Surgical Management of Periprosthetic Joint Infections in Hip and Knee Megaprostheses.

Periprosthetic joint infection is a feared complication after the megaprosthetic reconstruction of oncologic and non-oncologic bone defects of including the knee or hip joint. Due to the relative rarity of these procedures, however, optimal management is debatable. Considering the expanding use of megaprostheses in revision arthroplasty and the high revision burden in orthopedic oncology, the risk of PJI is likely to increase over the coming years. In this non-systematic review article, we present and discuss current management options and the associated results focusing on studies from the last 15 years and studies from dedicated centers or study groups. The indication, surgical details and results in controlling infection are presented for debridement, antibiotics, irrigation and retention (DAIR) procedure with an exchange of the modular components, single-stage implant exchange, two-stage exchanges and ablative procedures.

Survey on microbial contamination of opened skin care products used for tattooing.

AIMS: Inadequate hygiene measures as well as the use of contaminated inks or non-sterile needles are considered as important infection sources in the process of tattooing. In tattoo parlors and at conventions, it is common practice to apply cosmetic products from bulk packs as lubricants during tattooing and as ointments for tattoo aftercare. The objective of our study was to assess the microbial load of opened skin care products used during tattooing or for tattoo aftercare. METHODS AND RESULTS: First, we established a homogenization method suitable for the microbiological examination of water-immiscible products. To this end, we compared the efficiency of FastPrepTM and Stomacher® homogenizers on artificially contaminated petroleum jelly. FastPrep homogenates revealed significantly higher detection rates (≥97%) compared to Stomacher ones (31%-64%). Second, we investigated 106 cosmetic bulk pack products collected from tattoo artists. After FastPrep homogenization for 30 seconds, total aerobic mesophilic bacteria and the presence of Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans were determined through culture. We also tested for Mycobacteria spp. by qPCR. In total, 4.7% of the cosmetic products under study turned out to be contaminated. CONCLUSION: The observed microbial contamination of opened skin care bulk packs can hold a risk to introduce bacteria into the fresh skin wound resulting from tattooing and may be a risk factor for post-tattoo infections.

Revision TKA with a distal femoral replacement is at high risk of reinfection after two-stage exchange for periprosthetic knee joint infection.

PURPOSE: Megaprosthetic distal femoral reconstruction (DFR) is a limb-salvage procedure to address bone loss following two-stage revision for periprosthetic knee joint infection (PJI). The purpose of this study was to analyze the survival of DFR compared to hinged total knee arthroplasty (TKA). It was hypothesized that DFR was associated with a poorer survival. METHODS: In this retrospective single-center study, 97 subjects who underwent two-stage revision of chronic knee PJI were included. Among these, 41 were DFR. The diagnosis of PJI was established using the Musculoskeletal Infection Society (MSIS) criteria. Implant survival was calculated using Kaplan-Meier method and compared with the log-rank test as well as multivariate Cox regression at a minimum follow-up period of 24 months. RESULTS: The median follow-up period was 59 (interquartile range (IQR) 45-78) months. Overall, 24% (23/97) of patients required revision surgery for infection. The infection-free survival of rotating hinge revision TKA was 93% (95% Confidence Interval (CI) 86-100%) at five years compared to 50% (95% CI 34-66%) for DFR. In multivariate analysis, the risk factors for reinfection were DFR reconstruction (HR 4.7 (95% CI 1-22), p = 0.048), length of megaprosthesis (HR 1.006 (95% CI 1.001-1.012), p = 0.032) and higher BMI (HR 1.066, 95% CI 1.018-1.116), p = 0.007). 10% (4/41) of patients undergoing DFR underwent amputation to treat recurrent infection. CONCLUSION: Megaprosthetic DFR as part of a two-stage exchange for PJI is a salvage treatment that has a high risk for reinfection compared to non-megaprosthetic TKA. Patients must therefore be counseled accordingly. LEVEL OF EVIDENCE: Retrospective observational study, Level IV.

