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.

Ixodes ricinus ticks are an important vector and reservoir of pathogenic microorganisms causing dangerous infectious diseases in humans and animals. The presence of ticks in urban greenery is a particularly important public health concern due to the potential for humans and companion animals to be exposed to tick-borne diseases there. The study assessed the prevalence of Borrelia burgdorferi and Anaplasma phagocytophilum infection in I. ricinus ticks feeding on dogs.

Investigation was carried out in response to a report of mortality among Jersey crossbred dairy cattle at Therkumedu village, Tirunelveli district which were purchased from Chittoor district, Andhra Pradesh. Clinical examination was carried out in 38 animals. Most of the animals showed signs of general weakness, dullness, anaemia, icteric mucus membrane, lymphadenopathy, acariasis, bruxism, debility and passing semisolid faeces  coated with mucus. Blood smear examination revealed the presence of T. annulata piroplasm and A. marginale inclusions. All the affected animals were treated with Buparvaquone @ 2.5 mg/kg and Oxytetracycline @ 20 mg/kg for 7 days supported with parenteral haematinics (Tribivet) and liver extract. Control of ectoparasites was effected with cypermethrin @ 0.2 per cent as external application and 0.4 per cent spray in animal sheds. All the treated  animals recovered slowly and became normal.

Investigation was carried out in response to a report of mortality among Jersey crossbred dairy cattle at Therkumedu village, Tirunelveli district which were purchased from Chittoor district, Andhra Pradesh. Clinical examination was carried out in 38 animals. Most of the animals showed signs of general weakness, dullness, anaemia, icteric mucus membrane, lymphadenopathy, acariasis, bruxism, debility and passing semisolid faeces  coated with mucus. Blood smear examination revealed the presence of T. annulata piroplasm and A. marginale inclusions. All the affected animals were treated with Buparvaquone @ 2.5 mg/kg and Oxytetracycline @ 20 mg/kg for 7 days supported with parenteral haematinics (Tribivet) and liver extract. Control of ectoparasites was effected with cypermethrin @ 0.2 per cent as external application and 0.4 per cent spray in animal sheds. All the treated  animals recovered slowly and became normal.

A total of 2803 bloodsamples from ruminant livestock werereceived for diagnosis for parasiticinfections from 2011 to 2015. Theruminant samples were from small holderand commercial farms from the states ofNegeri Sembilan, Melaka, Selangor andthe Federal Territory. The results indicatethe presence of theileriosis, anaplasmosis,babesiosis and trypanosomiasis in theseanimals. The most common parasiteidentified annually is theileriosis in cattle.This information is vital for disease controlstrategies as most blood protozoa causemorbidity and is transmitted by vectorssuch as ticks.

Development of the rickettsia, Anaplasma marginale, salivary glands of male Dermacentor andersoni exposed as nymphs or adult ticks, was studied indirectly by inoculation of susceptible calves with homogenates and directly by examination, using light microscopy and a DNA probe; some unfed ticks were incubated before tissues were collected. Salivary gland homogenates made from ticks in every treatment group caused anaplasmosis when injected into susceptible calves; prepatent periods decreased as the time that ticks had fed increased. Colonies of A marginale were seen only in salivary glands of ticks exposed as adults and not in those exposed as nymphs; the percentage of salivary gland acini infected in these ticks increased linearly with feeding time. However, the probe detected A marginale DNA in salivary glands of ticks from both groups; the amount of DNA detected increased as feeding time was extended. The amount of A marginale DNA appeared to remain constant in gut tissues, but to increase in salivary glands. Salivary glands of adult-infected male ticks that were incubated, but did not feed a second time, became infected with A marginale, and the pattern of infection of acini varied with incubation temperature. Development of A marginale in salivary glands appears to be coordinated with the tick feeding cycle; highest infection rate was observed in ticks exposed as adults.

Isolated Anaplasma marginale initial bodies were successfully used in a dot ELISA for rapid detection of antibodies to Anaplasma organisms. The enzyme immunoassay used only 25 ng of antigen dotted onto nitrocellulose disks. Antigen-antibody complexes were detected by use of alkaline phosphatase-conjugated protein A, and reactions were read visually after addition of a precipitable, chromogenic substrate. The test allowed the processing of multiple sera, either for screening or for titer determination, in < 3 hours and was found to be as sensitive as the indirect fluorescent antibody test. The overall performance of the dot ELISA, using isolated A marginale initial bodies, for 580 bovine serum samples was as follows: sensitivity, 93%; specificity, 96%; and predictive value, 95%. Cross-reactivity was not observed with sera positive to Babesia bovis and B bigemina, Trypanosoma vivax, or common bacteria or viruses infecting cattle. The antigen dotted onto nitrocellulose disks was stable when stored at -20, 4, or 25 C. Compared with the indirect fluorescent antibody test, the dot ELISA allowed easier, faster, and more objective interpretation of results. Its simplicity and low cost combined with high sensitivity and specificity indicate that this assay could effectively replace serologic assays currently used for diagnosis of anaplasmosis in cattle.