Evaluation of a PCR multiplex for detection and differentiation of Mycoplasma synoviae, M. gallisepticum, and M. gallisepticum strain F-vaccine

Mettifogo, Elena; Buzinhani, Melissa; Buim, M. R.; Timenetsky, Jorge; Ferreira, Antônio José Piantino

doi:10.1590/s0100-736x2015000100004

Cited by 8 publications

(7 citation statements)

References 12 publications

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“…PCR is also chosen due to the fact that it is a specific, sensitive method capable of detecting and amplifying low-quality DNA (Saiki et al 1985, Innis & Gelfand, 1990, which assists in farm monitoring programs and the differentiation of the field and vaccine strains of MG (Nascimento et al 1993, Mettifogo et al 2002. In the present study, the PCR results for MG differentiated the MG-F vaccine in samples from the commercial egg-laying farm with a history of vaccination.…”

Section: Discussionmentioning

confidence: 87%

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Occurrence of Mycoplasma synoviae on commercial poultry farms of Pernambuco, Brazil

et al. 2014

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Pesq. Vet. Bras. 34(10):953-956, outubro 2014 953 RESUMO.-[Ocorrência de Mycoplasma synoviae em granjas comerciais da avicultura de Pernambuco.] O estado de Pernambuco é o maior produtor de ovos da região Norte e Nordeste e ocupa a segunda posição na produção de frangos de corte. Os micoplasmas são importantes patógenos aviários que causam doenças respiratórias e sinovite que resultam em grandes perdas econômicas.Objetivou-se pesquisar a ocorrência de Mycoplasma gallisepticum (MG) e Mycoplasma synoviae (MS) em frangos de corte e poedeiras comerciais no Estado de Pernambuco, Brasil. Foram colhidos fragmentos de traquéia de 55 frangos de corte sadios, 35 com sinais respiratórios e de 30 poedeiras comerciais também com sinais respiratórios, provenientes de 24 granjas, cada amostra foi composta por um "pool" de cinco aves. Para detecção de Mycoplasma gallisepticum (MG) e Mycoplasma synoviae (MS) foram utilizados o exame bacteriológico, PCR e Nested-PCR. Todas as amostras apresentaram resultados negativos no exame bacteriológico. Na PCR, sete amostras foram positivas para MS e uma para MG em amostras de aves com sinais respiratórios, sendo a amostra positiva para MG confirmada como cepa vacinal MG-F. A ocorrência de MS em aves com sinais clínicos respiratórios pode indicar ausência de barreiras sanitárias adequadas em granjas de frangos de corte e de poedeira comercial, favorecendo a sua propagação. The state of Pernambuco is the largest producer of eggs in the North and Northeast of Brazil and second one in the broiler production. Mycoplasmas are important avian pathogens, which cause respiratory and joint diseases that result in large economic losses. The aim of the present study was to investigate the occurrence of Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) in broilers and commercial laying hens in the state of Pernambuco, Brazil. Tracheal fragments were analyzed from 55 healthy broilers, 35 broilers with respiratory signs and 30 commercial laying hens with respiratory signs, from 24 commercial poultry farms, each sample was composed of a pool of five birds. The bacteriological exam, PCR and nested PCR were used for the detection of Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS). All samples were negative in bacteriological isolation. In the PCR analyses, seven samples from birds with respiratory signs were positive for MS and one was positive for MG, the latter of which was confirmed as the MG-F vaccine strain. The occurrence of MS in chickens with respiratory signs may indicate inadequate sanitary management on poultry farms, favoring the propagation of mycoplasmosis. Occurrence of

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Section: Discussionmentioning

confidence: 87%

“…In Brazil, the frequency of infection by MS is greater than that by MG (Reis et al 1973, Mettifogo et al 2002, Buim 2005, Buim et al 2009). Moreover, the prevalence of MS in chickens has increased in the past twenty years (Balen & Fiorentin 1990).…”

Section: Discussionmentioning

confidence: 99%

Occurrence of Mycoplasma synoviae on commercial poultry farms of Pernambuco, Brazil

et al. 2014

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“…MG infections usually lead to chronic respiratory disease (CRD) in chickens and infectious sinusitis in turkeys. MS causes subclinical upper respiratory infections, eggshell apex abnormalities, and tenosynovitis or bursitis in chickens and turkeys [2][3][4]. CRD caused by MG is prevalent in layer, broiler, and breeder poultry flocks.…”

Section: Introductionmentioning

confidence: 99%

Detection and antibiotic resistance of Mycoplasma gallisepticum and Mycoplasma synoviae among chicken flocks in Egypt

