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PubMed1986

Observations on histamine, serotonin, cortisol and cortisol binding globulins in bovine anaplasmosis.

Changes in histamine, serotonin, cortisol and cortisol binding globulin (CBG) levels were observed in Anaplasma marginale-infected bovine calves. In addition, packed cell volume (PCV), platelet number and percentage parasitaemia were also recorded. The whole blood histamine and serotonin values rose significantly (P less than 0.01 and P less than 0.025, respectively) during the acute stage of Anaplasma infection. Higher serum cortisol and CBG levels (P less than 0.05) were observed in acute and carrier infections, respectively. A sharp drop in thrombocyte count (59%) and PCV (33%) was also noticed in clinical anaplasmosis. The results suggest that the higher levels of biogenic amines which are known to produce increased vasodilation, capillary permeability and tissue anoxia and hypercortisolaemia to protect animals from stress and cell damage may play a similar role in the pathogenesis of acute anaplasmosis.

Anaplasmosis↗

PubMed1989

Enzymes of oxidant defence system of leucocytes and erythrocytes in bovine anaplasmosis.

The glutathione oxidant defence system in leucocytes and erythrocytes of six Anaplasma marginale-infected calves was examined by assaying glutathione peroxidase (GSH-px), glutathione reductase (GSSG-R), reduced glutathione (GSH) and superoxide dismutase (SOD). In addition, GSH-S-transferase and arginase levels were measured in leucocytes. There was a significant decline in the activities of leucocyte GSH-px, GSSG-R and SOD in the post-patent period; whereas arginase activity rose significantly following the patent period of anaplasmosis. The activity of erythrocyte SOD declined in the post-patent stage. The infection also caused a decline in red cell GSH (P less than 0.05). The results suggest that the glutathione oxidant defence system of peripheral blood cells is significantly influenced by the Anaplasma infection.

Anaplasmosis↗

PubMed1995

Recent developments in elucidating tick vector relationships for anaplasmosis and equine piroplasmosis.

This brief review focuses first on several epidemiologically relevant aspects of anaplasmosis, including: (1) the role of male ticks as intrastadial, biological vectors of Anaplasma through interhost transfer; (2) the application of molecular diagnostic assays in assessing tick vector competence and evaluating the role of chronically infected carrier cattle as sources of Anaplasma marginale infection in vector ticks; (3) opportunities provided by a recently developed in vitro tick feeding system in quantitating studies of tick-hemoparasite-host interactions. Lastly, current knowledge of the status of New World ticks as experimental and/or natural vectors of equine piroplasms is discussed.

Anaplasmosis↗

PubMed1995

Targeting ticks for control of selected hemoparasitic diseases of cattle.

Development in and transmission of hemoparasites by tick vectors are phenomena closely synchronized with the tick feeding cycle. In all known life cycles, initial infection of tick tissues occurs in midgut epithelial cells and transmission is effected as ticks feed after parasites have developed and multiplied in salivary glands. Many factors reviewed affect development and transmission of hemoparasites by ticks including age of ticks, artificial temperature, climate and/or season, tick stage or sex, hemoparasite variation, concurrent infection of ticks with other pathogens, host cell susceptibility, transovarial transmission, effect of hemoparasites on tick biology, and the effect of infecting parasitemia level in cattle on infection rates in ticks. Four hemoparasites of cattle, Anaplasma marginale, Cowdria ruminantium, Theileria parva, and Babesia spp., are all dependent on ticks for biological transmission. Babesia is transmitted transovarially whereas the other three are transmitted transstadially. Mechanical transfer of infective blood via fomites and mouthparts of biting arthropods is also a major means of transmission for Anaplasma marginale but not of the others. Potential control methods for hemoparasites that target parasites as they are developing in their respective tick hosts include tick control, vaccines (against ticks and parasites), and drugs (against ticks and parasites). Successful application of control strategies will be dependent upon thorough understanding of parasite developmental cycles, biology of the tick vectors and the immune response of cattle to ticks and to hemoparasites. The most effective control measures will be those that are targeted against both ticks and the hemoparasites they vector.

