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antibodies to anaplasma marginale major surface proteins 1a and 1b inhibit infectivity for cultured tick cells.major surface protein 1 (msp1) of the cattle pathogen anaplasma marginale (rickettsiales: anaplasmataceae) is a complex of two proteins, msp1a and msp1b. previous studies demonstrated that msp1a and msp1b are adhesins for bovine erythrocytes, while only msp1a proved to be an adhesin for tick cells. in this study, a tick cell culture system for propagation of a. marginale was used to develop an infection inhibition assay for testing the ability of antisera to block infection of a. marginale for c ...200312531299
mechanisms of variable p44 expression by anaplasma phagocytophilum.the human intragranulocytic bacterium anaplasma phagocytophilum promotes variation of p44s, which are surface-exposed proteins encoded by a p44 multigene family. in the present study, the specific p44 gene expression loci in four strains of a. phagocytophilum were identified and it was determined that each consisted of four tandem genes, tr1, omp-1x, omp-1n, and p44. a putative sigma(70)-type promoter was found upstream of tr1. the p44 genes include a central hypervariable region flanked by cons ...200314500485
invasion and survival strategies of anaplasma phagocytophilum.anaplasma phagocytophilum is an aetiological agent of human granulocytic ehrlichiosis, an emerging tick-borne zoonosis in the united states and europe. this obligate intracellular bacterium is unique in that it colonizes polymorphonuclear leucocytes (neutrophils). neutrophils are key players in innate immunity. these short-lived phagocytes ingest invading microorganisms and destroy them by various means, which include fusing the bacteria-containing phagosome with acidic lysosomes as well as dire ...200314531890
obligatory intracellular parasitism by ehrlichia chaffeensis and anaplasma phagocytophilum involves caveolae and glycosylphosphatidylinositol-anchored proteins.obligatory intracellular, human ehrlichiosis agents ehrlichia chaffeensis and anaplasma phagocytophilum create unique replicative compartments devoid of lysosomal markers in monocytes/macrophages and granulocytes respectively. the entry of these bacteria requires host phospholipase c (plc)-gamma2 and protein tyrosine kinases, but their entry route is still unclear. here, using specific inhibitors, double immunofluorescence labelling and the fractionation of lipid rafts, we demonstrate that bacte ...200314531896
predominance of ehrlichia ewingii in missouri dogs.to investigate the species distribution of ehrlichia present in missouri dogs, we tested 78 dogs suspected of having acute ehrlichiosis and 10 healthy dogs. blood from each dog was screened with a broad-range 16s rrna gene pcr assay that detects known pathogenic species of ehrlichia and anaplasma: the species was determined by using species-specific pcr assays and nucleotide sequencing. ehrlichia antibody testing was performed by using an indirect immunofluorescence assay with ehrlichia chaffeen ...200314532192
antigens and alternatives for control of anaplasma marginale infection in cattle.anaplasmosis, a tick-borne cattle disease caused by the rickettsia anaplasma marginale, is endemic in tropical and subtropical areas of the world. the disease causes considerable economic loss to both the dairy and beef industries worldwide. analyses of 16s rrna, groesl, and surface proteins have resulted in the recent reclassification of the order rickettsiales. the genus anaplasma, of which a. marginale is the type species, now also includes a. bovis, a. platys, and a. phagocytophilum, which w ...200314557295
coinfection in patients with lyme disease: how big a risk? 200314557980
molecular methods for ehrlichiosis and lyme disease.this article reviews molecular techniques that have been developed and are effective in the clinical laboratory for the emerging tick-borne infections, ehrlichiosis and lyme disease.200314711096
canine ehrlichiosis.ehrlichia spp. are obligate intracellular bacteria with tropism for hematopoietic cells. monocytic ehrlichioses in dogs and humans are transmitted by ticks and primarily caused by e. canis and e. chaffeensis, respectively. e. canis causes canine monocytic ehrlichioses (cme), a potentially fatal disease in dogs that requires rapid and accurate diagnosis in order to initiate appropriate therapy leading to a favorable prognosis. cme is characterized by three stages; 1) acute, 2) subclinical and 3) ...200314677903
molecular evidence for anaplasma phagocytophilum and borrelia burgdorferi sensu lato in ixodes ricinus ticks from eastern slovakia.ixodes ricinus ticks (20 males, 20 females and 20 nymphs) collected in kosice, slovakia were examined for the presence of anaplasma phagocytophilum and borrelia burgdorferi sensu lato (s.l.) by pcr. 38.3 % of the tested ticks carried single infection of b. burgdorferi s.l. and 8.3 % were infected with a. phagocytophilum. double infection of both pathogens was detected in 5 % of tested ticks. these results indicate that both b. burgdorferi s.l. and a. phagocytophilum co-circulate in the enzootic ...200314677924
detection of borrelia burgdorferi, ehrlichia chaffeensis, and anaplasma phagocytophilum in ticks (acari: ixodidae) from a coastal region of california.a study was conducted in santa cruz county to estimate the prevalence and distribution of the agents of lyme disease, human granulocytic (hge), and human monocytic (hme) ehrlichiosis in 1,187 adult ixodid ticks collected from eight public-use recreation areas over a 2-yr period. borrelia burgdorferi, the causative agent of lyme disease, was detected by a polymerase chain reaction (pcr) in 44 of 776 (5.67%) ixodes pacificus ticks and in 3 of 58 (5.17%) dermacentor variabilis ticks. anaplasma phag ...200314680123
causes of febrile illnesses after a tick bite in slovenian children.to establish the etiology in slovenian children with febrile illnesses occurring after a tick bite.200314688569
ehrlichiosis and related infections. 200314690204
