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1. Molecular Detection And Speciation Of The Canme Piropiasm

by Isma Nazli Bashir | Prof. Dr.Zrafar Iqbal Ch | Dr.Peter J.Irwin | Prof. Dr. Azhar Maqbool | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2008Dissertation 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. Availability: Items available for loan: UVAS Library [Call number: 1061,T] (1).

2. Serofpidemiology, Zoonotic Potential And Chemotherapy Of Neosporosis In Dogs And Cattle

by Muhammad Mudasser Nazir | Prof. Dr. Azhar Maqbool | Dr. Muhammad Lateef | Prof. Dr.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2010Dissertation note: The aim of current study was to demonstrate the most important features of Neospora caninum infection in Pakistan. In the present study, I examined the prevalence of N. caninum in 7 districts of the country and to accessed the efficacy of various drugs against the parasite in cell culture. For the achievement of this purpose, the core objectives were, To have an overview on the overall seroprevalence of neosporosis throughout the country by means of cELISA in aborting, at risk and clinically healthy cows. To check the correlation of Iscom ELISA and cELISA, and determination of prevalence of N. caninum by means of Iscom ELISA on milk samples. To identify the transmission of disease towards human. To determine the efficacy of various drugs against N. caninum. In phase 1, overall seroprevalence of N. caninum in dairy cattle (detected by means of cELISA, VMRD, Inc., Pullman, WA, USA) was found to be 43.4% with a significant difference (P < 0.05) of seropositivity among all 18 herds (n = 5 aborting herds, n = 13 non-aborting/clinically healthy herds) selected from 7 districts of Pakistan. The seropositivity of cattle to N. caninum antibodies was significantly higher in aborting animals (52.7%) as compared to non-aborting cows ( 41.5%), indicating a significant difference between aborting and non-aborting cattle. In case of pregnant and non-pregnant animals, similar findings were recorded in our study. A significantly higher rate of seroprevalence was observed in pregnant dams (59.8%) than non-pregnant cattle (35.2%). Overall, higher serological prevalence was evaluated during the summer season (61.1%) in all areas followed by autumn (46.9%), spring (34.9%) and least seropositivity was observed in winter season (26.6%). The difference in seropositivity was significantly different among all age groups, greater in animals older than 2 years of age. Furthermore, the prevalence was statistically significant (P < 0.05) among cattle of different breeds. Seroprevalence in cases of crossbred animals were higher followed by exotic and indigenous breeds. Phase 2, describes the seroprevalence of N. caninum in clinically healthy dairy cows. A selection of 760 animals from 13 dairy herds located in Punjab and Sindh Province, Pakistan to demonstrate the presence or absence of the Neospora caninum infection in commercial dairy cattle. The serostatus of the cows towards N. caninum was detected by cELISA (VMRD, Pullman, WA). Out of 760 animals, (43.2%) were seropositive to N. caninum. A significant difference of positivity was recorded among all 13 dairy herds. Age wise prevalence though not statistically significant among all age groups, was greater in animals over 2 years of age and least in heifers. Variation was also observed in samples from cattle of different breeds. A significantly higher prevalence was observed in crossbred animals than in purebred and nondescript cattle. Seasonal prevalence was higher during summer