology of rickettsioses.

  4. In vitro establishment and propagation of a Brazilian strain of Anaplasma marginale with appendage in IDE8 (Ixodes scapularis) cells

    PubMed Central

    Bastos, Camila V.; Passos, Lygia M. F.; Vasconcelos, Maria Mercês C.; Ribeiro, Múcio F. B.

    2009-01-01

    A Brazilian isolate of Anaplasma marginale with appendage was successfully established and maintained in vitro in a tick cell line (IDE8). Infection was confirmed by optical and transmission electron microscopy. In addition, primers MSP1aNF2 and MSP1aNR2 amplified products from DNA extracted from infected IDE8 cells. Comparisons with partial sequences of the msp1α gene and the complete genome of A. marginale confirmed that the sequences of amplified fragments were from the A. marginale genome. This is the first establishment of a Brazilian A. marginale isolate in tick cells, representing a new system for biological and molecular studies and also a new source of material for diagnosis and development of vaccines. PMID:24031379

  5. Bacterial Profiling Reveals Novel “Ca. Neoehrlichia”, Ehrlichia, and Anaplasma Species in Australian Human-Biting Ticks

    PubMed Central

    Gofton, Alexander W.; Doggett, Stephen; Ratchford, Andrew; Oskam, Charlotte L.; Paparini, Andrea; Ryan, Una; Irwin, Peter

    2015-01-01

    In Australia, a conclusive aetiology of Lyme disease-like illness in human patients remains elusive, despite growing numbers of people presenting with symptoms attributed to tick bites. In the present study, we surveyed the microbial communities harboured by human-biting ticks from across Australia to identify bacteria that may contribute to this syndrome. Universal PCR primers were used to amplify the V1-2 hyper-variable region of bacterial 16S rRNA genes in DNA samples from individual Ixodes holocyclus (n = 279), Amblyomma triguttatum (n = 167), Haemaphysalis bancrofti (n = 7), and H. longicornis (n = 7) ticks. The 16S amplicons were sequenced on the Illumina MiSeq platform and analysed in USEARCH, QIIME, and BLAST to assign genus and species-level taxonomies. Nested PCR and Sanger sequencing were used to confirm the NGS data and further analyse novel findings. All 460 ticks were negative for Borrelia spp. by both NGS and nested PCR analysis. Two novel “Candidatus Neoehrlichia” spp. were identified in 12.9% of I. holocyclus ticks. A novel Anaplasma sp. was identified in 1.8% of A. triguttatum ticks, and a novel Ehrlichia sp. was identified in both A. triguttatum (1.2%) ticks and a single I. holocyclus (0.6%) tick. Further phylogenetic analysis of novel “Ca. Neoehrlichia”, Anaplasma and Ehrlichia based on 1,265 bp 16S rRNA gene sequences suggests that these are new species. Determining whether these newly discovered organisms cause disease in humans and animals, like closely related bacteria do abroad, is of public health importance and requires further investigation. PMID:26709826

  6. Association of Ehrlichia canis, Hemotropic Mycoplasma spp. and Anaplasma platys and severe anemia in dogs in Thailand.

    PubMed

    Kaewmongkol, Gunn; Lukkana, Nicha; Yangtara, Sarawut; Kaewmongkol, Sarawan; Thengchaisri, Naris; Sirinarumitr, Theerapol; Jittapalapong, Sathaporn; Fenwick, Stanley G

    2017-03-01

    Canine tick-borne bacteria; Ehrlichia canis, hemotropic Mycoplasma spp. and Anaplasma spp., are organisms transmitted by Rhipicephalus sanguineus ticks. However, only a few clinical studies evaluating dogs infected with these organisms and anemia condition have been published. In this study, the potential tick-borne bacteria linked to anemia were investigated in eighty-one blood samples selected from anemic dogs using a broad range nested-PCR of the 16S rRNA gene. Positive results were shown in 12/81 blood specimens (14.81%). Nucleotide sequences from the PCR products were analyzed using BLAST and resulted in identification of Ehrlichia canis (8), Candidatus Mycoplasma haematoparvum (1) and Anaplasma platys (3). Two other PCR assays were used to detect and identify the positive results of these pathogens including a specific PCR for Ehrlichia canis (gp36) and a specific nested-PCR for hemoplasma species (16S rRNA) and the phylogenetic analyses of E. canis and canine hemoplasmas were performed using these two loci. These specific PCRs revealed co-infection of E. canis and Mycoplasma haemocanis in two cases. These two male dogs had presented with jaundice, severe hemolytic anemia, severe thrombocytopenia, leukocytosis, mild azotemia and hepatitis. Ehrlichia canis was detected in a significantly greater number of severe anemia cases (PCV<15%) than moderate or mild anemia cases (PCV 16-29%) (P<0.05) and these severe anemia cases were 7-fold more at risk of having E. canis infections (odds ratio: 7.11, p=0.020). However, no statistical differences were detected between E. canis detection and degrees of thrombocytopenia or leukopenia. From the results of this study, we conclude that the severity of anemia is associated with E. canis infections rather than the severity of thrombocytopenia.

  7. Characterization of Anaplasma marginale subsp. centrale Strains by Use of msp1aS Genotyping Reveals a Wildlife Reservoir

    PubMed Central

    Khumalo, Zamantungwa T. H.; Catanese, Helen N.; Liesching, Nicole; Hove, Paidashe; Collins, Nicola E.; Chaisi, Mamohale E.; Gebremedhin, Assefaw H.; Oosthuizen, Marinda C.

    2016-01-01

    Bovine anaplasmosis caused by the intraerythrocytic rickettsial pathogen Anaplasma marginale is endemic in South Africa. Anaplasma marginale subspecies centrale also infects cattle; however, it causes a milder form of anaplasmosis and is used as a live vaccine against A. marginale. There has been less interest in the epidemiology of A. marginale subsp. centrale, and, as a result, there are few reports detecting natural infections of this organism. When detected in cattle, it is often assumed that it is due to vaccination, and in most cases, it is reported as coinfection with A. marginale without characterization of the strain. A total of 380 blood samples from wild ruminant species and cattle collected from biobanks, national parks, and other regions of South Africa were used in duplex real-time PCR assays to simultaneously detect A. marginale and A. marginale subsp. centrale. PCR results indicated high occurrence of A. marginale subsp. centrale infections, ranging from 25 to 100% in national parks. Samples positive for A. marginale subsp. centrale were further characterized using the msp1aS gene, a homolog of msp1α of A. marginale, which contains repeats at the 5′ ends that are useful for genotyping strains. A total of 47 Msp1aS repeats were identified, which corresponded to 32 A. marginale subsp. centrale genotypes detected in cattle, buffalo, and wildebeest. RepeatAnalyzer was used to examine strain diversity. Our results demonstrate a diversity of A. marginale subsp. centrale strains from cattle and wildlife hosts from South Africa and indicate the utility of msp1aS as a genotypic marker for A. marginale subsp. centrale strain diversity. PMID:27440819

  8. Novel genotypes of Anaplasma bovis, "Candidatus Midichloria" sp. and Ignatzschineria sp. in the Rocky Mountain wood tick, Dermacentor andersoni.

    PubMed

    Dergousoff, Shaun J; Chilton, Neil B

    2011-05-12

    Bovine anaplasmosis, caused by Anaplasma marginale, is a vector-borne disease that is enzootic in many parts of the USA. Although Dermacentor andersoni, a major vector of A. marginale, occurs in Canada, the Canadian cattle herds are currently considered free of bovine anaplasmosis. There have been two outbreaks of the disease in the province of Saskatchewan, but these have been linked to the importation of infected animals. However, the distribution of bovine anaplasmosis may alter with range expansion of the vectors. The aim of the present study was to use molecular techniques to determine if Anaplasma were present in D. andersoni at a locality near its northeastern distributional limit in Saskatchewan. Nested PCR analyses of the bacterial 16S rRNA gene were conducted on the total genomic DNA of 105 individual ticks. Single strand conformation polymorphism analysis and DNA sequencing of the 11 PCR-positive amplicons revealed the presence of three species of bacteria, none of which have been previously reported in D. andersoni. Although no ticks were infected with A. marginale, a novel genotype of A. bovis was detected in eight individuals. This discovery represents the first report of A. bovis in Canada. The potential implications of this finding with respect to animal health and anaplasmosis surveillance in Canada are discussed. The other two bacterial species detected were genetically similar to "Candidatus Midichloria mitochondrii" and Ignatzschineria larvae, the latter of which has been associated with human disease in Europe. Further investigations are needed to determine the prevalence, reservoir hosts, and pathogenicity of the Canadian genotype of A. bovis.

  9. The exploitation of Gestalt principles by magicians.

    PubMed

    Barnhart, Anthony S

    2010-01-01

    Magicians exploit a host of psychological principles in deceiving their audiences. Psychologists have recently attempted to pinpoint the most common psychological tendencies exploited by magicians. This paper highlights two co-occurring principles that appear to be the basis for many popular magic tricks: accidental alignment and good continuation.

  10. Transitioning from AOP to IATA - Exploiting mechanistic ...

    EPA Pesticide Factsheets

    Slide presentation at satellite meeting of the QSAR2016 Meeting on How to Transition from AOP to IATA-Exploiting mechanistic insight for practical decision making. . Slide presentation at satellite meeting of the QSAR2016 Meeting on How to Transition from AOP to IATA-Exploiting mechanistic insight for practical decision making. .

  11. A New Measure of Interpersonal Exploitativeness

    PubMed Central

    Brunell, Amy B.; Davis, Mark S.; Schley, Dan R.; Eng, Abbey L.; van Dulmen, Manfred H.M.; Wester, Kelly L.; Flannery, Daniel J.

    2013-01-01

    Measures of exploitativeness evidence problems with validity and reliability. The present set of studies assessed a new measure [the Interpersonal Exploitativeness Scale (IES)] that defines exploitativeness in terms of reciprocity. In Studies 1 and 2, 33 items were administered to participants. Exploratory and Confirmatory Factor Analysis demonstrated that a single factor consisting of six items adequately assess interpersonal exploitativeness. Study 3 results revealed that the IES was positively associated with “normal” narcissism, pathological narcissism, psychological entitlement, and negative reciprocity and negatively correlated with positive reciprocity. In Study 4, participants competed in a commons dilemma. Those who scored higher on the IES were more likely to harvest a greater share of resources over time, even while controlling for other relevant variables, such as entitlement. Together, these studies show the IES to be a valid and reliable measure of interpersonal exploitativeness. The authors discuss the implications of these studies. PMID:23755031

  12. Infections and risk factors for livestock with species of Anaplasma, Babesia and Brucella under semi-nomadic rearing in Karamoja Region, Uganda.

    PubMed

    Lolli, Chiara; Marenzoni, Maria Luisa; Strona, Paolo; Lappo, Pier Giorgio; Etiang, Patrick; Diverio, Silvana

    2016-03-01

    A survey was conducted to estimate the prevalence of Anaplasma, Babesia and Brucella spp. infections in cattle, goats and sheep in the Karamoja Region of Uganda and to identify possible risk factors existing in this semi-nomadic and pastoral area. Low cost laboratory tests were used to diagnose infections (Rose Bengal test for Brucella spp. antibodies and direct microscopic examination for Anaplasma and Babesia spp.). Multivariable logistic regression models were applied to identify possible risk factors linked to gender, animal species, age (only for cattle) and districts. A total of 3935 cattle, 729 goats and 306 sheep of five districts of the Karamoja Region were tested. Seroprevalence for Brucella was 9.2 % (CI, 95 %: 8.4-10), for Anaplasma 19.5 % (CI 95 %: 18.4-20.6) and for Babesia 16 % (CI 95 %: 15-17.1). Significant differences in infections prevalence were observed against risk factors associated with districts and species. Cattle were the species with higher risk of the infections. Female gender was identified as at risk only for Brucella spp. infection. Cattle more than one year old had greater likelihood to be Brucella seropositive. Co-infections of Anaplasma and Babesia spp. were statistically associated, especially in goats and sheep. Further studies to identify risk factors related to host species and geographical districts are needed. The influence on the semi-nomadic agro-pastoral system in Karamoja of animal raids and animal mixing should be further investigated. Findings were important to sensitize Karamojong undertaking measures on infection control, especially on cattle, which are their main source of food.

  13. Molecular identification of Anaplasma marginale and rickettsial endosymbionts in blood-sucking flies (Diptera: Tabanidae, Muscidae) and hard ticks (Acari: Ixodidae).

    PubMed

    Hornok, Sándor; Földvári, Gábor; Elek, Vilmos; Naranjo, Victoria; Farkas, Róbert; de la Fuente, José

    2008-07-04

    In an attempt to identify the main vector and possible transmission routes of Anaplasma spp. in a region of Hungary with high prevalence of ovine and bovine anaplasmosis, DNA was extracted from 316 haematophagous arthropods (individually or in pools), including 4 species of ixodid ticks, 6 species of tabanid flies and hornflies. Midichloria-like organisms were identified with PCR (amplifying a portion of the 16S rRNA gene) and sequencing from Dermacentor marginatus and Ixodes ricinus. Significantly higher 16S positive D. marginatus individuals were collected in March than in April, suggesting earlier questing of ticks that contain rickettsial agents (thus endosymbionts). Midichloria- and Wolbachia-like organisms were also found in randomly caught horse flies (Tabanus bovinus and T. tergestinus) as well as hornflies (Haematobia irritans), respectively, with 97-99% similarity to sequences deposited in the GenBank. Although all ticks were negative in the Anaplasma spp.-specific msp4 PCR, four individuals of T. bovinus collected near to grazing cattle were positive for Anaplasma marginale. The results of the present study provide the first molecular evidence for the potential mechanical vector role of T. bovinus in the transmission of A. marginale, and broaden the range of haematophagous arthropods harbouring Midichloria-like bacteria, for the first time in any Dermacentor or Tabanus species.

  14. Herbivory eliminates fitness costs of mutualism exploiters.

    PubMed

    Simonsen, Anna K; Stinchcombe, John R

    2014-04-01

    A common empirical observation in mutualistic interactions is the persistence of variation in partner quality and, in particular, the persistence of exploitative phenotypes. For mutualisms between hosts and symbionts, most mutualism theory assumes that exploiters always impose fitness costs on their host. We exposed legume hosts to mutualistic (nitrogen-fixing) and exploitative (non-nitrogen-fixing) symbiotic rhizobia in field conditions, and manipulated the presence or absence of insect herbivory to determine if the costly fitness effects of exploitative rhizobia are context-dependent. Exploitative rhizobia predictably reduced host fitness when herbivores were excluded. However, insects caused greater damage on hosts associating with mutualistic rhizobia, as a consequence of feeding preferences related to leaf nitrogen content, resulting in the elimination of fitness costs imposed on hosts by exploitative rhizobia. Our experiment shows that herbivory is potentially an important factor in influencing the evolutionary dynamic between legumes and rhizobia. Partner choice and host sanctioning are theoretically predicted to stabilize mutualisms by reducing the frequency of exploitative symbionts. We argue that herbivore pressure may actually weaken selection on choice and sanction mechanisms, thus providing one explanation of why host-based discrimination mechanisms may not be completely effective in eliminating nonbeneficial partners.

  15. Global Climate Responses to Anthropogenic Groundwater Exploitation

    NASA Astrophysics Data System (ADS)

    Zeng, Y.; Xie, Z.

    2015-12-01

    In this study, a groundwater exploitation scheme is incorporated into the earth system model, Community Earth System Model 1.2.0 (CESM1.2.0), which is called CESM1.2_GW, and the climatic responses to anthropogenic groundwater withdrawal are then investigated on global scale. The scheme models anthropogenic groundwater exploitation and consumption, which are then divided into agricultural irrigation, industrial use and domestic use. A group of 41-year ensemble groundwater exploitation simulations with six different initial conditions, and a group of ensemble control simulations without exploitation are conducted using the developed model CESM1.2_GW with water supplies and demands estimated. The results reveal that the groundwater exploitation and water consumption cause drying effects on soil moisture in deep layers and wetting effects in upper layers, along with a rapidly declining groundwater table in Central US, Haihe River Basin in China and Northern India and Pakistan where groundwater extraction are most severe in the world. The atmosphere also responds to anthropogenic groundwater exploitation. Cooling effects on lower troposphere appear in large areas of North China Plain and of Northern India and Pakistan. Increased precipitation occurs in Haihe River Basin due to increased evapotranspiration from irrigation. Decreased precipitation occurs in Northern India because water vapor here is taken away by monsoon anomalies induced by anthropogenic alteration of groundwater. The local reducing effects of anthropogenic groundwater exploitation on total terrestrial water storage evinces that water resource is unsustainable with the current high exploitation rate. Therefore, a balance between slow groundwater withdrawal and rapid human economic development must be achieved to maintain a sustainable water resource, especially in over-exploitation regions such as Central US, Northern China, India and Pakistan.

  16. Serological detection of antibodies to Anaplasma spp., Borrelia burgdorferi sensu lato and Ehrlichia canis and of Dirofilaria immitis antigen in dogs from Costa Rica.

    PubMed

    Montenegro, Víctor M; Bonilla, Marta C; Kaminsky, Darwin; Romero-Zúñiga, Juan José; Siebert, Susanne; Krämer, Friederike

    2017-03-15

    In a study in Costa Rica 314 serum samples from dogs throughout all seven provinces were tested using a commercial kit for the detection of circulating antibodies against Anaplasma spp., Borrelia burgdorferi sensu lato and Ehrlichia canis, and of circulating antigen of Dirofilaria immitis. A total of 6.4% (20/314) and 38.2% (120/314) were positive for Anaplasma spp. (An) and E. canis (Ec) antibodies. Overall, 8.0% (25/314) were positive for D. immitis (Di) antigen. One single dog reacted positive with B. burgdorferi s.l. (Bb) antigen (0.3%, 1/314). E. canis positive dogs were detected in all provinces (highest percentages in Guanacaste, Puntarenas [both significantly different compared to the overall] and Limón). Guanacaste and Puntarenas also showed the highest prevalences of Anaplasma spp. (both significantly different compared to the overall). The highest prevalence of D. immitis was detected in Puntarenas (significantly different compared to the overall). Double pathogen exposure (Ec plus An; Ec plus Di; Ec plus Bb) were recorded in 8.9% (28/314). Two dogs showed a triple pathogen exposure (0.6%, 2/314; An, Ec and Di). There was a significant difference between male (11.5%, 18/156) and female (4.4%, 7/158) animals for D. immitis positive results. There was also a significant difference between breed and no breed dogs regarding the characteristics of a general positive test, as well as seropositivity to the single pathogens of Anaplasma spp., E. canis and D. immitis. Finally there was a significant difference in the presence of clinical signs again regarding the characteristics of a general positive test, as well as seropositivity to Anaplasma spp., E. canis and D. immitis. Practitioners in Costa Rica should be aware of the canine vector-borne diseases mentioned as dogs are at risk of becoming infected. Concerning the positive B. burgdorferi s.l. dog, an autochthonous occurrence cannot be confirmed due to a history of adoption and an unusual tattoo number

  17. National Center for Missing and Exploited Children

    MedlinePlus

    ... Team HOPE provides peer and emotional support to families. Contact Us Legal Information DONATE Careers Site Index Copyright © 2016 National Center for Missing & Exploited Children. All rights reserved. This Web site ...

