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PubMed1999

Treatment of East Coast fever: a comparison of parvaquone and buparvaquone.

Two trials were conducted to compare the efficacy of parvaquone and buparvaquone for the treatment of naturally acquired East Coast fever (ECF, Theileria parva infection) which, if untreated, is almost invariably fatal in European breeds of cattle. In the first trial 28 naive cattle were exposed in a paddock infested with ticks carrying a virulent form of the disease. Twelve were treated with each drug when they developed clinical ECF. All 24 cattle were cured. In the second study, 100 cases of ECF occurring naturally on farms in Kenya were treated, 50 with each drug. Parvaquone cured 44 (88%) buparvaquone cured 45 (90%). Intercurrent infections, predominantly anaplasmosis and bacterial pneumonia or scour, were treated specifically. It is concluded that parvaquone and buparvaquone are similarly effective in curing ECF and cure rates are maximised by accurate diagnosis and prompt treatment of both ECF and intercurrent infections.

Administration, Topical↗
PubMed2000

Sequence and expression analysis of a surface antigen gene family of the rickettsia Anaplasma marginale.

The tick-borne rickettsial organism, Anaplasma marginale, causes a disease in cattle of world-wide economic significance. This disease, anaplasmosis, is characterized by severe hemolytic anemia, high levels of rickettsemia and, often, death in animals over 3years of age. Animals that survive acute infection remain carriers, with continuous sub-microscopic cycles of rickettsemia that can persist for the lifetime of the animal. In the search for potential recombinant immunogens, it was discovered that several surface proteins of A. marginale encode polymorphic multigene families. Despite the small size of the genome (approx. 1250kb), these surface antigen gene families comprise greater than 2% of the genome. We present here a mapping, sequencing and expression analysis of five complete or partial genes encoding MSP1b in a Florida strain of A. marginale. Two genes are complete; they encode mRNA that is translated into polypeptide products. Three genes are incomplete and appear to be derived from the complete genes by a series of segmental intragenic recombinations. In two of the incomplete genes, 5' sequence in the incomplete genes is 3' sequence in the complete genes. Recombination within these gene families may generate diversity in surface antigens through combinatorial rearrangements. This could contribute to persistence in the chronic infections caused by A. marginale and related rickettsiae.

Anaplasma↗
PubMed2003

Vector-borne diseases in humans and animals: activities of the Swiss Tropical Institute and risks for Switzerland.

The recent outbreak of anaplasmosis in a Swiss cattle herd triggered a discussion of the risk of vector-borne diseases in animals and humans in relation to climate changes and other factors. This overview presents the Swiss Tropical Institute's (STI) activities on vector-borne diseases (malaria, trypanosomosis, and leishmaniosis in humans and tick-borne diseases in livestock), describes the possible risks for humans and animals in Switzerland, and discusses options for action in the domains of public health, livestock production and companion animals. Switzerland is increasingly confronted with vector-borne diseases in humans and animals, but this is mainly due to an increase in imported cases. The emergence of a disease in one sector (human or veterinary medicine) may predict future trends in the other. A stronger intersectoral collaboration between public health and veterinary institutions at the federal and cantonal level is needed.

Animal Husbandry↗
PubMed2006

[Piroplasms of ruminants in Switzerland and zoonotic significance of Babesia].

Piroplasms are tick-transmitted blood parasites belonging to the genera Babesia and Theileria. In western and southern Switzerland, B. divergens, a small Babesia species, has been known for a long time as a parasite of cattle. Recent investigations have revealed the autochthonous occurrence of this parasite also in central and eastern Switzerland. On the occasion of an outbreak of anaplasmosis in the canton of Grisons, however, B. bigemina, a large Babesia species, and Theileria of the buffeli/sergenti/orientalis species complex were for the first time identified; the epidemiology of these two piroplasms in Switzerland remains unknown until now. The recent identification by genetic analyses of B. divergens in wild ruminants contradicts the hitherto postulated strict host specificity of this Babesia species for cattle. B. divergens as well as the closely related Babesia spp. genotype EU1 have in single cases also been identified in splenectomized humans. The rodent babesia B. microti which causes a human infection that is considered an "emerging tick-borne disease" in the U.S.A., is widespread in rodent populations in Switzerland, but seems to be of minor relevance as zoonotic pathogen here. Reasons for this could be differences in virulence of the parasites or in the transmission by the respective tick-vectors on the two continents.

