Genotyping Of Echinococcus Granulosus And Its Comparative Prevalance In Camels And Human Beings
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Publisher: 2008 Dissertation note: Hydaiidosis is caused by metacestode of the dog worm Echinococcus granulosus. It is a serious problem br both Public health and livestock economy. Echinococcus granaiiosu.s has number of genetically distinct strains which are known to differ morphologically and epidemiologically. Out of 100 camels examined only 25 Samples of hydatid cysts were collected from different organs i.e. livers, kidneys, lungs and hearts from Lahore abattoirs. Fertility and viability of the cysts was observed microscopically. Genotyp ing of Echinococcus granulosus was performed through Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP). Seroprevalence of hydatidosis in 25 butchers working in abattoir was also determined by the use of Latex agglutination test (LAT) kit for detection ob hydatidosis.
Considerable information is available about genetic variants of E. granulosus around the world. Ten genotypes of E. granulosus have been described, which exhibit a diversity of morphology, development, and host range, as contrmed by various studies. In the Mediterranean area, the CI or common sheep strain, G2, Camel strain G6, and the equine strain G4 have been found in Spain, Italy, Lebanon, and Syria
To date, molecular studies using mainly DNA sequences have identitied G-6 strain of E. granulosus. This categorization follows very closely the patterns of strain variation emerging from biological and epiderniological traits.
In this study we perform serum analysis of butchers to detect antibodies against Echinococcus so that the prevalence of Echinococcus can be checked; the data available indicated that 14% of butcher's population is infected with Echinococcus. In order to confirm the strain of Echinococcus in camels the PCR-RFLP analysis were performed. The data obtained was analysed and it was concluded that the G6 strain of Echinococciis is prevalent in camels in Pakistan. The results demonstrated that PCRRFLP analysis of samples of patients suspected for Echinococcus is a promising diagnostic method and also confirms the type of Echinococcus prevalent in that area and also enables an early direct detection of parasite DNA. This will help to curtail this drastic malady at an early stage and will help to devise the strategy to minimize the losses due to this disease.
It is hoped that the findings of the present study will be helpful for further planning about the control of the disease and correlating the prevalence in camels and butchers from the zoonotic point of view.
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Molecular Detection And Speciation Of The Canme Piropiasm
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Publisher: 2008 Dissertation note: An epidemiological study of babesiosis in dogs was conducted at Pet center, University of Veterinary and Animal Sciences, Lahore, for one year and information on age, sex and breed was gathered. It was found that from a total number of 6204, dogs up to two years of age were more susceptible than other age groups (2-4, 4-6 and above 6 years).The data regarding genders revealed that males were more prone to the disease than female dogs. As far as the breeds were concerned, crossbred dogs were more susceptible followed by Pointers, Alsatians, German shepherds and Bull terriors.Hot and humid months (June to September) have greater impact on the occurrence of disease. The study regarding identification of ticks revealed that Rhiphicephalus sanguinus is the predominant vector of the disease in Pakistan.
Molecular studies were conducted to characterize and identify the species responsible for canine babesiosis in Pakistan. In this regard, a nested polymerase chain reaction-Restriction fragment length polymorphism (PCR-RFLP) technique was employed on different specimens (Blood, Body tissues and Ticks). For this purpose blood samples were collected from twenty four chronically infected dogs and applied on the Flinders Technologies Associates (FTA) cards for transportation to Australia. Different body tissues (Liver, Spleen, Kidney, Intestine, Bone marrow and Pancreas) were procured after euthanizing the two dogs and DNA was extracted, for further studies. Similarly, the eighty eight ticks were also collected from the infested dogs in the 70% ethanol for transportation to Australia. A nested PCR-RFLP assay was used for the detection and differentiation of Piroplasm species on the basis of the 1 8S ribosomal RNA gene. The assay potentially amplified and identified Babesia gibsoni as the main canine piroplasm. Similar assays on the DNA extracted from body tissues and ticks revealed Babesia gibsoni as the main piroplasm. The PCR was found to have a high detection limit (equivalent to i0 dilution), when using the DNA extracted from blood applied to FTA cards, body tissues and ticks. A new technique was developed for extraction of DNA from FTA cards and tick, in this technique, instead of using the FTA specified punching machine, we used scalpel blades, and so the rest of the chemicals used are'generally and easily available. The same protocol was used for extraction of DNA from ticks, only chemicals used in different quantities with different spinning times. Both of which, resulted in cost reduction, less effort and speedy DNA extraction. The technique reported here has the potential to be standardized for routine DNA extractions from FTA cards and ticks.