season than rest of the seasons. The seroprevalence of N. caninum in pregnant cows was significantly greater than in non-pregnant animals. Second experiment of this phase describes the seroprevalence of 240 animals from 5 herds with a high rate of abortion, the percentage of seropositivity observed in these herds was 43.8%, slightly higher than the clinically healthy and non-aborting cattle. No significant difference was observed among all sample locations in this experiment. However, significant difference of positivity was recorded among different breeds of cattle. Age wise prevalence, though not statistically significant (P > 0.05), was greater in animals older than 2 years of age. The assessment of milk samples from lactating cows were also determined for Neospora caninum antibodies by means of Iscom ELISA (SANOVIR® Sanova Biotech AB, Uppasala, Sweden) and showed a good level of agreement (r² = 0.9959) between the two tests (cELISA and Iscom ELISA). Although, the cELISA (VMRD, Inc., Pullman, USA) expressed a higher seropositivity and sensitivity than Iscom ELISA (Sanova Biotech AB, Uppasala, Sweden). Therefore, both of the ELISA tests (cELISA and Iscom ELISA) for the detection of N. caninum antibodies in dairy cattle can perform better in lactating animals. The Iscom ELISA has some advantages over cELISA as it's easy to collect milk samples than serum samples, moreover Iscom ELISA is cheaper and easy to use but has low sensitivity than cELISA and cannot be used in dry animals. The percentage of positivity detected through Iscom ELISA on individual milk samples were 61.4% and 76.6% by cELISA. In phase 3, an epidemiological study was conducted to determine seroprevalence of N. caninum in dogs of different breeds and age groups. The serum samples of dogs were analyzed by cELISA (VMRD, Pullman, USA) showed a seropositivity of 23.5%. There was no significant difference of seropositivity among various sample locations, highest prevalence was observed in Muzaffar Garh (31.9%), followed by Gujranwala (27.9%), Lahore (25.1%), Hafizabad (20.2%) and least prevalence was recorded in district Okara (14.6%). A significant difference in prevalence of N. caninum antibodies between male (26.1%) and female (18.8%) dogs were recorded. The difference in seroprevalence was not significant among all age groups. The samples with no age record showed a highest prevalence (29.5%) and least seropositivity was observed in adult dogs of 3-6 years of age (18.7%). During Summer season, highest positivity to N. caninum was (31.0%) recorded while the lowest prevalence (16.0%) was observed in Winter season. Phase 4, describes the seroprevalence and transmission of N. caninum in humans. A selection of 52 serum samples from humans was analyzed for the presence of N. caninum antibodies. The serostatus of the humans towards N. caninum antibodies (IgG) was determined by using commercially available antigen coated IFAT slides (VMRD, Inc., Pullman, Washington USA) and human conjugate. Overall very low prevalence (1.9%) of N. caninum antibodies was reported in this study. Only one case was found to be positive, these findings indicate that no strong evidence of N. caninum infection in humans. In phase 5, in vitro drug trials was conducted to access the best efficacy of three commercially available drugs. We found that among three anticoccidial drugs i.e Clindamycin, Diclazuril and Sulfadiazine, Diclazuril has best inhibitory effect against N. caninum tachyzoites in cell culture followed by Clindamycin and sulfadiazine. Availability: Items available for loan: UVAS Library [Call number: 1575,T] (1).