Animals↗
PubMed2001

Efficient use of a small genome to generate antigenic diversity in tick-borne ehrlichial pathogens.

Ehrlichiae are responsible for important tick-transmitted diseases, including anaplasmosis, the most prevalent tick-borne infection of livestock worldwide, and the emerging human diseases monocytic and granulocytic ehrlichiosis. Antigenic variation of major surface proteins is a key feature of these pathogens that allows persistence in the mammalian host, a requisite for subsequent tick transmission. In Anaplasma marginale pseudogenes for two antigenically variable gene families, msp2 and msp3, appear in concert. These pseudogenes can be recombined into the functional expression site to generate new antigenic variants. Coordinated control of the recombination of these genes would allow these two gene families to act synergistically to evade the host immune response.

Amino Acid Sequence↗
PubMed1988

Detection of Anaplasma-marginale-infected tick vectors by using a cloned DNA probe.

Anaplasmosis is the most widely distributed of several important tick-borne diseases that constrain cattle production throughout much of the world. Evaluation of the effectiveness of disease control strategies that integrate vaccination with tick control requires the ability to monitor tick and cattle infection rates. To detect Anaplasma marginale in ticks and bovine erythrocytes, a 2-kilobase DNA fragment from a cloned A. marginale gene coding for a surface protein having a Mr of 105,000 was prepared and evaluated as a probe. The probe was species specific and detected A. marginale DNA derived from infected bovine erythrocytes and adult Dermacentor ticks infected either as nymphs or adults. Tick infection was confirmed by microscopy and test feeding on a susceptible calf. The sensitivity of the probe is suitable for detecting infected ticks in experimental and field epizootiology studies.

Anaplasma↗
PubMed1992

Application of a monoclonal antibody-based antigen detection enzyme-linked immunosorbent assay (antigen ELISA) for field diagnosis of bovine trypanosomiasis at Nguruman, Kenya.

A monoclonal antibody-based, enzyme immunoassay (antigen ELISA) for the detection of species-specific invariant antigens of Trypanosoma congolense, T. vivax or T. brucei in the serum of infected animals was evaluated as a means of diagnosis using bovine field sera from a trypanosomiasis endemic area, Nguruman, Kenya. Circulating trypanosome antigens were detected in 126 (96.2%) of 131 serum samples from animals with parasitologically confirmed diagnosis: 74.8% were positive for antigens of two or three trypanosome species, while 21.4% tested positive for one trypanosome species. When 70 sera from animals (at Nguruman), which had tested negative for trypanosomes by the buffy coat technique, were tested, 35 (50.0%) of them were shown to be antigen-ELISA positive: 24 (34.3%) showing infection with a single species and 11 (15.7%) with mixed infections. The predominant trypanosome species diagnosed in the two herds by antigen ELISA was T. vivax, which was detected in 133 (82.6%) of the 161 sera that tested positive for antigens, followed by T. congolense in 122 (75.8%) sera, with 109 (67.7%) showing evidence of mixed infections with two or three trypanosome species. In single infections, T. vivax exceeded T. congolense by a ratio of 2:1, with T. brucei accounting for less than 1.0%. Evidence for the specificity of the test was provided by analysis of field sera from 100 cattle, from a trypanosomiasis-free area, infected with other haemoparasites (anaplasmosis, babesiosis and theileriosis), which all tested negative in the assay.

Animals↗
PubMed1965

The nature of bands in parasitized bovine erythrocytes.