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Epidemiology, Molecular Diagnosis And Chemotherapy Of Giardiasis In Bovine
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Publisher: 2009 Dissertation note: Giardia is a protozoan parasite of the small intestine that causes extensive morbidity worldwide. Dairy calves can excrete high numbers of the cysts of Giardia and the disease in cattle is clinically important and can reduce the growth performance of the ruminants. Giardia is the cause of non-viral diarrhoea in humans and is responsible for epidemics in the developed and developing countries. The cyst is the infectious form, is ingested in contaminated water or food or directly from faecal-oral contact. Giardia duodenal is the only species, which is found in both humans and animals including dogs, cats, bovines, pigs, sheep and equine.
The present study was conducted to determine the prevalence in bovines at Military dairy farm, Gawala dairy colonies, the Government dairy farm and Household dairies in Lahore. The effect of season, sex, and age on infection rate and shedding of the cysts were also noted, and association of the Giardia infection with normal and abnormal stools was also studied.
Overall 2160 bovine faecal samples (720 buffaloes, 720 cattle and 720 calves) were examined during the study period from August 2007 to July 2008, amongst calves 362/720 (50.27%) were found to be positive. The highest prevalence was recorded in the Government. Dairy farm (68.33%) followed by Gawala colonies (55%), then the Military dairy farm (44.33%) and the lowest (34.44%) was recorded in Household dairies. Overall, highest (61.6%) seasonal prevalence was recorded during autumn, followed by spring (60.83%), then summer (53.4%) and the lowest
(34.1%) was recorded during winter. The highest (65%) prevalence was reported during August and the lowest (3 0%) during December.
Females were found to be more susceptible (56.74%) than males (35.1%). The prevalence was significantly higher (71.52%) in younger calves than the adults
Overall prevalence in cattle was 28.05%. The highest (41.67%) prevalence was recorded at the Government dairy farm, followed by Gawala colonies (32.72%), then the Military dairy farm (22.72%) and the lowest (15%) was recorded in Household dairies. The highest (35%) prevalence was found during August and the lowest (21%) during January. A significant difference (P<0.05) was noted. Females were found to be more susceptible (29.21%) than males (18.75%). The young calves had significantly higher (3 8.88%) prevalence as compared to the adults (24.44%).
Similarly, the overall prevalence in buffaloes was found to be 20.11% percent. The highest (40.55 %), prevalence was recorded at the Government Dairy Farm, followed by Gawala colonies (30%) then Military Dairy Farm (21.11%) and the lowest prevalence i.e. 12.77% was reported in Household Dairies. A non significant difference was recorded P>0.05). The highest (46.66 %) prevalence was recorded during August, while, the lowest (6.66%) during November and December. Females were found to be more susceptible than males. Where as the prevalence in a younger buffalo was significantly higher as compared to the adults.
Comparison of direct microscopic examination and PCR based methods was made at the Government dairy Farm, Gawala colonies; Military Dairy Farm and
Household Dairies. By direct Microscopic examination prevalence was found to be 28.05% (202/720) in cattle whereas by PCR it was 31.11%. Statistically analysis showed that the prevalence by PCR was significantly (P<0.05) higher than the microscopic examination.
