Antibiotic therapy in case of positive cultures of kidney transplant preservation fluid: a nationwide survey of prescribing practices.

Our survey aimed to describe current prescribing practices for perioperative antibiotic prophylaxis in French kidney transplant centers. We conducted a nationwide cross-sectional clinical vignette-based survey that we sent via email to hospital practitioners involved in perioperative management of kidney transplant patients (KTR). Nearly half of practitioners contacted (182/427, 42.6%) were respondents. A total of 167 getting enough kidney transplant activity were eligible for the survey. The response rate was 50.7% (68/134) among interns and 33.8% (99/293) among seniors. Positive perfusion fluids (PF) cultures for methicillin-susceptible Staphylococcus aureus were associated with antibiotic prescribing in 35% of cases, with no difference in prescribing in patients with diabetes, obesity, or delayed graft function. Antibiotic prescribing was most frequent with Pseudomonas aeruginosa (67%) and Klebsiella pneumoniae strains producing extended spectrum ß-lactamases (57%). About 77%, 16%, and 13% of respondents, respectively, reported the existence of local practice guidelines for surgical antibiotic prophylaxis, a standardized approach for antibiotic prescribing in case of positive kidney transplant PF cultures, and local practice guidelines for systematical antibiotic prophylaxis in the early post-transplant period. In France, antibiotic prophylaxis practices in the perioperative kidney transplant period are very heterogeneous. To prevent unnecessary prescribing and bacterial resistance, evidence-based practice guidelines should be developed.

Microbiological epidemiology of preservation fluids in transplanted kidney: a nationwide retrospective observational study.

OBJECTIVES: Kidney transplant recipients are at high-risk for donor-derived infections in the early post-transplant period. Transplant preservation fluid (PF) samples are collected for microbiological analysis. In case of positive PF cultures, the risk for the recipient is unknown and there is no consensus for prescribing prophylactic antibiotics. This nationwide observational study aimed to determine the epidemiology of bacterial and fungal agents in kidney transplant PF cultures and identify risk factors associated with positive PF cultures. METHODS: We performed a retrospective observational study on the following data collected from a national database between October 2015 and December 2016: characteristics of donor, recipient, transplantation, infection in donor and PF microbiological data. RESULTS: Of 4487 kidney transplant procedures, including 725 (16.2%, 725/4487) from living donors, 20.5% had positive PF cultures (living donors: 1.8%, 13/725; deceased donors: 24.1%, 907/3762). Polymicrobial contamination was found in 59.9% (485/810) of positive PF cultures. Coagulase-negative staphylococci (65.8%, 533/810) and Enterobacteriaceae (28.0%, 227/810) were the most common microorganisms. Factors associated with an increased risk of positive PF cultures in multivariable analysis were (for deceased-donor kidney transplants): intestinal perforation during procurement (OR 4.4, 95% CI 2.1-9.1), multiorgan procurement (OR 1.4, 95% CI 1.1-1.7) and en bloc transplantation (OR 2.5, 95% CI 1.3-4.9). Use of perfusion pump and donor antibiotic therapy were associated with a lower risk of positive PF cultures (OR 0.4, 95% CI 0.3-0.5 and OR 0.6, 95% CI 0.5-0.7, respectively). CONCLUSION: In conclusion, 24% of deceased-donor PF cultures were positive, and PF contamination during procurement seemed to be the major cause.

A two-centre validation study of sterility test of corneal storage media with elimination of interfering antimicrobials in compliance with the European Pharmacopoeia.

The purpose of this study was to validate the sterility test of corneal culture and deswelling/transport media using a device for removal of antimicrobials before incubation in BACTEC™ automated system in two Italian Eye Banks. Corneal culture medium, TISSUE-C, and deswelling/transport medium, CARRY-C, were inoculated with 10-100 cfu of six European Pharmacopoeia (EP) reference strains and either treated with medical device RESEP for removal of antimicrobials (RESEP+ group) or left untreated (RESEP- group) before injection into the BACTEC Plus bottles. The same steps were repeated in the absence of inocula with tryptone soy broth samples as negative controls, and the inocula were also directly injected in the BACTEC™ bottles as growth controls. All the samples were incubated in BACTEC™ automated system for 7 days, and the time to detection of microbial growth was recorded automatically. At both the Eye Banks, in the RESEP+ groups, microbial growth was detected in 100% of samples. In the RESEP- group, the method sensitivity ranged from 66.7 ± 21.1 to 88.9 ± 6.4% for TISSUE-C samples while for CARRY-C samples the method sensitivity ranged from 94.5 ± 5.1 to 100%. The method specificity corresponded to 100% for all the groups at both Eye Banks. This two-centre validation study showed that the use of RESEP increased the sensitivity of sterility test using BACTEC™ automated system up to 100% and, consequently, allowed validation of the method for sterility testing of corneal storage and deswelling/transport media according to the EP requirements. The test could not be validated without the use of RESEP.