  18. Exploitations and their complications: the necessity of identifying the multiple forms of exploitation in pharmaceutical trials.

    PubMed

    Snyder, Jeremy

    2012-06-01

    Human subject trials of pharmaceuticals in low and middle income countries (LMICs) have been associated with the moral wrong of exploitation on two grounds. First, these trials may include a placebo control arm even when proven treatments for a condition are in use in other (usually wealthier) parts of the world. Second, the trial researchers or sponsors may fail to make a successful treatment developed through the trial available to either the trial participants or the host community following the trial. Many commentators have argued that a single form of exploitation takes place during human subject research in LMICs. These commentators do not, however, agree as to what kind of moral wrong exploitation is or when exploitation is morally impermissible. In this paper, I have two primary goals. First, I will argue for a taxonomy of exploitation that identifies three distinct forms of exploitation. While each of these forms of exploitation has its critics, I will argue that they can each be developed into plausible accounts of exploitation tied to different vulnerabilities and different forms of wrongdoing. Second, I will argue that each of these forms of exploitation can coexist in single situations, including human subject trials of pharmaceuticals. This lesson is important, since different forms of exploitation in a single relationship can influence, among other things, whether the relationship is morally permissible. © 2010 Blackwell Publishing Ltd.

  19. Molecular detection and genetic identification of Babesia bigemina, Theileria annulata, Theileria orientalis and Anaplasma marginale in Turkey.

    PubMed

    Zhou, Mo; Cao, Shinuo; Sevinc, Ferda; Sevinc, Mutlu; Ceylan, Onur; Moumouni, Paul Franck Adjou; Jirapattharasate, Charoonluk; Liu, Mingming; Wang, Guanbo; Iguchi, Aiko; Vudriko, Patrick; Suzuki, Hiroshi; Xuan, Xuenan

    2016-02-01

    Babesia spp., Theileria spp. and Anaplasma spp. are significant tick-borne pathogens of livestock globally. In this study, we investigated the presence and distribution of Babesia bigemina, Theileria annulata, Theileria orientalis and Anaplasma marginale in cattle from 6 provinces of Turkey using species-specific PCR assays. The PCR were conducted using the primers based on the B. bigemina rhoptry-associated protein 1a (BbiRAP-1a), T. annulata merozoite surface antigen-1 (Tams-1), T. orientalis major piroplasm surface protein (ToMPSP) and A. marginale major surface protein 4 (AmMSP4) genes, respectively. Fragments of B. bigemina internal transcribed spacer (BbiITS), T. annulata internal transcribed spacer (TaITS), ToMPSP and AmMSP4 genes were sequenced for phylogenetic analysis. PCR results revealed that the overall infections of A. marginale, T. annulata, B. bigemina and T. orientalis were 29.1%, 18.9%, 11.2% and 5.6%, respectively. The co-infection of two or three pathogens was detected in 29/196 (15.1%) of the cattle samples. The results of sequence analysis indicated that BbiRAP-1a, BbiITS, Tams-1, ToMPSP and AmMSP4 were conserved among the Turkish samples, with 99.76%, 99-99.8%, 99.34-99.78%, 96.9-99.61% and 99.42-99.71% sequence identity values, respectively. In contrast, the Turkish TaITS gene sequences were relatively diverse with 92.3-96.63% identity values. B. bigemina isolates from Turkey were found in the same clade as the isolates from other countries in phylogenetic analysis. On the other hand, phylogenetic analysis based on T. annulata ITS sequences revealed significant differences in the genotypes of T. annulata isolates from Turkey. Additionally, the T. orientalis isolates from Turkish samples were classified as MPSP type 3 genotype. This is the first report of type 3 MPSP in Turkey. Moreover, AmMSP4 isolates from Turkey were found in the same clade as the isolates from other countries. This study provides important data for understanding the

  20. Exploitation in International Paid Surrogacy Arrangements

    PubMed Central

    Wilkinson, Stephen

    2015-01-01

    Abstract Many critics have suggested that international paid surrogacy is exploitative. Taking such concerns as its starting point, this article asks: (1) how defensible is the claim that international paid surrogacy is exploitative and what could be done to make it less exploitative? (2) In the light of the answer to (1), how strong is the case for prohibiting it? Exploitation could in principle be dealt with by improving surrogates' pay and conditions. However, doing so may exacerbate problems with consent. Foremost amongst these is the argument that surrogates from economically disadvantaged countries cannot validly consent because their background circumstances are coercive. Several versions of this argument are examined and I conclude that at least one has some merit. The article's overall conclusion is that while ethically there is something to be concerned about, paid surrogacy is in no worse a position than many other exploitative commercial transactions which take place against a backdrop of global inequality and constrained options, such as poorly‐paid and dangerous construction work. Hence, there is little reason to single surrogacy out for special condemnation. On a policy level, the case for prohibiting international commercial surrogacy is weak, despite legitimate concerns about consent and background poverty. PMID:27471338

  1. Exploitation in International Paid Surrogacy Arrangements.

    PubMed

    Wilkinson, Stephen

    2016-05-01

    Many critics have suggested that international paid surrogacy is exploitative. Taking such concerns as its starting point, this article asks: (1) how defensible is the claim that international paid surrogacy is exploitative and what could be done to make it less exploitative? (2) In the light of the answer to (1), how strong is the case for prohibiting it? Exploitation could in principle be dealt with by improving surrogates' pay and conditions. However, doing so may exacerbate problems with consent. Foremost amongst these is the argument that surrogates from economically disadvantaged countries cannot validly consent because their background circumstances are coercive. Several versions of this argument are examined and I conclude that at least one has some merit. The article's overall conclusion is that while ethically there is something to be concerned about, paid surrogacy is in no worse a position than many other exploitative commercial transactions which take place against a backdrop of global inequality and constrained options, such as poorly-paid and dangerous construction work. Hence, there is little reason to single surrogacy out for special condemnation. On a policy level, the case for prohibiting international commercial surrogacy is weak, despite legitimate concerns about consent and background poverty.

  2. Infection of water buffalo in Rio de Janeiro Brazil with Anaplasma marginale strains also reported in cattle.

    PubMed

    Silva, Jenevaldo B; Cabezas-Cruz, Alejandro; Fonseca, Adivaldo H; Barbosa, José D; de la Fuente, José

    2014-10-15

    Anaplasma marginale is the most prevalent pathogen of cattle in tropical and subtropical regions of the world and causes the disease bovine anaplasmosis. The importance of water buffalo in the world economy is increasing. In addition, while water buffalo may serve as a reservoir host for A. marginale, the susceptibility of this host for A. marginale cattle strains in Brazil has not been reported. The major surface protein 1 alpha (msp1α) gene has been shown to be a stable genetic marker for identification of A. marginale strains. Herein, we analyzed blood samples from 200 water buffalo and identified the A. marginale strains in an endemic area of Rio de Janeiro, Brazil, where ticks were present and water buffalo and cattle co-mingled. Ticks that were feeding on the study buffalo were collected and identified. The prevalence of A. marginale in water buffalo in this study was low (10%). Sequence analysis of the msp1α gene demonstrated the presence of 8 different A. marginale strains. Two A. marginale strains in the water buffalo, (α-β-β-β-Γ) and (α-β-β-Γ), were similar to those reported in cattle from nearby regions. The results of this study suggested that water buffalo in this region are naturally infected with the same strains of A. marginale found in cattle. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. The effect of management factors on the seroprevalence of Anaplasma marginale in Bos indicus cattle in the Mexican tropics.

    PubMed

    Rodríguez-Vivas, R I; Mata-Mendez, Y; Pérez-Gutierrez, E; Wagner, G

    2004-02-01

    A cross-sectional study with a two-stage design and proportional distribution was carried out to determine the effect of management factors on the seroprevalence of Anaplasma marginale in Bos indicus cattle in the Mexican tropics. Serum was obtained from 384 cattle aged 1-2 years on 92 farms. The number of samples was proportionally distributed according to the number of farms in eastern Yucatan. Antibody activity against A. marginale was assessed using the card agglutination test. A primary screening using a 2 x K contingency table of the exposed variables with the outcomes was performed. All variables for which p < 0.20 were included in a fixed-effects log regression. The seroprevalence in the cattle was 69.75% (SE +/- 0.02). Sixty-four per cent of the farms had a seroprevalence > or = 75%. The risks related to managemental factors were stocking density ( > or = 1 animal/ha, OR = 10.94), type of acaricide (pyrethroids, OR = 3.8), dipping interval ( > 60 days, OR = 0.13) and type of veterinary instruments used (needles, scalpels, ear tattoos, and dehorners, OR = 0.17).

  4. Anaplasma marginale infection in a Japanese Black cow 13 years after eradication of Rhipicephalus (Boophilus) microplus in Okinawa, Japan.

    PubMed

    Ooshiro, Mamoru; Zakimi, Satoshi; Matsukawa, Yoshimasa; Yafuso, Makoto; Katagiri, Yoshito; Inokuma, Hisashi

    2009-03-23

    In October 2007, a 15-year-old Japanese Black cow on Ishigaki Island, Okinawa, Japan, was diagnosed with Anaplasma marginale infection based on clinical symptoms, blood examination, smear observation, 16S rRNA and groEL gene sequence analysis, and the result of a CF test. The cow was introduced into the farm from mainland Japan as a calf in 1993, one year before the eradication of Rhipicephalus (Boophilus) microplus, the main vector of A. marginale in Okinawa Prefecture. It is possible that the cow was first infected with A. marginale as a calf in Ishigaki Island and had been persistently infected since then. This is the first reported clinical case of A. marginale infection of cattle since the eradication of R. microplus in Okinawa Prefecture. Additional analysis of major surface protein 1alpha amino acid sequences revealed that the A. marginale Okinawa strain presented four new repeat forms which were not seen in other strains. This indicates that the Okinawa strain may be a unique geographical variant of A. marginale.

  5. Antibody against an Anaplasma marginale MSP5 epitope common to tick and erythrocyte stages identifies persistently infected cattle.

    PubMed Central

    Knowles, D; Torioni de Echaide, S; Palmer, G; McGuire, T; Stiller, D; McElwain, T

    1996-01-01

    A protein epitope of major surface protein 5 (MSP5), defined by monoclonal antibody (MAb) ANAF16C1, is conserved among Anaplasma species (E. S. Visser, T. C. McGuire, G. H. Palmer, W. C. Davis, V. Shkap, E. Pipano, and D. P. Knowles, Jr., Infect. Immun. 60:5139-5144, 1992) and is expressed in the salivary glands of infected ticks. A competitive inhibition ELISA (cELISA) for the detection of bovine anti-MSP5 antibodies was developed by using purified recombinant MSP5 fusion protein and MAb ANAF16C1. The specificity of the recombinant-MSP5 cELISA within North America was established by using 261 serum samples from cattle in the regions of Hawaii and Northern Ontario where anaplasmosis is not endemic and from cattle proven by splenectomy or subinoculation of whole blood into susceptible splenectomized recipients to be uninfected. The maximum percent inhibition by these sera was 18%. Sera known to be positive were obtained from 35 cattle either experimentally inoculated with infected erythrocytes or exposed to infected Dermacentor andersoni ticks. Thirty-four of the 35 serum samples inhibited MAb ANAF16C1 binding by > or = 25%. During acute infection, the MSP5 cELISA detected antibodies prior to or concomitantly with the appearance of rickettsiae in erythrocytes. Antibodies were detectable in sera from persistently infected cattle inoculated as long as 6 years previously. PMID:8862589

  6. Prevalence of antibodies to bluetongue virus and Anaplasma marginale in Montana yearling cattle entering Alberta feedlots: Fall 2001.

    PubMed

    Van Donkersgoed, Joyce; Gertonson, Arnold; Bridges, Marc; Raths, Dick; Dargatz, David; Wagner, Bruce; Boughton, Alice; Knoop, Doug; Walton, Thomas E

    2004-06-01

    A serologic survey was conducted in yearling cattle imported into Alberta feedlots from Montana during October 2001 to estimate the prevalence of antibodies to bluetongue virus (BTV) and Anaplasma marginale in Montana yearling cattle. The apparent prevalence of antibodies to BTV when the competitive enzyme-linked immunosorbent assay (cELISA) was used was 0.37% (21/5608). Test positive cELISA samples were also all positive when tested by virus neutralization (VN) and they reacted to 1 or more BTV serotypes, including 2, 10, 11, 13, and 17. The apparent prevalence of antibodies to A. marginale when a recombinant cELISA (rcELISA) was used with a positive cutoff at 30% inhibition was 1.93% (108/5608). When the rcELISA positive cutoff was at 42% inhibition, the apparent prevalence was 0.73% (41/5608). After the reported sensitivity and specificity of the test had been accounted for, the A. marginale antibody results were consistent with a population that was either free of exposure or had a very low prevalence for A. marginale.

  7. Seroprevalence of Anaplasma marginale in 2 Iowa feedlots and its association with morbidity, mortality, production parameters, and carcass traits.

    PubMed

    Coetzee, Johann F; Schmidt, Peggy L; O'Connor, Annette M; Apley, Michael D

    2010-08-01

    A prospective cohort observational study was conducted to investigate the seroprevalence of Anaplasma marginale in Iowa feedlots and its association with morbidity, mortality, and treatment costs. Blood samples were taken from 659 calves from 31 consigners at processing and classified as seropositive to A. marginale using a competitive enzyme-linked immunosorbent assay (cELISA) with a 30% cutoff. Health and production parameters were modeled by A. marginale serostatus with mixed model regression analysis. The apparent prevalence of seropositive cattle was 15.17% (100/659). When the cELISA positive cutoff was at 42% inhibition, the apparent prevalence was 5.00% (33/659). There was no significant association between A. marginale serostatus and production parameters; however, seropositive status had a weak positive association with undifferentiated fever (P = 0.17). Although prevalence of anaplasmosis in Iowa feedlots is higher than reported in Montana-sourced calves arriving in Canadian feedlots, this was not associated with increased production costs.

  8. Conservation of major surface protein 1 genes of Anaplasma marginale during cyclic transmission between ticks and cattle.

    PubMed

    Bowie, Michael V; de la Fuente, Jose; Kocan, Katherine M; Blouin, Edmour F; Barbet, Anthony F

    2002-01-09

    Bovine anaplasmosis is a rickettsial disease of world-wide economic importance caused by Anaplasma marginale. Several major surface proteins with conserved gene sequences have been examined as potential candidates for vaccines and/or diagnostic assays. Major surface protein 1 (MSP1) is composed of polypeptides MSP1a and MSP1b. MSP1a is expressed from the single copy gene msp1 alpha and MSP1b is expressed by members of the msp1 beta multigene family. In order to determine if the msp1 genes are conserved, primers specific for msp1 alpha, msp1 beta(1), and msp1 beta(2) genes were synthesized and used to amplify msp1 sequences of A. marginale from tick cell cultures, from cattle during acute and chronic infections and from salivary glands of Dermacentor variabilis. Protein sequences of MSP1a, MSP1b(1) and MSP1b(2) were conserved during the life cycle of the parasite. No amino acid changes were observed in MSP1a. However, small variations were observed in the MSP1b(1) and MSP1b(2) protein sequences, which could be attributed to recombination, selection for sub-populations of A. marginale in the vertebrate host and/or PCR errors. Several isolate-specific sequences were also observed. Based on the information obtained in this study, the MSP1 protein appears to be fairly well conserved and a potential vaccine candidate.

  9. Restriction of major surface protein 2 (MSP2) variants during tick transmission of the ehrlichia Anaplasma marginale.

    PubMed

    Rurangirwa, F R; Stiller, D; French, D M; Palmer, G H

    1999-03-16

    Anaplasma marginale is an ehrlichial pathogen of cattle that establishes lifelong persistent infection. Persistence is characterized by rickettsemic cycles in which new A. marginale variant types, defined by the sequence of the expressed msp2 transcripts, emerge. The polymorphic msp2 transcripts encode structurally distinct MSP2 proteins and result in an antigenically diverse and continually changing A. marginale population within the blood. In this manuscript, we used sequence analysis of msp2 transcripts to show that a restricted repertoire of variant types, designated SGV1 and SGV2, is expressed within the tick salivary gland. The same SGV1 and SGV2 variant types were expressed in ticks regardless of the variant types expressed in the blood of infected cattle at the time of acquisition feeding by the ticks. Importantly, subsequent tick transmission to susceptible cattle resulted in acute rickettsemia composed of organisms expressing only the same SGV1 and SGV2 variant types. This indicates that the msp2 expressed by organisms within the tick salivary gland predicts the variant type responsible for acute rickettsemia and disease. This restriction of transmitted A. marginale variant types, in contrast to the marked diversity within persistently infected cattle, supports development of MSP2 vaccines to prevent acute rickettsemia in tick-transmitted infections.

  10. Molecular diagnosis of the tick-borne pathogen Anaplasma marginale in cattle blood samples from Nigeria using qPCR.

    PubMed

    Elelu, Nusirat; Ferrolho, Joana; Couto, Joana; Domingos, Ana; Eisler, Mark C

    2016-12-01

    Tick-borne diseases (TBDs) are some of the most important animal health and management problems in Africa, including Nigeria. This study aims to determine the prevalence of an important TBD, anaplasmosis, in a North-central region of Nigeria. Blood samples were collected from cattle and stored on Whatman FTA(®) cards. Information on village, age and sex associated with each cattle was also recorded. The packed red blood cell volume (PCV) for each blood sample was determined. After DNA extraction, pathogen presence was evaluated by TaqMan(®) based qPCR of which 75.9 % of the cattle tested positive for Anaplasma marginale. Statistical analysis revealed that the presence of A. marginale infection differed significantly between cattle age groups. However, there was no significant difference in the prevalence of this pathogen between the sexes or among cattle grouped by PCV level. Finally, using a highly sensitive molecular method our pioneer study contributes to the improvement of the current knowledge regarding tick-borne pathogens that seriously affect animal health in specific areas of Nigeria.

  11. Molecular detection of Dirofilaria immitis, Hepatozoon canis, Babesia spp., Anaplasma platys and Ehrlichia canis in dogs on Costa Rica.

