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Intrastadial and interstadial transmission of Anaplasma marginale by Boophilus annulatus ticks in cattle.

The 1-host tick Boophilus annulatus was found to transmit anaplasmosis in cattle transstadially. Anaplasma marginale was invariably transmitted when ticks that had been pulled off Anaplasma-infected calves either after 7 days (as fully engorged larvae) or after 14 to 15 days (as fully engorged nymphs) were transferred within 4 days to susceptible calves. Three morphologically different A marginale isolates, 1 round (tailless) and 2 with different types of appendages (tailed) were transmitted by the ticks. These findings might explain the overlap of the geographic distribution of the disease and that of Boophilus spp in some areas of the world.

Anaplasma↗

Observations on the use of Anaplasma centrale for immunization of cattle against anaplasmosis in Zimbabwe.

A total of 93 Bos taurus cattle was used in pen trials to compare vaccine stocks of Anaplasma centrale from South Africa and Australia (which stock came from South Africa in 1934) in protecting against three virulent field isolates from clinical Anaplasma marginale infections. In addition, field observations were made on the use of a vaccine, prepared from the Australian stock, in over 9553 cattle of mixed age and breeds on 16 co-operator farms and at one communal dip. The results of the pen trials indicated that the two vaccine stocks were comparable and that neither provided adequate protection against two of the three isolates of A. marginale. The field observations indicated that the vaccine was highly infective and produced mild reactions in most recipient cattle, and that users were generally satisfied with the vaccine. These somewhat conflicting results are discussed in the context of observations in Australia and future vaccination against anaplasmosis in Zimbabwe.

Analysis of Variance↗

Testicular degeneration and libido loss in beef bulls experimentally inoculated with Anaplasma marginale.

Four sexually mature virgin beef bulls were inoculated with whole blood from a known Anaplasma marginale carrier. All four bulls became infected and experienced some degree of anemia and testicular degeneration, while uninoculated controls remained normal. Testicular degeneration was confirmed by electron microscopy, histopathology, and semen evaluation. Further observations established that the inoculated bulls exhibited a loss of libido, while controls reacted in a normal manner to a cow in estrus. Anaplasma marginale was not present in semen of inoculated bulls.

Journal Article↗

Conserved recombinant antigens of Anaplasma marginale and Babesia equi for serologic diagnosis.

The competitive inhibition ELISA (CI-ELISA) format overcomes problems associated with antigen purity since the specificity of the CI-ELISA depends solely on the monoclonal antibody (mAb) used. Therefore, the CI-ELISA format is well suited for use with recombinant antigens. Molecular clones expressing a conserved 19 kDa protein of Anaplasma marginale and a 34 kDa protein of Babesia equi were derived and characterized. The 19 kDa A. marginale protein, conserved in all recognized Anaplasma species, and present in the infected tick salivary gland, was reactive with all bovine immune sera tested. The 34 kDa B. equi protein contains a protein epitope bound by antibody in equine immune sera from 19 countries. Monoclonal antibodies reactive with these proteins were derived and applied with recombinant copies of the 19 kDa A. marginale and 34 kDa B. equi proteins in a CI-ELISA format.

Amino Acid Sequence↗

Surveillance of Egyptian fleas for agents of public health significance: Anaplasma, Bartonella, Coxiella, Ehrlichia, Rickettsia, and Yersinia pestis.

Serologic surveys in Egypt have documented human and animal exposure to vector-borne bacterial pathogens, but the presence and distribution of these agents in arthropods has not been determined. Between July 2002 and July 2003, fleas were collected from 221 mammals trapped in 17 cities throughout Egypt. A total of 987 fleas were collected, representing four species (Ctenocephalides felis, Echidnophaga gallinacea, Leptopsylla segnis, and Xenopsylla cheopis); 899 of these fleas were X. cheopis from rats (Rattus spp.). Fleas were tested for DNA from Anaplasma spp., Bartonella spp., Coxiella burnetii, Ehrlichia spp., Rickettsia spp., and Yersinia pestis. Rickettsia typhi, the agent of murine typhus, was detected in X. cheopis and L. segnis from rats from nine cities. A spotted-fever group Rickettsia sp. similar to "RF2125" was detected in E. gallinacea, and two unidentified spotted fever group Rickettsia were detected in two X. cheopis. Novel Bartonella genotypes were detected in X. cheopis and L. segnis from three cities. Coxiella burnetii was detected in two fleas. Anaplasma, Ehrlichia, and Y. pestis were not detected.

Animals↗

Evaluation of jackrabbits as nonruminant hosts for Anaplasma marginale.

