Microbiological profiles of tracheostomy patients: a single-center experience

Background: This study compared the prevalence of common microorganisms in obstructed and non-obstructed cases across the four quarters on the first post-tracheostomy year.Methods: A retrospective chart review of the microbiological profiles of all adult patients who underwent a tracheostomy was conducted between June 2015 and September 2019 at our hospital. Based on the tracheostomy indications, patients were allocated to obstructed or non-obstructed group. Any patient with at least one positive sample was followed up quarterly for a year. The first culture result obtained was recorded at least one month following the last antibiotic dose in each quarter.Results: Out of the 65 tracheal aspirate results obtained from 58 patients (mean age, 57.5±16.48 years), the most common procedure and indications were surgical tracheostomy (72.4%) and non-obstructed causes (74.1%), respectively. Moreover, 47.7% of the culture results indicated Pseudomonas aeruginosa, which showed significantly different proportions across the quarters (p=0.006). Among obstructed patients, P. aeruginosa was the most common (35%), followed by methicillin-resistant Staphylococcus aureus (MRSA; 23.5%).Conclusions: The most common post-tracheostomy microorganism was P. aeruginosa. MRSA showed a strong association with tracheostomy for obstructive indications.

IDENTIFICATION OF FORMALDEHYDE IN UNBRANDED WET NOODLES AT TRADITIONAL MARKETS OF TAMBUN SELATAN USING TEST KIT METHODS AND UV-VIS SPECTROPHOTOMETRY

Introduction: Wet noodles are flour-based foods that are widely consumed by the community because of their easy processing. The high water content in wet noodles makes the noodles not last long. In fact, there is still a preservative in wet noodles that is not used, namely formaldehyde. The purpose of this study was to determine whether there was formaldehyde in unbranded wet noodles in the Traditional Market of South Tambun District and to determine the level of formaldehyde in unbranded noodles in the Traditional Market of South Tambun District. Method: This research was conducted at the STIKes Mitra Keluarga laboratory. The type of research used is descriptive. There are 7 samples taken from 4 different markets. The sample was tested using a formaldehyde test kit and resulted in 6 positive formaldehyde samples. According to SNI, formaldehyde should not be in wet noodles. So that the results of the 6 samples did not meet the quality requirements of wet noodles. Results: Furthermore, the positive sample was determined using a UV-VIS spectrophotometer and Nash reagent. The lowest level of formaldehyde is 18.61 mg/L while the highest level is 738.45 mg/L. Conclusion: First, the regulation is not yet known by the producers because it is classified as traditional. Second, there is no good control from authorized institutions or officers. Third, the lack of guidance for producers

Detección de Pasteurella multocida, Mannhemia haemolytica, Histophilus somni y Mycoplasma bovis en pulmón de bovinos

In this study, it was aimed to determine of P. multocida, M. haemolytica, H. somni and M. bovis in macroscopically healthy cattle lungs by PCR. The study was carried out on 82 macroscopically healthy cattle lung. DNA extraction was performed to the lung samples. PCR was then performed using all specific primers. By molecular evaluation, positive results were achieved for P. multocida, M. haemolytica, H. somni and M. bovis in 4 (4.8 %), 4 (4.8 %), 6 (7.3 %) and 3 (3.6 %) of the samples, respectively. Mix infections were detected in five samples. Of the samples, two were positive for both P. multocida and M. haemolytica, two were positive for both M. haemolytica and H. somni and one was positive for both P. multocida and H. somni. However, a positive sample, which carried all of pathogens, was not detected. In conclusion, P. multocida, M. haemolytica, H. somni and M. bovis are the important opportunistic pathogens of respiratory tract in cattle and these pathogens have a major role during infections. But multifactorial nature of bovine respiratory disease and immune system affected the formation of the disease. Hence, firstly cattle’s immunity should be strengthened and other conditions should be kept under control.

Epidemiological characteristics and genetic diversity of norovirus infections among outpatient children with diarrhea under 5 years of age in Beijing, China, 2011–2018