enzootic transmission of babesia divergens among cottontail rabbits on nantucket island, massachusetts.specific ticks seem to locally serve as vector for characteristic microbial assemblages (guilds) comprising spirochetes, piroplasms, ehrlichiae, and arboviruses. borrelia andersoni and anaplasma phagocytophilum are intensely transmitted between cottontail rabbits. to test the hypothesis that a piroplasm may also be maintained in rabbits, we sampled these hosts from nantucket island, massachusetts and tested their blood and tissues by a polymerase chain reaction for evidence of infection. surpris ...200314695079
high serum folate values in lambs experimentally infected with anaplasma phagocytophilum. 200315074632
serological evidence of exposure to tick fever organisms in young cattle on queensland dairy farms.to compare the features of farms on which the exposure of young cattle to tick fever organisms is sufficient to ensure that immunity is high and the risk of clinical disease is low (endemic stability) with those of farms on which exposure is insufficient to induce widespread immunity (hence without endemic stability); to examine the relationships between the management of ticks and tick fever, and endemic stability to babesia bovis, b. bigemina and anaplasma marginale.200315080428
seasonal dynamics of anaplasma phagocytophila in a rodent-tick (ixodes trianguliceps) system, united kingdom.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 activ ...200312533283
conservation of a gene conversion mechanism in two distantly related paralogues of anaplasma marginale.anaplasmataceae, the causative agents of anaplasmosis and ehrlichiosis, persist in the bloodstream of their mammalian hosts, allowing acquisition and transmission by tick vectors. anaplasma marginale establishes persistent infection characterized by sequential cycles of rickettsaemia in which new antigenic variants emerge. the two most immunodominant outer membrane proteins, msp2 and msp3, are paralogues, each encoded by a distinct family of related genes. this study demonstrates that, although ...200312535066
simultaneous infection with borrelia burgdorferi and human granulocytic ehrlichiosis.a 5-year-old child with simultaneous early lyme disease and human granulocytic ehrlichiosis (hge) is described. because of the shared vector, hge and lyme disease are increasingly identified as coinfections in tick-exposed patients. early recognition of concurrent lyme disease and hge is important because amoxicillin, an antibiotic of choice for young children with early lyme disease, is ineffective for hge.200312553302
comparison of pcr assays for detection of the agent of human granulocytic ehrlichiosis, anaplasma phagocytophilum.human granulocytic ehrlichiosis is an emerging infectious disease in the united states and europe, and pcr methods have been shown to be effective for the diagnosis of acute infections. numerous pcr assays and primer sets have been reported in the literature. the analytical sensitivities (limits of detection) of 13 published pcr primer sets were compared using dna extracted from serial dilutions of anaplasma phagocytophilum-infected hl-60 cells. the specificity of the assays that were able to de ...200312574272
co-feeding studies of ticks infected with anaplasma marginale.ticks often cluster at preferred feeding sites on hosts, and the co-feeding of ticks at the same site has been shown to increase feeding success and the transmission of some pathogens. while the major route of infection of ticks with pathogens is via the bloodmeal during feeding on a parasitemic host, non-systemic transmission of viruses and spirochetes has been shown to occur from infected to uninfected ticks at common feeding sites on uninfected hosts. in this research, two separate studies we ...200312623209
detection of ehrlichia spp. in the blood of wild white-tailed deer in missouri by pcr assay and serologic analysis.blood samples collected from wild deer in missouri in november of 2000 and 2001 were positive by pcr assays for ehrlichia chaffeensis (50 of 217; 23%), ehrlichia ewingii (44 of 217; 20%), and anaplasma species (214 of 217; 99%). nucleotide sequences of selected amplicons from the assay for anaplasma matched sequences of the white-tailed deer agent. serologic analysis of 112 deer sampled in 2000 showed a very high prevalence of antibodies to e. chaffeensis (97 of 112; 87%) and a low prevalence of ...200312624063
ehrlichia platys (anaplasma platys) in dogs from maracaibo, venezuela: an ultrastructural study of experimental and natural infections.since 1982 ehrlichia platys, now emended as anaplasma platys, has been diagnosed in dogs from maracaibo, venezuela, using buffy coat smears stained with dip quick. three dogs were inoculated with an a. platys strain. when parasitemia reached 60-97%, blood samples obtained from the inoculated dogs and from two naturally infected dogs were centrifuged to obtain platelet-rich plasma, which was mixed with 0.1% glutaraldehyde at 37 c for 10 minutes. platelet pellets were fixed in 3% glutaraldehyde fo ...200312637754
the hypervariable region of anaplasma marginale major surface protein 2 (msp2) contains multiple immunodominant cd4+ t lymphocyte epitopes that elicit variant-specific proliferative and ifn-gamma responses in msp2 vaccinates.major surface protein 2 (msp2) is an immunodominant outer membrane protein of anaplasma marginale and anaplasma phagocytophilum pathogens that cause bovine anaplasmosis and human granulocytic ehrlichiosis, respectively. msp2 has a central hypervariable region (hvr) flanked by highly conserved amino and carboxyl termini. during a. marginale infection, dynamic and extensive amino acid sequence variation in msp2 occurs through recombination of msp2 pseudogenes into the msp2 expression site, followe ...200312646645
roles of neutrophil beta 2 integrins in kinetics of bacteremia, extravasation, and tick acquisition of anaplasma phagocytophila in mice.tick saliva contains anti-inflammatory and immunosuppressive substances that facilitate blood feeding and enhance tick-vectored pathogen transmission, including anaplasma phagocytophila an etiologic agent of granulocytic ehrlichiosis. as such, inflammation at a tick-feeding site is strikingly different than that typically observed at other sites of inflammation. up-regulation of cd11b/cd18 occurs in host granulocytes following interaction or infection with a phagocytophila, and the absence of cd ...200312480703