3. Epidemiology Zoonotic Potential Haematology Amd Chemotherapy Of Sarcoptic Mange In Camel In Punjab

by Muhammad Irfan Zahid (2011-VA-800 | Prof. Dr. Azhar Maqbool | Prof. Dr. Muhammad Sarwar Khan | Prof. Dr.Shazia Anjum | Prof. Dr. Kamran Ashraf.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2015Dissertation note: A camel is a very hardy ruminant animal, which can survive under harsh climatic conditions very effectively by utilizing the marginal areas with excellent capabilities and produce under such conditions (Hjort and Hussein, 1986; Abbas and Tilley, 1990). Camel is an important animal as it is well adopted in unique manners in the hot, arid and semi-arid environments (Schwartz, 1992). It can survive without water and food for many days and this unique ability of camel makes it an ideal for such harsh conditions for which it is also commonly known as “The Desert Ship”. In spite of the fact that camel is an important member of a group of animals which produces food for human consumption in the shape of milk and meat, yet it is the most neglected one in the field of scientific research. It may be due to the fact that camel belongs to such areas of the world which are arid, semi-arid or rain fed in nature, having harsh climatic conditions, where poor nutrition and poor management are the major issues (Sohail, 1983). It is an established fact that diseases originating from parasites lead to the main health hazard issues in animals. These parasites survive at the expense of the host animals causing lot of health problems, like skin irritation, anemia leading to weakness and debility. Some of the parasites have zoonotic importance and may become a source for the transfer of many contagious diseases like scabies to the human beings (Dominguez et al. 1978). McClain et al. 2009, observed the scabies as a major health problem globally both for humans and animal population. Sarcoptes scabiei is an ectoparasite which is a cause of scabies, a skin problem in the human beings worldwide and the similar species of mites do also produce a similar type of disease in a large variety of wild and domesticated mammals (Pence and Ueckermann, 2002; Fitzgerald et al. 2004). Fain, 1978, reported that more than fifteen (15) different species of Sarcoptes scabiei morphologically and genetically distinct from each other have been identified in different hosts. Introduction 2 Sarcoptic mange is the second important problematic disease of camel after Trypanosomiasis (Nayel and Abu-Samra, 1986). Scabies caused by Sarcoptes scabiei var cameli is a serious & highly contagious skin problem and also economically important disease of the camels (Pegram and Higgins, 1992). Camels, which are reared with deficient nutrition, poor management and under unhygienic conditions are mostly affected by this disease (Kumar et al. 1992). A large group of people and communities living in arid diverse ecozones in the entire world, particularly in harsh climates earns their livelihoods by depending on camels. This dependence may spread to the utilization of camel milk, meat, wool and leather besides its use in transportation, riding and sports (Wilson, 1984; Snow et al. 1992). In Pakistan camels are also raised by the people for meat, milk, riding, transportation and sports purposes in the deserts, semi desert & rain-fed / warm areas of the entire country being a hardy animal as it can tolerate easily the rugged climate as well as extremes of temperatures of such areas. The natural harsh and adverse climatic conditions, particularly during long dry seasons lead to a paucity of feeding regimes resultantly the camels raised in such areas are subjected to stress conditions which lower their resistance and make them easily vulnerable to diseases (Abbas et al. 1993; Agab, 1993). Abbas & Tilley, 1990; Saint-Martin et al. 1992; Abbas and Agab, 2002; Pathak and Chhabra, 2010; while reviewing the parasites & parasitic diseases of camel population in India were of the opinion that Sarcoptic mange is a serious, debilitating, dreaded and widely prevalent disease of camels in India. Besides other infectious diseases of bacterial and viral origin, camels are exposed to a wide range of internal & external parasitic infestations. Amongst other so many external parasites to which camels are exposed, the Sarcoptic mange is recognized to be one of the most Introduction 3 serious and damaging disease. This disease is caused by a mite known as Sarcoptes scabiei var cameli which belongs to genus Camelus of SARCOPTIDAE family in Veterinary Entomology. It is an extremely pruritic, contagious and debilitating skin disease which is very frequently and sudden in onset. It is also ranked as one of the most serious and important disease of the camels. Sarcoptic mange infestation is very common in the areas of thin skin, the head, neck, flanks, medial aspect of thighs or inguinal region, mammary glands and prepuce. The head is usually affected very rapidly as the animal uses its teeth for scratching the affected areas. Besides linking the occurrence of the disease with poor camel management, malnutrition and contact with infected objects, the stray & infected camels also often become a focus of infecting the healthy animals when mingling with them particularly at watering places for drinking purpose (Richard, 1987; Abdel-Rehman et al. 2001). Sarcoptes is a burrowing mite as it penetrates deeply through the skin surface of the infected camel. This burrowing of mites in the skin helps these parasites lead to intense pruritus and exudative dermatitis. In pruritus, mites penetrate deep into muscular areas, damaging the flesh and lowering the quality of meat. The early inflammatory reaction of the host body towards the mites becomes evident in the shape of small popular elevations, invasion and injuries leading to formation of hairless areas, scaly crust formation or scabs on the affected parts and the skin become dark and