Comparative assessment of algaecide performance on freshwater phytoplankton: Understanding differential sensitivities to frame cyanobacteria management.

Cyanobacterial bloom represent a growing threat to global water security. With fast proliferation, they raise great concern due to potential health and socioeconomic concerns. Algaecides are commonly employed as a mitigative measure to suppress and manage cyanobacteria. However, recent research on algaecides has a limited phycological focus, concentrated predominately on cyanobacteria and chlorophytes. Without considering phycological diversity, generalizations crafted from these algaecide comparisons present a biased perpective. To limit the collateral impacts of algaecide interventions on phytoplankton communities it is critical to understand differential phycological sensitivities for establishing optimal dosage and tolerance thresholds. This research attempts to fill this knowledge gap and provide effective guidelines to frame cyanobacterial management. We investigate the effect of two common algaecides, copper sulfate (CuSO4) and hydrogen peroxide (H2O2), on four major phycological divisions (chlorophytes, cyanobacteria, diatoms, and mixotrophs). All phycological divisions exhibited greater sensitivity to copper sulfate, except chlorophytes. Mixotrophs and cyanobacteria displayed the highest sensitivity to both algaecides with the highest to lowest sensitivity being observed as follows: mixotrophs, cyanobacteria, diatoms, and chlorophytes. Our results suggest that H2O2 represents a comparable alternative to CuSO4 for cyanobacterial control. However, some eukaryotic divisions such as mixotrophs and diatoms mirrored cyanobacteria sensitivity, challenging the assumption that H2O2 is a selective cyanocide. Our findings suggest that optimizing algaecide treatments to suppress cyanobacteria while minimizing potential adverse effects on other phycological members is unattainable. An apparent trade-off between effective cyanobacterial management and conserving non-targeted phycological divisions is expected and should be a prime consideration of lake management.

Risk assessment of the antibiotic amoxicillin on non-toxin-producing strains and toxin-producing strains of Microcystis.

Amoxicillin (AMX) is a common antibiotic used to treat a variety of infectious illnesses in humans and animals, including otitis media, tonsillitis, tonsillopharyngitis, laryngitis, and pharyngitis. The drug ends up in the aquatic ecosystems through animal and human excretion and industrial effluents. The ecological consequences of broad-spectrum antibiotics on non-target species like cyanobacteria are causing considerable concern. The danger of amoxicillin to non-toxin-producing and toxin-producing strains of cyanobacteria is poorly understood. The objective of this study was to analyze the risk (RQ) and physiological effects of AMX on Microcystis aeruginosa EAWAG 198 (non-toxin producing = NTP), Microcystis aeruginosa LE3 (toxin-producing = TP), and Microcystis flos aquae UTEX-LB 2677 (toxin-producing = TP). Our study showed differences in the RQ of the drug to the tested organisms - demonstrating < Microcystis flos aquae UTEX-LB 2677 > Microcystis aeruginosa LE3 > Microcystis aeruginosa EAWAG 198. The calculated EC50 values show that AMX was more toxic to the toxin-producing strains than the non-toxin-producing strains. Amoxicillin led to significant (p < 0.05) growth inhibition and chlorophyll-a content of the exposed cultures. The observed increase in the concentration of intracellular hydrogen peroxide (H2O2) of the exposed cultures at 96 h was significant (p < 0.05), demonstrating that the expressed oxidative stress patterns observed during the study were due to AMX. The current study shows significant variation (p < 0.05) in melondialdehyde (MDA) content and the antioxidant enzymes - glutathione-S-transferase (GST) and peroxidase (POD).

Assessment of three alternative methods for bacterial disinfection of hatching eggs in comparison with conventional approach in commercial broiler hatcheries.