    PubMed

    Wei, Lanjing; Kelly, Patrick; Ackerson, Kate; El-Mahallawy, Heba S; Kaltenboeck, Bernhard; Wang, Chengming

    2014-03-01

    Although vector-borne diseases are important causes of morbidity and mortality in dogs in tropical areas, there is little information on these conditions in Costa Rica. In PCRs of blood from dogs in Costa Rica, we did not detect DNAs of Rickettsia (R.) felis and Coxiella (C.) burnetii but we did find evidence of infection with Dirofilaria (D.) immitis (9/40, 22.5%), Hepatozoon (H.) canis (15/40, 37.5%), Babesia spp. (10/40, 25%; 2 with B. gibsoni and 8 with B. vogeli), Anaplasma (A.) platys (3/40, 7.5%) and Ehrlichia (E.) canis (20/40, 50%). Nine dogs (22.5%) were free of any vector-borne pathogens while 14 (35%) were infected with a single pathogen, 11 (27.5%) with two, 4 (10%) with three, 1 (2.5%) with four, and 1 (2.5%) with five pathogens. Dogs in Costa Rica are commonly infected with vector-borne agents.

  12. Rethinking exploitation: a process-centered account.

    PubMed

    Jansen, Lynn A; Wall, Steven

    2013-12-01

    Exploitation has become an important topic in recent discussions of biomedical and research ethics. This is due in no small measure to the influence of Alan Wertheimer's path-breaking work on the subject. This paper presents some objections to Wertheimer's account of the concept. The objections attempt to show that his account places too much emphasis on outcome-based considerations and too little on process-based considerations. Building on these objections, the paper develops an alternative process-centered account of the concept. This alternative account of exploitation takes as its point of departure the broadly Kantian notion that it is wrong to use another as an instrument for the advancement of one's own ends. It sharpens this slippery notion and adds a number of refinements to it. The paper concludes by arguing that process-centered accounts of exploitation better illuminate the ethical challenges posed by research on human subjects than outcome-centered accounts.

  13. First report of Anaplasma platys infection in red foxes (Vulpes vulpes) and molecular detection of Ehrlichia canis and Leishmania infantum in foxes from Portugal.

    PubMed

    Cardoso, Luís; Gilad, Matan; Cortes, Helder C E; Nachum-Biala, Yaarit; Lopes, Ana Patrícia; Vila-Viçosa, Maria João; Simões, Margarida; Rodrigues, Paula A; Baneth, Gad

    2015-03-23

    The bacteria Anaplasma platys and Ehrlichia canis and the protozoan Leishmania infantum are vector-borne agents that cause canine vector-borne diseases, some of which are zoonotic. The present survey investigated the prevalence of Anaplasma, Ehrlichia and Leishmania in red foxes (Vulpes vulpes) from Portugal by molecular analysis, in order to evaluate the epidemiological role of these canids as reservoirs of infection. Blood and/or bone marrow samples were collected from 78 red foxes obtained in eight districts of northern, central and southern Portugal. Real-time polymerase chain reactions (PCR) amplified a 123 bp fragment of the 16S rRNA gene of Anaplasma spp. and Ehrlichia spp. and a 265 bp fragment of the L. infantum internal transcribed spacer one (ITS1) region of the rRNA operon evaluated by PCR-high resolution melt analysis (PCR-HRM), with sequencing of the DNA products. A phylogenetic analysis was carried out to compare these to other sequences from Anaplasma spp. and Ehrlichia spp. deposited in GenBank. A. platys was detected in 10 (14.5%) and E. canis in two (2.9%) out of 69 foxes; and L. infantum was detected in one (1.3%) of the 78 foxes. The prevalence of A. platys was significantly different from the prevalence of E. canis (p=0.016) and from that of L. infantum (p=0.002). No co-infections were found in any one of the 78 foxes. No statistically significant differences were found between the type of sample (blood and bone marrow), geographic regions (north/centre and south), age (<2 years and ≥2 years) and gender for any one of the agents. This is the first known report of A. platys in red foxes worldwide, as well as the first molecular evidence of E. canis in foxes from Portugal. The moderate prevalence of A. platys suggests that red foxes may play a role in the epidemiology of infection with this bacterium and serve as a reservoir for domestic dogs.

  14. Child Exploitation: Some Pieces of the Puzzle.

    ERIC Educational Resources Information Center

    Rohlader, Dorothy

    The report addresses the status in North Carolina and in the nation of child exploitation. Legislative and judicial backgrounds of child pornography and child prostitution are reviewed, and difficulties in obtaining statistical data are noted. Law enforcement issues in pornography are cited, and suggestions for further legislation regarding child…

  15. Exploiting a natural auxotrophy for genetic selection.

    PubMed

    Ramage, Elizabeth; Gallagher, Larry; Manoil, Colin

    2012-08-01

    We exploited the natural histidine auxotrophy of Francisella species to develop hisD (encodes histidinol dehydrogenase) as a positive selection marker. A shuttle plasmid (pBR103) carrying Escherichia coli hisD and designed for cloning of PCR fragments replicated in both attenuated and highly virulent Francisella strains. During this work, we formulated a simplified defined growth medium for Francisella novicida.

  16. User interface development for semiautomated imagery exploitation

    NASA Astrophysics Data System (ADS)

    O'Connor, R. P.; Bohling, Edward H.

    1991-08-01

    Operational reconnaissance technical organizations are burdened by greatly increasing workloads due to expanding capabilities for collection and delivery of large-volume near-real- time multisensor/multispectral softcopy imagery. Related to the tasking of reconnaissance platforms to provide the imagery are more stringent timelines for exploiting the imagery in response to the rapidly changing threat environment being monitored. The development of a semi-automated softcopy multisensor image exploitation capability is a critical step toward integrating existing advanced image processing techniques in conjunction with appropriate intelligence and cartographic data for next-generation image exploitation systems. This paper discusses the results of a recent effort to develop computer-assisted aids for the image analyst (IA) in order to rapidly and accurately exploit multispectral/multisensor imagery in combination with intelligence support data and cartographic information for the purpose of target detection and identification. A key challenge of the effort was to design and implement an effective human-computer interface that would satisfy any generic IA task and readily accommodate the needs of a broad range of IAs.

  17. Child Exploitation: Some Pieces of the Puzzle.

    ERIC Educational Resources Information Center

    Rohlader, Dorothy

    The report addresses the status in North Carolina and in the nation of child exploitation. Legislative and judicial backgrounds of child pornography and child prostitution are reviewed, and difficulties in obtaining statistical data are noted. Law enforcement issues in pornography are cited, and suggestions for further legislation regarding child…

  18. Geothermal energy exploitation in New Zealand

    SciTech Connect

    Elder, J.W.

    1980-01-01

    The essential factors, human and technical, which control the operation of geothermal systems, particularly those which allow prediction of behavior during and after exploitation, are sketched. The strategy and co-ordination involved in using New Zealand's geothermal resources for power production are considered. The broader aspects of the technical matters involved in the design of the parasitic plant reservoir system are described. (MHR)

  19. Trolling may intensify exploitation in crappie fisheries

    USGS Publications Warehouse

    Meals, K. O.; Dunn, A. W.; Miranda, Leandro E.

    2012-01-01

    In some parts of the USA, anglers targeting crappies Pomoxis spp. are transitioning from mostly stationary angling with a single pole around submerged structures to using multiple poles while drifting with the wind or under power. This shift in fishing methods could result in a change in catch efficiency, possibly increasing exploitation rates to levels that would be of concern to managers. We studied the catch statistics of anglers fishing while trolling with multiple poles (trollers) and those fishing with single poles (polers) in Mississippi reservoirs. Specifically, we tested whether (1) various catch statistics differed between trollers and polers, (2) catch rates of trollers were related to the number of poles fished, and (3) trollers could raise exploitation rates to potentially unsustainable levels. Results showed that participation in the crappie fisheries was about equally split between polers and trollers. In spring, 90% of crappie anglers were polers; in summer, 85% of crappie anglers were trollers. The size of harvested crappies was similar for the two angler groups, but the catch per hour was almost three times higher for trollers than for polers. Catch rates by trollers were directly correlated to the number of poles fished, although the relationship flattened as the number of poles increased. The average harvest rate for one troller fishing with three poles was similar to the harvest rate obtained by one poler. Simulations predicted that at the existing mix of about 50% polers and 50% trollers and with no restrictions on the number of poles used by trollers, exploitation of crappies is about 1.3 times higher than that in a polers-only fishery; under a scenario in which 100% of crappie anglers were trollers, exploitation was forecasted to increase to about 1.7 times the polers-only rate. The efficiency of trolling for crappies should be of concern to fishery managers because crappie fisheries are mostly consumptive and may increase exploitation

  20. Dissemination and Exploitation: Project Goals beyond Science

    NASA Astrophysics Data System (ADS)

    Hamann, Kristin; Reitz, Anja

    2017-04-01

    Dissemination and Exploitation are essential parts of public funded projects. In Horizon 2020 a plan for the exploitation and dissemination of results (PEDR) is a requirement. The plan should contain a clear vision on the objectives of the project in relation to actions for dissemination and potential exploitation of the project results. The actions follow the basic idea to spread the knowledge and results gathered within the project and face the challenge of how to bring the results into potentially relevant policy circle and how they impact the market. The plan follows the purpose to assess the impact of the project and to address various target groups who are interested in the project results. Simply put, dissemination concentrates on the transfer of knowledge and exploitation on the commercialization of the project. Beyond the question of the measurability of project`s impact, strategies within science marketing can serve purposes beyond internal and external communication. Accordingly, project managers are facing the challenge to implement a dissemination and exploitation strategy that ideally supports the identification of all partners with the project and matches the current discourse of the project`s content within the society, politics and economy. A consolidated plan might unite all projects partners under a central idea and supports the identification with the project beyond the individual research questions. Which applications, strategies and methods can be used to bring forward a PEDR that accompanies a project successfully and allows a comprehensive assessment of the project afterwards? Which hurdles might project managers experience in the dissemination process and which tasks should be fulfilled by the project manager?

  1. Intelligence, mapping, and geospatial exploitation system (IMAGES)

    NASA Astrophysics Data System (ADS)

    Moellman, Dennis E.; Cain, Joel M.

    1998-08-01

    This paper provides further detail to one facet of the battlespace visualization concept described in last year's paper Battlespace Situation Awareness for Force XXI. It focuses on the National Imagery and Mapping Agency (NIMA) goal to 'provide customers seamless access to tailorable imagery, imagery intelligence, and geospatial information.' This paper describes Intelligence, Mapping, and Geospatial Exploitation System (IMAGES), an exploitation element capable of CONUS baseplant operations or field deployment to provide NIMA geospatial information collaboratively into a reconnaissance, surveillance, and target acquisition (RSTA) environment through the United States Imagery and Geospatial Information System (USIGS). In a baseplant CONUS setting IMAGES could be used to produce foundation data to support mission planning. In the field it could be directly associated with a tactical sensor receiver or ground station (e.g. UAV or UGV) to provide near real-time and mission specific RSTA to support mission execution. This paper provides IMAGES functional level design; describes the technologies, their interactions and interdependencies; and presents a notional operational scenario to illustrate the system flexibility. Using as a system backbone an intelligent software agent technology, called Open Agent ArchitectureTM (OAATM), IMAGES combines multimodal data entry, natural language understanding, and perceptual and evidential reasoning for system management. Configured to be DII COE compliant, it would utilize, to the extent possible, COTS applications software for data management, processing, fusion, exploitation, and reporting. It would also be modular, scaleable, and reconfigurable. This paper describes how the OAATM achieves data synchronization and enables the necessary level of information to be rapidly available to various command echelons for making informed decisions. The reasoning component will provide for the best information to be developed in the timeline

  2. Identification of a cell epitope that is globally conserved among outer membrane proteins (OMPs) OMP7, OMP8, and OMP9 of anaplasma marginale strains and with OMP7 from the A. marginale subsp. centrale vaccine strain

    USDA-ARS?s Scientific Manuscript database

    Within the protective outer membrane fraction of Anaplasma marginale, several vaccine candidates have emerged, including a family of outer membrane proteins (OMPs) 7-9, which share sequence identity with each other and with the single protein OMP7 in the vaccine strain A. marginale subsp. centrale. ...

  3. Loss of immunization-induced epitope-specific CD4 T-cell response following anaplasma marginale infection requires presence of the T-cell epitope on the pathogen and is not associated with an increase in lymphocytes

    USDA-ARS?s Scientific Manuscript database

    We have shown that in cattle previously immunized with outer membrane proteins, infection with Anaplasma marginale induces a functionally exhausted CD4 T-cell response to the A. marginale immunogen. Furthermore, T-cell responses following infection in nonimmunized cattle had a delayed onset and were...

  4. Cooperation of PD-1 and LAG-3 Contributes to T-Cell Exhaustion in Anaplasma marginale-Infected Cattle

    PubMed Central

    Okagawa, Tomohiro; Konnai, Satoru; Deringer, James R.; Ueti, Massaro W.; Scoles, Glen A.; Murata, Shiro; Ohashi, Kazuhiko

    2016-01-01

    The CD4+ T-cell response is central for the control of Anaplasma marginale infection in cattle. However, the infection induces a functional exhaustion of antigen-specific CD4+ T cells in cattle immunized with A. marginale outer membrane proteins or purified outer membranes (OMs), which presumably facilitates the persistence of this rickettsia. In the present study, we hypothesize that T-cell exhaustion following infection is induced by the upregulation of immunoinhibitory receptors on T cells, such as programmed death 1 (PD-1) and lymphocyte activation gene 3 (LAG-3). OM-specific T-cell responses and the kinetics of PD-1-positive (PD-1+) LAG-3+ exhausted T cells were monitored in A. marginale-challenged cattle previously immunized with OMs. Consistent with data from previous studies, OM-specific proliferation of peripheral blood mononuclear cells (PBMCs) and interferon gamma (IFN-γ) production were significantly suppressed in challenged animals by 5 weeks postinfection (wpi). In addition, bacteremia and anemia also peaked in these animals at 5 wpi. Flow cytometric analysis revealed that the percentage of PD-1+ LAG-3+ T cells in the CD4+, CD8+, and γδ T-cell populations gradually increased and also peaked at 5 wpi. A large increase in the percentage of LAG-3+ γδ T cells was also observed. Importantly, in vitro, the combined blockade of the PD-1 and LAG-3 pathways partially restored OM-specific PBMC proliferation and IFN-γ production at 5 wpi. Taken together, these results indicate that coexpression of PD-1 and LAG-3 on T cells contributes to the rapid exhaustion of A. marginale-specific T cells following infection and that these immunoinhibitory receptors regulate T-cell responses during bovine anaplasmosis. PMID:27430272

  5. Low genetic diversity associated with low prevalence of Anaplasma marginale in water buffaloes in Marajó Island, Brazil.

    PubMed

    Silva, Jenevaldo B; Fonseca, Adivaldo H; Barbosa, José D; Cabezas-Cruz, Alejandro; de la Fuente, José

    2014-10-01

    The rickettsia Anaplasma marginale is the etiologic agent of bovine anaplasmosis, an important tick-borne disease affecting cattle in tropical and subtropical regions of the world. In endemic regions, the genetic diversity of this pathogen is usually related to the high prevalence of the disease in cattle. The major surface protein 1 alpha (MSP1a) has been used as a marker to characterize the genetic diversity and for geographical identification of A. marginale strains. The present study reports the characterization of A. marginale MSP1a diversity in water buffaloes. Blood samples were collected from 200 water buffaloes on Marajó Island, Brazil where the largest buffalo herd is located in the Western hemisphere. Fifteen buffaloes (7.5%) were positive for A. marginale msp1α by PCR. Four different strains of A. marginale with MSP1a tandem repeat structures (4-63-27), (162-63-27), (78-24-24-25-31) and (τ-10-10-15) were found, being (4-63-27) the most common. MSP1a tandem repeats composition in buffalos and phylogenetic analysis using msp1α gene showed that the A. marginale strains identified in buffaloes are closely related to A. marginale strains from cattle. The results demonstrated low genetic diversity of A. marginale associated with low bacterial prevalence in buffaloes and suggested that buffaloes may be reservoirs of this pathogen for cattle living in the same area. The results also suggested that mechanical transmission and not biological transmission by ticks might be playing the major role for pathogen circulation among water buffaloes in Marajó Island, Brazil. Copyright © 2014 Elsevier GmbH. All rights reserved.

  6. Cattle experimentally infected by Anaplasma marginale: Influence of splenectomy on disease pathogenesis, oxidative profile, and antioxidant status.

    PubMed

    Doyle, Rovaina L; França, Raqueli T; Oliveira, Camila B; Rezer, João F P; Klafke, Guilherme M; Martins, João R; Santos, Andrea P; do Nascimento, Naíla C; Mesick, Joanne B; Lopes, Sonia T A; Leal, Daniela B R; Da Silva, Aleksandro S; Andrade, Cinthia M

    2016-06-01

    Bovine anaplasmosis is caused by the obligate intraerythrocytic bacteria Anaplasma marginale. These bacteria are transmitted by tick species such as Rhipicephalus (Boophilus) microplus, blood-sucking insects, and fomites (needles, clippers, and other blood contaminated equipment). During the acute phase of infection, animals may develop fever, anemia, jaundice, and hepatosplenomegaly. The aims of this study are to quantify the bacteremia by quantitative PCR in eight naïve calves experimentally infected by A. marginale [splenectomized (n = 4), and intact/non-splenectomized (n = 4)], and to correlate these findings with markers of oxidative stress on days 0, 8, 15, 21 and 23 post-infection. Complete blood counts (CBC) were performed in both groups. Lipid peroxidation was estimated by quantifying thiobarbituric acid reactive substances (TBARS); and non-enzymatic antioxidants were assessed by erythrocyte content of non-protein thiols (NPSH). There were no significant differences in complete blood counts (CBC) between the two groups. However, both groups had a slight decrease on packet cell volume (PCV), erythrocytes and hemoglobin concentration, as well as an increase in total leukocyte counts due to elevated lymphocytes when comparing pre and post-infection with A. marginale. Progressive increase on TBARS levels and concomitant decrease on NPSH content were observed in all animals, without significant differences between splenectomized and intact animals. A positive correlation between bacteremia and TBARS, and a negative correlation between bacteremia and NPSH were observed in both groups with higher correlation for NPSH in splenectomized animals. A negative correlation between TBARS and NPSH levels was observed in both groups indicating lipid peroxidation without a non-enzymatic antioxidant response. The results of experimental infection by A. marginale in cattle showed that bacteremia has an impact on lipid peroxidation regardless of the splenectomy.

