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novel anaplasma and ehrlichia organisms infecting the wildlife of two regions of the brazilian amazon.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. anaplasmat ...201728690147
prevalence of babesia spp., ehrlichia spp., and tick infestations in oklahoma black bears (ursus americanus).american black bears (ursus americanus) are commonly infested with ticks throughout their range, but there are few surveys for tick-borne disease agents in bears. to characterize tick infestations and determine the prevalence of current infection with babesia spp. and past or current infection with ehrlichia spp. in newly re-established populations of black bears in east central and southeastern oklahoma, we identified adult (n=1,048) and immature (n=107) ticks recovered from bears (n=62). we ev ...201728657860
molecular detection of tick-borne pathogen diversities in ticks from livestock and reptiles along the shores and adjacent islands of lake victoria and lake baringo, kenya.although diverse tick-borne pathogens (tbps) are endemic to east africa, with recognized impact on human and livestock health, their diversity and specific interactions with tick and vertebrate host species remain poorly understood in the region. in particular, the role of reptiles in tbp epidemiology remains unknown, despite having been implicated with tbps of livestock among exported tortoises and lizards. understanding tbp ecologies, and the potential role of common reptiles, is critical for ...201728620610
parasites of domestic and wild animals in south africa. l. ixodid ticks infesting horses and donkeys.the aim of the study was to determine the species spectrum of ixodid ticks that infest horses and donkeys in south africa and to identify those species that act as vectors of disease to domestic livestock. ticks were collected opportunistically from 391 horses countrywide by their owners or grooms, or by veterinary students and staff at the faculty of veterinary science, university of pretoria. ticks were also collected from 76 donkeys in limpopo province, 2 in gauteng province and 1 in north we ...201728281774
genetic diversity of <i>ehrlichia ruminantium</i> strains in cameroon.in order to investigate the extent of genetic diversity among ehrlichia ruminantium strains in cameroon, a partial fragment (800 bp) of the e. ruminantium map1 gene was amplified by nested polymerase chain reaction in 121 of 156 e. ruminantium pcs20-positive dna samples extracted from ticks and cattle collected from two ranches. deoxyribonucleic acid sequencing of the map1 gene products indicated the presence of at least 21 genotypes at the nucleotide level and 16 genotypes at the amino acid lev ...201428235281
a comparison of dna extraction protocols from blood spotted on fta cards for the detection of tick-borne pathogens by reverse line blot hybridization.an essential step in the molecular detection of tick-borne pathogens (tbps) in blood is the extraction of dna. when cooled storage of blood under field conditions prior to dna extraction in a dedicated laboratory is not possible, the storage of blood on filter paper forms a promising alternative. we evaluated six dna extraction methods from blood spotted on fta classic(®) cards (fta cards), to determine the optimal protocol for the subsequent molecular detection of tbps by pcr and the reverse li ...201727825733
recombination is a major driving force of genetic diversity in the anaplasmataceae ehrlichia ruminantium.the disease, heartwater, caused by the anaplasmataceae e. ruminantium, represents a major problem for tropical livestock and wild ruminants. up to now, no effective vaccine has been available due to a limited cross protection of vaccinal strains on field strains and a high genetic diversity of ehrlichia ruminantium within geographical locations. to address this issue, we inferred the genetic diversity and population structure of 194 e. ruminantium isolates circulating worldwide using multilocus ...201627747194
economic game theory to model the attenuation of virulence of an obligate intracellular bacterium.diseases induced by obligate intracellular pathogens have a large burden on global human and animal health. understanding the factors involved in the virulence and fitness of these pathogens contributes to the development of control strategies against these diseases. based on biological observations, a theoretical model using game theory is proposed to explain how obligate intracellular bacteria interact with their host. the equilibrium in such a game shows that the virulence and fitness of the ...201627610355
cellular immune responses induced <i>in vitro</i> by <i>ehrlichia ruminantium</i> secreted proteins and identification of vaccine candidate peptides.secreted proteins are reported to induce cell-mediated immunity characterised by the production of interferon-gamma (ifn)-γ. in this study three open reading frames (orfs) (erum8060, erum7760, erum5000) encoding secreted proteins were selected from the ehrlichia ruminantium (welgevonden) genome sequence using bioinformatics tools to determine whether they induce a cellular immune response in vitro with mononuclear cells from needle and tick infected animals. the whole recombinant protein of the ...201627608502
molecular diagnosis and genetic diversity of tick-borne anaplasmataceae agents infecting the african buffalo syncerus caffer from marromeu reserve in mozambique.tick-borne diseases (tbds) are very important in relation to domestic ruminants, but their occurrence among wild ruminants, mainly in the african buffalo syncerus caffer, remains little known.201627531003
anaplasma, ehrlichia and rickettsial pathogens in ixodid ticks infesting cattle and sheep in western oromia, ethiopia.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 dete ...201627411938
ehrlichia ruminantium infects rhipicephalus microplus in west africa.the invasion of west africa by rhipicephalus microplus during the past decade has changed the ecological situation of the agent of heartwater ehrlichia ruminantium in this area. before, its local vector, amblyomma variegatum, was the most abundant tick species found on livestock. today, the abundance of the r. microplus is one magnitude higher than that of a. variegatum in many west-african localities. we investigated the potential of this new ecological situation to impact the circulation of e. ...201627334439
draft genome sequences of three strains of ehrlichia ruminantium, a tick-borne pathogen of ruminants, isolated from zimbabwe, the gambia, and ghana.the rickettsial bacterium ehrlichia ruminantium is the causative pathogen of heartwater in ruminants. here, we report the draft genome sequences of three strains of e. ruminantium, namely, the crystal springs strain from zimbabwe, the kerr seringe strain from the gambia, and the sankat 430 strain from ghana.201627313287
