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Biomedical subjects

S A Ewing

Publications and source records attributed to S A Ewing.

At least 91 records · Page 5Linked to original sources

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↗

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↗

Transmission of Anaplasma marginale by adult Dermacentor andersoni during feeding on calves.

Laboratory-reared Dermacentor andersoni ticks experimentally infected as nymphs with Anaplasma marginale were allowed to feed as adults from 1 to 9 days on susceptible, splenectomized calves to determine when, during feeding, the hematozoan was transmitted from ticks to cattle. In experiment 1, ticks were allowed to feed on calves for 1, 2, 3, 4, 5, or 6 days and anaplasmosis did not result. The same calves were used for experiment 2, and ticks were allowed to feed for 1, 3, 6, 7, 8, or 9 days and anaplasmosis occurred in all calves on which ticks fed for greater than or equal to 6 days. In 2 trials in experiment 3, ticks were allowed to feed on calves for 1 to 9 days. Anaplasmosis developed only in calves on which ticks fed for 7, 8, or 9 days. The prepatent periods shortened with longer tick feeding, and linear regression analysis of combined prepatent periods of both trials of experiment 3 indicated a significant (P = 0.05) slope with an estimated daily decrease of 7.75 days from day 7 to 9 of feeding. There was no apparent correlation between length of tick feeding and severity of clinical signs in those calves that developed anaplasmosis. Seemingly, A marginale can be transmitted to cattle by adult D andersoni ticks no earlier than the 6th or 7th day of feeding.

Anaplasmosis↗

Demonstration of the inclusion appendage of Anaplasma marginale in nymphal Dermacentor andersoni.

Dermacentor andersoni nymphs were placed in stockinettes and allowed to feed on a splenectomized calf with experimentally induced anaplasmosis when the parasitemia was 3%-5%. Nymphs were selected on each of the 6 days of feeding and every 5 days from repletion through molting to the adult stage (25 days postrepletion); they were killed and midgut tissues were processed and examined by light and electron microscopies. No stages of A marginale were seen in tissues of feeding ticks. Visualization of individual components of gut contents was difficult owing to presence of the concentrated, electrondense blood meal containing hemoglobin. Inclusion appendages were observed in midgut tissues of nymphs at 5 and 10 days postrepletion, but not at 20 or 25 days. The morphology of the appendages was similar to that described for inclusion appendages commonly associated with anaplasmal inclusions in bovine erythrocytes. Some appendages were free in the lumen of the midgut and occurred either alone or with clusters of small vesicular particles. Occasionally, initial bodies like those generally found in bovine erythrocytes were seen with the appendage, but most of them were swollen and appeared to be degenerating. Frequently, inclusion appendages were observed attached to the luminal surface of the midgut cell membrane by a blunt, electron-dense attachment complex. The attachment of the appendage appeared to be extracellular, with the pointed end extending into the lumen. Often, small particles were observed immediately across the cell membrane from where the appendages were attached; the small particles appeared to be generated from the appendage itself and to have passed through the membrane of the midgut cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Anaplasma↗

Demonstration of Anaplasma marginale in hemolymph of Dermacentor andersoni by animal inoculation and by fluorescent-antibody technique.

Hemolymph was collected from adult Dermacentor andersoni Stiles that had been infected with Anaplasma marginale Theiler as nymphs. Before hemolymph was collected, the adult ticks were either incubated and unfed at 37 C for 2.5 days or fed for 6 days on sheep. Hemolymph collected from groups of 100 ticks was inoculated into susceptible splenectomized calves. Smears of hemolymph from the same groups of ticks were prepared for examination by fluorescent antibody technique. Hemolymph from incubated ticks caused anaplasmosis in 2 of 4 trials, and hemolymph from feeding ticks caused anaplasmosis in 4 of 4 trials. Moderately fluorescing bodies were demonstrated in some hemocytes from incubated ticks, whereas hemocytes from feeding ticks contained numerous clusters of brightly fluorescing bodies. Fluorescing bodies were not observed in hemocytes from control ticks.

Anaplasma↗

Demonstration of colonies of Anaplasma marginale in the midgut of Rhipicephalus simus.

