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Mycoplasma hyopneumoniae resides intracellularly within porcine epithelial cells.

ABSTRACT: Enzootic pneumonia incurs major economic losses to pork production globally. The primary pathogen and causative agent, Mycoplasma hyopneumoniae, colonises ciliated epithelium and disrupts mucociliary function predisposing the upper respiratory tract to secondary pathogens. Alleviation of disease is reliant on antibiotics, vaccination, and sound animal husbandry, but none are effective at eliminating M. hyopneumoniae from large production systems. Sustainable pork production systems strive to lower reliance on antibiotics but lack of a detailed understanding of the pathobiology of M. hyopneumoniae has curtailed efforts to develop effective mitigation strategies. M. hyopneumoniae is considered an extracellular pathogen. Here we show that M. hyopneumoniae associates with integrin ?1 on the surface of epithelial cells via interactions with surface-bound fibronectin and initiates signalling events that stimulate pathogen uptake into clathrin-coated vesicles (CCVs) and caveosomes. These early events allow M. hyopneumoniae to exploit an intracellular lifestyle by commandeering the endosomal pathway. Specifically, we show: (i) using a modified gentamicin protection assay that approximately 8% of M. hyopneumoniae cells reside intracellularly; (ii) integrin ?1 expression specifically co-localises with the deposition of fibronectin precisely where M. hyopneumoniae cells assemble extracellularly; (iii) anti-integrin ?1 antibodies block entry of M. hyopneumoniae into porcine cells; and (iv) M. hyopneumoniae survives phagolysosomal fusion, and resides within recycling endosomes that are trafficked to the cell membrane. Our data creates a paradigm shift by challenging the long-held view that M. hyopneumoniae is a strict extracellular pathogen and calls for in vivo studies to determine if M. hyopneumoniae can traffic to extrapulmonary sites in commercially-reared pigs.

SUBMITTER: Raymond BBA

PROVIDER: S-EPMC6283846 | biostudies-other | 2018 Dec

REPOSITORIES: biostudies-other

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Mycoplasma hyopneumoniae resides intracellularly within porcine epithelial cells.

Raymond B B A BBA Turnbull L L Jenkins C C Madhkoor R R Schleicher I I Uphoff C C CC Whitchurch C B CB Rohde M M Djordjevic S P SP

Scientific reports 20181206 1

Enzootic pneumonia incurs major economic losses to pork production globally. The primary pathogen and causative agent, Mycoplasma hyopneumoniae, colonises ciliated epithelium and disrupts mucociliary function predisposing the upper respiratory tract to secondary pathogens. Alleviation of disease is reliant on antibiotics, vaccination, and sound animal husbandry, but none are effective at eliminating M. hyopneumoniae from large production systems. Sustainable pork production systems strive to low ...[more]

PMID: 30523267

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Project description:BackgroundGenotypic variability in M. hyopneumoniae has been reported within and among herds. However, information regarding VNTR types within single lung lobes is lacking. The objective of his study was to analyse M. hyopneumoniae infections and their association with VNTR types and lung lesions at the lobe level. Lungs from 300 pigs from 10 farms experiencing an enzootic pneumonia outbreak were collected and scored. M. hyopneumoniae was detected by real-time PCR and genotyped by MLVA assay in all samples.ResultsThe results showed genotypic variability within single pigs and among lung lobes. At the lobe level, infection with one VNTR type (SN infection) was dominant. Lobes with lesion scores > 0 were associated with positive results for real-time PCR. At the lobe level, no relationship was observed between infections with more than one genotype (MX infections) and the proportion of Mycoplasma-like lesions. Lesion-free lobes presented a higher proportion of MX infections than lobes scored > 0. M. hyopneumoniae was detected more frequently in the right lobe of the lung (p < 0.05), with a similar distribution within lobes for SN and MX infections. The anatomic conformation of swine lungs led to a higher prevalence of infections in the right lobe. However, this study showed that this condition did not affect the distribution of infections with multiple VNTR types. Nevertheless, careful consideration of sample selection should be practised for M. hyopneumoniae genotype analyses, including lung lobes with no visible lesions.ConclusionThe results did not show a significant association between the number of detected genotypes and the severity of the lesions at the lung lobe level, but revealed the unexpected detection of M. hyopneumoniae genotypes in lesion-free lobes. These results imply that a representative sampling of all lobes may lead to an accurate identification of the VNTR-type distribution. Further studies including factors that can affect pathogenetic evolution of this bacterium could shed light on the complexity of the relationship between genotypes and the lung lesions magnitude.

Project description:BackgroundA discrete time, stochastic, compartmental model simulating the spread of Mycoplasma hyopneumoniae within a batch of industrially raised pigs was developed to understand infection dynamics and to assess the impact of a range of husbandry practices. A 'disease severity' index was calculated based on the ratio between the cumulative numbers of acutely and chronically diseased and infectious pigs per day in each age category, divided by the length of time that pigs spent in this age category. This is equal to the number of pigs per day, either acutely or chronically infectious and diseased, divided by the number of all pigs per all days in the model. The impact of risk and protective factors at batch level was examined by adjusting 'acclimatisation of gilts', 'length of suckling period', 'vaccination of suckling pigs against M. hyopneumoniae', 'contact between fattening pigs of different age during restocking of compartments' and 'co-infections in fattening pigs'.ResultsThe highest 'disease severity' was predicted, when gilts do not have contact with live animals during their acclimatisation, suckling period is 28 days, no vaccine is applied, fatteners have contact with pigs of other ages and are suffering from co-infections. Pigs in this scenario become diseased/infectious for 26.1 % of their lifetime. Logistic regression showed that vaccination of suckling pigs was influential for 'disease severity' in growers and finishers, but not in suckling and nursery pigs. Lack of contact between gilts and other live pigs during the acclimatisation significantly influenced the 'disease severity' in suckling pigs but had less impact in growing and finishing pigs. The length of the suckling period equally affected the severity of the disease in all age groups with the strongest association in nursery pigs. The contact between fatteners of different groups influenced the course of infection among finishers, but not among other pigs. Finally, presence of co-infections was relevant in growers and finishers, but not in younger pigs.ConclusionThe developed model allows comparison of different prevention programmes and strategies for controlling transmission of M. hyopneumoniae.

Dataset Information

Mycoplasma hyopneumoniae resides intracellularly within porcine epithelial cells.

Publications

Mycoplasma hyopneumoniae resides intracellularly within porcine epithelial cells.

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