Anaplasma marginale is the etiological agent of a hemolytic disease of cattle, known as anaplasmosis. The organism appears as a marginal inclusion in parasitized erythrocytes, but certain isolates also have bands associated with the inclusion. Inclusions and associated bands in parasitized erythrocytes in the liver and peripheral circulation were studied by light microscope cytochemistry and electron microscopy. Bands were comet- and dumbbell-shaped by light microscopy and were stained by techniques used to demonstrate protein and fibrin. The same forms, as well as other shapes, were seen in infected erythrocytes which were sectioned and examined by electron microscopy. Bands had longitudinal and transverse periodicity. They did not appear to have a crystalline structure. Their appearance was collated with that of bovine fibrin. Bands were well differentiated in erythrocytes that were entensively hemolyzed by natural or artificial means, but poorly differentiated in mildly hemolyzed erythrocytes. Hemolysis methods appeared to influence the morphology of bands and their demonstration.

Anaplasma↗
PubMed2006

An Ixodes scapularis protein required for survival of Anaplasma phagocytophilum in tick salivary glands.

Anaplasma phagocytophilum is the agent of human anaplasmosis, the second most common tick-borne illness in the United States. This pathogen, which is closely related to obligate intracellular organisms in the genera Rickettsia, Ehrlichia, and Anaplasma, persists in ticks and mammalian hosts; however, the mechanisms for survival in the arthropod are not known. We now show that A. phagocytophilum induces expression of the Ixodes scapularis salp16 gene in the arthropod salivary glands during vector engorgement. RNA interference-mediated silencing of salp16 gene expression interfered with the survival of A. phagocytophilum that entered ticks fed on A. phagocytophilum-infected mice. A. phagocytophilum migrated normally from A. phagocytophilum-infected mice to the gut of engorging salp16-deficient ticks, but up to 90% of the bacteria that entered the ticks were not able to successfully infect I. scapularis salivary glands. These data demonstrate the specific requirement of a pathogen for a tick salivary protein to persist within the arthropod and provide a paradigm for understanding how Rickettsia-like pathogens are maintained within vectors.

Anaplasma phagocytophilum↗
PubMed2006

Evaluation of white-tailed deer (Odocoileus virginianus) as natural sentinels for Anaplasma phagocytophilum.

Anaplasma phagocytophilum, the causative agent of human granulocytotropic anaplasmosis, can infect white-tailed deer (WTD; Odocoileus virginianus), and this species is a crucial host for adult Ixodes scapularis, the primary vector of A. phagocytophilum. The goal of this study was to determine the geographic distribution of A. phagocytophilum among WTD across a 19 state region and to evaluate the utility of WTD as natural sentinels. Serologic testing using the indirect fluorescent antibody (IFA) assay was conducted on WTD serum samples and molecular and xenodiagnostic tests were performed to confirm serologic results. The surveillance system was assessed through examination of vital attributes including WTD age and gender associations with serologic status, sample size adequacy for accurate infection status classification, and presence of the vector, I. scapularis. Six hundred thirty-three of 2,666 (24%) WTD in 17 states tested positive for antibodies (>or=128) when tested by IFA assay. Testing for p44 and/or 16S rRNA gene targets identified 73 (16%) PCR positive WTD among 458 animals tested, all of which originated from seropositive populations. Attempts to culture A. phagocytophilum from WTD were unsuccessful; however, xenodiagnostic mice inoculated with blood from 3 WTD became infected. Seroprevalence did not differ by deer age or gender; however, WTD<or=0.75 years old had a higher prevalence of PCR positivity. Using seroprevalence data, a sample size of 6-9 animals per population was projected to be adequate for identifying seropositive populations. The presence of I. scapularis was significantly associated with A. phagocytophilum antibodies in WTD. Collectively, the results of this study demonstrate that WTD would be suitable natural sentinels for this emerging zoonotic pathogen.

Age Distribution↗
PubMed2006

Serosurveillance for Anaplasma phagocytophilum antibodies in white-tailed deer (Odocoileus virginianus) in Iowa, USA.

Cases of human granulocytic anaplasmosis have increased in number and are being identified in new geographic areas since its discovery in 1994. White-tailed deer (WTD) become infected with the causative agent, Anaplasma phagocytophilum, and serve as natural sentinels for this organism. In order to determine if A. phagocytophilum is present in the state of Iowa, sera collected from 628 WTD in 2004 from 13 sites and from 282 WTD in 1999 from a single, common site were tested by enzyme-linked immunosorbent assay and Western immunoblotting. A seroprevalence of 9.1% was found among the 2004 samples, and there was no change in seropositivity rates from 1999 to 2004 at the single, common site. As A. phagocytophilum is another tick-borne human pathogen to be identified in the state of Iowa, this study has important implications for health care providers.

