1.
Detection Of Mycoplasma Synoviae By Pcr And Its Histopatholohical Studies In Poutry Breeder In District Abbottabad
by Sajjad Ahmad | Dr. Muti- ur- Rehman Khan | Prof. Dr. Muhammad Younus Rana.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2011Dissertation note: Poultry, an important sub-sector of livestock, has emerged a cheaper source of protein for human consumption. Mycoplasmas are the smallest known bacteria, 300-800 nm in diameter and are capable of replicating outside the cell. Mycoplasma synoviae is a member of the class Mollicutes, order Mycoplasmatales, family Mycoplasmataceae. Mycoplasma synoviae (MS) is considered economically to be most important pathogen. Mycoplasma synoviae infections occur in poultry worldwide, affecting poultry and causes diseases like respiratory distress, synovitis and arthritis. Mycoplasma is transmitted from infected to healthy birds both by horizontal and vertical routes. Horizontally disease is transmitted via infected and healthy carrier birds, hatchery, housing, equipments, feeding and during transportation.
To have an insight on pathogenesis and reliable diagnostic techniques, the present project was designed to know comparative sensitivity of rapid agglutination test and polymerase chain reaction for MS diagnosis and to study the gross lesion and histopathological changes in chicken joints produced by MS.
The birds showing clinical signs that included respiratory i.e. tracheal rales, conjunctivitis, coughing, sneezing, ocular and nasal discharge and infectious synovitis were selected for sample collection. Initially the collected sera samples were examined by Rapid Serum Agglutination test. RSA and PCR tests were used in order to confirm the pathogenic agent. RSA and PCR positive samples were further processed for histopathological study in order to identify the lesions in tissues produced by causative organism. In field visits it was observed that the suspected birds were with pale comb, mild to severe lameness, dull, depressed, ruffled feather, conjunctivitis, oculo-nasal discharge, tracheal rales and greenish or sulfur faeces. Birds hock joints, toe joints and paws pad were swelled. The infected birds were occasionally found with generalized infection. The infected birds complicated with other diseases of poultry such as Newcastle and infectious bronchitis causes infection airsacculitis.
Rapid serum agglutination test was conducted at 14 broiler breeder farms. The birds at a farm were showing respiratory and infectious synovitis signs and symptoms, suspected to Mycoplasma synoviae. The tests were performed at the spot. A total of 239 sera samples were examined out of which 63 (26.35%) sera samples were positive for MS. The clinical samples were identified and confirmed as Mycoplasma synoviae infection by PCR. The amplified PCR product was given about 211 bp size while PCR buffer was used as negative control. A total of 213 samples were subjected to PCR and 65 (30.52%) revealed PCR positive results for tracheal swabs, 28.16% (20 samples out 71) showed positive results. For tracheal and lung 33.38 % (24 out of 71) and 29.57% (21 out of 71 samples) were positive, respectively. The PCR test successfully amplified the DNA of MS clinical positive samples. Sixty five out of 213 Mycoplasma synoviae isolates were positive in MS specific PCR while the other 148 samples were negative. The sensitivity and specificity of molecular method Polymerase chain reaction was 100 percent.
For histopathological studies the samples of different organs including trachea, lungs, liver, hock joints (articular cartilage, piece of synovial membrane) and foot pad were further processed. The trachea was examined. There was epithelial degeneration, desquamation. congestion, haemorrhages and inflammatory cell infiltration. The lungs were examined and it was revealed that there was marked congestion, haemorrhages, necrosis and mononuclear cells infiltration. Liver showed infiltration of lymphocytes, plasma cells and macrophages. Articular cartilage showing chondrocytes degenration. Synovial membrane was thickened due to infiltration of lymphocytes and plasma cell. Foot pad showed hyperkaratosis and thickning of epidermis, acanthosis, degeneration of cartilage, infiltration of both mononuclear and plasma cell.
It is concluded from findings of present study that PCR is more appropriate technique than RSA for timely diagnosis of Mycoplasma synoviae. However combination of findings of both techniques may be utilized for accurate detection of Mycoplasma synoviae from broiler breeder in district Abbottabad.
Availability: Items available for loan: UVAS Library [Call number: 1316,T] (1).
2.