  7. Molecular detection of Anaplasma platys infection in free-roaming dogs and ticks from Kenya and Ivory Coast.

    PubMed

    Matei, Ioana Adriana; D'Amico, Gianluca; Yao, Patrick K; Ionică, Angela Monica; Kanyari, Paul W N; Daskalaki, Aikaterini Alexandra; Dumitrache, Mirabela Oana; Sándor, Attila D; Gherman, Călin Mircea; Qablan, Moneeb; Modrý, David; Mihalca, Andrei Daniel

    2016-03-16

    Anaplasma platys is a bacterium parasitic in the canine platelets, representing the causative agent of canine cyclic thrombocytopenia, with a worldwide distribution, mainly in tropical countries. The agent has zoonotic potential, being reported in several human clinical cases. The suspected vector is the tick Rhipicephalus sanguineus (sensu lato), widely distributed in animals and humans in the tropical regions of South America, Africa, Asia and Australia, but also in southern Europe. Only few scattered data concerning the epidemiology of A. platys in sub-Saharan Africa are available. The aim of our study was to evaluate the epidemiological status of A. platys in dogs and cats from rural communities in eastern and western Africa, where dogs and their ticks live in close contact with humans. A total of 216 canine and 22 feline blood samples and ticks feeding on them were collected in 2013 and 2014 from eight localities in Ivory Coast and four localities in Kenya. PCR was performed using specific primers targeting a portion of the 16S rRNA gene, followed by sequencing. The main results highlight the high prevalence of A. platys infection in dogs in both Eastern and Western Africa and report it for the first time in Eastern Africa and in Rhipicephalus camicasi. The presence of A. platys DNA in R. camicasi feeding on negative hosts together with the relatively high prevalence of A. platys, correlated with the absence of the probable vector Rhipicephalus sanguineus (s.l.) in Kenyan Island suggest the involvement of other tick species in the transmission of A. platys.

  8. Development of a multilocus sequence typing scheme for the study of Anaplasma marginale population structure over space and time.

    PubMed

    Guillemi, Eliana C; Ruybal, Paula; Lia, Verónica; Gonzalez, Sergio; Lew, Sergio; Zimmer, Patricia; Lopez Arias, Ludmila; Rodriguez, Jose L; Rodriguez, Sonia Y; Frutos, Roger; Wilkowsky, Silvina E; Farber, Marisa D

    2015-03-01

    Bovine Anaplasmosis caused by Anaplasma marginale is a worldwide disease prevalent in tropical and subtropical regions where Rhipicephalus microplus is considered the most significant biological vector. Molecular markers previously applied for A. marginale typing are efficient for isolate discrimination but they are not a suitable tool for studying population structure and dynamics. Here we report the development of an MLST scheme based on the study of seven genes: dnaA, ftsZ, groEl, lipA, recA, secY and sucB. Five annotated genomes (Saint Maries, Florida, Mississippi, Puerto Rico and Virginia) and 53 bovine blood samples from different world regions were analyzed. High nucleotide diversity and a large proportion of synonymous substitutions, indicative of negative selection resulted from DnaSP 5.00.02 package application. Recombination events were detected in almost all genes, this evidence together with the coexistence of more than one A. marginale strain in the same sample might suggest the superinfection phenomena as a potential source of variation. The allelic profile analysis performed through GoeBURST shown two main CC that did not support geography. In addition, the AMOVA test confirmed the occurrence of at least two main genetically divergent groups. The composition of the emergent groups reflected the impact of both historical and environmental traits on A. marginale population structure. Finally, a web-based platform "Galaxy MLST-Pipeline" was developed to automate DNA sequence editing and data analysis that together with the Data Base are freely available to users. The A. marginale MLST scheme developed here is a valuable tool with a high discrimination power, besides PCR based strategies are still the better choice for epidemiological intracellular pathogens studies. Finally, the allelic profile describe herein would contribute to uncover the mechanisms in how intracellular pathogens challenge virulence paradigm.

  9. Detection of Anaplasma platys in dogs and Rhipicephalus sanguineus group ticks by a quantitative real-time PCR.

    PubMed

    Ramos, Rafael Antonio Nascimento; Latrofa, Maria Stefania; Giannelli, Alessio; Lacasella, Vita; Campbell, Bronwyn Evelyn; Dantas-Torres, Filipe; Otranto, Domenico

    2014-09-15

    Anaplasma platys is an obligate intracellular, tick-borne pathogen of dogs, which causes canine infectious cyclic thrombocytopenia (CICT). The vector role of Rhipicephalus sanguineus group ticks has been only suggested, but definitive evidence is lacking. This study aimed to detect and quantify A. platys DNA in infected dogs and in their respective ticks through a quantitative real-time PCR assay. From March to May 2009, blood and tick samples from dogs residing in a CICT-endemic area were collected and molecularly analysed. Differences (p<0.05) were detected in the bacterial load between tick nymphs (3.5 × 10(-2)± 2.5 × 10(-2)) and adults (female: 1.2 × 10(-1) ± 1.1 × 10(-1); male: 9.3 × 10(-2)± 1.2 × 10(-2)) and between unengorged (1.1 × 10(-1) ± 2.8 × 10(-2)) and partially or fully engorged ticks (1.9 × 10(-1) ± 2.7 × 10(-2)). No difference was found between ticks collected from positive (1.1 × 10(-1)± 0.6 × 10(-1)) and negative (1.2 × 10(-1) ± 0.3 × 10(-1)) dogs (p>0.05). The mean bacterial load detected in positive dogs was lower than that in their respective ticks (p>0.05). This study provides circumstantial evidence of the putative role of Rhipicephalus sp. I as a vector of this pathogen.

  10. Exploiting host immunity: the Salmonella paradigm

    PubMed Central

    Behnsen, Judith; Perez-Lopez, Araceli; Nuccio, Sean-Paul; Raffatellu, Manuela

    2014-01-01

    Pathogens have evolved clever strategies to evade and in some cases exploit the attacks of an activated immune system. Salmonella enterica is one such pathogen, exploiting multiple aspects of host defense to promote its replication in the host. Here we review recent findings on the mechanisms by which Salmonella establishes systemic and chronic infection, including strategies involving manipulation of innate immune signaling and inflammatory forms of cell death, as well as immune evasion by establishing residency in M2 macrophages. We also examine recent evidence showing that the oxidative environment and the high levels of antimicrobial proteins produced in response to localized Salmonella gastrointestinal infection enable the pathogen to successfully outcompete the resident gut microbiota. PMID:25582038

  11. Exploiting Non-Markovianity for Quantum Control

    NASA Astrophysics Data System (ADS)

    Reich, Daniel M.; Katz, Nadav; Koch, Christiane P.

    2015-07-01

    Quantum technology, exploiting entanglement and the wave nature of matter, relies on the ability to accurately control quantum systems. Quantum control is often compromised by the interaction of the system with its environment since this causes loss of amplitude and phase. However, when the dynamics of the open quantum system is non-Markovian, amplitude and phase flow not only from the system into the environment but also back. Interaction with the environment is then not necessarily detrimental. We show that the back-flow of amplitude and phase can be exploited to carry out quantum control tasks that could not be realized if the system was isolated. The control is facilitated by a few strongly coupled, sufficiently isolated environmental modes. Our paradigmatic example considers a weakly anharmonic ladder with resonant amplitude control only, restricting realizable operations to SO(N). The coupling to the environment, when harnessed with optimization techniques, allows for full SU(N) controllability.

  12. Exploiting Quantum Resonance to Solve Combinatorial Problems

    NASA Technical Reports Server (NTRS)

    Zak, Michail; Fijany, Amir

    2006-01-01

    Quantum resonance would be exploited in a proposed quantum-computing approach to the solution of combinatorial optimization problems. In quantum computing in general, one takes advantage of the fact that an algorithm cannot be decoupled from the physical effects available to implement it. Prior approaches to quantum computing have involved exploitation of only a subset of known quantum physical effects, notably including parallelism and entanglement, but not including resonance. In the proposed approach, one would utilize the combinatorial properties of tensor-product decomposability of unitary evolution of many-particle quantum systems for physically simulating solutions to NP-complete problems (a class of problems that are intractable with respect to classical methods of computation). In this approach, reinforcement and selection of a desired solution would be executed by means of quantum resonance. Classes of NP-complete problems that are important in practice and could be solved by the proposed approach include planning, scheduling, search, and optimal design.

  13. Tribal children are most exploited - UNICEF.

    PubMed

    A workshop sponsored by the UN Children's Fund in the Philippines examined the status of the children of indigenous people and found that exploitation of the assets of indigenous people in the name of development has resulted in social inequalities that have damaged the indigenous children. As examples of the disregard for the human rights of the children, participants cited projects in Davao, Boracay, and Benguet that have displaced native children. These include mining schemes that have "raped" ancestral lands, large-scale agricultural enterprises, promotion of tourism, and creation of hydroelectric dams. The children rarely benefit at all from any of these projects as their families are moved from a position of isolated independence to one of exploited dependence. Social changes accompanying development ruin traditional culture without providing a better or even similar basis of existence.

  14. Molecular identification of Anaplasma marginale in two autochthonous South American wild species revealed an identical new genotype and its phylogenetic relationship with those of bovines.

    PubMed

    Guillemi, Eliana C; de la Fourniere, Sofía; Orozco, Marcela; Peña Martinez, Jorge; Correa, Elena; Fernandez, Javier; Lopez Arias, Ludmila; Paoletta, Martina; Corona, Belkis; Pinarello, Valérie; Wilkowsky, Silvina E; Farber, Marisa D

    2016-05-26

    Anaplasma marginale is a well-known cattle pathogen of tropical and subtropical world regions. Even though, this obligate intracellular bacterium has been reported in other host species different than bovine, it has never been documented in Myrmecophaga tridactyla (giant anteater) or Hippocamelus antisense (taruca), which are two native endangered species. Samples from two sick wild animals: a Myrmecophaga tridactyla (blood) and a Hippocamelus antisense (blood and serum) were studied for the presence of A. marginale DNA through msp5 gene fragment amplification. Further characterization was done through MSP1a tandem repeats analysis and MLST scheme and the genetic relationship among previously characterized A. marginale sequences were studied by applying, eBURST algorithm and AMOVA analysis. Anaplasma marginale DNA was identified in the Myrmecophaga tridactyla and Hippocamelus antisense samples. Through molecular markers, we identified an identical genotype in both animals that was not previously reported in bovine host. The analysis through eBURST and AMOVA revealed no differentiation between the taruca/anteater isolate and the bovine group. In the present publication we report the identification of A. marginale DNA in a novel ruminant (Hippocamelus antisense) and non-ruminant (Myrmecophaga tridactyla) host species. Genotyping analysis of isolates demonstrated the close relatedness of the new isolate with the circulation population of A. marginale in livestock. Further analysis is needed to understand whether these two hosts contribute to the anaplasmosis epidemiology.

  15. Exploiting Virtual Synchrony in Distributed Systems

    DTIC Science & Technology

    1987-02-01

    state, by having each current member reply with a data structure encoding this information. In a virtually syn- chronous sense , the new member joins...Exlitn Virua Sychon inf :en.et P. Birma ThmbA.Jsp TECHNICAL REPORTF Department of Computer Science Cornell University Ithaca, New York Exploiting...1 Department of Computer Science Justifi: i ____ Cornell University Ithaca, New York 14853-7501 Distrit ui. _w/ - Avail * t ] ’ - Dist S;. This work

  16. Macropinocytosis Exploitation by Cancers and Cancer Therapeutics

    PubMed Central

    Ha, Kevin D.; Bidlingmaier, Scott M.; Liu, Bin

    2016-01-01

    Macropinocytosis has long been known as a primary method for cellular intake of fluid-phase and membrane-bound bulk cargo. This review seeks to re-examine the latest studies to emphasize how cancers exploit macropinocytosis to further their tumorigenesis, including details in how macropinocytosis can be adapted to serve diverse functions. Furthermore, this review will also cover the latest endeavors in targeting macropinocytosis as an avenue for novel therapeutics. PMID:27672367

  17. DANDRUFF: THE MOST COMMERCIALLY EXPLOITED SKIN DISEASE

    PubMed Central

    Ranganathan, S; Mukhopadhyay, T

    2010-01-01

    The article discuss in detail about the prevalence, pathophysiology, clinical manifestations of dandruff including the etio-pathology. The article also discusses in detail about various treatment methods available for dandruff. The status of dandruff being amphibious – a disease/disorder, and relatively less medical intervention is sought after for the treatment, dandruff is the most commercially exploited skin and scalp disorder/disease by personal care industries. PMID:20606879

  18. SAIM: a mobile multisensor image exploitation system

    NASA Astrophysics Data System (ADS)

    Devambez, Francois

    2000-11-01

    The control of information is an essential part of operations. Technology allows today a near real time surveillance capacity, over wide areas, due to sensor performances, communication networks. The system presented herein has been developed by Thomson-Csf, under contract with the French MOD to give to the decision makers the right information, in a very short delay, and prepare support information, to help for decision. The SAIM, Mobile Multisensor Image Exploitation Ground System, uses near real time acquisition units, very large data base management, data processing, including fusion and decision aiding tools, and communication networks. It then helps for all the steps of exploitation of data incoming from image sensors, form preparation of the reconnaissance mission to the dissemination of intelligence. The SAIM system is in operations in the French Air Force, and soon in the French Navy and the French Army. Initially defined for the specific use of French Recce sensors, the SAIM is now intended to be widely used for the exploitation of UAV and battle field MTI and SAR surveillance systems.

  19. Exploitation of subsea gas hydrate reservoirs

    NASA Astrophysics Data System (ADS)

    Janicki, Georg; Schlüter, Stefan; Hennig, Torsten; Deerberg, Görge

    2016-04-01

    Natural gas hydrates are considered to be a potential energy resource in the future. They occur in permafrost areas as well as in subsea sediments and are stable at high pressure and low temperature conditions. According to estimations the amount of carbon bonded in natural gas hydrates worldwide is two times larger than in all known conventional fossil fuels. Besides technical challenges that have to be overcome climate and safety issues have to be considered before a commercial exploitation of such unconventional reservoirs. The potential of producing natural gas from subsea gas hydrate deposits by various means (e.g. depressurization and/or injection of carbon dioxide) is numerically studied in the frame of the German research project »SUGAR«. The basic mechanisms of gas hydrate formation/dissociation and heat and mass transport in porous media are considered and implemented into a numerical model. The physics of the process leads to strong non-linear couplings between hydraulic fluid flow, hydrate dissociation and formation, hydraulic properties of the sediment, partial pressures and seawater solution of components and the thermal budget of the system described by the heat equation. This paper is intended to provide an overview of the recent development regarding the production of natural gas from subsea gas hydrate reservoirs. It aims at giving a broad insight into natural gas hydrates and covering relevant aspects of the exploitation process. It is focused on the thermodynamic principles and technological approaches for the exploitation. The effects occurring during natural gas production within hydrate filled sediment layers are identified and discussed by means of numerical simulation results. The behaviour of relevant process parameters such as pressure, temperature and phase saturations is described and compared for different strategies. The simulations are complemented by calculations for different safety relevant problems.

  20. Gaze interaction in UAS video exploitation

    NASA Astrophysics Data System (ADS)

    Hild, Jutta; Brüstle, Stefan; Heinze, Norbert; Peinsipp-Byma, Elisabeth

    2013-05-01

    A frequently occurring interaction task in UAS video exploitation is the marking or selection of objects of interest in the video. If an object of interest is visually detected by the image analyst, its selection/marking for further exploitation, documentation and communication with the team is a necessary task. Today object selection is usually performed by mouse interaction. As due to sensor motion all objects in the video move, object selection can be rather challenging, especially if strong and fast and ego-motions are present, e.g., with small airborne sensor platforms. In addition to that, objects of interest are sometimes too shortly visible to be selected by the analyst using mouse interaction. To address this issue we propose an eye tracker as input device for object selection. As the eye tracker continuously provides the gaze position of the analyst on the monitor, it is intuitive to use the gaze position for pointing at an object. The selection is then actuated by pressing a button. We integrated this gaze-based "gaze + key press" object selection into Fraunhofer IOSB's exploitation station ABUL using a Tobii X60 eye tracker and a standard keyboard for the button press. Representing the object selections in a spatial relational database, ABUL enables the image analyst to efficiently query the video data in a post processing step for selected objects of interest with respect to their geographical and other properties. An experimental evaluation is presented, comparing gaze-based interaction with mouse interaction in the context of object selection in UAS videos.

  1. Algorithms exploiting ultrasonic sensors for subject classification

    NASA Astrophysics Data System (ADS)

    Desai, Sachi; Quoraishee, Shafik

    2009-09-01

    Proposed here is a series of techniques exploiting micro-Doppler ultrasonic sensors capable of characterizing various detected mammalian targets based on their physiological movements captured a series of robust features. Employed is a combination of unique and conventional digital signal processing techniques arranged in such a manner they become capable of classifying a series of walkers. These processes for feature extraction develops a robust feature space capable of providing discrimination of various movements generated from bipeds and quadrupeds and further subdivided into large or small. These movements can be exploited to provide specific information of a given signature dividing it in a series of subset signatures exploiting wavelets to generate start/stop times. After viewing a series spectrograms of the signature we are able to see distinct differences and utilizing kurtosis, we generate an envelope detector capable of isolating each of the corresponding step cycles generated during a walk. The walk cycle is defined as one complete sequence of walking/running from the foot pushing off the ground and concluding when returning to the ground. This time information segments the events that are readily seen in the spectrogram but obstructed in the temporal domain into individual walk sequences. This walking sequence is then subsequently translated into a three dimensional waterfall plot defining the expected energy value associated with the motion at particular instance of time and frequency. The value is capable of being repeatable for each particular class and employable to discriminate the events. Highly reliable classification is realized exploiting a classifier trained on a candidate sample space derived from the associated gyrations created by motion from actors of interest. The classifier developed herein provides a capability to classify events as an adult humans, children humans, horses, and dogs at potentially high rates based on the tested sample

  2. Art Therapy Exhibitions: Exploitation or Advocacy?

    PubMed

    Davis, Terri

    2017-01-01

    Promoting awareness of human trafficking by sharing trauma survivors' art and summaries of their life stories suggests ethical complexities that have been typically neglected by bioethicists. Although these survivors voluntarily share the objects they created during art therapy sessions, they are still at risk of harm, including further exploitation, due to their vulnerability, high rates of victim sensitivity, and the mental health consequences of their traumatic experiences. While some argue that the benefits of sublimation and art therapy for human trafficking survivors make sharing their art worth the risk, anti-trafficking organizations and supporters of such art exhibitions have responsibilities to be trauma informed. © 2017 American Medical Association. All Rights Reserved.

