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Tick-borne diseases of cattle in Paraguay. I. Seroepidemiological studies on anaplasmosis and babesiosis.

A serological survey was carried out in Paraguay to determine the prevalence of antibodies to Anaplasma marginale, Babesia bigemina and Babesia bovis. The results suggest that the parasites are widely distributed throughout the country with prevalence rates of 92% for A. marginale, 79% for B. bigemina and 71% for B. bovis. Epidemiological studies on calves demonstrated that initial infection with the three blood parasites occurs in most animals within the first nine months of life. It is likely therefore that most of the country can be considered as being enzootically stable with respect to anaplasmosis and babesiosis.

Anaplasma↗

Field observations on adult zebu bulls immunised against anaplasmosis by infection and treatment plus integrated tick control.

Fourteen adult zebu bulls which were negative for Anaplasma marginale infection both serologically and on blood smear examinations were infected with a virulent Nigerian isolate of A. marginale. Forty days following the immunising infection when clinical reactions were established groups of seven animals were treated with either two doses of imidocarb dipropionate (2 X 5 mg/kg given intramuscularly 14 days apart) or a single intramuscular dose of long-acting oxytetracycline (5 mg/kg). Following clinical recovery, two weeks after the first treatment the immunised animals and seven susceptible controls were then introduced into a tick-infested area and held there for 15 months with regular tick control. There were significant differences between the mean weight gains of surviving animals in the oxytetracycline group and the controls and surviving animals in the imidocarb group and control cattle. The protective effect of tick control alone was inferior to that of integrated tick control plus anaplasmosis control by either method of immunisation. Losses occurred in all groups due mainly to the effects of other tick-borne diseases, babesiosis and heartwater.

Anaplasma↗

Immunisation of smallholder dairy cattle against anaplasmosis and babesiosis in Malawi.

A field study was conducted in the Southern Region of Malawi to evaluate the possible benefits of immunisation of improved dairy cattle against Anaplasma marginale, Babesia bigemina and Babesia bovis. Friesian crossbred heifers were immunised when they were being reared on Government farms. They were then issued to smallholder farmers, together with unvaccinated controls, where many of them were exposed to heavy tick infestation. Vaccination was shown to provide a significant degree of protection against babesiosis on the smallholder farms; 15/32 unvaccinated controls developed clinical disease as compared to only 3/28 vaccinates.

Anaplasma↗

Vaccines against hemoparasitic diseases in Israel with special reference to quality assurance.

Four vaccines against hemoparasitic diseases (anaplasmosis, babesiosis and theileriosis) and a vaccine against besnoitiosis are currently used in Israel. These vaccines contain live attenuated parasites derived from cell culture (Theileria annulata and Besnoitia besnoiti) or from blood of infected, splenectomized calves (Babesia bigemina, B. bovis and Anaplasma centrale). Cryopreserved master seed is used to initiate production of the vaccines. Quality control performed during the preproduction period is particularly important with blood-derived vaccines. Post production quality control comprises sterility, potency (viability of immunizing organisms), safety (degree of attenuation) and efficacy (capacity to protect against virulent parasite stock). All vaccines are stored and dispatched to the field in a concentrated frozen state. The culture-derived vaccines are safe for all varieties of cattle, regardless of age or physiological condition, whereas the blood-derived vaccines are recommended mainly for young cattle, the age limit varying with the type of vaccine and breed of cattle. The viability of T. annulata infected cells in the anti-theilerial vaccine is tested after thawing by in vitro plating efficiency and the infectivity of blood-derived vaccines is tested by titration in susceptible cattle.

Anaplasma↗

First documented case of human granulocytic ehrlichiosis in Austria.