Histopathological Investigation Of Pleuropneumonia In Buffaloes Caused By Mycoplasma Bovis
by Ayesha Rabail | Dr. Muti-Ur-Rehman Khan | Dr. Kamran | Prof. Dr. M Younus Rana.
Material type: ; Format:
print
Publisher: 2009-2011Dissertation note: This study was conducted by keeping in view the worldwide importance of Mycoplasma bovis to cause pneumonia and many other diseases, as it causes great economic losses to bovine industry. In the current project the incidence of Mycoplasma bovis to cause pleuropneumonia was studied, and its respective histopathological changes in lungs of the pneumonic adult buffaloes and buffalo calves were examined. 100 lung samples for this purpose (50 lung samples from adult buffaloes and 50 lung samples from buffalo calves) were collected from the Lahore Bakar Mandi Abbatoir. Samples were collected on the basis of following criteria: Red hepatization, grey hepatization, multifocal abscess, necrotic lung tissue. These samples were then divided into two portions, one half placed in 10% buffered formalin in the bottles and other half kept in sterile polythene bag. The portion of lungs for bacteriological study was kept in ice box. Histopathological procedure was performed in the pathology department of University Of Veterinary And Animal Sciences Lahore. The samples were subjected to histopathological procedures and then slides were observed microscopically for the changes. Microscopically pulmonary odema, consolidation, caseous necrosis, abscess infiltration of mononuclear cells, plasma cells, macrophages, neutrophils infiltration were observed. For culturing of Mycoplasma bovis PPLO broth was prepared and samples were inoculated in the broth medium. At 7th day of inoculation the yellow color of the broth medium appeared which was indicative of positive samples. 30% positive samples in adult buffaloes and 36% in buffalo calves were obtained. These samples were then inoculated on the PPLO agar plates for further precision of results. On agar plates typical colonies of the Mycoplasma were observed under bright field compound microscope and 60% positive samples in adult buffaloes and 66% in buffalo calves were obtained. Next step towards the confirmation of Mycoplasma bovis was specific acridine staining, in which positive of Mycoplasma bovis samples gave dull yellow to colorless appearance of yellow broth medium and gave egg fried colony on agar. 78% adult buffalo and 67% buffalo calves showed positive results. These samples were then subjected to final confirmatory test which was growth inhibition disk test, in which hyper immune sera was raised in rabbits and filter paper disks soaked in this sera were used to check the zone of inhibition on cultured agar plates. 70% positive samples in adult buffaloes and 75% in buffalo calves were obtained which confirmed the presence of Mycoplasma bovis. CFU/ml of the positive samples calculated between 105-108. So the incidence of Mycoplasma bovis to cause pneumonia in adult buffaloes and buffalo calves calculated was (10% and 12%) respectively.
Availability: Items available for loan: UVAS Library [Call number: 1335,T] (1).
3.
Effect Of Zinc Supplementation On Cadmium (Cd) Toxicity In Japanese Quail (Coturnix Coturnix Japonica)
by Umair Ishtiaq | Prof. Dr. Muhannad Younus Rana | Dr. Muti - Ur- Rehman Khan | Prof. Dr.
Material type: ; Format:
print
Publisher: 2010Dissertation note: Cadmium (Cd) has been recognized as an environmental pollutant especially due to its anthropogenic activity. Exposure to Cd is known to cause harmful effects of different levels of the trophic chain, because of bioaccumulation. Toxic effects of Cd are observed in the kidneys, lungs, testes, prostate, and the erythropoietic system in chicken and rat. These effects are associated with teratogenesis and carcinogenesis. Zinc posseses protective and antagonistic action in the uptake and toxic effects of cadmium, probably because of Zn-induced synthesis of metallothionein that detoxifies Cd by firmly binding this metal. The objectives of present study are:
1) To understand the effect of zinc on cadmium toxicity in Japanese quails
2) To study the effect of zinc and cadmium on liver and kidney function along with histopathological changes.
This experimental trial was carried out at Avian Research and Training Centre (ARTC), UVAS Lahore and tests were performed at Pathology Deptt UVAS Lahore. Day old Japanese quails (C. coturnix japonica) (n = 560) were used in this experiment. For this purpose, a total number of 560, day old chicks of Japanese quail were procured from the hatchery of ART Centre. They were assigned to seven dietary treatments. There were four replicates in each treatment group and each replicate was of twenty chicks.