Serologic Evaluation of Cornea Donors and Microbiologic Evaluation of Cornea Storage Media in an Eye Bank from Izmir, Turkey.

OBJECTIVES: Our objective was to evaluate the serologic positivity of cornea donors and microbiologic positivity of cornea storage media at the Ege University Tissue and Cornea Bank, Izmir, Turkey. MATERIALS AND METHODS: We retrospectively investigated the serologic blood sample and microbiological culture media analysis results of all cornea donors at Ege University Tissue and Cornea Bank between 2007 and 2015 with reference to age, sex, and cause of death of each donor. RESULTS: Mean age of the 955 deceased donors was 43.19 ± 15.89 years (range, 2-65 y). The mean postmortem time to blood sample removal and excision of the cornea tissue was 8.4 hours (range, 4-12 h). Serologic analyses showed that 855 donors (89.5%) were seronegative. The remaining donors were seropositive for hepatitis B (54 donors; 5.7%), human immunodeficiency (27 donors; 2.8%), hepatitis C (14 donors; 1.5%), and syphilis (5 donors; 0.5%) virus infections. Microbiologic analyses of the storage media were negative, with no microorganisms shown in 855 samples (89.5%). Candida species (32 donors; 3.4%), Escherichia coli (14 donors; 1.5%), Pseudomonas aeruginosa (11 donors; 1.2%), methicillin-resistant Staphylococcus aureus (11 donors; 1.2%), Enterobacter species (11 donors; 1.2%), Klebsiella pneumoniae (7 donors; 0.7%), Acinetobacter baumannii (6 donors; 0.6%), Proteus species (5 donors; 0.5%), and Corynebacterium species (3 donors; 0.3%) were the detected microorganisms in the infected storage media. CONCLUSIONS: False-positive serologic results among cornea donors were high. The incidence of false-positive results might be decreased by earlier blood removal from deceased donors and testing of all potential donors in intensive care units. Although rare, endophthalmitis after keratoplasty might be a devastating problem. In addition to serologic testing, microbiologic analyses of cornea storage media before transplant may be an effective way to prevent postoperative infectious complications.

MicroRNA profiles in graft preservation solution are predictive of ischemic-type biliary lesions after liver transplantation.

BACKGROUND & AIMS: Ischemic-type biliary lesions (ITBL) are the second most common cause of graft loss after liver transplantation. Though the exact pathophysiology of ITBL is unknown, bile duct injury during graft preservation is considered to be a major cause. Here we investigated whether the release of cholangiocyte-derived microRNAs (CDmiRs) during graft preservation is predictive of the development of ITBL after liver transplantation. METHODS: Graft preservation solutions (perfusates) and paired liver biopsies collected at the end of cold ischemia were analysed by RT-qPCR for CDmiR-30e, CDmiR-222, and CDmiR-296 and hepatocyte-derived miRNAs (HDmiRs) HDmiR-122 and HDmiR-148a. MicroRNAs in perfusates were evaluated on their stability by incubation and fractionation experiments. MicroRNA profiles in perfusates from grafts that developed ITBL (n=20) and grafts without biliary strictures (n=37) were compared. RESULTS: MicroRNAs in perfusates were proven to be stable and protected against degradation by interacting proteins. Ratios between HDmiRs/CDmiRs were significantly higher in perfusates obtained from grafts that developed ITBL (p<0.01) and were identified as an independent risk factor by multivariate analysis (p<0.01, HR: 6.89). The discriminative power of HDmiRs/CDmiRs in perfusates was validated by analysis of separate brain death- (DBD) and cardiac death donors (DBD; p ≤ 0.016) and was superior to expression in liver biopsies (C=0.77 in perfusates vs. C<0.50 in biopsies). CONCLUSIONS: This study demonstrates that differential release of CDmiRs during graft preservation is predictive of the development of ITBL after liver transplantation. This provides new evidence for the link between graft-related bile duct injury and the risk for later development of ITBL.

Positive cultures of organ preservation fluid predict postoperative infections in solid organ transplantation recipients.

