Efficacy of sodium hypochlorite against multidrug-resistant Gram-negative bacteria.

BACKGROUND: Increased antimicrobial resistance has been observed among many bacteria leading to treatment failures in human and veterinary medicine. Disinfection is a prerequisite for infection control and prevention in healthcare settings. Chlorine compounds are cost-effective and accessible worldwide. AIM: To determine the efficacy of sodium hypochlorite (NaOCl) against multidrug-resistant Gram-negative bacteria (MDR-GNB). METHODS: Minimum inhibitory concentrations (MICs) were determined using broth macro-dilution. Bactericidal efficacy was measured by qualitative and quantitative suspension tests followed by practical tests without mechanical action on stainless steel carriers. The guidelines of the German Association for Applied Hygiene were followed. FINDINGS: Results varied remarkably depending on the method. MICs were 0.1% or 0.2% NaOCl. Qualitative suspension tests revealed up to 500-fold lower bactericidal concentrations. Pseudomonas aeruginosa (P = 0.0025) was significantly less susceptible in these tests whereas quantitative suspension tests revealed no significant differences between strains (P > 0.05). Practical tests determined bactericidal concentrations of 0.8-0.32% NaOCl at 1 min of contact and even lower concentrations for longer contact times. At 1 min, five Klebsiella were significantly less susceptible (P = 0.0124), whereas the lower susceptibility of P. aeruginosa was not confirmed. Organic load inhibited bactericidal activity significantly, whereas contact time had a marginal effect. Differing test results underline that MIC determination and qualitative suspension tests may be insufficient approaches to evaluate bacterial susceptibility or resistance. CONCLUSION: NaOCl efficiently reduced Pseudomonas aeruginosa, Acinetobacter spp., and Klebsiella spp., most notably in the absence of organic matter. Strain- and species-specific differences in susceptibility were noticed, but in general MDR-GNB revealed no higher tolerance to NaOCl.

[Infections of hip and knee endoprostheses. Spectrum of pathogens and the role of multiresistant bacteria]. / Infektionen von Hüft- und Knieendoprothesen. Erregerspektrum und die Rolle multiresistenter Bakterien.

BACKGROUND: Because of the rise in primary implantations in elective knee and hip arthroplasty, the number of complications, particularly due to prosthetic infections has increased. Partly due to multimorbidities, an increase in geriatric patients and often unnecessary use of antibiotics, a change in the spectrum of bacteria with an increase in multi-drug resistant pathogens is to be expected. For physicians this creates not only new medical and economic but also sociopolitical challenges. QUESTION: Has the spectrum of bacteria in prosthetic joint infections after total hip arthroplasty (THA) and total knee arthroplasty (TKA) changed during the 12-year period 2001-2012 in our hospital and what role do multi-drug resistant bacteria play? INVESTIGATION COLLECTIVE: A total of 320 patients with prosthetic joint infections (PJI) following TKA or THA could be identified and were included in this study. The sample consisted of 172 patients with an infection after THA (56 % females n = 96 and 44 % males n = 76) with a mean age of 70.9 years (range 39-92 years) and 148 patients with an infection after TKA (55 % females n = 82 and 45 % males n = 66) with a mean age of 70.7 years (range 15-87 years). The bacteria detected and the development over the course of time were evaluated. RESULTS: An increase was found in the occurrence of coagulase negative staphylococci (CNS), in particular Staphylococcus epidermidis (2001-2003 n = 10 and 2010-2012 n = 27). The proportion of oxacillin and methicillin-resistant Staphylococcus epidermidis (MRSE) was also found to increase (0 % in 2001-2003 and 74 % in 2010-2012). A substantial increase in methicillin-resistant Staphylococcus aureus (MRSA) infections could not be found and there was a tendency towards reduction in the total number of Staphylococcus aureus infections. A total of five extended spectrum beta-lactamase (ESBL)-producing bacteria were isolated. CONCLUSION: The spectrum of bacteria has only slightly changed over the years from 2001 to 2012, whereby an increase was only found in the number of CNS infections. Multi-drug resistant bacteria, in particular MRSE have increased. The changes in MRSE found in this study do not appear to warrant a general rethinking of antibiotic prophylaxis.

Large hospital outbreak of KPC-2-producing Klebsiella pneumoniae: investigating mortality and the impact of screening for KPC-2 with polymerase chain reaction.