It was observed that the highest prevalence of Giardiasis in bovines (Calves, Cattle and buffalo) was noted during August when the average temperature was 31.48°C. However the maximum and minimum temperatures were 35.37°C and
27.6°C, relative humidity 7 1.28% and rainfall was 3.2mm. The results of therapeutic trials by using albendazole, metronidazole, and mebendazole in cattle were calculated on the basis of reduction in the cysts count in the faeces after treatment.
Efficacy of albendazole at three dose levels i.e. 1 Omg/kg.b.wt, 1 5mg/kg.b.wt, 2Omg/kg.b.wt was 86.33%, 98.5% and 100% respectively, on day 27 after treatment. Efficacy of the metronidazole at 5Omg/kg.b.wt, 1 OOmg/kg.b.wt, and 1 5Omg/kg.b.wt. Was 85.42%, 87.8% and 94.02% respectively on day 27. Efficacy of mebendazole at three dosage level i.e. 7.5rng/kg.b.wt, lOmg/kg.b.wt and 2Omg/kg.b.wt was 81.15 %, 87.32%, and 90.4% on day 27 after treatment. Among these drugs, albendazole at
1 5mg/kg.body.weight was found to be most effective drug in the elimination Giardia infection. The significant (P<0.05) decrease in the CPG count after treatment in all the three groups and dose levels was noted. A significant difference (P<0.05) was observed in the level of leukocytes and of eosinophisl of infected cattle at day 06 and day 13 post inoculation. The leukocytes/lymphocytes count of Giardia infected cattle was 58.09%. Whereas, eosinophils constituted of leukocytes 9.69%. The total proteins of the sample were studied by sodium doedocyl sulphate polyacrylamide gel ELECTROPHORESIS (SDS PAGE). The result indicated that 8 diffeent molecular weight peptide badns were identified with size ranges from 20 to 70 KDa and common bands reported at 20, 24 and 35 K Da
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Studies On Rats And Mice As A Reservoir Of Zoonotic Parasites
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Publisher: 2009 Dissertation note: A total of 3600 rats and mice were examined for the prevalence of various zoonotic parasitic infections. Species of various parasites were identified. The order of ifection rate was as Hymenolepis nana (59.9%), Toxoplasma gondii in rats and mice 52.0%), Hymenolepis diminula (35.4%), Cryptosporidium parvum (16.5%), Taenia taenieformis (11.7%), Toxoplasma gondii in humans (11.3%), Capillaria hepatica 7.0%), Babesia spp. (2.3%), Trypanosomu spp. (2.3%) and Leishmania spp. (1.2%).
The highest (78.0%) month-wise prevalence of FL nana was found during August whereas the lowest (43.0%) during February. The highest (70.8%) season-wise prevalence was noted during summer followed by autumn (62.9%) then spring whiles the lowest (46.8%) during winter. Infection was higher in males (78.0%) than females 45.9%). Adult rats and mice were more commonly affected than younger. Infection was higher in rats (65.1%) than mice (16.3%).
The highest (48.0%) month-wise prevalence of H diminuta was found during August whereas the lowest (28.0%) during January. The highest (45.4%) season-wise prevalence was noted during summer followed by spring (35.1%) while the lowest (29.3%) during winter. Infection was higher in males (43.8%) than females (29.3%). Adult rats and mice were more commonly affected than younger. Infection was higher in rats (37.3%) than mice (20.2%).
The highest (23.0%) month-wise prevalence of C. parvum was found during July whereas the lowest (10.0%) during December. The highest (22.4%) season-wise valence was noted during summer followed by spring (16.9%) then autumn (15.3%) le the lowest (11.3%) during winter. Infection was higher in females (19.9%) than es (12.0%). Adult rats and mice were more commonly affected than younger, infection was higher in rats (17.7%) than mice (7.6%).
The highest (20.0%) month-wise prevalence of T raenieformis was found during y and August whereas the lowest (6.0%) during December. The highest (18.4%) son-wise prevalence was noted during summer followed by autumn (10.8%) then ring (10.6%) whereas lowest (7.0%) during winter. Infection was higher in females 15.6%) than males (6.5%). Adult rats and mice were more commonly infected than younger. Infection was higher in rats (12.4%) than mice (6.0%).
