Umbilical Cord Blood Cultures Research Articles

PROTAC-mediated vimentin degradation promotes terminal erythroid differentiation of pluripotent stem cells

BackgroundHuman pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs), can undergo erythroid differentiation, offering a potentially invaluable resource for generating large quantities of erythroid cells. However, the majority of erythrocytes derived from hPSCs fail to enucleate compared with those derived from cord blood progenitors, with an unknown molecular basis for this difference. The expression of vimentin (VIM) is retained in erythroid cells differentiated from hPSCs but is absent in mature erythrocytes. Further exploration is required to ascertain whether VIM plays a critical role in enucleation and to elucidate the underlying mechanisms.MethodsIn this study, we established a hESC line with reversible vimentin degradation (dTAG-VIM-H9) using the proteolysis-targeting chimera (PROTAC) platform. Various time-course studies, including erythropoiesis from CD34+ human umbilical cord blood and three-dimensional (3D) organoid culture from hESCs, morphological analysis, quantitative real-time PCR (qRT-PCR), western blotting, flow cytometry, karyotyping, cytospin, Benzidine-Giemsa staining, immunofluorescence assay, and high-speed cell imaging analysis, were conducted to examine and compare the characteristics of hESCs and those with vimentin degradation, as well as their differentiated erythroid cells.ResultsVimentin expression diminished during normal erythropoiesis in CD34+ cord blood cells, whereas it persisted in erythroid cells differentiated from hESC. Depletion of vimentin using the degradation tag (dTAG) system promotes erythroid enucleation in dTAG-VIM-H9 cells. Nuclear polarization of erythroblasts is elevated by elimination of vimentin.ConclusionsVIM disappear during the normal maturation of erythroid cells, whereas they are retained in erythroid cells differentiated from hPSCs. We found that retention of vimentin during erythropoiesis impairs erythroid enucleation from hPSCs. Using the PROTAC platform, we validated that vimentin degradation by dTAG accelerates the enucleation rate in dTAG-VIM-H9 cells by enhancing nuclear polarization.Graphical

Promise and pitfalls of umbilical cord blood culture for neonatal early-onset sepsis.

Promise and pitfalls of umbilical cord blood culture for neonatal early-onset sepsis.

Umbilical cord blood culture for early onset sepsis in preterm infants.

Umbilical cord blood culture (UCBC) may have a diagnostic utility for early onset sepsis (EOS) detection in preterm infants. It may prevent sampling the newborn and collect a higher volume of blood for pathogenic identification. Retrospective analysis at a tertiary care center in Canada of preterm infants ≤ 34 0/7 weeks' gestation with UCBC taken at birth. Of 505 admitted infants, 195 had UCBC. 170 UCBCs were negative; 44 of these had also negative neonatal blood culture (NBC). No infants with negative UCBC showed EOS symptoms in the first week of life. 25 UCBCs were positive: 18 were contaminants (all with negative NBC) and 7 were confirmed as EOS. 18 infants with UCBC contaminants remained asymptomatic. 7 EOS cases were identified, with varying bacteriological profiles; 5 displayed sepsis symptoms while 2 were asymptomatic. Risk of EOS increased with prolonged rupture of membranes. UCBC effectively detected EOS establishing it as a method with possibly better diagnostic performance than NBC in high-risk neonates. Further studies are needed to improve UCBC technique and lower contamination rates. Umbilical cord blood culture has a higher bacterial identification rate than peripheral venous blood culture for the early identification of early-onset sepsis in preterm infants. Umbilical cord blood cultures that showed no growth were reliable predictors of not developing early onset sepsis. Umbilical cord blood culture should be considered as part of the evaluation for early onset sepsis in the preterm infants.

Umbilical Cord Blood Culture as an Aid to the Diagnosis of Early Onset Neonatal Sepsis: A Cross-sectional Study