rickettsial, ehrlichial and bartonella infections of the myocardium and pericardium.myocarditis and pericarditis are uncommon complications of human rickettsial, ehrlichial and bartonella infections. myocardial inflammation usually occurs in the setting of acute disseminated infection. organisms associated with myocarditis include: rickettsia rickettsii, r. conorii, orientia tsutsugamushi, coxiella burnetii, anaplasma phagocytophila (the causative agent of human granulocytic ehrlichiosis) and bartonella henselae. pericarditis has been described in the setting of r. conorii and ...200312456377
characterization of the functional domain of major surface protein 1a involved in adhesion of the rickettsia anaplasma marginale to host cells.the major surface protein (msp) 1a of the genus type species anaplasma marginale (rickettsiales: anaplasmataceae) has been shown to mediate adhesion, infection and transmission of the organism, as well as to contribute to protective immunity in cattle. msp1a contains a variable number of tandemly repeated peptides in the amino-terminal region, while the remainder of the protein is highly conserved among isolates. the number of repeats varies among geographic isolates of a. marginale but is const ...200312458174
identification of ehrlichia chaffeensis, anaplasma phagocytophilum, and a. bovis in haemaphysalis longicornis and ixodes persulcatus ticks from korea.a total of 1,467 tick (1,463 of haemaphysalis longicornis, three of ixodes persulcatus and one of i. turdus) collected from nine provinces of korea were examined by taqman real-time pcr for the presence of ehrlichia and anaplasma species. one set of primers and a probe were designed for detection of all of the ehrlichia and anaplasma species. template dnas (total 803) were prepared either from pools of larvae, nymphs, adult males and females, or from the salivary gland and midgut of adult ticks. ...200312804377
cpg odn 2006 and il-12 are comparable for priming th1 lymphocyte and igg responses in cattle immunized with a rickettsial outer membrane protein in alum.immunostimulatory oligodeoxynucleotides containing unmethylated cpg dinucleotides (cpg odn) stimulate il-12-dependent th1 dominated cytokine and enhanced igg responses when co-delivered with antigen to mice. however, the cpg odn sequences that are optimal for each mammalian species may differ. previously, we demonstrated that a cpg odn containing the gtcgtt motif was optimal for stimulating bovine b cell proliferation, and induced il-6, il-12 and ifn-gamma production by peripheral blood mononucl ...200312804862
prevalence of anaplasma phagocytophila and borrelia burgdorferi in ixodes persulcatus ticks from northeastern china.a total of 1,345 ixodes persulcatus ticks collected from northeastern china were investigated for the presence of anaplasma phagocytophila and borrelia burgdorferi by a nested polymerase chain reaction (pcr). sixty-two (4.6%) ticks were positive for a. phagocytophila and 454 (33.8%) were positive for b. burgdorferi. seven (0.5%) were coinfected with both agents. sequence analysis of 919-basepair pcr amplicons revealed three types of a. phagocytophila. type 1 was identical to the published sequen ...200312812342
major surface protein 2 of anaplasma phagocytophilum facilitates adherence to granulocytes.anaplasma phagocytophilum is an obligate intracellular bacterium that infects myeloid cells in the mammalian host. msp2 (p44) is the major immunodominant outer-membrane protein of these bacteria. we hypothesized that msp2 acts as an adhesin for a. phagocytophilum entry into granulocytes. this potential role was investigated by blocking binding with msp2 monoclonal antibodies and by antagonizing binding and propagation with recombinant msp2 (rmsp2) in vitro. with hl-60 cells, fresh human peripher ...200312819090
cloning and characterization of putative zinc protease genes of ehrlichia canis.a putative zinc protease gene operon from ehrlichia canis was cloned and sequenced. a genomic library was constructed in a phg165 plasmid vector using sau3a partially digested e. canis chromosomal dna. sequence analysis of the insert dna from this clone indicated two open reading frames with a size of 1314 and 1350 bp that encodes for proa and prob, respectively. based on blast analyses, proa and prob share 20-30% identities with members of the eukaryotic mitochondrial processing peptidase (mpp) ...200312825352
humoral immune response and hematologic evaluation of pregnant jersey cows after vaccination with anaplasma centrale.the main objective of this work was to evaluate the safety of an anaplasma centrale vaccine in pregnant pure bred jersey cows selected from a herd located at miranda state, venezuela. ten cows of 3-5 months of gestation were chosen and previous vaccination all cows were tested for anaplasma antibodies by the indirect immunofluorescence assay (ifa), so only seronegative cows were included in the group, and for blood parameters, rectal temperature, and pregnancy. selected cows were vaccinated intr ...200312829387
purification of ovine neutrophils and eosinophils: anaplasma phagocytophilum affects neutrophil density.studies on the functions of ovine granulocytes require pure and functionally active populations of neutrophils and eosinophils. this report describes an improved technique for the separation of neutrophils and eosinophils from the peripheral blood of sheep infected with anaplasma phagocytophilum and from normal sheep. after centrifugation and discarding the buffy coat layer, which contains the bulk of mononuclear cells, neutrophils with a high degree of purity (94.87 [+/-1.7]%, n=9) and good yie ...200312834611
analysis of genetic identity of north american anaplasma phagocytophilum strains by pulsed-field gel electrophoresis.biological and geographic heterogeneity of anthropozoonosis caused by anaplasma phagocytophilum is poorly understood. seven north american a. phagocytophilum strains were compared by pfge. the average genome size was 1.58 mbp, and restriction patterns were identical. new world strains of a. phagocytophilum have a large genome and a high degree of genetic uniformity.200312843101
