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PubMed2004

[Participation of Ixodes ricinus developmental stages in transmission of Anaplasma (Ehrlichia) phagocytophila].

The present study constitutes a survey on the prevalence of human granulocytic ehrlichiosis agent, Anaplasma (Ehrlichia) phagocytophila, infecting ticks, Ixodes ricinus. I. ricinus were collected in the spring and autumn of the years 2000-2002 in western Pomerania (Poland), on vegetation using cloth drags. In all, 3340 individuals of I. ricinus were collected from the sampling sites. This total comprised of 511 females, 525 males, and 1998 nymphs. The prevalence of this pathogen in ticks was assessed using the PCR technique (a fragment of 16S rDNA gene amplified with EHR 521 and EHR 747 primers). The overall prevalence of A. phagocytophila in I. ricinus was 3 % in adults but only 1.6% in nymphs.

Anaplasma phagocytophilum↗

PubMed2006

Prevalence of antibodies to serotypes of Bluetongue virus and Anaplasma marginale in Montana feeder cattle: 2002-2003.

In a serologic survey of Montana-source weaned calves and yearling cattle, the apparent prevalence of antibodies to Bluetongue virus was 0.68% and 1.26% in 2002 and 2003, respectively, and to Anaplasma marginale at a positive cutoff at 30% inhibition it was 1.82% and 1.35% in 2002 and 2003, and at a positive cutoff at 42% inhibition it was 0.76% and 0.55% in 2002 and 2003, respectively, suggesting that the risk of importing infected animals was very low.

Anaplasma marginale↗

PubMedDate not supplied

[Seroprevalence of Anaplasma phagocytophilum infection in dogs in Germany].

To determine the prevalence of Anaplasma phagocytophilum in dogs in Germany serum samples from 1124 dogs that were under suspicion of having anaplasmosis were examined. The samples were tested by an indirect immunofluorescence test (OFT) for antibodies to A. phagocytophilum. The geographical origin of positive cases were analysed with an geographic information system. Antibodies to A. phagocytophilum were found in 563 (50.1%) of the tested dogs. 166 dogs came from Saarland, 161 from North Rhine-Westphalia, 134 from Baden-Wuerttemberg, 33 from Bavaria, 22 from Rhineland-Palatinate, 11 from Hamburg, 10 from Brandenburg, 9 from Lower Saxony, 8 from Hesse and Berlin respectively and 1 from Schleswig-Holstein. Clinical signs and laboratory findings of 26 seropositive dogs were analysed. Those dogs showed a low haematocrit, thrombocytopenia and leucocytoses as well as higher values for alkaline phosphatase, ALAT und bilirubin. The clinical signs were lameness in 13 dogs, lethargy in 5, and uveitis in 3 dogs. Rhinitis and lymphadenopathy was found in 2 dogs and retinal detachment with blindness in 1 dog.

Anaplasma phagocytophilum↗

PubMed2006

Natural Anaplasma phagocytophilum infection of ticks and rodents from a forest area of Jilin Province, China.

Polymerase chain reaction integrated with sequence analysis was carried out to investigate the natural Anaplasma phagocytophilum infection in ticks and rodents from a forest area of Jilin Province, China. Four (4.0%) of 100 Ixodes persulcatus and 2 (0.7%) of 286 Dermacentor silvarum ticks collected by flagging vegetation were positive. Nine (8.8%) of 102 rodents were infected, as well as 2 (2.8%) of 71 I. persulcatus parasitizing on 25 rodents. The nucleotide sequences of 1442-bp A. phagocytophilum 16S rRNA gene amplified from rodents and ticks were identical to each other and to that previously reported in Heilongjiang Province of China (GenBank accession no. AF205140), but different from those of other countries. The sequences of 357-bp partial citrate synthase gene from the above specimens were homologic, and varied from known A. phagocytophilum agents. These findings add new information on the ecologic features of A. phagocytophilum and indicate the threat of anaplasmosis in the area.

Anaplasma phagocytophilum↗

PubMed2006

Efficacy of enrofloxacin against severe experimental Anaplasma marginale infections in splenectomized calves.

Four Anaplasma marginale-infected splenectomized calves with greater than 25% parasitized erythrocytes received enrofloxacin at 12.5 mg/kg SC twice, 48 hours apart. Two infected splenectomized calves were designated as untreated controls. A precipitous decline in percent parasitized erythrocytes from 39.13% to less than 1% was observed over 12 days following treatment. However, a self-limiting recrudescence of A. marginale parasites was observed within 30 days after treatment. Untreated control calves became moribund and were euthanized. These data indicate that the regimen of enrofloxacin tested herein ameliorates, but does not eliminate, A. marginale infections in splenectomized calves.

