Etiological Study Of Pancytopenic Children
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Publisher: 2011 Dissertation note: Pancytopenia is a hematological condition in which there is a decrease in all three cell lines of peripheral blood i.e. erythrocytes, leucocytes and platelets leading to anemia, leucopenia and thrombocytopenia. Complications of anemia, repeated infections and bleeding tendencies are sometimes horrifying and may result in death of individual.
The present study was designed to analyze the underlying pathology, different clinico-haematological features and importance of bone marrow study in one hundred children presenting with pancytopenia. Present study was carried out in pediatric laboratory of Mayo Hospital, Lahore, Pakistan..
Detailed history was taken in all cases. Complete blood counts were done on an automated blood analyzer (Sysmex Kx-21). For counter check of total leucocyte count, differential leucocyte count and platelets, smears were also prepared and stained by using Giemsa stain. Red cell morphology was done on blood smear for theconfirmation of red cell indices. A total volume of 3 ml venous blood was drawn into a syringe. Out of which 1.0 ml was delivered into EDTA containing vacutainer and remaining 2 ml blood was transferred to a plain glass tube. After clotting and centrifugation serum was obtained for screening of hepatitis B surface antigen and antibodies against hepatitis C virus. Bone marrow aspiration was also performed where indicated.
Megaloblastic anaemia (42%) Aplastic anaemia (26%) and ALL (8%) were found to be the common causes of pancytopenia in our scenario. Less common causes of pancytopenia were infections (8%), mixed deficiency (4%), MDS (4%) and lymphoma (4%). In all above mentioned cases clinical manifestations and peripheral blood counts played an important role in their evaluation.
Two cases of haemophagocytic syndrome (2%), a rare cause of pancytopenia were also diagnosed in this study.
This study also explained the importance of physical examination, peripheral blood findings and bone marrow examinations in the management of pancytopenic patients. Peripheral blood film and bone marrow aspiration should be performed simultaneously in pancytopenia patients when the diagnosis is not confirmed. Bone marrow examination in most cases gives the specific diagnosis. However, in few cases, additional tests may be required.
Serum vitamin B12 and folic acid levels may be needed for confirmation of megaloblastic anemia. Serum iron, TIBC and iron staining on bone marrow smears may be required in iron deficiency anemia. In cases of leukemia flow cytometry study may be more helpful in reaching a final diagnosis. Bone marrow biopsy is mandatory in aplastic anemia.
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Clinicopathological Study Of Theileriosis In Naturally Infected Sheep
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Publisher: 2013 Dissertation note: Theleiriosis in sheep is an important infectious disease of small ruminants characterized by General weakness, weight loss, anorexia, elevated body temperature, petechial hemorrhages on conjunctival mucosa, swollen lymph nodes, anemia and cough. Changes in hematological, serum biochemical and histopathological parameters are good indicators for diagnosis of theileriosis. For this purpose a total of 100 blood samples from infected sheep as well as 50 blood samples from healthy sheep were collected on the basis of clinical examination and peripheral blood smears from different farms in and around Lahore. The collected samples were processed at Clinical Pathology Laboratory, Department of Pathology, University of Veterinary & Animal Sciences, Lahore. Hematological parameters were measured by automatic hematology analyzer & serum biochemical parameters were measured by using commercial kits by colorimetric methods using spectrophotometer. Histopathological changes were observed by making slides of tissue samples, and observed under microscope. Molecular confirmation of Theleria sp. was done by using Polymerase Chain Reaction. Finally the data obtained for hematological and serum biochemical parameters were analyzed by using Student's t test.