structurally distinct requirements for binding of p-selectin glycoprotein ligand-1 and sialyl lewis x to anaplasma phagocytophilum and p-selectin.colonization of neutrophils by the bacterium anaplasma phagocytophilum causes the disease human granulocytic ehrlichiosis. the pathogen also infects mice, its natural host. like binding of p-selectin, binding of a. phagocytophilum to human neutrophils requires expression of p-selectin glycoprotein ligand-1 (psgl-1) and alpha1-3-fucosyltransferases that construct the glycan determinant sialyl lewis x (slex). binding of a. phagocytophilum to murine neutrophils, however, requires expression of alph ...200312847092
differences in clinical manifestations and hematological and serological responses after experimental infection with genetic variants of anaplasma phagocytophilum in sheep.five-month-old lambs were experimentally infected with two 16s rrna genetic variants of anaplasma phagocytophilum, variants 1 (genbank accession no. m73220) and 2 (genbank accession no. af336220). additional sequencing of the groesl heat shock operon gene indicated that these variants differ in three nucleotides at positions 782, 824, and 890. the variants were obtained by blood sampling of a. phagocytophilum-infected lambs from one sheep flock in norway and were stored at -70 degrees c with 10% ...200312853406
ehrlichiae and ehrlichial diseases in china.the various ticks collected from different areas of china were examined for the existence of ehrlichial agents by polymerase chain reaction (pcr) with genus- or species-specific primers designed on the basis of ehrlichial 16s rrna genes and sequence analyses. in southern china, e. chaffeensis was detected in amblyomma testudinarium ticks from infested cattle, haemaphysalis yeni ticks from hare, and ixodes ovatus ticks from muntiacus reevesi. e. canis was identified in rhipicephalus sanguineus ti ...200312860598
transmission efficiency of the ap-variant 1 strain of anaplasma phagocytophila.nymphal ixodes scapularis ticks were collected from several sites in rhode island. dna was extracted from a subset of these ticks, and pcr and dna sequencing of the 16s rrna gene were used to determine the ratio of anaplasma phagocytophila-human agent (ap-ha) to a genetic variant not associated with human disease (ap-variant 1). the remaining ticks were allowed to feed to repletion on either white-footed (peromyscus leucopus) or dba/2 (mus musculus) mice. the engorged ticks, and blood samples dr ...200312860603
investigation of anaplasma phagocytophila infections in ixodes ricinus and dermacentor reticulatus ticks in austria. 200312860606
a survey on anaplasma phagocytophila in wild small mammals and roe deer (capreolus capreolus) in northern spain. 200312860607
infections of wild animals with anaplasma phagocytophila in austria and the czech republic. 200312860608
domestic animals as indicators of anaplasma species infections in northern albania. 200312860610
molecular epidemiological study for tick-borne disease (ehrlichia and anaplasma spp.) surveillance at selected u.s. military training sites/installations in korea.vector-borne diseases are a potential public health threat to u.s. forces korea (usfk). ehrlichia and anaplasma spp., transmitted by ticks, are only two of several diseases that may affect military readiness and operations. rodents were collected at selected u.s. military installations and training sites in the republic of korea. dna was extracted from spleen tissues and assayed by pcr methods for ehrlichia and anaplasma species. from rodents and mustelids collected during 1999 and 2000, a total ...200312860612
characterization of anaplasma phagocytophila and borrelia burgdorferi genotypes in ixodes scapularis ticks from pennsylvania. 200312860614
genetic diversity of bacterial agents detected in ticks removed from asymptomatic patients in northeastern italy.a total of 360 ticks were removed from 353 asymptomatic subjects in belluno province, italy and surrounding areas, from 1998 to 2001. ticks were identified as ixodes ricinus (357), ixodes hexagonus (1), rhipicephalus sanguineus (1), and ixodes ventalloi (1). tick dna was investigated by pcr and subsequent sequencing of amplified products to identity associated bacterial agents. primers targeting different genes of rickettsia (glta and ompa), borrelia (16s rdna, rpob), francisella (16s rdna), and ...200312860623
human granulocytic ehrlichiosis in slovenia.several reports indicate the presence of infection with anaplasma phagocytophila in europe. the first case with confirmed human granulocytic ehrlichiosis was reported from slovenia in 1997. by 2002, 13 patients have been discovered in a prospective study on the etiology of febrile illnesses occurring within six weeks following a tick bite, conducted at the department of infectious diseases, university medical centre, ljubljana, slovenia. the diagnosis was established by molecular identification ...200312860640
comparison of patients fulfilling criteria for confirmed and probable human granulocytic ehrlichiosis. 200312860648
etiology of tick-borne febrile illnesses in slovenian children. 200312860651
multiplex taqman pcr assay for rapid detection of anaplasma phagocytophila and borrelia burgdorferi. 200312860656
clinical and laboratory features of the first detected cases of a. phagocytophila infections in dogs from slovenia. 200312860668
anaplasma phagocytophila as an abortifacient agent in sheep farms from northern spain. 200312860669
fatal cases of tick-borne fever (tbf) in sheep caused by several 16s rrna gene variants of anaplasma phagocytophilum. 200312860670
early induction and late abrogation of respiratory burst in a. phagocytophilum-infected neutrophils.human granulocytic ehrlichiosis is an emerging tick-borne disease caused by the obligate intracellular pathogen, anaplasma phagocytophilum. this organism is unique because it survives and propagates in neutrophil vacuoles. during phagocytosis professional phagocytes increase oxygen consumption (respiratory burst) through the activity of nadph-oxidase, which generates superoxide anion (o2(-)) and hydrogen peroxide (h(2)o(2)). we assayed the ability of a. phagocytophilum-infected neutrophils to ge ...200312860678
route of transmission alters the infectivity of anaplasma phagocytophila in mice. 200312860679