The disinfection of commercial hatching eggs before incubation is a common strategy to reduce potential vertical transmission of bacterial and fungal infections from the eggshell to one-day-old chicks that may prevail in poultry products and eventually reach the end consumer. The present investigation focuses on the parallel testing and application of four different disinfection methods (conventional and alternative) under commercial hatchery conditions against natural eggshell bacterial contamination. Hatching eggs from two ROSS 308 broiler breeder flocks were selected and divided into six different groups: two groups were not disinfected and served as negative controls, and four were independently disinfected following product specifications and protocols. From each group, a sample of 100 hatching eggs was selected for bacterial re-isolation, utilizing a modified shell rinse method. Colony-forming units (cfu) from the shell rinse suspensions were determined and analyzed to establish cfu values for each tested egg. These values were analyzed to determine the bacterial disinfection capacity of the four disinfection methods under commercial hatchery conditions. The tested methods were hydrogen peroxide + alcohol, peracetic acid, low energy electron beam, and the gold standard in practice: formaldehyde. Among these methods, formaldehyde, peracetic acid, and low energy electron beam showed a significant difference when compared to the non-disinfected groups whereas hydrogen peroxide + alcohol did not. The bacterial disinfection capacity of the tested methods was compared as well to the gold standard method formaldehyde fumigation and only low energy electron beam achieved similar disinfection levels as formaldehyde. According to our data, three methods significantly reduce the bacterial load on the eggshell of hatching eggs under commercial hatching conditions, including potential alternative methods such as low energy electron beam that perform similar to the gold standard in practice.

Safety Assessment of the Modified Lactoperoxidase System-In Vitro Studies on Human Gingival Fibroblasts.

One strategy in caries prevention is to inhibit the formation of cariogenic biofilms. Attempts are being made to develop oral hygiene products enriched with various antimicrobial agents. One of them is lactoperoxidase-an enzyme that can oxidise (pseudo)halide ions to reactive products with antimicrobial activity. Currently, commercially available products utilise thiocyanate as a substrate; however, several alternatives that are oxidised to products with greater antimicrobial potential have been found. In this study, toxicity against human gingival fibroblasts of the lactoperoxidase system was evaluated using four different (pseudo)halide substrate systems-thiocyanate, iodide, selenocyanate, and a mixture of thiocyanate and iodide. For this purpose, cells were treated with the systems and then apoptosis, cell cycle, intracellular glutathione concentration, and mitochondrial superoxide production were assessed. The results showed that each system, after generating 250 µM of the product, inhibited cell divisions, increased apoptosis, and increased the percentage of dead cells. It was concluded that the mechanism of the observed phenomena was not related to increased superoxide production or the depletion of glutathione concentration. These findings emphasised the need for the further in vitro and in vivo toxicity investigation of the modified lactoperoxidase system to assess its safety and the possibility of use in oral hygiene products.

Application of a Dual-Probe Coloading Nanodetection System in the Process Monitoring and Efficacy Assessment of Photodynamic Therapy: An In Vitro Study.

The oxygen-consuming property of photodynamic therapy (PDT) affects its effects and aggravates tumor hypoxia, thus upregulating the vascular endothelial growth factor (VEGF) to exacerbate tumor metastasis and lead to treatment failure. Therefore, it is necessary to monitor the dynamic changes in the factors related to PDT and tumor development trends in real time, thus helping to improve PDT efficiency. This study fabricated a fluorescent probe, TPE-2HPro, and a fluorescein-labeled aptamer probe, FAM-AptamerVEGF, to detect hydrogen peroxide (H2O2) and VEGF through the photoinduced electron-transfer effect and the specific affinity of the aptamer to VEGF, respectively. The two probes were loaded into the inner pores and absorbed on the surface of polydopamine coating-wrapped mesoporous silica nanoparticles (MSN@PDA) to construct the dual-probe-loaded system, MSNTH@PDAApt, which was kept stable in fetal bovine serum (FBS) solution and achieved pH-responsive release behavior, thus helping to increase the accumulation of the two probes in tumor cells. The dichloroacetic acid-mediated in vitro antitumor tests showed that the changing trends of H2O2 and VEGF levels were consistent with the results of related mechanism studies and could be monitored by MSNTH@PDAApt. The in vitro chlorin e6 (Ce6)-mediated PDT treatment demonstrated that when the illumination condition was 650 nm, 50 mW/cm2 for 10 min, cells were more inclined to metastasis and invasion rather than death due to a substantial increase in VEGF expression at the low Ce6 concentrations. With the increase of the Ce6 concentration, the growth of the H2O2 level gradually exceeded that of VEGF, and the reactive oxygen species (ROS)-mediated cell death dominated when the Ce6 concentration was about 2 times its IC50 values. Besides, hypoxia also affected the H2O2 and VEGF changes. These results demonstrated that MSNTH@PDAApt could precisely monitor and assess the tumor development trends during PDT treatment, thus helping improve the treatment effect.