Two black-tailed jackrabbits (Lepus californicus), 1 splenectomized and 1 intact, were inoculated with 0.2 ml of a 1:5 dilution of a Florida Anaplasma marginale stabilate. Five months later, both hares were inoculated with 1 ml of whole blood from a calf with acute anaplasmosis. Neither hare developed any signs of clinical anaplasmosis. Pooled blood (7 ml) from these jackrabbits which was inoculated into 2 Anaplasma-susceptible, splenectomized calves failed to induce hematologic or serologic signs of anaplasmosis for at least 90 days. Two susceptible, splenectomized calves were inoculated with 35 ml of pooled whole blood from 9 wild-collected black-tailed jackrabbits from a known anaplasmosis enzootic area. Both steers remained free of anaplasmosis signs for 90 days.

Anaplasmosis↗

Ultrastructural aspects of intraerythrocytic development of a Venezuelan strain of Anaplasma marginale.

Blood samples from splenectomized calves infected with a Venezuelan strain of Anaplasma marginale were studied by electron microscope. A. marginale appears to invade erythrocytes by the invagination of the host cell plasma membrane. The parasite reproduces by binary or multiple fission, changing from a rounded to an irregularly polyhedral form during and after division. The mature Anaplasma, within the host red cell modifies the erythrocytes cytoplasma and external membrane producing pores through which the parasite can escape without lysing the host erythrocyte.

Anaplasmataceae↗

Anaplasma ovis in Idaho sheep.

Blood samples from 16 anaplasmosis complement-fixation (CF) positive sheep from a flock near Cambridge, Idaho, were inoculated into 3 intact sheep and 3 splenectomized calves. The 3 sheep became CF positive, and 2 developed parasitemia. None of the 3 calves had signs of anaplasmosis, but all became ill when they were subsequently challenge exposed with blood from known carriers of Anaplasma marginale. After a 75-day observation period, blood from the recipient intact sheep was subinoculated into 3 splenectomized sheep. These splenectomized sheep developed clinical anaplasmosis and became CF positive. In only 1 sheep was there a positive rapid card agglutination test. Therefore, the rapid card agglutination test was not considered to be diagnostic for anaplasmosis carrier status in sheep. Electron microscopic studies were performed on infected sheep erythrocytes to determine the morphologic structure of the organism involved. The causative organism was determined to be Anaplasma ovis.

Anaplasmosis↗

Influence of dexamethasone on the recrudescence of Anaplasma marginale in splenectomized calves.

Dexamethasone was administered at the dose rate of 0.2 mg/kg of body weight to 11 splenectomized Anaplasma-carrier calves (groups 1 and 3) on Monday, Wednesday, and Friday for 3 weeks. Observations were made on these calves and on 7 nontreated, comparable calves (group 2) to determine the influence of treatment on carrier infections. Dexamethasone treatment was associated in every instance with an exacerbation of the Anaplasma parasitemia and a decrease in packed red cell volume. The episode of acute anaplasmosis was of short duration, resembling the primary response, except that complement-fixation response did not increase accordingly. Serum protein electrophoresis of serums from 4 calves (group 3) undergoing the drug-induced response failed to show any significant change during the 3-week treatment period, but did show a significant increase in gamma-globulin immediately after treatment.

Anaplasmosis↗

Tick-borne diseases of sheep and goats caused by Babesia, Theileria or Anaplasma spp.

A review is given on the Babesia, Theileria, and Anaplasma species infecting sheep and goats. B. ovis is the most important disease agent. It is transmitted by Rhipicephalus bursa, R. turanicus, Hyalomma anatolicum excavatum, and probably by R. evertsi evertsi B. ovis is widely spread in southern Europe, the Middle East, and central Asia. Its geographical distribution in South and East Asia and in Africa is widely unknown. B. motasi obviously represents several nosodemes in separate regions. It is not pathogenic for intact sheep in northern Europe, whereas it is probably more pathogenic than B. ovis in India and northern Africa. The known vectors of B. motasi are Haemaphysalis punctata and R. bursa. Theileria hirci is transmitted by H. a. anatolicum but occurs outside the distribution area of this tick. Malignant theileriosis of sheep and goats is an important disease in Iraq, Iran, and India. An attenuated macroschizont vaccine is successfully being used in Iran. Anaplasma ovis is transmitted by R. bursa and probably other ticks in the Old World and by Dermacentor andersoni in the New World. A. ovis is widely spread in the Old World. Outbreaks occur only under extreme conditions. The identity of the tick-borne disease agents of sheep and goats and of their vector ticks is uncertain in many regions of the Old and the New World.