Introduction: Neonatal sepsis is one of the leading causes of neonatal morbidity and mortality globally, accounting for an estimated neonatal mortality rate of 23.9 per 1000 live births. Due to overlapping signs and symptoms, a specific diagnosis of sepsis poses a diagnostic challenge. Blood collected from a peripheral vein for sepsis screening and blood culture remains the gold standard for diagnosing neonatal sepsis. The umbilical cord is still not routinely used as a site for collecting blood for sepsis screening and blood culture. Aim: To determine the diagnostic efficacy of Umbilical Cord Blood Culture (UCBC) compared to Peripheral Venous Blood Culture (PVBC) in Early Onset Neonatal Sepsis (EONS). Materials and Methods: This cross-sectional observational study was conducted at the Neonatology Unit, Department of Paediatrics, Assam Medical College and Hospital (AMCH), Dibrugarh, Assam, India, involving 110 neonates with two or more risk factors for EONS over a one-year period (August 2021-July 2022). Umbilical cord blood and peripheral venous blood were collected and cultured, and the neonates were monitored throughout their hospital stay. Statistical significance was determined using the Chi-square test for categorical variables and the t-test for continuous variables (with a p-value <0.05 considered statistically significant). The validity of UCBC for diagnosing early neonatal sepsis was assessed based on sensitivity, specificity, Positive Predictive Value (PPV), and Negative Predictive Value (NPV). Results: The mean gestational age was 34.95±3.314 weeks and mean birth weight was 2.08±0.790 grams. Of the 110 high- risk neonates, sepsis screening was positive in 67 (61%), while UCBC and PVBC were positive in 19 (17.3%) and 10 (9.09%), respectively. Acinetobacter was the most common organism found in both cultures. The sensitivity and specificity of sepsis screening were 100% and 47.25% compared to UCBC and 90% and 42% compared to PVBC. In comparison to PVBC, UCBC demonstrated a sensitivity and specificity of 70% and 88%, with a diagnostic accuracy of 86.36%. Conclusion: The UCBC exhibits good diagnostic accuracy for diagnosing EONS and can be utilised due to it being a painless and technically less challenging method of blood sampling, with high sensitivity and specificity.

Umbilical Cord Blood IgA Levels and Bacterial Profile in Preterm Neonates Born with Maternal Risk Factors for Early-Onset Neonatal Sepsis.

To investigate the IgA levels and bacterial profile in umbilical cord blood (UCB) samples of mothers with risk factors compared to those without risk factors; and to understand the link between UCB culture positivity and neonatal outcomes [early-onset sepsis (EOS) or death within 7 d of life]. This is a pilot prospective case-control study. Mothers with preterm deliveries (gestational age <34 wk) were enrolled in two groups- Cases: Those with antenatal risk factors (prolonged duration of rupture of membranes of ≥24h or chorioamnionitis) and controls: Those without these two risk factors. Serum IgA levels was assayed and microbiological culture was tested in UCB samples. 16S sequencing to determine the UCB microbiome was performed in a subset of samples (n = 15). Neonates were followed-up for the occurrence of EOS or death until 7 d of life. Forty-nine mothers as cases and 50 mothers as controls were consecutively enrolled. No significant difference was observed in the IgA levels (60.5 vs. 58.1mg/L; p = 0.71), neonatal blood culture positivity (4.1% vs. 8.0%; p = 0.41) and UCB culture positivity (30.6% vs. 26.0%; p = 0.61) in the two groups.No difference was observed between the groups in occurrence of EOS or death within 7 d of life. Proteobacteria, Firmicutes and Actinobacteria were the most abundant phyla. Serratia, Bifidobacterium, Collinsella, Meganomas and Blautia being the most common genera. Cord blood IgA concentration could not differentiate the neonates at-risk of infection due to its presence in both the groups.

ROLE OF CORD BLOOD CRP AND CULTURES IN DIAGNOSIS OF NEONATAL SEPSIS

Background: Sepsis has killed about 30–40% of low-income neonates, often caused by maternal-fetal microorganisms. PROM affects 3% of pregnancies, aside from chorioamnionitis, untreated UTI, etc. Many studies have predicted neonatal sepsis by detecting inflammatory markers in venous blood 12–24 hours after birth. Some studies have tested CBC, CRP, and blood culture in babies with PROM and other risk factors to predict early-onset neonatal sepsis by drawing umbilical cord blood. Hence, we performed this study to determine the role of CRP and blood cultures from umbilical cord blood to determine early-onset sepsis. Methods: This was a prospective observational study performed in Sharda university, Greater Noida. 122 neonates within 72 hours of birth born via full term vaginal delivery, after 37 completed 37 weeks of gestation. The neonates were then divided in the two groups- neonates with likely sepsis or no sepsis based on the symptoms. Following this, umbilical cord blood was drawn at 24 hours and 72 hours. Results: we observed that 32.47% of the neonates in the likely sepsis group had confirmed evidence, while 3.4% in the no sepsis group we found to have evidence of sepsis. The differences was statistically significant (p&lt; 0.05). CRP was higher in umbilical cord blood and venous blood in likely sepsisneonates, in comparison to than blood culture yields and total WBC. Compared to the non-infected control group, the differences were statistically significant (P value 0.05). CRP demonstrated 100% sensitivity, specificity, PPV, and NPV in cord blood and 100%, 77.1%, 27.3%, and 100% in venous blood at 24 hours. Conclusion: Neonates with the higher risk of sepsis had higher readings in umbilical cord blood at birth and newborn venous blood at 24 hrs., but not at 72 hours, Also,negative CRP readings better excluded infection. Early-onset sepsis cannot be diagnosed by umbilical cord blood cultures.