Anaplasma marginale↗

PubMed2006

Comparison of the efficacy of enrofloxacin, imidocarb, and oxytetracycline for clearance of persistent Anaplasma marginale infections in cattle.

This study compared enrofloxacin and imidocarb dipropionate treatments with an oxytetracycline regimen proposed by the World Organization for Animal Health for elimination of persistent Anaplasma marginale infections in cattle. The effect of therapy on competitive ELISA and polymerase chain reaction (PCR) reactivity was also assessed. Twelve A. marginale-infected carrier calves were randomly assigned to groups receiving either enrofloxacin (5 mg/kg IV q24h for 5 days), imidocarb (5 mg/kg IM twice, 7 days apart), or oxytetracycline (22 mg/kg IV q24h for 5 days). One calf infected with an Oklahoma isolate in the imidocarb group and one infected with a Virginia isolate in the oxytetracycline group failed to infect a splenectomized calf following blood subinoculation. Both became competitive ELISA negative by 44 days after treatment, but the imidocarb-treated calf remained PCR positive. None of the tested treatments reliably eliminated persistent A. marginale infections in all cattle. Furthermore, PCR was not a reliable means of determining the success of chemosterilization in calves.

Anaplasma marginale↗

PubMed1990

Intermediate site of development of Anaplasma marginale in feeding adult Dermacentor andersoni ticks that were infected as nymphs.

The development of Anaplasma marginale in midgut epithelial cells was studied in feeding, transmitting adult Dermacentor andersoni ticks. Laboratory-reared ticks experimentally infected as nymphs were allowed to feed from 1 to 9 days on susceptible calves. Gut tissues from ticks were collected on each day they fed (total, 9 days) and were processed for light and transmission electron microscopy. Colonies of A marginale were abundant during the first 6 days of feeding, after which numbers decreased. Colonies were adherent to the basement membrane of gut cells early during feeding, with resultant flattening of the colonies. Colonies also were seen in muscle cells on the hemocoel side of the basement membrane. Morphologic features of A marginale within muscle cells varied and were similar to those observed in gut cells. In addition, however, a large reticulated form in the colonies was observed in muscle cells and appeared to give rise to small particles by budding. Development of A marginale in muscle cells appears to represent an intermediate site of development between those in gut and in salivary glands.

Anaplasma↗

PubMed1979

In vitro cultivation of Anaplasma marginale: growth pattern and morphologic appearance.

Anaplasma marginale propagated in vitro showed an increasing rate of replication in a sequence of three experiments. The changes in the percentage of parasitized erythrocytes and identification of the organism in culture were monitored by the Giemsa-staining and the direct fluorescent antibody techniques. The ultrastructure of the organism in culture also was determined. The percentage of parasitized erythrocytes increased more than three times in the first experiment during a period of 8 days, and about ten times in the second experiment during a period of 14 days. After 8 days of observation of the primary culture in the third experiment, the trend of growth was more rapid than in the first and second experiment. The viability of the organism was verified by inoculation of susceptible calves, using 13- and 33-day cultures of the first and the second experiments, respectively. The primary cultures were subcultured twice by dilution with normal bovine erythrocytes.

Anaplasma↗

PubMed1975

Exit of Anaplasma marginale from bovine red blood cells.

Incubation of Anaplasma marginale-infected bovine red blood cells in a flow culture system resulted in a decrease of observable marginal bodies. The decrease was twice the degree of hemolysis, indicating that marginal bodies may leave red blood cells without concomitant lysis of the host cell.

Anaplasma↗

PubMed1988

Preliminary studies of the development of Anaplasma marginale in salivary glands of adult, feeding Dermacentor andersoni ticks.

On each day of feeding on susceptible calves, salivary glands obtained from groups of adult ticks that transmitted Anaplasma marginale were examined for A marginale colonies by use of light microscopy and transmission electron microscopy. On day 8 of feeding, salivary glands were examined, using fluorescein-labeled antibody and methyl green-pyronine stain. Use of fluorescein-labeled antibody consistently revealed small numbers of fluorescent foci in salivary gland acinar cells obtained from ticks that had fed for 8 days. Colonies of A marginale were seen by transmission electron microscopy only in salivary gland acini of male ticks; these colonies could not be identified, using light microscopy, in companion 1-micron plastic sections stained with Mallory stain. Methyl green-pyronine stain, used commonly to detect theilerial parasites in tick salivary glands, did not differentiate A marginale from cytoplasmic inclusions normally found in salivary gland acinar cells.

