Pathogen Reduction Research Articles

Effects of pre-freeze pathogen reduction with riboflavin and UV light on red cells stored post-thaw in AS-3 additive solution.

Pathogen reduction technology (PRT) may improve the safety of RBCs for transfusion. As the Czech Republic considers PRT, we asked what effects riboflavin and UV light PRT pre-freezing has on the post-thaw recovery and properties of cryopreserved RBCs (CRBCs) after deglycerolization and liquid storage. 24 Group O whole blood (WB) units were leukoreduced and then treated with riboflavin and UV light PRT (Mirasol, Terumo BCT, USA) before cryopreservation (T-CRBC); 20 similarly-collected units were untreated controls (C-CRBC). Units were processed to RBCs and then cryopreserved with 40% glycerol (wt/vol), frozen at -80°C, stored >118 days, reconstituted as deglycerolized RBC units in AS-3, and stored at 4 ± 2°C for 21 days. One treated unit sustained massive hemolysis during the post-thaw wash process and was removed from data analysis. The remaining units were assessed pre-PRT, post-PRT, and post-thaw-wash on days 0, 7, 14, and 21 for hematocrit, volume, hemoglobin per transfusion unit, pH, % hemolysis, hemoglobin in the supernatant, potassium, phosphorus, NH3 , osmolality, ATP, and 2,3-diphosphoglycerate. PRT with leukoreduction caused a 5% loss of RBC followed by a 24% freeze-thaw-wash related loss for a total 28% loss but treated units contained an average of 45 g of hemoglobin, meeting European Union guidelines for CRBC. T-CRBCs displayed higher post-wash hemolysis, potassium, and ammonia concentrations, and lower ATP at the end of storage. Cryopreserved RBCs from Riboflavin and UV light-treated WB meet the criteria for clinical use for 7 days after thawing and provide additional protection against infectious threats.

Response to intranasal Lactococcus lactis W136 probiotic supplementation in refractory CRS is associated with modulation of non-type 2 inflammation and epithelial regeneration.

We have previously documented that in individuals with chronic rhinosinusitis (CRS) refractory to surgery, intranasal application of live Lactococcus lactis W136, a probiotic bacterium, improves sinus-specific symptoms, SNOT-22, and mucosal aspect on endoscopy, accompanied by a reduction in sinus pathogens and an increase in protective bacteria. The present work explores the molecular mechanisms underpinning these observations using transcriptomics of the sinus mucosa. Epithelial brushings collected prospectively as a sub-study of the L. lactis W136 clinical trial were used to probe epithelial responses to microbiome supplementation using a hypothesis-free bioinformatic analysis of gene expression analysis. Samples from twenty-four patients with CRS refractory to medical and surgical management were prospectively collected during a clinical trial assessing the effect of 14 days of BID nasal irrigation with 1.2 billion CFU of live L. lactis W136 probiotic bacteria (CRSwNP = 17, CRSsNP = 7). Endoscopically guided sinus brushings were collected as part of the initial study, with brushings performed immediately before and after treatment. Following RNA extraction, samples were assessed using the Illumina HumanHT-12 V4 BeadChip. Differential gene expression was calculated, and pathway enrichment analysis was performed to identify potentially implicated processes. Differentially identified transcripts and pathways were assessed for the overall population and the clinical phenotypes of CRSwNP and CRSsNP. Patterns of response to treatment were similar across all groups, implicating pathways for the regulation of immunity and epithelial cell regulation. These resemble the patterns of improvement observed following successful treatment with endoscopic sinus surgery or azithromycin. Gene expression profiling following the application of live bacteria to the diseased sinus epithelium highlights the implication of multiple components of the inflammation-microbiome-epithelial barrier axis implicated in CRS. These effects appear to involve both epithelial restoration and modulation of innate and adaptive immunity, supporting the potential interest of targeting the sinus epithelium and the microbiome as potential CRS therapies.