  3. Competing Discourses about Youth Sexual Exploitation in Canadian News Media.

    PubMed

    Saewyc, Elizabeth M; Miller, Bonnie B; Rivers, Robert; Matthews, Jennifer; Hilario, Carla; Hirakata, Pam

    2013-10-01

    Media holds the power to create, maintain, or break down stigmatizing attitudes, which affect policies, funding, and services. To understand how Canadian news media depicts the commercial sexual exploitation of children and youth, we examined 835 Canadian newspaper articles from 1989-2008 using a mixed methods critical discourse analysis approach, comparing representations to existing research about sexually exploited youth. Despite research evidence that equal rates of boys and girls experience exploitation, Canadian news media depicted exploited youth predominantly as heterosexual girls, and described them alternately as victims or workers in a trade, often both in the same story. News media mentioned exploiters far less often than victims, and portrayed them almost exclusively as male, most often called 'customers' or 'consumers,' and occasionally 'predators'; in contrast, research has documented the majority of sexually exploited boys report female exploiters. Few news stories over the past two decades portrayed the diversity of victims, perpetrators, and venues of exploitation reported in research. The focus on victims but not exploiters helps perpetuate stereotypes of sexual exploitation as business or a 'victimless crime,' maintains the status quo, and blurs responsibility for protecting youth under the UN Convention on the Rights of the Child. Health care providers and researchers can be advocates for accuracy in media coverage about sexual exploitation; news reporters and editors should focus on exploiters more than victims, draw on existing research evidence to avoid perpetuating stereotypes, and use accurate terms, such as commercial sexual exploitation, rather than terms related to business or trade.

  4. Molecular detection and genetic diversity of bovine Babesia spp., Theileria orientalis, and Anaplasma marginale in beef cattle in Thailand.

    PubMed

    Jirapattharasate, Charoonluk; Adjou Moumouni, Paul Franck; Cao, Shinuo; Iguchi, Aiko; Liu, Mingming; Wang, Guanbo; Zhou, Mo; Vudriko, Patrick; Efstratiou, Artemis; Changbunjong, Tanasak; Sungpradit, Sivapong; Ratanakorn, Parntep; Moonarmart, Walasinee; Sedwisai, Poonyapat; Weluwanarak, Thekhawet; Wongsawang, Witsanu; Suzuki, Hiroshi; Xuan, Xuenan

    2017-02-01

    Babesia spp., Theileria orientalis, and Anaplasma marginale are significant tick-borne pathogens that affect the health and productivity of cattle in tropical and subtropical areas. In this study, we used PCR to detect the presence of Babesia bovis, Babesia bigemina, and T. orientalis in 279 beef cattle from Western Thailand and A. marginale in 608 beef cattle from the north, northeastern, and western regions. The PCRs were performed using species-specific primers based on the B. bovis spherical body protein 2 (BboSBP2), B. bigemina rhoptry-associated protein 1a (BbiRAP-1a), T. orientalis major piroplasm surface protein (ToMPSP), and A. marginale major surface protein 4 (AmMSP4) genes. To determine the genetic diversity of the above parasites, amplicons of B. bovis and B. bigemina ITS1-5.8s rRNA gene-ITS2 regions (B. bovis ITS, B. bigemina ITS), ToMPSP, and AmMSP4 genes were sequenced for phylogenetic analysis. PCR results revealed that the prevalence of B. bovis, B. bigemina, T. orientalis, and A. marginale in the Western region was 11.1, 12.5, 7.8, and 39.1 %, respectively. Coinfections of two or three parasites were observed in 17.9 % of the animals sampled. The study revealed that the prevalence of A. marginale in the western region was higher than in the north and northeastern regions (7 %). Sequence analysis showed the BboSBP2 gene to be more conserved than B. bovis ITS in the different isolates and, similarly, the BbiRAP-1a was more conserved than B. bigemina ITS. In the phylogenetic analysis, T. orientalis MPSP sequences were classified into types 3, 5, and 7 as previously reported. A. marginale MSP4 gene sequences shared high identity and similarity with each other and clustered with isolates from other countries. This study provides information on the prevalence and genetic diversity of tick-borne pathogens in beef cattle and highlights the need for effective strategies to control these pathogens in Thailand.

  5. Genetic diversity and molecular phylogeny of Anaplasma marginale studied longitudinally under natural transmission conditions in Rio de Janeiro, Brazil.

    PubMed

    Silva, Jenevaldo Barbosa; Gonçalves, Luiz Ricardo; Varani, Alessandro de Mello; André, Marcos Rogério; Machado, Rosangela Zacarias

    2015-06-01

    Anaplasma marginale is the most prevalent tick-borne pathogen in cattle in tropical and subtropical regions of the world. Major Surface Protein 1a (MSP1a) has been found to be a stable genetic marker for identifying A. marginale isolates within geographical regions. It is conserved in cattle during infection and tick-borne transmission of the pathogen. The aim of the present longitudinal study was to determine occurrences of genetic diversity associated with high prevalence of A. marginale under natural transmission conditions. Twenty calves were evaluated every 3 months during the first year of life. Rickettsemia levels due to A. marginale, measured as the number of msp1αcopies/ml in the blood of positive calves, ranged from 2.06×10(3) to 4.36×10(12). The numbers of MSP1a tandem repeats among MSP1a tandem repeats were 3 and 6. The predominant msp1α microsatellite was E, and another MSP1a tandem repeat was found that presented genotype G. Nineteen different MSP1a tandem repeats of A. marginale were found circulating in animals. The MSP1a tandem repeats 4-63-27 (27.5%), 78-24(2)-25-31 (n=21.6%) and τ-10(2)-15 (n=17.6%) were the ones most commonly observed. Twenty-two MSP1a tandem repeats resulted in new sequences with amino acid changes, as shown in this study. Thirty sequences were found for the first time in Brazil. Glycine, glutamate, serine and alanine amino acids were found at position 20. During the study, 80% (16/20) of the animals were infected by more than one genotype. Three animals were born infected, with MSP1a tandem repeats 4-63-27, 78-24(2)-25-31 and τ-10(2)-15, thus indicating occurrence of transplacental transmission. In the phylogenetic analysis, 19 different MSP1a tandem repeats of A. marginale were found in the cattle, which suggested that many MSP1a tandem repeats and high variation in MSP1a were occurring.

  6. Differential expression of genes in salivary glands of male Rhipicephalus (Boophilus)microplus in response to infection with Anaplasma marginale.

    PubMed

    Zivkovic, Zorica; Esteves, Eliane; Almazán, Consuelo; Daffre, Sirlei; Nijhof, Ard M; Kocan, Katherine M; Jongejan, Frans; de la Fuente, José

    2010-03-18

    Bovine anaplasmosis, caused by the rickettsial tick-borne pathogen Anaplasma marginale (Rickettsiales: Anaplasmataceae), is vectored by Rhipicephalus (Boophilus)microplus in many tropical and subtropical regions of the world. A. marginale undergoes a complex developmental cycle in ticks which results in infection of salivary glands from where the pathogen is transmitted to cattle. In previous studies, we reported modification of gene expression in Dermacentor variabilis and cultured Ixodes scapularis tick cells in response to infection with A. marginale. In these studies, we extended these findings by use of a functional genomics approach to identify genes differentially expressed in R. microplus male salivary glands in response to A. marginale infection. Additionally, a R. microplus-derived cell line, BME26, was used for the first time to also study tick cell gene expression in response to A. marginale infection. Suppression subtractive hybridization libraries were constructed from infected and uninfected ticks and used to identify genes differentially expressed in male R. microplus salivary glands infected with A. marginale. A total of 279 ESTs were identified as candidate differentially expressed genes. Of these, five genes encoding for putative histamine-binding protein (22Hbp), von Willebrand factor (94Will), flagelliform silk protein (100Silk), Kunitz-like protease inhibitor precursor (108Kunz) and proline-rich protein BstNI subfamily 3 precursor (7BstNI3) were confirmed by real-time RT-PCR to be down-regulated in tick salivary glands infected with A. marginale. The impact of selected tick genes on A. marginale infections in tick salivary glands and BME26 cells was characterized by RNA interference. Silencing of the gene encoding for putative flagelliform silk protein (100Silk) resulted in reduced A. marginale infection in both tick salivary glands and cultured BME26 cells, while silencing of the gene encoding for subolesin (4D8) significantly reduced

  7. Differential expression of genes in salivary glands of male Rhipicephalus (Boophilus)microplus in response to infection with Anaplasma marginale

    PubMed Central

    2010-01-01

    Background Bovine anaplasmosis, caused by the rickettsial tick-borne pathogen Anaplasma marginale (Rickettsiales: Anaplasmataceae), is vectored by Rhipicephalus (Boophilus)microplus in many tropical and subtropical regions of the world. A. marginale undergoes a complex developmental cycle in ticks which results in infection of salivary glands from where the pathogen is transmitted to cattle. In previous studies, we reported modification of gene expression in Dermacentor variabilis and cultured Ixodes scapularis tick cells in response to infection with A. marginale. In these studies, we extended these findings by use of a functional genomics approach to identify genes differentially expressed in R. microplus male salivary glands in response to A. marginale infection. Additionally, a R. microplus-derived cell line, BME26, was used for the first time to also study tick cell gene expression in response to A. marginale infection. Results Suppression subtractive hybridization libraries were constructed from infected and uninfected ticks and used to identify genes differentially expressed in male R. microplus salivary glands infected with A. marginale. A total of 279 ESTs were identified as candidate differentially expressed genes. Of these, five genes encoding for putative histamine-binding protein (22Hbp), von Willebrand factor (94Will), flagelliform silk protein (100Silk), Kunitz-like protease inhibitor precursor (108Kunz) and proline-rich protein BstNI subfamily 3 precursor (7BstNI3) were confirmed by real-time RT-PCR to be down-regulated in tick salivary glands infected with A. marginale. The impact of selected tick genes on A. marginale infections in tick salivary glands and BME26 cells was characterized by RNA interference. Silencing of the gene encoding for putative flagelliform silk protein (100Silk) resulted in reduced A. marginale infection in both tick salivary glands and cultured BME26 cells, while silencing of the gene encoding for subolesin (4D8

  8. Identification of a vertically transmitted strain from Anaplasma marginale (UFMG3): Molecular and phylogenetic characterization, and evaluation of virulence.

    PubMed

    Silvestre, Bruna T; Silveira, Júlia A G; Meneses, Rodrigo M; Facury-Filho, Elias J; Carvalho, Antônio U; Ribeiro, Múcio F B

    2016-02-01

    Bovine anaplasmosis is a disease caused by the intraerythrocytic rickettsia species Anaplasma marginale and results in great economic losses in tropical and subtropical regions. Vertical transmission is an important phenomenon that contributes to the persistence of different strains of the agent within the same herd. The identification of new strains and genetic characterization studies are essential to understanding their epidemiology and virulence and for vaccine development. The aim of this study was to perform molecular and phylogenetic characterizations of a new vertically transmitted strain from A. marginale and to evaluate its virulence by experimental inoculation of rickettsia-free calves. Thirty newborn Holstein calves were subjected to molecular tests for the detection of A. marginale, Babesia bovis and Babesia bigemina. Calves positive for A. marginale (n=3) were splenectomized and monitored for the clinical manifestations of anaplasmosis. Blood samples from one of the calves that presented rickettsemia of 42.8% and spontaneous recovery of clinical parameters were used for molecular and phylogenetic characterization (msp1a gene), and inoculum production was used for the evaluation of virulence. This strain was identified as UFMG3. Three tandem repeat forms (13 and MGI19) were identified from the analysis of the msp1a gene, in which the form MGI19 appeared twice. Analysis of these repeats revealed the presence of the sequences QASTSS and SSASGQQQESS and of aspartic acid (D) at position 20 of both repeats. Phylogenetic analysis showed a close relationship among the UFMG3, MGI19 and UFMG2 strains. For virulence evaluation, six Holstein calves were inoculated intravenously with 2×10(7)A. marginale UFMG3-infected erythrocytes. The calves showed maximum rickettsemia of 5.1%, a moderate decrease in packed cell volume and spontaneous recovery of clinical parameters without the need for treatment. The results of experimental inoculation suggest that the strain A

  9. Large size space construction for space exploitation

    NASA Astrophysics Data System (ADS)

    Kondyurin, Alexey

    2016-07-01

    Space exploitation is impossible without large space structures. We need to make sufficient large volume of pressurized protecting frames for crew, passengers, space processing equipment, & etc. We have to be unlimited in space. Now the size and mass of space constructions are limited by possibility of a launch vehicle. It limits our future in exploitation of space by humans and in development of space industry. Large-size space construction can be made with using of the curing technology of the fibers-filled composites and a reactionable matrix applied directly in free space. For curing the fabric impregnated with a liquid matrix (prepreg) is prepared in terrestrial conditions and shipped in a container to orbit. In due time the prepreg is unfolded by inflating. After polymerization reaction, the durable construction can be fitted out with air, apparatus and life support systems. Our experimental studies of the curing processes in the simulated free space environment showed that the curing of composite in free space is possible. The large-size space construction can be developed. A project of space station, Moon base, Mars base, mining station, interplanet space ship, telecommunication station, space observatory, space factory, antenna dish, radiation shield, solar sail is proposed and overviewed. The study was supported by Humboldt Foundation, ESA (contract 17083/03/NL/SFe), NASA program of the stratospheric balloons and RFBR grants (05-08-18277, 12-08-00970 and 14-08-96011).

  10. Exploiting Allee effects for managing biological invasions.

    PubMed

    Tobin, Patrick C; Berec, Luděk; Liebhold, Andrew M

    2011-06-01

    Biological invasions are a global and increasing threat to the function and diversity of ecosystems. Allee effects (positive density dependence) have been shown to play an important role in the establishment and spread of non-native species. Although Allee effects can be considered a bane in conservation efforts, they can be a benefit in attempts to manage non-native species. Many biological invaders are subject to some form of an Allee effect, whether due to a need to locate mates, cooperatively feed or reproduce or avoid becoming a meal, yet attempts to highlight the specific exploitation of Allee effects in biological invasions are surprisingly unprecedented. In this review, we highlight current strategies that effectively exploit an Allee effect, and propose novel means by which Allee effects can be manipulated to the detriment of biological invaders. We also illustrate how the concept of Allee effects can be integral in risk assessments and in the prioritization of resources allocated to manage non-native species, as some species beset by strong Allee effects could be less successful as invaders. We describe how tactics that strengthen an existing Allee effect or create new ones could be used to manage biological invasions more effectively.

  11. Exploration versus exploitation in polydomous ant colonies.

    PubMed

    Cook, Zoe; Franks, Daniel W; Robinson, Elva J H

    2013-04-21

    In socially foraging species resource information can be shared between individuals, increasing foraging success. In ant colonies, nestmate recruitment allows high exploitation rates at known resources however, to maximise foraging efficiency this must be balanced with searching for new resources. Many ant species form colonies inhabiting two or more spatially separated but socially connected nests: this type of organisation is known as polydomy. Polydomous colonies may benefit from increased foraging efficiency by carrying out dispersed-central place foraging. However, decentralisation of the colony may affect recruitment success by limiting interaction between ants based in separate nests. We use an agent-based model which compares the foraging success of monodomous and polydomous colonies in different food environments, incorporating recruitment through pheromone trails and group foraging. In contrast to previous results we show that polydomy is beneficial in some but not all cases. Polydomous colonies discover resources at a higher rate, making them more successful when food is highly dispersed, but their relative success can be lowered by limitations on recruitment success. Monodomous colonies can have higher foraging efficiency than polydomous colonies by exploiting food more rapidly. The results show the importance of interactions between recruitment strategy, colony size, and colony organisation. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. New stratigraphic technologies in exploration/exploitation

    SciTech Connect

    Scott, R.W.; Lane, H.R.

    1995-09-01

    With the demand for increased accuracy and precision in the interpretation of seismic sections for the prediction of reservoirs and traps, traditional paleontologic and new non-paleontologic stratigraphic data have assumed a greater role in exploration/exploitation. Strata bounded by sequence boundaries can be dated have assumed a greater role in exploration/exploitation. Strata bounded by sequence boundaries can be dated precisely by the graphic correlation technique. For example, Arabian Cretaceous reservoirs pinch out by facies change and diagenesis into coeval strata. But Pliocene sands in the Gulf Coast are truncated by regional unconformities accentuated by salt dome movement. In this Plio-Pleistocene section high precision chronostratigraphy is achieved by the integration of paleontological and geochemical data. Detailed biostratigraphic analysis of many sections of Mississippian carbonates along the eastern side of the Transcontinental Arch allows for the recognition of four unconformity-bounded units (biothems of Lane and others, 1994). These biotherms have been traced form Illinois to New Mexico and are not easily detected in the subsurface by using seismic or electric log technology because of the rather uniform carbonate lithologies composing the entire Mississippian in the region. Mississippian hydrocarbon production in western Kansas can be shown to be associated with these intra-Mississippian carbonate-on-carbonate unconformities that are easily detected biostratigraphically.

  13. Sentinel-1 mission scientific exploitation activities

    NASA Astrophysics Data System (ADS)

    Desnos, Yves louis; Foumelis, Michael; Engdahl, Marcus

    2017-04-01

    The Sentinel-1 mission is the European Imaging Radar Observatory for the Copernicus joint initiative of the European Commission (EC) and the European Space Agency (ESA). Sentinel-1 mission is composed of a constellation of two satellites, Sentinel-1A and Sentinel-1B (launched in April 2014 and April 2016, respectively), sharing the same orbital plane and featuring a short repeat cycle of 6 days optimised for Synthetic Aperture Radar (SAR) interferometry science and applications. The full operation capacity was achieved after the completion of the Sentinel-1B in-orbit commissioning on 14 September 2016. Sentinel-1 data are freely available via the ESA's Sentinels Scientific Data Hub since October 2014. The data uptake by the science community has been unprecedented and numerous results have been published to date. The objective of the current paper is to provide a brief overview of the latest ESA activities, in the frame of the Scientific Exploitation of Operational Missions (SEOM) programme, aimed to facilitate the scientific exploitation of Sentinel-1 mission as well as discuss future opportunities for research.

  14. Compressed sensing MRI exploiting complementary dual decomposition.

    PubMed

    Park, Suhyung; Park, Jaeseok

    2014-04-01

    Compressed sensing (CS) MRI exploits the sparsity of an image in a transform domain to reconstruct the image from incoherently under-sampled k-space data. However, it has been shown that CS suffers particularly from loss of low-contrast image features with increasing reduction factors. To retain image details in such degraded experimental conditions, in this work we introduce a novel CS reconstruction method exploiting feature-based complementary dual decomposition with joint estimation of local scale mixture (LSM) model and images. Images are decomposed into dual block sparse components: total variation for piecewise smooth parts and wavelets for residuals. The LSM model parameters of residuals in the wavelet domain are estimated and then employed as a regional constraint in spatially adaptive reconstruction of high frequency subbands to restore image details missing in piecewise smooth parts. Alternating minimization of the dual image components subject to data consistency is performed to extract image details from residuals and add them back to their complementary counterparts while the LSM model parameters and images are jointly estimated in a sequential fashion. Simulations and experiments demonstrate the superior performance of the proposed method in preserving low-contrast image features even at high reduction factors.

