The Procurement Team as a Factor of Bone Allograft Contamination

We studied the effect of the procurement team on the risk of contamination in 270 large bone allografts retrieved from 53 non-living donors under strictly aseptic conditions.The overall contamination rate was 8.1%. When the procurement team was constituted by three or less members the contamination rate was 5.6%, while if there were four or more members the rate was 12.9%; this difference was significant in the statistical analysis.We conclude that a procurement team constituted by three or less trained members is a determinant factor in decreasing the bacterial contamination rate of bone allografts.     

Isolation of bacteria of the family enterobacteriaceae from plant tissues

Bacteria of the family Enterobacteriaceae were isolated from the tissues of a number of wild and cultivated plants. All the cultures isolated had a broad spectrum of resistance to antibiotics and were highly adhesive to human erythrocytes. The studies conducted indicate the possibility of concentration of microorganisms pathogenic for humans in plant tissues.     

The Incidence of Hepatitis C Virus Positive Serological Test Results Among Cornea Donors

Purpose. We have noticed that the incidence of positive serological tests of hepatitis C virus (HCV) among cornea donors in our eye bank is higher than expected. The purpose of this study was to determine the incidence of these positive findings and identify the contributory risk factors. Methods. All corneas procured between June 1993 and June 1997 were included in this retrospective study. In all cases a routine work-up, including serological testing of donors' sera and evaluation of the procured corneas, was performed prior to corneal transplantation. Donors found to be positive for HCV antibodies were compared to our general population of corneal donors, with respect to the demographic, serological and ophthalmological data obtained during the transplantation work-up. Results. All 851 corneas procured from 438 donors were included. Antibodies to HCV were found in 29 donors (6.6%). Following positive test results, 57 corneas (6.7%) were discarded. The time from donors' death to cornea procurement was significantly longer among HCV-positive patients than in the general donor population (12.3 vs. 9.3h, p<0.0003). No other differences were detected between the two groups. Conclusion. Delay in harvesting of donated corneas may give rise to false positive HCV-antibodies test results, which may be partially responsible for the high rate of positive findings among cornea donors, with consequent tissue wastage.     

Isolation of Bacteria of the Family <Emphasis Type="Italic">Enterobacteriaceae</Emphasis> from Plant Tissues

Bacteria of the family Enterobacteriaceae were isolated from the tissues of a number of wild and cultivated plants. All the cultures isolated had a broad spectrum of resistance to antibiotics and were highly adhesive to human erythrocytes. The studies conducted indicate the possibility of a concentration of microorganisms pathogenic for humans in plant tissues.     

Analysis of potential factors affecting allografts contamination at retrieval

The microbiological contamination of retrieved tissues has become a very important topic and it is a critical aspect in the safety of allografts, especially from multi-tissue donors whose tissues are frequently contaminated as a consequence of retrieval. We analysed a total of 10,107 tissues, 8178 musculoskeletal and 1929 cardiovascular tissues, retrieved from 978 multi-tissue donors. Of these, 159 heart-beating donors (HBD) were also organ donors, while the remaining 819 non-heart-beating donors (NHBD) were tissue donors only. A multivariate logistic model was used to determine the factors affecting contamination risk during retrieval. In the model, the dependent variable was the presence/absence of contamination while the covariates included were: gender, type of donor, age of donor, cause of death, previous skin donation, cadaver time, number of people attending the retrieval, number of tissues retrieved. Moreover, a second log-linear model was used to determine the number of strains isolated per tissue. Tissue contamination was statistically correlated with gender, type of donor, cadaver time, number of people attending the retrieval and season. In conclusion, to minimize the risk of bacterial contamination, aseptic techniques should be used at retrieval, with the number of retrieval team members kept to a minimum. In addition, cadaver time should be as short as possible and the donor should be refrigerated within a few hours after death.     

Metal Removal from Contaminated Soils Through Bioleaching with Oxidizing Bacteria and Rhamnolipid Biosurfactants

The use of surfactants as a method for solubilization and removal of heavy metal contamination from soil has been reported before. Biosurfactants produced by some microorganisms are able to modify the surface of various metals and aggregate on interphases favoring the metal separation process from contaminated environments. We evaluated the feasibility of enhancing the removal of metal ions from mineral waste/contaminated soils using alternate cycles of treatment with rhamnolipid biosurfactants and bioleaching with a mixed bacterial culture of Acidithiobacillus thiooxidans and Acidithiobacillus ferrooxidans. Bioleaching alone removed 50% Zn and 19% Fe. When rhamnolipids were used at low concentration (0.4 mg/mL), 11% Fe and 25% Zn were removed, while at 1 mg/mL 19% Fe and 52% Zn removal were achieved. When using a cyclic treatment combining bioleaching and biosurfactants, metal removal reached up to 36% for Fe and 63% to 70% for Zn.     