thickened. Skin of mangy camel show hemorrhages, and subcutaneous odema after the development of fissures in the underlying epidermis (Kumar et al. 1992; Amer et al. 2006). The fertilized female mites create winding burrows or tunnels in the upper layers of the epidermis of the skin of the host animal and feeding on the serous exudate, a liquid oozing from the damaged tissues. The female mites lay about 40-50 fertilized eggs in these tunnels which Introduction 4 hatch in 3-5 days into a six legged larvae. These larvae immediately crawl to the surface and burrow themselves in the superficial layers of the skin and create small molting pockets. In these molting pockets, the larvae molt to next stages of nymph and adult. The adult male then emerges and seeks a female either in the molting pocket or on the surface of skin. After fertilization the female produces new tunnels, either de novo or, by extension, of the molting pockets, lays eggs in these tunnels and a new life cycle starts. The entire life cycle of Sarcoptic mange is completed in 17-21 days. New hosts can be infected through direct transmission by contact between the animals, presumably from larvae, nymph or adult mites, which are commonly present on the skin surface of the infected animal. Indirect transmission of infestation can also take place through the objects or fomites having mange infection, which come into contact with the affected camel, such as harnesses, blankets, baggage tack, tents and tree trunks (Richards, 1987). The pruritus increases as the mites penetrate deeper in the skin (Al-Rawashdeh et al. 2000, Driot et al. 2011, Bekele et al. 2012). Based on the rate of infection camels can be seriously disturbed by the Sarcoptic infestation as they may stop grazing which can lead to a rapid fall in milk production, and deterioration of health condition. With the increase in the irritation due to scabies, the camel rubs, bites and scratches the affected areas in an attempt to reduce the itchiness. Due to rubbing, biting or scratching, the mites move to the periphery affecting the healthy tissues and resultantly affected area spreads. As the disease prolongs, the skin becomes excoriated, leading to hair loss and the development of scabs. These scabs in turn may be rubbed away and a red surface developed. The animal becomes restless due to severe Sarcoptic mange infestation and involvement of most of the body surface. If the diseased animal is not treated in time, the animal loses its health condition, become emaciated and within two, three weeks the acute stage of Introduction 5 disease may give way to more chronic state (Gorakh et al. 2000, Abubakar et al. 2002, Driot et al. 2011). Sarcoptic mites rarely survive long off the host under natural conditions. A continuous direct contact of animal keepers with their camels can also lead to transmission of diseased condition in human beings which is termed as pseudo scabies. Transmission of infection from camel to man usually takes place during milking, handling or riding. The main symptoms of pseudo scabies can therefore be seen in the inter digital spaces of the hands, on the wrists, forearms, the elbows, the axillary folds and inner side of the thighs. Once a herd is infected with Sarcoptic mange, continuous reinfection of the disease occurs (Schillinger 1987, Singh & Veer 2005, Premalatha et al. 2010). Sarcoptic mange is usually considered to be a seasonal disease and is often reported severe during the winter months as in cold weather the disease had an acute course. However, there is some evidence that in some countries hot weather predisposes to acute outbreaks of camel mange and in the cooler, winter season the rate of mange infestations are at the lowest. In the summer the activity of the mite seems to decline or disease becomes chronic. Dietary intake is an important factor in mange infestation. Nomadic camels on a low nutrition plan, probably carrying heavy worm burdens in hot desert conditions are likely, therefore, to be highly prone to Sarcoptes at this time (Dinka et al, 2010). During such periods of great activity, the mites are readily transmissible from one animal to other animals (Richards, 1987, Banaja & Ghandour, 1994, Tefera & Gebreah, 2001). Mange can easily be diagnosed clinically from the occurrence of pruritus, depilation, alopecia, thickened skin, folds around the joints and encrusted plaques being the main characteristics of this parasitosis. In order to control this zoonotic disease, it is essential to treat Introduction 6 both camel and man along with effective checks over other predisposing factors of the disease such as hygiene and nutritional requirements of the animals. The skin diseases like the scabies both in human beings and animals are being treated with a variety of allopathic drugs now a day, but the role of herbal plants in use since centuries in different shapes cannot be ignored at all, especially in the rural lifestyle. Further with the continuous use of different acaricidal drugs, the issue of resistance development has come across as a challenge for the researchers to find some alternatives for the purpose. Accordingly the research work on the use of traditional herbal medicines is gaining attention day by day. Although there are many reports and studies regarding the prevalence of Sarcoptic mange in camel from different parts of the world, only few preliminary reports are available for Pakistan and none of them provide detailed epidemiology of Sarcoptic mange and its effect on host health. Therefore, keeping in view the importance of the mange problem in camel population of the country, the present project was designed to determine the prevalence of Sarcoptic mange infestation, factors in its occurrence its zoonotic importance, effect on blood physiology and different treatment options in the camel population of Punjab, province in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 2190,T] (1).