The Influence of the Multi-Component Mineral-Organic Concentrate on the Bonitation Value of Turfgrass

Multi-component fertilization has been found to have effects on grass metabolism, such as the stimulation of life processes and the reduction of adverse environmental conditions and pathogens. The research aimed to determine the bonitation value (assessment of the value in use) of turfgrass under the influence of using a multi-ingredient fertilizer. The experiment was carried out at the Experimental Station of the University of Agriculture in Krakow (Poland). The solution was applied through foliar application at three rates: 1.0, 2.0, and 3.0 L·ha−1. This fertilizer contains essential minerals and growth stimulants. An increase in the concentration of the test fertilizer used for spraying was associated with increased effectiveness. The plants with the highest dose of the multi-component fertilizer (treatment III) were characterized by the highest aesthetic values. The use of the concentrate reduced the occurrence of fungal plant diseases. Compared to control plants, 13% less snow mold infection and 25% fewer brown leaf spots were found. Satisfactory effects were also obtained on objects where mineral-organic concentrate was applied at a dose of 2.0 L·ha−1 (treatment II). Plants that received Treatment II and III resulted in 9% less snow mold and 15% less brown leaf spot compared to controls. In the object with the highest concentration dose (treatment III), the green index (NDVI) was also higher by 8% and the leaf greenness index (SPAD) by 7% compared to the plants from the control objects.

PATTERNS OF SEROPREVALENCE FOR TRANSFUSION-TRANSMISSIBLE INFECTIONS AMONG BLOOD DONORS IN A BLOOD CENTRE OF NORTHEAST INDIA

Introduction: Transfusion– Transmissible Infections (TTIs) are the infections resulting from the introduction of a pathogen into a person through blood transfusion. In an attempt to mitigate the inherent risk of TTIs, the demographic information of blood donors including knowledge on local prevalence of infections and its pattern in the donor population is important for formulating recruitment strategies and planning other precautionary measures. Materials & Methods: This cross-sectional study was conducted in the Blood Centre, Jawaharlal Nehru Institute of Medical Sciences, Imphal between January 2019 and December 2022. All donated blood units were tested for the mandatory TTI markers for HIV 1 & 2, Hepatitis B, Hepatitis C, Syphilis and Malarial parasite. The prevalence and patterns of seropositivity for TTIs were studied based on the donor demographic characteristics such as donation type, gender and age group. Results: The overall seroprevalence among blood donors (n=31,563) was 1.79%. The seroprevalence was lower among voluntary donors (1.08%) than the replacement donors (2.10%). Seropositivities among female and male donors were 0.79% and 1.93% respectively. The lowest TTI seropositivity was seen among female voluntary donors (0.59%) and highest among male replacement donors (2.24%). The seroprevalence for specic TTI markers among the blood donors were 0.15% for HIV; 0.57% for HBV; 0.93% for HCV; 0.14% for Syphilis and no donor was found positive for Malaria. The frequency of TTI seropositivity was increased with increase in the age groups of donors in both sexes. There were 9 (0.029%) donors who had co-infection of TTIs. The patterns of TTI markers for co-infections were HCV+HIV=3; HCV+HBV=3; HCV+Syphilis=2 and HIV+ Syphilis=1. Conclusion: The measures to provide safe blood may include collection of blood from the targeted low risk donor population, using more sensitive testing methods, implementing pathogen reduction technologies and other public health measures.

Assessment of the impact of pathogen reduction technologies on the neutralizing activity of COVID-19 convalescent plasma.

COVID-19 convalescent plasma (CCP) could improve the clinical outcome of COVID-19 patients when high-titer CCP is administered in early stages of disease. However, CCP donors have a risk profile like first-time donors, pathogen reduction treatment (PRT) may mitigate such risk but should not impact CCP quality. The current study aims to assess the impact of PRT-technologies available in Saudi Arabia on the neutralizing activity of CCP. Study designand Methods: CCP was collected from eligible donors by plasmapheresis. The neutralization titer was determined with an in-house microneutralization assay (MNA) using a local SARS-CoV-2 clinical isolate. Selected units were split and subject to PRT with amotosalen/UVA (AS) or Riboflavin/UVB (RB) (pairwise side-by-side comparison) followed by a second MNA analysis. 51 CCP units were collected, 27 were included in the analysis reaching the minimum MNA titer of 1:40 (4 reached high titer (≥1:250)). 27 CCP units were treated with AS and 14 with RB, the median MNA pre-treatment titer was 1:80 (1:40–640). The impact of AS and RB PRT on CCP neutralizing activity was not significantly different, nor in the total analysis neither in the pairwise comparison (94.6 vs 96.4 % retention, p > 0.05). No correlation of titer and blood group was observed, but a trend for increasing MNA titer with donor age, choosing donors with an age > 45 years would increase the number of high-titer CCP donors. The difference in impact of AS and RB on CCP MNA titer was below the limit of detection of the assay (0.5-fold).