Rhipicephalus simus nymphs were allowed to feed on a cow experimentally infected with the BW-strain of Anaplasma marginale from Republic of South Africa, and they were studied as adults. Colonies were demonstrated by light microscopy in midgut epithelial cells of adult ticks that were unfed (as adults), incubated, or prefed for 72 hours on a cow. The colonies occurred in 5 different morphologic types (1 to 5) that were similar to those described previously for a Virginia isolate of A marginale in Dermacentor andersoni. The colony density (number of colonies/0.001 mm2 midgut tissue examined) ranged from 0 to 2.0 and was highest in unfed ticks that were not incubated (mean 0.566). Colonies observed by light microscopy were sectioned for study with the electron microscope. The colonies contained both electron-dense forms and reticulated forms. The organisms in type 2 and 3 colonies appeared to be attached to one another, and those in type 4 and 5 colonies occurred separately. Small particles were seen within the limiting membrane of some organisms. A few colonies contained a dense matrix and were surrounded by many small electron-dense particles.

Anaplasma↗

Influence of parasitemia level at feeding on development of Anaplasma marginale Theiler in Dermacentor andersoni Stiles.

Four splenectomized dairy calves were inoculated with a Virginia isolate of Anaplasma marginale Theiler and served as an infective source (donor) for laboratory-reared Dermacentor andersoni Stiles nymphs. Two donor calves developed higher parasitemias during tick feeding than did the 2 other donor calves. One month after molting, adult ticks were incubated at 37 C for 2.5 days to stimulate development of colonies of A marginale, and homogenates of gut were made from ticks fed on each donor calf. Eight susceptible, splenectomized dairy calves (2 per donor calf) were each inoculated IV with the gut homogenates collected from 50 adult ticks and were monitored for patent A marginale infection. Gut tissues from these same groups of ticks were processed for histologic studies. Homogenates from ticks infected on donor calves with lower parasitemias caused infections with longer prepatent periods (av, 43.25 days) than did similar homogenates made from ticks infected by feeding on donor calves with higher parasitemias (av, 31.0 days). Also, sections of gut tissues from ticks infected at the lower parasitemias contained fewer (av, 0.65) colonies/0.001 mm2 of tissue examined than did those from ticks infected at higher parasitemias (av, 6.31).

Anaplasma↗

Immunocytochemical labeling of Anaplasma marginale Theiler in Dermacentor andersoni Stiles with peroxidase- antiperoxidase technique.

Colonies of Anaplasma marginale in midgut epithelial cells of adult ticks that had been infected as nymphs were specifically labeled, using the unlabeled antibody peroxidase-antiperoxidase method of immunocytochemistry. Visual comparison of infected and control tissue sections with the electron microscope demonstrated deposition of ring-like peroxidase-antiperoxidase complexes over organisms within the colonies. The intensity of labeling differed among organisms within a single colony, possibly as a result of varying antigenicity. The labeling observed on organisms in the colonies was similar to that seen on anaplasmal initial bodies in inclusions of infected bovine erythrocytes examined concurrently.

Anaplasma↗

Morphologic characteristics of colonies of Anaplasma marginale Theiler in midgut epithelial cells of Dermacentor andersoni Stiles.

Colonies of Anaplasma marginale Theiler were studied in midgut epithelial cells of adult Dermacentor andersoni Stiles that were infected as nymphs. Colonies were categorized by light microscopy into 5 morphologic types that appear to represent stages in a developmental sequence. The mean diameter of the colony types ranged from 5.64 micrometer to type 1 colonies to 10.49 micrometers of type 5 colonies. Representatives of each colony type were selected by light microscopy and were sectioned for examination by electron microscopy. The morphologic features of individual A marginale organisms within colony types varied and included: (i) small electron-dense forms, (ii) larger reticulated forms, (iii) pleomorphic reticulated forms, and (iv) small particles. Types 1 and 2 colonies contained small, electron-dense forms. Type 3 colonies contained electron-dense forms, early reticulated forms, and small particles that were often outside of the parasite's limiting membrane. Type 4 colonies contained many reticulated forms that often had small particles within cell membranes. Type 5 colonies contained fewer well-formed reticulated forms than did type 4 and, in some cases, large masses of pleomorphic reticulated forms.

Anaplasma↗

Influence of increased temperature on Anaplasma marginale Theiler in the gut of Dermacentor andersoni stiles.

Three splenectomized dairy calves were inoculated with a Virginia isolate of Anaplasma marginale Theiler and served as an infective source for laboratory-reared Dermacentor andersoni Stiles nymphs. One month after molting, groups of adult ticks were incubated at 37 C for 0, 1.5, 2.5, and 7 days. Gut homogenates were made from ticks representing each incubation period. Twenty-four susceptible, splenectomized dairy calves were each inoculated IV with a gut homogenate extracted from 50 adult ticks and monitored for patent A marginale infection. The prepatent periods were determined and used as a measure of infectivity. Gut homogenates made from ticks that had been incubated for 2.5 days produced infections with the shortest prepatent periods, an average of 28.5 days among 3 trials.