Anaplasma phagocytophilum↗
PubMed2004

The prevalence of serum antibodies to tick-borne infections in Mbale District, Uganda: the effect of agro-ecological zone, grazing management and age of cattle.

Between August and October 2000, a cross-sectional study was conducted in smallholder dairy farms in Mbale District, Uganda to assess the prevalence of ticks and tick-borne diseases under different grazing systems and agro-ecological zones and understand the circumstances under which farmers operated. A questionnaire was administered to obtain information on dairy farm circumstances and practices. A total of 102 farms were visited and sera and ticks were collected from 478 animals. Sero-prevalence of tick-borne diseases was determined using an enzyme-linked immunoassay. Acaricides were used indiscriminately but the intensity of their use varied with the grazing system and zone. Cattle from different farms mixed for various reasons. During the dry seasons farmers have to get additional fodder from outside their farms that can result in importation of ticks. The prevalence of ticks and serum antibodies to tick-borne infections differed across the grazing systems and zones. The highest serum antibody prevalence (>60%) was recorded in the lowland zone under the free range and tethering grazing systems. The lowest tick challenge and serum antibody levels (<50%) were recorded in the midland and upland zones under a zero-grazing system. These findings suggest that endemic stability to East Coast Fever, babesiosis and anaplasmosis is most likely to have existed in the lowland zone, particularly, under the tethering and free-range grazing systems. Also, endemic stability for babesiosis existed in the upland zones. Endemic instability for East Coast Fever existed in the midland and upland zones. These structured observational studies are instrumental in planning of control strategies for ticks and tick borne diseases since production systems and the cattle population at high risk of the diseases in the district have been identified.

Aging↗
PubMed1996

Ixodes (Ixodes) scapularis (Acari:Ixodidae): redescription of all active stages, distribution, hosts, geographical variation, and medical and veterinary importance.

The blacklegged tick, Ixodes (Ixodes) scapularis Say, 1821, is redescribed, based on laboratory reared specimens originating in Bulloch County, Georgia. Information on distribution, host associations, morphological variation, and medical/veterinary importance is also presented. A great deal of recent work has focused on this species because it is the principal vector of the agent of Lyme disease (Borrelia burgdorferi Johnson, Schmidt, Hyde, Steigerwaldt & Brenner) in eastern North America. Its distribution appears to be expanding, and includes the state of Florida in the southeastern United States north to the provinces of Nova Scotia and Prince Edward Island, Canada, west to North and South Dakota, United States, and south to the state of Coahuila, Mexico. Although I. scapularis feeds on at least 125 species of North American vertebrates (54 mammalian, 57 avian, and 14 lizard species), analysis of the U.S. National Tick Collection holdings show that white-tailed deer, Odocoileus virginianus (Zimmermann), cattle, Bos taurus L., dogs, Canis lupus L., and other medium-to-large sized mammals are important hosts for adults as are native mice and other small mammals, certain ground-frequenting birds, skinks, and glass lizards for nymphs and larvae. This tick is a polytypic species exhibiting north-south and east-west morphological clines. Analysis of variance and Student-Newman-Keuls multiple comparisons revealed significant interpopulational variation that is expressed most significantly in the nymphal stage. Nymphs from northern (Minnesota, Massachusetts, Maryland) populations had relatively larger basis capituli with shorter cornua (except Maryland) than southern (North Carolina, Georgia) populations. Midwestern populations (Minnesota, Missouri) differed from eastern populations (Massachusetts, Maryland, North Carolina, Georgia) in idiosomal characters (broader scuta, larger coxae III, and IV). In addition to Lyme disease, this tick is also a primary vector of the agent of human and rodent babesiosis, Babesia microti Franca. Under laboratory conditions it has transmitted the agents of deer babesiosis, Babesia odocoilei Emerson & Wright, tularemia, Francisella tularensis McCoy & Chapin, and anaplasmosis, Anaplasma marginale Theiler. Moreover, I. scapularis can reach pest proportions on livestock, and females can cause tick paralysis in dogs.