tick-borne pathogens of zoonotic and veterinary importance in nigerian cattle.ticks and tick-borne diseases undermine cattle fitness and productivity in the whole of sub-saharan africa, including nigeria. in this west african country, cattle are challenged by numerous tick species, especially during the wet season. consequently, several tbds are known to be endemic in nigerian cattle, including anaplasmosis, babesiosis, cowdriosis and theilerioris (by theileria mutans and theileria velifera). to date, all investigations on cattle tbds in nigeria have been based on cytolog ...201627090756
molecular epidemiological surveillance to assess emergence and re-emergence of tick-borne infections in tick samples from china evaluated by nested pcrs.an investigation was performed to detect eight pathogens in ticks collected from grass tips or animals in the southern, central and northeast regions of china. dna samples extracted from ticks were collected from ten different locations in eight provinces of china and subjected to screening for tick-borne pathogens, including borrelia burgdorferi sensu lato, ehrlichia spp., rickettsia spp., babesia/theileria spp., ehrlichia ruminantium, coxiella burnetii, and francisella tularensis, using nested ...201626943995
molecular screening for anaplasmataceae in ticks and tsetse flies from ethiopia.hard ticks and tsetse flies are regarded as the most important vectors of disease agents in sub-saharan africa. with the aim of screening these blood-sucking arthropods for vector-borne pathogens belonging to the family anaplasmataceae in south-western ethiopia, four species of tsetse flies (collected by traps) and seven species of ixodid ticks (removed from cattle) were molecularly analysed. dna was extracted from 296 individual ticks and from 162 individuals or pools of tsetse flies. besides k ...201626919143
growth of ehrlichia canis, the causative agent of canine monocytic ehrlichiosis, in vector and non-vector ixodid tick cell lines.canine monocytic ehrlichiosis is caused by ehrlichia canis, a small gram-negative coccoid bacterium that infects circulating monocytes. the disease is transmitted by the brown dog tick rhipicephalus sanguineus s.l. and is acknowledged as an important infectious disease of dogs and other members of the family canidae worldwide. e. canis is routinely cultured in vitro in the canine monocyte-macrophage cell line dh82 and in non-vector ixodes scapularis tick cell lines, but not in cells derived from ...201626837859
seroprevalence of ehrlichia ruminantium antibodies and its associated risk factors in indigenous goats of south africa.the present study investigated the seroprevalence of antibodies to ehrlichia ruminantium and the associated risk factors in goats from five different farming provinces of south africa. sera collected from 686 goats of the commercial meat type (n=179), mohair type (n=9), non-descript indigenous goats from eastern cape (n=56), kwazulu-natal (n=209), limpopo (n=111), north west (n=61) and northern cape (n=11) provinces and a feral tankwa goat (n=50) were tested for the presence of immunoglobulin g ...201626829905
multiple pathogens including potential new species in tick vectors in côte d'ivoire.our study aimed to assess the presence of different pathogens in ticks collected in two regions in côte d'ivoire.201626771308
comparative proteomic profiling of ehrlichia ruminantium pathogenic strain and its high-passaged attenuated strain reveals virulence and attenuation-associated proteins.the obligate intracellular bacterium ehrlichia ruminantium (er) causes heartwater, a fatal tick-borne disease in livestock. in the field, er strains present different levels of virulence, limiting vaccine efficacy, for which the molecular basis remains unknown. moreover, there are no genetic tools currently available for er manipulation, thus limiting the knowledge of the genes/proteins that are essential for er pathogenesis and biology. as such, to identify proteins and/or mechanisms involved i ...201526691135
heartwater--ehrlichia ruminantium infection.summary heartwater is a notifiable disease that is listed by the world organisation for animal health. it is caused by ehrlichia ruminantium, an obligately intracellular gram-negative bacterium in the order rickettsiales and the family anaplasmataceae. the disease is borne byticks in the genus amblyomma and causes heartwater, or cowdriosis, in wild and domestic ruminants, primarily in africa, but also in parts of the caribbean. the disease was recognised in south africa in the 19th century and d ...201526601456
geographic range of vector-borne infections and their vectors: the role of african wildlife.the role of african wildlife in the occurrence of vector-borne infections in domestic animals has gained renewed interest as emerging and re-emerging infections occur worldwide at an increasing rate. in africa, biodiversity conservation and the expansion of livestock production have increased the risk of transmitting vector-borne infections between wildlife and livestock. the indigenous african pathogens with transboundary potential, such as rift valley fever virus, african horse sickness virus, ...201526470454
survey of ticks collected from tennessee cattle and their pastures for anaplasma and ehrlichia species.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 eac ...201626348980
molecular detection of zoonotic tick-borne pathogens from ticks collected from ruminants in four south african provinces.ticks carry and transmit a remarkable array of pathogens including bacteria, protozoa and viruses, which may be of veterinary and/or of medical significance. with little to no information regarding the presence of tick-borne zoonotic pathogens or their known vectors in southern africa, the aim of our study was to screen for anaplasma phagocytophilum, borrelia burgdorferi, coxiella burnetii, rickettsia species and ehrlichia ruminantium in ticks collected and identified from ruminants in the easte ...201526227797
multilocus sequence analysis of clinical "candidatus neoehrlichia mikurensis" strains from europe."candidatus neoehrlichia mikurensis" is the tick-borne agent of neoehrlichiosis, an infectious disease that primarily affects immunocompromised patients. so far, the genetic variability of "ca. neoehrlichia" has been studied only by comparing 16s rrna genes and groel operon sequences. we describe the development and use of a multilocus sequence analysis (mlsa) protocol to characterize the genetic diversity of clinical "ca. neoehrlichia" strains in europe and their relatedness to other species wi ...201526157152
molecular-genetic and ultrastructural characteristics of 'candidatus ehrlichia khabarensis', a new member of the ehrlichia genus.recently, a new ehrlichia genetic variant, ehrlichia sp. khabarovsk, was identified in tissue samples of small mammals captured in the russian far east. to further characterize ehrlichia sp. khabarovsk, tissue homogenate from a naturally infected gray red-backed vole (myodes rufocanus) was passaged three times in newborn laboratory mice. using nested pcr ehrlichia sp. khabarovsk dna was detected in tissue samples from infected mice at 1-4 weeks post inoculation. electron microscopic examination ...201526096852