Background Human noroviruses are the leading cause of sporadic cases and outbreaks of viral acute gastroenteritis in all age groups worldwide. Methods Epidemiological data and fecal specimens were collected between January 2011 and December 2018 from 4911 children < 5 years of age with diarrhea in three districts of Beijing. From 2011 to 2013, One-Step Reverse Transcription Polymerase Chain Reaction (RT-PCR) was used to detect noroviruses, and from January 2014 to December 2018, norovirus GI and GII were screened using duplex quantitative real-time RT-PCR (qRT-PCR). One-Step RT-PCR and RT-seminested PCR were performed to amplify the RNA-dependent polymerase and capsid genes of noroviruses in positive sample. Amplified products were sequenced directly; norovirus was typed using the online Norovirus Genotyping Tool v2.0 and phylogenetic analyses were conducted using MEGA-X. Results From 2011 to 2018, noroviruses were detected in 16.5% of specimens from children with diarrhea. The highest prevalence was observed in children aged 12 to 23 months (22.4%, 319/1421), followed by children aged 6 to 11 months (17.6%, 253/1441). The highest prevalence of norovirus infections occurred in autumn followed by winter, spring, and summer. From 2011 to 2018, the most prevalent dual types (genotype and polymerase type) were GII.4 Sydney[P31] (51.6%, 239/463), followed by GII.3[P12] (24.0%, 111/463), GII.4 2006b[P4 2006b] (7.3%, 34/463), GII.2[P16] (5.0%, 23/463), GII.17[P17] (2.6%, 12/463) and GII.6[P7] (2.6%, 12/463). GII.4 2006b[P4 2006b] predominated in 2011 and 2012. GII.4 Sydney[P31] predominated from 2013 to 2018. In total, 15 genotypes, 15 P-types and 19 dual types were detected in this study, reflecting the genetic diversity. Conclusions There were significant epidemiological characteristics and genetic diversity among outpatient children with norovirus infections < 5 years of age in Beijing from 2011 to 2018. These characteristics differ from those of norovirus outbreaks in Beijing. The complete genome sequences of each genotype are needed to better understand norovirus evolutionary mechanisms.

An adaptive method of defining negative mutation status for multi-sample comparison using next-generation sequencing

Background Multi-sample comparison is commonly used in cancer genomics studies. By using next-generation sequencing (NGS), a mutation's status in a specific sample can be measured by the number of reads supporting mutant or wildtype alleles. When no mutant reads are detected, it could represent either a true negative mutation status or a false negative due to an insufficient number of reads, so-called "coverage". To minimize the chance of false-negative, we should consider the mutation status as "unknown" instead of "negative" when the coverage is inadequately low. There is no established method for determining the coverage threshold between negative and unknown statuses. A common solution is to apply a universal minimum coverage (UMC). However, this method relies on an arbitrarily chosen threshold, and it does not take into account the mutations' relative abundances, which can vary dramatically by the type of mutations. The result could be misclassification between negative and unknown statuses. Methods We propose an adaptive mutation-specific negative (MSN) method to improve the discrimination between negative and unknown mutation statuses. For a specific mutation, a non-positive sample is compared with every known positive sample to test the null hypothesis that they may contain the same frequency of mutant reads. The non-positive sample can only be claimed as “negative” when this null hypothesis is rejected with all known positive samples; otherwise, the status would be “unknown”. Results We first compared the performance of MSN and UMC methods in a simulated dataset containing varying tumor cell fractions. Only the MSN methods appropriately assigned negative statuses for samples with both high- and low-tumor cell fractions. When evaluated on a real dual-platform single-cell sequencing dataset, the MSN method not only provided more accurate assessments of negative statuses but also yielded three times more available data after excluding the “unknown” statuses, compared with the UMC method. Conclusions We developed a new adaptive method for distinguishing unknown from negative statuses in multi-sample comparison NGS data. The method can provide more accurate negative statuses than the conventional UMC method and generate a remarkably higher amount of available data by reducing unnecessary “unknown” calls.

Histamine contents in raw long-ripening meat products commercially available in Poland

Introduction Many consumers seek long-ripening meat products. The availability of these highly distinctive cured pork varieties is continuously expanding and their safety should be subject to monitoring. One of potentially harmful substances in these products is histamine. The presence of this toxic amine is reported in many countries, even in high concentrations. However, the EU has not regulated the permissible histamine content in meat, in a situation at odds with that of regulated fish and fish products. This study established the usefulness of biogenic amine testing in long-ripening pork and furnished indicative concentrations potential useful as a background for future research in preparation for EU regulative intervention. Material and Methods A total of 97 samples of long-ripening meat products untreated by heat were bought from various shops in the Puławy and Lublin regions of Poland and tested for the presence of histamine using high-performance liquid chromatography with diode array. Results The histamine concentration ranged from below limit of detection to 346.64 mg/kg, where 3.47 mg/kg was the lowest in a positive sample. Histamine was detected in 48 samples (49.5%). The maximum amount of histamine was identified in dry ham and the minimum in traditional salami. Conclusion The results of this study suggest that testing meat products for biogenic amines should be a very good indicator of the food safety of long-ripening meats. In half of the tested products, levels of biogenic amines potentially toxic to consumers were determined.

