Anaplasma marginale, morphologic characteristics of colonies in Dermacentor andersoni midgut epithelial cells

Theileria mutans, 4 isolates obtained from geographically distant parts of South Africa by subinoculation of infected blood were readily transmitted to cattle by Amblyomma hebraeum, all attempts to transmit these isolates with Rhipicephalus appendiculatus were unsuccessful

Anaplasma marginale, transstadial transmission by Rhipicephalus simus, successful transmission to oxen by tick stabilate prepared from infected male ticks

Objective--To evaluate the efficacy of coumaphos, an organophosphate (OP) acaricide, at concentrations up to 2 times higher than the highest concentration required by the US Eradication Program against all stages of an OP-resistant strain of Boophilus microplusin experimentally infested cattle. Animals--16 tick-naïve 200-kg female Hereford calves. Procedure--Four groups of cattle (4 calves/group) were all infested with Boophilus ticks 3 times before treatment. Each group was treated with coumaphos as follows: group 1, at 0.165% active ingredient (AI); group 2, at 0.299% AI; group 3, at 0.566% AI; and group 4, not treated. Following treatment, ticks were collected for 21 days. Ticks collected 1 to 7, 8 to 14, and 15 to 21 days after treatment were considered adults, nymphs, and larvae, respectively, at time of treatment. Results--Overall control at 0.165, 0.299, and 0.566% AI was 52.9, 75.8, and 89.7%, respectively. Control of adults ranged from 4.3% at 0.165% AI to 73.5% at 0.566% AI. Control of nymphs ranged from 60.6% at 0.165% AI to 97.3% at 0.566% AI. Control of larvae was > 98% at all coumaphos concentrations. Conclusions and Clinical Relevance--All coumaphos concentrations failed to provide acceptable control for use in the US Eradication Program against OPresistant ticks. Treatment was least effective against adults and most effective against larvae. Even at 0.566% AI (2 times higher than required by the US Eradication Program), ticks were not eradicated, placing the United States at risk from dispersing cattle harboring viable ticks to uninfested areas.

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

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.

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.

influence of parasitemia level of calves at time of feeding by Dermacentor andersonii upon development of Anaplasma marginale in the ticks

Boophilus microplus engorged females stored at 4 C for up to 5 days and females kept at room temperature for 1 day and then at 4 C for 1 day showed no significant differences in their response to an organophosphate (dioxathion) as determined by the Drummond adult test

Rhipicephalus sanguineus and Ctenocephalides spp. on kennelled dogs, seasonal prevalence and mean numbers and percentages of immature and adult ticks and of fleas recovered from various sites on dogs