Mycoplasma gallisepticum variable lipoprotein hemagglutin (vlhA) proteins are crucial for immune evasion from the host cells, permitting the persistence and survival of the pathogen. However, the exact molecular mechanism behind the immune evasion function is still not clear. In silico physiochemical analysis, domain analysis, subcellular localization, and homology modeling studies have been carried out to predict the structural and functional properties of these proteins. The outcomes of this study provide significant preliminary data for understanding the immune evasion by vlhA proteins. In this study, we have reported the primary, secondary, and tertiary structural characteristics and subcellular localization, presence of the transmembrane helix and signal peptide, and functional characteristics of vlhA proteins from M. gallisepticum strain R low. The results show variation between the structural and functional components of the proteins, signifying the role and diverse molecular mechanisms in functioning of vlhA proteins in host immune evasion. Moreover the 3D structure predicted in this study will pave a way for understanding vlhA protein function and its interaction with other molecules to undergo immune evasion. This study forms the basis for future experimental studies improving our understanding in the molecular mechanisms used by vlhA proteins.

Respiratory tract infections are of serious concern to the poultry industry. The present study was aimed to delineate the extent of respiratory avian mycoplasmosis associated bacterial and viral concurrent infections in the poultry flocks. A total of 146 poultry flocks of Haryana and Rajasthan, India, suspected for chronic respiratory disease (CRD) were screened for avian mycoplasmas, Newcastle disease virus (NDV), infectious bronchitis virus (IBV), and avian pathogenic Escherichia coli (APEC) by conventional polymerase chain reaction (PCR) assays. A total of 49.31% (72/146) flocks were found positive for Mycoplasma infection. Of the Mycoplasma-positive flocks, 80.55% (58/72) represented pathogenic avian mycoplasmas (MG and/or MS), while 19.44% (14/72) flocks were positive for commensal avian mycoplasmas (other than MG and MS). A correlation was deduced between avian mycoplasmosis and bacterial and/or viral co-infections. The results revealed that 17.24% (10/58) flocks had only avian mycoplasmosis infection. However, in the remaining flocks, the avian mycoplasmosis was associated either with APEC infection [17.24% (10/58)], IBV infection [43.10% (25/58)], or both APEC and IBV infections [22.41% (13/58)], respectively. Further epidemiological studies on respiratory avian mycoplasmosis associated concurrent infections with other pathogens are recommended to assess circulating strains, risk factors, and economic losses.

Worldwide, Mycoplasma gallisepticum (MG) and M. synoviae (MS) are the main agents responsible for chronic respiratory disease in poultry. Therefore, we conducted a systematic review and meta-analysis to estimate their occurrence. We searched electronic databases to find peer-reviewed publications reporting the molecular detection of MG and MS in poultry and used meta-analysis to estimate their pooled global occurrence (combined flock and individual), aggregating results at the regional and national levels. We performed a subgroup meta-analysis for subpopulations (broilers, layers, breeders and diverse poultry including turkeys, ducks and ostriches) and used meta-regression with categorical modifiers. We retrieved 2294 publications from six electronic databases and included 85 publications from 33 countries that reported 62 studies with 22,162 samples for MG and 48 studies with 26,413 samples for MS. The pooled global occurrence was 38.4% (95% CI: 23.5-54.5) for MS and 27.0% (20.4-34.2) for MG. Among regions, Europe and Central Asia had the lowest occurrence for both pathogens, while MG and MS were highly prevalent in South Asia and sub-Saharan Africa, respectively. At the national level, MG occurrence was higher in Algeria, Saudi Arabia and Sudan, whereas China, Egypt and Ethiopia reported higher values of MS. Among the poultry subpopulations, MS and MG were more prevalent in the breeders and layers (62.6% and 31.2%, respectively) than in diverse poultry. The year of publication, the sample size and the level of ambient air pollution (measured indirectly by PM2.5) were associated with the occurrence of both mycoplasmas. Our study revealed high and heterogeneous occurrence values of MG and MS and justifies the need for early detection and improved control measures to reduce the spread of these pathogens.