BACKGROUND: Multi-drug-resistant Klebsiella pneumoniae carbapenemase (KPC)-2-producing K. pneumoniae are an increasing cause of healthcare-associated infections worldwide. AIMS: To investigate the impact of clinical infection on mortality, and examine the effect of use of KPC-2-specific polymerase chain reaction (PCR) on the time to contact isolation during an outbreak. METHODS: Cases were defined as patients clinically infected or colonized with KPC-2-producing K. pneumoniae between June 2010 and July 2012. Cases were described by demographic and health characteristics, and the association between infection and mortality, adjusted for comorbidities and demographic characteristics, was determined using Poisson regression with robust standard errors. A comparison was made between the time to contact isolation with a culture-based method and PCR using Wilcoxon's rank sum test. FINDINGS: Of 72 cases detected, 17 (24%) had undergone transplantation and 21 (29%) had a malignancy. Overall, 35 (49%) cases were clinically infected, with pneumonia and sepsis being the most common infections. Infection was an independent risk factor for mortality (risk ratio 1.67, 95% confidence interval 0.99-2.82). The median time to contact isolation was 1.5 days (range 0-21 days) using PCR and 5.0 days (range 0-39 days) using culture-based methods (P = 0.003). Intermittent negative tests were observed in 48% (14/29) of cases tested using culture-based methods. CONCLUSION: KPC-2-producing K. pneumoniae mainly affect severely ill patients. Half of the cases developed clinical infection, associated with increased risk of death. As PCR accelerates isolation and provides the opportunity for preventive measures in colonized cases, its use should be implemented promptly during outbreaks. Further studies are needed to enhance knowledge about KPC detection patterns and to adjust screening guidelines.

Review of the prevalence and drug resistance of tuberculosis in prisons: a hidden epidemic.

SUMMARY The prison setting has been often cited as a possible reservoir of tuberculosis (TB) including multidrug-resistant (MDR)-TB. This is particularly true in low-income, high TB prevalence countries in Sub-Saharan Africa. A systemic literature review was done to assess the prevalence, drug resistance and risk factors for acquiring TB in the prison population. Our review indicated a high prevalence of TB in prisons which is reported to be 3- to 1000-fold higher than that found in the civilian population, indicating evidence and the need for public health policy formulation. In addition, high levels of MDR and extensively drug-resistant (XDR)-TB have been reported from prisons, which is a warning call to review prison TB control strategy. Multiple risk factors such as overcrowding, poor ventilation, malnutrition, human immunodeficiency virus (HIV), and others have fuelled the spread of TB in prisons. Furthermore, the impact extends beyond the prison walls; it affects the civilian population, because family visits, prison staff, and members of the judiciary system could be potential portals of exit for TB transmission. The health of prisoners is a neglected political and scientific issue. Within these background conditions, it is suggested that political leaders and scientific communities should work together and give special attention to the control of TB and MDR-TB in prisons. If not, TB in prisons will remain a neglected global problem and threatens national and international TB control programmes. Further researches are required on the prevalence and drug resistance of smear-negative TB in prisons. In addition, evidence of the circulating strains and transmission dynamics inside prisons is also warranted.

[Pitfalls in determining resistance]. / Fallstricke bei der Resistenzbestimmung.

Selection of the optimal antimicrobial therapy is often crucial for the morbidity and mortality associated with an infection. A reliable antibiogram provides the basis for antibiotic therapy optimization. In case of life-threatening infections, minimal inhibitory concentration values should be available so that, in conjunction with knowledge of the pharmacokinetic properties of the respective antibiotics, a rational selection addressing the individual case is feasible. If only categorized results (susceptible, intermediate, resistant: S-I-R) are available, they should be established according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines in order to avoid therapeutic failures.

Colonization of liver transplant recipients with KPC-producing Klebsiella pneumoniae is associated with high infection rates and excess mortality: a case-control analysis.