Unsuspected Positive Cultures in Planned Aseptic Revision Knee or Hip Arthroplasty-Risk Factors and Impact on Survivorship.

BACKGROUND: Unsuspected positive cultures (UPCs) may be found at the time of planned aseptic revision surgery and have previously been associated with decreased implant survival. However, reported rates vary greatly, and potential risk factors are widely unknown. This study investigates the rate of UPCs, implant survival, as well as potential risk factors for UPCs and revision. METHODS: This is a retrospective, single-center study on 434 cases of total hip arthroplasty (THA 277) and knee revision arthroplasty (TKA 157) performed between 2010 and 2017. Microbiology culture results, revision-free survival (RFS) and infection-free (IFS) survival, and the potential impact of UPCs as well as patient- or procedure-related risk factors were investigated at a minimum follow-up of 24 months (median 41 months). RESULTS: A total of 37% of patients had a positive culture (160/434). 27% (119/434) had a solitary positive culture, and 9% (41/434) had ≥2 positive cultures of the same species. Most commonly, coagulase-negative staphylococci were cultured. Overall, 32% (13/41) of patients with ≥2 positive cultures underwent revision for infection compared with 6% (17/274) of patients with negative cultures. RFS and IFS were reduced if ≥2 cultures were positive. Single positive cultures had no impact on RFS or IFS. Male sex, elevated serum C-reactive protein, and obesity were independently associated with finding ≥2 UPCs in revision THA. CONCLUSION: Appearance of ≥2 UPCs in aseptic revision leads to reduced RFS and IFS. Males, obese patients, and patients with elevated serum C-reactive protein planned for THA revision should be considered for extended diagnostics to rule out periprosthetic joint infection.

Total joint replacement of the hip and knee in patients with arthrogryposis multiplex congenita: a report of six joints.

INTRODUCTION: Arthrogryposis multiplex congenita (AMC) is a rare congenital condition that leads to severe joint contractures and deformities. As painful joint dysplasia and degeneration might develop over time, total joint replacement (TJR) can be a potential treatment option for these patients. The aim of this study is to investigate functional results, implant survivorship and potential complications in patients with AMC who undergo hip or knee arthroplasty. MATERIALS AND METHODS: We retrospectively identified six TJR in three patients at a single centre performed between 2006 and 2019. The median patient age at surgery was 23 years and the median follow-up period was 69 (IQR 55-99) months. We analysed surgical technique, implant survivorship and complications as well as functional outcome determined by pain reported on the Numerical Rating Scale (NRS), patient-reported outcome scores [Oxford Hip Score (OHS), Harris Hip score (HHS), Oxford Knee Score (OKS)], range of motion and ambulatory status. Depending on data distribution means with ranges and median with interquartile range were compared with the Wilcoxon signed rank test or Student's t test. The level of significance was defined at < 0.05. RESULTS: In hips, the mean range of motion in flexion/extension (52° vs. 85°, p = 0.014) and in rotation (28° vs. 68°, p = 0.02) as well as mean pain score on the NRS (8.5 vs. 0, p = 0.001), OHS (9 vs. 26, p = 0.031) and HHS (17 vs. 52, p = 0.007) significantly improved. In knees, mean range of motion (55° vs. 93°, p = 0.403), mean pain score on the NRS (0 vs. 7) and the OKS (2 vs. 21) also improved. While the ambulatory status did not change, the patients who were wheelchair dependent reported less problems with transfers to a bed or chair and the patient who ambulated reported an improved walking distance. One total knee arthroplasty (TKA) underwent revision for an acute, late infection 155 months following the initial surgery. CONCLUSIONS: TJR is a safe procedure in patients with AMC that effectively improves function and reduces pain irrespective of preoperative ambulatory status.

[Typing and particle analysis of squeaking hip endoprostheses : First histopathological analysis to examine the squeaking pathogenesis of ceramic-on-ceramic bearings]. / Typisierung und Partikelanalyse von Squeaking-Hüftendoprothesen : Erste histopathologische Analyse zur Untersuchung der Squeaking-Pathogenese von Keramik-Keramik-Gleitpaarungen.