Comparative microbial analysis and assessment of paired umbilical cord blood culture and peripheral venous blood culture in neonates with high risk factors to detect early onset neonatal sepsis: A study from tertiary care hospital of central India

Background: Neonatal sepsis is a prime cause of neonatal morbidity and mortality. Peripheral venous blood (PVB) culture is a gold standard for the diagnosis of neonatal sepsis. Low sensitivity of blood culture in newborn is due to small volume of blood sample and antibiotics given before sampling. Aims and Objectives: The study aims to compare and analyze the microbial spectrum in the Umbilical cord blood (UCB) culture with PVB culture and evaluation of both for detection of early onset neonatal sepsis (EONS) in high risk neonates and to use appropriate microorganism specific antibiotic for treating the sepsis. Materials and Methods: About 100 inborn neonates with two or more risk factors for EONS, chosen by sequential sampling method were included in this prospective analytical study. Blood samples were collected from umbilical cord and peripheral vein for culture. Sepsis screen was done to substantiate the diagnosis of neonatal sepsis. Results: Out of 100 neonates, 21 belongs to sepsis; 14 to probable sepsis; 65 to no sepsis. Among these 27 had UCB culture &amp; 24 had PVB culture +ve. Organisms grown on culture were Klebsiella(m/c isolate), Staphylococcus aureus, Escherichia coli, Acinetobacter, Coagulase-ve Staphylococcus (blood culture contaminant), Pseudomonas and Enterococcus. Conclusion: UCB culture is simple and convenient method for the diagnosis of neonatal sepsis as compared to PVB culture with Sensitivity of 66% and specificity of 93%. Organisms grown are comparable to PVB culture sample (Klebsiella - most common isolate).

Utility of Umbilical Cord Blood Culture in the Diagnosis of Early Onset Sepsis: A Cross-sectional Study

Introduction: Umbilical cord is the first source of blood from the neonate. The clinical signs associated with sepsis are frequently non specific and subtle in the neonates making the diagnosis of infection difficult. Umbilical cord blood does not involve pain infliction, avoids iatrogenic blood loss and procedural complications. Aim: To evaluate the utility of Umbilical Cord Blood Culture (UCBC) in the diagnosis of Early Onset Sepsis (EOS). Materials and Methods: The present study was a crosssectional study carried out in neonatology unit of Department of Paediatrics, Rural Medical College, Loni, Maharashtra, India, for the period of mid-March 2021 to mid-September 2021. Neonates delivered in Rural Medical College to the mothers having the risk factors for EOS were included in this study. Informed consent was taken prior to start of study. Thus, 68 samples were collected and studied. UCBC was collected with all aseptic precautions immediately after the umbilical cord was cut after the birth of the baby, venous blood sample was collected within one hour of birth. The data was presented as count and percentages. Results: Majority were female neonates 40 (58.8%). On analysing maternal risk factors it was seen 8.8% had previous low birth weight and 13.2% had Rh negative status. Analysis for presence of risk factors for sepsis majority 79.4% were multiple times examined per vaginally, followed by 36.8% had foul smelling liquor, 11.8% had febrile illness and 5.8% had birth asphyxia. On umbilical cord culture and sensitivity, most common microorganism identified was Staphylococcus aureus 8 (11.8%), followed by 8.8% Pseudomonas species. So, present study shows that 35.3% (24 cases) had positive culture reports using utility of UCBC in 68 patients. Conclusion: Study concludes that umbilical cord blood sampling and culture can be used as a tool for diagnosing bacterial sepsis in neonates especially the high-risk neonates.

Eliminating Contamination in Umbilical Cord Blood Culture Sampling for Early-Onset Neonatal Sepsis.