  15. Evidence of the importance of host habitat use in predicting the dilution effect of wild boar for deer exposure to Anaplasma spp.

    PubMed

    Estrada-Peña, Agustín; Acevedo, Pelayo; Ruiz-Fons, Francisco; Gortázar, Christian; de la Fuente, José

    2008-08-20

    Foci of tick-borne pathogens occur at fine spatial scales, and depend upon a complex arrangement of factors involving climate, host abundance and landscape composition. It has been proposed that the presence of hosts that support tick feeding but not pathogen multiplication may dilute the transmission of the pathogen. However, models need to consider the spatial component to adequately explain how hosts, ticks and pathogens are distributed into the landscape. In this study, a novel, lattice-derived, behavior-based, spatially-explicit model was developed to test how changes in the assumed perception of different landscape elements affect the outcome of the connectivity between patches and therefore the dilution effect. The objective of this study was to explain changes in the exposure rate (ER) of red deer to Anaplasma spp. under different configurations of suitable habitat and landscape fragmentation in the presence of variable densities of the potentially diluting host, wild boar. The model showed that the increase in habitat fragmentation had a deep impact on Habitat Sharing Ratio (HSR), a parameter describing the amount of habitat shared by red deer and wild boar, weighted by the probability of the animals to remain together in the same patch (according to movement rules), the density of ticks and the density of animals at a given vegetation patch, and decreased the dilution effect of wild boar on deer Anaplasma ER. The model was validated with data collected on deer, wild boar and tick densities, climate, landscape composition, host vegetation preferences and deer seropositivity to Anaplasma spp. (as a measure of ER) in 10 study sites in Spain. However, although conditions were appropriate for a dilution effect, empirical results did not show a decrease in deer ER in sites with high wild boar densities. The model showed that the HSR was the most effective parameter to explain the absence of the dilution effect. These results suggest that host habitat usage may

  16. Phylogeographic analysis reveals association of tick-borne pathogen, Anaplasma marginale, MSP1a sequences with ecological traits affecting tick vector performance

    PubMed Central

    Estrada-Peña, Agustín; Naranjo, Victoria; Acevedo-Whitehouse, Karina; Mangold, Atilio J; Kocan, Katherine M; de la Fuente, José

    2009-01-01

    Background The tick-borne pathogen Anaplasma marginale, which is endemic worldwide, is the type species of the genus Anaplasma (Rickettsiales: Anaplasmataceae). Rhipicephalus (Boophilus) microplus is the most important tick vector of A. marginale in tropical and subtropical regions of the world. Despite extensive characterization of the genetic diversity in A. marginale geographic strains using major surface protein sequences, little is known about the biogeography and evolution of A. marginale and other Anaplasma species. For A. marginale, MSP1a was shown to be involved in vector-pathogen and host-pathogen interactions and to have evolved under positive selection pressure. The MSP1a of A. marginale strains differs in molecular weight because of a variable number of tandem 23-31 amino acid repeats and has proven to be a stable marker of strain identity. While phylogenetic studies of MSP1a repeat sequences have shown evidence of A. marginale-tick co-evolution, these studies have not provided phylogeographic information on a global scale because of the high level of MSP1a genetic diversity among geographic strains. Results In this study we showed that the phylogeography of A. marginale MSP1a sequences is associated with world ecological regions (ecoregions) resulting in different evolutionary pressures and thence MSP1a sequences. The results demonstrated that the MSP1a first (R1) and last (RL) repeats and microsatellite sequences were associated with world ecoregion clusters with specific and different environmental envelopes. The evolution of R1 repeat sequences was found to be under positive selection. It is hypothesized that the driving environmental factors regulating tick populations could act on the selection of different A. marginale MSP1a sequence lineages, associated to each ecoregion. Conclusion The results reported herein provided the first evidence that the evolution of A. marginale was linked to ecological traits affecting tick vector performance. These

  17. Evidence of the Importance of Host Habitat Use in Predicting the Dilution Effect of Wild Boar for Deer Exposure to Anaplasma spp

    PubMed Central

    Estrada-Peña, Agustín; Acevedo, Pelayo; Ruiz-Fons, Francisco; Gortázar, Christian; de la Fuente, José

    2008-01-01

    Foci of tick-borne pathogens occur at fine spatial scales, and depend upon a complex arrangement of factors involving climate, host abundance and landscape composition. It has been proposed that the presence of hosts that support tick feeding but not pathogen multiplication may dilute the transmission of the pathogen. However, models need to consider the spatial component to adequately explain how hosts, ticks and pathogens are distributed into the landscape. In this study, a novel, lattice-derived, behavior-based, spatially-explicit model was developed to test how changes in the assumed perception of different landscape elements affect the outcome of the connectivity between patches and therefore the dilution effect. The objective of this study was to explain changes in the exposure rate (ER) of red deer to Anaplasma spp. under different configurations of suitable habitat and landscape fragmentation in the presence of variable densities of the potentially diluting host, wild boar. The model showed that the increase in habitat fragmentation had a deep impact on Habitat Sharing Ratio (HSR), a parameter describing the amount of habitat shared by red deer and wild boar, weighted by the probability of the animals to remain together in the same patch (according to movement rules), the density of ticks and the density of animals at a given vegetation patch, and decreased the dilution effect of wild boar on deer Anaplasma ER. The model was validated with data collected on deer, wild boar and tick densities, climate, landscape composition, host vegetation preferences and deer seropositivity to Anaplasma spp. (as a measure of ER) in 10 study sites in Spain. However, although conditions were appropriate for a dilution effect, empirical results did not show a decrease in deer ER in sites with high wild boar densities. The model showed that the HSR was the most effective parameter to explain the absence of the dilution effect. These results suggest that host habitat usage may

  18. Early Warning and Prediction of Interest Attacks and Exploits

    DTIC Science & Technology

    2005-09-01

    EARLY WARNING AND PREDICTION OF INTERNET ATTACKS AND EXPLOITS THESIS Brian P. Zeitz... EARLY WARNING AND PREDICTION OF INTERNET ATTACKS AND EXPLOITS THESIS Presented to the Faculty Department of Electrical and Computer...RELEASE; DISTRIBUTION UNLIMITED iv AFIT/GIA/ENG/05-06 EARLY WARNING AND PREDICTION OF INTERNET ATTACKS AND EXPLOITS

  19. Virtual Exploitation Environment Demonstration for Atmospheric Missions

    NASA Astrophysics Data System (ADS)

    Natali, Stefano; Mantovani, Simone; Hirtl, Marcus; Santillan, Daniel; Triebnig, Gerhard; Fehr, Thorsten; Lopes, Cristiano

    2017-04-01

    The scientific and industrial communities are being confronted with a strong increase of Earth Observation (EO) satellite missions and related data. This is in particular the case for the Atmospheric Sciences communities, with the upcoming Copernicus Sentinel-5 Precursor, Sentinel-4, -5 and -3, and ESA's Earth Explorers scientific satellites ADM-Aeolus and EarthCARE. The challenge is not only to manage the large volume of data generated by each mission / sensor, but to process and analyze the data streams. Creating synergies among the different datasets will be key to exploit the full potential of the available information. As a preparation activity supporting scientific data exploitation for Earth Explorer and Sentinel atmospheric missions, ESA funded the "Technology and Atmospheric Mission Platform" (TAMP) [1] [2] project; a scientific and technological forum (STF) has been set-up involving relevant European entities from different scientific and operational fields to define the platforḿs requirements. Data access, visualization, processing and download services have been developed to satisfy useŕs needs; use cases defined with the STF, such as study of the SO2 emissions for the Holuhraun eruption (2014) by means of two numerical models, two satellite platforms and ground measurements, global Aerosol analyses from long time series of satellite data, and local Aerosol analysis using satellite and LIDAR, have been implemented to ensure acceptance of TAMP by the atmospheric sciences community. The platform pursues the "virtual workspace" concept: all resources (data, processing, visualization, collaboration tools) are provided as "remote services", accessible through a standard web browser, to avoid the download of big data volumes and for allowing utilization of provided infrastructure for computation, analysis and sharing of results. Data access and processing are achieved through standardized protocols (WCS, WPS). As evolution toward a pre

  20. Development and assessment of a latex agglutination test based on recombinant MSP5 to detect antibodies against Anaplasma marginale in cattle.

    PubMed

    Ramos, Carlos A N; Araújo, Flábio R; Santos, Rafaelle C; Melo, Elaine S P; Sousa, Letícia C; Vidal, Carlos E S; Guerra, Neurisvan R; Ramos, Rafael A N

    2014-01-01

    The recombinant protein MSP5 has been established as an important antigen for serological diagnosis of Anaplasma marginale by enzyme-linked immunosorbent assay (ELISA). However, due to the high cost of specialized equipment, this technique is not accessible to all laboratories, especially in developing countries in areas where the disease is endemic. The present study describes the standardization of a latex agglutination test (LAT) to detect antibodies against A. marginale based on recombinant MSP5. Compared with indirect enzyme-linked immunosorbent assay (iELISA), the relative sensitivity and specificity of the LAT were 95.21% and 91.86% respectively, with an almost perfect agreement between tests (kappa index = 0.863). These results can be considered important for the serological diagnosis of A. marginale, as they indicate that the test represents a rapid and low cost alternative to ELISA.

  1. Active surveillance of Anaplasma marginale in populations of arthropod vectors (Acari: Ixodidae; Diptera: Tabanidae) during and after an outbreak of bovine anaplasmosis in southern Manitoba, Canada

    PubMed Central

    Yunik, Matthew E.M.; Galloway, Terry D.; Lindsay, L. Robbin

    2016-01-01

    Bovine anaplasmosis is the disease caused by the bacterium Anaplasma marginale. It can cause production loss and death in cattle and bison. This was a reportable disease in Canada until April 2014. Before then, infected herds were quarantined and culled, removing infected animals. In North America, A. marginale is biologically vectored by hard ticks (Acari: Ixodidae), Dermacentor variabilis and D. andersoni. Biting flies, particularly horse flies (Diptera: Tabanidae), can also act as mechanical vectors. An outbreak of bovine anaplasmosis, consisting of 14 herds, was detected in southern Manitoba in 2008. This outbreak lasted multiple rounds of testing and culling before eradication in 2011, suggesting local maintenance of the pathogen was occurring. We applied novel approaches to examine the vector ecology of this disease in this region. We did not detect A. marginale by screening of 2056 D. variabilis (2011 and 2012) and 520 horse flies (2011) using polymerase chain reaction (PCR). PMID:27127345

  2. Development and assessment of a latex agglutination test based on recombinant MSP5 to detect antibodies against Anaplasma marginale in cattle

    PubMed Central

    Ramos, Carlos A.N.; Araújo, Flábio R.; Santos, Rafaelle C.; Melo, Elaine S.P.; Sousa, Letícia C.; Vidal, Carlos E.S.; Guerra, Neurisvan R.; Ramos, Rafael A.N.

    2014-01-01

    The recombinant protein MSP5 has been established as an important antigen for serological diagnosis of Anaplasma marginale by enzyme-linked immunosorbent assay (ELISA). However, due to the high cost of specialized equipment, this technique is not accessible to all laboratories, especially in developing countries in areas where the disease is endemic. The present study describes the standardization of a latex agglutination test (LAT) to detect antibodies against A. marginale based on recombinant MSP5. Compared with indirect enzyme-linked immunosorbent assay (iELISA), the relative sensitivity and specificity of the LAT were 95.21% and 91.86% respectively, with an almost perfect agreement between tests (kappa index = 0.863). These results can be considered important for the serological diagnosis of A. marginale, as they indicate that the test represents a rapid and low cost alternative to ELISA. PMID:24948931

  3. Active surveillance of Anaplasma marginale in populations of arthropod vectors (Acari: Ixodidae; Diptera: Tabanidae) during and after an outbreak of bovine anaplasmosis in southern Manitoba, Canada.

    PubMed

    Yunik, Matthew E M; Galloway, Terry D; Lindsay, L Robbin

    2016-04-01

    Bovine anaplasmosis is the disease caused by the bacterium Anaplasma marginale. It can cause production loss and death in cattle and bison. This was a reportable disease in Canada until April 2014. Before then, infected herds were quarantined and culled, removing infected animals. In North America, A. marginale is biologically vectored by hard ticks (Acari: Ixodidae), Dermacentor variabilis and D. andersoni. Biting flies, particularly horse flies (Diptera: Tabanidae), can also act as mechanical vectors. An outbreak of bovine anaplasmosis, consisting of 14 herds, was detected in southern Manitoba in 2008. This outbreak lasted multiple rounds of testing and culling before eradication in 2011, suggesting local maintenance of the pathogen was occurring. We applied novel approaches to examine the vector ecology of this disease in this region. We did not detect A. marginale by screening of 2056 D. variabilis (2011 and 2012) and 520 horse flies (2011) using polymerase chain reaction (PCR).

  4. Exploiting epigenetic vulnerabilities for cancer therapeutics.

    PubMed

    Mair, Barbara; Kubicek, Stefan; Nijman, Sebastian M B

    2014-03-01

    Epigenetic deregulation is a hallmark of cancer, and there has been increasing interest in therapeutics that target chromatin-modifying enzymes and other epigenetic regulators. The rationale for applying epigenetic drugs to treat cancer is twofold. First, epigenetic changes are reversible, and drugs could therefore be used to restore the normal (healthy) epigenetic landscape. However, it is unclear whether drugs can faithfully restore the precancerous epigenetic state. Second, chromatin regulators are often mutated in cancer, making them attractive drug targets. However, in most instances it is unknown whether cancer cells are addicted to these mutated chromatin proteins, or whether their mutation merely results in epigenetic instability conducive to the selection of secondary aberrations. An alternative incentive for targeting chromatin regulators is the exploitation of cancer-specific vulnerabilities, including synthetic lethality, caused by epigenetic deregulation. We review evidence for the hypothesis that mechanisms other than oncogene addiction are a basis for the application of epigenetic drugs, and propose future research directions.

  5. Redressing China's strategy of water resource exploitation.

    PubMed

    Ran, Lishan; Lu, Xi Xi

    2013-03-01

    China, with the confrontation of water-related problems as an element of its long history, has been investing heavily in water engineering projects over the past few decades based on the assumption that these projects can solve its water problems. However, the anticipated benefits did not really occur, or at least not as large as expected. Instead, the results involved additional frustrations, such as biodiversity losses and human-induced disasters (i.e., landslides and earthquakes). Given its inherent shortcomings, the present engineering-dominated strategy for the management of water resources cannot help solve China's water problems and achieve its goal of low-carbon transformation. Therefore, the present strategy for water resources exploitation needs to be reevaluated and redressed. A policy change to achieve better management of Chinese rivers is urgently needed.

  6. Digital video steganalysis exploiting collusion sensitivity

    NASA Astrophysics Data System (ADS)

    Budhia, Udit; Kundur, Deepa

    2004-09-01

    In this paper we present an effective steganalyis technique for digital video sequences based on the collusion attack. Steganalysis is the process of detecting with a high probability and low complexity the presence of covert data in multimedia. Existing algorithms for steganalysis target detecting covert information in still images. When applied directly to video sequences these approaches are suboptimal. In this paper, we present a method that overcomes this limitation by using redundant information present in the temporal domain to detect covert messages in the form of Gaussian watermarks. Our gains are achieved by exploiting the collusion attack that has recently been studied in the field of digital video watermarking, and more sophisticated pattern recognition tools. Applications of our scheme include cybersecurity and cyberforensics.

  7. Explosives Detection: Exploitation of the Physical Signatures

    NASA Astrophysics Data System (ADS)

    Atkinson, David

    2010-10-01

    Explosives based terrorism is an ongoing threat that is evolving with respect to implementation, configuration and materials used. There are a variety of devices designed to detect explosive devices, however, each technology has limitations and operational constraints. A full understanding of the signatures available for detection coupled with the array of detection choices can be used to develop a conceptual model of an explosives screening operation. Physics based sensors provide a robust approach to explosives detection, typically through the identification of anomalies, and are currently used for screening in airports around the world. The next generation of detectors for explosives detection will need to be more sensitive and selective, as well as integrate seamlessly with devices focused on chemical signatures. An appreciation for the details of the physical signature exploitation in cluttered environments with time, space, and privacy constraints is necessary for effective explosives screening of people, luggage, cargo, and vehicles.

  8. Iron and Zinc Exploitation during Bacterial Pathogenesis

    PubMed Central

    Ma, Li; Terwilliger, Austen; Maresso, Anthony W.

    2016-01-01

    Ancient bacteria originated from metal-rich environments. Billions of years of evolution directed these tiny single cell creatures to exploit the versatile properties of metals in catalyzing chemical reactions and biological responses. The result is an entire metallome of proteins that use metal co-factors to facilitate key cellular process that range from the production of energy to the replication of DNA. Two key metals in this regard are iron and zinc, both abundant on Earth but not readily accessible in a human host. Instead, pathogenic bacteria must employ clever ways to acquire these metals. In this review we describe the many elegant ways these bacteria mine, regulate, and craft the use of two key metals (iron and zinc) to build a virulence arsenal that challenges even the most sophisticated immune response. PMID:26497057

  9. Are Youths' Feelings of Entitlement Always "Bad"?: Evidence for a Distinction between Exploitive and Non-Exploitive Dimensions of Entitlement

    ERIC Educational Resources Information Center

    Lessard, Jared; Greenberger, Ellen; Chen, Chuansheng; Farruggia, Susan

    2011-01-01

    Previous personality research (e.g., Campbell et al., 2004) has described the sense of entitlement as an unifactorial construct. In this study, we examined characteristics of two potential facets of entitlement: exploitive entitlement, characterized by exploitive interactions and expectations of special treatment, and non-exploitive entitlement,…

  10. Are Youths' Feelings of Entitlement Always "Bad"?: Evidence for a Distinction between Exploitive and Non-Exploitive Dimensions of Entitlement

    ERIC Educational Resources Information Center

    Lessard, Jared; Greenberger, Ellen; Chen, Chuansheng; Farruggia, Susan

    2011-01-01

    Previous personality research (e.g., Campbell et al., 2004) has described the sense of entitlement as an unifactorial construct. In this study, we examined characteristics of two potential facets of entitlement: exploitive entitlement, characterized by exploitive interactions and expectations of special treatment, and non-exploitive entitlement,…

  11. GOCE Exploitation for Moho Modeling and Applications

    NASA Astrophysics Data System (ADS)

    Sampierto, D.