mechanisms to create a safe haven by members of the family anaplasmataceae.members of the family anaplasmataceae are obligatory intracellular bacteria with unique host cell specificities. depending on each bacterial species, granulocytes, platelets, endothelial cells, monocytes, macrophages, red blood cells, and cells of invertebrates are specifically infected. this unique host cell specificity has been the major hurdle to overcome in order to cultivate this group of bacteria. because these bacteria cannot survive outside host cells, once released from a host cell, the ...200312860688
analysis of 16s rrna gene sequences of ehrlichia canis, anaplasma platys, and wolbachia species from canine blood in japan.in the present study, three canine blood samples from japan, that were suspected to be ehrlichia positive were examined. after sequencing the 16s rrna genes, each dog was found to be infected either with ehrlichia canis (kagoshima 1), anaplasma platys (okinawa 1), or wolbachia sp. (okinawa 2). phylogenic analysis was performed on these sequences. the nearly entire 16s rrna sequence of kagoshima 1 was found to be most similar to the sequences from oklahoma and venezuela e. canis strains (1 base p ...200312860708
determination of white-tailed deer agent groesl operon sequences for phylogenetic and diagnostic applications. 200312860709
murine neutrophils require alpha1,3-fucosylation but not psgl-1 for productive infection with anaplasma phagocytophilum.anaplasma phagocytophilum causes human granulocytic ehrlichiosis, the second most common tick-borne disease in the united states. mice are natural reservoirs for this bacterium and man is an inadvertent host. a phagocytophilum's tropism for human neutrophils is linked to neutrophil expression of p-selectin glycoprotein ligand-1 (psgl-1), as well as sialylated and alpha1,3-fucosylated glycans. to determine whether a phagocytophilum uses similar molecular features to infect murine neutrophils, we ...200312869507
diminished adhesion of anaplasma phagocytophilum-infected neutrophils to endothelial cells is associated with reduced expression of leukocyte surface selectin.anaplasma phagocytophilum propagates within neutrophils and causes a disease marked by inflammatory tissue injury or complicated by opportunistic infections. we hypothesized that infection with a. phagocytophilum modifies the binding of neutrophils to endothelial cells and the expression of neutrophil adhesion molecules and studied these changes in vitro. infected dimethyl sulfoxide-differentiated hl-60 cells and neutrophils showed reduced binding to cultured brain and systemic endothelial cells ...200312874338
antigenic characterization of brazilian isolates of anaplasma marginale.antigenic characterization of anaplasma marginale isolates, by identifying conserved and variable epitopes of major surface proteins (msp), is an important tool for vaccine development against this rickettsia. the b cell epitopes of a. marginale isolates from three microregions of the state of pernambuco and one from the state of mato grosso do sul, brazil, were characterized by indirect fluorescent antibody technique (ifat) and western blot (wb) with 15 monoclonal antibodies (mabs). the epitope ...200312886423
enzootic transmission of the agent of human granulocytic ehrlichiosis among cottontail rabbits.to determine whether the agent of human granulocytic ehrlichiosis (hge) (anaplasma phagocytophilum) may be maintained in a parallel enzootic cycle between cottontail rabbits and their ticks, we sampled these hosts from a zoonotic site during five transmission seasons. evidence of infection was sought by microscopy and polymerase chain reaction from rabbit blood or splenic tissue, and from ticks collected from rabbits or from vegetation. approximately 27% of all rabbits sampled contained evidence ...200312887018
characterization of anaplasma marginale isolated from north american bison.anaplasma marginale (rickettsiales: anaplasmataceae), a tick-borne pathogen of cattle, is endemic in tropical and subtropical regions of the world. although serologic tests have identified american bison, bison bison, as being infected with a. marginale, the present study was undertaken to confirm a. marginale infection and to characterize isolates obtained from naturally infected bison in the united states and canada. major surface protein (msp1a and msp4) sequences of bison isolates were chara ...200312902301
enzootic transmission of anaplasma bovis in nantucket cottontail rabbits.serological studies of cottontail rabbits sampled from nantucket island, mass., have suggested exposure to at least two ehrlichiae. the agent of human granulocytic ehrlichiosis (anaplasma phagocytophilum) is intensely enzootic in rabbits there, but the identity of the other ehrlichial infection remains undescribed. we sampled rabbits over five transmission seasons and tested their blood and tissues for evidence of infection using pcr targeting an ehrlichia genus-wide 16s rdna target. sequence an ...200312904385
transmission of anaplasma marginale by boophilus microplus: retention of vector competence in the absence of vector-pathogen interaction.whether arthropod vectors retain competence for transmission of infectious agents in the long-term absence of vector-pathogen interaction is unknown. we addressed this question by quantifying the vector competence of two tick vectors, with mutually exclusive tropical- versus temperate-region distributions, for genetically distinct tropical- and temperate-region strains of the cattle pathogen anaplasma marginale. the tropical cattle tick boophilus microplus, which has been eradicated from the con ...200312904396
importance of primer specificity for pcr detection of anaplasma phagocytophila among ixodes scapularis ticks from wisconsin. 200312904445
modulation of leukocyte populations and immune responses in sheep experimentally infected with anaplasma (formerly ehrlichia) phagocytophilum.anaplasma phagocytophilum infection in sheep is characterized by an immune suppression as indicated by impaired antibody response, reduced lymphocyte response and reduced oxidative burst. the effect of a. phagocytophilum infection on leucocyte populations, especially lymphocytes, was therefore investigated in six sheep experimentally infected with a. phagocytophilum, and compared with leucocyte populations from control animals.to investigate the ability of the infection to interfere with the cel ...200312909412