We report the first documented case of human granulocytic ehrlichiosis (HGE) in Austria. The infection was acquired near Arzl in the surroundings of Innsbruck in northern Tyrol. Except for a biphasic course of illness, presentation in this 33-year-old female patient was comparable to clinical findings observed in other European adults with high fever, transient thrombocytopenia, elevated levels of CRP and LDH, arthralgias, myalgias, fatigue and subfebrile temperature. Flu-like symptoms started about seven days after a tick bite and lasted for ten days, followed by an asymptomatic interval of three days and an acute onset of fever up to 39.5 degrees C on day 20. On admission, the patient showed high antibody titres against Anaplasma phagocytophilum (IgG 1:1024, IgM 1:640); six weeks later the IgG-titre had risen to 1:2048, and IgM-levels had fallen below 1:40. The demonstration of anti-platelet antibodies in acute-phase serum was noteworthy. We conclude that also in Austria HGE should be considered in patients with febrile thrombocytopenia, especially when the medical history reveals recent tick exposure.

Adult↗

Intrahaplotype and interhaplotype pairing of bovine leukocyte antigen DQA and DQB molecules generate functional DQ molecules important for priming CD4(+) T-lymphocyte responses.

Antigen-specific CD4(+) T-lymphocyte responses are restricted by major histocompatibility complex class II molecules, which influence T-cell priming during infection. Human leukocyte antigen (HLA) and bovine leukocyte antigen (BoLA) DRB3 and DQ genes are polymorphic, but unlike HLA, many BoLA haplotypes have duplicated DQ genes, and antibody-blocking studies indicated that BoLA-DQ molecules present various pathogen epitopes. Limited experimentation also suggested that BoLA-DQ molecules formed by interhaplotype pairing of A and B chains are functional. To compare antigen presentation by DR and DQ molecules and to definitively demonstrate functional BoLA-DQ molecules derived from interhaplotype pairing, different combinations of DR or DQ A and B proteins were expressed with CD80 in 293-F cells for use as antigen-presenting cells (APCs). This approach identified 11 unique restriction elements including five DR and six DQ pairs for antigen-specific CD4(+) T-cell responses against tick-transmitted bovine hemoparasites Anaplasma marginale or Babesia bovis. Interhaplotype pairing of DQ A and B molecules was demonstrated. Testing of six expressed DQA/B pairs from an animal with duplicated DQ haplotypes (DH16A/DH22H) demonstrated that an interhaplotype pair, DQA*2206/DQB*1301, presented A. marginale peptide B. In DH22H and DH16A homozygous animals, DQA*2206 was tightly linked with DQB*1402, and DQA*22021 was linked with DQB*1301. APCs from these donors could not present peptide B, confirming that DQA*2206/DQB*1301 encoded a functional interhaplotype pair. Functional BoLA-DQ molecules are generated by both intrahaplotype and interhaplotype pairing of A and B chains and play a similar role to BoLA-DR in priming helper T-cell responses to important pathogens.

Amino Acid Sequence↗

High seroprevalence of granulocytic ehrlichiosis distinguishes sheep that were the source of an alimentary epidemic of tick-borne encephalitis.

A sheep herd from which contaminated cheese was produced, causing 21 cases of alimentary tick-borne encephalitis (TBE) in human beings, was tested serologically for the presence of specific antibodies against both the TBE virus (TBEV) and Anaplasma phagocytophilum, the cause of tick-borne fever (TBF) in ruminants, and compared with three other herds variously exposed to tick bites but without any TBE history. Virus-neutralisation (VN) with the TBEV strain Hypr and CV-1 cells was used in TBE tests, and indirect immunofluorescence assay (IFA) with neutrophils from goats experimentally infected with A. phagocytophilum was used for TBF testing. In 13 sheep from the incriminated herd (N =41), VN titres ranging from 1/4 to 1/128 traced previous TBE infection and all sheep had elevated titres of A. phagocytophilum antibodies ranging from 1/80 to 1/5120 in IFA, whereas two other herds (N = 8 and 9) were seronegative for TBEV and had significantly lower levels of A. phagocytophilum antibodies, corresponding to a lesser challenge from TBF. A control herd (N = 10) that was grazed on tick-free meadows in north Norway was completely seronegative. The respective distributions of positive titres of A. phagocytophilum and TBEV antibodies in the incriminated herd were not mutually random; the animals with higher anti-A. phagocytophilum titres tended to have lower anti-TBEV titres and vice versa (Spearman correlation coeff. =-0.86, p< or =0.01). The authors hypothesize that the immunosuppressive effect of TBF co-infection in sheep could be a contributory cause of TBE-virus contamination of milk, an aspect of TBE epidemiology that has not been considered thus far.