Previous studies on serum biochemical parameters and hematological parameters were mostly done in large ruminants. But there is little investigation available on the biochemical and hematological alterations of theileriosis in small ruminants. In this study blood samples taken from sheep were examined under microscope by making thin blood smears using Giemsa stain. Theileria piroplasms were seen in RBCs at 100x. Blood samples were also processed for PCR test. In all Theileria ovis positive samples 520-bp fragment was generated on gel by using primers TSsr 170F and TSsr 670R for sheep. All samples positive on microscopy were also positive by PCR. This study provides the base line data for molecular diagnosis of Theileria ovis in sheep in Pakistan. Hematological and biochemical parameters were evaluated in the sheep naturally infected with theileriosis while healthy sheep were selected as control. Hematological findings revealed a significant decrease (P<0.05) in RBC count, WBC count, Hb Conc. and PCV in infected animals as compared with healthy animals. Serum biochemical findings also revealed alterations in activities of enzymes and plasma proteins. A significant decrease in total proteins, albumin, glucose and creatinin and triglycerides was observed while significant increase (P<0.05) in ALT, AST, bilirubin was observed in affected animals as compared with healthy animals. A non significant increase in urea and cholesterol concentration was also observed in infected animals as compared to healthy animals. Different tissues were examined to study gross and histopathological changes. Liver was large, pale and friable. Prescapular lymph nodes were swollen and spleenomegaly was also seen. Kidney showed hemorrhagic spots and some were mottled. Histological examination of the lymph nodes revealed edema, widened intercellular spaces, parenchyma degeneration, massive lympholysis, widened sinuses at some areas and macrophages. Splenic nodules were sparse, diminished in size and lymphocyte depletion in follicles was prominent.In liver, mild infiltration of leukocytes around blood vessels, vacuolation in hepatocytes, increase in sinusoidal space and atrophy of hepatic cord was seen and hepatic cord was broken. In kidney, degeneration of tubular epithelium cells was seen. This study concluded that theileriosis in sheep was associated with some alterations in blood parameters and histopathological changes which could be useful in the diagnosis of ovine theileriosis.
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Comparative Pathological Studies Of Brucellosis In Azikheli And Nili Ravi Buffaloes At Slaughter House I District Sawat
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Publisher: 2013 Dissertation note: Abstract
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Cytotoxic Efects Of Saw-Scaled Viper Snake Venom On Brain, Heart, Liver And Hematological Parameters Of Albino
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Publisher: 2014 Dissertation note: Abstract
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Pathobiological Studies On Bovine Ephemeral Fever Infected Cattle In District Swabi
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Publisher: 2015 Dissertation note: Among the non-contagious diseases Bovine ephemeral fever is important disease of cattle the course of the disease is usually three days due to which it is called “three days sickness”. This is transfer to other cattle through insects Culicoides (biting midges a group that include many kinds of flies) and mosquitoes. Bovine ephemeral fever virus (BEFV) has been collected from Culicoides coarctatus, Culicoides brevitarsis, and Anopheles bancroftii (Walker et al. 2012). Cattle and buffaloes are the main species affected from Bovine ephemeral fever (BEF) which gives huge economic losses to the dairy sector. The etiologic agent, Bovine ephemeral fever virus belong to Rhabdoviridae family, enveloped (negative sense) ssRNA virus. It generally recur in Australia, Asia, and Africa also in the Middle East (Walker 2005).
The theme of the present study was detection of BEF virus through Reverse Transcriptase-Polymerase Chain Reaction from the cattle suspected for bovine ephemeral fever virus on the basis of clinical signs. Hematological profile and serum calcium level were checked in the confirmed positive samples for BEFV.
A total of 50 blood samples were collected from the suspected animals in aseptic condition using a sterilized disposable syringe and were preserved in vacutainers (Anticoagulant added n = 50, without anticoagulant n = 50). The 10 blood samples were collected from healthy animals in vacutainers (Anticoagulant added n = 10, without anticoagulant n = 10).
Buffy coat were separated from blood samples and from this the RT-PCR was performed and successfully diagnosed the BEFV infected cattle. Hematology and serum calcium were performed for both confirm positive and healthy animals.
The result showed that BEF virus was diagnosed with the help of RT-PCR in samples suspected for BEFV infection, and there was no virus detected in samples taken from healthy animals. Comparison of hematology between BEFV infected cattle and healthy animals were performed there was no changes in the RBC, Hemoglobin, Hematocrit, MCV, MCH, MCHC, and MID (it include monocyte, eosinophils, and basophils) except Neutrophils, which number was increases and lymphocytes which was decreased in BEFV infection, while in healthy animals there was no change in the whole hematology. Serum calcium was also determined there was decrease in serum calcium level of BEFV infected cattle, while in the healthy animal samples there was no change in the serum calcium level.