the epidemiology of tick-borne haemoparasites as determined by the reverse line blot hybridization assay in an intensively studied cohort of calves in western kenya.the development of sensitive surveillance technologies using pcr-based detection of microbial dna, such as the reverse line blot assay, can facilitate the gathering of epidemiological information on tick-borne diseases, which continue to hamper the productivity of livestock in many parts of africa and elsewhere. we have employed a reverse line blot assay to detect the prevalence of tick-borne parasites in an intensively studied cohort of indigenous calves in western kenya. the calves were recrui ...201525858115
the role of cd8 t lymphocytes in rickettsial infections.arthropod-borne obligately intracellular bacteria pose a difficult challenge to the immune system. the genera rickettsia, orientia, ehrlichia, and anaplasma evolved mechanisms of immune evasion, and each interacts differently with the immune system. the roles of cd8 t cells include protective immunity and immunopathology. in rickettsia infections, cd8 t cells are protective mediated in part by cytotoxicity toward infected cells. in contrast, tnf-α overproduction by cd8 t cells is pathogenic in l ...201525823954
development of a quantitative pcr assay for differentiating the agent of heartwater disease, ehrlichia ruminantium, from the panola mountain ehrlichia.panola mountain ehrlichia (pme) is an emerging ehrlichia sp. reported in ten us states. based on the sequence homology of all known genes, pme is closely related to ehrlichia ruminantium (er), the causative agent of heartwater. heartwater is an economically important tick-borne disease of cattle, sheep and goats responsible for stock losses in sub-saharan africa. unfortunately, er was imported to the caribbean islands in the 19th century, and the presence of this foreign animal disease in the ca ...201625807955
development of a new pcr-based assay to detect anaplasmataceae and the first report of anaplasma phagocytophilum and anaplasma platys in cattle from algeria.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 ba ...201525748051
proteomic profiling of the outer membrane fraction of the obligate intracellular bacterial pathogen ehrlichia ruminantium.the outer membrane proteins (omps) of gram-negative bacteria play a crucial role in virulence and pathogenesis. identification of these proteins represents an important goal for bacterial proteomics, because it aids in vaccine development. here, we have developed such an approach for ehrlichia ruminantium, the obligate intracellular bacterium that causes heartwater. a preliminary whole proteome analysis of elementary bodies, the extracellular infectious form of the bacterium, had been performed ...201525710494
tick-borne pathogens of potential zoonotic importance in the southern african region.the aim of this communication is to provide preliminary information on the tick-borne pathogens of potential zoonotic importance present in southern africa, mainly focusing on their geographical distribution and host range, and to identify research gaps. the following tick-borne zoonoses have been reported to occur in southern africa based mainly on case reports: crimean-congo haemorrhagic fever caused by crimean-congo haemorrhagic fever virus; ehrlichiosis caused by ehrlichia ruminantium, ehrli ...201425685942
tick-borne haemoparasites in african buffalo (syncerus caffer) from two wildlife areas in northern botswana.the african buffalo (syncerus caffer) is a host for many pathogens known to cause economically important diseases and is often considered an important reservoir for livestock diseases. theileriosis, heartwater, babesiosis and anaplasmosis are considered the most important tick-borne diseases of livestock in sub-saharan africa, resulting in extensive economic losses to livestock farmers in endemic areas. information on the distribution of tick-borne diseases and ticks is scarce in northern botswa ...201525589064
a user-friendly and scalable process to prepare a ready-to-use inactivated vaccine: the example of heartwater in ruminants under tropical conditions.the use of cheap and thermoresistant vaccines in poor tropical countries for the control of animal diseases is a key issue. our work aimed at designing and validating a process for the large-scale production of a ready-to-use inactivated vaccine for ruminants. our model was heartwater caused by the obligate intracellular bacterium ehrlichia ruminantium (er). the conventional inactivated vaccine against heartwater (based on whole bacteria inactivated with sodium azide) is prepared immediately bef ...201525514207
improved molecular detection of ehrlichia and anaplasma species applied to amblyomma ticks collected from cattle and sheep in ethiopia.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 an ...201525438799
coinfection of tick cell lines has variable effects on replication of intracellular bacterial and viral pathogens.ticks transmit various human and animal microbial pathogens and may harbour more than one pathogen simultaneously. both viruses and bacteria can trigger, and may subsequently suppress, vertebrate host and arthropod vector anti-microbial responses. microbial coinfection of ticks could lead to an advantage or disadvantage for one or more of the microorganisms. in this preliminary study, cell lines derived from the ticks ixodes scapularis and ixodes ricinus were infected sequentially with 2 arthrop ...201424685441
an assessment of zoonotic and production limiting pathogens in rusa deer (cervus timorensis rusa) from mauritius.a population of approximately 70,000 rusa deer (cervus timorensis russa) represents the most important mammal species reared for food on the island of mauritius, being the main source of red meat for the local population. however, very limited information is available on the circulation of pathogens affecting the productivity and health of this species. to produce baseline data on the circulation of infectious pathogens in rusa deer under production, a serological survey and/or direct pathogen d ...201424382104
novel ehrlichia and hepatozoon agents infecting the crab-eating fox (cerdocyon thous) in southeastern brazil.this study evaluated infection by vector-borne agents in 58 crab-eating fox (cerdocyon thous l.) that were road-killed in an atlantic rainforest reserve in the state of espírito santo, southeastern brazil. spleen, lung, or blood samples collected from the foxes were tested in the laboratory by a battery of polymerase chain reaction (pcr) assays targeting bacteria of the genera rickettsia, borrelia, coxiella, anaplasma, and ehrlichia; and protozoa of the genera babesia, hepatozoon, and leishmania ...201323802461
opportunities in diagnostic and vaccine approaches to mitigate potential heartwater spreading and impact on the american mainland.heartwater, caused by the rickettsiales ehrlichia ruminantium (er), is a tropical tick-borne disease of wild and domestic ruminants, transmitted by amblyomma ticks. it causes significant economic losses due to high mortality and the high cost of antibiotic treatment of affected animals, limiting herd productivity. it is present in sub-saharan africa, islands in the indian ocean and two caribbean islands (guadeloupe and antigua) from where it threatens the american mainland due to risk of the spr ...201323689897