ehrlichiosis and related infections.ehrlichiosis is a term that has been used to describe infection with any of a number of related intracellular, vector-borne pathogens. a recent reclassification has resulted in the transfer of several species previously known as ehrlichia to the genus anaplasma or neorickettsia. ehrlichia and anaplasma are transmitted largely through the bite of infected ticks, while vectors for neorickettsia include trematodes and the intermediate hosts (i.e., fish, snails, and insects) involved in the trematod ...200312910747
the immunologic response of dogs to bartonella vinsonii subspecies berkhoffii antigens: as assessed by western immunoblot analysis.bartonella vinsonii subspecies berkhoffii is a recently recognized zoonotic pathogen that causes endocarditis, granulomatous rhinitis, and granulomatous lymphadenitis in dogs. isolation of b. vinsonii (berkhoffii) from blood or tissue samples is frequently unsuccessful; therefore, diagnosis is primarily dependent on serologic or molecular testing modalities. because previous canine serologic studies have used an indirect immunofluorescence assay (ifa), without western immunoblot (wi) confirmatio ...200312918816
ehrlichia chaffeensis and anaplasma phagocytophilum lack genes for lipid a biosynthesis and incorporate cholesterol for their survival.ehrlichia chaffeensis and anaplasma phagocytophilum are agents of human monocytic and granulocytic ehrlichioses, respectively. they are extremely sensitive to mechanical stress and are pleomorphic gram-negative bacteria. membrane incorporation of cholesterol from the eukaryotic host is known to be essential for other fragile and pleomorphic bacteria and mycoplasmas that lack a cell wall. thus, we tested whether cholesterol is required for e. chaffeensis and a. phagocytophilum. using a freeze fra ...200312933880
overcoming constraints to meeting increased demand for babesia bigemina vaccine in australia.demand for live trivalent tick fever vaccine containing babesia bovis, babesia bigemina and anaplasma centrale produced by the department of primary industries, queensland, has increased from less than 10,000 doses in 1988 to 500,000 doses in 2001. this paper describes a series of trials aimed at overcoming certain constraints to obtain b. bigemina parasitised erythrocytes (pes) on a large enough scale from infected splenectomised calves to meet the demand. passage through a series of splenectom ...200312935736
serological evidence for human granulocytic ehrlichiosis in western austria.in order to evaluate the seroprevalence of human granulocytic ehrlichiosis in western austria, sera from 357 tyrolean blood donors were tested by an immunofluorescence assay. to assess the concomitant seroreactivity against borrelia burgdorferi sensu lato, sera were further investigated by enzyme-linked immunoassay and western blot. thirty-two sera (9.0%) showed antibodies to anaplasma phagocytophilum at a titre of 1:128 or higher, and 30 (8.4%) were seroreactive against borrelia burgdorferi sen ...200312938008
identification of borrelia burgdorferi sensu lato, anaplasma and ehrlichia species, and spotted fever group rickettsiae in ticks from southeastern europe.prevalence data for tick-borne pathogens are used to assess the risk for human health. in this study the presence and identity of borrelia burgdorferi sensu lato, ehrlichia, anaplasma, and rickettsia species in bulgarian ixodes ricinus ticks and in non- ixodes ticks from turkey and albania was determined by polymerase chain reaction (pcr) and reverse line blot hybridization. in the adult bulgarian ticks, the prevalence of borrelia burgdorferi sensu lato infection was approximately 40%, while bor ...200312938010
epidemiological survey of anaplasma platys and ehrlichia canis using ticks collected from dogs in japan.detection of anaplasma platys and ehrlichia canis in ticks recovered from dogs in japan was attempted using a species-specific nested pcr based on the 16s rrna gene. a total of 1211 ticks recovered from 1211 dogs from all over japan were examined for a. platys and e. canis. four tick samples from fukushima, miyazaki and kagoshima prefectures recovered from four different dogs showed a positive reaction for a. platys. although the four dogs did not show any clinical signs and no blood examination ...200312944048
successful treatment of human granulocytic ehrlichiosis in children using rifampin.human granulocytic ehrlichiosis (hge) is an emerging tick-borne infectious disease caused by anaplasma phagocytophilum. clinical features include a flu-like illness that usually resolves within 1 week. more serious infection may occur that requires hospital admission or culminates in death. doxycycline is the treatment of choice for hge but may cause permanent staining of teeth in children younger than 8 years of age. we report successful treatment of hge with rifampin in 2 children, 4 and 6 yea ...200312949322
human anaplasmosis presenting as atypical pneumonitis in france.human anaplasmosis is a febrile illness caused by anaplasma phagocytophilum, an intracellular bacterium transmitted by ixodes ticks in the united states and europe. although cough is reported in 30% of the american cases, interstitial pneumonitis has been noted only once. of the 9 confirmed cases reported in europe, 3 presented with atypical pneumonitis. a. phagocytophilum should be added to the list of agents responsible for interstitial pneumonitis, especially in areas where human anaplasmosis ...200312955649
isolation of an anaplasma sp. organism from white-tailed deer by tick cell culture.we used tick cell culture to isolate a bacterium previously referred to as the "white-tailed deer (wtd) agent" from two captive fawns inoculated with blood from wild wtd (odocoileus virginianus). buffy coat cells were added to ise6 tick cell cultures and incubated at 34 degrees c, and 8 days later, anaplasma-like inclusions were demonstrated in giemsa-stained culture samples. the microbes became established and could be continuously passaged in tick cells. the identity of a culture isolate desig ...200312958265