Animals↗
PubMed2005

Western gray squirrel (Rodentia: Sciuridae): a primary reservoir host of Borrelia burgdorferi in Californian oak woodlands?

In California, dense woodlands have been recognized as important biotopes where humans are exposed to the nymphal stage of the western blacklegged tick, Ixodes pacificus Cooley & Kohls, the primary vector of the Lyme disease spirochete Borrelia burgdorferi sensu stricto (s.s.), in the far-western United States. To identify the principal reservoir host(s) of this spirochete, and of closely related spirochetes in the B. burgdorferi sensu lato (s.l.) complex, in dense woodlands in Mendocino County, California, approximately 50 species of birds and mammals, including wood rats and kangaroo rats, were evaluated as potential hosts for vector ticks and borreliae in 2002 and 2003. Although polymerase chain reaction (PCR) and sequencing analyses revealed that many vertebrate species had been exposed to one or more members of the B. burgdorferi s.l. spirochetal complex, only the western gray squirrel, Sciurus griseus, fulfilled the major criteria for a reservoir host of B. burgdorferi s.s. Ear-punch biopsies from eight of 10 squirrels collected from five separate woodlands were PCR-positive for B. burgdorferi s.s., 47% of I. pacificus larvae (n = 64) and 31% of nymphs (n = 49) removed from squirrels contained B. burgdorferi s.l., and the engorgement status of I. pacificus larvae was associated positively with acquisition of spirochetes. Overall, 83 and 100% of the amplicons sequenced from PCR-positive I. pacificus larvae and nymphs, respectively, were identified as B. burgdorferi s.s, Among the five remaining positive I. pacificus larvae, three contained B. bissettii and two had uncharacterized B. burgdorferi s.l. Borrelia burgdorferi s.s. was detected in one of five larvae and zero of two nymphs of the Pacific Coast tick, Dermacentor occidentalis Marx, that likewise had been removed from squirrels. The rickettsial agent of human anaplasmosis, Anaplasma phagocytophilum, was detected in the blood or ear biopsies of two squirrels and in one (1.6%) of 64 I. pacificus larvae and two (4.1%) of 49 nymphs obtained from squirrels. The one rickettsial-positive larva was coinfected with B. burgdorferi s.s. The apparently high reservoir potential of S. griseus for B. burgdorferi s.s., plus the fact that the geographic distribution of this squirrel coincides well with that of most reported human cases of Lyme disease in this region, indicated that it may be essential for maintaining foci of B. burgdorferi s.s. in certain types of woodlands. The findings with respect to A. phagocytophilum, although of less certain significance, suggest that S. griseus could serve as a secondary host of this rickettsia.

Animals↗
PubMed1996

An assessment of host resistance to ticks on cross-bred cattle in Burundi.

Groups of cattle of four different cross-breeds (Ankole x Friesian, Ankole x Brown-Swiss, Ankole x Guernsey, Ankole x Sahiwal) plus a group of pure Ankole cattle were immunized against tick-borne diseases: East Coast Fever, using a Theileria parva trivalent vaccine, and anaplasmosis, babesiosis and, subsequently, heartwater. With the exception of two small subgroups of Ankole and Ankole x Sahiwal which were treated by weekly spraying using Delnav, all the groups were exposed to natural field challenge of ticks to quantify the development of host resistance to ticks. The results indicate resistance to the most abundant species, Rhipicephalus appendiculatus, as follows: Ankole > Ankole x Sahiwal > Ankole x Brown Swiss > Ankole x Friesian > Ankole x Guernsey. In both Amblyomma variegatum and Rhipicephalus evertsi evertsi, the manifestation of different levels of resistance between breeds is inapparent. Correlations between total ticks and standard female ticks were good for R.appendiculatus (r = 0.73) and R.evertsi (r = 0.51) but poor for A.variegatum (r = 0.31). Correlations between species were reasonably consistent (r = 0.43-0.59). There was no significant correlation between tick burdens and daily liveweight gain (DLWG) over the whole period of the study. Of the cross-bred groups, Ankole x Sahiwal, which had the highest level of resistance, had the highest DLWG. In contrast, the Ankole x Guernsey which developed the lowest level of resistance had only marginally lower DLWG, the difference being only 8 g/day. During the first period of exposure to ticks, however, the untreated Ankole x Sahiwal group had markedly lower DLWG, which may indicate that the development and manifestation of resistance is at the expense of productivity.