survey of anaplasmataceae bacteria in sheep from senegal.the authors studied the role of bacteria belonging to anaplasmataceae family as the causes of acute illnesses of sheep in west africa.201323553260
identification and characterization of sialidase-like activity in the developmental stages of amblyomma variegatum.amblyomma variegatum f. are obligate hematophagous ectoparasites of livestock that serve as the vectors of ehrlichia ruminantium (formerly known as cowdria ruminantium), the causative agent of heartwater disease. in the light of the fact that they are blood-feeding, their salivary glands play prominent role in their acquisition of nutrients from the bloodmeal. sialic acids are a major component of glycoprotein in mammalian blood fluid and cells. sialome of hard ticks is still sparse. here, for t ...201323427656
theileria, babesia, and anaplasma detected by pcr in ruminant herds at bié province, angola.distribution of anaplasma spp., babesia spp., theileria spp., and ehrlichia ruminantium, was for the first time studied in bié province, central angola. we examined 76 blood samples of cattle originated from seven farms, and 13 blood samples of goats from two farms employing molecular genetic tools (pcr). most prevalent was a. ovis-infection in goats (100%) and a. marginale-infection in cattle (38% of examined animals, and six out of seven farms). b. bigemina-infection was detected in only one s ...023193527
prevalence of ehrlichia ruminantium in adult amblyomma variegatum collected from cattle in cameroon.ehrlichia ruminantium, the etiologic agent of the economically important disease heartwater, is an obligate intracellular bacterium transmitted by ticks of the genus amblyomma, particularly a. hebraeum and a. variegatum. although serologic and microscopic evidence of the presence of heartwater have been reported in ruminants in cameroon, knowledge of e. ruminantium infection in the tick vector, a. variegatum, is lacking. in order to determine the infectivity of a. variegatum ticks by e. ruminant ...201322833099
ehrlichia species, probable emerging human pathogens in sub-saharan africa: environmental exacerbation.ehrlichiae are obligate intracellular gram-negative tick-borne bacteria that are responsible for life-threatening emerging human zoonoses and diseases of veterinary importance worldwide, collectively called ehrlichioses. the genus ehrlichia consists of five recognized species, including e. canis, e. chaffeensis, e. ewingii, e. muris, and e. ruminantium. the recent discoveries of ehrlichia species in new areas and of tick species that were previously thought to be uninfected by these agents have ...201122435325
biological response of endothelial cells and its modulation by cytokines: prospects for therapy and bioprocesses.endothelial cells are involved in important pathological situations. they could be the target for infectious processes as for example in cowdriosis, an important disease in cattle due to the rickettsia cowdria ruminantium prevalent in the south of the sahara. they are also connected to angiogenic processes related to tumor invasion.our results indicate that aids related kaposi sarcoma cells may be of endothelial origin. we conclude from our data the mobility of those cells, related to the expres ...199722358890
ticks and tick-borne pathogens in livestock from nomadic herds in the somali region, ethiopia.between may 2006 and january 2007, blood samples and ticks were randomly collected from 220 nomadic animals from filtu and dollo odo districts, libaan zone, in the somali region of ethiopia. overall, 81.5% cattle, 98.2% camels, 53.4% goats and 61.1% sheep were infested by ixodid ticks. collected ticks (n = 1,036) were identified as rhipicephalus pulchellus (40.1%), r. pravus (25.8%), amblyomma gemma (9.4%), hyalomma rufipes (13.3%), h. truncatum (2.8%), h. impeltatum (1.2%) and h. dromedarii (0. ...201222349943
identification of ehrlichia ruminantium proteins that activate cellular immune responses using a reverse vaccinology strategy.ehrlichia ruminantium is an obligate intracellular bacterial pathogen which causes heartwater, a serious tick-borne disease of ruminants throughout sub-saharan africa. the development of promising recombinant vaccines has been reported previously, but none has been as effective as immunisation with live organisms. in this study we have used reverse vaccinology to identify proteins that elicit an in vitro cellular immune response similar to that induced by intact e. ruminantium. the experimental ...201122261504
Proteomic analyses of Ehrlichia ruminantium highlight differential expression of MAP1-family proteins.The Rickettsiales Ehrlichia ruminantium (ER) is the causative agent of heartwater, a fatal tick-borne disease of livestock in sub-Saharan Africa and in the Caribbean, posing strong economical constraints to livestock production. In an attempt to identify the most prominent proteins expressed by this bacterium, especially those encoded by the major antigenic protein 1 (map1) multigene family, a proteome map of ER cultivated in endothelial cells was constructed by using two dimensional gel electro ...201122204792
survey of ixodid ticks and two tick-borne pathogens in african buffaloes, syncerus caffer, from the caprivi strip, namibia.a capture operation to ascertain health status in free-ranging buffaloes from six different areas in the caprivi strip in the northeast corner of namibia was conducted in october 2009. basic information on the ticks and tick-borne pathogens normally found in wildlife from this area are scarce. the objective of this study was to assess the host status of african buffaloes, syncerus caffer, for ixodid ticks and two selected tick-borne pathogens in the caprivi strip, a key area bordering angola, za ...201122204058
A new typing technique for the Rickettsiales Ehrlichia ruminantium: Multiple-locus variable number tandem repeat analysis.Ehrlichia ruminantium (ER) is a member of the order Rickettsiales transmitted by Amblyomma ticks. This obligatory intracellular bacterium is the causative agent of a fatal disease in ruminants, named heartwater. It represents a constraint on breeding development in sub-Saharan Africa and in the Caribbean. The genetic diversity of the strains of ER, which could be a limiting factor to obtain effective vaccines, needs to be better characterized. For this purpose, we developed a molecular typing te ...201122143037
Global gene expression profiling of Ehrlichia ruminantium at different stages of development.Ehrlichia ruminantium (ER), the causative agent of heartwater on ruminants, is an obligate intracellular bacterium transmitted by ticks of the genus Amblyomma. Previous studies have shown that early stages of development may be critical for Ehrlichia pathogenicity. To gain insights into the biology of intracellular ER, we determined the genome-wide transcriptional profile of ER replicating inside bovine aortic endothelial cells using DNA microarrays. At intermediate and late stages of infection ...201122098128