High prevalence of malaria in a non-endemic setting among febrile episodes in travellers and migrants coming from endemic areas: a retrospective analysis of a 2013–2018 cohort

Background The study aimed to analyse the likelihood of imported malaria in people with a suggestive clinical picture and its distinctive characteristics in a hospital in the south of Madrid, Spain. Methods Observational retrospective study that consisted of a review of all medical files of patients with any malaria test registered at Móstoles University Hospital between April 2013 and April 2018. All suspected malaria cases were confirmed by Plasmodium spp. polymerase chain reaction (PCR). Results Of the 328 patients with suspected malaria (53.7% migrant-travellers; 38.7% visitors; 7.6% travellers), 108 cases were confirmed (101 by Plasmodium falciparum), accounting for a 33% positive sample rate. Sixteen cases were diagnosed only by PCR. Patients with malaria, compared to those without, presented predominantly with fever (84% vs. 65%), were older (34 vs. 24 years), sought medical attention earlier (17d vs. 32d), had a greater number of previous malaria episodes (74% vs. 60%), lower levels of platelets (110,500µL vs. 250,000µL), and higher of bilirubin (0.6 mg/dL vs. 0.5 mg/dL). Severe malaria was present in 13 cases; no deaths were recorded. Malaria diagnosis showed a bimodal distribution with two peaks: June to September and November to January. Conclusions Malaria is still a common diagnosis among febrile patients coming from the tropics specially among migrant travellers. Fever, thrombocytopenia, and/or high bilirubin levels should raise suspicion for this parasitic infection. Prompt diagnosis is crucial to avoid severe cases and deaths.

The importance of being heterozygote: effects of RHD-genotype-sex interaction on the physical and mental health of a non-clinical population

Human populations, especially European, are polymorphic in the RHD gene. A significant fraction of their members carry no copy of the coding section of RHD gene, which results in their Rh-negative blood type. Theoretically, this polymorphism should be unstable. Carriers of the less frequent allele are penalized by reduced fertility because of the immunization of RhD-negative mothers by their RhD-positive babies, which results in hemolytic disease of the fetus and newborn in their subsequent progeny. For about 90 years, some form of balancing selection has been suspected to sustain this polymorphism. Several recent studies showed that the RhD-positive heterozygotes express higher viability than both types of homozygotes. However, the genotype of subjects in these studies was estimated only by indirect methods. Here we compared the physical and mental health of 178 women and 86 men who were directly tested for their RHD genotype. The results showed that RhD-positive homozygotic women had worse and RhD-positive homozygotic men better physical health than RhD-negative homozygotes; the difference between RhD-negative homozygotes and heterozygotes was not significant. Our results confirmed that health of RhD-positive heterozygotes and homozygotes differ. Therefore, any result of the comparison of subjects with RhD-positive and RhD-negative phenotype depends on the heterozygote-to-homozygote ratio in the RhD-positive sample. It is, therefore, crucial to analyze the effects of RHD-genotypes, not phenotypes in future studies.

1048. Double-Blind, Randomized, Placebo-Controlled Phase 2b Multicenter Trial of V160, a Replication-Defective Human Cytomegalovirus (CMV) Vaccine