BACKGROUND: Since the use of ceramic-on-ceramic (CoC) hip endoprostheses complications in the form of squeaking noises have occasionally occurred. OBJECTIVES: This is the first histopathological analysis of the synovia-like interface membrane (SLIM) of ceramic squeaking hip endoprostheses with the aim to gain new insights into the squeaking pathogenesis. MATERIALS AND METHODS: Seven CoC hip endoprostheses with squeaking pathogenesis are analyzed by SLIM consensus classification, particle algorithm, CD3 quantification, semiquantitative CD68 macrophages, Oil-Red positive macrophages, hemosiderin evaluation and in two cases by energy dispersive X­ray spectroscopy (EDX). RESULTS: In 1733 hip joint prosthesis pathology cases, a squeaking revision incidence of 0.40% was determined. In addition to SLIM type I (1/7), only SLIM type IV (6/7) was detected. 4/7 CoC cases showed combinations of micro, macro and, for the first time, supramacro (166.5 µm) ceramic wear particles. The EDX analysis confirmed the ceramic and an additional metallic abrasion. Increased focal concentrated low inflammatory markers (CD3/CD68) with hemosiderin (5/7) and lipid depositions (Oil-Red positive macrophages) (6/7) occurred. CONCLUSIONS: A pathogenetic connection between SLIM type I/IV and squeaking can be assumed. SLIM types showed a partly light microscopic ceramic particle-dependent, partly independent predominantly low-grade inflammation. Hemosiderin and Oil-Red positive macrophages are signs of synovial tissue damage and indicate biomechanical misload (impingement) and dysfunction as cause of the squeaking pathogenesis.

Microbially competent 3D skin: a test system that reveals insight into host-microbe interactions and their potential toxicological impact.

The skin`s microbiome is predominantly commensalic, harbouring a metabolic potential far exceeding that of its host. While there is clear evidence that bacteria-dependent metabolism of pollutants modulates the toxicity for the host there is still a lack of models for investigating causality of microbiome-associated pathophysiology or toxicity. We now report on a biologically characterised microbial-skin tissue co-culture that allows studying microbe-host interactions for extended periods of time in situ. The system is based on a commercially available 3D skin model. In a proof-of-concept, this model was colonised with single and mixed cultures of two selected skin commensals. Two different methods were used to quantify the bacteria on the surface of the skin models. While Micrococcus luteus established a stable microbial-skin tissue co-culture, Pseudomonas oleovorans maintained slow continuous growth over the 8-day cultivation period. A detailed skin transcriptome analysis showed bacterial colonisation leading to up to 3318 significant changes. Additionally, FACS, ELISA and Western blot analyses were carried out to analyse secretion of cytokines and growth factors. Changes found in colonised skin varied depending on the bacterial species used and comprised immunomodulatory functions, such as secretion of IL-1α/ß, Il-6, antimicrobial peptides and increased gene transcription of IL-10 and TLR2. The colonisation also influenced the secretion of growth factors such as VFGFA and FGF2. Notably, many of these changes have already previously been associated with the presence of skin commensals. Concomitantly, the model gained first insights on the microbiome's influence on skin xenobiotic metabolism (i.e., CYP1A1, CYP1B1 and CYP2D6) and olfactory receptor expression. The system provides urgently needed experimental access for assessing the toxicological impact of microbiome-associated xenobiotic metabolism in situ.

Subtle changes of gray matter volume in fibromyalgia reflect chronic musculoskeletal pain rather than disease-specific effects.