A =Control diet group will receive only basal diet without any supplementation. B= Basal diet + 50 mg/kg Cd, C = Basal diet + 75 mg/kg Cd, D =Basal diet +100 mg/kg Cd E=Basal diet+50 mg/kg Cd+40 mg/kg Zn, F=Basal diet+75 mg/kg Cd + 40mg/kg Zn and G =. Basal diet+100 mg/kg Cd + 40mg/kg Zn. Each group consisted of 80 birds. The body weight of each the birds was carried out weekly. Blood samples were collected on the 21st (3rd week), 28th (4th week), 35th (5th week) and 42nd (6th week) days of treatment from each group to evaluate the liver and kidney functions and vital organs were collected after slaughtering from each group and then histo-pathological analysis was done. The data was analyzed by two way ANOVA.
Availability: Items available for loan: UVAS Library [Call number: 1339,T] (1).
4.
Study On The Pathogenesis Of Clostridium Perferingens (Necrotic Enteritis) In Experimentally Infected
by Arif Mehmood | Dr. Muti-ur-Rehman Khan | Prof. Dr | Prof. Dr. M. Younus Rana.
Material type: ; Format:
print
Publisher: 2011Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1546,T] (1).
5.
Pathology Of Experimental Enterotoxemia In Sheep And Goats
by Azam Ali Nasir | Prof. Dr. M. Yonus Rana | Dr. Muhammad | Dr. Muti-ur Rehman Khan.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation note: During the present study intestinal scrapings were collected from sheep suspected for enterotoxaemia. Samples were subjected for the isolation by repeated culturing in Reinforced Clostridium medium. Growth on blood agar revealed characteristic colonies of Clostridium perfringens after 18-24 hours. Biochemical and mice inoculation tests were performed. Isolated organism was identified by indirect ELISA. The pure growth was subcultured in RCM in bulk quantity and calculated the dose for experimental infection.
In the second part of the study, sheep and goats were procured, dewormed and kept in the experimental house of UVAS, Lahore. The experimental infection comprised of the whole culture of C. perfringens type D was inoculated intraduodenally via para-mid line between animals of group A and B while in animals of group C and D inoculated only starch solution to achieve the objectives. Accumulative clinical score in sheep was found to be 9 to 16, 13 to 22 and 15 to 23 at 10, 20 and 30 hours PI respectively while in goats the accumulative scores varied from 5 to 15, 9 to 16 and 14 to 21 at above mentioned time intervals. The highest mean score for clinical findings in sheep was anorexia, frothing followed by dehydration while in goats, the highest mean score was recorded for diarrhea dehydration, and anorexia. No significant clinical findings were noted in control groups.
There was a significant increase in blood glucose, urea and serum creatinine in infected group of sheep and mean values reached up to 141 mg/dl, 92 mg/dl and 7.5 mg/dl respectively at 30 hours PI while in goats a similar pattern was observed with the mean values raised to 142 mg/dl, 111 mg/dl and 10.2 mg/dl for blood glucose, urea and serum creatinine respectively. There was no significant change found in RBC and platelet count of both species but there was an early increase in the mean WBC count of sheep 19.7x103/µl at 10 hours but then decreased to 14.7x103/µl at 30 hours PI while in goats it was 23.6x103/µl and then decreased to 15.3x103/µl. The mean PCV % age increased in animals of both infected groups but more in goats and reached to 52% in 30 hours.
During the third part of the present study, the animals were slaughtered and PME performed. The accumulative score for gross lesions were recorded and it was found between 14 to 24 in animals of group A with the highest score for congestion and edema of different organs whereas in goats it was between 12 to 22 with congestion and hemorrhages of intestine having highest scores. The samples were kept in formalin for histopathological examination and accumulative lesions score was noted in different organs. The highest mean score in sheep was recorded in kidneys and lungs and in goat intestine, lung and kidneys were the major organs affected.
A polymerase chain reaction was optimized under our own laboratory condition for the detection of alpha and epsilon toxins of Clostridium perfringens type D from different tissues. Alpha gene was amplified at annealing temperature 52.2oC with amplicon size 247bp and ETX gene at the annealing temperature.
50.2oC with amplicon size 665bp.