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Study On The Pathogenesis Of Co-Infection Of Infectious Bronchitis (Ibv) And Escherichia Coli (E. Coli) In Experimental Chickens
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Publisher: 2015 Dissertation note: Infectious bronchitis and colibacillosis are infectious diseases affecting chicken of all ages and breeds. They are of major economic importance in commercial chicken flocks, causing huge losses. As both in humans and animals it is well documented that preceding respiratory infection of virus predispose individual to bacterial infection. Moreover, mix infection in poultry which occur when different organisms simultaneously invade birds is a major threat to poultry industry causes highly epidemic debilitating disease with high mortality which eventually leads to economic catastrophe. In recent past prevalence studies of field, E. coli had been reported with high prevalence and exaggerated disease along with other respiratory pathogens, additionally IBV had also isolated from same flocks in same season. Although a plethora of pioneering work had been done on IBV and E. coli in the previous decades but still a window in time exist in revealing there co-infection. Looking to field scenario in our country, the present study was designed to study an ideal challenge model for IBV and E. coli, by reproducing the natural infection.
80 SPF day old broiler birds were arranged into four groups, (A, B, C and D). Each group was comprised of 20 birds. Group D served as uninoculated control while, Group A and B were challenged with IBV on 23rd day of trails, and Group B and C were inoculated with E. coli infection on day 26th. Birds, (n=3) from each group were slaughtered on various days post infection, gross and histopathological lesions were observed and serum samples for HI were taken, throughout experiment. Variable clinical signs were recorded in various groups. In IBV infected group, respiratory distress i.e., tracheal rales, coughing, sneezing and gasping were noted during early stages, later up to 10 days post infection watery diarrhea with ruffled feathers were observed. In mix infected group clinical signs manifested rapidly and were persistent with high severity. Gross lesions in mixed infection were more profound,
including; airsacullitis, tracheitis with catarrhal exudation throughout respiratory tract; severe sepicemic lesions i.e. perihepaitis, pericarditis, pneumonia and polyserositis with swollen and pale kidneys distended by urates. 5 birds died in mix infected group revealed ascites with asphyxiation of trachea with caseous exudate. While in IBV infected group lesions were mild and confined to trachea, airsac and kidneys. Mortality was high in mix infected followed by IBV in which two birds died. While in E. coli and control group mortality were not noted. Histopathological lesions in mix infected group were aggravated markedly tracheal epithelium degeneration, deciliation and sloughing; congestion, interstitial nephritis, leukocytes infiltration, tubular degeneration and necrosis while were observed. In lungs, pneumonia of peribronchiolar area and interstitium with lymphocyte and macrophages infiltration, additionally degeneration and vacuolization of hepatocytes with focal necrotic areas were also noted. In IBV and E. coli group microscopic lesions were of mild degree. GMT of both IBV and mix infected birds were high but were not significant different (P>0.05). Among the groups, statistically significant increase in FCR of birds in mix infected group was observed followed by E. coli, with IBV infected came third in the row. On the bases of these findings we might conclude that mix infection of IBV and E. coli causes severe lesions with high morbidity and mortality.
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Pathological Investigation And Molecular Detection Of Avian Pathogenic E.Coli Serogroups In Broiler Birds
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Publisher: 2016 Dissertation note: The present study was designed to identify the serogroups present in field and to study their pathological effects in experimentally infected broiler chicks. The present study was attempted to scan the rfb gene clusters in APEC predominant serotypes O1, O2 and O78 strains and to develop Multiplex PCR method for serotyping of the O-antigens. The Multiplex PCR method was used for the identification of serotypes of APEC. The second part of the study was to study the pathological lesions caused by most prevalent serogroup in experimentally infected broiler chicks.
A total of 100 tissue samples (lungs and livers) were collected from colibacillosis suspected broiler birds. Streaking was done from these samples on three different media and it was found that 80% isolates were positive on MacConkey media, 60% were positive on EMB media and 40% were found pathogenic for E.coli on Congo red media.
The colonies which were of pink color on congo red media were considered as pathogenic. DNA was extracted from these colonies by boiling method by picking single colony from each petri plate. Extracted DNA was further used for PCR to confirm the three serogroups i.e O1, O2 and O78.
The PCR results showed that 8% isolated samples were found as pathogenic as O2 strain was found dominant among all. Only two genomic DNA samples were found of O1 serogroup After confirmation of serogroups inoculum of Avian pathogenic E.coli O2 strain was prepared to experimentally infect the broiler birds. Birds were infected at the age of day 7 via intratracheal route.
Following the experimental infection of birds, they were monitored for any pathological lesions which were not present significantly while some birds were off feed, reluctant to move, head down posture and were keeping themselves in isolation.
