Meal kit delivery services in the UK: An evaluation of the nutritional composition of meals.

OBJECTIVE: To examine meals provided by meal kit delivery services (MKDS) and to evaluate their nutritional composition. RESEARCH METHODS AND PROCEDURES: In this cross-sectional study, the nutritional composition of meals (n 497) from MKDS in the UK, was considered. Energy and nutrient content were compared to dietary guidelines; meals were profiled for fat, saturated fat, total sugars, and salt content. RESULTS: There was a large range in the energy and nutrient content of meals. The levels of saturated fat per serving ranged from 0.4 to 28.0 g (Mdn = 9.0 g), and salt content ranged from 0.2 to 6.4 g (Mdn = 2.2 g). Over half of the meals were profiled as high for fat (51.3%), saturated fat (62.2%) and salt (64.4%). Notably, protein content per portion was high (Mdn = 34.0 g), and dietary fiber content was low (Mdn = 6.4 g). Meals, which had been distinguished by the providers with "health-based" descriptors or tags, had a better nutritional profile for fat, saturated fat, and salt, than other meals; nevertheless, many "health-based" meals profiled high for salt (46.5%) and saturated fat (40.4%). CONCLUSIONS: Recipes from MKDS should be revised to improve their nutritional composition; specifically, reductions in salt and saturated fat content and an increase in dietary fiber are needed. Given the variation in the nutritional composition of meals, work is also needed to ascertain the main factors influencing selections made by consumers, and the relevance of guidance and information to support this.

Home Delivery Meal Kits Online Food Safety-Related Information: A Perspective.

The recent popularity of home delivery meal kits (HMK) has prompted concerns about its integrity and safety. On the basis of a food safety-related information evaluation of the common US-based HMK vendors' websites, this perspective highlights opportunities for improvement with the adequacy and accessibility of relevant information on HMK websites, an important resource for communicating food safety best practices to consumers. Identified gaps in information and inadequate delivery protocols potentially increase the risk of offering unsafe food to consumers. Suggestions for future research and recommendations for vendors, policymakers, and regulators to help protect consumers from potential foodborne illness risks are also discussed.

Field validation as a means for continual monitoring of approved test kit's fitness for purpose in the commercial market.

Testing accuracy of a chemical contaminant requires use of a testing platform that conforms to validation criteria outlined in quality literature and standards. This study explores the application of commercial field data measured by qualified analysts using a United States Department of Agriculture - Federal Grain Inspection Service approved kit for measuring fumonisin in maize to augment method validation procedures. Analysts from seven grain testing facilities were qualified in official USDA sampling, sample preparation, and testing methodology using the Charm LF-FUMQ-WETS5. A duplicate sample was tested in the Office of the Texas State Chemist (OTSC) laboratory using UPLC-MS-MS. Data were subject to four statistical techniques using continuous and categorical methodology. This approach enabled researchers to explore if a single test or multiple comparisons were best suited to assess a field kit's fitness for purpose across facility, toxin level, and year. The study concluded that a paired t-test and correlation analysis provided a quick and meaningful evaluation of kit performance. The correct placement of samples within the correct bin (violative versus non-violative) aligns well with market forces and regulatory compliance. The results of this study also provide a useful tool to assess all field kits' performance at the beginning of the harvest season and subsequent years. The combination of statistical techniques presented in this research is an important tool in assessing mycotoxin field test kits fitness for purpose and represents a key step in a continuous improvement-quality systems process meant to protect the feed and food supply.

Safe delivery kits and newborn infection in rural Ethiopian communities.