Animals↗

PubMed2006

Etiology of tick-borne febrile illnesses in adult residents of North-Eastern Poland: report from a prospective clinical study.

Febrile tick-borne diseases can be caused by different pathogens. The study objective was to prospectively determine the etiology of infection among adults exposed to tick bite and to detect acute human granulocytic anaplasmosis (HGA), a recently emerging infection, in north-eastern Poland. We evaluated 68 patients (34 males and 34 females, mean age 44.2+/-15 years) who had fever within 4 weeks after a tick bite. Paired sera were tested for antibodies against tick-borne encephalitis virus, Borrelia burgdorferi and Anaplasma phagocytophilum. Microscopic examination of EDTA whole blood and nested PCR targeting A. phagocytophilum 16S rDNA gene fragment were carried out at enrolment. Tick-borne infections were diagnosed in 57 individuals (84%). Sixty-four patients were hospitalised. Tick-borne encephalitis (TBE) was revealed in 49 cases; in 39 patients as a single infection, in three patients concurrent with erythema migrans or other Lyme borreliosis symptoms, in five patients concurrent with probable or possible Lyme borreliosis, and in two patients concurrent with A. phagocytophilum infection. Diagnosis of HGA was confirmed in one patient by PCR and in another one by seroconversion. In two further individuals, A. phagocytophilum infection was confirmed by immunofluorescence (antibody titers > or = 1:128), which, however, does not fulfil the ESCAR (European Society of Clinical Microbiology and Infectious Diseases Study Group on Coxiella, Anaplasma, Rickettsia, and Bartonella) criteria for HGA case definition. Lyme borreliosis was diagnosed in 16 patients; in six of them as a single infection. Both confirmed HGA cases and seropositive individuals had A. phagocytophilum infection concurrent either with Lyme borreliosis (two cases) or with TBE (two cases). The clinical course was severe in three and moderate to mild in the remaining TBE cases, as well as in all Lyme borreliosis and HGA cases. TBE was found the most prevalent disease among adults with febrile illnesses occurring after a tick bite in north-eastern Poland. Concurrent tick-borne infections were frequent with multiple pathogens involved. Two confirmed acute HGA cases (fulfilling the European case definition criteria defined by ESCAR) were detected for the first time in a prospective manner in Poland.

Adolescent↗

PubMed2001

Seroprevalences of vector-transmitted infections of small-holder dairy cattle in coastal Kenya.

A cross-sectional study was carried out from July to September 1989 in Kaloleni Division, Coast Province, Kenya to estimate the prevalence of vector-transmitted diseases in small-holder dairy cattle and to identify the risk factors associated with different management systems. One hundred and thirty of the 157 herds with dairy cattle in Kaloleni Division were surveyed. These were from three agro-ecological zones (coconut-cassava, cashew nut-cassava and livestock-millet), comprised two management systems (stall-feeding and herded grazing) and were herds with either dairy cattle only or with Zebu and dairy cattle. A formal questionnaire sought answers to questions on cattle health and management practices. A total of 734 dairy and 205 Zebu cattle in 78 dairy and 52 mixed (dairy and Zebu) herds were sampled and screened for haemoparasites (Trypanosoma, Anaplasma, Babesia, and Theileria infections). Sera were tested for antibodies to Theileria parva, using the schizonts-antigen indirect fluorescent-antibody (IFA) test and to antibodies for Babesia bigemina and antigens to Anaplasma marginale by enzyme-linked immunosorbent assay (ELISA). Packed-cell volume (PCV) also was measured. Tick-control measures were practised by all except three of the farmers. Despite this, overall seroprevalence to T. parva was >70%--suggesting either that control practices were not strictly implemented or they were ineffective. The seroprevalence of T. parva in adult cattle kept in stall-feeding systems in the coconut-cassava zone was significantly lower (57+/-8% (S.E.)) than in herded-grazing systems (79+/-3%) and there was no association between antibody prevalence and age of cattle in this zone. Antibody prevalences in cattle in the cashew nut-cassava and the drier livestock-millet zone increased with age. Cattle in herded-grazing systems had an overall lower seroprevalence of T. parva infection in the livestock-millet zone (45+/-6%) than in the other two zones. Analysis was confined to the coconut-cassava zone for B. bigemina and to the coconut-cassava and cashew nut-cassava zones for A. marginale. Mean prevalences of B. bigemina were 40.9+/-9 and 73+/-6% for dairy cattle under stall-feeding and herded-grazing systems, respectively, and increased with age. Antigen prevalences of A. marginale were over 80% in all age groups of cattle in the coconut-cassava and cashew nut-cassava zones. Overall trypanosome prevalence in cattle was <1%. Trypanocidal treatment was uncommon. The variations in antibody prevalence associated with risk factors such as feeding system, agro-ecological zone and age of animal suggest that management system influenced exposure to tick-borne infection (particularly, T. parva infections) in small-holder dairy cattle in coastal Kenya.