Avian mycoplasmosis mainly caused by Mycoplasma gallisepticum and M. synoviae is an economically important disease of poultry industry. It causes huge economic losses in terms of decrease in weight gain, feed conversion efficiency, egg production, hatchability; increase in embryo mortality, carcass condemnation, prophylaxis and treatment cost in broiler, layer and breeder flocks. The disease is caused by four major pathogenic mycoplasmas viz., M. gallisepticum (MG), M. synoviae (MS), M. meleagradis (MM) and M. iowae (MI). The MG and MS are World Organization for Animal Health listed respiratory pathogens. MG causes chronic respiratory disease in chicken and infectious sinusitis in turkey; however, MS causes synovitis and airsacculitis in birds. The infection is transmitted both horizontally and vertically. Prevention and control measures of avian mycoplasmosis mainly comprises of biosecurity, treatment and vaccination. For vaccination of birds, inactivated bacterins, live attenuated and/or recombinant live poxvirus vaccines are commercially available against MG and MS infection. The present systematic review summarizes the different epidemiological studies carried out on MG and MS infection in poultry in different geographical locations of India and abroad over the last decade (2010-2020), economic impact, diagnosis and prevention and control.

Avian mycoplasmosis (Mycoplasma gallisepticum, Mycoplasma meleagridis) has been assessed according to the criteria of the Animal Health Law (AHL), in particular criteria of Article 7 on disease profile and impacts, Article 5 on the eligibility of avian mycoplasmosis to be listed, Article 9 for the categorisation of avian mycoplasmosis according to disease prevention and control rules as in Annex IV and Article 8 on the list of animal species related to avian mycoplasmosis. The assessment has been performed following a methodology composed of information collection and compilation, expert judgement on each criterion at individual and, if no consensus was reached before, also at collective level. The output is composed of the categorical answer, and for the questions where no consensus was reached, the different supporting views are reported. Details on the methodology used for this assessment are explained in a separate opinion. According to the assessment performed, avian mycoplasmosis can be considered eligible to be listed for Union intervention as laid down in Article 5(3) of the AHL. The disease would comply with the criteria as in Sections 4 and 5 of Annex IV of the AHL, for the application of the disease prevention and control rules referred to in points (d) and (e) of Article 9(1). The assessment here performed on compliance with the criteria as in Section 3 of Annex IV referred to in point (c) of Article 9(1) is inconclusive. The animal species to be listed for avian mycoplasmosis according to Article 8(3) criteria are mainly domestic and wild birds of the order Galliformes, and also Passeriformes for M. gallisepticum.

Mycoplasma gallisepticum causes respiratory diseases and reproduction disorders in turkeys and chickens. The infection has considerable economic impact due to reduced meat and egg production. Because elimination programmes are not feasible in a large number of poultry farms, vaccination remains the only effective measure of disease control. Differentiating vaccine strains from field isolates is necessary in the control of vaccination programmes and diagnostics. The aim of this study was to develop a polymerase chain reaction based mismatch amplification mutation assay (MAMA) for the discrimination of K vaccine strain (K 5831, Vaxxinova Japan K.K.). After determining the whole genome sequence of the K strain, primers were designed to detect seven different vaccine-specific single nucleotide polymorphisms. After evaluating preliminary results, the MAMA-K-fruA test detecting a single guanine-adenine substitution within the fruA gene (G88A) was found to be the most applicable assay to distinguish the K vaccine strain from field isolates. The detected K strain-specific single nucleotide polymorphism showed genetic stability after serial passage in vitro, but this stability test should still be evaluated in vivo as well, investigating a large number of K strain re-isolates. The MAMA-K-fruA assay was tested on a total of 280 culture and field samples. The designed assay had 102 and 103 template copy number/µl sensitivity in melt-curve analysis based and agarose-gel based assays, respectively, and showed no cross reaction with other avian Mycoplasma species. The new MAMA provides a time- and cost-effective molecular tool for the control of vaccination programmes and for diagnostics.