PURPOSE: From mid-2010 to early 2013 there was a large single-center (Leipzig University Hospital, Germany) outbreak of Klebsiella pneumoniae carbapenemase (KPC) type 2 producing K. pneumoniae (KPC-2-KP) involving a total of 103 patients. The aim of this study was to compare KPC-positive liver transplant recipients (LTR) and KPC-negative controls to determine both the relative risk of infection following colonization with KPC-2-KP and the case fatality rate associated with KPC-2-KP. METHODS: The study cohort of this retrospective observational study comprised nine patients who had undergone orthotopic liver transplantation (LTx) (median age of 52 years, range 28-73 years) with confirmed evidence of colonization with KPC-2-KP. The data from these nine LTR were matched to 18 LTR (1:2) in whom carbapenem-resistant pathogens were not present and compared for clinical outcomes. RESULTS: Of these nine cases, eight (89 %) progressed to infection due to KPC-2-KP, and five (56 %) were confirmed to have bloodstream infection with KPC-2-KP. Matched-pair analysis of KPC-positive LTR and KPC-negative controls revealed a substantially increased relative risk of 7.0 (95 % confidence interval 1.8-27.1) for fatal infection with KPC-2-producing K. pneumoniae after transplantation with a mortality rate of 78 % (vs. 11 %, p = 0.001). CONCLUSIONS: Colonization with KPC-2-KP in LTR leads to high infection rates and excess mortality. Therefore, frequent screening for carbapenem-resistant bacteria in patients on LTx waiting lists appears to be mandatory in an outbreak setting. Patients with evidence of persistent colonization with KPC-producing pathogens should be evaluated with extreme caution for LTx.

EUCAST expert rules in antimicrobial susceptibility testing.

EUCAST expert rules have been developed to assist clinical microbiologists and describe actions to be taken in response to specific antimicrobial susceptibility test results. They include recommendations on reporting, such as inferring susceptibility to other agents from results with one, suppression of results that may be inappropriate, and editing of results from susceptible to intermediate or resistant or from intermediate to resistant on the basis of an inferred resistance mechanism. They are based on current clinical and/or microbiological evidence. EUCAST expert rules also include intrinsic resistance phenotypes and exceptional resistance phenotypes, which have not yet been reported or are very rare. The applicability of EUCAST expert rules depends on the MIC breakpoints used to define the rules. Setting appropriate clinical breakpoints, based on treating patients and not on the detection of resistance mechanisms, may lead to modification of some expert rules in the future.

First- and second-line anti-tuberculosis drug resistance in Northwest Ethiopia.

SETTING: Gondar Hospital, Gondar Health Centre, Metemma Hospital, Bahir Dar Hospital and Debre Markos Hospital in Northwest Ethiopia. OBJECTIVE: To assess the level of and risk factors for first- and second-line drug resistance among tuberculosis (TB) patients. DESIGN: Drug susceptibility testing (DST) against first-line drugs, including isoniazid (INH), rifampicin (RMP), streptomycin (SM), ethambutol (EMB) and pyrazinamide (PZA), was performed using the BacT/ALERT 3D system. DST against second-line drugs, including fluoroquinolones and aminoglycocides/cyclic peptides, was performed using GenoType MTBDRsl. RESULTS: Of 260 Mycobacterium tuberculosis isolates, 41 (15.8%) were resistant to at least one first-line drug, 13 (5.0%) were multidrug-resistant (MDR) and 9 (3.5%) were resistant to all first-line drugs. Any resistance to INH, RMP, SM, EMB and PZA was respectively 36 (13.8%), 15 (5.8%), 26 (10.0%), 19 (7.3%) and 12 (4.6%). Of 214 new and 46 previously treated cases, respectively 8 (3.7%) and 5 (10.9%) were MDR. All isolates were susceptible to all second-line drugs. CONCLUSION: A substantial number of new and previously treated cases harbour MDR-TB. We recommend DST at least for previously treated cases, patients who remain smear-positive at the end of the second month of treatment and patients in close contact with MDR-TB cases. Improved infection control measures need to be implemented in Ethiopia.

A real-time PCR assay for the differentiation of Candida species.

AIMS: We established a real-time PCR assay for the detection and strain identification of Candida species and demonstrated the ability to differentiate between Candida albicans the most common species, and also Candida parapsilosis, Candida glabrata, Candida tropicalis and Candida dubliniensis by LightCycler PCR and melting curve analysis. METHODS AND RESULTS: The DNA isolation from cultures and serum was established using the QIAmp Tissue Kit. The sensitivity of the assay was ≥ 2 genome equivalents/assay. It was possible to differentiate all investigated Candida species by melting curve analysis, and no cross-reaction to human DNA or Aspergillus species could be observed. CONCLUSIONS: The established real-time PCR assay is a useful tool for the rapid identification of Candida species and a base technology for more complex PCR assays. SIGNIFICANCE AND IMPACT OF THE STUDY: We carried out initial steps in validation of a PCR assay for the detection and differentiation of medically relevant Candida species. The PCR was improved by generating PCR standards, additional generation of melting curves for species identification and the possibility to investigate different specimens simultaneously.

