Welkom bij het Lymeforum

Abstract

Biofilms are communities of microorganisms that are encased in polymeric matrixes and grow attached to biotic or abiotic surfaces. Despite their enhanced ability to resist antimicrobials and components of the immune system in vitro, few studies have addressed the interactions of biofilms with the host at the organ level. Although mycoplasmas have been shown to form biofilms on glass and plastic surfaces, it has not been determined whether they form biofilms on the tracheal epithelium. We developed a tracheal organ-mounting system that allowed the entire surface of the tracheal lumen to be scanned using fluorescence microscopy. We observed the biofilms formed by the murine respiratory pathogen Mycoplasma pulmonis on the epithelium of trachea in tracheal organ culture and in experimentally infected mice and found similar structure and biological characteristics as biofilms formed in vitro. This tracheal organ-mounting system can be used to study interactions between biofilms formed by respiratory pathogens and the host epithelium and to identify the factors that contribute to biofilm formation in vivo.

Introduction

Mycoplasmas are significant and chronic pathogens, of man and animals, that primarily colonize the mucosal surfaces of the respiratory and genitourinary tracts (Cassell et al., 1985). Although they lack a cell wall and would appear to be sensitive to the host's immune system, the mycoplasmas persist despite an intense inflammatory response. A key to the survival of Mycoplasma pulmonis is the V-1 variable surface antigen (Vsa protein). The length of the tandem repeat region of the Vsa protein modulates biofilm formation and the susceptibility of individual mycoplasma cells to killing by complement (Simmons & Dybvig, 2003; Simmons et al., 2004, 2007). Mycoplasmas producing a short Vsa protein containing few tandem repeats are sensitive to complement when dispersed but form complement-resistant biofilms on glass and plastic surfaces. Mycoplasmas producing a long Vsa protein containing as many as 40–60 tandem repeats do not form biofilms but are nevertheless resistant to complement.

....................

Concluding remarks

These results indicate the biofilms formed on tracheal epithelium of three different strains of mice are similar to the biofilms that are formed in vitro. The structure of the towers formed on the tracheal epithelium is similar to the structure of the towers in mycoplasma biofilms formed in vitro, with similar resistance to penetration by antibodies and dependence on a short Vsa protein. The towers are densely packed with mycoplasma cells that have a typical diameter of 500 nm. Towers with a 20-μm2 cross-section and a height of 10 μm might contain over 20 000 cells and represent a substantial reservoir of mycoplasmas that are resistant to host immunity and from which chronic infections could be maintained.

As the ability of the mycoplasmas to form biofilms in vivo may parallel their ability to form biofilms in TOC, an examination of the effects of virulence factors on biofilm formation in TOC may provide an indicator of how those factors affect virulence in the animal. Additionally, the tracheal organ-mounting system described here can be applied to biofilms formed by other bacterial species to test the validity of the current models for biofilm formation that have been developed in vitro. Rather than inferring that biofilms form on the tracheal epithelium in vivo by extrapolating from in vitro data, direct observations of biofilm formation can be made and it can be determined whether factors that contribute to in vitro spatial and temporal organization of biofilms perform similar functions ex vivo or in vivo.

Abstract

Mycoplasma pneumoniae causes acute and chronic lung infections in humans, leading to a variety of pulmonary and extrapulmonary sequelae. Of the airway complications of M. pneumoniae infection, M. pneumoniae-associated exacerbation of asthma and pediatric wheezing are emerging as significant sources of human morbidity. However, M. pneumoniae products capable of promoting allergic inflammation are unknown. Recently, we reported that M. pneumoniae produces an ADP-ribosylating and vacuolating toxin termed the community-acquired respiratory distress syndrome (CARDS) toxin. Here we report that naive mice exposed to a single dose of recombinant CARDS (rCARDS) toxin respond with a robust inflammatory response consistent with allergic disease. rCARDS toxin induced 30-fold increased expression of the Th-2 cytokines IL-4 and IL-13 and 70- to 80-fold increased expression of the Th-2 chemokines CCL17 and CCL22, corresponding to a mixed cellular inflammatory response comprised of a robust eosinophilia, accumulation of T cells and B cells, and mucus metaplasia. The inflammatory responses correlate temporally with toxin-dependent increases in airway hyperreactivity characterized by increases in airway restriction and decreases in lung compliance. Furthermore, CARDS toxin-mediated changes in lung function and histopathology are dependent on CD4(+) T cells. Altogether, the data suggest that rCARDS toxin is capable of inducing allergic-type inflammation in naive animals and may represent a causal factor in M. pneumoniae-associated asthma.