The erythrocytes of different species were used to know the sensitivity against culture supernatants of C. perfringens type D. It was observed maximum hemolysis occurred in human erythrocytes (68%) followed by mice (57%) at 37oC. It was also recorded that a significant increase was found at 37oC as compared to25oC except for dog and rabbits where no significant difference was observed.
Availability: Items available for loan: UVAS Library [Call number: 1596,T] (1).
6.
Clinicopathological Study Of Theileriosis In Naturally Infected Sheep
by Muhammad Waseem Akhter | Dr. Asim Aslam | Dr. Muti ur Rehman Khan | Prof. Dr. Habib.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2013Dissertation 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.
Availability: Items available for loan: UVAS Library [Call number: 1642,T] (1).
7.
A Study On Immunomodulatory Effect Of Nigella Sativa And Allium Sativum Against Escherichia Coli Infection In Broiler Birds
by Fiza Ahmad | Dr Muhammad yasin tipu | Dr. Muti-ur- rehman khan | Prof. Dr.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2022,T] (1).
8.
Esponse Of Lymphatic Tissues To Dietary Supplementation Of Nigella Sativa And Curcuma Longa In Broilers Challenged With Pasteurella Multocida (strain A)
by Muhammad Akmal raheem | DR. Muhammad Yasin tipu | Dr. Muti ur rehman khan | Prof. Dr.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2050,T] (1).
9.
Pathological Investigations Of Theileriosis (T.Annulata) In Cattle In Disteict Lahore Punjab
by Syed Sadeed ud din Shah | Dr. Muti-ur- Rehman Khan | Dr. Raheela Akhtar | Prof. Dr. Aneela Zameer Durrani.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Theileriosis is very important protozoal disease in crossbred cattle. According to an
assessment, about 250 million cattle are endangered by this disease and millions of high milk
yielding cattle are at risk of exposure to disease. It acts as a bigger restraint on livestock
improvement and production in many developing countries (Nagore et al. 2004). Theileria
annulata is the main specie that causes high morbidity and mortality. It causes heavy economic
and production losses in cattle in tropical and sub-tropical regions. The recorded mortality rates
in cattle reached to 70% (Moor house et al. 2001).
Theileria species are intracellular obligate hemoprotozoan parasites. All Theileria species
are dangerous and cause disease but two of them are important for livestock. Theileria parva and
T. annulata produces diseases named as East Coast fever and tropical theileriosis in cattle
respectively. Genus Theileria has many other species like T. buffeli, T. taurotragi, T. velifera, T.
sergenti and T. mutans. These species cause infections in wild and domesticated ruminants.
Theileria species present in large and small animals show signs like fever, anorexia,
swelling of the superficial lymph nodes, dyspnoea, lethargy, progressive anemia, constipation,
diarrhea, lacrimation and nervous symptoms (Saeed et al. 2010; Irvin and Mawmachi 1983). A
pronounced rise in body temperature, reaching 40-41.5 °C is pursued by lacrimation, depression,
swelling of the superficial lymph node and nasal discharge. The characteristic sign of tropical
theileriosis is anemia and finally haemoglobinuria occur with heavy weight losses. The clinical
course of the disease alter from per acute to acute or sub-acute to chronic (Oliveira- Sequeira et
al. 2005). The disease is lymphoproliferative in its early phases resulting enlargement of lymph
nodes, later on enters lymph destructive phase which is associated with a pronounced
Introduction
2
leukopenia. In the piroplasms phase in erythrocytes, the parasite becomes infective for the tick
(El-Deeb and Younis 2009).
Trans placental Bovine Tropical Theileriosis causing a deadly disease in a 3 day old
neonate cross bred calf and cerebral form of the disease (turning sickness) in a cow were
incriminated to T.annulata infection. It mainly depends upon the harmful effects of the T.
annulata on lymphoid tissues and susceptibility of the host (Sudan et al. 2012).
Theileriosis is prevalent in various regions of the world including Pakistan. It is transmitted by
Hyalomma species ticks. These ticks spread T. annulata which causes tropical theileriosis
(Durrani et al. 2009). The developmental stages of Theileria inside the Hyalomma ticks varies in
different shapes and forms (Hamed et al. 2011). Therefore to increase the milk and meat
production of cattle we can prevent the spread of the disease by controlling ticks
(Hekmatimoghaddam et al. 2012). The sufficient amount of Hyalomma ticks are found in warm,
commonly hard marshland and in central and Southern Europe, south west Asia and Southern
Africa having very long dry season. A toxin is produced in the adult ticks. This toxin produce
clinical signs of mucus membrane hyperemic and moist profuse eczema (Adam et al. 2000).