Development of multiple-locus variable-number tandem-repeat analysis for rapid genotyping of Ehrlichia ruminantium and its application to infected Amblyomma variegatum collected in heartwater endemic areas in Uganda.SUMMARYThe rickettsial bacterium Ehrlichia ruminantium is the causative agent of heartwater, a serious tick-borne disease in ruminants. The genetic diversity of organisms in the field will have implications for cross-protective capacities of any vaccine developed, and for an effective vaccine design strategy proper genotyping and understanding of existing genetic diversity in the field is necessary. We searched for variable-number tandem-repeat (VNTR) loci for use in a multi-locus VNTR analysis ...201122008706
preparation and in vitro characterisation of ehrlichia ruminantium plasmid dna and proteins encapsulated into and dna adsorbed onto biodegradable microparticles.four e. ruminantium 1h12 open reading frames and their proteins known to protect sheep against heartwater needle challenge were encapsulated into, or adsorbed onto poly(d,l-lactide-co-glycolide) microparticles. microspheres with smooth surface and smaller than 5 ++m diameters were produced, with high adsorption and encapsulation efficiencies. gel electrophoresis showed that neither encapsulation nor adsorption affected the stability of the dna or proteins. cationic microparticles released ôê+40% ...201021771527
multi-locus sequence typing of ehrlichia ruminantium strains from geographically diverse origins and collected in amblyomma variegatum from uganda.abstract: background: the rickettsial bacterium ehrlichia ruminantium is the causative agent of heartwater in ruminants. a better understanding of the population genetics of its different strains is, however, needed for the development of novel diagnostic tools, therapeutics and prevention strategies. specifically, the development of effective vaccination policies relies on the proper genotyping and characterisation of field isolates. although multi-locus sequence typing (mlst) has been recently ...201121762509
an alternative approach to multiple genome comparison.genome comparison is now a crucial step for genome annotation and identification of regulatory motifs. genome comparison aims for instance at finding genomic regions either specific to or in one-to-one correspondance between individuals/strains/species. it serves e.g. to pre-annotate a new genome by automatically transfering annotations from a known one. however, efficiency, flexibility and objectives of current methods do not suit the whole spectrum of applications, genome sizes and organizatio ...201121646341
tick-borne bacteria in free-living jaguars (panthera onca) in pantanal, brazil.abstract tick-borne bacteria were investigated in 10 free-living jaguars and their ticks in the pantanal biome, brazil. jaguar sera were tested by indirect fluorescent antibody assays using rickettsia rickettsii, rickettsia parkeri, rickettsia amblyommii, rickettsia rhipicephali, rickettsia felis, rickettsia bellii, ehrlichia canis, and coxiella burnetii as crude antigens. all 10 jaguar sera reacted (titer ≥64) to at least one rickettsia species; 4 and 3 sera reacted with e. canis and c. burneti ...201121612532
ehrlichia ruminantium in amblyomma variegatum and domestic ruminants in the caribbean.the highly sensitive nested pcs20 polymerase chain reaction assay for ehrlichia ruminantium was negative on 506 amblyomma variegatum from caribbean islands where clinical heartwater has not been reported, mainly the united states virgin islands (18), dominica (170), montserrat (5), nevis (34), st. kitts (262), and st. lucia (17). positive results were obtained with positive controls (crystal springs strain) and a. variegatum from countries in africa where infections are endemic, mainly tanzania ...201121485394
veterinary extension on sampling techniques related to heartwater research.heartwater, a tick-borne disease caused by ehrlichia ruminantium, is considered to be a significant cause of mortality amongst domestic and wild ruminants in south africa. the main vector is amblyomma hebraeum and although previous epidemiological studies have outlined endemic areas based on mortalities, these have been limited by diagnostic methods which relied mainly on positive brain smears. the indirect fluorescent antibody test (ifa) has a low specificity for heartwater organisms as it cros ...201021247042
treatment and control of chlamydial and rickettsial infections in sheep and goats.small ruminants are susceptible to several chlamydial and rickettsial infections. some of them, such as ehrlichia ruminantium, have a great impact on the sheep and goat industry while others, such as coxiella burnetii, are important zoonotic agents. this review focuses on measures of treatment and control for the following organisms: chlamydophila abortus (formerly chlamydia psittaci immunotype 1), coxiella burnetii, anaplasma ovis, anaplasma phagocytophilum, and ehrlichia ruminantium.201021215905
epidemiological analysis of tick-borne diseases in zambia.tick-borne diseases are a constraint to livestock production in many developing countries as they cause high morbidity and mortality, which results in decreased production of meat, milk and other livestock by-products. the most important tick-borne diseases of livestock in sub-saharan africa are east coast fever (caused by theileria parva), babesiosis (caused by babesia bigemina and b. bovis), anaplasmosis (caused by anaplasma marginale) and heartwater (caused by ehrlichia ruminantium). despite ...201021106294
thoughts on the threat of ehrlichia ruminantium infection. 201021090154
development of loop-mediated isothermal amplification (lamp) assays for rapid detection of ehrlichia ruminantium.the rickettsial bacterium ehrlichia ruminantium is the causative agent of heartwater, a potential zoonotic disease of ruminants transmitted by ticks of the genus amblyomma. the disease is distributed in nearly all of sub-saharan africa and some islands of the caribbean, from where it threatens the american mainland. this report describes the development of two different loop-mediated isothermal amplification (lamp) assays for sensitive and specific detection of e. ruminantium.201021087521
haemoparasite prevalence and theileria parva strain diversity in cape buffalo (syncerus caffer) in uganda.cape buffalo (syncerus caffer) are considered to be an important reservoir for various tick-borne haemoparasites of veterinary importance. in this study we have compared the haemoparasite carrier prevalence in buffalo from four geographically isolated national parks in uganda [lake mburo national park (lmnp), queen elizabeth national park (qenp), murchison falls national park (mfnp) and kidepo valley national park (kvnp)]. differences were seen in haemoparasite prevalence in buffalo from the fou ...201021074945