Postmortem of dead birds was performed and pathological lesions were noted. Livers were found to be congested, enlarged and white fibrinous layer over liver was present. Lungs were also affected with the disease and white layer was present on lungs too. Lungs were consolidated and congested.
Histopathology of lungs and livers was performed. It was noted that there was mononuclear cells infiltration and thin fibrinous layer over liver. Thickening of the liver capsule was noted due to infiltration of mononuclear cells and there was marked congestion in hepatic portal areas and the central vein. There was atrophy of adjoining hepatic cords due to greatly distended and congested sinusoids. Besides these changes, hepatocytes in various stages of degeneration along with hemorrhages, areas of congestion and fatty changes in a few places could be seen.
There was infiltration of heterophils, severe congestion, lymphocytes and macrophages in the wall of the bronchus as well as in the peribronchial alveoli. There was marked presence of granuloma in lungs. Some birds displayed thickening of the pleura and consolidated areas covered with yellowish fibrin in lungs.
The experimental infection of avian pathogenic E.coli confirmed the hypothesis that it causes pronounced histopathological lesions in broiler birds.
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A Study On Point Prevalence, Etiological And Biochemical Investigations Of Post Parturient Haemoglobinuria In Buffaloes In Tehsil Bhalwal
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Publisher: 2017 Dissertation note: Post-parturient haemoglobinuria is a disease of great economic importance of sub-continent affecting a large number of buffaloes. It is characterized by intravascular hemolysis, haemoglobinemia, haemoglobinuria ultimately leading to anemia. The exact pathogenesis is yet unknown as there are many diversified etiological factors have been associated with this disease. All the relevant information is relatively scanty. Consequently present study has been aimed to study all possible risk factors associated with this disease in tehsil Bhalwal of district Sargodha where a large number of increasing cases were reported by the local governmental body. Etiological, hematological and biochemical risk factors were quantified to facilitate control measures and upcoming research priorities.
This study was conducted from the period of about 4 months from November 2016 to February 2017. Cross-sectional epidemiological observations were documented on hemoglobinuric and healthy buffaloes for hematological and biochemical study related to parturient haemoglobinuria. The sample size was determined to three hundred and eighty four animals.Present study was observed during the period of four months (November 2016 to February 2017). Out of 384, forty animals (n=40) were confirmed with post parturient hemoglobinuria. The point prevalence observed during the period of four months was 10.4%.
Buffaloes showing signs of hemoglobinuria along with parturition history, pale mucous membranes, mild tachycardia and dyspnoea was assumed as affected with post-parturient haemoglobinuria while animals suffering from other problems like babesiosis causing red urine were omitted from the study after verification of diagnosis through giemsa staining. The blood samples were processed for haematological analysis for the final confirmation of positive
haemoglobinuric buffaloes. Blood sample collected and placed in EDTA vacutainerswas processed for hematology to study hemoglobin (Hgb) values, total erythrocytes count (TEC), erythrocytes sedimentation rate (ESR) and hematocrit (Hct), total leukocyte count (TLC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) in addition to mean corpuscular hemoglobin concentration (MCHC) by using haematological analyzer. Haematological analysis of all the samples was made from Department of Pathology, UVAS, Lahore.Serum samples of all buffaloes were analyzed for biochemical analysis asalkaline phosphatase (ALP), serum urea, glucose,bilirubin, creatinine, calcium, phosphorus, copper, and molybdenum. Moreover, urinalysis was done for gross and biochemical analysis.
Results of the study revealed significant difference among complete blood count (CBC) includingHgb, TEC, Hct and TLC, ESR, MCV and MCH. However, there was no significant variation among MCHC values in affected buffaloes. Serum biochemistry also revealed significant difference of various parameters including ALP, creatinine,BUN, total bilirubin, phosphorus, copper and molybdenum. However, no significant difference was detected among the healthy and affected groups regarding blood glucose and serum calcium levels. There was significant elevation in pulse and respiration rates in buffaloes affected with hemoglobinuria.
The results regarding mineral analysis of the soil shows significant difference in phosphorus and copper. Moreover, mineral levels of soil and serum of animals showed significant relation of phosphorus levels, followed by the levels of molybdenum. Calcium and copper levels also showed moderate relationship.
