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Isolation and Molecular Characterization of Mycoplasma gallisepticum and Mycoplasma synoviae in Chickens in Sudan  [PDF]
Khalda A. Khalifa,Egbal Sidahmed Abdelrahim,Magdi Badwi,Amal M. Mohamed
Journal of Veterinary Medicine , 2013, DOI: 10.1155/2013/208026
Abstract: The current study described the isolation and molecular detection of Mycoplasma gallisepticum (Mg) and Mycoplasma synoviae from tracheal swabs of diseased birds showing signs of respiratory distress in selected commercial (layer and broiler) farms and from yolk and an open air of pens of vaccinated breeder flocks in Sudan. A number of 45?Mycoplasma isolates were recovered from chickens in Khartoum, Gezira, and Equatoria states in Sudan. Of these, eight Mg and three Ms isolates were identified using growth inhibition and rapid serum agglutination (RSA) tests. The conventional PCR technique was applied to amplify 140?bp and 720?bp DNA fragments for the Mg and Ms, respectively. This research confirmed vertical and horizontal transmission of Mg from breeder farms through detection of Mg in yolk of fertile eggs and an air of pens despite previous vaccination. PCR is considered a rapid, sensitive, and cheap method and it will improve the diagnosis of Mycoplasma in chickens. 1. Introduction Avian mycoplasmosis was primarily described in turkeys in 1926 and in chickens in 1936 [1]. Mycoplasma gallisepticum (Mg) infection is usually designated as chronic respiratory disease of chickens and infectious sinusitis in turkeys. It is characterized by respiratory rales, coughing, nasal discharges, and frequently by sinusitis in turkeys by synovitis. Mycoplasma synoviae (Ms) infection is usually known as infectious synovitis, an acute-to-chronic infectious disease for chickens and turkeys involving primarily the synovial membranes of joints and tendons sheaths. However, during recent years, Ms has less frequently been associated with synovitis but more frequently associated with airsacculitis in chickens and sometimes in turkeys [2]. Both diseases are economically important, egg transmitted and hatchery disseminated diseases. They lead to tremendous economic losses in poultry production as a result of decreased hatchability and egg production, reduced quality of day-old chicks, reduced growth rate, increased costs of eradication procedures which involve site cleaning and depopulation, and increased costs of medication and vaccination [3]. The first isolation of both mycoplasmas in Sudan was reported by Khalda [4]. A recent study indicated that these organisms were prevalent, as 50.8% Mg and 57.6% Ms antibodies were recorded in chickens in the country [5]. For many years, diagnosis of avian mycoplasmosis was based on serological assays to detect antibody production and/or on isolation and identification of the organism. Serological tests include the rapid slide
Serological investigation of five diseases; Influenza, Newcastle disease, Salmonella, Mycoplasma gallisepticum and Mycoplasma synoviae in native hens of Eghlid, Iran  [cached]
A. Shadmanesh,M. M. Mokhtari
Veterinary World , 2013, DOI: 10.5455/vetworld.2013.126-130
Abstract: Aim: The study was conducted to determine seroprevalence of the five diseases influenza, Newcastle, Mycoplasma gallisepticum, Mycoplasma synoviae and salmonella, among around native hens of Eghlid in Iran, on spring 2011. Materials and Methods: On the basis of native Hens distribution, this region divided into four parts of Eghlid, Doskord, Sedeh and Hasan-abad. Fifty unvaccinated native Hens randomly selected from each part. Blood samples were aseptically collected from the wing veins using 5-ml sterile syringe. Serum from hens was tested for detection and titration for Mycoplasma and Salmonella by the rapid slide agglutination method, and was tested for influenza and Newcastle by the Hemagglutination Inhibition Assay. The data was analyzed completely in randomized design with four treatments, 50 repetitions for each disease. Results: 34 out of 200 samples (17%) were positive for influenza. There were significant differences between regions (p<0.01). 38 out of 200 samples (19%) were positive for Newcastle. The maximum infectious rate obtained from Eghlid. There were significant differences between regions (p <0.05). 170 out of 200 samples (85%) were positive for Mycoplasma gallisepticum. 4 from 200 samples (2%) were positive for Mycoplasma synoviae. The results do not show a significant difference for salmonella (p <0.05). Conclusion: Contamination of Influenza, Newcastle and Mycoplasma gallisepticum was high, and the highest contamination rate was related to Mycoplasma gallisepticum. It is usually recommended that preventive strategies, such as appropriate husbandry and hygiene, sanitary handling of chicks and eggs, routine health monitoring and vaccination of Native hens should be emphasized. [Vet World 2013; 6(3.000): 126-130]