Curcumin/H2O2 photodynamically activated: an antimicrobial time-response assessment against an MDR strain of Candida albicans.

OBJECTIVE: Human candidiasis is typically treated with antifungal drugs, but the rise of drug-resistant strains of Candida spp. poses a serious problem, making treatment difficult. At the same time, photodynamic therapy (PDT) has drawn increasing attention from researchers for its potential to effectively inhibit multidrug-resistant pathogenic fungi and for its low tendency to induce drug resistance. This study's goal was to examine how a multidrug-resistant oral isolate of Candida albicans responded to a PDT that used a curcumin/H202 formulation as a photosensitizer and was exposed to various light sources. MATERIALS AND METHODS: A commercial product containing curcumin/H2O2 3% was used as a photosensitizer and evaluated in a PDT treatment that can use two different light sources: traditional irradiation with 7 W light at λ = 460 nm or a new, never evaluated, polarized light source of 25 W with a wavelength range of λ = 380-3,400 nm. The antimicrobial activity of these procedures was assessed on a clinical oral isolate of Candida albicans, in terms of agar susceptibility test, growth curve behavior, and biofilm inhibition. RESULTS: Both light sources were able to activate the photosensitizer formulation, suggesting a fungistatic activity vs. this C. albicans MDR strain. An interesting difference was observed in the cell-generation-time (CGTOD) after PDT treatment, where the polarized light was more active compared to the source of 460 nm. In fact, CGTOD was 16 and 8 hours, respectively. CONCLUSIONS: The PDT evaluated here presented an inhibition window time, a crucial point for clinicians, who could activate an additional prophylactic treatment to resolve the clinical management of Candida infections in the oral cavity.

Comparative assessment of the efficacy of low concentration bleaching agents using quantitative light induced fluorescence in removing stains: An In vitro study.

Background: Tooth discoloration has become a common esthetic problem in recent years. Removal of stains by bleaching is well-documented. Low concentration home bleaching products are available in market in different forms and concentrations. Aim: The aim of this study is to evaluate and compare the efficacy of low concentration commercially available home bleaching products (whitening strip, gel, and mouthwash) in removing stains and whitening the tooth using clinical and digital methods. Materials and Methods: Sixty permanent enamel samples mounted in an acrylic block were artificially stained and randomly divided into four groups. Negative control, 15 % Carbamide peroxide gel group, 2% Hydrogen 16 peroxide mouthwash group and 6% Hydrogen peroxide strip group respectively. The samples were bleached with respective agents according to the manufacturer's instructions. The efficacy on 7th and 14th day was evaluated clinically (SGU change), photographically (ΔE), and using quantitative light-induced fluorescence (ΔF). The data were analyzed using paired t-test and analysis of variance. Results: Postbleaching, 6% hydrogen peroxide strips and 15% carbamide peroxide gel showed maximum improvement (ΔΔF - 15.73 and 11.89, ΔE - 19.8 and 18.9, respectively) when compared to 2% hydrogen peroxide mouthwash and negative control group (ΔΔF - 9.68 and 6.59, ΔE - 15.04 and 9.44, respectively). The difference was statistically significant (P = 0.001). Conclusion: 6% hydrogen peroxide strips and 15% carbamide peroxide gel showed maximum improvement in stain removal and tooth whitening however, the strips showed better efficacy than the gel. Strips have the added advantage of lesser contact period, less salivary dilution, and no gingival contact. Therefore, strips can be a better alternative for gels and mouthwashes.

Efficacy of hydrogen peroxide wipes for decontamination of AZD1222 adenovirus COVID-19 vaccine strain on pharmaceutical industry materials.