Objectives: Our goal in this study to investigate the impacts of using safe delivery kits, along with education on their appropriate use, has on preventing newborn and maternal infection. Design: A cross-sectional study. Setting: Participants, and Interventions: we conducted the study on 23 sites across a rural district in Oromia Region, Ethiopia. Safe delivery kits were distributed by health extension workers. Participants comprised 534 mothers between the ages of 17 and 45 years, who were given a safe delivery kit at 7 months' pregnancy for use during their subsequent delivery. Data collection was performed by the trained interviewers in rural Ethiopian communities. Results: Multiple logistic regression analyses showed an independent association between using the cord tie provided in the kits and decreased newborn infection. Specifically, newborns whose mothers used the cord tie were 30 times less likely to develop cord infection than those not using the cord tie in the kits. Further, mothers who received education regarding safe delivery kit use had lower rates of puerperal infection. Conclusion: Single-use delivery kits, when combined with education regarding the appropriate means of using the kit, can decrease the likelihood of maternal infection. Implications for nursing: Nurses and health extension workers in low and middle-income countries should educate mothers on safe delivery kits by providing information regarding their usefulness and the importance of correct and consistent use. Implications for Health Policy: our findings emphasize the need for further interventions in vulnerable countries designed to increase the rate of hygienic birthing practices for deliveries outside health-care facilities.

Comparative assessment of three commercial kits and in house optimized PCR assays for GMO screening in food and feed.

Screening strategies for GMO detection in food and feed are a crucial aspect in GMO testing laboratories for streamlining the analytical workflow and reducing turnaround time and costs. These strategies can be more or less complex or even be targeted according to the ingredients in the product, but whatever the choice, a good basic approach is generally based on the search for 35S promoter (P35S), nos-terminator (T-nos) and FMV promoter (P-FMV). In this study, we compare the singleplex real time PCR method for P35S, T-nos and P-FMV detection currently adopted by the Italian National Reference Laboratory for GM food and feed (NRL) with three commercial kits available on the market for giving a greater choice to consider the best approach suitable to the official control laboratories that are different from each other.•The NRL optimized singleplex PCR methods and the three commercial kits fully respect all the validation parameters criteria according to the minimum performance requirements (MPR) of ENGL [1]•Screening strategies for GMO detection in food and feed are a crucial aspect in GMO testing laboratories and being the commercial kits different from each other, the laboratory can choose the methods best suit their needs reducing turnaround time and costs.

Establishment of a Suitable Diagnostic Workflow to Ensure Sensitive Detection of African Swine Fever Virus Genome in Porcine Semen.

The rapid spread of African swine fever virus (ASFV), causing severe and often lethal disease in domestic pigs and Eurasian wild boar, continues to be a threat to pig populations and dependent industries. Despite scientific achievements that have deepened our understanding of ASFV pathogenesis, alternative transmission routes for ASFV remain to be elucidated. We previously demonstrated the efficient transmission of ASFV from infected boars to naïve recipient gilts via artificial insemination, thereby highlighting the importance of surveillance of boar semen prior to its shipment. Since the accurate and reliable detection of even low amounts of ASFV in boar semen is key to disease prevention and control, we established a suitable diagnostic workflow to efficiently detect the ASFV genome in boar semen. Here, we assessed the sensitivity of various routine nucleic acid extraction kits as well as qPCR protocols in detecting the ASFV genome in the blood and semen of infected boars. The feasibility of the respective kits and methods for future use in boar studs was also considered. Variability in sensitivity mostly concerned samples with low to very low amounts of the ASFV genome. Ultimately, we defined a well-suited workflow for precisely detecting the ASFV genome in boar semen as early as 2 days post ASFV infection.

The role of social media in public health awareness during times of war in Sudan: snakebites and scorpion stings.

