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1. Experimental Treatment Of Bovine Brucellosis Using Phyto-Chemo-Immuno Theraputic Agents

by Muhammad Kaleem (2013-VA-860) | Prof. Dr. Aneela Zameer Durrani | Dr. Ihtisham Khan | Dr. Muhammad Hassan Saleem | Prof. Dr. Khushi Muhammad.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Bovine brucellosis is very important zoonotic disease in respect of outbreak that resulted in reproductive loses through storm of abortions in herds and infertility in bulls. Currently, no phyto treatment for complete recovery from brucellosis is available. Phyto Chemo-immune therapeutic agents are effective for recovery from bovine brucellosis (Hypothesis) Group A was comprise five adult cows (n=5) positive for brucellosis. Animals in this group were given 3 shots of Oxytetracycline LA at 20mg/kg BW (IM) repeated every 48 hrs. Streptomycin @ 13mg/kg BW IM was given for six consecutive days. Furthermore, flunixin meglumine @ 2mg/kg BW IM for 3 days in combination with antibiotic. Selevit injection containing selenium and vitamin E was administered (IM) for consecutive five days. At next day all the animals in this group were vaccinated with a combined vaccine of RB51 and FMD (Oil based) at a dose of 3mL/animal through deep IM route. In group B, all the animals were given Saafi (Herbal Product) orally consecutively for six days. Furthermore, flunixin meglumin @ 2 mg/kg BW IM was administered for three days. All these treatments were given simultaneously. Then injection Selevit containing selenium and vitamin E were administered (IM) for consecutive five days. After treatment all animals were vaccinated with RB51 + FMD (oil based) 3mL/ animals deep intramuscularly. In group C, flunixin meglumin @ 2 mg/kg BW IM was administered for three days. After NSAID, Selevit injection containing vit. E and selenium was administered (IM) for five days regularly. After treatment, animals were vaccinated with RB51 + FMD (oil based) 3 mL/ animals (IM). In D group five negative adult animals were included in this group and vaccinated subcutaneously with RB51 and FMD (Oil based) at a dose of 3 mL/animal. Summary 40 Data regarding this study was analyzed with ANOVA and completely randomized design was used to compare the mean between different groups using statistical package for social science (SPSS) version 20. P < 0.05 was considered significant. The results showed OD values after 1st, 3rd and 6th month were significantly different in all groups. The mean values depicted that OD values was significantly higher (2.886±0.099) in vaccinated (positive) animal’s vs the other three groups. There was no significant difference in OD values of herbal treated, antibiotic treated and health control animals. The lowest OD values was found in healthy control animals, though significantly similar with herbal treated, antibiotic treated. The mean OD value at 3rd month after treatment is significantly higher (1.794±0.090) in herbal treated and vaccinated positive groups but the OD value of healthy group was lowest (1.794±0.090) after 3rd month of treatment. After 6th month of treatment the OD value of positive vaccinated group was found significantly highest (1.146±0.194). On the other hand lowest value (0.595±0.079) was found in healthy control group. All the groups have found non-significant difference in all group. Regarding Real time PCR the value range from 0-40 Ct values. In the present study the Ct values of the control positive was found to be 13 which indicate control positive. The antibiotic treated group had a Ct value found in this range 35.61 to 38.13. The herbal treated group were all positive and the values were 31.34 to 37.42 Ct. The vaccinated immune booster group. The Ct values in the group was found to be 30.83 to 34.21. The birth weight and placenta dropping time was normal in antibiotic treated animals. There were no significant results found in all groups. Summary 41 This study concluded that herbal regimen is effective in vitro against brucellosis but in vivo it is yet not to be evaluated. The antibiotics can be effective to treat the Brucellosis with this protocol. This was help to control bovine brucellosis, and extensive economic losses. Comparative efficacy of four commonly-immune therapeutic agents were help in choosing the most effective therapy/method for the recovery of bovine brucellosis. Availability: Items available for loan: UVAS Library [Call number: 2388-T] (1).