Analysis of predisposing factors for contamination of bone and tendon allografts

Bone and tissue allografts are widely used in transplantation. The increasing demand for safe allografts must be met, while minimizing disease transmission. We analysed the incidence and potential risk factors of allograft contamination and the effectiveness of disinfection, by reviewing 22 years of tissue bank activity and 474 donor procurements. We also compared different disinfection procedures used over the 22 years. The overall contamination rate was 10.1%. Risk factors were related to the donor or procurement method. Immediate culture at the tissue recovery site diminished the rate of false positives by reducing later sample manipulation. High-virulence allograft contamination was mainly related to donor factors, while low-virulence contamination was related to procurement methods. Analysis of donor-related risk factors showed no statistical differences for age, sex, or cause of death. An intensive care unit stay was associated with less contamination with high-virulence microbes. Procurement in a setting other than an operating theatre was associated with higher contamination rate. Team experience reduced contamination. Pelvic and tendon allografts were most frequently contaminated. Proper disinfection considerably reduced the contamination rate to 3.6%. We conclude that procurement must be performed under aseptic conditions, with short delays, and by trained personnel. Grafts should be disinfected and packed as soon as possible.     

Skin donors and human skin allografts: evaluation of an 11-year practice and discard in a referral tissue bank

The Saint Louis hospital tissue bank provides skin allografts to pediatric and adult burn units in the Paris area. The aim of this study was to analyze our activity during the last 11 years focusing on the reasons for skin discard. Skin is procured solely from the back of the body, which is divided into 10 zones that are harvested and processed separately. This retrospective study included all skin donors harvested between June 2002 and June 2013, representing a total of 336 donors and 2770 zones. The donors were multiorgan heart-beating donors in 91 % of cases (n = 307). The main reason for discarding harvested skin was microbial contamination, detected in 99 donors (29 %). Most contaminants were of low pathogenicity. Other reasons for discard included positive serologic tests for 2 donors [17 zones (0.61 %)], unsuitable physical skin characteristics for 3 zones (0.11 %), the donor’s medical history for 53 zones (1.91 %), and technical issues with processing or distribution for 61 zones (2.2 %). In our experience, microbial contamination continues to be the main reason for discarding potential skin allografts. However, discards are limited by separate harvesting and processing of multiple zones in each donor.     

Molecular testing of <Emphasis Type="Italic">Klebsiella pneumoniae</Emphasis> contaminating tissue allografts recovered from deceased donors

Microbiological screening of tissue allografts is crucial to prevent the transmission of bacterial and fungal infections to transplant recipients. Klebsiella was the most prevalent and resistant contaminating microorganism observed in our setting in the Iranian Tissue Bank. This study was conducted to determine the presence of extended-spectrum β-lactamase (ESBL) genes, antimicrobial resistance patterns of Klebsiella pneumoniae isolates, and their clonal relationships in allograft materials. K. pneumoniae contaminating bone and other tissue allografts recovered from deceased donors were identified and ESBL isolates were detected using a phenotypic confirmatory method. Antimicrobial susceptibility testing was carried out using the disk diffusion method. Distribution of ESBL genes and molecular typing were performed using polymerase chain reaction (PCR) and Repetitive-element (rep-PCR) methods. Of 3828 donated tissues, 51 (1.3%) were found contaminated by K. pneumoniae isolates. Compared to tissue allografts from brain-dead, heart-beating tissue donors, allografts from donors with circulatory cessation were associated with a higher risk of K. pneumoniae contamination [odds ratio (OR), 1.2 (CI 95% 0.9–2.3) (P value < 0.001)]. Half of the isolates produced ESBL, and the rate of susceptibility to cephalosporins was 51%. Among isolates, 22 (43.1%) harbored CTX-M, 31 (60.8%) SHV, and 9 (17.6%) harbored TEM types. The rep-dendrogram indicated that clones having identical or related strains with a similar antibiotype were isolated in the same period. This study provides evidence that a single clone of K. pneumoniae contaminated tissue allografts recovered from many different donors. A single clone found on tissues from several donors suggests contamination of tissues from a single source such as the tissue recovery process and environment. Genomic DNA testing and clonality of contaminating bacteria using molecular methods can focus the epidemiologic investigation on the tissue allograft recovery process including a search for contamination of the tissue recovery room environment, recovery staff, recovery equipment, reagents, solutions and supplies.     

