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Prevalence of blood parasites in cattle from wilayates of Annaba and El Tarf east Algeria.

Between June and September 2002, a preliminary study was conducted to assess the prevalence of blood parasites of cattle in eastern Algeria. Fifty-four bovines of different genotypes were submitted to clinical examination. From each animal, blood smears were made and stained by Giemsa. Four species of parasites, namely Theileria annulata, T. orientalis, Babesia bovis and Anaplasma marginale were encountered. Fifty animals carried single or multiple infections with blood parasites and four were found negative. The rate of single infections (72.3%, n = 39) was almost three times higher than multiple infections (20.3%, n = 11). The high percentage of single infections was recorded with T. annulata (53.7%). However single infection with Anaplasma marginale (7.4 %), B. bovis (5.6%) and T orientalis (5.6%) were very low compared to T. annulata infection. The rates of mixed infection were as follows: T. annulata/A. marginale 9.3%, T. annulata/T. orientalis 5.6%, A. marginale/T. orientalis 3.7% and B. bovis/A. marginale 1.9%.

Algeria↗

[First report of Anaplasma bovis (Donatien and Lestoquard, 1936) Dumler et al. (2001) at micro region of Campos dos Goytacazes, State of Rio de Janeiro, Brazil].

The first report of Anaplasma bovis was pointed out at the micro region of Campos dos Goytacazes, State of Rio de Janeiro, Brazil. Blood smears were done from peripherical blood vessels from 151 calves. Five calves (3.31%) were positive for A. bovis. Symptoms were characterized by debility, fever, pale mucous membranes, and enlargement of prescapular lymph nodes.

Anaplasmosis↗

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↗

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↗

Concurrent infection with Theileria buffeli caused depression of parasitaemia in Babesia bovis and Anaplasma centrale infections in splenectomised calves but not in B bigemina infections.

In five experiments undertaken in splenectomised calves it was found that Theileria buffeli infections depressed the parasitaemias of superimposed Babesia bovis and Anaplasma centrale infections, but not B bigemina infections. The course of A centrale infections did not appear to be affected by a concurrent Eperythrozoon teganodes infection.

Anaplasmosis↗

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↗

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↗

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↗

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↗

Infectivity and antigenicity of Anaplasma marginale from tick cell culture.

The infectivity and immunogenicity of Anaplasma marginale grown in a tick cell culture from embryonic Dermacentor variabilis ticks were assessed in splenectomized and intact calves, respectively. Culture 1 consisted of the cell line inoculated with midguts of adult ticks infected with the Mississippi isolate of A marginale and dissected 5 to 10 days after repletion and detachment from an experimentally infected calf. Cultures 2 and 3 consisted of the cell line inoculated with midguts of ticks infected with the Virginia isolate of the organism. Inoculum for culture 2 was derived from nymphal ticks dissected 5 to 10 days after repletion and detachment from the infected calf; inoculum for culture 3 was midguts from adult ticks that were fed as nymphs, allowed to molt in the laboratory and dissected 21 to 24 days after molting. In trial 1, cultures 1, 2, and 3 were maintained at pH 6.9 and incubated at 28 C; in trial 2, cultures 1 and 3 were maintained at pH 7.4 and incubated at either 28 C or 37 C. Cultures 1, 2, and 3 failed to induce infection when injected IV and SC into 6 calves in 2 separate trials. Pre-challenge sera from these calves reacted with 2 purified Anaplasma antigens in the ELISA, but failed to react in the complement-fixation test. Results of a trial to use cultures 1 and 3 in combination with an oil-in-water adjuvant to immunize intact calves against A marginale were inconclusive. However, pre-challenge sera from immunized calves reacted with the 2 purified Anaplasma initial body antigens in the ELISA but failed to react in the complement-fixation text.(ABSTRACT TRUNCATED AT 250 WORDS)

Anaplasma↗

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↗

Fitting mixture models to grouped and truncated data via the EM algorithm.

The fitting of finite mixture models via the EM algorithm is considered for data which are available only in grouped form and which may also be truncated. A practical example is presented where a mixture of two doubly truncated log-normal distributions is adopted to model the distribution of the volume of red blood cells in cows during recovery from anemia.

Algorithms↗

The persistence of colostral Anaplasma antibodies and incidence of in utero transmission of Anaplasma infections in calves under laboratory conditions.

Twenty-six calves, born from 25 Anaplasma-infected, intact and splenectomized cows, from a herd kept under strict tick-free laboratory conditions, were monitored for the presence of Anaplasma antibodies, using the rapid card agglutination test. Serum was collected at birth, weekly for 12 weeks, and then monthly for approximately 6 months. Specific antibodies passively acquired could be detected in calf sera for an average period of 8 weeks after birth. Calves that remained positive for longer than 12 weeks were suspected of having contracted in utero infections. Infection of the calves was confirmed by splenectomy. It was concluded that 4 calves in Group I contracted in utero infections. Two of the dams were chronically infected, whilst the other 2 underwent acute primary reactions during the 1st and 2nd trimesters of gestation, respectively. Subsequently all calves born from infected cows in this tick-free herd were serologically screened before being splenectomized at an average age of 8 months. Out of 50 cows, 8 in utero infected calves were identified serologically and this finding was confirmed through splenectomy or subinoculation of blood. Both Anaplasma centrale and Anaplasma marginale were carried transplacentally. Splenectomized and intact cows, chronically infected or undergoing primary reactions during the 1st, 2nd or 3rd trimester of gestation, produced infected calves. A 15,6% incidence of in utero transmitted infections were observed amongst 77 calves under these conditions. None of the 13 splenectomized cows, undergoing primary A. centrale infections during gestation, aborted. Clinical signs of disease were not observed in any of the 12 in utero infected calves prior to splenectomy. The implications of these findings are discussed.

Anaplasmosis↗

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↗

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↗