OBJECTIVE: The significance of positive cultures of organ preservation fluid (OPF) in solid organ transplantation is not known. We sought to describe the microbiology and define the clinical impact of positive OPF cultures. DESIGN: Retrospective cohort study. SETTING: Tertiary care hospital. PATIENTS: A consecutive sample of all solid organ transplantations at our center between July 2006 and January 2009 was reviewed. A total of 331 allografts (185 kidneys, 104 livers, 31 pancreases, and 11 hearts) met the inclusion criterion of having OPF cultures taken from the transplanted allograft. METHODS: Organisms recovered from OPF were classified as high or low risk according to their virulence. Clinical outcomes were compared between recipients of organs with positive OPF cultures and recipients of organs with negative OPF cultures. RESULTS: OPF cultures were positive in 62.2% of allografts and yielded high-risk organisms in 17.8%. Normal skin flora constituted the majority of positive OPF cultures, while Enterobacteriaceae spp. and Staphylococcus aureus made up the majority of high-risk organisms. Recipients of allografts with positive OPF cultures developed more frequent bacterial infections, regardless of allograft type (relative risk, 2.39; 95% confidence interval [CI], 1.61-3.54). Moreover, isolation of a given organism in OPF samples was associated with the development of a clinical infection with the same organism, regardless of allograft type. CONCLUSIONS: Positive cultures of OPF are common events in solid organ transplantation, frequently involve high-risk organisms, and are associated with the development of postoperative clinical bacterial infections. Further study is required to determine the optimal strategies for their prevention and management.

Microbiological findings of culture-positive preservation fluid in liver transplantation.

Bacterial and fungal infections are the leading cause of mortality in liver transplant (LT) recipients. Few studies have examined the incidence of culture-positive preservation fluid (PF) and the outcome of related recipients. The aim of this study was to determine the incidence and the microbiologic findings of PF positive cultures, and to evaluate the impact on morbidity and mortality of LT recipients. A retrospective analysis of PF cultures performed after 477 LTs from cadaveric grafts between January 2001 and February 2008 was conducted. Forty-five (9.5%) PFs were found to be positive with 1 or 2 pathogens. The demographic profiles of recipients of PF with positive or negative cultures were similar. Enterobacteriaceae species were the most frequent organisms (n = 30), followed by Staphylococcus aureus (n = 5), coagulase-negative staphylococci (n = 5), enterococci (n = 4), and yeasts (n = 3). Mortality rate at 1 month was not significantly different in recipients with positive or sterile PF cultures (88.1% vs. 87.7%, respectively). The rate of bacteremia among LT recipients with positive or negative PF cultures was not statistically different. Systemic infections caused by the pathogen cultured from the PF occurred in 8 (18%) of the 45 recipients, including bacteremia (4/8) or intra-abdominal sepsis (5/8). Causative organisms were Enterobacteriaceae species (n = 5), Candida species (n = 2), and Enterococcus faecium (n = 1). Among the 8 patients who developed infection with the PF organism, 4 (50%) died in the intensive care unit (ICU) vs. an ICU mortality rate of 8% (3/37) in those who did not develop infection with the PF organism (P < 0.05). Infection occurred less frequently in recipients who received antimicrobial therapy with activity against the PF isolate than in those without appropriate treatment (41% vs. 3.8%, P < 0.005). Those who develop infection with organisms recovered from PF cultures appear to have high early mortality rates; therefore, appropriate antimicrobial therapy against organisms cultured from PF should be given.

Detection of viral DNA in kidney graft preservation and washing solutions is predictive of posttransplant infections in pediatric recipients.

BACKGROUND: In pediatric kidney transplant recipients, viral infections occur soon after transplant and may be transmitted from the graft. METHODS: This study of 75 pediatric kidney transplants investigated whether genome sequences of parvovirus B19, Epstein-Barr virus (EBV), human cytomegalovirus (HCMV), and BK polyomavirus (BKV) could be detected in kidney graft samples (graft biopsy samples and preservation and washing solutions) collected before implantation and whether their presence was a risk factor for infections in the recipient. RESULTS: B19 DNA was detected in approximately 30% of graft biopsy samples, preservation solutions, and washing solutions; EBV DNA was detected in approximately 20% of preservation and washing solutions but rarely in biopsy samples; and HCMV DNA and BKV DNA were rarely detected in graft biopsy samples. Seronegative recipients of B19 DNA-positive and EBV DNA-positive grafts had a significantly higher risk of infection during the early posttransplant period than did recipients of negative grafts. In particular, none of the B19-seronegative recipients of B19 DNA-negative grafts experienced infection soon after transplant, whereas most recipients of B19 DNA-positive grafts experienced infection within the first month after transplant. CONCLUSIONS: Molecular testing of donor grafts for viruses that infect circulating and resident cells in the graft-such as B19 in the kidney-could be useful (in association with donor/recipient serostatus) for identifying recipients at high risk for posttransplant infections.