the gulf coast tick: a review of the life history, ecology, distribution, and emergence as an arthropod of medical and veterinary importance.the gulf coast tick, amblyomma maculatum koch (acari: ixodidae), is a unique univoltine ectoparasite of seven vertebrate host classes in the western hemisphere that is increasingly recognized as a pest of livestock and wildlife, a vector of pathogens to humans and canines, and a putative vector of ehrlichia ruminantium, the causal agent of heartwater, a fatal foreign animal disease of ruminants resident in the caribbean. this review assembles current and historical literature encompassing the bi ...201020939363
recognition of the threat of ehrlichia ruminantium infection in domestic and wild ruminants in the continental united states. 201020807129
a novel ehrlichia genotype detected in naturally infected cattle in north america.during a research investigation to determine if cattle from british columbia (bc), canada were infected with anaplasma marginale or other related rickettsial blood parasites, a novel ehrlichia genotype was revealed. blood from seven bc source cattle was bioassayed by intravenous inoculation into naïve splenectomised calves. additional splenectomised calves were used as uninoculated negative control or a. marginale-inoculated positive control. newly designed sets of primers specific for the msp5 ...201020663613
in vitro and in vivo evaluation of five low molecular weight proteins of ehrlichia ruminantium as potential vaccine components.low molecular weight (lmw) proteins of e. ruminantium can induce proliferation of immune peripheral blood mononuclear cells (pbmcs) and the production of interferon-gamma (ifn-gamma) by cd4+-enriched t-cells. in this study, a reverse vaccinology approach was applied to identify additional vaccine candidates focusing on genes that encode lmw proteins smaller than 20 kda. five open reading frames (orfs) were selected from the e. ruminantium genome and their corresponding recombinant (r) proteins w ...201020566221
clonal origin of emerging populations of ehrlichia ruminantium in burkina faso.cowdriosis or heartwater is a major tick-borne disease on ruminants in africa and the caribbean. the causative agent is ehrlichia ruminantium, an intracellular bacterium. development of vaccines against heartwater has been hampered the limited efficiency of vaccine in the field, thought to be a consequence of the high genetic diversity of strains circulating in a same area. a sampling scheme was set to collect ticks over 2 years in a delimited area and well identified flock. prevalence was low a ...201020553966
efficiency of inactivated vaccines against heartwater in burkina faso: impact of ehrlichia ruminantium genetic diversity.in order to identify the appropriate strains to use in vaccination trials against heartwater in burkina faso, the protective effect of gardel and welgevonden strains was assessed against local strains on sheep vaccinated by infection-and-treatment method: gardel protected significantly against burkina faso strains tested (survival rate 59% for immunised sheep vs 13% for control sheep) while welgevonden did not (survival rate 45% for immunised sheep vs 25% for control sheep). the efficacy of the ...201020470791
heartwater: an abridged historical account.heartwater was first recognised and recorded in south africa as early as 1838. since then the disease has been described from almost all the countries in africa south of the sahara, from certain islands around africa and from a number of islands in the caribbean. most of the research on the disease, at least until fairly recently, was conducted in south africa. progress in research on the disease has been slow but a few important findings are highlighted in this paper.200920458858
use of geographic information systems to identify areas at risk of introducing amblyomma variegatum and a. hebraeum to italy.tick-borne diseases are one of the most important obstacles to the development of livestock breeding in africa. among these diseases, heartwater is of significant importance, second in rank to east coast fever. heartwater affects wild and domestic ruminants and is caused by a micro-organism belonging to the order rickettsiales, previously called cowdria ruminantium and recently renamed ehrlichia ruminantium. the disease is transmitted solely by ticks. although the aetiology and the clinical sign ...200720422545
studies of a polymorphic ehrlichia ruminantium gene for use as a component of a recombinant vaccine against heartwater.a previously identified polymorphic ehrlichia ruminantium gene, erum2510, was investigated to determine its ability to induce protective immunity in ruminants following two different dna immunisation strategies; dna-only and a dna prime/recombinant protein (rprotein) boost immunisation. the dna-only vaccine was also compared to a cocktail of three polymorphic e. ruminantium (welgevonden) open reading frames (orfs) adjacent to erum2510 in the genome. weak protection was observed in animals immuni ...201020338214
science and popular participation in the investigation of heartwater in south africa, c. 1870-1950.during the late nineteenth century, settler farmers in southern africa identified heartwater as a damaging disease of small stock and cattle. they advanced various explanations of the disease, including the theory that it was caused by the bite of ticks. around 1900, the american entomologist c.p. lousbury demonstrated that heartwater was transmitted by the bont tick. he also worked out the life cycle and life habits of the tick. subsequently, farmers developed methods of controlling ticks by di ...200820055238
innovative approach for transcriptomic analysis of obligate intracellular pathogen: selective capture of transcribed sequences of ehrlichia ruminantium.whole genome transcriptomic analysis is a powerful approach to elucidate the molecular mechanisms controlling the pathogenesis of obligate intracellular bacteria. however, the major hurdle resides in the low quantity of prokaryotic mrnas extracted from host cells. our model ehrlichia ruminantium (er), the causative agent of heartwater, is transmitted by tick amblyomma variegatum. this bacterium affects wild and domestic ruminants and is present in sub-saharan africa and the caribbean islands. be ...200920034374
trends in the control of heartwater.heartwater is an economically serious tick-borne disease of ruminants caused by the intracellular bacterium ehrlichia ruminantium. the disease has traditionally been controlled by four different approaches: controlling the tick vector by dipping, establishing endemic stability, performing immunisation by infection and treatment, and preventing the disease by regular administration of prophylactic antibiotics. the first three of these methods are subject to failure for various epidemiological rea ...200919967932
persistence mechanisms in tick-borne diseases.the use of new, highly sensitive diagnostic methods has revealed persistent infections to be a common feature of different tick-borne diseases, such as babesiosis, anaplasmosis and heartwater. antigenic variation can contribute to disease persistence through the continual elaboration of new surface structures, and we know in several instances how this is achieved. known or suspected mechanisms of persistence in babesial parasites include cytoadhesion and rapid variation of the adhesive ligand in ...200919967928