Anaplasma phagocytophilum↗

Epidemiological, clinical and laboratory characteristics of patients with human granulocytic anaplasmosis in Slovenia.

Human granulocytic anaplasmosis (HGA) has been recently recognized as an emerging tick-borne disease. Several reports indicate the presence of infection with Anaplasma phagocytophilum in Europe. Between January 1996 and December 2004, 24 adult patients with proven HGA were identified in a prospective study conducted at the Department of Infectious Diseases, University Medical Center Ljubljana, Slovenia, on the etiology of febrile illnesses occurring within 30 days after a tick bite. The diagnosis of acute HGA was established from seroconversion in 18 (75%) patients or at least a four-fold increase in antibody titers to A. phagocytophilum antigens in six (25%) patients and molecular identification of ehrlichial organisms in 15 (62.5%) patients. Clinical characteristics and laboratory findings were similar to those reported from the other European countries. All the patients had an acute febrile illness with headache, malaise, myalgia and/or arthralgia. Leukopenia was found in 16 (66.7%) patients, thrombocytopenia in 20 (83.3%), abnormal liver function test results in 23 (95.8%), elevated erythrocyte sedimentation rates in 18 (75%), and elevated concentration of C-reactive protein in 23 (95.8%). The disease course was relatively mild; none of the patients died and no long-term sequelae were found during a follow-up of one year even though only 15 (62.5%) were treated with doxycycline. At the examination one year after the first visit, 16/24 (66.7%) patients tested seropositive (> or =1 : 256) for A. phagocytophilum antibody, and two years after the first visit positive titers were still present in 10/18 (55.6%) patients.

Adult↗

Ticks parasitizing wild birds in Portugal: detection of Rickettsia aeschlimannii, R. helvetica and R. massiliae.

From January 2002 to December 2004, 152 ticks were collected from 40 wild birds recovered in Santo André Natural Reserve and Monsanto Forestal Park, Portugal mainland. Five ticks species were identified from 22 species of birds, and new host record were provided for some species. In addition, 32 (21%) ticks were screened by PCR to detect infections with agents belonging to order Rickettsiales: Anaplasma phagocytophilum, Ehrlichia chaffeensis, and Rickettsia spp. PCR amplicons were obtained in 5 (15.6%) tick samples. Rickettsia DNA exhibiting gltA sequences similar to those of Rickettsia aeschilimannii, R. helvetica and R. massiliae were identified in Hyalomma marginatum, Ixodes ventalloi and in Rhipicephalus turanicus, respectively. This is the first report of rickettsiae infections in ticks collected from wild birds in Portugal. Giving the results presented above wild birds play an important role in the maintenance and dissemination of several tick species and associated rickettsiae.

Anaplasma phagocytophilum↗

Clinical and serological evidence of bovine babesiosis and anaplasmosis in St. Lucia.

One hundred fifty-nine Holstein calves were imported into St. Lucia from the U.S.A. An outbreak of babesiosis occurred 17 days post-arrival, and an outbreak of anaplasmosis occurred 5 months after importation. Sera obtained 3, 6, and 12 months post-importation revealed a high prevalence of IFA titres to Babesia bovis and B. Bigemina 3 months after arrival and an increase in titres to Anaplasma marginale 6 months after arrival. Sera obtained from native cattle from several places on the island indicated infection rates of 80, 65 and 64% with A. marginale, B. bigemina and B. bovis, respectively. The rapid card test only indicated a 25% prevalence of infection of native cattle by A. marginale. This low prevalence was probably due to deterioration of serological activity during shipment.