BACKGROUND: Snakebite envenomation (SBE) and scorpion sting envenomation (SSE) are significant neglected tropical diseases that primarily affect impoverished communities in rural areas of developing nations. A lack of understanding about snake and scorpion species and their distribution exacerbates the disabilities and fatalities caused by SBE and SSE. In Sudan, particularly in regions affected by ongoing conflicts where healthcare resources are scarce, social media platforms offer a cost-effective approach to addressing public health challenges. Our aim in this study is to highlight the benefits of using social media for data collection and health promotion in such environments. METHODS: We present a cost-effective communication and data collection strategy implemented at the Toxic Organisms Research Centre (TORC) of the University of Khartoum, focusing on a Facebook group, "Scorpions and Snakes of Sudan", as our primary social media platform. Additionally, we discuss the lessons learned and the initial impact of this strategy on enhancing population health literacy. RESULTS: The group community is composed of ~ 5000 members from 14 countries. During the period from January 2023 to January 2024, we received 417 enquiries about snakes and scorpions belonging to 11 families and composed of 55 species. In addition, 53 other enquiries covered a range of organisms and their tracks (e.g., spiders, skinks, chameleons, foxes, sun spiders, centipedes, lizards, moth larvae, and insect tracks). The first photographic evidence of Malpolon monspessulanus in Sudan was via the group activities. The rare species Telescopus gezirae, the Blue Nile cat snake, is also documented via the group member's queries. Recognizing the evolving nature of social media use in public health, we also address the current limitations and evidence gaps that need to be addressed to effectively translate best practices into policy. CONCLUSION: In conclusion, utilizing Facebook as an institutional platform to share scientific information in simple Arabic language underscores the proactive roles that citizens, scientists, and public health stakeholders can play in leveraging social media for eHealth, eAwareness, and public health initiatives. This approach highlights the potential for collaborative efforts, particularly during crises, to maximize the benefits of social media in advancing public health.

Investigating the Impact That Diagnostic Screening with Lateral Flow Devices Had on the Rabies Surveillance Program in Zanzibar, Tanzania.

With the global impetus for the elimination of canine-mediated human rabies, the need for robust rabies surveillance systems has become ever more important. Many countries are working to improve their rabies surveillance programs and, as a result, the reported use of lateral flow devices (LFDs) is increasing. Despite their known diagnostic limitations, previous studies have hypothesised that the benefits associated with LFDs could make them potentially quite useful towards improving the overall robustness of surveillance programs. To test this, a best practice standard operating procedure was developed which was used to guide the implementation of the ADTEC LFD as a diagnostic screening tool in Zanzibar. Over the course of the first 22 months of this investigation, 83 samples were subjected to in-field diagnostic screening, coupled with subsequent laboratory confirmation, and only one false-negative result was detected. Furthermore, the findings of our investigation indicated that the routine use of LFDs as a diagnostic screening tool resulted in a four-fold increase in the number of samples subjected to rabies diagnosis per month and a three-fold increase in the number of wards where samples were collected per year. Our findings suggest that LFDs could play a noteworthy role in improving the robustness of surveillance systems by increasing the number of samples tested and promoting diagnostic screening in areas distant from laboratories. Their implementation would, however, need to be carefully controlled through standardised protocols that align with the international best practices to ensure their judicious use.

An Evaluation of the Sensitivity and Specificity of Three COVID-19 Rapid Immunochromatographic Test Kits Compared to Real-Time Reverse Transcriptase-Polymerase Chain Reaction (rRT-PCR) Among Clinical Samples.

Background and objective The coronavirus disease 2019 (COVID-19) pandemic has imposed a significant burden on healthcare systems worldwide. This highlights the need for simple, rapid, and affordable diagnostic tests that can serve as alternatives to the existing costly and demanding polymerase chain reaction (PCR) assay, especially in resource-limited countries like Ghana. In light of this, we aimed to assess the diagnostic efficacy of three COVID-19 rapid immunochromatographic antigen test kits vs. real-time reverse transcriptase-PCR (rRT-PCR). Methods This study evaluated the sensitivity and specificity of three COVID-19 rapid immunochromatographic antigen test kits: DG Rapid, SD Rapid, and SS Rapid. They were compared with the gold standard RT-PCR for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid antigen in 75 randomly selected archived nasopharyngeal samples. Results Of the 75 samples tested, 38 (50.7%) were positive and 37 (49.3%) were negative for SARS-CoV-2 RNA by rRT-PCR assay. No false positives were recorded. On the other hand, the DG Rapid kit detected 30 (78.9%) true positives and eight (21.1%) false negatives. SD Rapid kit detected 28 (73.7%) true positives and 10 (26.3%) false negatives, while the SS Rapid kit detected 19 (50.0%) true positives and 19 (50.0%) false negatives. While the specificity of each test kit was 100% (95% CI), the sensitivity of the DG Rapid, SD Rapid, and SS Rapid kits was 79%, 74%, and 50% (95% CI), respectively. Higher sensitivities were recorded among samples with cycle threshold (Ct) values <29.99 for each kit. Also, the DG Rapid kit demonstrated 79% excellent agreement with rRT-PCR, while the SD Rapid and SS Rapid kits demonstrated good agreement with rRT-PCR with 73% and 50% Cohen's kappa values, respectively. Conclusions Based on our findings, DG Rapid and SD Rapid kits are reliable alternatives to rRT-PCR for the detection of SARS-CoV-2 infection, especially in resource-limited settings like Ghana.