infection of the endothelium by members of the order rickettsiales.the vascular endothelium is the main target of a limited number of infectious agents, rickettsia, ehrlichia ruminantium, and orientia tsutsugamushi are among them. these arthropod-transmitted obligately-intracellular bacteria cause serious systemic diseases that are not infrequently lethal. in this review, we discuss the bacterial biology, vector biology, and clinical aspects of these conditions with particular emphasis on the interactions of these bacteria with the vascular endothelium and how ...200919967137
studies on the etiology of heartwater : iii. the multiplication of rickettsia ruminantium within the endothelial cells of infected animals and their discharge into the circulation.the rickettsiae of heartwater are more definitely restricted to the vascular endothelial cells of infected animals than are those of typhus or rocky mountain spotted fever. they likewise form more pronounced spherical colonies within the cells. their presence does not injure the endothelial cells to a degree at all comparable with that caused by the other pathogenic rickettsiae. the rupture of endothelial cells and discharge of rickettsiae, which are apparently viable, into the circulation const ...192619869225
studies on the etiology of heartwater : ii. rickettsia ruminantium (n. sp.) in the tissues of ticks transmitting the disease.the evidence offered in the first of these studies indicative of a causative relationship between rickettsia ruminantium and heartwater is supplemented by the following observations concerning the ticks which carry the disease. when larvae, which had taken no food since hatching, were allowed to feed upon cases of heartwater, they acquired rickettsioe which appeared to be identical with those in the tissues of animals suffering from heartwater, whereas control larvae hatched from eggs deposited ...192519869050
studies on the etiology of heartwater : i. observation of a rickettsia, rickettsia ruminantium (n. sp.), in the tissues of infected animals.a gram-negative, intracellular, coccus-like microorganism was found in cases of heartwater in the three species which are susceptible to the disease; namely, goats, sheep, and cattle. it was absent in the case of control animals, both normal ones and those dying of some. other diseases. the presence of this microorganism was definitely related to the febrile reaction. it was most easily detected in the renal glomeruli and in the small capillaries of the cerebral cortex but probably occurred thro ...192519869049
natural history of ehrlichia ruminantium.ehrlichia ruminantium is an obligately intracellular proteobacterium which causes a disease known as heartwater or cowdriosis in some wild, and all domestic, ruminants. the organism is transmitted by ticks of the genus amblyomma, and it is of serious economic importance wherever the natural vectors occur, an area which includes all of sub-saharan africa, and several islands in the caribbean. the disease was first recognized in south africa in the 19th century, where its tick-borne nature was det ...201019836892
mining the genetic diversity of ehrlichia ruminantium using map genes family.understanding bacterial genetic diversity is crucial to comprehend pathogenesis. ehrlichia ruminantium (e. ruminantium), a tick-transmitted intracellular bacterial pathogen, causes heartwater disease in ruminants. this model rickettsia, whose genome has been recently sequenced, is restricted to neutrophils and reticulo-endothelial cells of its mammalian host and to the midgut and salivary glands of its vector tick. e. ruminantium harbors a multigene family encoding for 16 outer membrane proteins ...201019819629
seroprevalence of tick-borne diseases in communal cattle reared on sweet and sour rangelands in a semi-arid area of south africa.a survey was conducted to determine the seroprevalence of tick-borne diseases (tbd) in nguni and non-descript cattle on the sweet and sour communal rangelands across seasons. body condition scores, bodyweights, packed cell volume (pcv) and antibodies to b. bovis, b. bigemina, ehrlichiaruminatium and anaplasma marginale were determined seasonally in 144 cattle raised on communal rangelands from august 2007 to april 2008. approximately 45% of the cattle were seropositive for b. bovis, 46% for b. b ...201019733490
mlst scheme of ehrlichia ruminantium: genomic stasis and recombination in strains from burkina-faso.heartwater, caused by the intracellular bacterium ehrlichia ruminantium, is a major tick-borne disease of livestock in africa also introduced in the caribbean. the main problem encountered with the control of this disease is the lack of efficient vaccine in the field. this is thought to be related to the high genetic diversity of strains circulating in a same area. a set of eight circulating strains was isolated from a herd of cows in a small locality in burkina-faso and analyzed along with two ...200919712754
new approaches to detection and identification of rickettsia africae and ehrlichia ruminantium in amblyomma variegatum (acari: ixodidae) ticks from the caribbean.imported from africa in the 1700s and despite frequent modern eradication efforts, amblyomma variegatum (f.) spread through the caribbean by cattle transport, small ruminants, and migrating birds. a. variegatum is a vector for rickettsia africae, the causative agent of african tick bite fever, and ehrlichia ruminantium, the causative agent of heartwater. we examined 95 a. variegatum and six rhipicephalus (boophilus) microplus (canestrini) collected from cattle at an abattoir in antigua. engorged ...200919645301
diversity of ehrlichia ruminantium major antigenic protein 1-2 in field isolates and infected sheep.proteins expressed from the map1 multigene family of ehrlichia ruminantium are strongly recognized by immune t and b cells from infected animals or from animals that were infected and have recovered from heartwater disease (although still remaining infected carriers). analogous multigene clusters also encode the immunodominant outer membrane proteins (omps) in other ehrlichial species. recombinant protein analogs of the expressed genes and dna vaccines based on the multigene clusters have been s ...200919307215
experimental use of the attenuated ehrlichia ruminantium (welgevonden) vaccine in merino sheep and angora goats.the attenuated ehrlichia ruminantium (welgevonden) stock provides protection against a virulent homologous needle challenge in merino sheep and boer goats against heartwater. in this study, cryopreserved stabilates were tested for their suitability as a vaccine in merino sheep. vaccination did not produce disease and upon challenge with the virulent homologous stock all animals were fully protected. the vaccination protected all except one animal out of 5 challenged 12 months after immunization. ...200819178891
attenuated vaccines for animal diseases. 200819178885