Metagenomic analysis examines oral microbiome changes and interplay with immune response following prenatal total oral rehabilitation

BackgroundSuboptimal maternal oral health during pregnancy is potentially associated with adverse birth outcomes and increased dental caries risks in children. This study aimed to assess the oral microbiome and immune response following an innovative clinical regimen, Prenatal Total Oral Rehabilitation (PTOR), that fully restores women’s oral health to a “disease-free status” before delivery.MethodsThis prospective cohort study assessed 15 pregnant women at baseline and 3 follow-up visits (1 week, 2 weeks, and 2 months) after receiving PTOR. The salivary and supragingival plaque microbiomes were analyzed using metagenomic sequencing. Multiplexed Luminex cytokine assays were performed to examine immune response following PTOR. The association between salivary immune markers and oral microbiome was further examined.ResultsPTOR was associated with a reduction of periodontal pathogens in plaque, for instance, a lower relative abundance of Tannerella forsythia and Treponema denticola at 2 weeks compared to the baseline (p < 0.05). The alpha diversity of plaque microbial community was significantly reduced at the 1-week follow-up (p < 0.05). Furthermore, we observed significant changes in the Actinomyces defective-associated carbohydrate degradation pathway and Streptococcus Gordonii-associated fatty acid biosynthesis pathway. Two immune markers related to adverse birth outcomes significantly differed between baseline and follow-up. ITAC, negatively correlated with preeclampsia severity, significantly increased at 1-week follow-up; MCP-1, positively correlated with gestational age, was elevated at 1-week follow-up. Association modeling between immune markers and microbiome further revealed specific oral microorganisms that are potentially correlated with the host immune response.ConclusionsPTOR is associated with alteration of the oral microbiome and immune response among a cohort of underserved US pregnant women. Future randomized clinical trials are warranted to comprehensively assess the impact of PTOR on maternal oral flora, birth outcomes, and their offspring’s oral health.

Advances in pulsed electric stimuli as a physical method for treating liquid foods.

There is a need for alternative technologies that can deliver safe and nutritious foods at lower costs as compared to conventional processes. Pulsed electric field (PEF) technology has been utilised for a plethora of different applications in the life and physical sciences, such as gene/drug delivery in medicine and extraction of bioactive compounds in food science and technology. PEF technology for treating liquid foods involves engineering principles to develop the equipment, and quantitative biochemistry and microbiology techniques to validate the process. There are numerous challenges to address for its application in liquid foods such as the 5-log pathogen reduction target in food safety, maintaining the food quality, and scale up of this physical approach for industrial integration. Here, we present the engineering principles associated with pulsed electric fields, related inactivation models of microorganisms, electroporation and electropermeabilization theory, to increase the quality and safety of liquid foods; including water, milk, beer, wine, fruit juices, cider, and liquid eggs. Ultimately, we discuss the outlook of the field and emphasise research gaps.

Risk of Gastroenteritis from Swimming at a Wastewater-Impacted Tropical Beach Varies across Localized Scales.