Enhancing maize safety: the role of co-regulation as a regulatory strategy to manage fumonisin risk.

This study explores the implementation of the One Sample Strategy (OSS), a co-regulation program aimed at managing mycotoxin risk in Texas maize. Fumonisin-contaminated cereals and oilseeds that contain greater than 5 mg kg-1 of the toxin (B1, B2, and B3) are a risk for equids and rabbits, and levels greater than 60 mg kg-1 are a risk to ruminants. The OSS, previously successful in managing aflatoxin risk in Texas maize, was evaluated for its effectiveness in handling fumonisin risk in maize, specifically as it relates to ruminants. In 2017, 25 analysts across seven firms qualified to participate in the program. To ensure greater accuracy in testing, working control samples were provided to the participating OSS firms with the requirement that their results fall within +/- 20% of the target concentration. Ninety-four percent of the working controls met this specification. The capability to grind maize to the OSS prescribed particle size was met by 100% of participants. To verify testing accuracy, file samples collected from each OSS firm were analysed by UPLC-MS/MS. The 177 fumonisin verification samples analysed by Office of the Texas State Chemist (OTSC) were correlated (r = 0.93) with co-regulation laboratories. Results were plotted in an operating curve to depict type I and type II errors. Error analysis revealed a type I error rate of 13% and type II error rate of 2% for the 5 mg kg-1 guidance level, and 6% and 8%, respectively, for the 60 mg kg-1 guidance level. For 2017, 994 official reports of analysis for fumonisin in whole maize in the Texas High Plains were issued by the seven laboratories that employed 25 OTSC-credentialed analysts. The OSS co-regulation program, supported by a quality systems approach and government regulations, has proven effective in managing fumonisin risk in Texas maize, enhancing both market confidence and livestock safety.

Quantitation of milk proteins in thermally treated milk samples and commercial food products by ELISA test kits.

This study evaluated four ELISA kits for quantitation of milk proteins in thermally treated milk samples and food products. How reference materials may be used for comparison of kit performance was examined. Protein contents determined by Veratox Total Milk generally reflected those determined by the 660 nm total protein assay. BioKits BLG Kit was less affected by thermal treatment but resulted in overestimation of protein contents in samples that were boiled, autoclaved or dry-heated at ≤149 °C, while ELISA Systems Casein (ES Casein) and Beta-Lactoglobulin (ES BLG) assays underestimated protein levels in these samples. The four kits gave similar results for ice cream. Veratox registered higher concentrations in all products tested but its sensitivity was greatly lowered in retorted products. ES Casein underperformed Veratox for baked and retorted products. BioKits BLG maintained a better sensitivity towards fried, baked and retorted products while ES BLG exhibited reduced sensitivity for these products.

Cage enrichment to minimize aggression in part-time group-housed female breeding rabbits.