development of a polyclonal competitive enzyme-linked immunosorbent assay for detection of antibodies to ehrlichia ruminantium.a polyclonal competitive enzyme-linked immunosorbent assay (pc-elisa) is described for detection of antibodies to ehrlichia (cowdria) ruminantium by using a soluble extract of endothelial cell culture-derived e. ruminantium as the antigen and biotin-labeled polyclonal goat immunoglobulins as the competitor. for goats, the diagnostic sensitivity and specificity were both 100% with a cutoff of 80% inhibition (80 pi), with detection of antibodies for 550 days postinfection. for cattle, diagnostic s ...200312965926
serotyping isolates of anaplasma phagocytophilum by using monoclonal antibodies.ten mouse monoclonal antibodies (mabs) that react with anaplasma phagocytophilum (the human granulocytic ehrlichiosis agent) webster isolates were developed. seven different isolates of a. phagocytophilum were subtyped with these mabs. western blot analysis revealed that these mabs reacted mainly with 41- to 46-kda msp2 proteins. six mabs reacted with all isolates. four other mabs reacted with human isolates from wisconsin, but not with human isolates from new york or with animal isolates. three ...200312965936
identification of novel protective antigens from anaplasma marginale.a successful recombinant vaccine against anaplasma marginale remains elusive, despite intensive study of the protective, though variable major surface antigens. as an alternative approach to the discovery of additional antigens, crude parasite material was subjected to conventional protein fractionation, coupled with vaccination and parasite challenge trials, without making assumptions as to the nature or location of these antigens. this has lead to the identification of four novel antigens that ...200312706672
rna polymerase beta-subunit-based phylogeny of ehrlichia spp., anaplasma spp., neorickettsia spp. and wolbachia pipientis.sequence analysis of rpob, the gene encoding the beta-subunit of rna polymerase, was used in a phylogenetic investigation of nine species from the genera ehrlichia, neorickettsia, wolbachia and anaplasma. the complete nucleotide sequences obtained for anaplasma phagocytophilum (hge agent), ehrlichia chaffeensis, neorickettsia sennetsu, neorickettsia risticii, anaplasma marginale and wolbachia pipientis were amongst the longest rpob sequences in genbank and ranged from 4074 bp for n. sennetsu to ...200312710612
[fever after a tick bite: clinical manifestations and diagnosis of acute tick bite-associated infections in northeastern switzerland].different tick-borne infections can cause an acute febrile illness. the study objectives were to investigate the clinical manifestations and diagnosis of infections among patients who presented with fever after a tick-bite, and to detect newly described pathogens, including ehrlichia, babesia and rickettsia helvetica, in north-eastern switzerland.200312736854
molecular characterization of anaplasma phagocytophilum and borrelia burgdorferi in ixodes scapularis ticks from pennsylvania.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 ...200312682147
detection of ehrlichia spp., anaplasma spp., rickettsia spp., and other eubacteria in ticks from the thai-myanmar border and vietnam.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 aur ...200312682151
anaplasma marginale msp1alpha genotypes evolved under positive selection pressure but are not markers for geographic isolates.anaplasma marginale (order rickettsiales, family anaplasmataceae), a tick-borne pathogen of cattle, is endemic in tropical and subtropical regions of the world. many geographic isolates of a. marginale occur in the united states and have been identified by major surface protein 1a (msp1a), which varies in sequence and molecular weight due to different numbers of tandem 28- to 29-amino-acid repeats. the present study was undertaken to examine the genetic variations among isolates of a. marginale ...200312682152
prospective study of coinfection in patients with erythema migrans.the frequency of coinfection with borrelia burgdorferi and either anaplasma phagocytophila or babesia microti among patients with erythema migrans, the initial skin lesion of lyme disease, was assessed in 2 mainland locations in rhode island and connecticut in a 4-year prospective study. of the 93 patients with culture-proven erythema migrans, 2 (2%) patients had coinfection with a. phagocytophila and 2 (2%) had coinfection with b. microti. we concluded that the frequency of coinfection with the ...200312684924
antibiotic susceptibilities of anaplasma (ehrlichia) phagocytophilum strains from various geographic areas in the united states.we tested the antibiotic susceptibilities of eight strains of anaplasma phagocytophilum (the agent of human granulocytic ehrlichiosis) collected in various geographic areas of the united states, including minnesota, wisconsin, california, and new york. the results are homogeneous and show that doxycycline, rifampin, and levofloxacin are the most active antibiotics against these strains in vitro.200312499227
phylogenetic analysis of the erythrocytic anaplasma species based on 16s rdna and groel (hsp60) sequences of a. marginale, a. centrale, and a. ovis and the specific detection of a. centrale vaccine strain.phenotypic criteria for the identification of erythrocytic ruminant anaplasma species has relied on subjective identification methods such as host pathogenicity (virulence for cattle or sheep) and/or the location of anaplasma inclusion bodies within the host's red cells. sequence comparisons of new and available genbank accessions were investigated to elucidate the relationships among these closely related anaplasma species. twenty-one 16s rdna and groel (hsp60) sequences from 13 anaplasma margi ...200312488078
evidence for modulated immune response to anaplasma phagocytophila sensu lato in cats with fiv-induced immunosuppression.human granulocytic ehrlichiosis (hge) is an emerging infectious disease in which some patients experience unusual opportunistic infections. in this study, cats infected with the hge agent, anaplasma phagocytophila s.l., had clinical granulocytic ehrlichiosis (ge), anti-nuclear antibodies and increased ifn-gamma mrna. in fiv-immunosuppressed cats with ge, there was upregulated il-10 transcription but not ifn-gamma. cats with fiv had poor response to vaccines, regardless of ge status. this prelimi ...200312493491