toward prevention of cowdriosis.mass production of ehrlichia ruminantium variants from different regions of sub-saharan africa is one of the difficulties that must be overcome in producing a heartwater vaccine. vaccine productivity can be limited by endogenous induction of interferon (ifn), which inhibits the propagation of ehrlichia ruminantium (er) in cell culture. different kinds of endothelial cells, in which er multiply efficiently, could be grown in a scalable way in vuelife teflon bags on cytodex 3 microcarriers where b ...200819120230
amblyomma variegatum ticks and heartwater on three caribbean islands.amblyomma variegatum tick infestation, tick infection by ehrlichia ruminantium (er), and er genetic diversity were studied in the caribbean islands of guadeloupe, marie-galante, and antigua between 2003 and 2005. nested pcr for pcs20 was used to detect er, while er strains were characterized by sequencing or by restriction fragment length polymorphism (rflp) profiles of map-1 pcr products. in 2003 in guadeloupe, the prevalence of tick-infested herds was 35.6%. in marie-galante 79.1% of herds in ...200819120208
serological survey of antibodies to ehrlichia ruminantium in small ruminants in tanzania.sera from 497 sheep and 555 goats collected in a cross sectional study from different geographical locations in north-eastern tanzania were examined for antibodies to ehrlichia ruminantium using map 1-b elisa technique. e. ruminantium antibodies were found in 68.6% (341/497) of sheep and 64.7% (359/555) of goats. overall seroprevalence was 66.5% (700/1052). infection rates were higher in sheep than goats (p < 0.05), in pastoral than in agro-pastoral production systems (p < 0.05) and in female sh ...200919067218
the prevalence of serum antibodies to ehrlichia ruminantium infection in ranch cattle in tanzania: a cross-sectional study.serum samples collected in a cross-sectional survey of grazing cattle on manyara ranch, monduli district, tanzania, were tested by indirect major antigenic protein 1 fragment b (map 1-b) elisa to determine the seroprevalence of ehrlichia ruminantium and to assess ranch-level risk factors for heartwater. heartwater-exposed cattle were widespread on the ranch and overall seroprevalence was 50.3% (95% ci, 44.9-55.6), enough to indicate an endemically unstable situation. multivariate logistic regres ...200818846851
in vitro isolation of ehrlichia ruminantium from ovine blood into ixodes scapularis (ide8) cell cultures.four stocks of ehrlichia ruminantium (welgevonden, ball3, nonile and blaauwkrans), the causative agent of heartwater in domestic ruminants, were isolated into ixodes scapularis (ide8) tick cells using the leukocyte fraction of the blood of infected sheep. organisms of two of the e. ruminantium stocks (welgevonden and blaauwkrans) propagated in ide8 cells were also successfully used to infect bovine endothelial cells. all stocks were successfully propagated in ide8 cells using dulbecco's modified ...200818788205
a heterologous prime/boost immunisation strategy protects against virulent e. ruminantium welgevonden needle challenge but not against tick challenge.heterologous prime/boost immunisation strategies using the ehrlichia ruminantium 1h12 pcmviubs_orfs [pretorius a, collins ne, steyn hc, van strijp f, van kleef m, allsopp ba. protection against heartwater by dna immunisation with four ehrlichia ruminantium open reading frames. vaccine 2007;25(12):2316-24] were investigated in this study. all the animals immunised twice with a recombinant (r) dna cocktail of four 1h12 pcmviubs_orfs followed by a r1h12 protein and those immunised 3x with 1h12 plas ...200818602727
a quantitative real-time pcr assay for ehrlichia ruminantium using pcs20.heartwater is a tick borne disease that affects ruminants and wild animals in africa south of the sahara. it is caused by ehrlichia ruminantium and transmitted by the tick amblyomma hebraeum. the protocols currently used to detect heartwater take several days to complete. here, we describe the development of a pcs20 quantitative real-time pcr taqman probe assay to detect e. ruminantium in livestock blood and ticks from the field. the assay is based on the conserved pcs20 gene region of e. rumina ...200818511221
natural and experimental infection of white-tailed deer (odocoileus virginianus) from the united states with an ehrlichia sp. closely related to ehrlichia ruminantium.an ehrlichia sp. (panola mountain [pm] ehrlichia sp.) closely related to ehrlichia ruminantium was recently detected in a domestic goat experimentally infested with lone star ticks (lsts, amblyomma americanum) collected from georgia, usa. the infected goat exhibited pyrexia and mild clinical pathologic abnormalities consistent with ehrlichiosis. at least two other ehrlichia species (ehrlichia chaffeensis and ehrlichia ewingii) are maintained in nature by a cycle involving lsts as the primary vec ...200818436670
geographic distribution and genetic diversity of the ehrlichia sp. from panola mountain in amblyomma americanum.a novel ehrlichia, closely related to ehrlichia ruminantium, was recently discovered from panola mountain state park, ga, usa. we conducted a study to determine if this agent was recently introduced into the united states.200818433500
amblyomma variegatum in cattle in marie galante, french antilles: prevalence, control measures, and infection by ehrlichia ruminantium.we report marie galante as one of the caribbean islands most heavily infested by the tropical bont tick (tbt) amblyomma variegatum which is associated with two major diseases of ruminants: heartwater and dermatophilosis. in 2005, a survey was undertaken to assess the prevalence of tbt infestation in cattle, the prevalence of ehrlichia ruminantium infection in tbts, and the tick control measures implemented by livestock owners. a random sample of 195 cattle herds out of 1885 recorded on the islan ...200818406061
climate changes and suitability for the ticks amblyomma hebraeum and amblyomma variegatum (ixodidae) in zimbabwe (1974-1999).the spread of amblyomma hebraeum has been reported in zimbabwe. at the same time there was little or no spread in the distribution of amblyomma variegatum. this paper examines the climatic cycles and their trends in the period 1974-1999 with a view to explaining the abiotic causes of this spread, and of forecasting the likely tendency in climate suitability for both tick species. an annual data-set of rainfall and air temperature was used as a source for climate, together with a habitat-modeling ...200818083309
host cell-specific protein expression in vitro in ehrlichia ruminantium.ehrlichia ruminantium, a tick-transmitted pathogen, is the causative agent of heartwater in ruminants. in this study, a proteomic approach was used to identify host cell-specific e. ruminantium proteins encoded by the map1 multigene family, expressed in vitro in bovine endothelial and tick cell cultures. two-dimensional gel electrophoresis combined with mass spectrometry analysis was used to establish the identities of immunodominant proteins. proteins extracted from e. ruminantium-infected endo ...200818006251
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