Fibromyalgia syndrome (FMS) is a chronic pain syndrome. Neuroimaging studies provided evidence of altered gray matter volume (GMV) in FMS but, similarly, in chronic pain of other origin as well. Therefore, the purpose of this study was to evaluate the disease specificity of GMV alterations in FMS by direct comparison. Structural MRI data of the brain were acquired in 25 females with FMS and two different control groups: 21 healthy subjects and 23 patients with osteoarthritis. Regional GMVs were compared by voxel-based morphometry and additional ROI-analyses. In conclusion, we did not identify significant GMV alterations in either FMS or OA patients compared to healthy controls when adopting a conservative statistical approach with multiple comparison correction. However, even under a more liberal approach no FMS-specific GMV changes were found because both pain groups presented increased gray matter volumes in the precentral gyrus and decreased GMV in the angular gyrus/middle occipital gyrus and middle temporal gyrus in comparison with healthy controls. Since no differences between both pain groups could be detected cortical GMV changes in FMS should not be interpreted as FMS-specific but might rather reflect changes in chronic pain in general. This previously held notion is confirmed in this study by direct comparison with a control group consisting of another pain disorder.

A single centre study of 41 cases on the use of porous tantalum metal implants in acetabular revision surgery.

BACKGROUND: This study aims at investigating cup survival of porous tantalum revision cups and identifies risk factors for failure. METHODS: We retrospectively reviewed 41 patients treated between 2010 and 2012. Main indications were aseptic loosening in 83% and two-stage exchange after periprosthetic joint infection in 17% of cases. Mean follow-up period was 72 months. Femoral megaprostheses were used in 13% of cases. Most defects were classified as Paprosky 3b (29%). Function was assessed using the Harris Hip score. RESULTS: Aseptic cup survivorship was 80% at 104 months (95% Confidence Interval 67.4-92.4). Overall implant survival was 73%. Major bone loss defects (Paprosky types 2c to 3b) were associated with a significantly higher rate of failure than minor defects (P = 0.002). There were eight cases of aseptic loosening (19.5%) and two of infection (4.9%). Previous surgeries, indication for acetabular revision, patient-related risk factors and use of megaprostheses did not significantly influence implant survival. The Harris Hip Score improved from a median of 40 (Interquartile range 31-45) to 82 (interquartile range 65-88) postoperative (P < 0.0001). CONCLUSIONS: In summary, the use of porous tantalum metal implants in acetabular revision surgery achieves good to excellent short- term and mid-term functional results and an acceptable complication rate relative to the extent of defect and previous surgery. However, one should be aware of potential limitations of the implants in addressing large defects and discontinuity.

Two stage revision with a proximal femur replacement.

BACKGROUND: Despite very good prosthesis retention times, the growing numbers of primary implantations of hip endoprostheses are leading to increasing numbers of revision operations. Periprosthetic infection, particularly in revision implants, often leads to a massive loss of bone stock, so that in a two-stage exchange the only option left is implantation of a megaendoprosthesis. This retrospective study investigated the clinical and functional outcome for patients who received megaendoprostheses in the proximal femur in two-stage exchange procedures. METHODS: Forty-nine patients were treated between 1996 and 2014 (mean age 71 years, mean follow-up period 52 months). Microorganisms were isolated intraoperatively in 44 patients (89.9%). The reinfection rate was documented in patients who did not undergo any further revision surgery due to mechanical failure (primary) and in patients who had subsequent revisions after reimplantation and subsequent reinfection (secondary). RESULTS: The mean C-reactive protein level at the time of reimplantation was 1.25 mg/dL (range 0.5-3.4). The primary success rate with curative treatment for prosthetic joint infection was 92% (four of 49 patients). The secondary success rate with infection revision cases was 82% (three of 17 revision cases). The mean Harris hip score was 69 (range 36-94). The majority of patients needed different types of walking aid or even wheelchairs, and only 50% of the patients were able to walk outside. CONCLUSIONS: Reinfections occurred in only 8% of patients who underwent two-stage exchanges with a proximal femur replacement. When revision surgery for the proximal femur replacement was required for mechanical reasons, however, the associated reinfections increased the reinfection rate to 18%. Proximal femur replacement achieves a clear reduction in pain, maintenance of leg length, and restoration of limited mobility, and the procedure thus represents a clear alternative to the extensive Girdlestone procedure, which is even more immobilising, or mutilating amputation.

Acetabular Erosion After Bipolar Hemiarthroplasty in Proximal Femoral Replacement for Malignant Bone Tumors.