BACKGROUND: In Southeast Asian countries, including Myanmar, poultry farming is a major industry. In order to manage and maintain stable productivity, it is important to establish policies for biosecurity. Infectious respiratory diseases are a major threat to poultry farming. Avian influenza and Newcastle disease have been reported in Myanmar, but no scientific information is available for other respiratory pathogens, such as mycoplasmas and infectious bronchitis virus (IBV). Identifying the genotypes and serotypes of IBVs is especially important to inform vaccination programs. In this study, we detected Mycoplasma gallisepticum (MG), M. synoviae (MS), and IBV in several poultry farms in Myanmar.
RESULTS: Samples were collected from 20 farms in three major poultry farming areas in Myanmar, and MG, MS, and IBV were detected on two, four, and eight farms, respectively, by polymerase chain reaction. Phylogenetic analysis revealed that the observed MG and MS isolates were not identical to vaccine strains. Three different genotypes of IBV were detected, but none was an unknown variant.
CONCLUSIONS: Mycoplasmas and IBV were detected on poultry farms in Myanmar. Periodic surveillance is required to establish the distribution of each pathogen, and to institute better vaccine protocols.

OBJECTIVE: Avian mycoplasmosis, particularly Mycoplasma gallisepticum (MG) is one of the infectious diseases associated with economic losses in Egyptian poultry industry. Thus, this study was aimed to determine the prevalence, serological identification, molecular characterization, sequencing and minimum inhibitory concentration of M. gallisepticum isolated from diseased broilers in Egypt.
METHODS: A total of 351 samples (227 tissue samples \"tracheas and air sacs\" and 124 tracheal swabs) and 71 sera were collected from diseased broilers. The conventional (isolation and biochemical) and molecular methods (PCR) were performed for detection of M. gallisepticum and virulence-associated gene (mgc2). The serum plate agglutination (SPA) test and enzyme-linked immunosorbent assay (ELISA) were applied on sera for determination of the presence of antibodies against M. gallisepticum. The minimal inhibitory concentration test (MIC) was used to determine the sensitivity of two sequenced M. gallisepticum strains to anti-mycoplasma agents.
RESULTS: The total recovery rate of Mycoplasma from 351 samples from broilers was 45.29% (159) in which M. gallisepticum showed a prevalence of 62.89% (100/159). Serological identification of M. gallisepticum in 71 collected sera using SPA and ELISA were 54.9 and 40.8% with the highest geometric mean titer of ELISA for M. gallisepticum (699.08 and 495.92). Molecular characterization of Mycoplasma using PCR showed that 50% (3/6) of tested isolates were identified as M. gallisepticum based on 16SrRNA. Also, the mgc2 gene was detected in 50% (3/6) M. gallisepticum isolates. Two positive PCR mgc2 specific genes of M. gallisepticum isolates were subjected to gene target sequencing (GTS) to verify that these two isolates were M. gallisepticum. The minimal inhibitory concentration test (MIC) was applied to determine the sensitivity of these two sequenced M. gallisepticum strains to anti-mycoplasma agents. The first M. gallisepticum isolate was sensitive to tilmicosin, tiamulin and spiramycin. The second M. gallisepticum isolate showed sensitivity to tiamulin, spiramycin and tilmicosin.
CONCLUSIONS: These results summarized the necessity of monitoring the Egyptian poultry farms for avian mycoplasmosis. Also, further studies are required for controlling of mycoplasma in all stages of the poultry industry production chain to avoid different losses in Egypt.

Mycoplasma gallisepticum and Mycoplasma synoviae are the causative agents of avian mycoplasmosis in commercial poultry. Among the available tools, polymerase chain reaction (PCR) and culture are confirmatory tools for the diagnosis of mycoplasmosis after the initial serological screening of suspected birds. Overall, 181 samples were analyzed, 152 (84%) and 103 (57%) of which were found positive by PCR and culture, respectively. Further, 54 (92%) broiler samples were found positive for general avian mycoplasma. Among the total positive samples, MS positivity was as high as 72 (47%) by PCR, while it was 45 (44%) by culture. MG positivity was 23% and 25% in PCR- and culture-positive samples. MG grows more easily compared to MS. The agreement value between the tests was 67%. Overall, flock wise prevalence was not much varied. The prevalence of mycoplasmosis was higher during winter. Our study confirmed that PCR is the most sensitive and reliable tool for the diagnosis of avian mycoplasmosis in field samples.