The sporozoites of Theileria enter into cattle host during tick feeding and they
immediately infect mononuclear leukocytes, these sporozoites develop into macroschizonts and
induce proliferation of the host cells. Macroschizonts constantly mature into microschizonts and
finally into merozoites, which are discharged from leukocytes. These merozoites attack
erythrocytes and mature into piroplasms, become available to ticks. Infective sporozoites,
injected during tick feeding, rapidly enter target cells, escape from the surrounding host-cell
membrane and differentiate to schizonts that interact with different host-cell components
(Dobbelaereand Rottenberg 2003). This interaction includes host cell signaling pathways that
Introduction
3
regulate proliferation and cell survival (Chaussepied and Langsley 2011) and thus cause
blastogenesis and clonal expansion of predominantly T and B cells (Fawcett et al. 1982; Baldwin
et al. 1988; Spooner et al. 1989). Merozoites released from these schizonts subsequently infect
red blood cells and become trophozoites. Lymphocytic stage of Theileria (schizonts) is the cause
of many of the severe disease manifestations like lymphadenopathy, pyrexia, thrombocytopenia,
and panleukopenia (Homer et al. 2000). Marked anemia, anisocytosis, pikilocytosis, and
leucopenia were commonly observed in bovine theileriosis (Ceci et al. 1997). Cattle may survive
the disease, but recovery and convalescence may be protracted and incomplete, this leads to
permanent debilitation, loss of productivity and prolonged carrier state. (Shahnawaz et al. 2011).
Cattle with subclinical infection in endemic regions become carrier of piroplasms and act as a
source of infection for the vectors (Brown 1997; Brown 1990; Uilenberg 1995).
The diagnosis of theileriosis in acute cases is majorly done on clinical signs and Giemsa
stained blood smears of cattle but the detection of agent is not reliable and is almost impossible
in carrier stage. Advances in molecular biological techniques have resulted in the improved
detection, identification, and genetic characterization of many hemoparasites. Species specific
polymerase chain reaction (PCR) has been developed for the detection and identification of
various Theileria species and has been shown to have higher sensitivity and specificity compared
with serological assays and examination of Giemsa-stained blood smears (Bhoora et al. 2009).
Primers were derived from the gene encoding the 30-kDa major merozoite surface antigen for T.
annulata (Aktas et al. 2006).
Most of the previous studies on haematological parameters in T. annulata infection were
carried out on experimentally infected cattle (Sandhu et al. 1998; Singh et al. 2001). The present
investigation was conducted to study haematological parameters in cattle naturally infected with
Introduction
4
T. annulata. Hematology has been broadly used in attempts to give information about disease
condition, performance problems and health in cattle (Rezaei and Naghadeh. 2006).
Hematological and sero-biochemical alterations are the indicators of severity of disease and are
considered to be good tools for the diagnosis, prognosis for effective therapy (Col and Uslu
2007; Nazifi et al. 2010b).
Lahore is one of the larger district in the Punjab province of Pakistan. Different cattle
breeds are reared by the people of the area for meat and milk production. The exact current
situation about the prevalence and pathogenesis of Theileriosis in the selected area is unknown.
The present study was conducted to screen cattle by finding schizonts or piroplasms in Giemsa
stained thin blood smears at slaughter house of district Lahore (Aktas et al. 2006) and later to
confirm through Polymerase Chain Reaction (PCR) (Chaisi et al. 2013) in order to implement
efforts and regulation to eradicate the spread of disease in the selected area. Data generated from
this study provided the latest status of Theileriosis, sex wise prevalence and its pathogenesis in
cattle population of Lahore. The study has also provided the necessary information to formulate
strategies for control of disease in the area. An investigation was also undertaken to ascertain the
changes in haematology as a result of Theileria annulata infection. These studies will help better
understanding of the pathogenesis and supportive therapy of this disease. Availability: Items available for loan: UVAS Library [Call number: 2186,T] (1).
10.
Pathobiological Studies On Bovine Ephemeral Fever Infected Cattle In District Swabi
by Sahibzada Waheed Abdullah (2013-VA-560) | Dr. Muti Ur Rehman Khan | Prof. Dr. Asim Aslam | Dr. Ali Ahmad Shiekh.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation 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.