This study aimed to evaluate the performance of accelerated hydrogen peroxide® wipes (HPW) for decontamination of the chimpanzee adenovirus AZD1222 vaccine strain used in the production of recombinant COVID-19 vaccine in a pharmaceutical industry. Two matrices were tested on stainless-steel (SS) and low-density-polyethylene (LDP) surfaces: formulated recombinant COVID-19 vaccine (FCV) and active pharmaceutical ingredient (API). The samples were spiked, dried and the initial inoculum, possible residue effect (RE) and titre reduction after disinfection with HPW were determined. No RE was observed. The disinfection procedure with HPW resulted in complete decontamination the of AZD1222 adenovirus strain in FCV (≥7·46 and ≥7·49 log10 infectious unit [IFU] ml-1 for SS and LDP carriers respectively) and API (≥8·79 and ≥8·78 log10 IFU ml-1 for SS and LDP carriers respectively). In conclusion, virucidal activity of HPW was satisfactory against the AZD1222 adenovirus strain and can be a good option for disinfection processes of SS and LPD surfaces in pharmaceutical industry facilities during recombinant COVID-19 vaccine production. This procedure is simple and can be also applied on safety unit cabins and sampling bags made of LDP as well.

Efficacy assessment of different time cycles of nebulized hydrogen peroxide against bacterial and yeast biofilms.

BACKGROUND: The prevention of healthcare-associated infections requires continuous effort. In order to achieve better practical results, the control of environmental microbial biofilms with effective disinfection strategies should be addressed. AIM: To test the efficacy of different time cycles of nebulized hydrogen peroxide (H2O2) against bacterial and yeast dry biofilms. METHODS: The efficacies of a standard cycle (SC) and a fast cycle (FC) of nebulized H2O2 were compared. Microbial biofilms were grown on different material coupons. The metabolic activity of biofilms was determined by XTT assay, and the total biomass of biofilms was determined by crystal violet assay. FINDINGS: Regarding the efficacy of nebulized H2O2 against biofilms, the mean reduction in metabolic activity for the SC was 55.2% [standard deviation (SD) 19.4%], compared with 50.4% (SD 17.7%) for the FC. The mean reduction in total biomass for the SC was 45.5% (SD 22.7%), compared with 46.7% (SD 21.7%) for the FC. No significant differences were found between the tested cycles and materials. CONCLUSION: H2O2 nebulization was found to exhibit good efficacy against healthcare-associated microbial dry biofilms. Moreover, similar efficacies were found between the SC and the FC.

Elevated atmospheric CO2 enhances the phytoremediation efficiency of tall fescue (Festuca arundinacea) in Cd-polluted soil.

With the economic development of society, concentrations of atmospheric CO2 and heavy metals in soils have been increasing. The physiological responses of plants to the interaction between soil pollution and climatic change need to be understood. Pot experiments were designed to assess variations in Festuca arundinacea dry weight, leaf type, chlorophyll content, antioxidase activities, and Cd accumulation ability, under different atmospheric CO2 treatments. The results showed that the total dry weights increased with increasing CO2, and Cd concentrations in falling leaf tissues increased with raised atmospheric CO2, before reaching a peak at 600 ppm, above which they remained constant. Compared with the control (400 ppm), 600, 650, and 700 ppm CO2 treatments increased the proportions of the falling tissues by 1.7%, 3.3%, and 4.5%, respectively. Antioxidant enzyme activities in plant leaves increased with increasing atmospheric CO2 levels. The concentration of H2O2 in leaf tissues increased with increasing CO2, reaching a peak at 600 ppm, and then decreased significantly as the CO2 content increased further, to 700 ppm. The results in this study suggest that F. arundinacea could be regarded as a potential candidate for phytoremediation of Cd-polluted soil; especially if senescent and dead leaf tissues could be harvested, and that raised atmospheric CO2 levels could improve its soil remediation efficiency.Novelty statement Extrapolation of results from experiments of environmental impacts in greenhouse to real scale field requires to be considered cautiously. External factors such as water, temperature, humidity, and pollution are variable in real field. Plants will face a lot of beneficial or detrimental conditions which will influence the magnitude of the results. However, the elevation of CO2 is an inevitable phenomenon in future. Therefore, findings from experiments under artificial conditions are sometime a good choice to obtain knowledge about elevated CO2 related impacts on phytoremediation efficiency of a specific plant. The final goal of this work is to find a suitable CO2 fumigation strategy optimized for soil remediation. We report on that elevated atmospheric CO2 can increase the phytoremediation efficiency of Festuca arundinacea for Cd. This is significant because the combined influences of elevated atmospheric CO2 and metal pollution in terms of biomass yield, pollutant uptake, and phytoremediation efficiency would be more complex than the effects of each individual factor.

