Influence of postmortem time on the outcome of blood cultures among cadaveric tissue donors

Tissue banks provide tissues of human cadaver donors for transplantation. The maximal time limit for tissue retrieval has been set at 24 h postmortem. This study aimed at evaluating the evidence for this limit from a microbiological point of view. The delay of growth in postmortem blood cultures, the identification of the species isolated and clinical/environmental factors were investigated among 100 potential tissue donors. No significant difference was found in the rate of donors with grown blood cultures within (25/65=38%) compared with after (24/65=37%) 24 h of death. Coagulase-negative staphylococci and gastro-intestinal microorganisms were isolated within and after 24 h of death. Two factors—antimicrobial therapy and “delay before body cooling”—were significantly inversely related with donors’ blood culture results. From a microbiological point of view, there is no evidence for avoiding tissue retrieval among donors after 24 h of death. Part of this study has been presented at the 14th Annual Scientific Meeting of the British Association of Tissue Banks (BATB), BioCity, Nottingham, UK, April 2006. The abstract of the oral presentation was published in the journal of Cell and Tissue Banking 2007;8:63–76.     

Evaluation of methods for the construction of collagenous scaffolds with a radial pore structure for tissue engineering

Type I collagen is used widely as a biomaterial. The structure of collagenous biomaterials, including pore sizes and general architecture, can be varied by a number of techniques. In this study, we developed a method to construct flat fibrillar type I collagen scaffolds, 6 cm in diameter and with a radially orientated pore structure, by the use of directional freezing. Different methodologies were tested, the optimal one being freezing of a collagen suspension inside-out, using a centrally positioned liquid nitrogen-cooled tube. Pore sizes could be varied by the use of different tube materials. Use of aluminium tubes resulted in radial scaffolds with a pore size of 20-30 μm, whereas use of stainless steel produced radial scaffolds with 70-100 μm pore sizes. Brass- and copper-based tubes produced scaffolds with less homogeneous radial pores, pore sizes being 90-100 and 50-80 μm, respectively. Fibreglass tubes gave even less uniformity (pore size 100-150 μm). Scaffolds were free of cracks, except in case of aluminium. Scaffolds with a radial inner structure may be especially suitable for tissue engineering of organs with a radial scaffold structure, such as the diaphragm.     

Typing of <Emphasis Type="Italic">Pneumocystis jirovecii</Emphasis> by multilocus sequencing: evidence of outbreak?

Different reports of Pneumocystis jirovecii pneumonia (PcP) outbreaks on oncology and transplant units suggest the possibility of a person-to-person transmission. Based on these reports, we searched retrospectively for possible PcP clusters in UZ Leuven in 2013. A movement and transmission map was established for all patients (n?=?21) with a positive PcP PCR on BAL fluid. BAL fluid samples from all patients with a positive PCR on the mitochondrial large subunit mRNA of P. jirovecii and possible cross exposure were typed with multilocus sequence typing (MLST). Five patients with a positive PcP PCR could have contact with another PcP patient. Another five patients with a weak positive PcP PCR on BAL fluid during the same period were also included. Based on the MLST typing of the BAL samples of these ten patients, there was no evidence of a PcP outbreak in UZ Leuven in 2013. MLST has proven to be a useful tool in genotyping and outbreak detection. From this case series, it could be concluded that current infection control precautions for P. jirovecii are appropriate in UZ Leuven. However, there is need for an international Pneumocystis database and more clarity in the geographic distribution of different P. jirovecii genotypes.     

Microbiological diagnostics of bloodstream infections in Europe—an ESGBIES survey

ObjectivesHigh-quality diagnosis of bloodstream infections (BSI) is important for successful patient management. As knowledge on current practices of microbiological BSI diagnostics is limited, this project aimed to assess its current state in European microbiological laboratories.MethodsWe performed an online questionnaire-based cross-sectional survey comprising 34 questions on practices of microbiological BSI diagnostics. The ESCMID Study Group for Bloodstream Infections, Endocarditis and Sepsis (ESGBIES) was the primary platform to engage national coordinators who recruited laboratories within their countries.ResultsResponses were received from 209 laboratories in 25 European countries. Although 32.5% (68/209) of laboratories only used the classical processing of positive blood cultures (BC), two-thirds applied rapid technologies. Of laboratories that provided data, 42.2% (78/185) were able to start incubating BC in automated BC incubators around-the-clock, and only 13% (25/192) had established a 24-h service to start immediate processing of positive BC. Only 4.7% (9/190) of laboratories validated and transmitted the results of identification and antimicrobial susceptibility testing (AST) of BC pathogens to clinicians 24 h/day. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry from briefly incubated sub-cultures on solid media was the most commonly used approach to rapid pathogen identification from positive BC, and direct disc diffusion was the most common rapid AST method from positive BC.ConclusionsLaboratories have started to implement novel technologies for rapid identification and AST for positive BC. However, progress is severely compromised by limited operating hours such that current practice of BC diagnostics in Europe complies only partly with the requirements for optimal BSI management.     