Observations regarding parity/lactation number reveal the highest incidence rate of 35% among buffaloes at 3rd lactation, followed by buffaloes at 4th, 2nd, 5th, 1st and 6th lactation, respectively. Milk production showed direct relationship with buffaloes affected with post parturient hemoglobinuria.
From the present study, it is concluded thathemoglobinuria was observed in buffaloes of tehsil Bhalwal may be due to variation of soil composition particularly the deficiency of Phosphorus which may lead to the lysis of erythrocytes and hemoglobinuria through various pathways. However, efficient replenishment of minerals content in fodder producing soil is necessary to overcome the disease in buffaloes affecting from parturient hemoglobinuria in the aforementioned area.
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Pathobiological Investigations Of Peste Des Petits Ruminants (Ppr) Virus With Reference To Antiviral Activity Of Nigella Sativa (Black Seed)
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Publisher: 2017 Dissertation note: Peste des Petits Ruminants (PPR) is a highly contagious, infectious, acute or sub-acute transboundary viral disease of domestic and wild small ruminants. It is an economically important viral disease of sheep and goats causing varying degree of morbidity and mortality in susceptible animals which may be as high as 100 and 90 per cent, respectively. PPR is responsible for serious socioeconomic problems. There is no data available regarding pathogenesis and field virus characterization to compare it with vaccinal strain for any difference. Nigella sativa(Black Seed) has antiviral activity against many viruses. Therefore present studywas undertaken to investigate the antiviral effect of Black Seed in vivo and in vitro against PPR virus. Further more time course detection of virus is still needed to be studied.
Nigella sativa (Black seed) has antiviral activity against PPR virus.
Pathogenesis can better be studied through histopathology, necropcy findings and morphometric changes.
A total of 250 clinically positive samples suspected for PPR virus were included in the study. Samples were consisted of nasal, ocular and anal swabs; whole blood in EDTA were collected from suspected animals. In case of mortality morbid material included lungs, liver, spleen and mysenteric lymph nodes were included in the study. Samples were subjected to immune capture Elisa for detection of viral antigen in suspected samples. Samples which found positive foe IC – Elisa were then subjected to RT-PCR for confirmation of virus. After confirmation of virus through IC – Elisa and RT-PCR the positive samples were subjected to virus isolation on vero cell. After isolation of virus, the TCID 50 of the virus was calculated for preparation of inoculum for further use. In this experiment mesenteric lymph nodes and spleen found to be major organ for isolation of PPRV.RT-PCR found to be most reliable and confirmatory diagnostic test for PPRV. Field Virus adaptation on vero cells found to be difficult to optimize.
In this experiment antiviral activity of black seed was checked on vero cells infected with PPRV. Three extracts of N. Sativa were prepared to check the in vitro antiviral activity of black seed. In this study poly saccharides extracted from black seed found to be more effective against PPRV. Adaptation of field virus was done on Vero cell line. Antiviral activity of Black Seed extract was determined in vitro on Vero cell on bases of CPE (Cytopathic effect). The ethanolic and aqueous extract were found to be more toxic to consistency of monolayer of vero cells. The TCID50 of virus was calculated after treating cells with different extracts. In this study poly saccharides extract exhibit lower TCID50‘s as compared to ethanolic and aqueous extract which showed higher TCID50’s.So less cytopethic effect was observed in vero cells treated with black seed extracts. Antiviral activity was determined on base of CPE.
Pathogenesis of virus in natural host was studied through time course detection of virus in body secretions, blood, organs. Histopathological changes were studied.20 goats were procured from market divided into four groups (n=5) A,B,C and D. In animals of group A prophylactic effect of N.Sativa was studied. In group B complete pathogenesis of PPR virus was studied without any prophylactic or therapeutic measure. In group C therapeutic effect of N. Sativa was studied after onset of clinical picture of disease. At the end of this experiment, clinical picture, gross pathology, histopathology, and morphometric changes revealed that N. Sativa has noticeable prophylactic effect on PPR infected goats. It can be used as a therapeutic agent in PPR infected goats but it can’t control pathological effect of virus after onset of infection.
Data collected were statistically analyzed by using Microsoft Excel (Microsoft Excel, 2007) and SPSS (for Windows, Version 16.0). The data were put the descriptive analysis and Chi square test was employed to test the significance and test of hypotheses
It was concluded that Black Seed therapy possessed marvelous prophylective effect against PPR virus and RT-PCR was the most efficient methodology to confirm the virus.
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