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Molecular Epidemiological and Pathological Studies on Paratuberculosis in Cattle and Buffaloes

Thesis Info

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Author

Aziz Ur Rehman

Program

PhD

Institute

University of Agriculture

City

Faisalabad

Province

Punjab

Country

Pakistan

Thesis Completing Year

2017

Thesis Completion Status

Completed

Subject

Pathology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/11303/1/Thesis%20final%20after%20HEC%20%282005-ag-1052%29.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726699154

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Bovine Paratuberculosis is a chronic disease primarily of the gastrointestinal tract. The disease is caused by Mycobacterium avium subsp. paratuberculosis (MAP). There was limited information available about this disease in the country and there was need to have an understanding about the magnitude and pathology of disease under local conditions. The study was planned to measure the Paratuberculosis prevalence in cattle and buffaloes at twelve government livestock farms , two cattle/buffalo colonies and the slaughterhouse of Faisalabad. A total of 2181 animals from twelve livestock farms, 265 cattle and buffaloes from two cattle/buffalo colonies and 200 consecutive cattle and 200 consecutive buffaloes from the slaughterhouse of Faisalabad, were included in this study. Tuberculin testing was performed on all the animals (cattle and buffaloes) above two years of age present at farms and colonies. Blood and faecal samples were gathered from tuberculin positive (reactor) animals. These samples were further processed by ELISA, ZN microscopy and PCR. Morbid tissue samples were collected from cattle and buffaloes slaughtered at slaughterhouse for histopathology and isolation of the organism. PCR test was used for further confirmation of Mycobacterium from farms, colonies and slaughterhouse also. The data collected from the study was analyzed by using frequency analysis and logistic analysis procedures. The study showed that the prevalence of Paratuberculosis at government livestock farms was 3.8% on the basis of tuberculin + ELISA test, while in case of two cattle/buffalo colonies it varied from 3.4-14.66%. Multivariate logistic regression analysis in both cattle and buffaloes at twelve livestock revealed that specie, milk production, total animals, total small ruminants and total buffaloes showed significant association with occurrence of Paratuberculosis while, the bivariate logistic regression analysis in cattle and buffaloes revealed that farm number, age and number of the total cattle showed significant association with occurrence of Paratuberculosis. Multivariate logistic regression analysis revealed that herd, age and lactation number showed significant association with the occurrence of Paratuberculosis, while the bivariate logistic regression analysis revealed that herd showed significant association with occurrence of Paratuberculosis. The slaughter house of Faisalabad was also included in the study. Out of total 200cattle and 200 buffaloes, suspected morbid samples of intestine and mesenteric lymph nodes from 25 cattle and 20 buffaloes were collected. These samples were further processed for histopathology and PCR. The main microscopic lesions observed in intestine were the diffuse inflammatory reaction with mononuclear cell infiltration and degenerative and necrotic changes in sub-mucosal glands along with the formation of immature granulomas. Relationship between Zn and PCR showed that 50% animals were found positive by both ZN and PCR. To compare the results of ELISA and PPD, an experiment was carried out on total 140 adult animals at Livestock Experimental Station University of Agriculture, Faisalabad. This revealed that ELISA showed more positive results than tuberculin testing. The study concluded that that tuberculin testing, ELISA, ZN staining and PCR are efficient diagnostic tools to diagnose the disease and use of combination of different tests improves the efficiency and confirmation of the disease.
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منشی محمد احتشام علی کاکوروی