Introduction: Despite the advantages of umbilical cord blood culture (UCBC) use for diagnosis of early onset sepsis (EOS), contamination rates have deterred neonatologists from its widespread use. We aimed to implement UCBC collection in a level III neonatal intensive care unit (NICU) and apply quality improvement (QI) methods to reduce contamination in the diagnosis of early onset sepsis.Methods: Single-center implementation study utilizing quality improvement methodology to achieve 0% contamination rate in UCBC samples using the Plan-Do-Study-Act (PDSA) model for improvement. UCBC was obtained in conjunction with peripheral blood cultures (PBC) in neonates admitted to the NICU due to maternal chorioamnionitis. Maternal and neonatal characteristics between clinical sepsis and asymptomatic groups were compared. Process, outcome, and balancing measures were monitored.Results: Eighty-two UCBC samples were collected in addition to peripheral blood culture from neonates admitted due to maternal chorioamnionitis. Ten (12%) neonates had a diagnosis of clinical sepsis. All PBCs were negative and 5 UCBCs were positive in the study period. After 2 PDSA cycles, there was special cause variation with improvement in the percent of contaminated samples from 7.3 to 0%. There was no change in antibiotic duration among asymptomatic neonates.Conclusions: Implementation of UCBC for the diagnosis of EOS in term infants is feasible and contamination can be minimized with the implementation of a core team of trained providers and a proper sterile technique without increasing antibiotic duration.

Use of umbilical cord blood culture in the diagnosis of early onset neonatal sepsis among high risk mothers

Background: Early onset neonatal sepsis (EONS) is one of the important causes of morbidity and mortality in neonates. Its early diagnosis and prompt treatment is essential and any delay in the diagnosis can have serious consequences including neonatal death. Blood culture is the gold standard test for diagnosis of neonatal sepsis. Umbilical cord blood culture (UCBC) is a painless procedure and technically less challenging. We conducted this study to evaluate use of UCBC for the diagnosis of EONS and compared it with the results of peripheral venous blood culture (PVBC) reports. Aims and Objectives: The aim of the study was to evaluate UCBC for the diagnosis of EONS and compared it with the results of PVBC reports. Materials and Methods: This was a hospital-based prospective cohort study consisting of 100 neonates who were at risk of EONS. The study was conducted in the Department of Pediatrics Sikkim Manipal Institute of Medical Sciences Gangtok between January 2018 and December 2019. Neonates found to be at risk of development of EONS were included in this study on the basis of a predefined inclusion and exclusion criteria. Immediately after birth blood samples were collected from both umbilical cord and peripheral vein and were sent to bacteriology lab. Sensitivity, specificity, positive predictive value, and negative predictive value of both the samples were analyzed. Results: Out of 100 neonates in 32 (32%) EONS could be confirmed with positive sepsis screening results and/or demonstration of organisms on blood culture. Among the 32 neonates with EONS, 17 were found to be premature. The mean gestational age of newborns with EONS was found to be 35.2 weeks. The umbilical blood culture was found to have sensitivity and specificity of 100% and 74.4%, respectively, whereas peripheral vein blood culture was found to have sensitivity and specificity of 77.7% and 72.5%, respectively. The most common organism grown in our study was Escherichia coli. Conclusion: UCBC is painless and technically less challenging method of blood sampling. It has been found to have a higher sensitivity as well specificity for the diagnosis of EONS as compared to peripheral venous blood sample.

Umbilical cord blood culture in neonatal early-onset sepsis: a systematic review and meta-analysis.

Peripheral blood culture (PBC) is considered the gold standard for diagnosis of neonatal early-onset sepsis (EOS), but its diagnostic value can be questioned. We aimed to systematically asses the diagnostic test accuracy (DTA) of umbilical cord blood culture (UCBC) for EOS. A systematic literature search was performed in PubMed, Embase, Web of Science, and the Cochrane Library. Studies performing UCBC for the diagnosis of EOS were included. A total of 1908 articles were screened of which 17 were included. Incidences of positive PBC and UCBC were low in all studies. There was a large heterogeneity in the consistency between positive PBC and UCBC outcomes. PBC had a pooled sensitivity of 20.4% (95% CI 0.0-40.9) and specificity of 100.0% (95% CI 100.0-100.0) compared to 42.6% (95% CI 12.7-72.4%) and 97.8% (95% CI 93.1-100.0) of UCBC for clinical EOS, defined as clinical sepsis regardless of PBC outcomes. This systematic review shows that, compared to PBC, UCBC has higher sensitivity and comparable specificity for clinical EOS and might be considered as diagnostic test for EOS. Due to the limited number of studies, low incidences of EOS cases, and the imperfect reference standards for EOS, results should be interpreted cautiously. This is the first systematic review and meta-analysis investigating the diagnostic test accuracy of umbilical cord blood culture for neonatal early-onset sepsis. Peripheral blood culture is considered the gold standard for diagnosis of neonatal early-onset sepsis, but its value for this specific diagnosis can be questioned. Umbilical cord blood culture has higher sensitivity and comparable specificity for diagnosis of neonatal early-onset sepsis compared to peripheral blood culture, circumventing the risk for iatrogenic anemia and consequently might be used as a diagnostic tool for early-onset sepsis. Quality of evidence was regarded as low due to imperfect diagnostic methods of neonatal early-onset sepsis.