4. Isolation and Identification of Parasites From Available Raw Vegetables

by Sawera Chaudhry (2008-VA-189) | Prof. Dr. Azhar Maqbool | Dr.Sarwat Naz | Dr. Muhammad Latif | Dr. Muhammad Avais.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Blank CD. Availability: Items available for loan: UVAS Library [Call number: 2232-T] (1).

5. Taxonomy And Control Of Flea Infestation In Cats At Lahore

by Umair Tariq (2008-VA-233) | Dr. Nisar Ahmad | Prof. Dr. Azhar Maqbool | Dr. Syed Saleem Ahmad.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: INTRODUCTION Fleas play an important role in causing clinical skin disorders and diseases transmission in man and pets animals (Rust & Dryden, 1997). Fleas are one of the most important ectoparasites with more than 2,000 species worldwide affecting mammals, birds, and reptiles (Hsu, 2003). In some locations, fleas represent over 50% of all the dermatological cases presented to small animal clinics. Most are limited to hosts with nests as this can provide conditions for the completion of their life cycle (Linardi & de Avelar, 2014). While fleas on pets are generally considered a nuisance that may cause some dermatologic problems, they are also responsible for the transmission of several important diseases in humans and animals (Dryden & Rust, 1994). They have been involved in transmission of cat scratch disease (Bartonella henselae) (Chomel et al., 2006; Comer et al., 2001), Rickettsia typhi (Murine thyphus), Rickettsia felis (Finkelstein et al., 2002; Rolain et al., 2005), and also serve as the intermediate host for the tapeworm Dipylidium caninum (Rust & Dryden, 1997) and several trypanosomatids (Coutinho & Linardi, 2007). The term ‘‘cat flea,’’ which is the approved common name for Ctenocephalides felis felis (C. f felis), can occasionally cause confusion. When it appears in print, it refers to the specific flea genus and species and not to fleas recovered from cats. There are four recognized subspecies of C. felis throughout the world: Ctenocephalides felis damarensis and C. felis strongylus occur primarily in East Africa, C felis orientis occurs in India and Australia, and the widespread C. f felis occurs in all continents except Antarctica and is the only subspecies that occurs in North America (Rust & Dryden, 1997). The cat flea, C. felis, is a clinically important parasite of domestic pets, being responsible for the production of allergic dermatitis, serving as the vector of Introduction 2 various bacterial pathogens, and being the intermediate host for filarid and cestode parasites. Flea allergy dermatitis is the most common dermatologic disease of dogs and a major cause of feline miliary dermatitis (Dryden & Rust, 1994; Rust & Dryden, 1997). Clinical features vary from asymptomatic to severe hypersensitivity reactions with restlessness, alopecia from scratching and biting resulting in a pruritic papular dermatitis. Vacuuming of carpets, furniture cushions, rugs, or other substrata, with a vacuum machine containing a ‘‘beater bar,’’ will remove many of the flea eggs and larvae. In addition, cocooned pupae at the upper levels of the carpet can also be affected. The vibration also stimulates adult fleas to emerge from their cocoons so that they can be collected in the vacuum machine. Therefore frequent vacuuming, during a flea infestation, can reduce the overall flea burden in the home. It should be ensured that vacuum bags are disposed of properly, to prevent recolonization of the home with flea stages previously removed by vacuuming. Because outdoor development of immature flea life stages is limited to shaded areas, altering outdoor environments to eliminate such habitats can effectively reduce flea populations. Because urban wildlife, such as opossums, raccoons, and foxes, are good hosts for cat fleas, pet owners should avoid encouraging visitations by wildlife, which will affect flea and tick control (see later discussion). Treatment of indoor and outdoor environments with insecticides requires knowledge of what to use and where to use it. For this reason, it is suggested that pet owners consult with a licensed pest control specialist for such applications (Angelbeck-Schulze et al., 2014; Perrins & Hendricks, 2007). In line with increasing urbanization over the last few decades, flea species that infest pets have become household pests. Thus, and for reasons of animal and human welfare, the control of fleas is of great importance worldwide. Despite the increase in the number of products available and Introduction 3 their use, flea infestation of cats and dogs is still widespread in Europe and on other continents, whereas resistance of these insects against many chemicals has been detected (El-Gazzar et al., 1986). Cat fleas are the most important ectoparasite of cats and dogs worldwide. During the past ten years, topical and oral applications of insecticides such as fipronil, imidacloprid, lufenuron and, most recently, selamectin have revolutionized cat-flea control. Recent studies show that these therapies eliminate the need to treat indoor and outdoor environments, and their use markedly reduces the severity and prevalence of flea allergic dermatitis. Surveys have yet to reveal the development of insecticide resistance to these chemical compounds. Extending the longevity of these effective host-targeted therapies should be a major goal of the veterinary community (Rust, 2005). Availability: Items available for loan: UVAS Library [Call number: 2253-T] (1).

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