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      Occurrence of Mycoplasma gallisepticum and avian metapneumovirus in commercial broiler flocks from the Southeast and Midwest regions of Brazil

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          Abstract

          ABSTRACT Respiratory tract infections in poultry have become serious problems with negative consequences for poultry production. Among these diseases, mycoplasmosis and avian pneumoviruses stand out, which, despite being relevant in poultry health, have not been systematically investigated in Brazil, especially regarding the interaction between these agents or the occurrence of co-infection in broiler chickens. The present study investigated the occurrence of infection by Mycoplasma gallisepticum (MG) and subtypes A and B of avian metapneumovirus (aMPV) in broiler chickens from commercial poultry flocks kept on farms with high biosecurity standards and located in the Southeast and Midwest regions of Brazil. Polymerase chain reaction (PCR) and reverse transcription nested-PCR techniques were applied to detect and/or identify, respectively, MG and aMPV in samples of nasal and tracheal swabs collected between 2017 and 2018, from 87 batches of broiler chickens from 15 commercial production farms that showed respiratory clinical signs. Two out of 87 batches sampled (2.3%) were positive for MG, while none of them were positive for aMPV. The low or no incidence of these pathogens can be explained by the adoption of increasingly effective health control measures for these agents on farms with high biosecurity standards. In addition, there are clues that other bacterial and viral infectious agents may be involved in the etiology of respiratory problems of these broiler chickens, which showed clinical signs of respiratory diseases upon sample collection.

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          Most cited references28

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          Control of avian mycoplasma infections in commercial poultry.

          S Kleven (2008)
          Control of pathogenic avian mycoplasmas can consist of one of three general approaches: Maintaining flocks free of infection, medication, or vaccination. Maintaining flocks free of pathogenic mycoplasmas consists of maintaining replacements from mycoplasma-free sources in a single-age, all-in all-out management system. Good biosecurity and an effective monitoring system are necessary aspects of this program. Medication can be very useful in preventing clinical signs and lesions, as well as economic losses, but cannot be used to eliminate infection from a flock and is therefore not a satisfactory long-term solution. Vaccination against Mycoplasma gallisepticum (MG) or M. synoviae (MS) can be a useful long-term solution in situations where maintaining flocks free of infection is not feasible, especially on multi-age commercial egg production sites.
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            Extensive sequence variation in the attachment (G) protein gene of avian pneumovirus: evidence for two distinct subgroups.

            The putative attachment protein of the avian pneumovirus that causes turkey rhinotracheitis is, by analogy with mammalian pneumoviruses, expected to be the major antigenic determinant. We report the nucleotide sequence of the attachment (G) protein genes of five different continental European isolates and compare them with the previously published sequence of the G gene for the focal variant of a U.K. isolate. The nucleotide sequences and the predicted amino acid sequences indicate that there are at least two distinct subgroups, similar to the grouping described for human respiratory syncytial (RS) virus. The U.K. and French isolates form one group and the isolates from Spain, Italy and Hungary form a second. The two subgroups can be easily distinguished on the basis of restriction enzyme digestion of PCR-generated products representing the full-length gene. Within the subgroups the predicted G proteins were highly conserved (98.5 to 99.7% amino acid identity) compared to the levels of identity of RS virus G proteins in the same subgroup (80 to 95%). Between the avian pneumovirus subgroups described here there was an unexpected degree of divergence, the average amino acid identity between members of the two groups being only 38%. This compares with the 53% conservation seen between members of the RS virus subgroups A and B. Comparison of the predicted amino acid sequences showed that the G proteins of members of the two avian pneumovirus subgroups had similar structural features. All proteins had an amino-terminal membrane anchor and the positions of cysteine residues were highly conserved. The potential importance of the high level of variation between the two subgroups in terms of epidemiology of the disease is discussed.
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              Evaluation and comparison of various PCR methods for detection of Mycoplasma gallisepticum infection in chickens.

              Four genetic Mycoplasma gallisepticum (MG) polymerase chain reactions (PCRs) (16s rRNA PCR, three newly developed PCR methods that target surface protein genes [mgc2, LP (nested) and gapA (nested)]) were compared for analytical specificity and sensitivity and for diagnostic sensitivity (Se) and specificity of detection from tracheal swabs. The licensed MG DNA Test Kit Flock Chek test (IDEXX, Laboratories, Inc., Westbrook, ME) was as well evaluated for the diagnostic specificity and sensitivity of detection from tracheal swabs. Analytical specificity was evaluated for the four generic PCR methods using a panel of DNA samples from microorganisms that may be isolated from the trachea of commercial poultry and other fowl. PCR methods mgc2, nLP, and ngapA only amplified DNA from MG, whereas 16S rRNA PCR amplified DNA from MG and Mycoplasma imitans. The analytical sensitivity of the four generic PCR methods expressed in color-changing units (CCU)/amplification reaction was estimated for each PCR method and ranged from 4 to 400 CCU/reaction; the sensitivities of single PCR methods 16S rRNA and mgc2 were estimated at 40 CCU/reaction, the nLP at 400 CCU/reaction, and the ngapA at 4 CCU/reaction. The diagnostic sensitivity and specificity of MG detection from tracheal swab pools, as compared to isolation from choanal cleft swabs, was evaluated for the five PCR methods using three groups of birds exposed to vaccine strains ts-11 and 6/85 and to challenge strain R. All PCR methods were able to detect the vaccine strains and the challenge strain R directly from tracheal swabs, indicating that PCR primers from the different methods amplified divergent MG strains. Isolation and PCR results correlated satisfactorily among the three experimentally infected groups, with agreement values (k) ranging from 0.52 to 1.00. The ngapA, IDEXX, and mgc2 PCRs showed the best sensitivity (Se) ratios for detection of M. gallisepticum strains as compared to isolation. Compared to the ngapA and IDEXX PCR methods, the mgc2 PCR has a faster turnaround time, since this test consists of a single amplification reaction and the amplification product is detected by gel electrophoresis. Therefore, among the PCR methods evaluated in this study, the mgc2 PCR is the method of choice to further validate in the field.
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                Author and article information

                Journal
                aib
                Arquivos do Instituto Biológico
                Arq. Inst. Biol.
                Instituto Biológico (São Paulo, SP, Brazil )
                0020-3653
                1808-1657
                2024
                : 91
                : e00222022
                Affiliations
                [01] Jaboticabal orgnameUniversidade Estadual Paulista orgdiv1Faculdade de Ciências Agrárias e Veterinárias orgdiv2Departamento de Patologia, Reprodução e Saúde Única Brazil
                Article
                S1808-16572024000100700 S1808-1657(24)09100000700
                10.1590/1808-1657000222022
                75d5102b-5e2d-4844-895d-c4ae874b03b8

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 15 March 2024
                : 28 October 2022
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 28, Pages: 0
                Product

                SciELO Brazil

                Categories
                SCIENTIFIC ARTICLE / Animal Epidemiology

                broiler production,PCR,respiratory diseases,RT-Nested-PCR

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