The field study was carried out in indigenous and cross bred cattle in three districts of Punjab namely, Lahore, Multan and Rawalpindi during the year from September 2006 to August 2007. A total of 1200 cattle were selected as reference population comprising of 600 Sahiwal and 600 Cross bred animals. 300 blood samples were collected during each season by taking 100 samples from each district. The results of field study on the basis of PCR showed that prevalence of Tannulata during summer was highest 46.7% in both breeds of cattle while lowest prevalence was recorded in winter, 4.6 % and autumn season 4.3% . The breed wise prevalence of Tannulata was lower, 11.5 % in sahiwal cattle compared with 20.2 % in cross bred cattle. The mortality rate of 2.8% was noticed only in cross bred cattle with highest mortality recorded in district Rawalpindi ,64.7% and lowest in district Multan,29.4% .The positive percentage of Tannulata was higher,61.5%(234/380) in females as compared to males ,38.4% (146/3 80) of both breeds. The positive percentage in much animals was highest, 32.4 %( 123/380) in both breeds while lowest was recorded in heifers, 11.1 %( 42/3 80). In males of both breeds 26.3% (100/3 80) positive percentage was seen in adults above 1 year of age while lower, 12.1% (46/3 80) was recorded in young animals up to 1 year of age. The prevalence of Tannulata was highest,l2.2% in district Lahore in both breeds while lowest prevalence, 9.33% was seen in district Rawalpindi.

The efficacy of PCR test was highest ,31.6% followed by microscopic lymph node smear examination,8.25% and microscopic blood smear examination,6% in diagnosis of field challenges of Theileria.

During the present study mixed infection with Babesia bigemina, 158%, was recorded in reference population. TLc breed wise prevalence of Babesia bigen'iina was higher in cross bred animals 33.33 % compared to 17 % in sahiwal cattle respectively along with 6% prevalence of the ileria specie on blood smear examination.

The overall economic losses of 3.39 million were calculated during the present study in three districts under study. The stocking pattern was highest (47%) for herd size of 1-2 animals while lowest herd size (12%) of 5-6 and above 6 was recorded.

The survey of ectoparasites showed the highest prevalence of ticks,66.7% recorded in district Lahore while prevalence of lice was highest,36.3% in district Rawalpindi and prevalence of mange mites was highest ,4% in district Multan. The highest prevalence of Hyalomma 12.0%, followed by Boophilus 8.1% Haemaphysalis 5%.and Rhipicephalus 3.1% were recorded. During the present study the species of Hyalomma identified were Hyalomma a. anatolicurn (65.2%) and Hyalomma n-zarginatum marginaturn (34.8%).The examination of salivary glands revealed that Hyalomma a. anatolicum (87%) and Hyalomma marginatum inarginatum (20.8%) were infected with T. annulata sporozoites. It was observed that the population of ticks was heaviest in the month of June, mid September to mid October with lowest infestation during the month of November. No ticks were noticed on animals from December to February.

The taxonomical study of Hylomma species showed the difference between both sexes and in different developmental stages. The two species of Hyalomnia in the present study were differentiated on the basis of structural features in adults. The pre-oviposition & oviposition periods recorded during spring were longest i.e 9 days and 12 days respectively. The incubation period of the ova of Hyalomma in summer and autumn was longest i.e 20 days. Mean survival period of unfed larva of Hyalomma was recorded as 56 days while for nymph it was 65 days. Larval and nymph engorgement period was longest in spring i.e. 9 days and 7days respectively while for adult the mena engorgement period in spring was longest i.e 9 days .The larval and nymph moulting period was longest in spring i.e 16 days and 17 days respectively . Amount of blood sucked in mg by first instar of Hyalomma ranged from 0.132 -0.126mg while for second instar it ranged from 140-79mg and for third instar it was 237-180 mg. The eggs were oblong in shape and measured 0.470 X 0.420 mm in size with weight of 0.041 rng on an average. It was observed that the maximum number of eggs laid by a single female tick in spring varied from 3720 to 3918, in summer from 2611 to 2961 and in autumn from 2423 to 2606.

The bionomical study showed effect of varying temperature and humidity on the development of different stages of Hyalomma tick. The effect of constant temperature (30°C) and varying humidity showed mean pre-oviposition period of mid October with lowest infestation during the month of November. No ticks were noticed on animals from December to February.