Anaplasma↗

Studies on ticks of veterinary importance in Nigeria. XI. Observations on the biology of ticks detached from the red-flanked duiker (Cephamophys rufulatus) and parasites encountered in their blood.

Some aspects of the biology of Amblyomma variegatum and Boophilus decorolatus detached from red-flanked duikers were studied. Smears were also made from the blood of the duikers and examined for the presence of parasites. Anaplasma marginale was the only blood parasite found in the blood smears. Compared with ticks detached from cattle, duiker ticks produced a greater number of eggs. The lengths and breadths of eggs produced by duiker ticks were greater than those of cattle ticks and unlike A. variegatum detached from cattle, the lengths and breadths of eggs of earlier and later ovipositions of this species detached from duiker were similar. Whereas deformed eggs characterised by circular shape and small size constituted a small percentage of eggs of cattle ticks, none was found among eggs of duiker ticks. The pre oviposition, oviposition and eclosion periods, microscopic egg structure, embryonic development, hatching patterns and mortality rates of the eggs of ticks from both hosts were similar.

Anaplasma↗

Cross-reaction of tick salivary antigens in the Boophilus microplus-cattle system.

Calves were immunized with Boophilus microplus saliva, filtered through Millipore membranes, in Freund's complete adjuvant. Serum samples were tested by passive hemagglutination against Babesia bigemina, Anaplasma marginale, B. microplus larvae extract, Stomoxys calcitrans extract and B. microplus saliva. After immunization, titers to saliva, larval tick-extract and to S. calcitrans were increased. The challenge with live tick larvae enhanced the formation of antibodies against larva extract, fly extract and tick saliva, which supports the idea that under natural controlled conditions this cross-reactivity could occur.

Anaplasma↗

Post-thawing viability of vaccines for bovine babesiosis and anaplasmosis cryopreserved with glycerol.

Live frozen vaccines containing Babesia bovis, Babesia bigemina or Anaplasma centrale were prepared using glycerol as cryoprotectant and stored in liquid nitrogen. The viability of the vaccines was tested inoculating calves 1 h (n = 12), 2 h (n = 12), 12 h (n = 6) and 24 h (n = 6) after thawing. Babesia bovis and A. centrale were detected in thin and/or thick blood smears in all vaccinated calves; however, 1 of 12 calves inoculated 1 h after thawing and 3 of 6 calves inoculated 24 h after thawing did not develop a B. bigemina parasitaemia. The longer post-thawing durability of frozen vaccines cryoprotected with glycerol compared with those cryoprotected with dimethyl sulfoxide, presented by other authors, will extend their use under field conditions.

Anaplasma↗

Recent developments in the molecular biology of anaplasmosis.

Recent applications of DNA analysis, cloning, sequencing and expression technology have resulted in significant advances in our understanding of the hemoparasite Anaplasma marginale. Analysis of 16S ribosomal RNA has confirmed a phylogenetic position close to Ehrlichia sp. and Cowdria ruminantium. Intact genomic DNA of A. marginale digested with SfiI separates into bands from 14 to 170 kbp on pulse-field gels, with a total genome size of 1200-1260 kbp and G + C content of 56 mol%. Major surface proteins (MSP1-MSP5) have been identified and DNA coding sequences are available for most of these. These data have revealed that MSPs may be quite polymorphic between different geographic isolates, may be encoded by multi-gene families, and have some similar features to other prokaryotes including signal peptidase cleavage sites and gene regulatory sequences. Homologies have been detected between MSPs and immunodominant proteins of Cowdria ruminantium. Several MSPs have been expressed to high level and purified from recombinant Escherichia coli. MSP 1, 2 and 4 have potential for the development of vaccines and MSP3 and 5 for improved diagnostic assays.

Anaplasma↗

Nucleic acid probes as a diagnostic method for tick-borne hemoparasites of veterinary importance.