منشی محمد احتشام علی کاکوروی 
لکھنؤ کی سرزمین میں اپریل کے چوتھے ہفتہ میں ایک اور حادثہ پیش آیا یعنی کاکوری کے ممتاز خاندان کے رئیس جناب منشی محمد احتشام علی صاحب نے ۲۲؍ اپریل کی صبح کو ۷۵ برس کی عمر میں وفات پائی، کہنا چاہئے کہ اودھ میں قدیم شریفانہ جوہر وضعداری، دینداری، مروت، سیرچشمی، غربا نوازی اور مسکین پروری کا یہ اخیر نمونہ تھا، ان کی پوری زندگی میں جس میں وسعت کا زمانہ بھی تھا اور تنگی کا بھی، ان کے ہاتھ یکساں کھلے رہے اور اس اخفاء کے ساتھ کہ بائیں ہاتھ کو بھی داہنے ہاتھ کی خبر نہ تھی، وہ مولانا شاہ فضل رحمان صاحب گنج مراد آبادی سے بیعت تھے، اس تعلق کو اخیر اخیر وقت تک جس طرح نباہا، وہ ان کی سعادت مندی کا نشان ہے، پابندی یہ کہ مرتے وقت سجدہ عبودیت ادا کیا ہے اور صبر و شکر کے کلمے زبان سے نکلتے رہے۔
ان کی جوانی تھی کہ ندوۃ العلماء کا غلغلہ بلند ہوا، چونکہ اس مجلس کے سرپر فضل رحمانی سایہ فگن تھا۔ اس لئے حضرت شیخ کے سارے حلقہ بگوش اس کے حلقہ میں تھے اور اسی مناسبت سے جناب منشی محمد اطہر علی صاحب مرحوم اور ان کے ساتھ میں جناب منشی احتشام علی صاحب ندوہ کے خدام میں داخل ہوئے تھے، اپریل ۱۸۹۵؁ء میں اس کا پہلا جلسہ لکھنؤ میں ہوا تھا، اس اپریل ۱۸۹۵؁ء سے لے کر ۲۲؍ اپریل ۱۹۴۳؁ء کی صبح تک جب کہ انہوں نے زندگی کی اخیر سانس لی ہے، یکساں دلچسپی خلوص و انہماک سے اپنے فرائض کو انجام دیا ہے۔ نہ صرف رئیسوں میں بلکہ مسلمانوں میں اس قدامت خدمت اور مخلصانہ مذہبی خدمت گزاری کی مثال شائد ہی ملے۔
خیالی گنج میں ان کی بڑی اور وسیع کوٹھی، ان کے عزیزوں کا...

The Perception of Manobo Tribe on Different Aspects of Life: Their Effects on the Living and Social Conditions in San Luis, Agusan Del Sur

The study attempted to determine the perception of the Manobo tribe and their effect on their living and social conditions in Barangay Sta. Ines, San Luis Agusan del Sur. The descriptive method was used in this study describes the perception among the Manobo tribe on different aspects of life: their living and social conditions.  This study was conducted in barangay Sta. Ines, san Luis Agusan Del Sur.  There were only 195 of them who were taken as the respondents and researcher used the standardized questionnaire which were taken from the book of Mendez and used by the previous researchers. Hence, no validation was needed.  The random sampling design was used in this study in selecting the respondents. On the level of perception among the Manobo tribe, the Manobos agree on their belief in god as supported with the total average weighted mean ranging from 2.01 to 2.73 or agree.  They strongly agree to get involved in social gathering as evidenced with total weighted mean of 2.21 or agree.  They strongly agree on good aspiration as evidenced with average weighted mean of 2.71 or agree.  They agree on the family practices as supported with the average weighted mean of 2.45 or agree.  They strongly agree on good family upbringing accompanied with an average weighted mean of 2.57 or strongly agree.  They also strongly agree on how to treat visitors accompanied with an average weighted mean of 2.79 or strongly agree. On the living conditions, the Manobos rank 1 that they do not depend their main source of income but they work hard to have additional income for their family needs.  They also rank 1 that their house is made up of wood, Nipa and bamboos for the shelter of their family and the Manobo rank 1 that they usually used herbal for their natural medicine but they send the sick person to the hospital for the health of the family. On the social conditions, the Manobos strongly agree to establish a neighborhood relationship with an average weighted mean of 2.78 or strongly agree.  They also agree to establish kinship relationship and community relationship as indicated with an average weighted mean of 2.84 and 2.78 or strongly agree respectively. On the effect of perception on different aspects of life, belief in God, social; gathering, family practices, family upbringing and aspiration are best predictors in the financial condition of Manobo.  Aspiration, family practices, treating visitors, wedding practices and social gathering are best predictors in neighborhood relationship.  Wedding practices, treating visitors, aspiration, family practices, social gathering, family upbringing and belief in God as the best predictors in kinship relationship Family upbringing, aspiration, and family practices are best predictors in community relationship.