Summary
31
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. Availability: Items available for loan: UVAS Library [Call number: 2284-T] (1).
11.
Study On The Pathogenesis Of Co-Infection Of Infectious Bronchitis (Ibv) And Escherichia Coli (E. Coli) In Experimental Chickens
by Sohail Khan (2013-VA-606) | Prof. Dr. Asim Aslam | Dr. Muti Ur Rehman Khan | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation 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,
Summary
56
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. Availability: Items available for loan: UVAS Library [Call number: 2306-T] (1).
12.
Pathological Investigation And Molecular Detection Of Avian Pathogenic E.Coli Serogroups In Broiler Birds
by Muhammad Azeem Riaz (2008-VA-132) | Prof. Dr. Asim Aslam | Dr. Muti Ur Rehman Khan | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation 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.
Summary
46
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. Availability: Items available for loan: UVAS Library [Call number: 2591-T] (1).
13.
Study On The Pathogenesis Of Field Isolate Of Salmonella Pullorum In Experimentally Infected Broiler Chicken
by Muhammad Zeeshan (2014-VA-535) | Dr. Muti-ur-Rehman Khan | Dr. Gulbeena Saleem | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Poultry is one of the well organized sectors in Pakistan and generates employment for up
to 1.5 million people. As use of modern techniques and medicine has reduced a risk lot but it is
still facing threat from many diseases. One of these is Salmonellosis which is and hatchery borne
infection and it also has a zoonotic importance. This organism is gram negative bacterium, non
acid fast, uniform staining and non spore forming bacillus.
Pullorum disease produced by it is also known as bacillary white diarrhea due to whitish
droppings produced by birds. Day old chicks are affected because it can be transmitted vertically.
Infected fomites and utensils also play a role in the transmission of infection. Growth of S.
pullorum rapidly occurs upon beef agar on nutrient media and growth is best at 37 °C. Selective
media such as selenite, tetrathionate broths and differential media such as MacConkey, bismuth
sulfide and brilliant green agar are best for the proper growth of organism.
The organism was isolated from the 30 different infected samples of broiler birds taken
from different farms and was grown upon the selective media. The colonies were picked from
the cultures and were subjected to PCR for the further process. After confirmation with PCR,
inoculums was prepared with a bacterial load of 2 x 107 (CFU) in 0.5 ml of normal saline at the
ph of 7.2.
Chicks were randomly divided in to three groups as group 1, 2 and 3, each group
containing 30 chicks. Group 1 was infected with field strain of S.pullorum, Group 2 with
vaccinal strain and Group 3 was set as control group and no infection was given to it. Samples
were collected from birds at 10th, 15th, 20th and 25th days of age. Observations were made upon,
clinical signs, histopathological lesions, mortality and morbidity rate, FCR and gross
pathological lesions.
Clinical signs included were anorexia, depression, huddling, labored breathing, loss of
feed and water intake, ruffled feathers, reduced mean body weights, low FCR and pasty vent.
Gross and histopathological lesions were much observed and were prominent in the birds
affected with field strain of S. pullorum. One Way-ANOVA was used for statistical analysis in
between the groups. Gross pathological lesions included were liver congestion 55%, cheesy
material in caeca 70% in the birds which were affected with field strain of S. pullorum.
Histopathological findings in the birds affected with field strain were monocytes in spleen,
leukocytes in intestine and heterophils in liver. Low FCR was recorded in infected birds of
Group 1 and was 1.63 at 25th day. Mortality rate in the birds affected with field strain was
maximum 43.33% and morbidity rate was 73.33% at the 10th day of age. Birds of group 2 were
less affected and only showed medium level of lesions. No specific lesions were seen in group 3.
The total prevalence of of S.pullorum after conformation with PCR was 20% out of total 30
infected samples. The organs for histopathology taken were liver, spleen, caeca and intestine and
were preserved in 10% formalin prior to the infection.
The overall pathogenesis of experimental bacteria after given through oral route was that
it first localizes in digestive tract from where it enters into the blood stream and invades in the
different organs and tissues at different time intervals producing lesions and immunological
response. Availability: Items available for loan: UVAS Library [Call number: 2693-T] (1).
14.