Anaplasmosis↗

Detection of Babesia and Anaplasma species in rabbits from Texas and Georgia, USA.

Rabbits have been shown to harbor a suite of zoonotic organisms, including a Babesia species, Borrelia burgdorferi, and Anaplasma phagocytophilum. In this study, we conducted a molecular survey for various tick-borne pathogens in three species of rabbits from Texas and Georgia. Of 18 black-tailed jackrabbits (Lepus californicus) tested from Texas, six (28%) were polymerase chain reaction (PCR) positive for Babesia, and nucleotide sequencing revealed two distinct species or strains. Two jackrabbits were infected with a Babesia species or strain (Babesia sp. A) that was nearly identical (99.9%) to a piroplasm previously detected in humans from Washington state, and the remaining four jackrabbits were infected with a Babesia species (Babesia sp. B) that was most similar (99.7%) to a Babesia species detected in cottontail rabbits from Massachusetts and humans from Kentucky and Missouri. Eleven (61%) black-tailed jackrabbits were positive for A. bovis, and one was positive for A. phagocytophilum. Two of four desert cottontails (Sylvilagus audubonii) from Texas were positive for the Babesia sp. B, and one desert cottontail each was positive for A. bovis and A. phagocytophilum. One of these desert cottontails was coinfected with the Babesia sp. B and A. phagocytophilum, and five jackrabbits were coinfected with Babesia species and A. bovis. Of 19 eastern cottontails (S. floridanus) from Georgia, only one (5.3%) was positive for A. phagocytophilum, and three (15.8%) were positive for A. bovis. No rabbits from Texas or Georgia were positive for Borrelia species. The only tick species detected on the Texas and Georgia rabbits was the rabbit tick, Haemaphysalis leporispalustris. These data extend the geographic and host range of these pathogens, and because both the Babesia species and A. phagocytophilum are potential zoonotic pathogens, it is important to be aware that these organisms are enzootic in parts of the southern United States.

Anaplasma↗

Equine and canine Anaplasma phagocytophilum strains isolated on the island of Sardinia (Italy) are phylogenetically related to pathogenic strains from the United States.

The presence of Anaplasma phagocytophilum, a tick-transmitted zoonotic pathogen, was investigated in Sardinia using a molecular approach. Phylogenetic analysis revealed that Sardinian strains are genetically distinct from the two lineages previously described in Europe and are closely related to strains isolated in different areas of the United States.

Anaplasma↗

PCR detection of granulocytic Anaplasma and Babesia in Ixodes ricinus ticks and birds in west-central Poland.

The aim of the study was to establish the role of forest birds as reservoirs of Anaplasma phagocytophilum and Babesia spp. in Wielkopolski National Park. A total of 108 birds from 9 species were collected between May-September 2002. Blood samples were taken from 84 specimens and 442 individuals of the common tick, Ixodes ricinus, were collected from the birds. The 73 additional ticks were collected from vegetation. PCR amplification of a fragment of the epank 1 gene and 18S rRNA gene was used for detection of A. phagocytophilum and Babesia spp. DNA, respectively. Pathogen DNA was not detected in any of the blood samples or ticks collected from birds. On the other hand, 3 ticks collected from vegetation (4.1% of all examined specimens) were positive for A. phagocytophilum DNA. In spite of the high level of infestation of birds by I. ricinus, it is clear that they do not constitute a competent reservoir of A. phagocytophilum and Babesia in WNP. Additionally, I. ricinus is not a significant vector in this area.

Anaplasma↗

Frontline: control of Anaplasma phagocytophilum, an obligate intracellular pathogen, in the absence of inducible nitric oxide synthase, phagocyte NADPH oxidase, tumor necrosis factor, Toll-like receptor (TLR)2 and TLR4, or the TLR adaptor molecule MyD88.

Anaplasma phagocytophilum is an obligate intracellular bacterium that is related to rickettsial organisms and replicates in the hostile environment of neutrophils. Previous studies with SCID mice suggested that T and/or B cells are required for its control in vivo. Here, we used mice deficient for Toll-like receptor (TLR)2 and TLR4, MyD88, tumor necrosis factor, inducible nitric oxide synthase, or phagocyte NADPH oxidase (gp91(phox-/-)) to define the pathways that are critical for the recognition and the killing of this pathogen. Whereas SCID mice developed a 60-fold higher bacterial load in the blood compared to wild-type mice and succumbed to infection, all other gene-deficient mouse strains were fully capable in overcoming a systemic infection with A. phagocytophilum. From these data we conclude that effector mechanisms that are crucial to the defense against numerous other intracellular pathogens are dispensable for the control of A. phagocytophilum.