Relative Risk Estimation for Mycoplasma synoviae in Backyard Chickens in Paraguay  [PDF]
K. Suzuki,J. Origlia,F. alvarez,M. Faccioli
International Journal of Poultry Science , 2009,
Abstract: Poultry production is a growing industry in Paraguay, in southern South America. The insufficient farm management methods frequently applied in backyard chickens make them a possible reservoir for economically important diseases such as Mycoplasma synoviae that can influence commercial poultry operations. There are no former studies on a survey of Mycoplasma synoviae among backyard chickens in Paraguay. The objectives of this study were: (1) to observe the seroprevalence of MS in backyard chickens in Paraguay and (2) to generate maps for the estimated Relative Risk (RR) for MS in the study chickens, in place of using the observed seroprevalence. Paraguay is divided into 17 departments. A department-stratified random sampling was arranged and conducted. The required total sample size of 1537 from a chicken population of 17 million was sufficient to produce a 95% confidence interval with a desired precision of ±2.5% when the estimated seroprevalence was 50%. Sera were examined using a commercial indirect ELISA. The overall observed seroprevalence was 53%. The resulting maps for the estimated RR for Mycoplasma synoviae in the study chickens at department level were drawn. Departments with notably high or low disease risks were confirmed. Different types of epidemiological parameters can be calculated to take account of probable risk factors. Therefore, additional detailed investigations into those risk factors relating to Mycoplasma synoviae occurrence with respect to spatially epidemiological dissimilarities would be of interest.
Extracellular Proteins of Mycoplasma synoviae  [PDF]
Manuel Sebastián Rebollo Couto,Catia Silene Klein,Daiane Voss-Rech,Hernán Terenzi
ISRN Veterinary Science , 2012, DOI: 10.5402/2012/802308
Abstract: Mycoplasma synoviae is a Gram positive bacteria lacking of cell wall that affects chickens and turkeys causing infection in the upper respiratory tract and in some cases arthritis, with economical impact to broiler breeders. Treatment and prevention of avian synovitis depend on knowledge of the infectious process. Secreted or surface-exposed proteins play a critical role in disease because they often mediate interactions between host and pathogen. In the present work, we sought to identify possible M. synoviae secreted proteins by cultivating the bacteria in a modified protein-free Frey medium. Using this approach, we were able to detect in the cell-free fraction a number of proteins that have been shown in other organisms to be secreted, suggesting that they may also be secreted by M. synoviae. 1. Introduction The growth of poultry industry is often limited by infectious diseases that affect birds. Mycoplasma synoviae is a major avian extracellular pathogen associated with synovitis in chickens and turkeys [1, 2]. Disease can occur as chronical subclinical to severe upper respiratory infection and, under unknown conditions, become systemic and cause arthritis [3]. The disease causes economic losses by retarding growth and downgrading at slaughter [3]. Strategies to control this pathogen rely mainly in better management practices, improvement in housing conditions and antibiotic usage, whereas an effective vaccine is still not available [4]. Secreted proteins of pathogenic bacteria are key factors in host colonization. The analysis of these proteins, called secretome, can therefore permit the identification of new putative virulence factors that are fundamental for host invasion and survival in the environment within the host [5]. In this context, two-dimensional electrophoresis (2DE) along with peptide fingerprinting by mass spectrometry (MS) and subsequent protein identification have become a powerful method to unravel pathogenicity factors in microorganisms [6, 7]. We have recently reported a proteomic analysis of M. synoviae cell extracts in conventional Frey medium [8]. In the present work, we have grown M. synoviae in the same typical culture medium and then incubated the cells in a protein-free modified Frey medium as a strategy to indicate proteins that can be secreted to the medium by the bacteria. 2. Methods 2.1. Mycoplasma synoviae Cultures M. synoviae strain 53 isolated from a broiler breeder was grown in the Laboratory of Genetics and Animal Health from EMBRAPA Swine and Poultry (Concórdia, C, Brazil) as described by Frey and coworkers