An increased number of articles on the use of nucleic acid-based hybridization techniques for diagnostic purposes have been recently published. This article reviews nucleic acid-based hybridization as an assay to detect hemoparasite infections of economic relevance in veterinary medicine. By using recombinant DNA techniques, selected clones containing inserts of Anaplasma, Babesia, Cowdria or Theileria genomic DNA sequences have been obtained, and they are now available to be utilized as specific, highly sensitive DNA or RNA probes to detect the presence of the hemoparasite DNA in an infected animal. Either in an isotopic or non-isotopic detection system, probes have allowed scientists to test for--originally in samples collected from experimentally infected animals and later in samples collected in the field--the presence of hemoparasites during the prepatent, patent, convalescent, and chronic periods of the infection in the host. Nucleic acid probes have given researchers the opportunity to carry out genomic analysis of parasite DNA to differentiate hemoparasite species and to identify genetically distinct populations among and within isolates, strains and clonal populations. Prevalence of parasite infection in the tick vector can now be accomplished more specifically with the nucleic acid probes. Lately, with the advent of the polymerase chain reaction technique, small numbers of hemoparasites can be positively identified in the vertebrate host and tick vector. These techniques can be used to assess the veterinary epidemiological situation in a particular geographical region for the planning of control measures.

Anaplasma↗

Molecular basis for vaccine development against anaplasmosis and babesiosis.

Immunization of livestock against the erythroparasitic pathogens Anaplasma marginale, Babesia bigemina, and Babesia bovis with safe and effective killed vaccines is not yet feasible on a practical basis. However, the immune protection afforded by recovery from natural infection and premunition indicates that microbial epitopes capable of inducing immunity exist and that the bovine immune system can be primed appropriately. Induction of protection by immunization with killed parasite fractions, enriched for polypeptides with surface exposed epitopes, supports a focus on surface epitopes, including apical complex organellar epitopes in Babesia, for vaccine development. Cloning, sequencing, and expression of genes encoding these key surface polypeptides has allowed examination of polypeptide function and detailed analysis of epitope conservation in light of genetic polymorphism. In this paper, the characterization of these polypeptides at the epitope level and their roles in inducing protective immunity are reviewed. Definition of these epitopes, in combination with improved understanding of immune mechanisms, provides the basis for development of effective recombinant vaccines against anaplasmosis and babesiosis.

Anaplasma↗

Tick-borne rickettsial diseases: emerging risks in Europe.

Ticks are currently considered the main vectors of human infectious diseases in Europe, particularly since their role in the transmission of the agent of Lyme borreliosis was demonstrated in the 1980s. In the recent years, ticks have also been shown to be the vectors of numerous emerging rickettsial diseases. Although Mediterranean spotted fever (MSF) due to Rickettsia conorii was thought for a long time to be the only tick-borne rickettsial disease prevalent in Europe, five more spotted fever rickettsiae have been described as emerging pathogens in the last decade. Further, cases of infection due to Anaplasma phagocytophilum, the agent of human anaplasmosis (previously known as human granulocytic ehrlichiosis), have been reported throughout Europe. We present here these emerging diseases and discuss other potential threat for the future.

Anaplasma phagocytophilum↗

GIS-facilitated spatial epidemiology of tick-borne diseases in coyotes (Canis latrans) in northern and coastal California.

Ixodes pacificus is the main tick vector for transmission of Anaplasma phagocytophilum and Borrelia burgdorferi to large vertebrates in California. The present study was undertaken in I. pacificus-infested counties in California to examine spatial and temporal relationships among A. phagocytophilum and B. burgdorferi-exposed coyotes with vegetation type and climate. The overall A. phagocytophilum and B. burgdorferi seroprevalences were 39.5% (N=215) and 18.9% (N=148), respectively, with no association with sex. PCR for A. phagocytophilum and B. burgdorferi was negative in all blood and kidney samples. Increased seroprevalence was a positive function of rainfall. Ehrlichial seropositivity was increased in blue-oak foothill pine, montane hardwood, and redwood vegetation regions, and decreased in coastal sagebrush and cropland. Increased exposure to B. burgdorferi occurred in blue oak woodland.

Anaplasma phagocytophilum↗