A PROSPECTIVE COMPARATIVE STUDY OF UMBILICAL CORD BLOOD CULTURE VERSUS PERIPHERAL VENOUS BLOOD CULTURE IN DIAGNOSIS OF EARLY ONSET NEONATAL SEPSIS IN PRETERM NEONATES IN NEONATAL INTENSIVE CARE UNIT OF TERTIARY CARE HOSPITAL

Background Neonatal sepsis is major cause of neonatal morbidity and mortality. Blood culture and sensitivity is gold standard for the diagnosis of neonatal sepsis. Low sensitivity of blood culture especially in newborn is due to small volume of blood sample collected from neonates and antibiotics given before sampling. Study Methodology A prospective, analytical, cross sectional study where comparison of umbilical cord blood culture and peripheral venous blood culture was done in 100 inborn neonates . Sensitivity and specificity, positive and negative predictive values were calculated. P value was calculated, Chi Square test was applied and association was quantified. Results Higher sensitivity(81.0%) and accuracy(87%) for predicting disease outcome of patients by UCBC method against PVBC method conclude that UCBC can be used as reliable and alternate tool to predict final outcome. Conclusion UCBC can be useful diagnostic test for EONS. Adequate amount of blood is obtained by umbilical cord method which can be difficult to obtain in a low birth weight new born. Inadequate samples of PVBC can lead to a false negative report. Umbilical cord blood is obtained from placental end of umbilical cord, there are less chances of introducing iatrogenic infection unlike routine culture technique where chances of false positive and risk of introducing iatrogenic infection is high. If umbilical cord blood is taken at birth, normal appearing newborn could be followed up in postnatal wards and discharged if reports are normal. If umbilical cord blood is collected at birth and culture report comes positive,we can prevent sequelae of sepsis by providing antibiotics for fixed duration and for units practising empirical antibiotics , antibiotics can be discontinued if report is negative.

Limited Mitochondrial Activity Coupled With Strong Expression of CD34, CD90 and EPCR Determines the Functional Fitness of ex vivo Expanded Human Hematopoietic Stem Cells

Ex vivo expansion strategies of human hematopoietic stem cell (HSC) grafts with suboptimal stem cell dose have emerged as promising strategies for improving outcomes of HSC transplantation in patients with hematological malignancies. While exposure of HSCs to ex vivo cultures expands the number of phenotypically identifiable HSCs, it frequently alters the transcriptomic and metabolic profiles, therefore, compromising their long-term (LT) hematopoietic reconstitution capacity. Within the heterogeneous pool of expanded HSCs, the precise phenotypic, transcriptomic and metabolic profile and thus, the identity of HSCs that confer LT repopulation potential remains poorly described. Utilizing valproic acid (VPA) in ex vivo cultures of umbilical cord blood (UCB)-CD34+ cells, we demonstrate that expanded HSCs phenotypically marked by expression of the stem cell markers CD34, CD90 and EPCR (CD201) are highly enriched for LT-HSCs. Furthermore, we report that low mitochondrial membrane potential, and, hence, mitochondrial activity distinguishes LT-HSCs within the expanded pool of phenotypically defined HSCs. Remarkably, such reduced mitochondrial activity is restricted to cells with the highest expression levels of CD34, CD90 and EPCR phenotypic markers. Together, our findings reveal that high expression of CD34, CD90 and EPCR in conjunction with low mitochondrial activity is critical for identification of functional LT-HSCs generated within ex vivo expansion cultures.