Emam¹,

Hashem

Elhariri

et al. 2020

Vet World

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Background and Aim: Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) are the most significant pathogens of avian mycoplasmosis. This study aimed to isolate and identify MG and MS from chickens and detect the various virulence genes in the isolates. Moreover, the efficacies of different antibiotics were tested to identify suitable treatment regimens. Materials and Methods: We isolated MG and MS from 487 chicken samples of different ages located in different Governorates in Egypt using conventional isolation methods. The isolates were characterized by polymerase chain reaction (PCR) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then tested for antibiotic sensitivity by the minimum inhibitory concentration (MIC) method. Results: The prevalence of MG among the isolates was 9.85%, with the highest percentage isolated from air sacs, while the prevalence of MS among the isolates was 1.6%. Moreover, the highest levels of the prevalence of both MG and MS were during the winter and autumn sampling, while the lowest levels were in the summer and spring. Following the 16S rRNA-based detection of Mycoplasma isolates, 14 MG and 5 MS isolates were identified by different PCR-based detection methods for various virulence genes. Nine MG isolates contain the mgc2 gene, six MG isolates contain the gapA gene, and three MS isolates contain the vlhA gene. We validated a duplex PCR method for the simultaneous identification of MG and MS, based on 100% of the MG and MS isolates generating common bands at 55 and 17 kDa, respectively. The MIC method identified tiamulin and spiramycin as the antibiotics of choice for the treatment of MG and MS infections, respectively. Conclusion: For more precise diagnosis of Mycoplasma infections in chicken flocks, conventional isolation methods must be confirmed by PCR. SDS-PAGE analysis helps in epidemiological studies and vaccine preparation. The MIC method can be used to help develop therapies to control avian mycoplasmosis infections.

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“…Microbiological investigation indicated the presence of E. coli in all organ samples. Additionally, molecular testing of the liver, trachea, and lugs using an M. gallisepticum polymerase chain reaction (PCR) methodology reported by Mettifogo et al [ 6 ] revealed M. gallisepticum positivity, which was verified by sequencing (Figure not shown).…”

Section: Resultsmentioning

confidence: 86%

“…On this farm, the source(s) of M. gallisepticum infection was(were) not determined. However, the virus is considered to be transferred vertically (transovarially) from the breeder or during hatching [ 6 , 10 ]. While infected embryos may die, infected chicks may hatch and disseminate the virus throughout the flock [ 11 ].…”

Section: Discussionmentioning

confidence: 99%

Complex chronic respiratory disease concurrent with coccidiosis in broiler chickens in Malaysia: A case report

Kamaruzaman¹,

Ng²,

Hamdan³

et al. 2021

J Adv Vet Anim Res

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Objective: The case study describes the cause of an increase in mortality rates among 35-day-old broilers that developed respiratory distress and bloody diarrhea on a farm in Malaysia. Materials and Methods: The organ samples were subjected to laboratory testing and postmortem inspection. Escherichia (E.) coli and Mycoplasma (M.) gallisepticum were detected using bacterial isolation and molecular diagnostics using polymerase chain reaction. Results: Chickens with the infection had widespread fibrin buildup in several organs and hemorrhages on the duodenal mucosa. Additional histology and laboratory analysis of organ samples revealed infection with M. gallisepticum , E. coli , and enteric Eimeria spp., all of which are consistent with complex chronic respiratory disease (CCRD) associated with coccidiosis. Tylosin tartrate 20% (w/w) (2.5 gm/l) was prescribed for 1 week along with a combination of the broad-spectrum bacteriostatic drug streptomycin (25 mg/kg) and coccidiostat (2 gm/5 l). Conclusion: CCRD and coccidiosis are both infectious diseases that can infect chicken flocks, resulting in production losses and carcass quality degradation. Early disease detection and proper treatment should be provided promptly, and tight farm biosecurity should be implemented to prevent chicken mortality on the farm, as was achieved successfully.

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Evaluation of a PCR multiplex for detection and differentiation of Mycoplasma synoviae, M. gallisepticum, and M. gallisepticum strain F-vaccine

Cited by 8 publications

References 12 publications

Occurrence of Mycoplasma synoviae on commercial poultry farms of Pernambuco, Brazil

Occurrence of Mycoplasma synoviae on commercial poultry farms of Pernambuco, Brazil

Detection and antibiotic resistance of Mycoplasma gallisepticum and Mycoplasma synoviae among chicken flocks in Egypt

Complex chronic respiratory disease concurrent with coccidiosis in broiler chickens in Malaysia: A case report

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