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Detection Of Mycoplasma Synoviae By Pcr And Its Histopatholohical Studies In Poutry Breeder In District Abbottabad

By: Sajjad Ahmad | Dr. Muti- ur- Rehman Khan.
Contributor(s): Prof. Dr. Muhammad Younus Rana.
Material type: materialTypeLabelBookPublisher: 2011Subject(s): Department of PathologyDDC classification: 1316,T Dissertation 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.
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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.

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