Identification of the GTPase superfamily in Mycoplasma synoviae and Mycoplasma hyopneumoniae
Borges, Clayton Luiz;Parente, Juliana Alves;Pereira, Maristela;Soares, Célia Maria de Almeida;
Genetics and Molecular Biology , 2007, DOI: 10.1590/S1415-47572007000200007
Abstract: mycoplasmas are the smallest known prokaryotes with self-replication ability. they are obligate parasites, taking up many molecules of their hosts and acting as pathogens in men, animals, birds and plants. mycoplasma hyopneumoniae is the infective agent of swine mycoplasmosis and mycoplasma synoviae is responsible for subclinical upper respiratory infections that may result in airsacculitis and synovitis in chickens and turkeys. these highly infectious organisms present a worldwide distribution and are responsible for major economic problems. proteins of the gtpase superfamily occur in all domains of life, regulating functions such as protein synthesis, cell cycle and differentiation. despite their functional diversity, all gtpases are believed to have evolved from a single common ancestor. in this work we have identified mycoplasma gtpases by searching the complete genome databases of mycoplasma synoviae and mycoplasma hyopneumoniae, j (non-pathogenic) and 7448 (pathogenic) strains. fifteen orfs encoding predicted gtpases were found in m. synoviae and in the two strains of m. hyopneumoniae. searches for conserved g domains in gtpases were performed and the sequences were classified into families. the gtpase phylogenetic analysis showed that the subfamilies were well resolved into clades. the presence of gtpases in the three strains suggests the importance of gtpases in 'minimalist' genomes.
Vaccination with newcastle disease vaccines strain i2 and lasota in commercial and local chickens in Plateau State Nigeria
U Musa, PA Abdu, UM Mera, PE Emmenna, MS Ahmed
Nigerian Veterinary Journal , 2010,
Abstract: Vaccination trials and comparative immunogenicity study using Newcastle disease vaccine strain I2 (NDVI2) and NDV La Sota administered to commercial and local chickens through intraocular (i/o), intramuscular (i/m), drinking water (dw), untreated sorghum, parboiled sorghum, sorghum coated with gum Arabic or commercial chick mash feed as vaccine carriers was conducted. Newcastle disease vaccine strain I2 and NDV La Sota vaccines provided protection to commercial and local chickens vaccinated through i/o, i/m or dw. No significant difference (P.0.05) was observed in the antibody titre of commercial or local chickens vaccinated with either NDVI2 or NDV La Sota vaccines administered via commercial feed, parboiled sorghum, parboiled sorghum coated with gum Arabic and untreated sorghum. NDVI2 or NDV La Sota vaccines administered through commercial feed, parboiled sorghum, parboiled sorghum coated with gum Arabic and untreated sorghum gave no or limited protection (0-22%) to the birds when challenged with a local strain of velogenic viscerotropic Newcastle disease virus Kudu 113 strain. It was concluded that the vaccine carriers used in this study were not suitable for delivery of NDVI2 or NDVS La Sota vaccines to local or commercial chickens.
Experimental Study of H120 Vaccination Efficacy on Respiratory Tract in Broiler Chickens  [cached]
Yousef Doustar,Adel Feizi,Mehrdad Nazeri,Amirreza Ebadi
Current Research Journal of Biological Sciences , 2012,
Abstract: Infectious bronchitis is an acute viral disease with high contagious and mortality among chicks. Apparent symptoms of Infectious bronchitis in chicks are tracheal respiration and nasal discharges. The objective of this study was to assessment H120 vaccination efficacy on respiratory tract in Broiler Chickens. In this study, 60 Ross 308 commercial strain one-day old chicks free of mycoplasma galisepticum and mycoplasma synoviae were divided into two, control and treatment, groups and three sub groups of 10 chicks. The treatment sub groups were vaccinated against bronchitis on the first day. All of chicks were placed in the hatchery and 20 mL of 10% formalin. H120 lyophilized vaccine of massachoset strain prepared in Razi institute was used for vaccination. The results of the present study show that spraying form of vaccination has a high reaction. So it is recommended that this kind of vaccination is used carefully. In the present study the aftermath of H120 vaccine spray was observed and recorded during Histopathological studies.