    2011-07-01

    New ESA missions dedicated to the observation of the Earth from space, like the gravity-gradiometry mission GOCE and the radar altimetry mission CRYOSAT 2, foster research, among other subjects, also on inverse gravimetric problems and on the description of the nature and the geographical location of gravimetric signals. In this framework the GEMMA project (GOCE Exploitation for Moho Modeling and Applications), funded by the European Space Agency and Politecnico di Milano, aims at estimating the boundary between Earth's crust and mantle (the so called Mohorovičić discontinuity or Moho) from GOCE data in key regions of the world. In the project a solution based on a simple two layer model in spherical approximation is proposed. This inversion problem based on the linearization of the Newton's gravitational law around an approximate mean Moho surface will be solved by exploiting Wiener-Kolmogorov theory in the frequency domain where the depth of the Moho discontinuity will be treated as a random signal with a zero mean and its own covariance function. The algorithm can be applied in a numerically efficient way by using the Fast Fourier Transform. As for the gravity observations, we will consider grids of the anomalous gravitational potential and its second radial derivative at satellite altitude. In particular this will require first of all to elaborate GOCE data to obtain a local grid of the gravitational potential field and its second radial derivative and after that to separate the gravimetric signal due to the considered discontinuity from the gravitational effects of other geological structures present into the observations. The first problem can be solved by applying the so called space- wise approach to GOCE observations, while the second one can be achieved by considering a priori models and geophysical information by means of an appropriate Bayesan technique. Moreover other data such as ground gravity anomalies or seismic profiles can be combined, in an

  12. Mission Exploitation Platform PROBA-V

    NASA Astrophysics Data System (ADS)

    Goor, Erwin

    2016-04-01

    VITO and partners developed an end-to-end solution to drastically improve the exploitation of the PROBA-V EO-data archive (http://proba-v.vgt.vito.be/), the past mission SPOT-VEGETATION and derived vegetation parameters by researchers, service providers and end-users. The analysis of time series of data (+1PB) is addressed, as well as the large scale on-demand processing of near real-time data. From November 2015 an operational Mission Exploitation Platform (MEP) PROBA-V, as an ESA pathfinder project, will be gradually deployed at the VITO data center with direct access to the complete data archive. Several applications will be released to the users, e.g. - A time series viewer, showing the evolution of PROBA-V bands and derived vegetation parameters for any area of interest. - Full-resolution viewing services for the complete data archive. - On-demand processing chains e.g. for the calculation of N-daily composites. - A Virtual Machine will be provided with access to the data archive and tools to work with this data, e.g. various toolboxes and support for R and Python. After an initial release in January 2016, a research platform will gradually be deployed allowing users to design, debug and test applications on the platform. From the MEP PROBA-V, access to Sentinel-2 and landsat data will be addressed as well, e.g. to support the Cal/Val activities of the users. Users can make use of powerful Web based tools and can self-manage virtual machines to perform their work on the infrastructure at VITO with access to the complete data archive. To realise this, private cloud technology (openStack) is used and a distributed processing environment is built based on Hadoop. The Hadoop ecosystem offers a lot of technologies (Spark, Yarn, Accumulo, etc.) which we integrate with several open-source components. The impact of this MEP on the user community will be high and will completely change the way of working with the data and hence open the large time series to a larger

  13. Generating circuit tests by exploiting designed behavior

    NASA Astrophysics Data System (ADS)

    Shirley, Mark H.

    1988-12-01

    Generating tests for sequential devices is one of the hardest problems in designing and manufacturing digital circuits. This task is difficult primarily because internal components are accessible only indirectly, forcing a test generator to use the surrounding components collectively as a probe for detecting faults. This in turn forces the test generator to reason about complex interactions between the behaviors of these surrounding components. Current automated solutions are becoming ineffective as designs grow larger and more complex. Yet, despite the complexity, human experts remain remarkably successful, in part, because they use knowledge from many sources and use a variety of reasoning techniques. This thesis exploits several kinds of expert knowledge about circuits and test generation not used by the current algorithms. First, many test generation problems can be solved efficiently using operation relations, a novel representation of circuit behavior that connects internal component operations with directly executable circuit operations. Operation relations can be computed efficiently for sequential circuits that provide few operations at their interfaces by searching traces of simulated circuit behavior. Second, experts write test programs rather than test vectors because programs are a more readable and compact representation for tests than vectors are. Test programs can be constructed automatically by merging test program fragments using expert supplied goal-refined rules and domain independent planning techniques from artificial intelligence.

  14. The Spanish network for Gaia Science Exploitation

    NASA Astrophysics Data System (ADS)

    Figueras, F.; Jordi, C.; Luri, X.; Torra, J.; REG Executive Committee Team; Gaia UB Team

    2017-03-01

    The ''Red Española de Explotación Científica de Gaia'' (REG) continues to intensify its activities facing the imminent publication of the first and second Gaia data releases (14 September, 2016 and Q4-2017, respectively). The network, supported by the MINECO under contract Acciones de dinamizaci ´on, Redes de Excelencia (2016-2017), has as major priority the task to coordinate and support the collective activities developed by its more than 150 members. At present, REG plays a prominent role in the preparation of the Spanish community for the use of the Gaia data archive (a task lead by the Spanish team), in the work to exploit the Gaia-ESO survey collected during the last four years and in supporting the preparation of the science case and survey plan for WEAVE, the new multi-object spectrograph for the WHT at Canary Islands (commissioning, 2018). These activities are described together with the schedule of future national and international science meetings and the outreach activities being organized for the first and second Data Releases

  15. Examining the potential exploitation of UNOS policies.

    PubMed

    Zink, Sheldon; Wertlieb, Stacey; Catalano, John; Marwin, Victor

    2005-01-01

    The United Network for Organ Sharing (UNOS) waiting list was designed as a just and equitable system through which the limited number of organs is allocated to the millions of Americans in need of a transplant. People have trusted the system because of the belief that everyone on the list has an equal opportunity to receive an organ and also that allocation is blind to matters of financial standing, celebrity or political power. Recent events have revealed that certain practices and policies have the potential to be exploited. The policies addressed in this paper enable those on the list with the proper resources to gain an advantage over other less fortunate members, creating a system that benefits not the individual most in medical need, but the one with the best resources. These policies are not only unethical but threaten the balance and success of the entire UNOS system. This paper proposes one possible solution, which seeks to balance the concepts of justice and utility.

  16. Haploids in flowering plants: origins and exploitation.

    PubMed

    Dunwell, Jim M

    2010-05-01

    The first haploid angiosperm, a dwarf form of cotton with half the normal chromosome complement, was discovered in 1920, and in the ninety years since then such plants have been identified in many other species. They can occur either spontaneously or can be induced by modified pollination methods in vivo, or by in vitro culture of immature male or female gametophytes. Haploids represent an immediate, one-stage route to homozygous diploids and thence to F(1) hybrid production. The commercial exploitation of heterosis in such F(1) hybrids leads to the development of hybrid seed companies and subsequently to the GM revolution in agriculture. This review describes the range of techniques available for the isolation or induction of haploids and discusses their value in a range of areas, from fundamental research on mutant isolation and transformation, through to applied aspects of quantitative genetics and plant breeding. It will also focus on how molecular methods have been used recently to explore some of the underlying aspects of this fascinating developmental phenomenon.

  17. Relationship between exploitation, oscillation, MSY and extinction.

    PubMed

    Ghosh, Bapan; Kar, T K; Legovic, T

    2014-10-01

    We give answers to two important problems arising in current fisheries: (i) how maximum sustainable yield (MSY) policy is influenced by the initial population level, and (ii) how harvesting, oscillation and MSY are related to each other in prey-predator systems. To examine the impact of initial population on exploitation, we analyze a single species model with strong Allee effect. It is found that even when the MSY exists, the dynamic solution may not converge to the equilibrium stock if the initial population level is higher but near the critical threshold level. In a prey-predator system with Allee effect in the prey species, the initial population does not have such important impact neither on MSY nor on maximum sustainable total yield (MSTY). However, harvesting the top predator may cause extinction of all species if odd number of trophic levels exist in the ecosystem. With regard to the second problem, we study two prey-predator models and establish that increasing harvesting effort either on prey, predator or both prey and predator destroys previously existing oscillation. Moreover, equilibrium stock both at MSY and MSTY level is stable. We also discuss the validity of found results to other prey-predator systems.

  18. Exploiting spatial descriptions in visual scene analysis.

    PubMed

    Ziegler, Leon; Johannsen, Katrin; Swadzba, Agnes; De Ruiter, Jan P; Wachsmuth, Sven

    2012-08-01

    The reliable automatic visual recognition of indoor scenes with complex object constellations using only sensor data is a nontrivial problem. In order to improve the construction of an accurate semantic 3D model of an indoor scene, we exploit human-produced verbal descriptions of the relative location of pairs of objects. This requires the ability to deal with different spatial reference frames (RF) that humans use interchangeably. In German, both the intrinsic and relative RF are used frequently, which often leads to ambiguities in referential communication. We assume that there are certain regularities that help in specific contexts. In a first experiment, we investigated how speakers of German describe spatial relationships between different pieces of furniture. This gave us important information about the distribution of the RFs used for furniture-predicate combinations, and by implication also about the preferred spatial predicate. The results of this experiment are compiled into a computational model that extracts partial orderings of spatial arrangements between furniture items from verbal descriptions. In the implemented system, the visual scene is initially scanned by a 3D camera system. From the 3D point cloud, we extract point clusters that suggest the presence of certain furniture objects. We then integrate the partial orderings extracted from the verbal utterances incrementally and cumulatively with the estimated probabilities about the identity and location of objects in the scene, and also estimate the probable orientation of the objects. This allows the system to significantly improve both the accuracy and richness of its visual scene representation.

  19. Exploiting interspecific olfactory communication to monitor predators.

    PubMed

    Garvey, Patrick M; Glen, Alistair S; Clout, Mick N; Wyse, Sarah V; Nichols, Margaret; Pech, Roger P

    2017-03-01

    Olfaction is the primary sense of many mammals and subordinate predators use this sense to detect dominant species, thereby reducing the risk of an encounter and facilitating coexistence. Chemical signals can act as repellents or attractants and may therefore have applications for wildlife management. We devised a field experiment to investigate whether dominant predator (ferret Mustela furo) body odor would alter the behavior of three common mesopredators: stoats (Mustela erminea), hedgehogs (Erinaceus europaeus), and ship rats (Rattus rattus). We predicted that apex predator odor would lead to increased detections, and our results support this hypothesis as predator kairomones (interspecific olfactory messages that benefit the receiver) provoked "eavesdropping" behavior by mesopredators. Stoats exhibited the most pronounced responses, with kairomones significantly increasing the number of observations and the time spent at a site, so that their occupancy estimates changed from rare to widespread. Behavioral responses to predator odors can therefore be exploited for conservation and this avenue of research has not yet been extensively explored. A long-life lure derived from apex predator kairomones could have practical value, especially when there are plentiful resources that reduce the efficiency of food-based lures. Our results have application for pest management in New Zealand and the technique of using kairomones to monitor predators could have applications for conservation efforts worldwide.

  20. Exploiting intrinsic fluctuations to identify model parameters.

    PubMed

    Zimmer, Christoph; Sahle, Sven; Pahle, Jürgen

    2015-04-01

    Parameterisation of kinetic models plays a central role in computational systems biology. Besides the lack of experimental data of high enough quality, some of the biggest challenges here are identification issues. Model parameters can be structurally non-identifiable because of functional relationships. Noise in measured data is usually considered to be a nuisance for parameter estimation. However, it turns out that intrinsic fluctuations in particle numbers can make parameters identifiable that were previously non-identifiable. The authors present a method to identify model parameters that are structurally non-identifiable in a deterministic framework. The method takes time course recordings of biochemical systems in steady state or transient state as input. Often a functional relationship between parameters presents itself by a one-dimensional manifold in parameter space containing parameter sets of optimal goodness. Although the system's behaviour cannot be distinguished on this manifold in a deterministic framework it might be distinguishable in a stochastic modelling framework. Their method exploits this by using an objective function that includes a measure for fluctuations in particle numbers. They show on three example models, immigration-death, gene expression and Epo-EpoReceptor interaction, that this resolves the non-identifiability even in the case of measurement noise with known amplitude. The method is applied to partially observed recordings of biochemical systems with measurement noise. It is simple to implement and it is usually very fast to compute. This optimisation can be realised in a classical or Bayesian fashion.

  1. GOCE: mission accomplished but exploitation continues

    NASA Astrophysics Data System (ADS)

    Floberghagen, Rune; Fehringer, Michael; Steiger, Christoph; Romanazzo, Massimo; Frommknecht, Bjoern; Klinkrad, Heiner

    2014-05-01

    Launched in March 2009 into an extremely low Earth orbit on an originally 20-months mission to map the quasi-static gravity field, the GOCE satellite more than tripled its expected measurement return. Being the lowest orbiting research satellite since the beginning, GOCE underwent a series of orbit lowerings towards the end of its lifetime in order to further maximise the signal content (both in amplitude and scale) of the gravity gradient and hi-lo satellite-to-satellite observations. Gravity field mapping in drag-compensation mode at 224 km altitude was completed on 21 October 2013, after which the satellite rapidly decayed deeper into the upper layers of the atmosphere. During this phase all instruments and avionics equipment were acquiring data. Finally, the satellite re-entered over the South Atlantic ocean in the very early hours of 11 November 2013 (UTC). This contribution describes the science return of GOCE in its final phase, including the de-orbiting and re-entry phase. It provides an overview of the impact of GOCE in the earth sciences, incl. geodesy, oceanography, solid earth and aeronomy. Plans for continued exploitation of GOCE data - within and outside the context of the European Space Agency programmes - will be also presented.

  2. Exploiting core knowledge for visual object recognition.

    PubMed

    Schurgin, Mark W; Flombaum, Jonathan I

    2017-03-01

    Humans recognize thousands of objects, and with relative tolerance to variable retinal inputs. The acquisition of this ability is not fully understood, and it remains an area in which artificial systems have yet to surpass people. We sought to investigate the memory process that supports object recognition. Specifically, we investigated the association of inputs that co-occur over short periods of time. We tested the hypothesis that human perception exploits expectations about object kinematics to limit the scope of association to inputs that are likely to have the same token as a source. In several experiments we exposed participants to images of objects, and we then tested recognition sensitivity. Using motion, we manipulated whether successive encounters with an image took place through kinematics that implied the same or a different token as the source of those encounters. Images were injected with noise, or shown at varying orientations, and we included 2 manipulations of motion kinematics. Across all experiments, memory performance was better for images that had been previously encountered with kinematics that implied a single token. A model-based analysis similarly showed greater memory strength when images were shown via kinematics that implied a single token. These results suggest that constraints from physics are built into the mechanisms that support memory about objects. Such constraints-often characterized as 'Core Knowledge'-are known to support perception and cognition broadly, even in young infants. But they have never been considered as a mechanism for memory with respect to recognition. (PsycINFO Database Record

  3. Exploiting fungal cell wall components in vaccines

    PubMed Central

    Levitz, Stuart M.; Huang, Haibin; Ostroff, Gary R.; Specht, Charles A.

    2014-01-01

    Innate recognition of fungi leads to strong adaptive immunity. Investigators are trying to exploit this observation in vaccine development by combining antigens with evolutionarily conserved fungal cell wall carbohydrates to induce protective responses. Best studied is β-1,3-glucan, a glycan that activates complement and is recognized by Dectin-1. Administration of antigens in association with β-1,3-glucan, either by direct conjugation or complexed in glucan particles, results in robust humoral and cellular immune responses. While the host has a host of mannose receptors, responses to fungal mannoproteins generally are amplified if cells are cooperatively stimulated with an additional danger signal such as a toll-like receptor agonist. Chitosan, a polycationic homopolymer of glucosamine manufactured by the deacetylation of chitin, is being studied as an adjuvant in DNA and protein-based vaccines. It appears particularly promising in mucosal vaccines. Finally, universal and organism-specific fungal vaccines have been formulated by conjugating fungal cell wall glycans to carrier proteins. A major challenge will be to advance these experimental findings so that at risk patients can be protected. PMID:25404118

  4. Tonic Dopamine Modulates Exploitation of Reward Learning

    PubMed Central

    Beeler, Jeff A.; Daw, Nathaniel; Frazier, Cristianne R. M.; Zhuang, Xiaoxi

    2010-01-01

    The impact of dopamine on adaptive behavior in a naturalistic environment is largely unexamined. Experimental work suggests that phasic dopamine is central to reinforcement learning whereas tonic dopamine may modulate performance without altering learning per se; however, this idea has not been developed formally or integrated with computational models of dopamine function. We quantitatively evaluate the role of tonic dopamine in these functions by studying the behavior of hyperdopaminergic DAT knockdown mice in an instrumental task in a semi-naturalistic homecage environment. In this “closed economy” paradigm, subjects earn all of their food by pressing either of two levers, but the relative cost for food on each lever shifts frequently. Compared to wild-type mice, hyperdopaminergic mice allocate more lever presses on high-cost levers, thus working harder to earn a given amount of food and maintain their body weight. However, both groups show a similarly quick reaction to shifts in lever cost, suggesting that the hyperdominergic mice are not slower at detecting changes, as with a learning deficit. We fit the lever choice data using reinforcement learning models to assess the distinction between acquisition and expression the models formalize. In these analyses, hyperdopaminergic mice displayed normal learning from recent reward history but diminished capacity to exploit this learning: a reduced coupling between choice and reward history. These data suggest that dopamine modulates the degree to which prior learning biases action selection and consequently alters the expression of learned, motivated behavior. PMID:21120145

  5. Accelerating Large Data Analysis By Exploiting Regularities

    NASA Technical Reports Server (NTRS)

    Moran, Patrick J.; Ellsworth, David

    2003-01-01

    We present techniques for discovering and exploiting regularity in large curvilinear data sets. The data can be based on a single mesh or a mesh composed of multiple submeshes (also known as zones). Multi-zone data are typical to Computational Fluid Dynamics (CFD) simulations. Regularities include axis-aligned rectilinear and cylindrical meshes as well as cases where one zone is equivalent to a rigid-body transformation of another. Our algorithms can also discover rigid-body motion of meshes in time-series data. Next, we describe a data model where we can utilize the results from the discovery process in order to accelerate large data visualizations. Where possible, we replace general curvilinear zones with rectilinear or cylindrical zones. In rigid-body motion cases we replace a time-series of meshes with a transformed mesh object where a reference mesh is dynamically transformed based on a given time value in order to satisfy geometry requests, on demand. The data model enables us to make these substitutions and dynamic transformations transparently with respect to the visualization algorithms. We present results with large data sets where we combine our mesh replacement and transformation techniques with out-of-core paging in order to achieve significant speed-ups in analysis.