Assessment of cryopreserved donor skin viability: the experience of the regional tissue bank of Siena

Skin allografts from cadaver donors are an important resource for treating extensive burns, slow-healing wounds and chronic ulcers. A high level of cell viability of cryopreserved allografts is often required, especially in burn surgery, in Italy. Thus, we aimed to determine which conditions enable procurement of highly viable skin in our Regional Skin Bank of Siena. For this purpose, we assessed cell viability of cryopreserved skin allografts procured between 2011 and 2013 from 127 consecutive skin donors, before and after freezing (at day 15, 180, and 365). For each skin donor, we collected data concerning clinical history (age, sex, smoking, phototype, dyslipidemia, diabetes, cause of death), donation process (multi-tissue or multi-organ) and timing of skin procurement (assessment of intervals such as death-harvesting, harvesting-banking, death-banking). All these variables were analysed in the whole case study (127 donors) and in different groups (e.g. multi-organ donors, non refrigerated multi-tissue donors, refrigerated multi-tissue donors) for correlations with cell viability. Our results indicated that cryopreserved skin allografts with higher cell viability were obtained from female, non smoker, heartbeating donors died of cerebral haemorrhage, and were harvested within 2 h of aortic clamping and banked within 12 h of harvesting (13–14 h from clamping). Age, cause of death and dyslipidaemia or diabetes did not appear to influence cell viability. To maintain acceptable cell viability, our skin bank needs to reduce the time interval between harvesting and banking, especially for refrigerated donors.     

重症监护室烧伤感染患者脓毒症的病原菌分布及影响因素研究

目的:探讨重症监护室(ICU)烧伤感染患者脓毒症的发生情况及病原菌分布,并分析其危险因素,为防治ICU烧伤感染患者脓毒症提供理论依据。方法:收集我院2016年7月-2019年7月期间的86例ICU烧伤感染患者的临床资料,依据是否发生脓毒症,将其分为脓毒症组和非脓毒症组,比较两组的基本资料等情况,分析ICU烧伤感染患者脓毒症的发生率及病原菌分布,并分析ICU烧伤感染患者脓毒症发生的影响因素。结果:86例ICU烧伤感染患者中有33例发生脓毒症,发生率为38.37%;ICU烧伤感染患者脓毒症病原菌主要是革兰阴性杆菌(占54.55%),其次是革兰阳性球菌(占42.42%);单因素结果分析发现,烧伤至入院时间、合并糖尿病、吸入性损伤、首次手术时间以及烧伤面积与ICU烧伤感染患者脓毒症的发生有关(P<0.05);多因素Logistic回归分析发现,首次手术时间>2d、TBSA>50%是ICU烧伤感染患者脓毒症发生的独立危险因素(P<0.05)。结论:ICU烧伤感染患者易发生脓毒症,首次手术时间>2d以及TBSA>50%是其发生的独立危险因素,临床应采取针对性预防措施,以降低ICU烧伤感染患者脓毒症发生率。     

ICU下呼吸道多重耐药菌医院感染的病原学临床特征及易感因素分析

摘要 目的：研究ICU下呼吸道多重耐药菌医院感染的病原学临床特征及易感因素。方法：选择2020年1月到2022年12月于我院ICU住院治疗的216例下呼吸道感染者,按照是否发生多重耐药菌感染分为研究组113例,对照组103例。分析两组患者感染相关因素的数量分布情况,通过Logistic回归分析多重耐药菌医院感染的危险因素。采用全自动细菌鉴定仪对菌种进行鉴定,采用K-B纸片法进行药敏试验,并分析多重耐药菌感染的病原学分布及对常用抗菌药物的耐药性。结果：（1）与对照组相比,研究组患者感染相关因素的分布率更高;（2）住院时间>3个月、使用糖皮质激素治疗、应用机械通气治疗、其他细菌感染、血红蛋白含量<100 g/L、抗菌药物使用时间>15 d、抗菌药物使用种类>4种、使用免疫抑制剂是ICU下呼吸道多重耐药菌感染的危险因素;（3）113例研究组共培养出细菌菌株93株,其中革兰氏阴性菌52株（55.91%）,革兰氏阳性菌25株（26.88%）,革兰氏阴性菌中较多的是铜绿假单胞菌（22株）、鲍曼不动杆菌（13株）、肺炎克雷伯菌（12株）;革兰氏阳性菌中最多的是肺炎链球菌（11株）和金黄色葡萄球菌（11株）;（4）耐药情况：铜绿假单胞菌对莫西沙星耐药率较低（15.83%）,肺炎克雷伯菌对亚胺培南耐药率较低（17.56%）,鲍曼不动杆菌对头孢哌酮/舒巴坦耐药率较低（16.37%）,金黄色葡萄球菌、肺炎链球菌对万古霉素无耐药性。结论：住院时间>3个月、使用糖皮质激素治疗、应用机械通气治疗、其他细菌感染、血红蛋白含量<100 g/L、抗菌药物使用时间>15 d、抗菌药物使用种类>4种、使用免疫抑制剂是多重耐药感染的独立危险因素。本院ICU下呼吸道感染以革兰氏阴性杆菌为主,应根据病原菌选择耐药性低的药物,并针对危险因素采取有效措施。     