Population growth and changing climate are expected to increase human exposure to pathogens in tropical coastal waters. We examined microbiological water quality in three rivers within 2.3 km of each other that impact a Costa Rican beach and in the ocean outside their plumes during the rainy and dry seasons. We performed quantitative microbial risk assessment (QMRA) to predict the risk of gastroenteritis associated with swimming and the amount of pathogen reduction needed to achieve safe conditions. Recreational water quality criteria based on enterococci were exceeded in >90% of river samples but in only 13% of ocean samples. Multivariate analysis grouped microbial observations by subwatershed and season in river samples but only by subwatershed in the ocean. The modeled median risk from all pathogens in river samples was between 0.345 and 0.577, 10-fold above the U.S. Environmental Protection Agency (U.S. EPA) benchmark of 0.036 (36 illnesses/1,000 swimmers). Norovirus genogroup I (NoVGI) contributed most to risk, but adenoviruses raised risk above the threshold in the two most urban subwatersheds. The risk was greater in the dry compared to the rainy season, due largely to the greater frequency of NoVGI detection (100% versus 41%). Viral log10 reduction needed to ensure safe swimming conditions varied by subwatershed and season and was greatest in the dry season (3.8 to 4.1 dry; 2.7 to 3.2 rainy). QMRA that accounts for seasonal and local variability of water quality contributes to understanding the complex influences of hydrology, land use, and environment on human health risk in tropical coastal areas and can contribute to improved beach management. IMPORTANCE This holistic investigation of sanitary water quality at a Costa Rican beach assessed microbial source tracking (MST) marker genes, pathogens, and indicators of sewage. Such studies are still rare in tropical climates. Quantitative microbial risk assessment (QMRA) found that rivers impacting the beach consistently exceeded the U.S. EPA risk threshold for gastroenteritis of 36/1,000 swimmers. The study improves upon many QMRA studies by measuring specific pathogens, rather than relying on surrogates (indicator organisms or MST markers) or estimating pathogen concentrations from the literature. By analyzing microbial levels and estimating the risk of gastrointestinal illness in each river, we were able to discern differences in pathogen levels and human health risks even though all rivers were highly polluted by wastewater and were located less than 2.5 km from one another. This variability on a localized scale has not, to our knowledge, previously been demonstrated.

Efficacy of Antimicrobial Spray Treatments in Reducing Salmonella enterica Populations on Chilled Pork

Studies reporting on alternative antimicrobial interventions for pathogen control on chilled pork carcasses and cuts are limited. In this study, the antimicrobial effects of various spray treatments against Salmonella enterica inoculated on skin-on pork samples were evaluated. Chilled pork jowls were portioned (10 by 5 by 1 cm) and inoculated, on the skin side, with a mixture of six S. enterica serotype strains to target levels of 6 to 7 log CFU/cm2 (high inoculation level) or 3 to 4 log CFU/cm2 (low inoculation level). Samples were then left nontreated (control) or were treated (10 s) using a laboratory-scale spray cabinet with water, formic acid (1.5%), a proprietary blend of sulfuric acid and sodium sulfate (SSS, pH 1.2), peroxyacetic acid (PAA, 400 ppm), or PAA (400 ppm) that was pH-adjusted (acidified) with acetic acid (1.5%), formic acid (1.5%), or SSS (pH 1.2). Samples (n = 6) were analyzed for Salmonella populations after treatment application (0 h) and after 24 h of refrigerated (4°C) storage. Irrespective of inoculation level, all spray treatments effectively reduced (P < 0.05) Salmonella levels immediately following their application. Overall, pathogen reductions for the chemical treatments, compared to the respective high and low inoculation level nontreated controls, ranged from 1.2 to 1.9 log CFU/cm2 (high inoculation level) and 1.0 to 1.7 log CFU/cm2 (low inoculation level). Acidification of PAA with acetic acid, formic acid, or SSS did not (P ≥ 0.05) enhance the initial bactericidal effects of the nonacidified PAA treatment. Salmonella populations recovered from all treated samples following 24 h of storage were, in general, similar (P ≥ 0.05) or up to 0.6 log CFU/cm2 lower (P < 0.05) than those recovered from samples analyzed immediately after treatment application. The results of the study may be used by processing establishments to help identify effective decontamination interventions for reducing Salmonella contamination on pork.

Efficacy of Common Antimicrobial Interventions at and above Regulatory Allowable Pick-Up Levels on Pathogen Reduction.

The objective of this study was to evaluate the food safety efficacy of common antimicrobial interventions at and above required uptake levels for processing aids on the reduction of Shiga-toxin producing E. coli (STEC) and Salmonella spp. through spray and dip applications. Beef trim was inoculated with specific isolates of STEC or Salmonella strains. Trim was intervened with peracetic or lactic acid through spray or dip application. Meat rinses were serially diluted and plated following the drop dilution method; an enumerable range of 2-30 colonies was used to report results before log transformation. The combination of all treatments exhibits an average reduction rate of 0.16 LogCFU/g for STEC and Salmonella spp., suggesting that for every 1% increase in uptake there is an increase of 0.16 LogCFU/g of reduction rate. There is a statistical significance in the reduction rate of Shiga-toxin producing Escherichia coli in relation to the uptake percentage (p < 0.01). The addition of explanatory variables increases the R2 of the regression for STEC, where all the additional explanatory variables are statistically significant for reduction (p < 0.01). The addition of explanatory variables increases the R2 of the regression for Salmonella spp., but only trim type is statistically significant for reduction rate (p < 0.01). An increase in uptake percentages showed a significant increase in reduction rate of pathogens on beef trimmings.