Transfusion-induced serum sickness

BACKGROUND: Transfusion-induced serum sickness reactions are rarely reported in the literature. The Type III hypersensitivity reaction to heterologous proteins involves deposition of complement and immune complexes in small vessel walls resulting in a leukocytoclastic vasculitis. A case of a multiply transfused patient with several episodes of serum sickness reactions is presented.
CASE REPORT: A 61-year-old man with myelodysplastic syndrome type refractory anemia presented with fever, rash, and polyarthralgia 5 days after transfusion of red blood cells (RBCs). By transfusing plasma-free "washed" RBCs, similar serum sickness reactions were avoided.
RESULTS: Laboratory investigation showed an increase of serum creatinine, hematuria, and proteinuria. Levels of circulating immune complexes immunoglobulin G and immunoglobulin M were increased. Hypocomplementemia could not be demonstrated. Histopathologic examination of the skin showed leukocytoclastic vasculitis, compatible with serum sickness.
CONCLUSION: The importance of early recognition of transfusion-induced serum sickness reactions is emphasized, because this can reduce unnecessary morbidity from this unusual complication of transfusion. To prevent this type of transfusion reaction, patients who experienced serum sickness–like reactions after transfusion should only receive plasma-free washed RBCs.     

Antimicrobial susceptibility of coagulase-negative staphylococci on tissue allografts and isolates from orthopedic patients.

Allograft infection occurs at a rate not different from that of similar procedures with large allografts or sterilized prosthetic devices and is usually caused by coagulase-negative staphylococci (CNS). CNS are feared for their limited antimicrobial susceptibility. We aimed at investigating this risk. CNS were isolated from 260 of 1461 allograft tissue grafts and compared with 384 consecutive clinical isolates from a general orthopedic population (258 patients). The CNS were identified and examined for their susceptibility to nine antibiotics used in routine practice. Staphylococcus epidermidis was the most commonly identified (35%) and the most resistant species of the allograft isolates. Comparing the overall antibiotic susceptibility patterns, clinical pathogens were significantly more resistant to six of the nine antibiotics (p < 0.01), namely penicillin, oxacillin, erythromycin, clindamycin, ofloxacin, and gentamicin. In conclusion, massive allograft infection is a well-known life-threatening surgical risk. However, we did demonstrate that allograft-related in contrast to orthopedic clinics-related CNS, are susceptible to commonly used first and second line antibiotics.     

Performance of three generations of Xpert MRSA in routine practice: approaching the aim?

The aim of this study was to evaluate retrospectively the performance of the Xpert MRSA assay in routine practice and its current use in the intensive care unit (ICU) setting of our hospital, since a pre-emptive isolation strategy has been applied. A total of 6473 patients were routinely screened with ESwab for methicillin-resistant Staphylococcus aureus (MRSA) using three generations of rapid real-time polymerase chain reaction (PCR) (Cepheid GeneXpert) over three consecutive periods of time. Performance was evaluated using broth enrichment culture as the reference method. Our results show that the last generation of Xpert MRSA (NxG) assay is more specific (99.2% vs. 97.9%) but not more sensitive (77.8% vs. 86.9%) than the third generation. Considering the low prevalence of MRSA in our hospital, we obtained an overall low positive predictive value. In conclusion, it remains difficult to abandon the reference method in routine practice considering the possible implications of an erroneous MRSA result in the ICU.     