  6. Understanding Online Child Sexual Exploitation Offenses.

    PubMed

    Ly, Thanh; Murphy, Lisa; Fedoroff, J Paul

    2016-08-01

    In the past three decades, there has been an exponential increase in the worldwide availability of Internet access and devices that are able to access online materials. This literature review investigated whether increased accessibility of Internet child pornography (CP) increases the risk of in-person child sexual exploitation. The current review found little to no evidence that availability of the Internet has increased the worldwide incidence or prevalence of in-person child sexual abuse. In fact, during the time period in which the Internet has flourished, international crime statistics have shown a steady decrease of in-person child sexual abuse. The only exception to this trend is an increase in Internet child pornography or luring offenses (e.g., Stats Can, 2014), which involves child abuse by definition. This article reviews the impact of the Internet on child sexual abuse. It also reviews the characteristics of online CP offenders. Treatment of these offenders and prevention of such offenses is also discussed.

  7. Exploiting protein intrinsic flexibility in drug design.

    PubMed

    Lukman, Suryani; Verma, Chandra S; Fuentes, Gloria

    2014-01-01

    Molecular recognition in biological systems relies on the existence of specific attractive interactions between two partner molecules. Structure-based drug design seeks to identify and optimize such interactions between ligands and their protein targets. The approach followed in medicinal chemistry follows a combination of careful analysis of structural data together with experimental and/or theoretical studies on the system. This chapter focuses on the fact that a protein is not fully characterized by a single structure, but by an ensemble of states, some of them represent "hidden conformations" with cryptic binding sites. We highlight case studies where both experimental and computational methods have been used to mutually drive each other in an attempt to improve the success of the drug design approaches.Advances in both experimental techniques and computational methods have greatly improved our physico-chemical understanding of the functional mechanisms in biomolecules and opened a debate about the interplay between molecular structure and biomolecular function. The beautiful static pictures of protein structures may have led to neglecting the intrinsic protein flexibility, however we are entering a new era where more sophisticated methods are used to exploit this ability of macromolecules, and this will definitely lead to the inclusion of the notion in the pharmaceutical field of drug design.

  8. Competing Discourses about Youth Sexual Exploitation in Canadian News Media

    PubMed Central

    Saewyc, Elizabeth M.; Miller, Bonnie B.; Rivers, Robert; Matthews, Jennifer; Hilario, Carla; Hirakata, Pam

    2015-01-01

    Media holds the power to create, maintain, or break down stigmatizing attitudes, which affect policies, funding, and services. To understand how Canadian news media depicts the commercial sexual exploitation of children and youth, we examined 835 Canadian newspaper articles from 1989–2008 using a mixed methods critical discourse analysis approach, comparing representations to existing research about sexually exploited youth. Despite research evidence that equal rates of boys and girls experience exploitation, Canadian news media depicted exploited youth predominantly as heterosexual girls, and described them alternately as victims or workers in a trade, often both in the same story. News media mentioned exploiters far less often than victims, and portrayed them almost exclusively as male, most often called ‘customers’ or ‘consumers,’ and occasionally ‘predators’; in contrast, research has documented the majority of sexually exploited boys report female exploiters. Few news stories over the past two decades portrayed the diversity of victims, perpetrators, and venues of exploitation reported in research. The focus on victims but not exploiters helps perpetuate stereotypes of sexual exploitation as business or a ‘victimless crime,’ maintains the status quo, and blurs responsibility for protecting youth under the UN Convention on the Rights of the Child. Health care providers and researchers can be advocates for accuracy in media coverage about sexual exploitation; news reporters and editors should focus on exploiters more than victims, draw on existing research evidence to avoid perpetuating stereotypes, and use accurate terms, such as commercial sexual exploitation, rather than terms related to business or trade. PMID:26793015

  9. Aspects of abuse: commercial sexual exploitation of children.

    PubMed

    Jimenez, Melissa; Jackson, Allison M; Deye, Katherine

    2015-03-01

    Commercial sexual exploitation of children (CSEC) and adolescents is a serious worldwide problem. It is, in essence, the sexual abuse of a minor for economic gain. In the United States, there is no uniform nationwide database to capture the incidence and prevalence of CSEC. Therefore, there is a great variation in the estimates, but the actual numbers are unknown. Given the clandestine nature of the practice, it is often underreported and underidentified. Healthcare providers will often encounter victims of commercial sexual exploitation due to mental health, physical health, and sexual health consequences, and therefore should be knowledgeable in the signs of possible sexual exploitation. The aim of this article is to educate healthcare providers on how vulnerable children may become sexually exploited, the health consequences involved with sexual exploitation, how to better identify possible victims, and the medical evaluation of a victim of sexual exploitation. Copyright © 2015 Mosby, Inc. All rights reserved.

  10. Transnational gestational surrogacy: does it have to be exploitative?

    PubMed

    Kirby, Jeffrey

    2014-01-01

    This article explores the controversial practice of transnational gestational surrogacy and poses a provocative question: Does it have to be exploitative? Various existing models of exploitation are considered and a novel exploitation-evaluation heuristic is introduced to assist in the analysis of the potentially exploitative dimensions/elements of complex health-related practices. On the basis of application of the heuristic, I conclude that transnational gestational surrogacy, as currently practiced in low-income country settings (such as rural, western India), is exploitative of surrogate women. Arising out of consideration of the heuristic's exploitation conditions, a set of public education and enabled choice, enhanced protections, and empowerment reforms to transnational gestational surrogacy practice is proposed that, if incorporated into a national regulatory framework and actualized within a low income country, could possibly render such practice nonexploitative.

  11. Proba-V Mission Exploitation Platform

    NASA Astrophysics Data System (ADS)

    Goor, Erwin; Dries, Jeroen

    2017-04-01

    VITO and partners developed the Proba-V Mission Exploitation Platform (MEP) as an end-to-end solution to drastically improve the exploitation of the Proba-V (a Copernicus contributing mission) EO-data archive (http://proba-v.vgt.vito.be/), the past mission SPOT-VEGETATION and derived vegetation parameters by researchers, service providers and end-users. The analysis of time series of data (+1PB) is addressed, as well as the large scale on-demand processing of near real-time data on a powerful and scalable processing environment. Furthermore data from the Copernicus Global Land Service is in scope of the platform. From November 2015 an operational Proba-V MEP environment, as an ESA operation service, is gradually deployed at the VITO data center with direct access to the complete data archive. Since autumn 2016 the platform is operational and yet several applications are released to the users, e.g. - A time series viewer, showing the evolution of Proba-V bands and derived vegetation parameters from the Copernicus Global Land Service for any area of interest. - Full-resolution viewing services for the complete data archive. - On-demand processing chains on a powerfull Hadoop/Spark backend e.g. for the calculation of N-daily composites. - Virtual Machines can be provided with access to the data archive and tools to work with this data, e.g. various toolboxes (GDAL, QGIS, GrassGIS, SNAP toolbox, …) and support for R and Python. This allows users to immediately work with the data without having to install tools or download data, but as well to design, debug and test applications on the platform. - A prototype of jupyter Notebooks is available with some examples worked out to show the potential of the data. Today the platform is used by several third party projects to perform R&D activities on the data, and to develop/host data analysis toolboxes. In parallel the platform is further improved and extended. From the MEP PROBA-V, access to Sentinel-2 and landsat data will

  12. Exploiting Untapped Information Resources in Earth Science

    NASA Astrophysics Data System (ADS)

    Ramachandran, R.; Fox, P. A.; Kempler, S.; Maskey, M.

    2015-12-01

    One of the continuing challenges in any Earth science investigation is the amount of time and effort required for data preparation before analysis can begin. Current Earth science data and information systems have their own shortcomings. For example, the current data search systems are designed with the assumption that researchers find data primarily by metadata searches on instrument or geophysical keywords, assuming that users have sufficient knowledge of the domain vocabulary to be able to effectively utilize the search catalogs. These systems lack support for new or interdisciplinary researchers who may be unfamiliar with the domain vocabulary or the breadth of relevant data available. There is clearly a need to innovate and evolve current data and information systems in order to improve data discovery and exploration capabilities to substantially reduce the data preparation time and effort. We assert that Earth science metadata assets are dark resources, information resources that organizations collect, process, and store for regular business or operational activities but fail to utilize for other purposes. The challenge for any organization is to recognize, identify and effectively utilize the dark data stores in their institutional repositories to better serve their stakeholders. NASA Earth science metadata catalogs contain dark resources consisting of structured information, free form descriptions of data and pre-generated images. With the addition of emerging semantic technologies, such catalogs can be fully utilized beyond their original design intent of supporting current search functionality. In this presentation, we will describe our approach of exploiting these information resources to provide novel data discovery and exploration pathways to science and education communities

  13. Exploitation of Parallelism in Climate Models

    SciTech Connect

    Baer, F.; Tribbia, J.J.; Williamson, D.L.

    1999-03-01

    The US Department of Energy (DOE), through its CHAMMP initiative, hopes to develop the capability to make meaningful regional climate forecasts on time scales exceeding a decade, such capability to be based on numerical prediction type models. We propose research to contribute to each of the specific items enumerated in the CHAMMP announcement (Notice 91-3); i.e., to consider theoretical limits to prediction of climate and climate change on appropriate time scales, to develop new mathematical techniques to utilize massively parallel processors (MPP), to actually utilize MPPs as a research tool, and to develop improved representations of some processes essential to climate prediction. In particular, our goals are to: (1) Reconfigure the prediction equations such that the time iteration process can be compressed by use of MMP architecture, and to develop appropriate algorithms. (2) Develop local subgrid scale models which can provide time and space dependent parameterization for a state- of-the-art climate model to minimize the scale resolution necessary for a climate model, and to utilize MPP capability to simultaneously integrate those subgrid models and their statistics. (3) Capitalize on the MPP architecture to study the inherent ensemble nature of the climate problem. By careful choice of initial states, many realizations of the climate system can be determined concurrently and more realistic assessments of the climate prediction can be made in a realistic time frame. To explore these initiatives, we will exploit all available computing technology, and in particular MPP machines. We anticipate that significant improvements in modeling of climate on the decadal and longer time scales for regional space scales will result from our efforts.

  14. Beyond Fair Benefits: Reconsidering Exploitation Arguments Against Organ Markets.

    PubMed

    Koplin, Julian J

    2017-02-04

    One common objection to establishing regulated live donor organ markets is that such markets would be exploitative. Perhaps surprisingly, exploitation arguments against organ markets have been widely rejected in the philosophical literature on the subject. It is often argued that concerns about exploitation should be addressed by increasing the price paid to organ sellers, not by banning the trade outright. I argue that this analysis rests on a particular conception of exploitation (which I refer to as 'fair benefits' exploitation), and outline two additional ways that the charge of exploitation can be understood (which I discuss in terms of 'fair process' exploitation and complicity in injustice). I argue that while increasing payments to organ sellers may mitigate or eliminate fair benefits exploitation, such measures will not necessarily address fair process exploitation or complicity in injustice. I further argue that each of these three forms of wrongdoing is relevant to the ethics of paid living organ donation, as well as the design of public policy more generally.

  15. Nanostructures Exploit Hybrid-Polariton Resonances

    NASA Technical Reports Server (NTRS)

    Anderson, Mark

    2008-01-01

    Nanostructured devices that exploit the hybrid-polariton resonances arising from coupling among photons, phonons, and plasmons are subjects of research directed toward the development of infrared-spectroscopic sensors for measuring extremely small quantities of molecules of interest. The spectroscopic techniques in question are surface enhanced Raman scattering (SERS) and surface enhanced infrared absorption (SEIRA). An important intermediate goal of this research is to increase the sensitivity achievable by these techniques. The basic idea of the approach being followed in this research is to engineer nanostructured devices and thereby engineer their hybrid-polariton resonances to concentrate infrared radiation incident upon their surfaces in such a manner as to increase the absorption of the radiation for SEIRA and measure the frequency shifts of surface vibrational modes. The underlying hybrid-polariton-resonance concept is best described by reference to experimental devices that have been built and tested to demonstrate the concept. The nanostructure of each such device includes a matrix of silicon carbide particles of approximately 1 micron in diameter that are supported on a potassium bromide (KBr) or poly(tetrafluoroethylene) [PTFE] window. These grains are sputter-coated with gold grains of 40-nm size (see figure). From the perspective of classical electrodynamics, in this nanostructure, that includes a particulate or otherwise rough surface, the electric-field portion of an incident electromagnetic field becomes concentrated on the particles when optical resonance conditions are met. Going beyond the perspective of classical electrodynamics, it can be seen that when the resonance frequencies of surface phonons and surface plasmons overlap, the coupling of the resonances gives rise to an enhanced radiation-absorption or -scattering mechanism. The sizes, shapes, and aggregation of the particles determine the frequencies of the resonances. Hence, the task of

  16. Simulation of subsea gas hydrate exploitation

    NASA Astrophysics Data System (ADS)

    Janicki, Georg; Schlüter, Stefan; Hennig, Torsten; Deerberg, Görge

    2014-05-01

    The recovery of methane from gas hydrate layers that have been detected in several subsea sediments and permafrost regions around the world is a promising perspective to overcome future shortages in natural gas supply. Being aware that conventional natural gas resources are limited, research is going on to develop technologies for the production of natural gas from such new sources. Thus various research programs have started since the early 1990s in Japan, USA, Canada, India, and Germany to investigate hydrate deposits and develop required technologies. In recent years, intensive research has focussed on the capture and storage of CO2 from combustion processes to reduce climate impact. While different natural or man-made reservoirs like deep aquifers, exhausted oil and gas deposits or other geological formations are considered to store gaseous or liquid CO2, the storage of CO2 as hydrate in former methane hydrate fields is another promising alternative. Due to beneficial stability conditions, methane recovery may be well combined with CO2 storage in the form of hydrates. Regarding technological implementation many problems have to be overcome. Especially mixing, heat and mass transfer in the reservoir are limiting factors causing very long process times. Within the scope of the German research project »SUGAR« different technological approaches for the optimized exploitation of gas hydrate deposits are evaluated and compared by means of dynamic system simulations and analysis. Detailed mathematical models for the most relevant chemical and physical processes are developed. The basic mechanisms of gas hydrate formation/dissociation and heat and mass transport in porous media are considered and implemented into simulation programs. Simulations based on geological field data have been carried out. The studies focus on the potential of gas production from turbidites and their fitness for CO2 storage. The effects occurring during gas production and CO2 storage within

  17. Simulated population responses of common carp to commercial exploitation

    SciTech Connect

    Weber, Michael J.; Hennen, Matthew J.; Brown, Michael L.

    2011-12-01

    Common carp Cyprinus carpio is a widespread invasive species that can become highly abundant and impose deleterious ecosystem effects. Thus, aquatic resource managers are interested in controlling common carp populations. Control of invasive common carp populations is difficult, due in part to the inherent uncertainty of how populations respond to exploitation. To understand how common carp populations respond to exploitation, we evaluated common carp population dynamics (recruitment, growth, and mortality) in three natural lakes in eastern South Dakota. Common carp exhibited similar population dynamics across these three systems that were characterized by consistent recruitment (ages 3 to 15 years present), fast growth (K = 0.37 to 0.59), and low mortality (A = 1 to 7%). We then modeled the effects of commercial exploitation on size structure, abundance, and egg production to determine its utility as a management tool to control populations. All three populations responded similarly to exploitation simulations with a 575-mm length restriction, representing commercial gear selectivity. Simulated common carp size structure modestly declined (9 to 37%) in all simulations. Abundance of common carp declined dramatically (28 to 56%) at low levels of exploitation (0 to 20%) but exploitation >40% had little additive effect and populations were only reduced by 49 to 79% despite high exploitation (>90%). Maximum lifetime egg production was reduced from 77 to 89% at a moderate level of exploitation (40%), indicating the potential for recruitment overfishing. Exploitation further reduced common carp size structure, abundance, and egg production when simulations were not size selective. Our results provide insights to how common carp populations may respond to exploitation. Although commercial exploitation may be able to partially control populations, an integrated removal approach that removes all sizes of common carp has a greater chance of controlling population abundance

  18. Exploiting for medical and biological applications

    NASA Astrophysics Data System (ADS)

    Giano, Michael C.

    Biotherapeutics are an emerging class of drug composed of molecules ranging in sizes from peptides to large proteins. Due to their poor stability and mucosal membrane permeability, biotherapeutics are administered by a parenteral method (i.e., syringe, intravenous or intramuscular). Therapeutics delivered systemically often experience short half-lives. While, local administration may involve invasive surgical procedures and suffer from poor retention at the site of application. To compensate, the patient receives frequent doses of highly concentrated therapeutic. Unfortunately, the off-target side effects and discomfort associated with multiple injections results in poor patient compliance. Therefore, new delivery methods which can improve therapeutic retention, reduce the frequency of administration and may aid in decreasing the off-target side effects is a necessity. Hydrogels are a class of biomaterials that are gaining interests for tissue engineering and drug delivery applications. Hydrogel materials are defined as porous, 3-dimensional networks that are primarily composed of water. Generally, they are mechanically rigid, cytocompatible and easily chemically functionalized. Collectively, these properties make hydrogels fantastic candidates to perform as drug delivery depots. Current hydrogel delivery systems physically entrap the target therapeutic which is then subsequently released over time at the site of administration. The swelling and degradation of the material effect the diffusion of the therapy from the hydrogel, and therefore should be controlled. Although these strategies provide some regulation over therapeutic release, full control of the delivery is not achieved. Newer approaches are focused on designing hydrogels that exploit known interactions, covalently attach the therapy or respond to an external stimulus in an effort to gain improved control over the therapy's release. Unfortunately, the biotherapeutic is typically required to be chemically

  19. Early Sexual Exploitation as an Influence in Prostitution.

    ERIC Educational Resources Information Center

    Silbert, Mimi H.; Pines, Ayala M.

    1983-01-01

    Surveyed 200 female street prostitutes to determine whether they were sexually exploited during childhood. Results showed 60 percent of the subjects were sexually exploited. The few girls who discussed their abuse with others were met with shame and most often inaction. Only 10 percent were abused by strangers. (JAC)

  20. A Descriptive Study on Sexually Exploited Children in Residential Treatment

    ERIC Educational Resources Information Center

    Twill, Sarah E.; Green, Denise M.; Traylor, Amy

    2010-01-01

    Sexual exploitation and prostitution of children and adolescents is a multibillion dollar industry in the United States (Estes and Weiner in "Medical, legal & social science aspects of child sexual exploitation: A comprehensive review of pornography, prostitution, and internet crimes, vol I," G.W. Medical Publishing, Inc, St Louis,…