Anaplasma↗

PubMed1989

Anaplasma marginale in tick cell culture.

Anaplasma marginale was propagated in a tick cell line derived from Dermacentor variabilis embryos. The rickettsial organism was identified and monitored in culture by transmission electron microscopy and the indirect immunofluorescence technique, using specific monoclonal antibodies. Inoculation of the embryonic tick cell line with midguts of infected adult ticks (culture 1), nymphal ticks (culture 2) and adult ticks that were infected as nymphs and dissected as adults (culture 3) resulted in 3 continuous cultures of A marginale. Culture 1 had been maintained through 22 passages over a 11-month period; cultures 2 and 3 had been maintained for 18 passages over a 9-month period. Growth of A marginale in the cell line began in the area of the nuclear membrane at approximately 4 days after inoculation or transfer. Thereafter, the organisms were observed in inclusions scattered throughout the cytoplasm of the host cells. Maximal growth of the organism occurred at 7 to 14 days, after which numbers of inclusions rapidly decreased to minimal or undetectable levels. The organism began new cycles of growth with each 1:5 to 1:10 split and transfer of the host cells. Electron microscopy of recently infected cells revealed a morphology of the organism that closely resembled that observed in marginal bodies of infected erythrocytes. After several passages, A marginale organisms had a varied morphology and resembled the organism described in midgut cells of naturally infected ticks.(ABSTRACT TRUNCATED AT 250 WORDS)

Anaplasma↗

PubMed1989

Transstadial and attempted transovarial transmission of Anaplasma marginale by Dermacentor variabilis.

Transstadial and transovarial transmission of Anaplasma marginale by Dermacentor variabilis were attempted with with ticks exposed to the organism once by feeding as larvae or nymphs, and twice by feeding as larvae and nymphs. Typical colonies of A marginale were in gut tissues of adults that were infected as larvae, larvae and nymphs, and as nymphs; repeated exposure of ticks did not appear to result in an increase in the number of colonies in the gut of subsequently molted adults nor did it affect severity of the clinical disease that developed in cattle they fed on. In contrast, colonies of A marginale were not found in the midgut epithelium of unfed nymphs exposed as larvae, even though companion nymphs transmitted the parasite, causing severe clinical anaplasmosis in susceptible calves. The organism was not transmitted transovarially by F1 larvae or nymphs from the groups exposed as parent larvae, nymphs, larvae and nymphs, and as adults. Some of the calves fed on by F1 progeny had a few erythrocytic marginale bodies that looked suspiciously like A marginale, as well as postchallenge exposure prepatent periods that were longer than other calves in the transovarial transmission study. Sera from these calves were tested for antibody to A marginale, using a highly sensitive immunoblot technique. Antibodies were not detected in any of the sera.

Anaplasma↗

PubMed1989

Detection of colonies of Anaplasma marginale in salivary glands of three Dermacentor spp infected as nymphs or adults.

Salivary glands from males of 3 Dermacentor species (D andersoni, D variabilis and D occidentalis) that were infected with either the Virginia or Idaho isolate of Anaplasma marginale as nymphs or adults were examined for colonies of A marginale by use of light and electron microscopy. Prior to dissection of salivary glands, exposed ticks were held at 25 C for 15 to 18 days, followed by a 3-day incubation at 37 C. Ticks of 2 species transmitted A marginale to calves; the third tick species was confirmed infected by demonstration of typical colonies in tick gut cells, but transmission was not attempted; Colonies of A marginale were seen with light microscopy in salivary glands of all 3 species of ticks; they were located in acinar cells that contained simple granules. Colonies varied morphologically from small, compact ones to larger structures that contained distinct organisms and often were adjacent to the host cell nucleus. Electron microscopy confirmed that the colonies were rickettsial organisms. Morphologic features of A marginale varied and included reticulated forms, forms with electron-dense centers, and small particles; these various forms were similar to those described previously in midgut epithelial cells of ticks. We believe that the organism seen within tick salivary glands may replicate in the glands before its transmission to the vertebrate host.

Anaplasma↗

PubMed1987

Tick transmission of Anaplasma centrale.