Agriculture↗

PubMed2005

Detection of medically important Ehrlichia by quantitative multicolor TaqMan real-time polymerase chain reaction of the dsb gene.

Ehrlichia species are the etiological agents of emerging and life-threatening tick-borne human zoonoses, in addition to causing serious and fatal infections in companion animals and livestock. We developed the first tricolor TaqMan real-time polymerase chain reaction assay capable of simultaneously detecting and discriminating medically important ehrlichiae in a single reaction. Analytical sensitivity of 50 copies per reaction was attained with templates from Ehrlichia chaffeensis, Ehrlichia ewingii, and Ehrlichia canis by amplifying the genus-specific disulfide bond formation protein gene (dsb). Ehrlichia genus-specific dsb primers amplified DNA from all known Ehrlichia species but not from other rickettsial organisms including Anaplasma platys, Anaplasma phagocytophilum, Rickettsia conorii, or Rickettsia typhi. High species specificity was attained as each species-specific TaqMan probe (E. chaffeensis, E. ewingii, and E. canis) identified homologous templates but did not cross-hybridize with heterologous Ehrlichia templates at concentrations as high as 10(8) copies. Identification of E. chaffeensis, E. ewingii, and E. canis from natural and experimental infections, previously confirmed by polymerase chain reaction and serological or microscopic evidence, demonstrated the comparable specificity and sensitivity of the dsb real-time assay. This assay provides a powerful tool for prospective medical diagnosis for human and canine ehrlichioses and for ecologic and epidemiological studies involving arthropod and mammalian hosts.

Animals↗

PubMed2005

Immunity to the ehrlichiae: new tools and recent developments.

PURPOSE OF REVIEW: Discusses recent developments in the study of immunity and host defense against the monocytic ehrlichiae in 2003 and 2004. The review does not address anaplasmoses, as the anaplasmae were recently re-classified into the genus Anaplasma, and are distinct in cell tropism from the ehrlichiae. RECENT FINDINGS: The features of the immune responses against these emerging Gram-negative obligate intracellular pathogens are only beginning to be understood. Important advances in our ability to study host defense include the development of new experimental mouse models. Recent studies have defined possible mechanisms of innate immune subversion in human monocytes, as well as roles for lymphocyte subsets and type I cytokines during mouse infection. Other studies in the mouse suggest that cytokine production by CD8 T cells may contribute to immunopathology. New data also support a role for humoral immunity during host defense against these intracellular pathogens. SUMMARY: The use of new animal models will facilitate research of the mechanisms of innate, adaptive, and pathological immune responses, and will enhance our understanding of human immunity to the ehrlichiae as well as to other pathogenic intracellular bacteria.

Animals↗

PubMed1980

A comparison of direct fluorescent antibody and Giemsa staining for the post-mortem diagnosis of anaplasmosis.

Direct fluorescent antibody (DFA) and Giemsa staining of Anaplasma marginale were compared in smears collected serially at post-mortem (PM) from 11 experimentally infected calves. Once smears had been prepared and air-dried they could be held for at least 5 days before staining with either technique with no noticeable change in staining quality. DFA staining was more sensitive in detecting anaplasms in smears than Giemsa staining. Anaplasma spp could be differentiated from Babesia bovis and B. bigemina by DFA staining but there were cross reactions between A. marginale and A. centrale. Blood smears prepared from subcutaneous vessels in the legs provided better diagnostic material than kidney, heart and lung smears. Brain smears were not suitable for PM diagnosis using either staining technique.