Animals↗
PubMed2004

Treatment and prevention of rickettsial and ehrlichial infections.

Rickettsial and ehrlichial infections are both carried by arthropod vectors. Both Rickettsia and Ehrlichia are small intracellular gram-negative coccobacilli. Clinical manifestations of Rickettsia range from spotted fevers to various forms of typhus. Human ehrlichiosis can present as monocytic ehrlichiosis or granulocytic anaplasmosis. Prevention is by avoidance of the responsible vectors. Therapy is usually with doxycycline, but chloramphenicol can also be used.

Animals↗
PubMed2004

Anaplasma phagocytophilum AnkA binds to granulocyte DNA and nuclear proteins.

Human granulocytic anaplasmosis (HGA) is caused by the obligate intracellular bacterium Anaplasma phagocytophilum. The bacterium infects, survives, propagates in, and alters neutrophil phenotype, indicating unique survival mechanisms. AnkA is the only known A. phagocytophilum component that gains access beyond neutrophil vacuoles and is transported to the infected host cell nucleus. The ability of native and recombinant AnkA to bind DNA and nuclear proteins from host HL-60 cells was assessed by the use of immunoprecipitation after cis-diamminedichloroplatinum (cis-DDP) DNA-protein crosslinking, by probing uninfected HL-60 cell nuclear lysates for AnkA binding, and by recovery and sequence analysis of immunoprecipitated DNA. AnkA binds HL-60 cell DNA as well as nuclear proteins of approximately 86, 53 and 25 kDa, whereas recombinant A. phagocytophilum Msp2 or control proteins do not. DNA immunoprecipitation reveals AnkA binding to a variety of target genes in the human genome, including genes that encode proteins with ATPase, tyrosine phosphatase and NADH dehydrogenase-like functions. These data indicate that AnkA could exert some effect on cells through binding to protein:DNA complexes in neutrophil nuclei. Whether AnkA binding leads to neutrophil functional alterations, and how such alterations might occur will depend upon definitive identification of binding partners and associated metabolic and biochemical pathways.

Adenosine Triphosphatases↗
PubMed2005

Anaplasma phagocytophilum inhibits human neutrophil apoptosis via upregulation of bfl-1, maintenance of mitochondrial membrane potential and prevention of caspase 3 activation.

The inhibition of neutrophil apoptosis plays a central role in human granulocytic anaplasmosis. Intracellular signalling pathways through which the obligatory intracellular bacterium Anaplasma phagocytophilum inhibits the spontaneous apoptosis of human peripheral blood neutrophils were investigated. bfl-1 mRNA levels in uninfected neutrophils after 12 h in culture were reduced to approximately 5-25% of 0 h levels, but remained high in infected neutrophils. The eukaryotic RNA synthesis inhibitor, actinomycin D, prevented the maintenance of bfl-1 mRNA levels by A. phagocytophilum. Differences in mcl-1, bax, bcl-w, bad or bak mRNA levels in infected versus uninfected neutrophils were not remarkable. By using mitochondrial fluorescent dyes, Mitotracker Red and JC-1, it was found that most uninfected neutrophils lost mitochondrial membrane potential after 10-12 h incubation, whereas A. phagocytophilum-infected neutrophils maintained high membrane potential. Caspase 3 activity and the degree of apoptosis were lower in dose-dependent manner in A. phagocytophilum-infected neutrophils at 16 h post infection, as compared to uninfected neutrophils. Anti-active caspase 3 antibody labelling showed less positively stained population in infected neutrophils compared to those in uninfected neutrophils after 12 h incubation. These results suggest that A. phagocytophilum inhibits human neutrophil apoptosis via transcriptional upregulation of bfl-1 and inhibition of mitochondria-mediated activation of caspase 3.

Anaplasma phagocytophilum↗