Interaction of Mycoplasma synoviae with chicken synovial sheath cells contributes to macrophage recruitment and inflammation.

Mycoplasma synoviae (MS) is an important avian pathogen causing considerable economic hardship in the poultry industry. A major inflammation caused by MS is synovitis that occurs in the synovial tendon sheath and joint synovium. However, the overall appearance of pathological changes in the tendon sheath and surrounding tissues caused by MS infection at the level of pathological tissue sections was poor. Studies on the role of MS and synovial sheath cells (SSCs) interaction in the development of synovitis have not been carried out. Through histopathological observation, our study found that a major MS-induced pathological change of the tendon sheath synovium was extensive scattered and focal inflammatory cell infiltration of the tendon sheath synovial layer. In vitro research experiments revealed that the CFU numbers of MS adherent and invading SSC, the levels of expression of various pattern recognition receptors, inflammatory cytokines, and chemokines coding genes, such as IL-1ß, IL-6, IL-8, CCL-20, RANTES, MIP-1ß, TLR7, and TLR15 in SSCs, and chemotaxis of macrophages were significantly increased when the multiplicity of infection (MOI) of MS to SSC were increased tenfold. The expression level of IL-12p40 in SSC was significantly higher when the MOIs of MS to SSC were increased by a factor of 100. The interaction between MS and SSC can activate macrophages, which was manifested by a significant increase in the expression of IL-1ß, IL-6, IL-8, CCL-20, RANTES, MIP-1ß, and CXCL-13. This study systematically demonstrated that the interaction of MS with chicken SSC contributes to the inflammatory response caused by the robust expression of related cytokines and macrophage chemotaxis. These findings are helpful in elucidating the molecular mechanism of MS-induced synovitis in chickens.

Spirochete antigens persist near cartilage after murine Lyme borreliosis therapy.

An enigmatic feature of Lyme disease is the slow resolution of musculoskeletal symptoms that can continue after treatment, with some patients developing an inflammatory arthritis that becomes refractory to antibiotic therapy. Using intravital microscopy and the mouse model of Lyme borreliosis, we observed that Borrelia burgdorferi antigens, but not infectious spirochetes, can remain adjacent to cartilage for extended periods after antibiotic treatment. B. burgdorferi was not recovered by culture or xenodiagnosis with ticks after antibiotic treatment of WT mice and all but one of the immunodeficient mice with heightened pathogen burden due to impaired TLR responsiveness. Amorphous GFP+ deposits were visualized by intravital microscopy in the entheses of antibiotic-treated mice infected with GFP-expressing spirochetes and on the ear cartilage surface in sites where immunofluorescence staining detected spirochete antigens. Naive mice were not infected by tissue transplants from antibiotic-treated mice even though transplants contained spirochete DNA. Tissue homogenates from antibiotic-treated mice induced IgG reactive with B. burgdorferi antigens after immunization of naive mice and stimulated TNF-α production from macrophages in vitro. This is the first direct demonstration that inflammatory B. burgdorferi components can persist near cartilaginous tissue after treatment for Lyme disease. We propose that these deposits could contribute to the development of antibiotic-refractory Lyme arthritis.

Lyme neuroborreliosis in 2 horses.

Lyme neuroborreliosis--characterized as chronic, necrosuppurative to nonsuppurative, perivascular to diffuse meningoradiculoneuritis--was diagnosed in 2 horses with progressive neurologic disease. In 1 horse, Borrelia burgdorferi sensu stricto was identified by polymerase chain reaction amplification of B burgdorferi sensu stricto-specific gene targets (ospA, ospC, flaB, dbpA, arp). Highest spirochetal burdens were in tissues with inflammation, including spinal cord, muscle, and joint capsule. Sequence analysis of ospA, ospC, and flaB revealed 99.9% sequence identity to the respective genes in B burgdorferi strain 297, an isolate from a human case of neuroborreliosis. In both horses, spirochetes were visualized in affected tissues with Steiner silver impregnation and by immunohistochemistry, predominantly within the dense collagenous tissue of the dura mater and leptomeninges.

Interface membrane is the best sample for histological study to diagnose prosthetic joint infection.

The objective of our study was to study which is the most accurate specimen for histological diagnosis of prosthetic joint infections (pseudocapsule or interface membrane). This is a prospective study including hip revision arthroplasties performed from January 2007 to June 2009. Specimens from pseudocapsule and from interface membrane were obtained from each patient. The histology was considered positive for infection when ≥5 neutrophils per high-power field ( × 40) were found. Definitive diagnosis of infection was considered when ≥2 cultures were positive for the same microorganism. According to the definition of infection, patients were classified in two groups: (A) patients with aseptic loosening in whom cultures obtained during surgery were negative and (B) patients with prosthetic joint infection. A total of 69 revisions were included in the study; 57 were classified in group A and 12 in group B. In group B, the percentage of positive interface membrane histology was significantly higher than the percentage of positive pseudocapsule histology (83 vs 42%, P=0.04, Fisher's exact test). The results suggest that periprosthetic interface membrane is the best specimen for the histological diagnosis of prosthetic joint infection.