Estudio comparativo de dos soluciones de preservación en la función inicial del trasplante bipulmonar en humanos / Comparative study of two preservation solutions in the initial function after bilateral human lung transplantation

Objetivo. Comparar la influencia de dos líquidosde preservación, Euro-Collins (EC) y Perfadex(P), en la función del injerto pulmonar en la faseinicial del trasplante pulmonar en humanos.Diseño. Estudio retrospectivo.Ámbito. Unidad de trasplante pulmonar de laUnidad de Cuidados Intensivos (UCI) de un hospitaluniversitario.Pacientes. Un total de 79 pacientes fueron sometidosa un trasplante bipulmonar. Los injertospulmonares fueron preservados con EC en 23casos y con P en 56 casos.Variables de interés. Se valoró la función pulmonaral ingreso en la UCI mediante el cocientede presión parcial de oxígeno/fracción inspiradade oxígeno (PaO2/FiO2). También se valoró lamortalidad a los 30 días, la disfunción del injerto,así como la estancia en la UCI y el tiempo deventilación mecánica.Resultados. El cociente PaO2/FiO2 fue significativamentemejor en el grupo P que en el ECtanto al ingreso (258 ± 116 frente a 176 ± 105, p <0,01) como a las 12 horas (313 ± 109 frente a 248± 116, p < 0,05) del ingreso en UCI. La incidenciade disfunción del injerto fue menor en el grupo Pque en el EC (p < 0,045). No hubo diferencias encuanto a la mortalidad a los 30 días, estancia enla UCI y el tiempo de ventilación mecánica entreambos grupos.Conclusión. La preservación del injerto pulmonarcon P como líquido de preservación, comparadocon EC, se asocia con una mejor funcióndel injerto en las fases iniciales del trasplante bipulmonary con un descenso en la incidencia dela disfunción del injerto

Objective. Compare the influence of two preservationliquids, Euro-Collins (EC) and Perfadex(P) in the pulmonary graft function in the initialphase of lung transplant in humans.Design. Retrospective study.Scope. Lung transplant unit of the ICU of auniversity hospital.Patients. A total of 79 patients were subjectedto a transplant of both lungs. The pulmonarygrafts were preserved with EC in 23 cases andwith P in 56 cases.Variables of interest. Pulmonary function wasassessed on admission in the intensive care unit(ICU) with the PaO2/FiO2 ratio. Mortality, graftdysfunction stay in ICU and time of mechanicalventilation were also assessed at 30 days. Results. The PaO2/FiO2 ratio was significantlygreater in the P group than in the EC both on admission(p < 0.006) and at 12 hours (p = 0.032) inthe ICU. Graft dysfunction incidence was less ingroup P than in EC (p < 0.045). There were no differencesin regards to mortality at 30 days, stayin ICU and time of mechanical ventilation betweenboth groups.Conclusion. Preservation of the pulmonarygraft with P as preservation liquid compared withEC is associated with better graft function in theinitial phases of transplant of both lungs andwith a decrease in the incidence of graft dysfunction

[Chemical content analysis of specimens of human head temporal portion commercially distributed for otological surgery lessons].

Specimens of the human head temporal portion are provided in the United States and distributed commercially for lessons in otological surgery via the internet. Many otologists have obtained and used these specimens in Japan. According to our chemical content analysis, these specimens were found to contain harmful substances as well as large molecule aldehydes and fatty acid methylesters, which are not or only rarely included in the human cadaveric specimens prepared in Japan. We discuss the suggested treatment of these imported specimens, how trial preparation of specimens are much better than the imported items, and the background of the rapid and wide distribution of the imported items in Japan.

Lung preservation.

Better understanding of the mechanisms of ischemia-reperfusion injury, improvement in the technique of lung preservation, and the recent introduction of a new preservation solution specifically developed for the lungs have helped to reduce the incidence of primary graft dysfunction after lung transplantation. Currently, the limitation in extending the ischemic time is more often related to the increasing use of non-ideal lung donors rather than to poor lung preservation. In this review, we have focused our attention on the experimental and clinical work performed to optimize the methods of lung preservation from the time of retrieval to the period of reperfusion after graft implantation.

Anionic polysaccharides. A class of substances with hepatoprotective and antiadhesive properties in rat liver preservation.