BACKGROUND: Hemiarthroplasty megaprosthetic proximal femur reconstruction after tumor resection is a widespread procedure in orthopedic oncology. One potential complication is acetabular wear requiring secondary acetabular revision. The study's purpose is to investigate prevalence of acetabular erosion, secondary revisions, and potential risk factors. METHODS: We retrospectively identified 112 patients who underwent proximal femur replacement after resection of a malignant bone tumor and had radiological follow-up longer than 12 months. Patient demographic, surgical, and oncologic factors were recorded, acetabular wear was measured using the classification proposed by Baker, and prosthetic failure was classified using the International Society on Limb Salvage classification. Functional assessment was performed using the Musculoskeletal Tumor Society Score and Harris Hip Score. RESULTS: Prevalence of acetabular wear was 28.6%. Secondary conversion to total hip arthroplasty was required in 5 patients (4.6%), all treated for primary bone tumors. No patient treated for metastatic tumor had higher grade acetabular wear or required revision. Significant risk factors for the development of acetabular wear were age under 40 (P = .035) and longer follow-up (63 vs 43 months, P = .004). Other patient, surgical, or adjuvant treatment-related factors were not associated with acetabular revision or acetabular wear. The dislocation rate in the patient cohort was 0.9%. CONCLUSION: Bipolar hemiarthroplasty proximal femoral replacement represents a durable reconstruction after tumor resection. Hip instability is rare. Acetabular erosion is rare and can be successfully treated with conversion to total hip arthroplasty. Young patients with long-term survival over 10 years are at risk. In reconstruction for metastases, instability and acetabular wear are rare.

Molecular Tracing to Find Source of Protracted Invasive Listeriosis Outbreak, Southern Germany, 2012-2016.

We investigated 543 Listeria monocytogenes isolates from food having a temporal and spatial distribution compatible with that of the invasive listeriosis outbreak occurring 2012-2016 in southern Germany. Using forensic microbiology, we identified several products from 1 manufacturer contaminated with the outbreak genotype. Continuous molecular surveillance of food isolates could prevent such outbreaks.

Differential detection of pathogenic Yersinia spp. by fluorescence in situ hybridization.

Yersinia enterocolitica, Y. pseudotuberculosis and Y. pestis are pathogens of major medical importance, which are responsible for a considerable number of infections every year. The detection of these species still relies on cultural methods, which are slow, labour intensive and often hampered by the presence of high amounts of accompanying flora. In this study, fluorescence in situ hybridization (FISH) was used to develop a fast, sensitive and reliable alternative to detect viable bacteria in food. For this purpose, highly specific probes targeting the 16S and 23S ribosomal RNA were employed to differentially detect each of the three species. In order to enable the differentiation of single nucleotide polymorphisms (SNPs), suitable competitor oligonucleotides and locked nucleic acids (LNAs) were used. Starved cells still showed a strong signal and a direct viable count (DVC) approach combined with FISH optimized live/dead discrimination. Sensitivity of the FISH test was high and even a single cell per gram of spiked minced pork meat could be detected within a day, demonstrating the applicability to identify foodborne hazards at an early stage. In conclusion, the established FISH tests proved to be promising tools to compensate existing drawbacks of the conventional cultural detection of these important zoonotic agents.

Silver-coated megaprostheses: review of the literature.

Periprosthetic infection remains one of the most serious complications following megaendoprostheses. Despite a large number of preventive measures that have been introduced in recent years, it has not been possible to further reduce the rate of periprosthetic infection. With regard to metallic modification of implants, silver in particular has been regarded as highly promising, since silver particles combine a high degree of antimicrobial activity with a low level of human toxicity. This review provides an overview of the history of the use of silver as an antimicrobial agent, its mechanism of action, and its clinical application in the field of megaendoprosthetics. The benefits of silver-coated prostheses could not be confirmed until now. However, a large number of retrospective studies suggest that the rate of periprosthetic infections could be reduced by using silver-coated megaprostheses.

Rapid characterisation of Klebsiella oxytoca isolates from contaminated liquid hand soap using mass spectrometry, FTIR and Raman spectroscopy.