The taxonomical study of Hylomma species showed the difference between both sexes and in different developmental stages. The two species of Hyalomnia in the present study were differentiated on the basis of structural features in adults. The pre-oviposition & oviposition periods recorded during spring were longest i.e 9 days and 12 days respectively. The incubation period of the ova of Hyalomma in summer and autumn was longest i.e 20 days. Mean survival period of unfed larva of Hyalomma was recorded as 56 days while for nymph it was 65 days. Larval and nymph engorgement period was longest in spring i.e. 9 days and 7days respectively while for adult the mena engorgement period in spring was longest i.e 9 days .The larval and nymph moulting period was longest in spring i.e 16 days and 17 days respectively . Amount of blood sucked in mg by first instar of Hyalomma ranged from 0.132 -0.126mg while for second instar it ranged from 140-79mg and for third instar it was 237-180 mg. The eggs were oblong in shape and measured 0.470 X 0.420 mm in size with weight of 0.041 rng on an average. It was observed that the maximum number of eggs laid by a single female tick in spring varied from 3720 to 3918, in summer from 2611 to 2961 and in autumn from 2423 to 2606.

The bionomical study showed effect of varying temperature and humidity on the development of different stages of Hyalomma tick. The effect of constant temperature (30°C) and varying humidity showed mean pre-oviposition period of


that the values of TEC, TLC, and PCV and hemoglobin decreased considerably. Comparison of normal average with affected average values increased in Cross bred cattle while for Sahiwal cattle the values increased for lymphocytes and decreased for neutrophils, eosinophils, monocytes and basophils. The experimental study showed highest preyalence of disease in adult animals as compared to young animals by all diagnostic tests. The prevalence of disease in young animals at 12 months of age was highest (85.7% by MLE,71 .4% by MBE,85.7% by IFA & PCR in Sahiwal while 71.4% by MLE & MBE, 89% by IFA, 100% by PCR in cross bred animals. ) while lowest percentage of disease was seen at 3 months of age by all tests.The highest specificity and sensitivity ,96% and 75% respectively for PCR test was recorded while for MBE lowest specificity and sensitivity ,76% and 44% respectively were calculated. PCR analysis of the samples with 6u1 of MgCl2 gave successful results . It was found out that primers set A anneal at Tm 55 °C while Primer set B anneal at 60°C. The expected 721-bp fragment was generated from T. annulata DNA with primer set N5 1 6/N5 17721 -bp liagment with 0.00040% parasitemia, corresponding to 19 parasites per ml while primers 989 and 990 amplified the expected 1098 bp fragment of DNA. All animals that were positive by microscopic examination were also positive by IFA as well as PCR. The therapeutic trials showed efficacy of buparvaquone @ J/M 2.5 mg /kg body weight and Calotropisprocera @ 0.3mg! Kg dose orally.8 doses on alternate days was 60% and 100% respectively. After completion of treatment with Cal otropis procera no tick infestation was seen while ticks were present on the body of that the values of TEC, TLC, and PCV and hemoglobin decreased considerably. Comparison of normal average with affected average values increased in Cross bred cattle while for Sahiwal cattle the values increased for lymphocytes and decreased for neutrophils, eosinophils, monocytes and basophils. The experimental study showed highest preyalence of disease in adult animals as compared to young animals by all diagnostic tests. The prevalence of disease in young animals at 12 months of age was highest (85.7% by MLE,71 .4% by MBE,85.7% by IFA & PCR in Sahiwal while 71.4% by MLE & MBE, 89% by IFA, 100% by PCR in cross bred animals. ) while lowest percentage of disease was seen at 3 months of age by all tests.The highest specificity and sensitivity ,96% and 75% respectively for PCR test was recorded while for MBE lowest specificity and sensitivity ,76% and 44% respectively were calculated. PCR analysis of the samples with 6u1 of MgCl2 gave successful results . It was found out that primers set A anneal at Tm 55 °C while Primer set B anneal at 60°C. The expected 721-bp fragment was generated from T. annulata DNA with primer set N5 1 6/N5 17721 -bp liagment with 0.00040% parasitemia, corresponding to 19 parasites per ml while primers 989 and 990 amplified the expected 1098 bp fragment of DNA. All animals that were positive by microscopic examination were also positive by IFA as well as PCR. The therapeutic trials showed efficacy of buparvaquone @ J/M 2.5 mg /kg body weight and Calotropisprocera @ 0.3mg! Kg dose orally.8 doses on alternate days was 60% and 100% respectively. After completion of treatment with Cal otropis procera no tick infestation was seen while ticks were present on the body of animals treated with buparvaquone. With herbal treatment the animals showed diarrhoea for first 10-12 hours after administration of every dose on alternate days but animals recovered spontaneously without any antidiarrhoeal treatment. The results of CBC showed the characteristic macrocytic hypochrornic anemia in theileriosis was recovered by Calotropis procera treatment. The of result of LFT's and kidney function tests post treatment with Calotropis procera showed no toxicity of drug. In group C the characteristic signs of disease were noticed.

The results of prophylactive trials with both drugs showed delay of 23 days in the onset of clinical disease with buparvaquone while clinical disease was not seen in second group that was prophylactively treated with Calitropis procera. The in vitro trials with both drugs to check the acaricidal activity supported trials with Calitropis procera as having acaricidal within 3 hours while no such effect was noticed with bupasrvaquone in vitro trials.