Evaluation of hydrogen peroxide efficacy against AZD1222 chimpanzee adenovirus strain in the recombinant COVID-19 vaccine for application in cleaning validation in a pharmaceutical manufacturing industry.

This study aimed to evaluate the performance of hydrogen peroxide vapour (HPV) to inactivate the chimpanzee adenovirus AZD1222 vaccine strain used in the production of recombinant COVID-19 vaccine for application in cleaning validation in pharmaceutical industries production areas. Two matrixes were tested: formulated recombinant COVID-19 vaccine (FCV) and active pharmaceutical ingredient (API). The samples were dried on stainless steel and exposed to HPV in an isolator. One biological indicator with population >106 Geobacillus stearothermophilus spores was used to validate the HPV decontamination cycle as standard. HPV exposure resulted in complete virus inactivation in FVC (≥5·03 log10 ) and API (≥6·40 log10 ), showing HPV efficacy for reducing chimpanzee adenovirus AZD1222 vaccine strain. However, the optimum concentration and contact time will vary depending on the type of application. Future decontamination studies scaling up the process to the recombinant COVID-19 vaccine manufacturing areas are necessary to evaluate if the HPV will have the same or better virucidal effectivity in each specific production area. In conclusion, HPV showed efficacy for reducing AZD1222 chimpanzee adenovirus strain and can be a good choice for pharmaceutical industries facilities disinfection during recombinant COVID-19 vaccine production.

In vitro assessment of synergistic effects in combinations of a temoporfin-based photodynamic therapy with glutathione peroxidase 1 inhibitors.

BACKGROUND: Due to an increased elimination of reactive oxygen species (ROS), in particular hydrogen peroxide (H2O2), overexpression of glutathione peroxidase 1 (GPX1) can lead to an attenuation of apoptosis and development of resistance in cancer cells, thereby promoting tumor cell survival. Consequently, GPX1 inhibitors have the potential to be used in cancer therapy as they support oxidative stress in cancer cells. Similarly, photodynamic therapy (PDT) induces oxidative stress in cancer cells by the formation of ROS upon illumination. Thus, both methods of treatment might act in synergy when used in combination. METHODS: To investigate this hypothesis, combinations of the known GPX1 inhibitors 9-chloro-6-ethyl-6H-[1,2,3,4,5]pentathiepino[6,7-b]indole (CEPI) or mercaptosuccinic acid (MSA) with PDT induced by the photosensitizer (PS) temoporfin (5,10,15,20-tetra(m-hydroxyphenyl)chlorin, mTHPC) were studied in vitro. This new combinatory approach was intended to accumulate ROS formed during PDT via blockage of GPX1-catalyzed H2O2 degradation, and thus to enhance PDT-induced phototoxicity. Five human cancer cell lines from tumor origins treatable with PDT were utilized to investigate ROS generation, apoptosis induction, and cell cycle distribution. RESULTS: Synergy was identified with both GPX1 inhibitors, but not in all cell lines. ROS levels were increased after combined treatment with mTHPC and CEPI, but not MSA, in some cell lines, indicating that oxidative stress and ROS accumulation were enhanced by CEPI. Surprisingly, enhanced apoptosis induction was also observed with MSA afterwards, suggesting that other pathways contributed to the initiation of apoptosis. Cell cycle analysis confirmed apoptosis induction via the detection of DNA fragmentation. CONCLUSION: A combination of GPX1 inhibitors with mTHPC-PDT has the potential to generate synergistic effects and to increase overall phototoxicity, but the success of this combination approach was dependent on cancer type, and even antagonistic effects can occur.

Systematic review on use, cost and clinical efficacy of automated decontamination devices.