Anaplasmosis↗

PubMed1991

16S ribosomal DNA amplification for phylogenetic study.

A set of oligonucleotide primers capable of initiating enzymatic amplification (polymerase chain reaction) on a phylogenetically and taxonomically wide range of bacteria is described along with methods for their use and examples. One pair of primers is capable of amplifying nearly full-length 16S ribosomal DNA (rDNA) from many bacterial genera; the additional primers are useful for various exceptional sequences. Methods for purification of amplified material, direct sequencing, cloning, sequencing, and transcription are outlined. An obligate intracellular parasite of bovine erythrocytes, Anaplasma marginale, is used as an example; its 16S rDNA was amplified, cloned, sequenced, and phylogenetically placed. Anaplasmas are related to the genera Rickettsia and Ehrlichia. In addition, 16S rDNAs from several species were readily amplified from material found in lyophilized ampoules from the American Type Culture Collection. By use of this method, the phylogenetic study of extremely fastidious or highly pathogenic bacterial species can be carried out without the need to culture them. In theory, any gene segment for which polymerase chain reaction primer design is possible can be derived from a readily obtainable lyophilized bacterial culture.

Bacteria↗

PubMed1995

Detection of Theileria annulata in blood samples of carrier cattle by PCR.

We report the detection of Theileria annulata, the causative agent of tropical theileriosis, by PCR in blood samples obtained from carrier cattle. The assay employs primers specific for the gene encoding the 30-kDa major merozoite surface antigen of T. annulata. A 721-bp fragment was amplified from blood samples taken monthly from calves experimentally infected with one of four different stocks of T. annulata originating in either Mauritania, Portugal, Spain, or Turkey. At the end of the experiment, five animals carried the infection for 12 months and two animals remained infected for 15 months. DNAs from six other Theileria species, T. parva, T. mutans, T. sergenti, T. buffeli, T. velifera, and T. taurotragi, were not amplified. Moreover, DNAs from four other hemoparasites (Anaplasma centrale, Anaplasma marginale, Babesia bovis, and Babesia bigemina) were also not amplified. As a control, primers derived from the small subunit rRNA gene of Theileria spp. amplified a 1.1-kb DNA fragment from all Theileria species examined but not from the other four hemoparasites. As few as two to three parasites per microliter of infected blood in a 50-microliters sample volume were detected by Southern or microplate hybridization with a T. annulata-specific cDNA probe. In addition, 92 field samples obtained from cattle in Spain were tested; 22% were positive in blood smears, 40% were positive by immunofluorescent antibody test, and 75% were positive for T. annulata by PCR. The method provides a useful diagnostic tool for detecting T. annulata carrier cattle.

Animals↗

PubMed2001

Citrate synthase gene sequence: a new tool for phylogenetic analysis and identification of Ehrlichia.

The sequence of the citrate synthase gene (gltA) of 13 ehrlichial species (Ehrlichia chaffeensis, Ehrlichia canis, Ehrlichia muris, an Ehrlichia species recently detected from Ixodes ovatus, Cowdria ruminantium, Ehrlichia phagocytophila, Ehrlichia equi, the human granulocytic ehrlichiosis [HGE] agent, Anaplasma marginale, Anaplasma centrale, Ehrlichia sennetsu, Ehrlichia risticii, and Neorickettsia helminthoeca) have been determined by degenerate PCR and the Genome Walker method. The ehrlichial gltA genes are 1,197 bp (E. sennetsu and E. risticii) to 1,254 bp (A. marginale and A. centrale) long, and GC contents of the gene vary from 30.5% (Ehrlichia sp. detected from I. ovatus) to 51.0% (A. centrale). The percent identities of the gltA nucleotide sequences among ehrlichial species were 49.7% (E. risticii versus A. centrale) to 99.8% (HGE agent versus E. equi). The percent identities of deduced amino acid sequences were 44.4% (E. sennetsu versus E. muris) to 99.5% (HGE agent versus E. equi), whereas the homology range of 16S rRNA genes was 83.5% (E. risticii versus the Ehrlichia sp. detected from I. ovatus) to 99.9% (HGE agent, E. equi, and E. phagocytophila). The architecture of the phylogenetic trees constructed by gltA nucleotide sequences or amino acid sequences was similar to that derived from the 16S rRNA gene sequences but showed more-significant bootstrap values. Based upon the alignment analysis of the ehrlichial gltA sequences, two sets of primers were designed to amplify tick-borne Ehrlichia and Neorickettsia genogroup Ehrlichia (N. helminthoeca, E. sennetsu, and E. risticii), respectively. Tick-borne Ehrlichia species were specifically identified by restriction fragment length polymorphism (RFLP) patterns of AcsI and XhoI with the exception of E. muris and the very closely related ehrlichia derived from I. ovatus for which sequence analysis of the PCR product is needed. Similarly, Neorickettsia genogroup Ehrlichia species were specifically identified by RFLP patterns of RcaI digestion. If confirmed this technique will be useful in rapidly identifying Ehrlichia spp.