Histopathological studies of experimental lyme disease in the dog.

Experimental borrelia infection was induced in 62 specific--pathogen-free beagle dogs by exposure to Ixodes scapularis ticks harbouring the spirochaete Borrelia burgdorferi. Clinical signs of Lyme disease occurred in 39/62 dogs, the remaining 23 being subclinically infected. Clinical signs consisted of one to six episodes of transitory lameness with joint swelling and pain, most commonly affecting the elbow or shoulder joints. The polymerase chain reaction and culture demonstrated that the dogs remained infected for up to 581 days. At necropsy, gross findings consisted of lymphadenopathy in the area of tick attachment. Microscopical changes consisted of effusive fibrinosuppurative inflammation or nonsuppurative inflammation, or both, affecting synovial membranes, joint capsules and associated tendon sheaths. Plasma cells dominated areas of chronic inflammation, with CD3(+) T cells being present in lesser numbers. Microscopical signs of arthritis were polyarticular and more widespread than indicated by clinical signs, and most of the subclinically affected animals also had synovitis. In areas of tick attachment to the skin, hyperkeratosis and a mixture of suppurative and nonsuppurative dermatitis were encountered. Lymphadenopathy in superficial lymph nodes resulted from follicular and parafollicular hyperplasia. In 14/62 dogs, lymphoplasmacytic periarteritis and perineuritis were noted, resembling lesions found in human Lyme disease and syphilis, in which an underlying microangiopathy has been proposed.

MRI findings of septic arthritis and associated osteomyelitis in adults.

OBJECTIVE: The purpose of this study was to describe the soft-tissue, synovial, and osseous MRI findings of septic arthritis. MATERIALS AND METHODS: At 1.5 T (T1-weighted, T2-weighted or STIR, and contrast-enhanced images), 50 consecutive cases of septic arthritis were evaluated by two observers for synovial enhancement, perisynovial edema, joint effusion, fluid outpouching, fluid enhancement, and synovial thickening. The marrow was assessed for abnormal signal on T1- and T2-weighted images or after contrast enhancement. We noted whether the marrow signal was diffuse or abnormal in bare areas. MRI findings were compared with microbiologic, clinical, and surgical data and diagnoses. RESULTS: The frequency of MRI findings in septic joints was as follows: synovial enhancement (98%), perisynovial edema (84%), joint effusions (70%), fluid outpouching (53%), fluid enhancement (30%), and synovial thickening (22%). The marrow showed bare area changes (86%), abnormal T2 signal (84%), abnormal gadolinium enhancement (81%), and abnormal T1 signal (66%). Associated osteomyelitis more often showed T1 signal abnormalities and was diffuse. CONCLUSION: Synovial enhancement, perisynovial edema, and joint effusion had the highest correlation with the clinical diagnosis of a septic joint. However, almost a third of patients with septic arthritis lacked an effusion. Abnormal marrow signal-particularly if it was diffuse and seen on T1-weighted images-had the highest association with concomitant osteomyelitis.

On the role of Staphylococcus aureus sortase and sortase-catalyzed surface protein anchoring in murine septic arthritis.

Anchoring of Staphylococcus aureus surface protein to the cell wall is catalyzed by sortase, a transpeptidase. The contribution of staphylococcal surface proteins to establishment of infection was examined using a murine septic arthritis model. Intravenous inoculation of mice with the sortase-deficient mutant S. aureus strain SMK3 did not result in weight loss or severe septic arthritis, in contrast to the parent strain, S. aureus Newman. Direct inoculation of the sortase mutant into joint cavities also failed to cause severe synovitis or erosive arthritis. Furthermore, intravenous inoculation with staphylococci resulted in the rapid clearing of the sortase mutant from the bloodstream. This phenomenon demonstrates the involvement of host neutrophils; when these cells were depleted, sortase mutant staphylococci caused severe systemic infection, although not septic arthritis. These results suggest that sortase mutant staphylococci are significantly less virulent than the parent strain, Newman: the sortase mutant has decreased ability to reach target organs and, once there, to induce an inflammatory response.

Experimental induction of chronic borreliosis in adult dogs exposed to Borrelia burgdorferi-infected ticks and treated with dexamethasone.