Designing, Construction and Evaluation of Effective Dual Digestion System for the Treatment of Wastewater and Sludge

Advancement and further standardization of contemporary municipal wastewater and sludge treatment facilities is underway in order to increase their efficiency and promote environmentally safe sustainable practices. Practically, a dual stage aerobic/anaerobic–anaerobic system has always been a more viable approach in tackling wastewater of domestic origin. Typically, the amount and the quality of sludge generation and the effective temperature management keeping in view the cost-effectiveness and environmental health during treatment of domestic wastewater and sludge are vital. A more comprehensive approach to deal with wastewater and its sludge along with production of biogas and Class A biosolids is desired. Therefore, the study is aimed at improving the efficiency of the dual digestion system (sequential aerobic/anaerobic-anaerobic) for wastewater and sludge, producing Class A biosolids coupled with energy (biogas) generation under optimum environmental conditions. Besides, a detailed investigation of culture and molecular based bacterial diversity and density was also carried to highlight the involvement of specific bacteria. In the 1st phase, laboratory scale sequential aerobic and anaerobic system was developed for the treatment of domestic wastewater under the influence of low to slightly high temperature regime (5, 25 and 45°C) and different aerobic retention times (1-3 days). Overall, the treatment efficiency varied from 92-100 % in terms of biochemical oxygen demand (BOD), Chemical oxygen demand (COD), turbidity and nitrite nitrogen (NO2-N) removal in total retention time of 14 days (2 days aerobic and 12 days anaerobic digestion). Increase in aerobic retention time from 1-3 days improved the treatment efficiency by 5-20 %. However, a slight increase though non- significant in COD (94 - 97.1%) and BOD (95.1 - 96.7%) reduction was observed under treatment from 25 to 45°C of temperature. Whereas, nearly 60% decrease in treatment efficiency was observed in terms of BOD and COD removal when temperature reduced from 45-5°C. The low temperature (5°C) treatment efficiency of the whole system was recovered to maximum within 6-8 days when reactor was bioaugmented with activated sludge. A significant decrease (98-99%) in pathogenic bacteria (HPC/mL) (Escherichia coli, Salmonella typhimurium, Shigella dysenteriae, and Pseudomonas aeruginosa) was observed in the remaining biomass (sludge) after treatment. Likewise, the MPN index of fecal coliforms and E. coli in the sludge also showed considerable reduction. Further changes in pollution indicators such as pH and nitrites (2.86–0.01± 0.05 mg/L), nitrates (0.98–0.02 ± 0.05 mg/L), phosphates (0.847–0.50 ± 0.05 mg/L) and sulphates (0.721-0.28 ± 0.03 mg/L) indicated involvement of the other key bacterial species in digestion of wastewater. In the 2nd phase, beneficial nitrifying bacteria were isolated and identified as Nitrososmonas sp. and Nitrobacter sp. from the raw wastewater and activated sludge (feed/influent). Besides, their specific activities were also determined based on different substrates utilization rates. Further beneficial nitrifying, sulphur oxidizing and phosphate accumulating bacteria were successfully screened in the biomass from aerobic phase of sequential aerobic-anaerobic treatment facility at 25°C. In the stable operating conditions, over 98% removal rate for total nitrogen was observed with and without activated sludge seeding. Moreover, COD removal rate reached up to 93%, indicating both organic matter and ammonia removal. Autotrophic bacteria viz. ammonia, nitrite and sulphur oxidizers and phosphate accumulating bacteria were isolated through culture enrichment techniques demonstrated about 49-72% of NH3-N, 60-94% of nitrite, 18-30% sulphur and 25-59% phosphorus removal was observed