Anaplasma centrale was isolated from a field collection of Rhipicephalus simus. Transstadial transmission of A. centrale with adult ticks was demonstrated, but the infection was not carried transovarially. Ticks from this collection were subsequently reared as a non-infected, laboratory strain. It was proved that the Onderstepoort live blood vaccine strain of A. centrale, isolated by Theiler in 1911, is still tick transmissible after more than 75 years of needle passage through cattle in the laboratory. Attempts to demonstrate transstadial transmission of the vaccine strain with Boophilus decoloratus and Boophilus microplus failed.

Anaplasma↗

PubMed1987

Persistence of colonies of Anaplasma marginale in overwintering Dermacentor variabilis.

Dermacentor variabilis were infected as nymphs with Anaplasma marginale by allowing the ticks to feed on a single infected donor calf. Two weeks after molting to the adult stage, the ticks were allotted into 1 of 3 groups and were allowed to overwinter at room temperature (25 C) in the laboratory (group 1), cold storage (4.5 C) in the laboratory (group 2), or outdoors in leaf litter (group 3). Persistence of A marginale was assessed by determining density of colonies (number of colonies/0.1 mm2 of gut tissue examined) in tick gut specimens at 3, 5, 7, 9, and 12 months after molting to the adult stage. Colonies of A marginale were found in all groups at every density evaluation period. Highest colony densities were observed uniformly in specimens collected at month 7 (May); densities decreased at month 9 and were lowest at month 12. Statistical analysis indicated that ticks subjected to cold storage and to outdoor conditions had similar colony densities of A marginale; the density curve in these 2 groups indicated significant quadratic effects over time, with peak densities in May. Mean colony density in ticks kept at room temperature fit a different quadratic equation. The morphologic data indicated that A marginale overwinters in Dermacentor variabilis, and that increasing numbers of organisms are found from January to May.

Anaplasma↗

PubMed1986

Longevity of colonies of Anaplasma marginale in midgut epithelial cells of Dermacentor andersoni.

Colonies of Anaplasma marginale in midgut epithelial cells of experimentally infected Dermacentor andersoni were studied in adult ticks 1, 3, and 6 months old. Longevity of the parasite in ticks was assessed by evaluating its infectivity for splenectomized calves; calves were exposed by feeding ticks and by inoculation of tick gut homogenates. Longevity was also evaluated by determining size, type, and density of colonies in male and female ticks. The effect of incubation (2.5 days at 37 C) on colony density was also examined for ticks at each age period. All methods used to assess longevity of A marginale in ticks (tick transmission, calf inoculation, and histologic studies) indicated a decrease of the numbers of organisms in 6-month-old ticks. Furthermore, when tick gut homogenates from 6-month-old nonincubated ticks were not infectious for susceptible calves, incubation of ticks before dissection restored infectivity of homogenates. Colonies of A marginale were detected in gut tissues of 6-month-old ticks that were not infective; therefore, infectivity of ticks could not be confirmed merely by presence of A marginale colonies.

Anaplasma↗

PubMed1986

Percutaneous infection of nymphal Dermacentor andersoni with Anaplasma marginale.

Newly replete nymphal Dermacentor andersoni (principals) were percutaneously exposed to Anaplasma marginale by injection of either intact or lysed infected bovine erythrocytes. Control nymphs were fed on calves with anaplasmosis. The subsequently molted adults were examined for infection by light microscopy, and companion ticks were tested for infectivity by allowing them to feed on susceptible calves. When they fed as adults, both control ticks and percutaneously inoculated principals transmitted A marginale to susceptible calves. Prepatent periods in calves varied according to the method by which nymphs were infected. Colonies of A marginale were found in all ticks that acquired infection by feeding, but colonies were not observed in any ticks exposed percutaneously. The possible developmental cycle of A marginale in artificially infected ticks is discussed.

Anaplasma↗

PubMed1986

Transmission of Anaplasma marginale Theiler by males of Dermacentor andersoni Stiles fed on an Idaho field-infected, chronic carrier cow.

The role of ticks and carrier cattle in epizootics of bovine anaplasmosis was further clarified by demonstrating unequivocally, for the first time, that male ticks fed on a chronic carrier cow naturally infected with Anaplasma marginale can transmit this parasite intrastadially and biologically when subsequently fed on susceptible cattle. These data indicate that field epizootics of acute anaplasmosis may be initiated by males of tick vector species that feed on carrier cattle and subsequently transfer to susceptible cattle.

Anaplasma↗