[Tuberculosis and travelling]. / Tuberkulose und Reisen.

Tuberculosis (TB) in Germany in the year 2007 with 5020 reported cases (incidence: 6.1 newly diagnosed cases per 100 000 inhabitants) was continuously in decline. 43.1 % of these persons were from countries with a higher TB incidence as compared to Germany. However, not only migration but also personal journeys from low- to high-incidence countries carries an increased risk of infection with M. tuberculosis (MTB). An early active TB follows only rarely, more common, however, is a latent TB infection (LTBI). Not only the active form of TB but also LTBI, with a potential for reactivation years or decades later, can be of enormous relevance for the individual and the social environment. The early detection of an MTB infection and its possible sequelae are decisive for a continued successful battle against tuberculous diseases, especially in view of increasing travel activities.

Rapid identification of oral anaerobic bacteria cultivated from subgingival biofilm by MALDI-TOF-MS.

INTRODUCTION: To facilitate the identification of anaerobes cultivated from periodontal disease, whole cell bacterial identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was evaluated. METHODS: A total of 84 strains (nine reference strains and 75 recent clinical isolates from 33 patients with aggressive periodontitis) previously identified with phenotypic methods were used. All the references and 10 clinical isolates belonging to the same species as the reference strains were genotypically identified by sequence analysis of the 16S ribosomal RNA gene. All the strains were then analyzed using MALDI-TOF-MS. RESULTS: The reference strains of anaerobic bacteria used showed characteristic MALDI-TOF-MS spectra with peaks between m/z 2000 and up to about m/z 13,000. On visual inspection, the similarity of spectra produced by strains of a single genus could be recognized. Obvious differences between spectra produced by strains of different species were also easily noticed. The reproducibility of the method was proved by the similarity of spectra belonging to the same species. The spectra of the Prevotella intermedia strains identified with MALDI clustered together and clustered separately from the spectra of Prevotella nigrescens, proving that MALDI-TOF-MS is an accurate method that is capable of separating these two species. The quality of clustering was characterized by calculating an inconsistency coefficient (Mathworks:/Matlab Reference Manual v2007a/, Statistical toolbox). CONCLUSION: Our results suggest that MALDI-TOF-MS might become a useful method for the identification of anaerobic bacteria, especially for those that cannot be readily identified by biochemical analysis. It may become an attractive system even for the routine identification of clinical isolates.

High-yield amplification of Cryptosporidium parvum in interferon gamma receptor knockout mice.

To date, large-scale production of Cryptosporidium parvum oocysts has only been achieved by amplification in neonatal calves and sheep. Many laboratories currently depend on supplies from external sources and store oocysts for prolonged periods which results in progressive loss of viability. Six to 8-week-old interferon gamma receptor knockout (IFN gamma R-KO) mice on a C57BL/6 background were inoculated by gavage (2000 oocysts/animal). Fecal pellets were collected daily from 7 days post-infection (p.i.) up to 2 weeks p.i. Intestinal oocyst yield was assessed at days 11, 12 and 14 p.i. by homogenization of intestinal tissues. Ether extraction and one or more NaCl flotations were used to purify oocysts. Total recoveries averaged 2.6 x 10(6) oocysts/mouse from fecal material and 3.8 x 10(7) oocysts/mouse from intestinal tissues. Overall, 2.3 x 10(9) purified oocysts were obtained from 60 mice. Recovered oocysts were capable of sporulation and were shown to be infectious both in vitro and in vivo. Oocyst amplification was achieved in only 11-14 days with minimal expense. The simplicity of this method presents a practical alternative for the routine passage, maintenance and storage of C. parvum in biomedical laboratories.

[Clinical features and diagnosis of bronchopulmonary infections in the elderly]. / Klinik und Diagnose bronchopulmonaler Infektionen im Alter.