Mycoplasma species are the smallest free-living organisms. These organisms are unique among prokaryotes in that they lack a cell wall, a feature largely responsible for their biologic properties such as their lack of a reaction to Gram stain and their lack of susceptibility to many commonly prescribed antimicrobial agents, including beta-lactams. Mycoplasmal organisms are usually associated with mucosal surfaces, residing extracellularly in the respiratory and urogenital tracts. They rarely penetrate the submucosa, except in the case of immunosuppression or instrumentation, when they may invade the bloodstream and disseminate to different organs and tissues throughout the body.

Although scientists have isolated at least 17 species of Mycoplasma from humans, 4 types of organisms are responsible for most clinically significant infections that may come to the attention of practicing physicians. These species are Mycoplasma pneumoniae, Mycoplasma hominis, Mycoplasma genitalium, and Ureaplasma species. The focus of this article is infections caused by M pneumoniae; articles on Ureaplasma infections (eg, Ureaplasma Infection) and genital mycoplasmal infections contain discussions of infections caused by other mycoplasmal species.

Melkcontrolecentrum Vlaanderen en de faculteit diergeneeskunde de bacterie aan bij 7% van willekeurige tankmelkscreenings van 100 bedrijven. In 2009 was dat nog maar bij 1,5% van de geteste bedrijven.

Bij jongvee is de prevalentie van mycoplasma veel groter. Bij 37,1% van de longspoelingen voor bepaling van kalvergriep werd het afgelopen half jaar mycoplasma aangetoond. Insleep gebeurt meestal via aankoop van vee, maar ook erfbetreders kunnen een infectie binnen brengen. Binnen het bedrijf verspreidt de bacterie zich via neus-neuscontact of door het voeren van besmette melk. Mycoplasma is moeilijk met antibiotica te bestrijden.

In conclusion, while PCR and serology may be of limited value in the diagnosis of M. pneumoniae encephalitis, the detection of intrathecal antibodies to M. pneumoniae, including cross-reactive antibodies against GalC and gangliosides, may be regarded as a promising new diagnostic tool.

Abstract


In Lyme disease concurrent infections frequently occur. The clinical and pathological impact of co-infections was first recognized in the 1990th, i.e. approximately ten years after the discovery of Lyme disease. Their pathological synergism can exacerbate Lyme disease or induce similar disease manifestations. Co-infecting agents can be transmitted together with Borrelia burgdorferi by tick bite resulting in multiple infections but a fraction of co-infections occur independently of tick bite. Clinically relevant co-infections are caused by Bartonella species, Yersinia enterocolitica, Chlamydophila pneumoniae, Chlamydia trachomatis, and Mycoplasma pneumoniae. In contrast to the USA, human granulocytic anaplasmosis (HGA) and babesiosis are not of major importance in Europe. Infections caused by these pathogens in patients not infected by Borrelia burgdorferi can result in clinical symptoms similar to those occurring in Lyme disease. This applies particularly to infections caused by Bartonella henselae, Yersinia enterocolitica, and Mycoplasma pneumoniae. Chlamydia trachomatis primarily causes polyarthritis. Chlamydophila pneumoniae not only causes arthritis but also affects the nervous system and the heart, which renders the differential diagnosis difficult. The diagnosis is even more complex when co-infections occur in association with Lyme disease. Treatment recommendations are based on individual expert opinions. In antibiotic therapy, the use of third generation cephalosporins should only be considered in cases of Lyme disease. The same applies to carbapenems, which however are used occasionally in infections caused by Yersinia enterocolitica. For the remaining infections predominantly tetracyclines and macrolides are used. Quinolones are for alternative treatment, particularly gemifloxacin. For Bartonella henselae, Chlamydia trachomatis, and Chlamydophila pneumoniae the combination with rifampicin is recommended. Erythromycin is the drug of choice for Campylobacter jejuni.



INTRODUCTION

In Lyme disease, other infections, whose pathological synergism exacerbate the disease or induce similar clinical manifestations, can exist concurrently. Such concomitant infections are termed co-infections. Co-infections can be transmitted together with Borrelia burgdorferi by tick-bite, and result in multiple infection. Part of co-infections is independent of tick-bite.

The goal of this review was to summarize the more important co-infections completed with some personal experiences and with a short summary on reactive arthritis. Because of the similarity of the clinical symptoms of tularemia, Q fever, parvovirus B19 and Campylobacter jejuni infections to those of Lyme disease a short summary of these infections are also included.