Bacterial infection transmitted by human tissue allograft transplantation

Bacterial contamination of tissue allografts obtained from cadaveric donors has been a serious cause of morbidity and mortality in recipients. Recent cases of fatal and nonfatal bacterial infections in recipients of contaminated articular cartilage (distal femur) and tendon allografts have called attention to the importance of avoiding tissue donors suspected of carrying infectious disease, of not processing donated tissue carrying virulent bacteria, the occurrence of falsely negative final sterility tests, and the need to sterilize tissues. These cases demonstrated that contamination can arise from an infected donor, during tissue removal from cadaveric donors, from the processing environment, and from contaminated supplies and reagents used during processing. Final sterility testing can be unreliable, especially when antibiotics remain on tissues. There is an increasing need for control of microbial contamination in tissue banks, and sterilization of tissue allografts should be recommended whenever possible.     

The appropriateness of swab cultures for the release of human allograft tissue

Surgeries utilizing human allograft tissues have increased dramatically in recent years. With this increase has come a greater reliance on the use of swab culturing to assess allograft tissues for microbial contamination prior to distribution. In contrast to the typical industrial microbiological uses for swabs, the tissue banking industry has relied on swab cultures as a sterility release method for allograft tissues. It has been reported in the literature that swabs have limitations, both in sensitivity and reproducibility, so their suitability as a final sterility release method was evaluated in this study. Two different swab-culturing systems were evaluated (COPAN, EZ Culturette) using human allograft tissues spiked with low levels of multiple bacterial and fungal microorganisms. The average microbial recoveries for all challenge microorganisms for each tissue type and each swab system were calculated. Percent recoveries for each challenge microorganism were also calculated and reported. The results indicated that both swab systems exhibited low and highly variable recoveries from the seeded allograft tissues. Further analysis indicated there was no statistical difference (∝=0.05) between the two swab systems. It is the recommendation of the authors that swab culturing not be used to assess relatively low levels of microbial contamination on allografts. Instead, alternative validated microbial detection methods with improved sensitivity and reproducibility should be employed and validated for this critical task.     

Endoscopic versus transcranial procurement of allograft tympano-ossicular systems: a prospective double-blind randomized controlled audit

Allograft tympano-ossicular systems (ATOS) have proven their use over many decades in tympanoplasty and reconstruction after resection of cholesteatoma. The transcranial bone plug technique has been used in the past 50 years to procure en bloc ATOS (tympanic membrane with malleus, incus and stapes attached). Recently, our group reported the feasibility of the endoscopic procurement technique. The aim of this study was to assess whether clinical outcome is equivalent in ATOS acquired by using the endoscopic procurement technique compared to ATOS acquired by using the transcranial technique. A double-blind randomized controlled audit was performed in a tertiary referral center in patients that underwent allograft tympanoplasty because of chronic otitis media with and without cholesteatoma. Allograft epithelialisation was evaluated at the short-term postoperative visit by microscopic examination. Failures were reported if reperforation was observed. Fifty patients underwent allograft tympanoplasty: 34 received endoscopically procured ATOS and 16 received transcranially procured ATOS. One failed case was observed, in the endoscopic procurement group. We did not observe a statistically significant difference between the two groups in failure rate. This study demonstrates equivalence of the clinical outcome of allograft tympanoplasty using either endoscopic or transcranial procured ATOS and therefore indicates that the endoscopic technique can be considered the new standard procurement technique. Especially because the endoscopic procurement technique has several advantages compared to the former transcranial procurement technique: it avoids risk of prion transmission and it is faster while lacking any noticeable incision.     