Background Preventing congenital cytomegalovirus infection (CMVi) is an important unmet need. Natural maternal immunity to CMV acquired prior to pregnancy appears to reduce fetal transmission. In a Phase 1 trial, V160, a replication-defective CMV vaccine expressing the pentameric complex, induced humoral and cell-mediated immune (CMI) responses comparable to natural immunity. Methods Healthy, CMV-seronegative women aged 16–35 years were randomized 1:1:1 to receive double-blind V160 in a 3- or 2-dose regimen or placebo. Primary and secondary endpoints were efficacy in reducing the incidence of CMVi with 3-dose or 2-dose regimens of V160 vs placebo, respectively, using a fixed-event design. Monthly urine and saliva samples were collected to identify CMVi by polymerase chain reaction (PCR) with a single positive sample considered evidence of infection. Immunoglobulin G (IgG) binding to glycoprotein B (gB) and CMV-specific neutralizing antibody (NAb) were measured in all participants, and CMI responses were measured in a subset. Injection-site and systemic adverse events (AEs) were collected for 5 days and 14 days, respectively, after each vaccination and serious AEs were collected for the trial duration. Results 2200 women from 7 countries were enrolled (of 7458 screened). Over 80% of participants received all doses, and compliance with saliva and urine samples was &gt; 95%. Vaccine efficacy (VE) of 42.4% (95% CI -13.5, 71.1%) was demonstrated in the 3-dose group vs placebo. In the 2-dose group, VE was -32.0% (95% CI -135.0, 25.0%). Both the quantity and duration of CMV shedding in urine and saliva among cases of CMVi decreased in the 3-dose, but not the 2-dose group vs placebo. Both V160 regimens elicited humoral and CMI responses detected by CMV-specific NAb, gB IgG, and ELISpot, which peaked at Month 7 and continued to be detectable at Month 24. Mild to moderate AEs were more frequently reported in V160 vs placebo recipients, but no vaccine-related serious AEs or deaths were reported. Conclusion V160 was well tolerated and immunogenic, but neither the 3-dose nor 2-dose regimen demonstrated significant efficacy against CMVi as defined in this trial. The quantity and duration of CMV shedding was reduced in the 3-dose group, suggesting V160 may improve immune control of viral replication after CMVi. Disclosures Rituparna Das, MD, Merck & Co, Inc. (Employee) Daniel Blazquez-Gamero, MD, MSD (Other Financial or Material Support, Fees for lectures in educational activities) Soren Gantt, MD, Altona Diagnostics (Research Grant or Support)Merck (Consultant, Grant/Research Support)Meridian Biosciences (Research Grant or Support)Moderna (Consultant, Research Grant or Support)VBI Vaccines Inc (Research Grant or Support) Oliver Bautista, PhD, Merck & Co, Inc. (Employee) Karen Beck, RN, BSN, Merck & Co, Inc. (Employee) Anthony Conlon, PhD, Merck & Co, Inc. (Employee) Daniel Rosenbloom, PhD, Merck & Co, Inc. (Employee) Dai Wang, PhD, Merck & Co, Inc. (Employee) Michael Ritter, BA, Merck & Co, Inc. (Employee) Beth Arnold, MS, Merck & Co, Inc. (Employee, Shareholder) Paula Annunziato, MD, Merck & Co, Inc. (Employee) Kevin Russell, MD, MTM&H, Merck & Co., Inc. (Employee, Shareholder)

187. Characterizing Household Clustering of COVID-19 Cases in Fulton County, Georgia, June 2020–April 2021

Background Households are important for SARS-CoV-2 transmission due to close proximity in enclosed living spaces over long durations. Using contact tracing, the secondary attack rate in households is estimated at 18-20%, yet no studies have examined COVID-19 clustering within households, an important measure to inform testing and prevention. We sought to quantify and characterize household clustering of COVID-19 cases in Fulton County, Georgia. Methods We used state surveillance data to identify all PCR- or antigen-confirmed cases of COVID-19 in Fulton County. Clustered cases were defined as cases with matching street address, including unit number. Communal places (e.g., nursing homes, correctional facilities) were excluded, as were apartments missing unit number. Household clusters were defined as ≥2 COVID-19 cases at the same residential address with positive sample collection dates within 14 days of one another. We described proportion of COVID-19 cases that were clustered, stratified by age, sex, and race/ethnicity over time. Results There were 60,614 COVID-19 cases with available address reported in Fulton County during 6/1/20–4/30/21. Of these, 25,149 (41.6%) had an address that matched at least one other case; 20,793 (34.3%) were from 8,582 household clusters with positive sample collection dates within 14 days (Fig 1). Majority of clusters had 2 individuals (N=6119, 71%), though some had ≥6 individuals (N=79, 0.9%). Clustering increased through January 2021 (Fig 2). Children were more likely to be in household clusters (Fig 4) and 15% of clusters had a child as first diagnosed case with increases since January 2021 (Fig 3). Consistently higher clustering was observed among Hispanic persons, with rising clustering among Asian persons (Fig 5). Figure 1. Distribution of household-clustered COVID-19 cases in Fulton county between June, 2020 and April 2021 Figure 2. Proportion of COVID-19 cases that were part of a household cluster, Fulton County, June 2020–April 2021. Error bars denote 95% confidence interval around the point estimate. Figure 3. Age of first diagnosis among households with at least 2 cases diagnosed within 14 days Conclusion One-third of COVID-19 cases in Fulton County were part of a household cluster. The higher proportion of children in household clusters likely reflects higher probability of living in a home with an adult caregiver. Higher household clustering among Hispanic and Asian persons, regardless of age, may reflect larger households (supported by census data) or increased exposures outside the house. Timely testing for household members to prevent ongoing transmission remains essential. Figure 4. Proportion of COVID-19 cases that were part of a household cluster, by age – Fulton County, June 2020–March 2021 Proportion of COVID-19 cases that were part of a household cluster in Fulton County stratified by race/ethnicity over time Disclosures All Authors: No reported disclosures