‘Bedside assessment’ of acute hantavirus infections and their possible classification into the spectrum of haemophagocytic syndromes

Hantavirus infections, recently renamed ‘hantavirus fever’ (HTVF), belong to the most common but also most underestimated zoonoses in the world. A small number of reports described the so-called ‘lipid paradox’ in HTVF, i.e. the striking contrast between a very low serum total cholesterol and/or high-density lipoprotein cholesterol (HDLc), and a paradoxical concomitant hypertriglyceridaemia. In a prospective study, with patients being their own control after illness, we wanted to verify if this quick and easy ‘bedside test’ was robust enough to warrant a preliminary diagnosis of acute kidney injury (AKI) caused by HTVF. The study cohort consisted of 58 Belgian cases (mean age 44 years), admitted with varying degrees of AKI and of thrombocytopaenia, both characteristic for presumptive HTVF. All cases were sero-confirmed as having acute HTVF. At or shortly after hospital admission, a significant (p?<?0.001) decrease of total cholesterol and HDLc was found in comparison with normalised levels in the same cohort, quantified a few days after spontaneous AKI recovery. Conversely, fasting triglyceride levels during HTVF infection were significantly (p?<?0.001) higher during illness than after recovery. This ‘lipid paradox’ was most outspoken in severe HTVF cases, often accompanying, or even predicting, major kidney or lung complications. Thus, this ‘bedside assessment’ seems to hold even promise for presumptive diagnosis of more severe so-called ‘hantavirus cardio-pulmonary syndrome’ (HCPS) cases, mostly described hitherto in the New World. In more severe AKI cases, the mean total cholesterol was significantly lower (p?=?0.02) than in milder cases, i.e. cases with peak serum creatinine levels of?<?1.5 mg/dL. Thrombocytopaenia, generally accepted as the severity index in HTVF, appeared, moreover, significantly correlated with serum levels of total cholesterol (R?=?0.52, p?<?0.001) and with serum levels of HDLc (R?=?0.45, p?<?0.01). A link with the novel clinical entity of haemophagocytic syndromes, also characterised by manifest hypertriglyceridaemia, is discussed.     

Bacteriostasis testing on allograft tissue inoculated in Wilkins-Chalgren broth

Tissue banks culture tissue specimens to confirm the absence of viable micro-organisms after decontamination with antibiotics. It is possible that antibiotic residues attached to decontaminated tissue are introduced into enrichment culture media. These could have an inhibitory effect on the culture results and generate false-negative results. Our aim was to detect bacteriostasis in Wilkins-Chalgren broth inoculated with bone and tendon remnants. These remnants had been soaked in a solution containing gentamicin as part of the tissue-processing procedure. We used the United States Pharmacopeia method for bacteriostasis testing with gentamicin-susceptible Pseudomonas aeruginosa American Type Culture Collection (ATCC) 15442, Staphylococcus aureus ATCC 6538, Bacillus subtilis ATCC 6633 as test strains, and gentamicin-resistant Candida albicans ATCC 90029 as control. The residual gentamicin concentration in the broths was determined and gentamicin-soaked tissue was placed on Mueller-Hinton agar inoculated with a staphylococcal suspension. Bacteriostasis was present in 53-75% of the reference test strains. Tendon remnants had a significantly higher rate of bacteriostasis (85%) than bone remnants (28%). Broths inoculated with tendon remnants had the highest residual gentamicin concentrations.     

The oro-dental phenotype in Prader-Willi syndrome: a survey of 15 patients.

BACKGROUND: Prader-Willi syndrome (PWS) is a rare disorder caused by genetic defects in certain regions of chromosome 15q11-13. It is characterized by severe neonatal hypotonia and feeding problems, childhood-onset hyperphagia and obesity, short stature, facial dysmorphy, hypogonadism, learning and behavioural difficulties, and dental abnormalities. AIM: To describe the oro-dental phenotypic spectrum of patients with PWS. DESIGN: Fifteen PWS patients (3-35 years of age) being followed at the Centre for Human Genetics of the University Hospital of Leuven were examined at the dental clinic of the same institution. Medical information collected included age at diagnosis, body mass index (BMI) and level of cognitive functioning. Oral, clinical and radiological evaluations were performed. Caries experience (cavitation level), dental erosion and salivary flow rates were assessed. RESULTS: The 15 patients had dmft/DMFT scores ranging from 0 to 28, while nine were cavity-free. Those with severe caries experience also presented advanced dental erosion. BMI ranged from 16 to 42.6. There was no association between BMI and caries experience or erosive tooth wear. The PWS patients in our survey presented with a more favourable oral health status than those in previous studies. This might be due to early diet management or better oral hygiene during childhood or both.