2. Prevalence Of Fasciolosis In Sheep And Goats Under Range Management Conditions In Azad Jammu And Kashmir

by Imtiaz Ahmad (2009-VA-535) | Prof. Dr. Aneela Zameer Durrani | Prof. Dr. Muhammad S. Anjum | Prof. Dr. Kamran Ashraf.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Fasciolosis is the disease of sheep, goats, cattle and other ruminants. Human and equines are unusual hosts in which instead of liver the flukes may found in lungs or under the skin. Transmission depends on an intermediate host lymnae snail. Animal ingest metacercaria, the worm migrates to the liver where it causes extensive damage and mature worm lives in bile duct. The disease occurs as an acute, sub-acute or chronic infection. Chronic Fasciolosis characterized by anemia, hypoalbuminaemia, emaciation, submandibular edema and loss of condition. Clinical disease is well known but sub clinical infections are often unnoticed, leading to marked economic losses, reduced milk yield, weight loss, reduced fertility and immunity, consequently leading to significant economic losses. Fasciolosis has recently been recognized as an emerging zoonotic disease. Infections in human may be asymptomatic but sometimes nonspecific pain in abdomen, anorexia, dyspepsia and vomiting may occur. Pain in right hypochondrium, epigastrium and jaundice occurs in chronic phase. Sometimes ectopic migration of worm causes abscesses in many organs. The present study investigates the prevalence of Fasciolosis in sheep and goats at different geographic locations on the basis of altitudes in Azad Jammu and Kashmir. 4662 (sheep n=2242; goat n=2420) fresh fecal samples from sheep and goats were collected from three village/towns each of Mirpur, Poonch and Muzaffarabad Divisions. Stool samples were collected from 566 pastoral families of AJK to determine the zoonotic potential of the disease. Prevalence of the disease was calculated on the basis of centrifugal floatation and sedimentation techniques. The intensity of infection was calculated using the McMaster egg counting technique. The risk factors of the disease studied included altitude, season, sex, age, and effect of deworming and flock size on the rate of prevalence. An overall prevalence was recorded as Summary 111 17.88%. Prevalence of Fasciolosis in sheep revealed 26.49% and that of goats 9.91%. The data was analyzed using Chi-square test which revealed a significant difference (P<0.05) in the prevalence of the disease in sheep and goats. The overall prevalence rate in both species was recorded as15.09% at altitude <3000 feet, 25.00% at 3000-6000 and 15.74% at >6000. The highest prevalence was recorded at an altitude 3000-6000 feet. Chi- square values showed significant difference (P<0.05) among three different altitudes. The altitude of 3000-6000ft showed a significantly higher (P<0.05) prevalence of Fasciolosis in sheep and goats. The overall prevalence showed 13.93% rates in spring and 21.77% in autumn. Chi-square values showed a significant difference (P<0.05) in the prevalence of the disease, higher in autumn than Spring. Sex wise prevalence showed 16.67% in male and 18.59% in female animals. The data showed no significant difference (P>0.05) in Chi-square analysis. The prevalence of the disease in the age group below 1 year was 04.40%, 1-4 years revealed 17.73% disease and 36.18% in >4 year. The data showed significantly different (P<0.05) rates in all age groups. Highest prevalence was recorded in sheep and goats above 4 year of age and lowest in those below 1 year. The prevalence in animals with no recent history of deworming was recorded 23.22%. The data showed 15.37% disease in small flocks of sheep and goats <30 as compared to 18.72% in large flocks >30. Chi-square showed a significantly higher (P<0.05) prevalence of the disease in large flocks. Generalized Linear Model (GLM) was used to evaluate the contribution of risk factors (epidemiological factors) to the variations in the prevalence of Fasciolosis in sheep and goats. All the epidemiological factors i.e. altitude, species, season, gender, age group, deworming and flock size were processed. The deworming appeared to be the most significant factor in the model contributing maximum variations in disease with highest Odds followed by age groups, Summary 112 species, season, altitude, flock size and gender. The risk factors for the Fasciolosis in sheep and goats were found, lack of practice of deworming, age group >4 year, species sheep, season Autumn, altitude 3000-6000 and flock size >30. The 75% of the disease prevalence was due to above mentioned risk factors. The deworming, specie goat, age group <1 year, season Spring, altitude <3000, and flock size less than 30 were appeared to be the protective factors in the Generalized Linear Model. The intensity of infection was analyzed through Factorial analysis for difference in species, altitude and season. The difference in eggs per gram of feces was found significantly different (P<0.05) in sheep and goats. The effect of season on egg per gram (EPG) of feces showed a higher mean values in sheep (191.49) and goats (219.72) in Autumn as compared to 158.04 and 180.61 in Spring. In both seasons the mean for goats was found higher than sheep. The effect was found significant (P<0.05), higher during Autumn. Factorial analysis of the data showed significant interaction (P<0.05) between species and altitude. The data showed mean values for sheep 174.04, 191.87 and 168.33 at altitude <3000, 