expression of multiple outer membrane protein sequence variants from a single genomic locus of anaplasma phagocytophilum.anaplasma phagocytophilum is the causative agent of an emerging tick-borne zoonosis in the united states and europe. the organism causes a febrile illness accompanied by other nonspecific symptoms and can be fatal, especially if treatment is delayed. persistence of a. phagocytophilum within mammalian reservoir hosts is important for ensuring continued disease transmission. in the related organism anaplasma marginale, persistence is associated with antigenic variation of the immunoprotective oute ...200312654783
anaplasma phagocytophilum reduces neutrophil apoptosis in vivo.ovine neutrophils spontaneously underwent apoptosis during culture in vitro, as assessed by morphological changes and exposure of annexin v binding sites on their cell surfaces. the addition of conditioned medium from concanavalin a-treated ovine peripheral blood mononuclear cells (pbmc) could partially protect against this progression into apoptosis, but dexamethasone and sodium butyrate could not. actinomycin d accelerated the rate at which ovine neutrophils underwent apoptosis. neutrophils is ...200312654818
transmission of anaplasma phagocytophilum to ixodes ricinus ticks from sheep in the acute and post-acute phases of infection.a total of 60 sheep were exposed to anaplasma phagocytophilum infection on an enclosed area of ixodes ricinus-infested pasture in north wales, united kingdom, and rapidly acquired acute a. phagocytophilum infections detectable by pcr and blood smear examination. of the ticks that had engorged in the previous instar on infected sheep, 52% of adult ticks and 28% of nymphs were pcr positive; a significant, 10-fold increase in prevalence compared to that of ticks that engorged on sheep preinfection ...200312654828
molecular evidence of coinfection of borrelia burgdorferi sensu lato, human granulocytic ehrlichiosis agent, and babesia microti in ticks from northwestern poland.to assess the potential risk for tick-borne agents, ixodes ricinus were collected from 2 sites in northwestern poland. the ticks were tested by polymerase chain reaction for coinfection with borrelia burgdorferi sensu lato (s. l.), human granulocytic ehrlichiosis (hge) agent, and babesia microti. of the 533 processed ticks, 16.7% were positive for b. burgdorferi s. l., 13.3% for b. microti, and 4.5% for the hge agent. twenty ticks were coinfected with 2 or 3 of the pathogens.200312659331
[prevalence of anaplasma marginale and specific antibodies in new born calves].anaplasma marginale is an intraerythrocytic rickettsia that affects the cattle and constitute a constraint for its production. the transplacental transmission of a. marginale could be of importance under field conditions, but until now no data is available to interpret its relevance in venezuela. the goal of this work was to evaluate both the presence of a. marginale and the antibody response against the rickettsia in new born calves. fourteen calves, one week old, were simultaneously bled for p ...200314976783
survey of ticks collected in mississippi for rickettsia, ehrlichia, and borrelia species.from november 1999 through october 2000, we tested ticks collected from vegetation as well as from deer, dogs, and humans for spotted fever group (sfg) rickettsiae, ehrlichia chaffeensis, and borrelia spp. spirochetes. a total of 149 adult ticks representing four species was collected from 11 collection sites from southwestern to northern mississippi. amblyomma americanum was most commonly collected (n=68), followed by ixodes scapularis (n=53). the bird tick, ixodes brunneus (usually rare), was ...200314714667
acarological risk of exposure to agents of tick-borne zoonoses in the first recognized italian focus of lyme borreliosis.acarological risk was calculated as the probability of encountering at least one host-seeking ixodes ricinus tick infected by the pathogen borrelia burgdorferi sensu lato, in 100 m transects in the province of genoa, italy. the seasonal pattern of i. ricinus was studied using generalized estimating equations (gee) with negative binomial error, to consider overdispersion of tick counts and repeated sampling of the same dragging sites from april 1998 to march 1999. prevalence of infection by b. bu ...200314959782
expression cloning.expression cloning involves the selection of specific polypeptides, generated from a cdna or genomic dna library, based on certain characteristics of the expressed proteins, such as antibody or ligand binding, recognition by t-cells, function, or complementation of cell defects. here we describe the detailed construction of a genomic, random shear lambda expression library, adsorption of anti escherichia coli antibody from antiserum, the screening of an expression library with specific antisera, ...200414959824
transmission route efficacy and kinetics of anaplasma phagocytophilum infection in white-footed mouse, peromyscus leucopus.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 tic ...200415682514
sequence analysis of p44 homologs expressed by anaplasma phagocytophilum in infected ticks feeding on naive hosts and in mice infected by tick attachment.the 44-kda immunodominant outer membrane proteins (p44 proteins) of anaplasma phagocytophilum are encoded by the p44 polymorphic multigene family. the present study examined p44 expression and analyzed the cdna sequences of various p44 transcripts from the spleens and blood of mice infected by the bites of ticks infected with the a. phagocytophilum ntn-1 strain or of naturally infected nymphal ticks and in the salivary glands and midgut tissues of these ticks. a total of 300 p44 cdnas were subje ...200414742506
disruption of ixodes scapularis anticoagulation by using rna interference.ixodes scapularis ticks transmit many pathogens, including borrelia burgdorferi, anaplasma phagocytophilum, and babesia microti. vaccines directed against arthropod proteins injected into the host during tick engorgement could prevent numerous infectious diseases. salp14, a salivary anticoagulant, poses a key target for such intervention. salp14 is the prototypic member of a family of potential i. scapularis anticoagulants, expressed and secreted in tick saliva during tick feeding. rna interfere ...200414745044
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