Umbilical Cord Blood Culture in General in the Diagnosis of Sageis Newborns

Worldwide neonatal sepsis is among the most frequent causes of neonatal death. Various studies have tried to establish the relationship between prevalence of neonatal septicemia risk factors and bacteriological profiling, low birth weight, prematurity, etc. Current study was aimed to compare early onset of neonatal sepsis (EONS) among primigravida and multigravida mothers using umbilical cord blood (UCB) and peripheral venous blood (PVB) samples. It was also aimed to establish the utilization of umbilical cord blood culture (UCBC) in comparison to peripheral venous blood culture (PVBC) in identifying EONS. In present study the blood samples were collected from high risk neonates for the clinical blood culture and screening. Among the 75 neonates in the study, 24 (32.0%) were observed to have sepsis screen positive. Study of high risk neonates umbilical cord blood culture (UCBC) positivity was 17.3% while Peripheral Venous blood culture positivity was 5.3%. Moreover, in this study all risk factors like Prematurity, Low birth weights, Premature rupture of membrane, and birth asphyxia were significantly (p&lt;0.05) associated with UCBC growth/positivity. Low birth weight (86%) was mostly reported in the high risk neonates with other associated sepsis factors. Similarly maternal fever and prolonged rupture of membrane was highly significantly (p&lt;0.01) associated with UCBC positivity. Gram negative bacterias were more commonly found, such as Pseudomonas (5.3%), followed by E. coli (4%), and Klebsiella (2.7%) and gram positive Streptococcus sp. (2.7%), etc. From our analysis it can be said that the UCBC has strong diagnostic outcomes as compared to the PVBC for etiological evaluation of bacterial sepsis in neonates at high risk.

Umbilical Cord Blood Culture in Diagnosis of Early Onset Neonatal Sepsis.

To evaluate the use of umbilical cord blood culture (UCBC) as compared to neonatal blood culture (NBC) in the diagnosis of early onset neonatal sepsis (EONS). This hospital based prospective observational study was conducted in the Department of Pediatrics and Department of Obstetrics & Gynaecology at a tertiary care centre of North India from 2017 through 2018. A total of 80 newborns with presence of two or more risk factors for sepsis were included in the study. Blood culture samples were collected from the umbilical cord at delivery and from the neonate within 1h of birth. UCBC was positive in 17 (21.2%) neonates and NBC was positive in 15 (18.7%) neonates. Out of these positive cases, 10 (45.4%) neonates had similar bacteriological profile in both UCBC and NBC which included Burkholderia cepacia, Acinetobacter species, Enterobacter cloacae and Coagulase-negative Staphylococcus. UCBC had a sensitivity of 66.7%, a specificity of 89.2%, a positive predictive value 58.8% and a negative predictive value 92.1% for the diagnosis of EONS in high risk neonates, considering NBC as the gold standard test. UCBC is a reliable alternative to NBC for early etiological diagnosis of EONS in high risk neonates. Additional blood sampling from the newborn is suggested if there is any clinical sign of sepsis.

Detection of bla and bla OXA-48 genes in Gram-negative isolates from neonatal sepsis in central of Iran

The aim of this study was to determine the prevalence of neonatal sepsis with a focus on antibiotic resistance and the frequency of the bla CTX-M-15 and bla OXA-48 genes in Gram-negative isolates. A total of 108 Umbilical Cord Blood (UCB) and 153 peripheral blood samples were cultured via BACTEC from May 2017 to June 2018. The bacterial isolates were identified using phenotypic and genotypic analyses. The antibiotic susceptibility profile of the isolates was determined by disk diffusion. PCR was used to determine the frequency of β-lactamase genes. Among the 153 infants, 21 (13.7%) proved positive for sepsis. Escherichia coli, Staphylococcus epidermidis and Klebsiella pneumoniae were the most frequent isolates in the peripheral blood cultures. E. coli and Stenotrophomonas maltophilia were isolated from two UCB cultures. The highest resistance among the Gram-positive strains was to cefixime, ceftriaxone, cefotaxime and clindamycin. In the Gram-negative bacteria the highest rates of resistance were to ampicillin (91.7%). The frequency of bla OXA-48 and bla CTX-M-15 genes was 25% and 50%, respectively. The high antibiotic resistance among the isolates reveals the importance of monitoring antibiotic consumption and improving control standards in the health care system, especially in neonatal wards.

Modulation of S. epidermidis-induced innate immune responses in neonatal whole blood.