OBJECTIVE: To develop a method to experimentally induce Borrelia burgdorferi infection in young adult dogs. ANIMALS: 22 healthy Beagles. PROCEDURE: All dogs were verified to be free of borreliosis. Twenty 6-month-old dogs were exposed to Borrelia burgdorferi-infected adult ticks and treated with dexamethasone for 5 consecutive days. Two dogs not exposed to ticks were treated with dexamethasone and served as negative-control dogs. Clinical signs, results of microbial culture and polymerase chain reaction (PCR) testing, immunologic responses, and gross and histologic lesions were evaluated 9 months after tick exposure. RESULTS: Predominant clinical signs were episodic pyrexia and lameness in 12 of 20 dogs. Infection with B burgdorferi was detected in microbial cultures of skin biopsy specimens and various tissues obtained during necropsy in 19 of 20 dogs and in all 20 dogs by use of a PCR assay. All 20 exposed dogs seroconverted and developed chronic nonsuppurative arthritis. Three dogs also developed mild focal meningitis, 1 dog developed mild focal encephalitis, and 18 dogs developed perineuritis or rare neuritis. Control dogs were seronegative, had negative results for microbial culture and PCR testing, and did not develop lesions. CONCLUSIONS AND CLINICAL RELEVANCE: Use of this technique successfully induced borreliosis in young dogs. Dogs with experimentally induced borreliosis may be useful in evaluating vaccines, chemotherapeutic agents, and the pathogenesis of borreliosis-induced arthritis.

Contamination of banked femoral head allograft: incidence, bacteriology and donor follow up.

BACKGROUND: Allograft donations are not uncommonly found to be contaminated. The issue of contaminated donations from live donors at the time of surgery, and the significance of this to the patient in terms of subsequent sepsis of the arthroplasty, were examined. METHODS: The donations of femoral heads to the Queensland Bone Bank over a 9-year period were reviewed, and the incidence and bacteriology of contamination were detailed. Clinical outcomes were determined for donors who had positive cultures at the time of retrieval and they were compared with those of culture-negative donors. RESULTS: Between March 1987 and February 1996, 232 femoral heads were donated to the Queensland Bone Bank. Four specimens were sent for culture with each femoral head (surface swab of femoral head, acetabular swab, bone biopsy and capsule). In 51 cases, one or more positive cultures were obtained (22% contamination rate). The majority of organisms cultured were Staphylococcus epidermidis. One hundred and seventy donations came from surgery performed at the Princess Alexandra Hospital, and 40 femoral heads were considered contaminated. Deep infection was recorded in one of the 40 cases with contaminated donations and three out of 130 non-contaminated donations had subsequent septic episodes. CONCLUSION: The contamination rate detailed in the present report is higher than in most series. This may be due to the fact that four bacteriological specimens are taken to assess contamination. Two of these specimens are tissue samples which yielded more positive results than did the two swabs. All other series take no more than two bacteriological specimens, which are usually bone swabs. These are shown to have a poor yield of positive cultures. Therefore there is a significant underestimation of contamination rates by other bone banks. This has implications for the recipients of bone from those banks, particularly when the allograft material is not secondarily sterilized. This is important given increasing allograft usage, and the increasing numbers of revision joint arthroplasty and impaction grafting procedures being performed. Sterilization of all bone by irradiation to 25 kGy is recommended.

Detection of human serum antibody to encapsulated strains of Staphylococcus aureus by enzyme-linked immunosorbent assay inhibition test.

A specific and rapid enzyme-linked immunosorbent assay (ELISA) inhibition test was employed for detection of immunoglobulins to Staphylococcus aureus (S. aureus) capsular polysaccharide in human serum. Cap-sular polysaccharide antigens obtained from Smith diffuse (capsular type 2), Reynolds (capsular type 5), or Becker (capsular type 8) strains of S. aureus were added to microplates coated with these strains. Seventy-four patients with open fractures (31 serum samples from those with staphylococcal infections, 10 serum samples from those with non-staphylococcal infections, and 33 serum samples from the non-infected group) and 28 serum samples from healthy controls were then added. The plates were incubated at 37 degreesC for 2 h and the ELISA was performed. The ELISA inhibition assay showed remarkable inhibition with the capsular type 2, 5, and 8 polysaccharides in the 33 serum samples from the non-infected group and in the 28 serum samples from the healthy controls, but low inhibition was observed with the 31 sera with staphylococcal infections. Positive immunoglobulin (Ig)G and IgM titers showed marked inhibition with this assay, but IgA titer were not seen in any samples. These results indicate that the quantitation of human serum antibody against S. aureus capsular polysaccharide by the ELISA inhibition assay is useful for the demonstration of protective activities against S. aureus.