In most rabbit farms, breeding does kindle and nurse their kits in single-litter cages throughout their entire reproduction cycle. However, the protective behavior can lead to aggressive displays and injuries when the does are housed in groups. This study aimed to evaluate cage enrichment for reducing the agonistic behavior in part-time group-housed does. A total of eighty does with their 22-day-old kits were allocated to 20 multi-litter cages, with each cage housing four does and their litters for 10 days. Each multi-litter group was subjected to one of four treatments: alfalfa blocks as distraction material (A), wooden panels underneath the platforms (P), both alfalfa and wooden panels (AP), or no extra enrichment (controls, C). This experiment was replicated for three consecutive reproduction cycles. The skin injuries of the does and the kits were scored with a tagged visual analog scale before grouping and at one, three, six, eight, and 10 days after grouping. Computer vision techniques were used to continuously monitor rabbit activity and agonistic behavior (aggression and fleeing/chasing) during the first 24 h after grouping, specifically during light hours. During the first day in the group, 67.2% of the does and 13.4% of the kits acquired new injuries. This increased to 82.0 and 33.2%, respectively after 10 days in the group relative to the onset of grouping. The injury scores of the does increased toward the sixth day after grouping compared to the first (p < 0.001) and were highest on the tenth day for the kits (p < 0.001). On all the observation days, the number of injured does was higher in C compared to A (p = 0.04) and AP treatment (p = 0.005). There were no other treatment effects observed on the doe or kit skin injuries. Rabbit activity was highest after grouping but decreased after the first and second days (p < 0.001). The agonistic interactions between the does involved more fleeing/chasing behavior (62.0%) rather than aggression (38.0%). Although hierarchy fights are likely when unacquainted does are group-housed, the many animals that sustained injuries and the high injury scores confirm that part-time group housing for does is challenging and possibly inevitable. This study has shown that alfalfa, with or without wooden panels, can slightly reduce the number of injured does.

Unravelling the diagnostic methodologies for SARS-CoV-2; the Indispensable need for developing point-of-care testing.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-caused COVID-19 pandemic that continues to be a global menace and since its emergence in the late 2019, SARS-CoV-2 has been vigorously spreading throughout the globe putting the whole world into a multidimensional calamity. The suitable diagnosis strategies are on the front line of the battle against preventing the spread of infections. Since the clinical manifestation of COVID-19 is shared between various diseases, detection of the unique impacts of the pathogen on the host along with the diagnosis of the virus itself should be addressed. Employing the most suitable approaches to specifically, sensitively and effectively recognize the infected cases may be a real game changer in controlling the outbreak and the crisis management. In that matter, point-of-care assays (POC) appears to be the potential option, due to sensitivity, specificity, affordable, and availability. Here we brief the most recent findings about the virus, its variants, and the conventional methods that have been used for its detection, along with the POC strategies that have been applied to the virus diagnosis and the developing technologies which can accelerate the diagnosis procedure yet maintain its efficiency.

Willingness to Receive HIV Self-Testing Kits from Recent Sexual Partners Among Men in Dar Es Salam, Tanzania: Findings from the STEP Project Baseline Survey.

Globally, men are less likely to access HIV services, and addressing HIV service challenges among men is crucial to the global HIV/AIDS response. HIV self-testing (HIVST) has been shown to be a potentially effective strategy in improving HIV testing coverage among men. This study assessed and identified factors influencing willingness to receive HIVST kits from sexual partners among men in Tanzania. Data are from the baseline survey of the Self-Testing Education and Promotion (STEP) project, a five-year study comprising male participants aged 18 or older who self-reported as HIV-negative. Logistic regression models were used to assess factors associated with men's willingness to receive HIVST kits from their sexual partners. There were 505 heterosexual male participants enrolled in the study with an average age of 29 years, of whom 69% reported being willing to receive HIVST kits from their sexual partner. Logistic regression models demonstrated that willingness to receive HIVST kits from sexual partners was significantly associated with number of sexual partners within 12 months (aOR = 1.2, 95% CI [1.1-1.3]), awareness of HIVST (aOR = 5.6, 95% CI [3.2-9.5]), previous discussion of HIVST with sexual partners aOR = 14.0, 95% CI [8.0-24.6]), and previous testing for HIV with sexual partners not (aOR = 2.5, 95% CI [1.3-4.7]). These findings suggest additional promotional strategies to improve men's awareness of HIVST and support open conversations about HIVST and HIV testing with sexual partners could improve men's willingness to receive HIVST kits when distributed through their sexual partners.