Diseases of the lung are one of the main causes of morbidity and mortality in the elderly. The risk of respiratory infections is increased due to structural changes, malnutrition, co-morbidity, and a variety of other factors. Bacterial and viral pathogens cause acute bronchitis and exacerbations of chronic bronchitis (AECB). Community acquired pneumonias (CAP) show a different spectrum of pathogens and clinical course in comparison to nosocomial pneumonias (hospital acquired pneumonia, HAP). Institutionalised patients are at risk of a health care associated pneumonia (HCAP), with often a different spectrum of pathogens in comparison to CAP and HAP. Elderly patients with cerebrovascular disease and impairment of swallowing or cough reflexes often suffer from aspiration pneumonias. The mortality is highest in the elderly, comorbid, and immunocompromised patient with nosocomial pneumonia. Important preventive measures include influenza and pneumococcal vaccination, avoidance of immobility, oral hygiene, and sufficient nutrition.

Comparative analysis of antimicrobial susceptibility among organisms from France, Germany, Italy, Spain and the UK as part of the tigecycline evaluation and surveillance trial.

As part of the tigecycline evaluation and surveillance trial (TEST), bacterial isolates were collected from 39 centres in France, Germany, Italy, Spain and the UK between January 2004 and August 2006. Antimicrobial susceptibilities were determined according to CLSI guidelines. Italy had the highest rate of methicillin-resistant Staphylococcus aureus (36.4%), and was the only country to report vancomycin-resistant Enterococcus faecalis (8.6%). Tigecycline was the only agent to which all Gram-positive isolates were susceptible. For many of the Gram-negative organisms collected, antimicrobial susceptibilities were lowest among isolates from Italy and highest among isolates from Spain. The notable exception was Acinetobacter baumannii, where the poorest susceptibility profile was among isolates from Spain. For A. baumannii, MIC(90)s of imipenem varied from 1 mg/L for isolates in France and Germany to > or =32 mg/L for isolates from Italy and Spain. Tigecycline was the only agent to maintain an MIC(90) of < or =1 mg/L against isolates from all five countries. The in-vitro activity of tigecycline against both Gram-positive and Gram-negative isolates may make it valuable in the treatment of hospital infections, including those caused by otherwise antimicrobial-resistant organisms.

Rapid identification of viridans streptococci by mass spectrometric discrimination.

Viridans streptococci (VS) are responsible for several systemic diseases, such as endocarditis, abscesses, and septicemia. Unfortunately, species identification by conventional methods seems to be more difficult than species identification of other groups of bacteria. The aim of the present study was to evaluate the use of cell matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) for the rapid identification of 10 different species of VS. A total of 99 VS clinical isolates, 10 reference strains, and 20 strains from our in-house culture collection were analyzed by MALDI-TOF-MS. To evaluate the mass-spectrometric discrimination results, all strains were identified in parallel by phenotypic and genotypic methods. MALDI-TOF-MS identified 71 isolates as the mitis group, 23 as the anginosus group, and 5 as Streptococcus salivarius. Comparison of the species identification results obtained by the MALDI-TOF-MS analyses and with the phenotypic/genotypic identification systems showed 100% consistency at the species level. Thus, MALDI-TOF-MS seems to be a rapid and reliable method for the identification of species of VS from clinical samples.

Biocompatibility of bacterial contaminated prosthetic meshes and porcine dermal collagen used to repair abdominal wall defects.

BACKGROUND AND AIMS: A contaminated or infected surgical site is considered a contraindication for the use of the nonabsorbable alloplastic materials employed to repair abdominal wall defects. Therefore, the biocompatibility of new prosthetic materials was investigated. MATERIALS AND METHODS: Meshes measuring 1.5x1.5 cm made of conventional and titanium-coated polypropylene, polyglycol, or porcine dermal collagen were implanted under the abdominal wall of 96 rats (eight groups of 12 animals each) employing the inlay technique. Implantation of all four materials was performed both under semisterile conditions and bacterial contamination of the mesh. The meshes were explanted after 28 days. RESULTS: All the materials implanted under semisterile conditions were incorporated into the abdominal wall with only few intraabdominal adhesions (mean adhesion scores: 1.0, 1.2, 1.0, 0.8 points, respectively, not significant). With the porcine dermal collagen, proliferation rate and the proportion of inflammatory cells were statistically lower (p<0.01). In the bacterial contamination group, all meshes were associated with a suppurating infection and strong adhesions between the bowel and mesh, which were most prominent in the case of dermal collagen (mean adhesion scores: 1.6, 1.7, 1.7, and 1.9 points, respectively, not significant). In this group, two animals died of peritonitis. In comparison with the other materials, the proliferation rate was significantly elevated (p=0.03). No significant differences were seen between the other materials employed. CONCLUSION: Irrespective of the material employed, implantation of alloplastic meshes in an abdominal wall contaminated with bacteria, is associated with suppurating infections, in particular in the case of the membrane-like porcine dermal collagen. Nonabsorbable alloplastic meshes and dermal skin grafts should therefore not be used to repair infected abdominal wall defects.