The Evolution of the Safety of Plasma Products from Pathogen Transmission-A Continuing Narrative.

Chronic recipients of plasma products are at risk of infection from blood-borne pathogens as a result of their inevitable exposure to agents which will contaminate a plasma manufacturing pool made up of thousands of individual donations. The generation of such a pool is an essential part of the large-scale manufacture of these products and is required for good manufacturing practice (GMP). Early observations of the transmission of hepatitis by pooled plasma and serum led to the incorporation of heat treatment of the albumin solution produced by industrial Cohn fractionation of plasma. This led to an absence of pathogen transmission by albumin over decades, during which hepatitis continued to be transmitted by other early plasma fractions, as well as through mainstream blood transfusions. This risk was decreased greatly over the 1960s as an understanding of the epidemiology and viral aetiology of transfusion-transmitted hepatitis led to the exclusion of high-risk groups from the donor population and the development of a blood screening test for hepatitis B. Despite these measures, the first plasma concentrates to treat haemophilia transmitted hepatitis B and other, poorly understood, forms of parenterally transmitted hepatitis. These risks were considered to be acceptable given the life-saving nature of the haemophilia treatment products. The emergence of the human immunodeficiency virus (HIV) as a transfusion-transmitted infection in the early 1980s shifted the focus of attention to this virus, which proved to be vulnerable to a number of inactivation methods introduced during manufacture. Further developments in the field obviated the risk of hepatitis C virus (HCV) which had also infected chronic recipients of plasma products, including haemophilia patients and immunodeficient patients receiving immunoglobulin. The convergence of appropriate donor selection driven by knowledge of viral epidemiology, the development of blood screening now based on molecular diagnostics, and the incorporation of viral inactivation techniques in the manufacturing process are now recognised as constituting a "safety tripod" of measures contributing to safety from pathogen transmission. Of these three components, viral inactivation during manufacture is the major contributor and has proven to be the bulwark securing the safety of plasma derivatives over the past thirty years. Concurrently, the safety of banked blood and components continues to depend on donor selection and screening, in the absence of universally adopted pathogen reduction technology. This has resulted in an inversion in the relative safety of the products of blood banking compared to plasma products. Overall, the experience gained in the past decades has resulted in an absence of pathogen transmission from the current generation of plasma derivatives, but maintaining vigilance, and the surveillance of the emergence of infectious agents, is vital to ensure the continued efficacy of the measures in place and the development of further interventions aimed at obviating safety threats.

Peroxin Pex14/17 Is Required for Trap Formation, and Plays Pleiotropic Roles in Mycelial Development, Stress Response, and Secondary Metabolism in Arthrobotrys oligospora.

The peroxins encoded by PEX genes involved in peroxisome biogenesis play a crucial role in cellular metabolism and pathogenicity in fungi. Herein, we characterized a filamentous fungus-specific peroxin Pex14/17 in the Arthrobotrys oligospora, a representative species of nematode-trapping fungi. The deletion of AoPEX14/17 resulted in a remarkable reduction in mycelial growth, conidia yield, trap formation, and pathogenicity. Compared with the wild-type strain, the ΔAopex14/17 mutant exhibited more lipid droplet and reactive oxygen species accumulation accompanied with a significant decrease in fatty acid utilization and tolerance to oxidative stress. Transcriptomic analysis indicated that AoPEX14/17 was involved in the regulation of metabolism, genetic information processing, environmental information processing, and cellular processes. In subcellular morphology, the deletion of AoPEX14/17 resulted in a decrease in the number of cell nuclei, autophagosomes, and Woronin bodies. Metabolic profile analysis showed that AoPex14/17 affects the biosynthesis of secondary metabolites. Yeast two-hybrid assay revealed that AoPex14/17 interacted with AoPex14 but not with AoPex13. Taken together, our results suggest that Pex14/17 is the main factor for modulating growth, development, and pathogenicity in A. oligospora. IMPORTANCE Peroxisome biogenesis genes (PEX) play an important role in growth, development, and pathogenicity in pathogenic fungi. However, the roles of PEX genes remain largely unknown in nematode-trapping (NT) fungi. Here, we provide direct evidence that AoPex14/17 regulates mycelial growth, conidiation, trap formation, autophagy, endocytosis, catalase activity, stress response to oxidants, lipid metabolism, and reactive oxygen species production. Transcriptome analysis and metabolic profile suggested that AoPex14/17 is involved in multiple cellular processes and the regulation of secondary metabolism. Therefore, our study extends the functions of PEX genes, which helps to elucidate the mechanism of organelle development and trap formation in NT fungi and lays the foundation for the development of efficient nematode biocontrol agents.