in activated sludge biomass. These results were also confirmed and correlated with the activities of different hydrolytic enzyme such as Hydroxylamine oxidase, nitrite oxidase, sulphur oxidase and alkaline phosphatase in the activated sludge. In the 3rd phase, the influence of thermal pretreatment (55°C for 2 days) on the mesophilic anaerobic digestion (35 ± 2°C) of secondary and primary-secondary combination of sludge was studied. Besides, the treatment efficiency was also investigated for biogas yield, sludge stabilization and pathogen reduction at 12 and 20 days of solid retention times respectively. The thermal pretreatment (55°C for 2 days) subsequently with mesophilic anaerobic digestion at 35±2°C of secondary and primary/secondary sludge combinations improved the removal of VS by 35-38%, TCOD by 38-43% and SCOD by 37-47% with an overall yield of biogas by 31-34% (0.162-0.174NL/gVSreducecd), methane (0.082- 0.103NL/gVSreduced). However, treatment of secondary sludge showed a slight increase in COD solubilization (1-4%), organic matter reduction (10-11%) and specific biogas yield of 10-20% compared to primary-secondary sludge combinations. Moreover, this sequential mode of treatment helped removal of pathogenic bacteria by 7.6 - 8.1 log10 units thereby meeting the U.S. standard for Class A biosolids. The last phase of the study evaluated the comparative performance of dual stage 45°C thermophilic-mesophilic temperature phased anaerobic digestion system and 55°C thermophilic- mesophilic TPAD system with respect to sludge hydrolysis and methane production under the same operational conditions (6.5% of Total Solid content and 12.5 days total Solid Retention Time). The overall performance of TPAD-I system achieved 77% reduction in volatile solids i.e. only 5% higher than TPAD-II system. There were observed no ammonia inhibition and excessive level of volatile fatty acids accumulation and consequently the two phase digesters were able to yield significantly higher rate of methane production ((45°C, 3.55±0.47 L CH4/L.day; 35°C, 1.44 ± 0.12 L CH4/L.day) than TPAD-II. TPAD-II system suffered from certain degree of instability such as high VFAs accumulation (6087 ± 1578 mg/L), low buffering capacity and increased level of total NH3 (2982 ± 219mg/L) and free NH3 (226 ± 25 mg/L), reduced level of methane production (1.69 ± 0.1 L/L.d) was seen rather than being stopped. The bacterial and archaeal population were investigated using high through-put 454 pyrosequencing and Illumina sequencing respectively. In both TPAD systems, the associated bacterial population was dominated by Firmicutes (45.5-60.9%), Bacteriodetes (20-26.5%), Proteobacteria (8.6- 36%), Synergistetes (2.1-10.6%) and Actiniobacteria (1.9-7.2%), while archaeal community was dominated by Methanomicrobia (Genus Methanosarcina: 74-84%) and Methanobacteria (Genus Methanobacterium: 15-27%). In particular, there was observed a progression from genus Clostridium to Coprothermobacter and Tepidanaerobacter, and Methanocarcina to Methanothermobacter and Methanobacterium in 55°C TPAD. Variation in the composition of microbial populations in the first thermophilic stage at 55°C was attributed to the temperature, that of second mesophilic stage associated bacterial communities was related to the influent coming from first thermophilic stage. This study determined the key bacterial species that were involved with enhanced performance of TPAD systems at different temperature regimes. The overall results proved that dual digestion system has a great potential to be up-scaled at large scale for handling wastewater and sludge for small communities in developing countries even at low temperature conditions. It will not only support to mend the public health in terms of removal of unwanted organic/in-organic compounds and pathogens from wastewater and sludge producing Class A biosolids for large scale safe application in croplands