In liver preservation, the substitution of the anion Cl(-) by lactobionic acid (LB) prevents reperfusion edema and extends the preservation time for human livers. We studied the effect of compounds that are structurally related to lactobionic acid: anionic polycarbohydrates (sulfated anionic polysaccharide, SAP, and pentosan polysulfate, PPS) on liver function and leukocyte-endothelial cell interaction in isolated perfusion and liver transplant models. Rat livers, cold-stored (24 h) in a Cl(-) -containing control solution, became edematous during 1 h of reperfusion. Substitution of Cl(-) by either LB, SAP, or PPS decreased reperfusion edema in a Cl(-) concentration-dependent fashion. Reperfusion edema was abolished completely after preservation in 100 mM SAP solution or PPS solution. Also hepatic lactic dehydrogenase (LDH) and aspartate aminotransferase (ASAT) release was lowest after preservation in those solutions. After preservation in LB or anionic polycarbohydrate solutions, portal venous resistance was significantly higher than after preservation in Cl(-)-containing control solution. Capillary blood flow was 391 +/- 83 pl/s and 398 +/- 174 pl/s after preservation in SAP solution (SAPs) and PPSs, and 803 +/- 117 pl/s and 641 +/- 219 pl/s after preservation in LB or Cl(-)-containing control solution. The number of leukocytes sticking to the vascular wall was lower ( P < 0.05) after preservation in SAPs or PPSs (109 +/- 31 cells/mm(2) and 108 +/- 60 cells/mm(2), respectively), when compared with preservation in Cl(-)-containing control or LB solutions (429 +/- 63 cells/mm(2) and 277 +/- 59 cells/mm(2)). In rat liver preservation, anionic polysaccharides are antiedematous compounds, with a higher potency than LB and additional antiadhesive properties.

Água de coco como meio de cultura em conservante de córnea: estudo experimental em coelhos / Coconut water as culture medium in storage corneal medium: experimental study in rabbits

Objetivo: Verificar se a água de coco pode ser utilizada como meio de cultura na composição de conservante de córneas. Local: Laboratório de Cirurgia Experimental do Departamento de Cirurgia da Universidade Federal do Ceará (U.F.C.), Laboratório de Tecnologia de Sêmem de Caprinos do Departamento de medicina Veterinária da Universidade Estadual do Ceará (U.F.C.), Sociedade de Assistência aos Cegos e Centro de Oftalmologia do Ceará (OFTALMED). Método: Foram realizadas paquimetrias ultra-sônicas e microscopias espetaculares, em oito córneas de coelho, durante 14 dias de preservação em três meios: com água de coco, controle (sem água de coco) e com o meio (OPTISOL). Após 14 dias realizou-se o exame histopatológico das córneas. Foram aplicadas 2 gotas do conservante com água de coco em olhos de coelhos para pesquisar agressão ocular pelo conservante.

Residual ethanol content of donor sclera after storage in 95% ethanol and saline rinse of various durations.

PURPOSE: Some surgeons are wary of using alcohol-preserved sclera for allografts because they fear a toxic effect on surrounding tissue after placement. We set out to determine the amount of ethanol remaining in scleral allograft material after storage in 95% ethanol. METHODS: Sixty half scleras from 30 donors were preserved in 95% ethanol for an average of 31+/-14 days (range, 11 to 50 days). Rehydration was performed by soaking each half sclera in 4 ounces of balanced salt solution. Half scleras were randomly assigned to six groups of 10 each. Assays for ethanol were performed on the following groups: no balanced salt solution soak and balanced salt solution soak for 10 minutes, 20 minutes, 30 minutes, 40 minutes, and 50 minutes. Ethanol assay was performed by Headspace Gas Space Chromatography at ChemaTox Laboratory, Inc, Boulder, Colorado. RESULTS: The 10 half scleras without balanced salt solution soak had a mean ( SD) ethanol level of 175+/-14.1 mg per g of sclera. After 10 minutes of balanced salt solution s oak, the level decreased to 7.57+/-1.56 mg per g, then 3.77+/-3.02 mg per g at 20 minutes, 1.59+/-0.61 mg per g at 30 minutes, 1.07+/-0.30 mg per g at 40 minutes, and 0.96+/-0.26 mg per g at 50 minutes. Approximately 96% of the ethanol is leeched out of the half sclera by 10 minutes and 98% by 20 minutes. CONCLUSIONS: For sclera preserved in 95% ethanol, soaking in balanced salt solution for 20 minutes or longer leeches approximately 98% of the ethanol from the preserved donor sclera.