Effect of germicidal short wavelength ultraviolet light on the polyphenols, vitamins, and microbial inactivation in a highly opaque apple juice

AbstractThe aim of this investigation was to study the efficacy of UV‐C light emitting diode (LED) system operating at 263 nm for the inactivation of Listeria monocytogenes and Escherichia coli O157:H7. UV‐C irradiation was applied to stirred 5 ml samples, using a collimated beam LED system operating at a 263 nm wavelength. UV irradiation doses ranging from 0 to 160 mJ/cm2 were also delivered to apple juice (AJ) and polyphenols and vitamins were profiled. Liquid chromatography–mass spectrometry (LC–MS/MS) analysis was conducted to assess the stability of polyphenols or vitamins in UV‐C exposed AJ. The polyphenol and vitamin results demonstrated that UV‐C irradiation in AJs at relevant commercial UV doses induced significant reductions in the concentrations of selected polyphenols and vitamins, p &lt; .05. Ascorbic acid was reduced to 32%, at 160 mJ/cm2 whereas 17% reduction was observed at 40 mJ/cm2. Riboflavin was observed to be relatively stable. Epicatechin and chlorogenic were significantly reduced at high exposure doses. In contrast, minor changes were observed at 40 mJ/cm2. Results show that UV‐C irradiation effectively inactivated pathogenic microbes in AJ. Inactivation levels of all organisms were proportional to UV‐C dose. The log reduction kinetics of microorganisms followed log‐linear and with higher R2 (&gt;0.95). The decimal reduction dose (D10) values of 4.16 and 3.84 mJ/cm2 were obtained from the inactivation of E. coli and L. monocytogenes in AJ. The results from this study imply that adequate log reduction of pathogens is achievable in AJ and suggests significant potential for UV‐C treatment of other opaque liquid foods.

Underestimation of Per- and Polyfluoroalkyl Substances in Biosolids: Precursor Transformation During Conventional Treatment.

Wastewater treatment plants generate a solid waste known as biosolids. The most common management option for biosolids is to beneficially reuse them as an agricultural amendment, but because of the risk of pathogen exposure, many regulatory bodies require pathogen reduction before biosolids reuse. Per- and polyfluoroalkyl substances (PFAS) are well documented in biosolids, but limited information is available on how biosolids treatment processes impact PFAS. Furthermore, quantification of PFAS has focused on perfluoroalkyl acids (PFAAs) which are a small fraction of thousands of PFAS known to exist. The objective of this study was to quantify 92 PFAS in biosolids collected from eight biosolids treatment facilities before and after four pathogen treatment applications: composting, heat treatment, lime treatment, and anaerobic digestion. Overall, total PFAS concentrations before and after treatment were dominated by PFAA precursor species, in particular, diPAPs which accounted for a majority of the mass of the Σ92PFAS. This differs from historic data that found PFAAs, primarily PFOS, to dominate total PFAS concentrations. Treatment options such as heat treatment and composting changed the ratio of PFAA precursors to PFAAs indicating a transformation of PFAS during treatment. This study finds that PFAA precursors are likely underrepresented by other studies and make up a larger percentage of the total PFAS concentration in biosolids than previously estimated.

Pathogen reduction with methylene blue does not have an impact on the clinical effectiveness of COVID-19 convalescent plasma.