The highest (12.7%) month-wise prevalence of C. hepatica was found during August whereas the lowest (4.0%) during November. The highest (10.8%) season-wise prevalence was noted during summer followed by autumn (6.0%) then spring (6.2%) hues the lowest (4.9%) during winter. Infection was higher in females (8.4%) than males (5.0%). Adult rats and mice were more commonly affected than younger. Infection higher in rats (7.3%) than mice (4.6%).
The highest (9.0%) month-wise prevalence of Babesia spp. was found during August whereas infection was absent during January, February, March, April, November and December. The highest (5.0%) season-wise prevalence was noted during summer followed by autumn (4.2%) then spring (0.1%) and was absent during winter. The infection was higher in males (2.7%) than females (2.0%).The infection was present only in adult rats and mice and no case was recorded in younger. Higher infection was noted -n (2.3%) and than mice (2.4%).
The highest (7.3%) month-wise prevalence of Trypanosoma spp. was found August whereas it was absent during January, February, November and December. The highest (5.7%) season-wise prevalence was noted during summer by spring (2.2%) then autumn (1.2%) and it was absent during winter. The infection was higher in females (2.5%) than males (2.0%). Adult rats and mice were more commonly affected than younger. Higher infection was noted in rats (2.5%) than mice
The highest (4.7%) month-wise prevalence of Leishmania spp was found during y and August whereas infection was absent during January, February, March, April, October, November and December. The highest (3.4%) season-wise prevalence was during Summer followed by autumn (1.1%) then spring (0.1%) and infection was during winter. The infection was higher in females (1 .4%) than males (0.9%). The infection was found only in adult rats and mice and no positive case was recorded in 1younger. The infection was (1.2%) in rats and mice.
The role of meteorological data i.e temperature, humidity and rainfall was also studied. It was noted that there was a positive co-relation of high temperature and humidity with the prevalence of various parasitic infections. As the infection were maximum during summer when the temperature and humidity was favorable for high prevalence of parasitic burden.
This study will he helpful to disseminate information regarding zoonotic potential parasitic infections via media i.e. newspapers. magazine, electronic media (T.V, I.T, Radio) through seminars, meetings and conferences with professionals doctors, paramedical staff and livestock farmers.
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Epidemiology, Serodiagnosis, Therapy And Control Of Schistosomiasis In Buffloes
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Publisher: 2008 Dissertation note: Schistosomiasis is one of the major diseases of public health and socio-economic importance in the developing countries of the world. Among human parasitic diseases, Schistosorniasis ranks second to malaria in tern-is of world wide public health risk. Keeping in view the importance of disease, the study was conducted to record the month wise and season wise prevalence of Schistosorniasis in buffaloes in four districts of the Punjab, province ie., Lahore, Sargodha, Kasur and Sheikhupura. The present study comprises of four parts.
Part I: deals with epidemiology of Schistosomiasis in buffaloes. Overall prevalence of Schistosomiasis in buffaloes, different farms of Punjab province indicated that infection was the highest (17%) at Kasur district followed by Sheikhupura (14.83), then Lahore (14.6%) and the lowest (13.66%) at Sargodha. The highest month wise prevalence was recorded during August (25.5%) followed by July where as the lowest during December and January.
Infection in buffaloes was higher in animals over two years of age (1 9%) than animals below two years of age (5%) in all the four districts of Punjab. The prevalence was higher in females (15.98%) than male (9.48%). There is variation in the prevalence as there is difference in the environmental and managemental condition of the area. For the serodiagnosis i.e. ELISA was used, the results indicated that the prevalence was lesser than the faecal examination because this was more specific and sensitive than the faecal examination.