Two decades of imipenem therapy.

Imipenem, the first carbapenem discovered, was developed more than two decades ago in response to an unmet need for a highly potent, broad-spectrum antimicrobial agent with a strong safety profile. It has since been used to treat more than 26 million patients. In an era where antibiotic use has driven antibiotic resistance, choosing appropriate initial therapy for serious infection is critical. Appropriate antibiotic regimens must cover all likely pathogens, be administered promptly at the correct dosage and dosing interval, be well tolerated and prevent the emergence of resistance. While imipenem was initially reserved for use in intractable, serious infections, the benefits of early aggressive therapy are now known, making imipenem a core agent in de-escalation therapy due to proven efficacy and safety for indications such as nosocomial pneumonia, intra-abdominal infection, sepsis and febrile neutropenia. De-escalation therapy with an agent such as imipenem minimizes resistance development by initiating aggressive initial treatment and then tailoring therapy based on patient response and culture results, switching to a less expensive, narrower spectrum antibiotic regimen or shortening the duration of therapy. Imipenem has maintained sustained clinical efficacy, tolerability and in vitro activity against important bacterial pathogens for two decades. We review the factors that continue to make imipenem as appropriate an agent for de-escalation therapy now as it was 20 years ago.

Characterization of the Bacteroides fragilis pathogenicity island in human blood culture isolates.

Bacteroides fragilis is an important anaerobic pathogen accounting for up to 10% of bacteremias in adult patients. Enterotoxin producing B. fragilis (ETBF) strains have been identified as enteric pathogens of children and adults. In order to further characterize the B. fragilis pathogenicity island (BfPAI) and using PCR assays for bft- and mpII-metalloprotease genes, we determined the frequency of B. fragilis strains with pattern I (containing the BfPAI and its flanking region), pattern II (lacking both the BfPAI and the flanking region), and pattern III (lacking the BfPAI but containing the flanking region) in 63 blood culture isolates. The results were compared to 197 B. fragilis isolates from different clinical sources. We found 19% of blood culture isolates were pattern I (ETBF), 43% were pattern II (NTBF) and 38% were pattern III (NTBF). Comparatively, B. fragilis isolates from other clinical sources were 10% pattern I, 47% pattern II and 43% pattern III. This suggests that the pathogenicity island and the flanking elements may be general virulence factors of B. fragilis.

[Wound botulism in heroin addicts in Germany]. / Wundbotulismus bei Heroinabhängigen in Deutschland.

HISTORY AND ADMISSION FINDINGS: 5 heroin addicts (aged 31-44 years; 1 female, 4 men) presented with a history of blurred vision and diplopia followed by dysarthria. 3 of the patients also developed respiratory failure requiring long-term ventilatory support. Physical examination revealed cranial nerve deficits and abscesses at injection sites in 3 of them. DIAGNOSIS: In 4 patients wound botulism was diagnosed on the basis of symptoms, course of the illness and response to specific treatment. Clostridium botulinum was grown from wound swab in one patient. TREATMENT AND COURSE: Two of the patients, having been injected with antitoxin immediately after admission, were discharged almost symptom-free after only a few days. Adjuvant antibiotics and, in 3 patients, surgical débridement of the abscesses were needed. CONCLUSIONS: Progressive cranial nerve pareses in addicts who inject drugs intravenously or intramuscularly should raise the suspicion of wound botulism and require hospitalization. While indirect demonstration of toxin supports the diagnosis, false-negative results are common.

[Appropriate microbiological diagnostics]. / Sinnvolle Mikrobiologische Diagnostik.

In the industrialized world the threat of infectious diseases is mainly due to nosocomial infections and multi-resistant agents. In this context, microbiological evaluations have not only a benefit for the individual patient, but also allow to evaluate the local epidemiologic situation. However, quality and benefits are often compromised by incorrect specimen collection. This review attempts to summarize diagnostic procedures, collection and transport of appropriate specimens and relevant causative agents for prominent clinical manifestations of infectious diseases.