A Study On Point Prevalence, Etiological And Biochemical Investigations Of Post Parturient Haemoglobinuria In Buffaloes In Tehsil Bhalwal
by Muhammad Azeem (2015-VA-430) | Dr. Muti-ur-Rehman Khan | Prof. Dr. Asim Aslam | Prof. Dr. Shafqat Fatima Rehmani.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation 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.
Availability: Items available for loan: UVAS Library [Call number: 2847-T] (1).
15.
Pathological Effects Of Natural And Experimental Lead (Pb) Toxicity In Lohi Sheep At Jhang, Pakistan
by Muhammad Sajid (2010VA-61) | Prof. Dr. Muhammad Younus | Dr. Muti ur Rehman Khan | Prof. Dr. Aftab Ahmed Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Heavy metal toxicity is increasing due to increasing trends of urbanization and
industrialization. Lead poisoning has been recognized as a major public health risk,
particularly in developing countries. It is nephrotoxic, hepatotoxic, neurotoxic, carcinogenic
and mutagenic for animals and human. Sewerage water, fertilizers, leaded-gasoline and lead
based batteries are the sources of lead contamination in soil and forage. The lead particles are
taken up by animals from contaminated forages and excreted in animal products like milk and
meat.
The presence of Pb in drinking water, waste water, plant products and animal
products has been studied which is a serious risk for animal and public health. The
legislations for the disposal of household wastes and industrial effluents are very poor in
Pakistan. The calculation of safe Pb levels in different products is still to be needed.
Pathological effects of higher Pb levels have not been studied in Pakistan. The present study
was aimed to unveil the toxic effects at constant dose of Pb over a period of three months in a
local sheep breed of Pakistan. The status of Pb toxicity was also investigated in a polluted
area around sewage drain and mutton slaughter house at District Jhang, Pakistan.
The Pb concentration in soil, forage and irrigating water was found to be below the
permissible limits and was safe for agriculture but long-term ingestion of low Pb
concentration may have cumulative effect. The serum Pb concentration was found to be
above the recommended safe limits for producing Pb toxicity in animals. The different tissues
like kidney, liver and skeletal muscles also contained higher Pb level from the permissible
limits and found to be unsafe for public use. Kidney showed the highest Pb concentration and
the muscle contained the least Pb level in the present study.
Summary
142
The erythrocyte count, hemoglobin concentration and packed cell volume showed
inverse correlation with Pb concentration and mean values were below the normal range in
Pb treated sheep but anemia was not developed. The erythrocyte sedimentation rate was also
influenced by given dose of lead acetate during third month of treatment. The white blood
cells also revealed no effect on given dose of lead acetate in Lohi sheep in this study.
The biochemical parameters of field and treatment group showed higher concentration
as compared to control group of Lohi sheep but their means were falling within the normal
range of reference values. The disturbed biochemical parameters in apparently healthy sheep
with higher serum Pb concentration were indicative for liver and kidney damage.
Lohi sheep exhibited less effect on given dose of lead acetate during first two months
but more pronounced changes of chronic Pb toxicity were observed during last month of trial.
The histological changes were not observed on early period in lead acetated treated sheep.
The characteristic histological changes were observed on last slaughtering at day 90 in kidney
and liver including degeneration and focal areas of necrosis, dilatation of blood vessels with
accumulation of red blood cells and fibrosis in some areas. The nuclear changes were more
typical with intranuclear inclusion bodies in renal tubular epithelial cells but less
distinguishable in hepatocytes.
It was concluded that soil, forage and water contained low Pb levels in the study area.
The ingestion of low Pb level for longer period had cumulative effect in animals. The animals
might be resistant to low Pb level but their products are a severe risk for public health. So the
necessary measures should be adopted to minimize the heavy metal contamination in animal
products. Availability: Items available for loan: UVAS Library [Call number: 2820-T] (1).
16.
Pathobiological Investigations Of Peste Des Petits Ruminants (Ppr) Virus With Reference To Antiviral Activity Of Nigella Sativa (Black Seed)
by Kiran Aqil (2008-VA-456) | Dr. Muti Ur Rehman Khan | Prof. Dr. Asim Aslam | Dr. Aqeel Javeed.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation 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.
SUMMARY
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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. Availability: Items available for loan: UVAS Library [Call number: 2890-T] (1).