Microbiological monitoring of consumer products and the efficiency of early warning systems and outbreak investigations depend on the rapid identification and strain characterisation of pathogens posing risks to the health and safety of consumers. This study evaluates the potential of three rapid analytical techniques for identification and subtyping of bacterial isolates obtained from a liquid hand soap product, which has been recalled and reported through the EU RAPEX system due to its severe bacterial contamination. Ten isolates recovered from two bottles of the product were identified as Klebsiella oxytoca and subtyped using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI TOF MS), near-infrared Fourier transform (NIR FT) Raman spectroscopy and Fourier transform infrared (FTIR) spectroscopy. Comparison of the classification results obtained by these phenotype-based techniques with outcomes of the DNA-based methods pulsed-field gel electrophoresis (PFGE), multi-locus sequence typing (MLST) and single nucleotide polymorphism (SNP) analysis of whole-genome sequencing (WGS) data revealed a high level of concordance. In conclusion, a set of analytical techniques might be useful for rapid, reliable and cost-effective microbial typing to ensure safe consumer products and allow source tracking.

Forequarter amputation: a safe rescue procedure in a curative and palliative setting in high-grade malignoma of the shoulder girdle.

BACKGROUND: Forequarter amputation (FQA) is a surgical treatment of tumors in the upper extremity and shoulder girdle that infiltrate the neurovascular bundles. In both curative and palliative settings, FQA can serve as an effective oncological treatment. METHODS: This study presents the FQA-related data of 30 patients (mean age 50 years) treated between 2000 and 2012. Their medical condition was high-grade bone or soft tissue sarcoma in 26 and high-grade carcinoma in four cases. RESULTS: Mean operation time was 119 min. One major and five minor complications occurred in the postoperative period. Resection margins were wide in 91 % of patients. Local recurrence was observed in four patients after 19 months on average. Patients treated with curative intention had a 5-year survival chance of 39 %. Average survival after palliative amputation was 11 months. CONCLUSIONS: FQA provides an opportunity for adequate oncological margins in large tumors, while offering relief from tumor-induced distress in palliative situations.

Knee salvage in revision arthroplasty after massive bone loss of the femur condyles (≥Engh III) with a single-modular-hinged knee revision implant.

PURPOSE: Massive bone loss of the femur condyles in revision arthroplasty often requires modular-hinged revision implants to restore a stable joint situation. In the present series, the outcome after knee revision surgery using a single modular-hinged revision implant in patients with severe bone defects (>Engh III) is investigated. METHODS: Sixty patients with severe bone defects (≥Engh III) after failed primary and revision knee arthroplasty were included. Medium follow-up was 47 (range 10-84) months after knee revision surgery. Medium patient age was 70 (range 33-87) years at the time of surgery. An average of 2.3 prior knee operations per patient was performed. 70 % of the patients required the knee revision implant after two-stage revision because of a deep implant infection. RESULTS: Estimated 5 year extremity survival was 95 and 65 % implant survival. Reasons for implant revision in decreasing order were reinfection (30 %), aseptic loosening (13 %), and periprosthetic fracture (9.8 %). The average active range of motion in the knee joint was 88° (range 40°-115°) for flexion. An extension deficit of a mean of -6° was (range -50-5° hyper-extension) observed. Patient age influenced the functional results significantly in terms of reduced walking distances and decreased modified WOMAC score. CONCLUSION: In consideration of this complex study, population acceptable functional results can be achieved using a modular knee revision endoprosthesis. In younger patients (<60 years), satisfying results in terms of walking ability and overall satisfactory can be expected. The outcome in older multimorbid patients is worse. Yet, operation in these patients can be feasible to restore enough mobility for daily household activities.

Two-stage revision arthroplasty for periprosthetic joint infections: What is the value of cultures and white cell count in synovial fluid and CRP in serum before second stage reimplantation?