RELEVANT CO-INFECTIONS IN LYME DISEASE

Co-infections can exacerbate Lyme disease through immune system modulation and are considered to be the major cause for
resistance to therapy [1-17]. The importance of co-infections in the disease process, i.e. their pathogenicity compared to Lyme disease, has not been clarified. In cases with double or multiple infections, to determine which infection predominates in the pathological process is difficult. There are substantial overlaps between the clinical symptoms caused by co-infections and Lyme disease. Consequently, an unequivocal assignment of the manifestations of the disease to existing infections might be difficult. The diagnostic difficulties of Lyme disease and co-infections always concern chronic Lyme disease (late Lyme disease, stage III). The synergic-pathological mechanism requires that co-infections are also present in chronic persistent form. Anamnestic consideration of the acute form of co-infections may be helpful to recognize their persistence in the chronic stage.


The significant co-infections in Lyme disease are caused by various Bartonella species, primarily Bartonella henselae, by Chlamydia trachomatis, Chlamydophila pneumoniae, Yersinia enterocolitica, and Mycoplasma pneumoniae

........

Mycoplasma pneumoniae Infection

The differential diagnosis between Lyme disease und Mycoplasma pneumoniae infection or the recognition of the co-infection by Mycoplasma pneumoniae is problematical because both diseases exhibit similar manifestations; this applies to the extrapulmonary manifestations of Mycoplasma pneumoniae infection: disorders of the CNS, musculoskeletal system, heart, kidney and eye.

.........

Mycoplasma pneumoniae is considered to be the most important pathogen of atypical pneumonia. However, pneumonia only occurs in approximately 3% - 10% of the cases in Mycoplasma pneumoniae infection [165]. In most cases, the infection results in a banal bronchitis [165], pharyngitis, rhinitis, earaches, and sinusitis [163].

All the extrapulmonary disease manifestations listed in Table ​99 are seldom [166-174]. In patients with arthritis, Mycoplasma pneumoniae was detected in the synovial fluid by PCR [171], which is an indication of a direct relationship to infection.

The detection of pathogen in articular effusion and many extrapulmonary manifestations of the disease document the chronic course of the disease in cases with Mycoplasma pneumoniae infection. However, precise data on the chronic course of the disease are not available in the literature. In particular, whether a chronic infection, especially with extrapulmonary disease manifestation, can persist with seronegativity is unclear. Seropositivity documents infection, but cannot serve as a proof for chronic persistent Mycoplasma pneumoniae infection.

The literature on the relationship between Mycoplasma pneumoniae infection and neurological disease manifestations is extensive. The publications primarily refer to neurological complications in pneumonia, i.e. the early phase of Mycoplasma pneumoniae infection. Neurological manifestations involve both the early phase, i.e. the point in time of existing pneumonia due to Mycoplasma pneumoniae, and later disease stages. Changes in the region of the brain stem [175, 176], myelitis [177-187], Guillain-Barré syndrome [188-193], encephalitis [185,187,194-200], meningitis [194], polyradiculopathy [178], peripheral facial paresis [201, 202], optical neuritis and hemorrhagic leukoencephalitis [190], peripheral polyneuropathy[194], cranial nerve neuritis [192], radiculitis [192] have been described.

The frequency of neurological symptoms in connection with Mycoplasma pneumoniae varies between 1‰ [203], 1% [204], and 5% [205]. The pathogen has been repeatedly detected by means of culture methods or PCR [182,193,194].


The detection of pathogen in the serum and liquor is considered to be proof that the neurological manifestations are mediated by direct infection and not by immunological responses [193]. However, the connection between Mycoplasma pneumoniae and neurological manifestations is not undisputed [203, 206].

Other extrapulmonary manifestations mentioned in the literature include hepatitis, hemolytic anemia, Schönlein-Henoch purpura, disorders of the muscular-skeletal system, of the skin and other organs [179], macula edema[194], bilateral uveitis [207], nephritis [208], arthritis, hepatitis and pericarditis [208].

The laboratory diagnostics for Mycoplasma pneumoniae include the serology, which as in most infectious diseases becomes positive after several weeks. Seroconversion is therefore significant for the chronic disease course. Seropositivity substantiates the infection, but not the disease. Whether a chronic infection can also exist in seronegative cases has not yet been scientifically clarified. The LTT for Mycoplasma pneumoniae has not yet been validated.

Detection of the pathogen, e.g. in articular effusion by PCR and culture is possible but difficult and has a low sensitivity. Consequently it is not part of the routine diagnostic procedures.
........