There is a concern about a possible deleterious effect of pathogen reduction (PR) with methylene blue (MB) on the function of immunoglobulins of COVID-19 convalescent plasma (CCP). We have evaluated whether MB-treated CCP is associated with a poorer clinical response compared to other inactivation systems at the ConPlas-19 clinical trial. This was an ad hoc sub-study of the ConPlas-19 clinical trial comparing the proportion of patients transfused with MB-treated CCP who had a worsening of respiration versus those treated with amotosalen (AM) or riboflavin (RB). One-hundred and seventy-five inpatients with SARS-CoV-2 pneumonia were transfused with a single CCP unit. The inactivation system of the CCP units transfused was MB in 90 patients (51.4%), RB in 60 (34.3%) and AM in 25 (14.3%). Five out of 90 patients (5.6%) transfused with MB-treated CCP had worsening respiration compared to 9 out of 85 patients (10.6%) treated with alternative PR methods (p=0.220). Of note, MB showed a trend towards a lower rate of respiratory progressions at 28 days (risk ratio, 0.52; 95% confidence interval, 0.18-1.50). Our data suggest that MB-treated CCP does not provide a worse clinical outcome compared to the other PR methods for the treatment of COVID-19.

How do we forecast tomorrow's transfusion: Infectious safety?

Continuous improvement has led to a very high degree of microbial safety of transfusion. Four issues are likely to impact the future of this safety. There will be further advances in the efficacy and efficiency of donation testing and pathogen reduction, increasing safety and hopefully eliminating unnecessary procedures. While system failures have been essentially eliminated, there will be ongoing, unpredictable issues that are inevitable. Emerging infections are likely the greatest concern and will continue, although advances in science and technology will permit increasingly rapid responses to outbreaks. Finally, the practice of transfusion may eventually impact safety as usage of blood is reduced and perhaps as alternatives to conventional blood components are developed.

Pathogen reduction of monkeypox virus in plasma and whole blood using riboflavin and UV light.

Monkeypox virus has recently emerged from endemic foci in Africa and, since October 20, 2022, more than 73,000 human infections have been reported by the CDC from over 100 countries that historically have not reported monkeypox cases. The detection of virus in skin lesions, blood, semen, and saliva of infected patients with monkeypox infections raises the potential for disease transmission via routes that have not been previously documented, including by blood and plasma transfusions. Methods for protecting the blood supply against the threats of newly emerging disease agents exist and include Pathogen Reduction Technologies (PRT) which utilize photochemical treatment processes to inactivate pathogens in blood while preserving the integrity of plasma and cellular components. Such methods have been employed broadly for over 15 years, but effectiveness of these methods under routine use conditions against monkeypox virus has not been reported. Monkeypox virus (strain USA_2003) was used to inoculate plasma and whole blood units that were then treated with riboflavin and UV light (Mirasol Pathogen Reduction Technology System, Terumo BCT, Lakewood, CO). The infectious titers of monkeypox virus in the samples before and after riboflavin + UV treatment were determined by plaque assay on Vero cells. The levels of spiked virus present in whole blood and plasma samples exceeded 103 infectious particles per dose, corresponding to greater than 105 DNA copies per mL. Treatment of whole blood and plasma units under standard operating procedures for the Mirasol PRT System resulted in complete inactivation of infectivity to the limits of detection. This is equivalent to a reduction of ≥ 2.86 +/- 0.73 log10 pfu/mL of infectivity in whole blood and ≥ 3.47 +/-0.19 log10 pfu/mL of infectivity in plasma under standard operating conditions for those products. Based on this data and corresponding studies on infectivity in patients with monkeypox infections, use of Mirasol PRT would be expected to significantly reduce the risk of transfusion transmission of monkeypox.

Efficacy of Short Thermal Treatment Time Against Escherichia coli O157:H7 and Salmonella on the Surface of Fresh Beef

Thermal treatment interventions consistently provide effective pathogen reductions. However, the cost of maintaining high temperature of 95°C in order to raise the surface temperature of carcasses to 82°C is very expensive. Therefore, beef processors need to identify thermal application times and temperatures that optimize the treatment effects with less maintenance cost. The objectives of this study were to determine the efficacy of hot water or steam at 71°C for 6 s and cascade e-ion plasma treatment for 2 s in reducing pathogens on the surface of fresh beef compared to the thermal treatment at 82°C for 15 s. Hot water at 71°C for 6 s reduced Escherichia coli O157:H7 and Salmonella by 2.38 and 2.48 log CFU/cm2, while steam treatment at 71°C for 6 s reduced E. coli O157:H7 and Salmonella by 2.94 and 3.06 log CFU/cm2, respectively. Cascade e-ion plasma treatment for 2 s reduced E. coli O157:H7 on surface of fresh beef by 1.89 log CFU/cm2. The findings indicate that short treatment time with appropriate temperature could serve as an effective carcass intervention to improve the safety of fresh beef.