INTRODUCTION: Besides CRP in serum, white cell counts and cultures of synovial fluid are routinely used to detect periprosthetic joint infections. But the sensitivities of these parameters do vary from 12 to 100 %. In two stage revision arthroplasty before the second stage surgeons have to decide if reimplantation is justified. Therefore, we investigated the value of cultures and white cell count from the synovial fluid with a polymethyl methacrylate spacer in place and CRP in serum before reimplantation to detect persistent infection in a standardized setting. METHODS: 115 patients with a two-stage revision hip or knee arthroplasty were included in this study. All patients had an antibiotic loaded polymethylmethacrylate spacer. Retrospectively synovial cultures, white blood count in synovial fluid and CRP in serum were assessed before reimplantation. RESULTS: The sensitivity of the synovial cultures was 5 % (95 % CI 0.13-24.87), with a specificity of 99 % (95 % CI 94.27-99.97). For white blood count in synovial fluid the sensitivity was 31.3 %, specificity was 39.1 %. Sensitivity for CRP in serum was 42.10 %, specificity was 84.21 %. CONCLUSION: Cultures from synovial fluid and white blood count in synovial fluid and CRP seem to be uncertain parameters to exclude persistent infection. We do not recommend joint aspiration before reimplantation anymore. Further research is necessary to find other markers to confirm or exclude persistent infection.

FISHing for bacteria in food--a promising tool for the reliable detection of pathogenic bacteria?

Foodborne pathogens cause millions of infections every year and are responsible for considerable economic losses worldwide. The current gold standard for the detection of bacterial pathogens in food is still the conventional cultivation following standardized and generally accepted protocols. However, these methods are time-consuming and do not provide fast information about food contaminations and thus are limited in their ability to protect consumers in time from potential microbial hazards. Fluorescence in situ hybridization (FISH) represents a rapid and highly specific technique for whole-cell detection. This review aims to summarize the current data on FISH-testing for the detection of pathogenic bacteria in different food matrices and to evaluate its suitability for the implementation in routine testing. In this context, the use of FISH in different matrices and their pretreatment will be presented, the sensitivity and specificity of FISH tests will be considered and the need for automation shall be discussed as well as the use of technological improvements to overcome current hurdles for a broad application in monitoring food safety. In addition, the overall economical feasibility will be assessed in a rough calculation of costs, and strengths and weaknesses of FISH are considered in comparison with traditional and well-established detection methods.

Peptaibol, secondary-metabolite, and hydrophobin pattern of commercial biocontrol agents formulated with species of the Trichoderma harzianum complex.

The production of bioactive polypeptides (peptaibiotics) in vivo is a sophisticated adaptation strategy of both mycoparasitic and saprotrophic Trichoderma species for colonizing and defending their natural habitats. This feature is of major practical importance, as the detection of peptaibiotics in plant-protective Trichoderma species, which are successfully used against economically relevant bacterial and fungal plant pathogens, certainly contributes to a better understanding of these complex antagonistic interactions. We analyzed five commercial biocontrol agents (BCAs), namely Canna(®) , Trichosan(®) , Vitalin(®) , Promot(®) WP, and TrichoMax(®) , formulated with recently described species of the Trichoderma harzianum complex, viz. T. afroharzianum, T. simmonsii, and T. guizhouense. By using the well-established, HPLC/MS-based peptaibiomics approach, it could unequivocally be demonstrated that all of these formulations contained new and recurrent peptaibols, i.e., peptaibiotics carrying an acetylated N-terminus, the C-terminus of which is reduced to a 1,2-amino alcohol. Their chain lengths, including the amino alcohol, were 11, 14, and 18 residues, respectively. Peptaibols were also to be the dominating secondary metabolites in plate cultures of the four strains obtained from four of the Trichoderma- based BCAs, contributing 95% of the UHPLC-UV/VIS peak areas and 99% of the total ion count MS peak area from solid media. Furthermore, species-specific hydrophobins, as well as non-peptaibiotic secondary metabolites, were detected, the latter being known for their antifungal, siderophore, or plant-growth-promoting activities. Notably, none of the isolates produced low-molecular weight mycotoxins.