Anaplasma↗

Staining characteristics of colonies of Anaplasma marginale Theiler in Dermacentor andersoni Stiles.

Colonies of Anaplasma marginale Theiler were studied in midgut epithelial cells of adult Dermacentor andersoni Stiles that were infected as nymphs. The colonies were demonstrated in plastic sections (1 micrometer) and frozen sections (6 micrometer) with Mallory's stain. Colonies were also demonstrated in frozen sections, using direct fluorescent antibody technique. Colonies were not observed in plastic or frozen sections stained by the Giménez method. In all preparations, the colonies were usually found near the basement membrane of the midgut tissue.

Anaplasma↗

Transmission of Anaplasma marginale Theiler by Dermacentor andersoni Stiles and Dermacentor variabilis (Say).

Cattle were inoculated with a Virginia isolate of Anaplasma marginale Theiler and served as an infective source for laboratory-reared Dermacentor andersoni Stiles and D variabilis (Say) nymphs. Following a molt and subsequent feeding of adult ticks on susceptible cows, transstadial transmission of A marginale occurred from cattle with parasitemias ranging from undetectable (in a carrier cow) to a peak of 27% (in an acutely ill cow). Homogenates of gut tissue from unfed, incubated, and feeding adult ticks that were infected as nymphs were all infective; however, the prepatent period varied according to treatment of ticks after they had molted to the adult stage. Prepatent periods of infections in calves resulting from injection of homogenated tissue from unfed adult ticks was 29 and 34 days longer than the prepatent periods that developed in calves exposed to homogenated tissue from ticks that were feeding or incubated at 37 C for 3 days. Transovarial transmission did not occur when F1 larvae, hatched from eggs produced by female ticks known to be infective, fed on susceptible cattle.

Anaplasmosis↗

Ultrastructure of ANaplasma marginale Theiler in Dermacentor andersoni Stiles and Dermacentor variabilis (Say).

Cattle were inoculated with a Virginia isolate of Anaplasma marginale Theiler and served as an infective source for laboratory-reared Dermacentor andersoni Stiles and D variabilis (Say) nymphs. Anaplasma marginale was demonstrated by electron microscopy in gut tissues of replete nymphal ticks and in unfed, incubated, and feeding adult ticks that were exposed to the organism as nymphs when they fed on an infected cow. The A marginale organism in replete nymphs and adult feeding ticks were morphologically similar to A marginale described previously from infected bovine erythrocytes. Colonies of A marginale were demonstrated by light and electron microscopy in midgut epithelial cells of unfed adult D andersoni and D variabilis that had been incubated at 37 C for 3 days. Anaplasma marginale organisms in colonies were very pleomorphic. Small electron-dense particles were demonstrated in all infected tick stages studied, but were most evident in colonies from incubated, unfed ticks. They may represent a reproductive form of A marginale.

Anaplasma↗

Demonstration of Anaplasma marginale Theiler in Dermacentor variabilis (Say) by ferritin-conjugated antibody technique.

Cattle were inoculated with a Virginia isolate of Anaplasma marginale Theiler and served as an infective source for laboratory-reared Dermacentor variabilis (Say) nymphs. Transstadial transmission of A marginale was demonstrated by feeding the newly molted adult ticks on susceptible cattle and by inoculation of gut homogenates collected from adult ticks at postattachment day 6. A similar gut homogenate from the same group of ticks was caused to react with A marginale-bovine antisera that had been conjugated with ferritin. The homogenate contained organisms similar to those observed in the gut of adult feeding ticks that were infected as nymphs and the labeling of the outer membrane confirmed that organisms observed were A marginale. A gut homogenate prepared from control ticks did not cause infection when inoculated into a susceptible cow and no organisms were demonstrated.

Anaplasma↗

Influence of Ostertagia ostertagi and Cooperia infections on the energetic efficiency of steers fed a concentrate ration.

Studies on effects of subclinical parasitism (low worm burden) in full-fed and maintenance-fed steers indicated that over a short interval low worm burdens do not have a statistically significant effect on the efficiency of energy utilization. In full-fed steers, energy retention tended to be higher in steers with lower worm burdens (mean 95.8 Ostertagia, 12.5 Cooperia) than in those with higher worm burdens (mean 677.5 Ostertagia, 201.8 Cooperia). Maintenance-fed steers generally had larger numbers of Ostertagia ostertagi and Cooperia spp than did the full-fed steers.

Animal Feed↗