Coagulase-negative staphylococci (CoNS) such as Staphylococcus epidermidis are highly prevalent pathogens for sepsis in neonates. The interaction between host, environment and pathogenic factors of S. epidermidis are still poorly understood. Our objective was to address the role of several pathogenic factors of S. epidermidis on neonatal cytokine responses and to characterize the influence of three immunomodulatory drugs. We performed an ex-vivo model of S. epidermidis sepsis by assessment of blood cytokine production in neonatal whole blood stimulation assays (ELISA). S. epidermidis strains with different characteristics were added as full pathogen to umbilical cord blood cultures and the influence of indomethacin, ibuprofen and furosemide on neonatal immune response to S. epidermidis was evaluated (Flow cytometry). Stimulation with S. epidermidis sepsis strains induced higher IL-6 and IL-10 expression than stimulation with colonization strains. Biofilm formation in clinical isolates was associated with increased IL-10 but not IL-6 levels. In contrast, stimulation with mutant strains for biofilm formation and extracellular virulence factors had no major effect on cytokine expression. Notably, addition of ibuprofen or indomethacin to S. epidermidis inoculated whole blood resulted in mildly increased expression of TNF-α but not IL-6, while frusemide decreased the production of pro-inflammatory cytokines, i.e. IL-6 and IL-8. The virulence of sepsis strains is coherent with increased cytokine production in our whole-blood in-vitro sepsis model. Biofilm formation and expression of extracellular virulence factors had no major influence on readouts in our setting. It is important to acknowledge that several drugs used in neonatal care have immunomodulatory potential.

Reduction of Phlebotomy-Related Blood Loss in Neonates Through Use of Umbilical Cord Blood Cultures

Reduction of Phlebotomy-Related Blood Loss in Neonates Through Use of Umbilical Cord Blood Cultures

Umbilical cord blood culture versus peripheral venous blood culture in early onset neonatal sepsis

Background: Neonatal sepsis is a major cause of neonatal morbidity and mortality. Blood culture and sensitivity is gold standard for the diagnosis of neonatal sepsis. Low sensitivity of blood culture especially in newborn is due to small volume of blood sample collected from neonates and antibiotics given before sampling. The aim was to evaluate the use of umbilical cord blood culture in the diagnosis of early onset neonatal sepsis as compared to Peripheral vein blood culture. Methods: Eighty neonates with two or more risk factors for early onset neonatal sepsis were included in the study. Blood samples were collected from umbilical cord and peripheral vein for culture. Sepsis screen was done to corroborate the diagnosis of neonatal sepsis. Results: Sepsis screen was positive in twenty three babies. Among these four had grown organism on Umbilical cord blood culture only. While two babies had both positive, Umbilical cord bloods culture and peripheral vein blood culture. Organisms grown on culture were E.Coli, Pseudomonas, Klebsiella and Acinetobacter.Conclusions: Umbilical cord blood culture is simple and convenient method for the diagnosis of early onset neonatal sepsis compared to peripheral venous blood culture. Organisms grown are comparable to peripheral venous blood culture sample.

Microbial Contamination and Evaluation of Inoculum Volume in Umbilical Cord Blood Culture

Background: Microbial screening tests of umbilical cord blood (UCB) are essential for stem cell transplantation. We analyzed the microbial contamination rate and distribution of isolated microorganisms over 10 years of samples from the MEDIPOST Cord Blood Bank. In addition, we studied the influence of inoculum volume microorganism culture and compared the yield and speed of microorganism detection. Methods: Microbial screening tests were performed using a manual method, which includes using an inoculum of 2 mL of plasma, a byproduct of UCB processing from pediatric culture bottles. When positive blood culture was detected, each set was once again inoculated with 2 mL and 4 mL of plasma. Results: From 2004 to 2013, a total of 133,610 UCB units were screened, of which 1,311 (0.9%) tested positive for contamination. The most frequently identified microorganism was Escherichia coli (34.6%), followed by Bacillus spp. (12.8%), Enterococcus faecalis (5.3%) and Klebsiella pneumoniae (4.4%). The total yield rate increased by 0.2% over this time period, although the yield rate of Bacillus spp. increased by 8.3%. Conclusion: The results of this study could be used in many ways with both domestic and international data regarding cord blood contamination. Also, other microbiology laboratories using culture conditions similar to ours could refer this study when preparing guidelines. Finally, by detecting low levels of bacteria, we have contributed to cord blood safety. (Ann Clin Microbiol 2016;19:1-6)