Bioleaching of Heavy Metals from Textile Sludge by Indigenous Sulfur-and-Iron-Oxidizing Microorganisms Using Elemental Sulfur and Ferrous Sulfate as Energy Sources: A Comparative Study

The present study was undertaken to investigate the potential of enriched indigenous sulfur-and-iron-oxidizing microorganisms in the bioleaching of Cu, Ni, Zn and Fe from textile sludges by using elemental sulfur and ferrous sulfate (FS), respectively, as an energy source under batch conditions. The experiments were performed with three different textile sludges (S1, S2 and S3) at initial neutral pH of the sludges procured from different parts of the country i.e., UP, Haryana and Punjab. The three sludges used were not only procured from different parts of the country but also differ in physiochemical characteristics. The extent of heavy metals solubilization in each sludge was found to be different using sulfur- and iron-oxidizing microorganisms. The results of the study indicate that sulfur-oxidizing microorganisms were found more efficient in the bioleaching process, irrespective of any sludge. The use of sulfur-oxidizing microorganisms led to higher solubilization of heavy metals and after 7 days of bioleaching about 84–96% Cu, 64–78% Ni, 81–92% Zn and 74–88% Fe were removed compared to 62–73% Cu, 62–66% Ni, 74–78% Zn and 70–78% Fe using iron-oxidizing microorganisms. This study had shown the feasibility of applying the bioleaching process to textile sludge contaminated with heavy metals. The results of the present study indicate that the bioleached sludge would be safer for land application.     

Permanence can be Defended

In donation after the circulatory‐respiratory determination of death (DCDD), the dead donor rule requires that the donor be dead before organ procurement can proceed. Under the relevant limb of the Uniform Determination of Death Act 1981 (USA), a person is dead when the cessation of circulatory‐respiratory function is ‘irreversible’. Critics of current practice in DCDD have argued that the donor is not dead at the time organs are procured, and so the procurement of organs from these donors violates the dead donor rule. We offer a new argument here in defence of current DCDD practice, and, in particular, of the interpretation of the requirement of ‘irreversibility’ as permanence.     

Skin allograft procurement and transplantation in Mashhad,Iran: Are burn patients’ needs being met?

Only a single donor’s tissue may save or improve lives of one hundred patients. Unfortunately, low governmental and media support of tissue procurement and transplantation programs is a worldwide problem. Loss of an effective tissue procurement program in many countries like Iran, may lead to loss of many thousands valuable tissues each year. To evaluate the rate of skin donation in Mashhad in comparison to other organs and tissues, we extracted the data related to tissue and organ procurement in Mashhad from 2001. Then we evaluated the annual skin allograft needs in the Burn Department of Imam Reza Hospital as the only referral burn center in the northeast of Iran. Brain dead potential donation rate per million populations of Mashhad in the years 2007–2014 was about 33. The mean refusal rate was 51%. Of patients who have consent for donation, more than 86% have consent for skin donation. Skin allograft procured from 119 (35.5%) candidates. Average of skin retrieval per cadaveric was 1525 cm² with a gradual increase from 1400 cm² in the first year to 1800 cm² in the last year. The recipient to donor ratio was 1.14. It is estimated that about 1 cm² of skin allograft is needed for any cm² burnt body surface area. Considering more than 700 acute burn hospitalization in our burn unit, the patients need for skin allograft would be more than 3.5 million cm², annually. The annual amount of skin procurement in Mashhad has been currently about 20,000 cm². It shows that our patients demand is higher than supply. Skin procurement and transplantation is a simple procedure which can be as lifesaving as organ procurement and transplantation. But there isn’t any national organization to regulate tissue procurement, banking and transplantation. Governmental support of skin procurement and transplantation programs especially nonprofit programs may improve skin procurement rate and save more lives of severely burnt patients.     

Microbiological contamination in genetically modified animals and proposals for a microbiological test standard for national universities in Japan.

The Biosafety Committee of the Japanese Association of Laboratory Animal Facilities of National Universities (JALAN) investigated recent episodes of microbiological contamination in genetically modified mice (GMM), and the countermeasures taken when the contaminated GMM were introduced into animal facilities, by questionnaires addressed to 53 animal facilities belonging to JALAN and serological tests. Although almost all of the contaminated GMM were accepted with conditions such as rederivation after or before reception and housing in designated rooms, contamination with a spectrum of microorganisms was demonstrated in GMM transferred domestically and from abroad. In serological tests, Mycoplasma pulmonis, mouse parvovirus, and mouse encephalomylitis virus were detected in GMM transferred from domestic facilities and from abroad. The present results of the questionnaires and serological tests suggest that GMM are highly and widely contaminated with microorganisms compared with mice from commercial breeders. Thus, we propose a microbiological requirement, including microbiological status--excellent, common, and minimum--as a guide for the transfer and procurement of mice and rats in Japan.