Kinases of two strains of Mycoplasma hyopneumoniae and a strain of Mycoplasma synoviae: an overview
Bail?o, Alexandre Melo;Parente, Juliana Alves;Pereira, Maristela;Soares, Célia Maria de Almeida;
Genetics and Molecular Biology , 2007, DOI: 10.1590/S1415-47572007000200008
Abstract: mycoplasma synoviae and mycoplasma hyopneumoniae are wall-less eubacteria belonging to the class of mollicutes. these prokaryotes have a reduced genome size and reduced biosynthetic machinery. they cause great losses in animal production. m. synoviae is responsible for an upper respiratory tract disease of chickens and turkeys. m. hyopneumoniae is the causative agent of enzootic pneumonia in pigs. the complete genomes of these organisms showed 17 orfs encoding kinases in m. synoviae and 15 in each of the m. hyopneumoniae strain. four kinase genes were restricted to the avian pathogen while three were specific to the pig pathogen when compared to each other. all deduced kinases found in the non pathogenic strain (j[atcc25934]) were also found in the pathogenic m. hyopneumoniae strain. the enzymes were classified in nine families composing five fold groups.
Involvement of Mycoplasma synoviae in Respiratory Distress Cases of Broilers  [PDF]
S. Ehtisham-ul-Haque*, S. U. Rahman, M. Siddique and A. S. Qureshi1
Pakistan Veterinary Journal , 2011,
Abstract: Mycoplasma synoviae (MS) is an important pathogen of poultry worldwide, causing respiratory tract infection and infectious synovitis in chickens and turkeys. The study was designed to detect M. synoviae through serology, culture isolation and polymerase chain reaction (PCR) assay to document the involvement of MS infection in respiratory distress cases of broiler birds. The validated PCR assay amplifying the conserved gene region of 16SrRNA gene was applied for the detection of M. synoviae from culture as well as in clinical samples. The results indicated that 04 out of total 17 commercial broiler flocks showing respiratory distress signs were found positive with M. synoviae infection indicating 76.57% sero-positivity as, determined with rapid serum agglutination (RSA) test. Out of 85 clinical specimens (collected from sero-positive birds); M. synoviae culture isolation was successfully attained in 36 (42.35%) samples. Whereas, PCR test has detected 84 (98.82%) positive cases. The prevalence of MS in broiler birds was observed maximum as measured through PCR. It is suggested that the true prevalence of MS may best be reflected by combining RSA and PCR test findings.
Response of chickens to oral vaccination with Newcastle disease virus vaccine strain I2 coated on maize offal
BC Echeonwu, MB Ngele, GON Echeonwu, TM Joannis, EM Onovoh, G Paul
African Journal of Biotechnology , 2008,
Abstract: Thermostable Newcastle disease (ND) vaccine virus strain I2 was investigated for its efficacy as foodborne vaccine, using maize offal as the vehicle. Immune response to vaccination and resistance to challenge were assessed by standard methods. Results showed that following primary vaccination, 40 (64.5%) out of the 62 birds produced detectable haemagglutination inhibiting (HI) antibody, but only 4 (6.5%) produced HI (log2) antibody titre 3.0 regarded as protective with a geometric mean titre (GMT) of 3.1. After a booster dose, 49 (79.0%) seroconverted and 20 (32.3%) had HI (log2) titres 3.0 with GMT of 4.9. When challenged all vaccinated birds survived while all control (unvaccinated) birds died. Prechallenge HI antibody titre of 50 vaccinated birds selected for challenge showed that 13 (26.0%) had titres 3.0 and GMT = 4.5, while post-challenge, 31 (62.0%) had HI (log2) 3.0 with GMT of 7.2. Using Student t test analysis of significance, the birds were observed to show 70% HI antibody production at a P 0.3 and 3 degree of freedom (df), and 70% secondary immune response on challenge at 4df. It is therefore concluded that the vaccine could be effective for protection of village chickens as food-borne vaccine provided the carrier foods are adequately treated.
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