Part 2: deals with the prevalence and ecology of snails. Various species of snails which act as the intermediate host of the Schistosomes were collected from the study area. The e of infection in the snails and role of cercariae in transmission of the disease was studied. A total of 10418 snails were collected of these 13.51 per cent were found to be infected. Among these 2350 were collected from Kasur district with infection rate of 14.51 percent followed, by Sheikhupura 2882 (13.6%) then Sargodha 2709 (13.40%) and the lowest at Lahore 2477 (12.51%). At Kasur district, genus wise prevalence of snails with infection rate indicated that Oncomelonia, indoplanorbis and Bullinus are the predominant genera with infection rate of 31.79, 17.10 and 14.46 percent respectively. However the highest number of the snails collected belonging to the genera Indoplanorbis. At Sheikhupura district, genus wide prevalence of snail indicated that Bullinus, Lymnaea, Indoplanorbis and Physa are the four prominent snails with infection rate of 24.74, 20.57, 14.66 and 13.84 percent respectively. At Sargodha district, genus wise prevalence of snails indicated that Lymnaea, Indoplanorbis, Bullinus and Physa are the four prominent snails with infection rate of 25.09, 14.29, 14.28 and 16.77 percent respectively. At Lahore district, genus wise prevalence of snails indicated that Bullinus Lymnaea, Physa and Indoplanorbis are the four prominent snails with infection rate of 23.37, 18.96, 13.97 and 12.70 percent respectively.While the prevalence at the snail level the Chi square value is 242.944 and the P-Value is 0.0000 1 which is highly significant.
Part 3: deals with the meteorological data ie, temperature, humidity, rainfall and pan evaporation with prevalence of snails and parasites. The temperature and rain fall play very important role in the spread of disease. The ideal temperature ranges form 22-25 °C where development within snail takes place in an efficient manner similarly humidity
f ranges from 55-70% is ideal for the development of the snail and the parasite. Rainfall is very important for the spread of the disease. There is a positive correlation of disease incidence to maximum and minimum temperature, humidity, and rainfall and pan evaporation. It was seen that during summer and autumn, optimum temperature, relative
humidity and rainfall play an important role for rapid propagation of the parasitic life
Part 4: deals with therapeutic trials against Schistosomiasis in buffaloes. A total of 150 animals (140 infected and 10 animals, normal) age ranged 5-9 years and of both sexes naturally infected with Schistosorniasis were used in thirteen controlled experiments. The efficacy of certain indigenous drugs, including Nigella sativa (Kalongi) , Caesalpinia
Crista (Karangwa), Lagenaria siceraria seeds (Kadoo ke Beej), Sausseria lappa (Qushte-e-Shreen) and Praziquanlel was compared with each other and control. Efficacy was quantified by determining the difference of egg per gram faeces (EPG) pre and post treatment. After the single dose of 50, 75 and 100 mg 1kg body weight of Nigella sativa (Kalongi) reduced EPG by 65.85, 68.29 and 71.79 per cent, respectively. After the second dose the respective reduction in EPG was 85.36, 92.68 and 94.87 percent. Caesalpenia crista at three dosage levels i.e.50, 75 and 100 mgI kg body weight caused 46.34, 53.65 and 59.52 percent reduction respectively while the reduction in EPO after second dose was 82.92, 90.24 and 92.85 percent respectively. Lagenaria siceraria Seed at three dosage levels i. e., 50, 100 and 150 mg/ kg body weight caused the reduction in EPG reduction 47.61, 52.63 and 64.10 percent respectively, while after second dose, counts as the reduction 80.95, 86.84 and 92.30 percent respectively. Sausseria lappa at three dosage levels i.e., 100, 150 and 200 mg/ kg body caused EPG reduction as under 50.00, 53.48 and 56.09 percent respectively, while after second dose the reduction in EPG count was 71.42, 81.39 and 85.36 percent respectively. Where as Praziquantel at the dose of 10 mg/body weight caused reduction in EPO 66.66% while after the second dose the reduction in EPG count was 97.43 percent. The efficacy order was Praziquantel, Nigella saliva, Caesalpinia crista, Lagenaria siceraria and Sausseria lappa. No side effects with any drug were noted. All animals showed clinical improvement after the treatment.
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