Evaluation of the effect of Carica papaya seed on the growth performance of fattening rabbits.

Successful breeding depends on feeding. The present study aims to evaluate the Carica papaya seed effect on the growth performance of rabbits. The zootechnical parameters studied are weight growth, average daily gain, Feed Conversion Ratio, and carcass characteristics of kits. The experiment was conducted on 48 rabbits, divided into 4 groups, for 6 weeks. Forty-eight rabbits were divided into four (04) groups of 3 repetitions of 4 rabbits. The animals were fed diets containing various levels of papaya seed powder at variable contents: 0% (group T0), 4% (group T1), 6% (group T2), and 8% (group T3). At the end of the experiment, three animals were slaughtered in each animal group to assess the quality of the carcasses and organs. 6% of the seeds of Carica papaya significantly improved (p < 0.05) the average daily gain of the kits: T2 (22.40 g / d) compared to the T0 group (11.32 g / d), T1 (12.20 g / d) and T3 (17.53 g / d). The best Feed Conversion Ratio (0.80) was recorded in the animals of group T2. In contrast, the highest carcass yield was recorded in the rabbits of group T3 (62.70%). In conclusion, 6% was optimal in the feed rations of fattened rabbits to improve production performance. Breeders can consider the benefits of introducing Carica papaya seeds into the rabbits' diet.

Comparison of Fecal MicroRNA Isolation Using Various Total RNA Isolation Kits.

Fecal specimens have long been regarded as promising sources for gastrointestinal cancer screening and have, thus, been extensively investigated in biomarker research. MicroRNAs (miRNAs) are small, non-coding RNA molecules involved in regulating various biological processes. They are commonly dysregulated during tumor development and exhibit differential expression in feces. To assess the preanalytical feasibility of fecal miRNA analysis, we systematically compared the performance of commonly used total RNA extraction methods. Fecal samples from healthy subjects were utilized for this evaluation. Various methods, including miRNeasy, Universal, Trizol, RNeasy, and mirVana kits, were employed to isolate total RNA. MiRNA expression analyses were conducted using TaqMan or SYBR Green qRT-PCR for a subset of miRNAs, with externally spiked-in cel-miR-39 used for normalization. Most methods demonstrated similar performance in terms of the total RNA concentration and purity. Externally spiked cel-miR-39 and endogenous miRNAs (RNU6b, miR-16, and miR-21) exhibited comparable concentrations across the different RNA isolation methods, whereas the RNeasy mini kit consistently yielded lower values. Our findings suggest that various isolation methods produce reproducible and comparable miRNA expression results, supporting the potential comparability and translational applicability of miRNA-based biomarker research in the future.

Validation of a new kit for preeclampsia screening: A comprehensive analysis.

Objectives: Preeclampsia is a common pregnancy complication that significantly contributes to maternal mortality, perinatal mortality, and preterm delivery. The sFlt-1/PlGF (fms-like tyrosine kinase-1/placental growth factor) ratio has demonstrated robust diagnostic value for preeclampsia. This study assessed the analytical performance and diagnostic accuracy of a novel quantitative determination kit for sFlt-1 and PlGF for the diagnosis of preeclampsia. Methods: The detection performance of the test kit was validated using the Center for Medical Device Evaluation (CMDE) and Clinical and Laboratory Standards Institute (CLSI) documents. The test results were compared to those of the Elecsys immunoassay (Roche Diagnostics). Independent discovery and validation sets were used to analyze the diagnostic efficacy of the preeclampsia kit. The area under the curve (AUC) for preeclampsia at different gestational ages was calculated. Results: Correlation analysis between the test and Roche kits revealed a strong concordance (sFlt-1: r = 0.9966, P < 0.0001; PlGF: r = 0.9935, P < 0.0001). The AUCs for sFlt-1, PlGF, and the sFlt-1/PlGF ratio in diagnosing preeclampsia were 0.749, 0.795, and 0.834, respectively, in the discovery set and 0.729, 0.811, and 0.831, respectively, in the validation set. The corresponding results from the Roche kit were 0.741, 0.795, and 0.829, respectively, and 0.761, 0.864, and 0.844, respectively. Conclusions: Quantitative sFlt-1 and PlGF kits exhibited high levels of consistency with the Roche kits in terms of quantitative outcomes and diagnostic performance for preeclampsia.