Animals↗

PubMed2006

New findings on members of the family Anaplasmataceae of veterinary importance.

Members of the family Anaplasmataceae are obligate intracellular Gram-negative bacteria that naturally infect a variety of wild and domestic animal species, the spillover of which may lead to zoonosis. I discuss new findings on members of the family Anaplasmataceae of veterinary importance and therefore, I will describe the recent findings on Neorickettsia risticii in the trematode and related Neorickettsia species. I also will review the recent progress on Aegyptianella pullorum and other Aegyptianella sp., "Candidatus Neoehrlichia mikurensis" and Anaplasma phagocytophilum strains in various hosts. The whole genome sequences of two important veterinary pathogens-Anaplasma marginale, the bovine anaplasmosis agent, and Ehrlichia (formerly Cowdria) ruminantium, the agent of heartwater of ruminants-have been published. Taken together, these advances in research of the family Anaplasmataceae in the veterinary field provide us with insights into the evolution, reservoir, and transmission of these organisms in nature and their pathogenesis in natural and accidental hosts. It is through this work that surveillance, diagnosis, preventive measures, and treatment of ehrlichioses of both animals and humans can be improved.

Anaplasmataceae↗

PubMed2006

Comparative genomics of emerging human ehrlichiosis agents.

Anaplasma (formerly Ehrlichia) phagocytophilum, Ehrlichia chaffeensis, and Neorickettsia (formerly Ehrlichia) sennetsu are intracellular vector-borne pathogens that cause human ehrlichiosis, an emerging infectious disease. We present the complete genome sequences of these organisms along with comparisons to other organisms in the Rickettsiales order. Ehrlichia spp. and Anaplasma spp. display a unique large expansion of immunodominant outer membrane proteins facilitating antigenic variation. All Rickettsiales have a diminished ability to synthesize amino acids compared to their closest free-living relatives. Unlike members of the Rickettsiaceae family, these pathogenic Anaplasmataceae are capable of making all major vitamins, cofactors, and nucleotides, which could confer a beneficial role in the invertebrate vector or the vertebrate host. Further analysis identified proteins potentially involved in vacuole confinement of the Anaplasmataceae, a life cycle involving a hematophagous vector, vertebrate pathogenesis, human pathogenesis, and lack of transovarial transmission. These discoveries provide significant insights into the biology of these obligate intracellular pathogens.

Animals↗

PubMedDate not supplied

Chronic canine ehrlichiosis (Ehrlichia canis): a retrospective study of 19 natural cases.

Nineteen dogs from Greece with chronic ehrlichiosis were studied. The dogs exhibited bicytopenia or pancytopenia, bone marrow hypoplasia, seroreactivity to Ehrlichia canis (E. canis) antigens, and had no history of drug or radiation exposure. Anorexia, depression, severe bleeding tendencies, hypoalbuminemia, and increased serum alanine aminotransferase activity were also hallmarks of the disease. All these animals eventually died, irrespective of the treatment applied. Some dogs were also serologically positive for Rickettsia conorii, Leishmania infantum (L. infantum), and Bartonella vinsonii subspp. berkhoffii. Polymerase chain reaction testing of bone marrow samples revealed E. canis, Anaplasma phagocytophilia, Anaplasma platys, and L. infantum in some dogs. Concurrent infections did not appear to substantially influence the clinical course and final outcome of the chronic canine ehrlichiosis.