Responses of roots and rhizosphere of female papaya to the exogenous application of GA3

Exogenous GAs have an indeterminate effect on root development. Our current study used female papaya to reveal how the roots and rhizosphere respond to the exogenous application of GA3 by investigating the transcriptome profile in roots, metabolic profile and microbial community in both roots and rhizosphere of GA3-treated and control female papaya. The results demonstrated that exogenous GA3 treatment enhanced female papaya lateral root development, which gave plants physical advantages of water and nutrient uptake. In addition, it was likely that GA3 spraying in papaya shoot apices increased the level of auxin, which was transported to roots by CpPIN1, where auxin upregulated CpLBD16 and repressed CpBP to promote the lateral root initiation and development. In papaya roots, corresponding transporters (CpTMT3, CpNRT1:2, CpPHT1;4, CpINT2, CpCOPT2, CpABCB11, CpNIP4;1) were upregulated and excretion transporters were downregulated such as CpNAXT1 for water and nutrients uptake with exogenous GA3 application. Moreover, in GA3-treated papaya roots, CpALS3 and CpMYB62 were downregulated, indicating a stronger abiotic resistance to aluminum toxic and phosphate starvation. On the other hand, BRs and JAs, which involve in defense responses, were enriched in the roots and rhizosphere of GA3-treated papayas. The upregulation of the two hormones might result in the reduction of pathogens in roots and rhizosphere such as Colletotrichum and Verticillium. GA3-treated female papaya increased the abundance of beneficial bacteria species including Mycobacterium, Mitsuaria, and Actinophytocola, but decreased that of the genera Candidatus and Bryobacter for that it required less nitrate. Overall, the roots and rhizosphere of female papaya positively respond to exogenous application of GA3 to promote development and stress tolerance. Treatment of female papaya with GA3 might result in the promotion of lateral root formation and development by upregulating CpLBD16 and downregulating CpBP. GA3-treated papaya roots exhibited feedback control of brassinolide and jasmonate signaling in root development and defense. These findings revealed complex response to a growth hormone treatment in papaya roots and rhizosphere and will lead to investigations on the impact of other plant hormones on belowground development in papaya.

Pasteurization of Food and Beverages by High Pressure Processing (HPP) at Room Temperature: Inactivation of Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Salmonella, and Other Microbial Pathogens

Vegetative pathogens actively grow in foods, metabolizing and dividing their cells. They have consequently become a focus of concern for the food industry, food regulators and food control agencies. Although much has been done by the food industry and food regulatory agencies, foodborne outbreaks are still reported globally, causing illnesses, hospitalizations, and in certain cases, deaths, together with product recalls and subsequent economic losses. Major bacterial infections from raw and processed foods are caused by Escherichia coli serotype O157:H7, Salmonella enteritidis, and Listeria monocytogenes. High pressure processing (HPP) (also referred to as high hydrostatic pressure, HHP) is a non-thermal pasteurization technology that relies on very high pressures (400–600 MPa) to inactivate pathogens, instead of heat, thus causing less negative impact in the food nutrients and quality. HPP can be used to preserve foods, instead of chemical food additives. In this study, a review of the effect of HPP treatments on major vegetative bacteria in specific foods was carried out. HPP at 600 MPa, commonly used by the food industry, can achieve the recommended 5–8-log reductions in E. coli, S. enteritidis, L. monocytogenes, and Vibrio. Staphylococcus aureus presented the highest resistance to HPP among the foodborne vegetative pathogens investigated, followed by E. coli. More susceptible L. monocytogenes and Salmonella spp. bacteria were reduced by 6 logs at pressures within 500–600 MPa. Vibrio spp. (e.g., raw oysters), Campylobacter jejuni, Yersinia enterocolitica, Citrobacter freundii and Aeromonas hydrophila generally required lower pressures (300–400 MPa) for inactivation. Bacterial species and strain, as well as the food itself, with a characteristic composition, affect the microbial inactivation. This review demonstrates that HPP is a safe pasteurization technology, which is able to achieve at least 5-log reduction in major food bacterial pathogens, without the application of heat.