3000-6000 and >6000 feet respectively. The mean values for goats were 232.22, 194.95 and 170.59. The data revealed higher mean for goats as compared to sheep on all three altitudes. Goats revealed significantly higher (P<0.05) number of EPG. POST HOC Tukeys test showed a non-significant difference in intensity of disease between <3000 and 3000-6000 feet, rest of the differences were significant (P<0.05). The overall prevalence in pastoral communities of AJK was 0.88%. The samples were collected from male and female of 4 age groups <10 year, 11-20, 21-40 and >40. The prevalence in male was 0.76 and in female was 0.98%. The data showed that age groups below 20 year were Summary 113 the susceptible groups in both sexes. The highest prevalence (2.25%) was found in female age group 11-20 year. Age groups above 20 year did not revealed any positive sample. 227 adult liver flukes were collected from livers of infected animals of different animal species (sheep, goats. cattle and buffaloes) and geographic locations for morphometric and molecular identification of the species of Fasciola. Flukes were identified on the basis of measurements of body length, body width, diameter of suckers, distance between oral and ventral sucker and distance between ventral sucker and posterior end of the body. The measurements of F. hepatica showed a body length range 13-34mm with an average length of 21.51mm whereas, F. gigantica ranged from 28-52mm with an average of 42.27mm. The average body lengths of F. hepatica below 3000ft was 21.9, at 3000-6000ft was 21.07 and above 6000 ft was 22.00mm and that of F. gigantica was 42.05 and 42.44mm at 3000 and 3000-6000 feet. The measurements of F. hepatica revealed an average body width of 10.05mm, average diameter of oral and ventral suckers of 0.74 and 1.28mm respectively and average distance between the two suckers of 1.34mm. The readings for F. gigantica were 9.46, 0.89, 1.55 and 1.72mm respectively. Average distance between ventral sucker and posterior end of the fluke in case of F. hepatica was 18.35mm at all three altitudes and host species of animals and it was 38.26mm in case of F. gigantica. The overall mean worm load was 13.56 worms per liver of animal with a range 5-26. The mean worm load of F. hepatica was 10.9 and that of F. gigantica 13.11. Mixed infections were noted at altitudes below 6000 ft. Infestation with F. gigantica was not found at altitudes above 6000ft and F. hepatica was encountered at all three altitudes. 51.98% of the recovered flukes were F. gigantica which showed the equal chances of infection with either species of Fasciola in AJK. The results revealed that F. hepatica was the fluke affecting animal population at altitude above 6000 ft and F. gigantica was the major fluke below Summary 114 3000 ft. At altitudes between 3000-6000ft, 36.20% of flukes were F. hepatica while its prevalence was reduced to 17.30% below 3000 ft. Once the species of the liver flukes were identified morphologically they were subjected to molecular conformation through amplification of the genomic DNA of the two species through PCR using two sets of species specific primers. In the PCR based on primer set 1, a product of 391 bp was generated from the genomic DNA of Fasciola hepatica whereas no product was generated from the DNA of Fasciola gigantica. PCR based on primer set 2 amplified a 235-bp product from the DNA of Fasciola gigantica. The molecular identification in the present study showed that morphometric identification of the two species is valid and standard population of both species were found present at different geographic locations and species of the animals of the state of Azad Jammu and Kashmir except F. gigantica not found above 6000 feet altitude. The ethno veterinary practices for Fasciolosis were documented through Participatory Rural Appraisal. A total of 173 respondents/key informants were interviewed during the study period in the study area. The majority of the traditional healers (n=33) elders of pastoral families (n=53) and sheep/goats owners (n=56) were above the age of 40 year. Veterinary officers (n=6) and assistants (n=25) were interviewed as a part of verification process. 31.69% of the respondents were found using allopathic anthelmintic along with ethno veterinary medicines. 53.52% of the respondents were using ethno veterinary medicines because of non-availability or cost effectiveness of allopathic anthelmintic. 95.18% of the respondents were using plants or part of the plant as traditional anthelmintic in their sheep or goats.18 plant families were identified during the survey which include Acanthaceae, Asclepiadaceae, Asteraceae, Berberidaceae, Boraginaceae, Cannabinaceae, Chenopodaceae, Euphorbiaceae, Fabaceae, Summary 115 Gentianaceae, Juglandaceae, Liliaceae, Malvaceae, Oxalidaceae, Punicaceae, Rhamnaceae, Scrophulariaceae, Solanaceae. The species of medicinal plants identified during the study were Berberis lyceum, Nicotiana tabacum, Asparagus officinale, Calotropis procera, Aloe vera, Mallotus philippensis, Adhato davesica, Artemisia scoparia, Xanthium strumarium, Chenopodium ambrosoides, Artimisia maritime, Verbascum Thapsus, Acacia Arabica, Cordlia myxa, Cannabis sativa, Rhamnus purpurea, Juglansregia, Oxalis corniculata, Punica granantum, Artimisa fragrans, Swertia petiolata and Abutilon indicum. Availability: Items available for loan: UVAS Library [Call number: 2430-T] (1).

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