Animals↗

PubMed2001

Health protocol for translocation of free-ranging elk.

When considering an elk (Cervus elaphus) restoration program, wildlife managers must evaluate the positive and negative elements of translocation. We prepared this protocol to give an overview of health considerations associated with translocation of elk, with an emphasis on movement of free-ranging elk from western North America to the southeastern USA. We evaluated infectious agents and ectoparasites reported in elk from two perspectives. First, we made a qualitative estimate of the ability of the agent to be introduced and to become established. This was done using a selected set of epidemiologic factors. Second, if there was a good possibility that the organism could become established in the release area, the potential pathological consequences for elk and other wildlife, domestic animals, and humans were assessed via examination of the literature and consultation with other animal health specialists. The results of these evaluations were used to classify infectious agents and ectoparasites as low risk (n = 174), unknown risk (n = 10), and high risk (n = 9). We classified Anaplasma marginale, Anaplasma ovis, Mycobacterium paratuberculosis, Pasteurella multocida serotype 3, Elaphostrongylus cervi, Dicrocoelium dendriticum, Fascioloides magna, Echinococcus granulosus, Dermacentor albipictus, and Otobius megnini as unknown risks. High risk infectious agents and ectoparasites were the agent of chronic wasting disease, Brucella abortus, Mycobacterium bovis, Dermacentor andersoni, Ixodes pacificus, and Psoroptes sp. Parelaphostrongylus tennis, Elaeophora schneideri, and a Babesia sp. are parasites endemic in the southeastern USA that may present a "reverse risk" and adversely affect elk if released in some parts of the region. We developed a five-component protocol to reduce the risk of introduction of high risk infectious agents and ectoparasites that included: (1) evaluation of the health status of source populations, (2) quarantines, (3) physical examination and diagnostic testing, (4) restrictions on translocation of animals from certain geographic areas or populations, and (5) prophylactic treatment.

Animal Welfare↗

PubMed1976

Survey of anaplasmosis in elk of the Clearwater National Forest (Idaho).

Samples of blood from 27 free-roaming elk (Cervus canadensis canadensis) from the Clearwater National Forest in north central Idaho were tested by the rapid card agglutination test and complement-fixation test for the presence of antibodies against Anaplasma marginale. The serum card test and complement-fixation test gave incomplete and false-positive reactions; the plasma card test did not give any reactions. Anaplasma bodies or other blood parasites were not observed in stained smears of elk blood. Blood from 11 elk, including 2 that were serum card test-positive, did not produce clinical, hematologic, or serologic evidence of infection in 3 anaplasmosis-susceptible bovine calves.

Anaplasmosis↗

PubMed2004

[Ehrlichiosis--a disease rarely recognized in Poland].

Ehrlichioses constitute a group of acute zoonoses caused by the infection with the microorganisms belonging to the genera Ehrlichia, Anaplasma and Neorickettsia (Rickettsiaceae). Presence of human granulocytic ehrlichiosis (HGE) caused by infection with Anaplasma phagocytophila (A. phagocytophila) and transmitted by Ixodes ricinus tick has been confirmed in Poland. All the cases described so far were noted in the area of endemic prevalence of Lyme borreliosis (north-east of Poland). Lack of characteristic symptoms makes the diagnosis of HGE difficult. It should be suspected in patients exposed to tick bites during the preceding few weeks, in whom acute, febrile illness accompanied by leucopenia and thrombocytopenia develops. The course of infection may be serious, or even life-threatening, in patients more than 40 years old; consequences of co-infection with A. phagocytophila and other tick-borne pathogens remain not well known. There is probably no risk of the development of chronic HGE. As specific diagnostic methods are not widely accessible in Poland, empirical therapy with doxycycline may be of benefit in suspected HGE cases.

Diagnosis, Differential↗