Predicting probative levels of touch DNA on tapelifts using Diamond™ Nucleic Acid Dye.

Tapelifting is a common strategy to recover touch DNA deposits from porous exhibits in forensic DNA casework. However, it is known that only about 30 % of tapelifts submitted for DNA analysis in operational forensic laboratories yield profiles suitable for comparison or upload to a searchable database. A reliable means to identify and remove non-probative tapelifts from the workflow would reduce sample backlogs and provide significant cost savings. We investigated whether the amount of macroscopic or microscopic fluorescence on a tapelift following staining with Diamond Nucleic Acid Dye (DD), determined using a Polilight and Dino Lite microscope respectively, could predict the DNA yield and/or the DNA profiling outcome using controlled (saliva), semi-controlled (finger mark) and uncontrolled (clothing) samples. Both macroscopic and microscopic DD fluorescence could predict DNA yield and profiling outcome for all sample types, however the predictive power deteriorated as the samples became less controlled. For tapelifts of clothing, which are operationally relevant, Polilight fluorescence scores were significantly impacted by clothing fibres and other non-cellular debris and could not be used to identify non-probative samples. The presence of less than 500 cells on a clothing tapelift using microscopic counting of stained corneocytes was identified as a potential threshold for a non-probative DNA profiling outcome. A broader examination of the reliability of this threshold using a casework trial is recommended. Due to the labour intensiveness of microscopic cell counting, and the increased risk of inadvertent contamination, automation of this process using image software in conjunction with artificial neural networks (ANN) should be explored.

Cell-Free Microbial DNA Analysis: Effects of Blood Plasma and Serum Quantity, Biobanking Protocols, and Isolation Kits.

Recent studies highlight the presence of bacterial sequences in the human blood, suggesting potential clinical significance for circulating microbial signatures. These sequences could presumably serve in the diagnosis, prediction, or monitoring of various health conditions. Ensuring the similarity of samples before bacterial analysis is crucial, especially when combining samples from different biobanks prepared under varying conditions (such as different DNA extraction kits, centrifugation conditions, blood collection tubes, etc.). In this study, we aimed to analyze the impact of different sample collection and nucleic acid extraction criteria (blood collection tube, centrifugation, input volume, and DNA extraction kit) on circulating bacterial composition. Blood samples from four healthy individuals were collected into three different sample collection tubes: K2EDTA plasma tube, sodium citrate plasma tube, and gel tube for blood serum. Tubes were centrifugated at standard and double centrifugation conditions. DNA extraction was performed using 100, 200, and 500 µL plasma/serum input volumes. DNA extraction was performed using three different isolation kits: Norgen plasma/serum cell-free circulating DNA purification micro kit, Applied Biosystems MagMAX cell-free DNA isolation kit, and Qiagen QIAamp MinElute cell-free circulating DNA mini kit. All samples were subjected to 16S rRNA V1-V2 library preparation and sequencing. In total, 216 DNA and 18 water control samples were included in the study. According to PERMANOVA, PCoA, Mann-Whitney, and FDR tests the effect of the DNA extraction kit on the microbiota composition was the greatest, whereas the type of blood collection tube, centrifugation type, and sample input volume for the extraction had minor effects. Samples extracted with the Norgen DNA extraction kit were enriched with Gram-negative bacteria, whereas samples extracted with the Qiagen and MagMAX kits were enriched with Gram-positive bacteria. Bacterial profiles of samples prepared with the Qiagen and MagMAX DNA extraction kits were more similar, whereas samples prepared with the Norgen DNA extraction kit were significantly different from other groups.