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Detection of Mycoplasma synoviae Infection in Broiler Breeder Farms of Morocco Using Serological Assays and Real Time PCR

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Journal of Life Sciences 2014年第10期8卷 815-821页

作者： Nassik Saadia Aboukhalid Rachid Azzam Falak rahmatallah naoufal Lahlou-Amine Idriss Fassi-FihriOuafaa El Houadfi Mohammed Avian Pathology Unit Department of Pathology and Veterinary Public Health Agronomy and Veterinary Institute Hassan IIRabat 10000 Morocco Immunology and Biochemistry Laboratory Faculty of Sciences Mohamed V-Agdal University Rabat 10000 Morocco University Mohammed V Souissi Mohammed V Military Teaching Hospital Research and Biosafety Level 3 Laboratory Rabat10000 Morocco Microbiology Immunology and Infectious Diseases Unit Department of Pathology and Veterinary Public Health Agronomy andVeterinary Institute Hassan II Rabat 10000 Morocco

Mycoplasma synoviae （MS） is an economically important pathogenic agent of chickens, causing air sacculitis and synovitis. The diagnosis and monitoring of M. synoviae infection is usually made using serological assays, while confirmative diagnosis is made by isolation and identifcation of the organism, because of the cross reaction between M. gallisepticum and M. synoviae. This study was conducted from 2011 to 2013 during which l l broiler breeder flocks were sampled. These farms were located in all regions of morocco, The sampling was conducted as follows： Farms were visited on day one old chicks ＂day of importation from Europe＂. 20 to 60 ＂chicks＂ were randomly sampled. At the age of 8, 16, 32 and 56 weeks, 60 blood samples and 60 tracheal swabs were collected at each sampling. The serological screening was performed using Rapid Slide Agglutination （RSA） according the OIE protocol and Indirect EL1SA （IDEXX） according the manufacturer＇s instructions. The molecular diagnosis was performed using a commercial kit of a duplex real time PCR （Life Biotechnology）. The results revealed that one day old chicks were negative to MS by RSA and PCR, however they have a variable stock of maternal antibodies （Ig-Y） detected by iELISA. The seroprevalence found by RSA is variable and increase with the age （Sth week： 55%, 16th week： 91%, 32tb and 56th week： 100%）, the same profile was traced by PCR （Sth week： 36%, 16th week： 64%, 32th week： 82%, 56th week： 100%）, however, all farms were positive by iELISA, from the first day to 56th weeks. These results show that MS infections are very frequent and very widespread among poultry breeder flocks, and showed a perfect agreement between serological tests and Real time PCR starting from 32th week of age.

关键词： Mycoplasma synoviae Rapid Slide Agglutination iEL1SA PCR broiler breeders flocks

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Diagnosis of Clinical Cases of Infectious Bursal Disease Using a Modified Rapid Taq Man-MGB Real-Time RT-PCR Assay

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Journal of Agricultural Science and Technology(A) 2018年第4期8卷 230-238页

作者： Maryame Cheggag Khalil Zro Ghizlane Sebbar naoufal rahmatallah Mohamed Mouahid Mohammed EL Houadfi Faouzi Kichou Unit of Histology and Pathological Anatomy Department of Veterinary Pathology and Public HealthInstitute of Agronomy and Veterinary Medicine Hassan IIB.P.6202RabatMorocco Society of Veterinary Pharmaceutical and Biological Productions(Biopharma) B.P.4569RabatMorocco Veterinary Clinic Temara Morocco

Infectious bursal disease(IBD)is an important contagious viral infection of immune system of *** infection possesses a permanent threat to the profitability of poultry industry *** aim of this work was to modify the Taq Man-MGB real-time reverse transcription-polymerase chain reaction(rRT-PCR)in one step involving two fluorogenic Taq Man labeled probe and using this protocol for detection of infectious bursal disease virus(IBDV)collected from suspected cases distributed in different regions of the country during the period *** intralaboratory validation of modified method was realized for specificity,linearity,repeatability,sensitivity and *** allowed reducing the test running time by six *** method was applied on 102 pools of bursa of fabricius(BF)samples collected from affected broiler farms suspected to be infected by *** showing macroscopic lesions including muscle petechial hemorrhages,hypertrophy and hemorrhage of BF,were subjected to molecular analysis using modified protocol“Taq Man-MGB rRT-PCR”.The validation satisfied all criteria and the assay developed could be a useful tool for a very rapid diagnosis of IBDV and permit to detect and to discriminate in one-step very virulent(vv)from non-vv(classic and variant)IBDV *** of 84 IBDV positive samples,a prevalence of 39%for vv strains and 61%for classical strains was *** results indicate that despite the vaccination against IBDV,the vv form of this pathologie continues to cause serious problems for Moroccan broiler *** obtained results indicate the successfully detection of IBDV and differentiated all vvIBDV strains from non-vvIBDV strains;Avian infectious agent RNA viruses tested are negative,demonstrating great specificity of the *** results obtained indicate that this method is suitable as a routine laboratory test for the rapid detection and differentiation of IBDV strains in samples of avian origin.

关键词： IBDV one-step rRT-PCR Taq Man-MGB macroscopic lesions

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