Adaptor Proteins, Signal Transducing↗

Preliminary studies on the effect of Anaplasma marginale antibodies ingested by Dermacentor andersoni ticks (Acari:Ixodidae) with their blood meal on infections in salivary glands.

The effect of Anaplasma marginale antibodies ingested with the tick blood meal was tested on infected male ticks that were allowed to feed on cattle immunized with the erythrocytic stage of A. marginale. The experiments were done in two trials. Trial 1 was done using splenectomized calves (two calves per treated and control groups) while ticks in trial 2 were fed on intact yearling cattle (four cattle per treated and control groups). The cattle were immunized with purified outer membrane proteins of erythrocyte-derived A. marginale using saponin (trial 1) or monophosphoryl lipid-A-trehalose dicorynomycolate adjuvant (trial 2). The corresponding control cattle received adjuvant only. All cattle were challenged using Dermacentor andersoni males infected as adults that were allowed to feed for 7 days. In trial 1, the ticks were allowed to feed a second time on susceptible calves to test whether exposure of ticks to immunized cattle affected their ability to transmit anaplasmosis. Infections in fed ticks were monitored by determining the infection rates in salivary glands with an A. marginale-specific RNA probe and light microscopy. Vaccine-derived antibodies ingested with the tick blood meal did not appear to affect the development of A. marginale in previously infected ticks. The infection rates in the salivary glands were not significantly different among ticks fed on immunized versus adjuvant control cattle. When the vaccine-exposed ticks in trial 1 were allowed to feed a second time on susceptible calves, the resulting clinical symptoms of anaplasmosis were similar to those of the controls. There was no statistically significant effect of tick exposure to the anti-erythrocytic stage antibody on the development of salivary gland infection or transmission of A. marginale by ticks.

Anaplasma↗

Anaplasma marginale: failure of sera from immune cattle to confer protection in passive-transfer experiments.

High levels of immunity to Anaplasma marginale were induced in cattle either by vaccination using sonically disrupted A. marginale-infected erythrocytes or by repeated infection with different strains of the rickettsia. In both instances, high levels of anti-A. marginale antibody were detected in the sera of the immune cattle by immunoblotting. Serum from one animal that had been made immune by repeated infection was transferred intravenously to A. marginale-susceptible calves (three non-splenectomised and two splenectomised) undergoing initial A. marginale infection at serum doses of 2-10 ml/kg. Neither the course nor the outcome of infection as indicated by the parasite levels attained or the level of anaemia induced was altered in the calves that received the immune serum relative to the course or outcome of infection in control calves (two non-splenectomised and two splenectomised) that received serum from an two splenectomised) that received serum from an A. marginale-naive donor animal. In a similar experiment, a pool of sera from four steers that had been vaccinated with sonically disrupted A. marginale initial bodies was transfused into two intact A. marginale-susceptible calves during the early stage of A. marginale infection at a dose of 10 ml/kg. No difference was observed in the course or outcome of infection in these calves relative to the course or outcome of infection in the two non-splenectomised calves that were transfused with non-immune serum.(ABSTRACT TRUNCATED AT 250 WORDS)

Anaplasma↗

Enzyme-linked immunosorbent assay using solubilised antigen for detection of antibodies to Anaplasma marginale.

An enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies against Anaplasma marginale. A marginale bodies were separated from parasitised erythrocytes by a modified nitrogen decompression method, sonicated, then solubilised with Triton X-100 and used as the ELISA antigen. In this ELISA system the required amount of antigen protein was 16.2 ng for each well. In the course of experimental infections, of calves, significant antibody levels were detected by ELISA and the complement fixation test (CFT) at almost the same time. Antibodies against A. marginale were detectable for longer periods using the ELISA than using the CFT. Sera from calves infected with Babesia bigemina, B. bovis, B. ovata, Theileria sergenti and Eperythrozoon wenyoni gave no reaction; however, antisera against A. centrale did react with the A. marginale ELISA antigen.

Anaplasma↗

Characteristics of an attenuated Anaplasma marginale of deer origin as an anaplasmosis vaccine.

A fully virulent field Anaplasma marginale isolate, passaged to a splenectomised white tailed deer, lost virulence for cattle after 18 months in the non-bovine host. This attenuated A. marginale produced a mild response in four 18-month-old Holstein steers characterised by an average 2.4% parasitaemia, a 13% drop in PCV and a positive serological response. All vaccinated steers were immune to challenge with a fully virulent, heterologous A. marginale when compared with non-vaccinated controls.

Anaplasma↗