BACKGROUND: More evidence is emerging on the role of surface decontamination for reducing hospital-acquired infection (HAI). Timely and adequate removal of environmental pathogens leads to measurable clinical benefit in both routine and outbreak situations. OBJECTIVES: This systematic review aimed to evaluate published studies describing the effect of automated technologies delivering hydrogen peroxide (H202) or ultra-violet (UV) light on HAI rates. METHODS: A systematic review was performed using relevant search terms. Databases were scanned from January 2005 to March 2020 for studies reporting clinical outcome after use of automated devices on healthcare surfaces. Information collected included device type, overall findings; hospital and ward data; study location, length and size; antimicrobial consumption; domestic monitoring; and infection control interventions. Study sponsorship and duplicate publications were also noted. RESULTS: While there are clear benefits from non-touch devices in vitro, we found insufficient objective assessment of patient outcome due to the before-and-after nature of 36 of 43 (84%) studies. Of 43 studies, 20 (47%) used hydrogen peroxide (14 for outbreaks) and 23 (53%) used UV technology (none for outbreaks). The most popular pathogen targeted, either alone or in combination with others, was Clostridium difficile (27 of 43 studies: 63%), followed by methicillin-resistant Staphylococcus aureus (MRSA) (16 of 43: 37%). Many owed funding and/or personnel to industry sponsorship (28 of 43: 65%) and most were confounded by concurrent infection control, antimicrobial stewardship and/or cleaning audit initiatives. Few contained data on device costs and rarely on comparable costs (1 of 43: 2%). There were expected relationships between the country hosting the study and location of device companies. None mentioned the potential for environmental damage, including effects on microbial survivors. CONCLUSION: There were mixed results for patient benefit from this review of automated devices using H202 or UV for surface decontamination. Most non-outbreak studies lacked an appropriate control group and were potentially compromised by industry sponsorship. Concern over HAI encourages delivery of powerful disinfectants for eliminating pathogens without appreciating toxicity or cost benefit. Routine use of these devices requires justification from standardized and controlled studies to understand how best to manage contaminated healthcare environments.

Decontaminating N95 respirators during the COVID-19 pandemic: simple and practical approaches to increase decontamination capacity, speed, safety and ease of use.

BACKGROUND: The COVID-19 pandemic has caused a severe shortage of personal protective equipment (PPE), especially N95 respirators. Efficient, effective and economically feasible methods for large-scale PPE decontamination are urgently needed. AIMS: (1) to develop protocols for effectively decontaminating PPE using vaporized hydrogen peroxide (VHP); (2) to develop novel approaches that decrease set-up and take-down time while also increasing decontamination capacity; (3) to test decontamination efficiency for N95 respirators heavily contaminated by make-up or moisturizers. METHODS: We converted a decommissioned Biosafety Level 3 laboratory into a facility that could be used to decontaminate N95 respirators. N95 respirators were hung on metal racks, stacked in piles, placed in paper bags or covered with make-up or moisturizer. A VHP® VICTORY™ unit from STERIS was used to inject VHP into the facility. Biological and chemical indicators were used to validate the decontamination process. FINDINGS: N95 respirators individually hung on metal racks were successfully decontaminated using VHP. N95 respirators were also successfully decontaminated when placed in closed paper bags or if stacked in piles of up to six. Stacking reduced the time needed to arrange N95 respirators for decontamination by approximately two-thirds while almost tripling facility capacity. Make-up and moisturizer creams did not interfere with the decontamination process. CONCLUSIONS: Respirator stacking can reduce the hands-on time and increase decontamination capacity. When personalization is needed, respirators can be decontaminated in labelled paper bags. Make up or moisturizers do not appear to interfere with VHP decontamination.

Hepatoprotective activity assessment of amino acids derivatives of picroside I and II.

Picrorhiza kurroa has a long medicinal history as a traditional medicinal plant in China and India that is widely used in clinical treatments. It is a common treatment for liver diseases, fever, diarrhoea, indigestion, and some other diseases. Modern pharmacological studies proved that P. kurroa rhizomes have high levels of picroside I and II, which were identified as main constituents with anti-inflammatory and hepatoprotective activities. In our study, we used picroside I and II as the lead compounds to generate derivatives by reactions with Boc-valine or Boc-proline, which underwent dehydration and condensation with the hydroxyl groups in the lead compounds in the presence of coupling reagent N,N'-dicyclohexylcarbodiimide. We synthesized 11 derivatives and examined their hepatoprotective effects in vitro by assessing the proliferation rates of H2 O2 -exposed HepG2 cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. We found that some derivatives promoted higher proliferation rates in HepG2 cells than the natural compounds before derivatization, suggesting that those derivatives possessed an improved hepatoprotective capacity. The novel derivatization strategy for picrosides had the additional benefit that the esterification of their hydroxyl groups created derivatives not only with increased stability but also with improved pharmacokinetic properties and potentially prolonged half-life.

Assessment of Stabilized Hydrogen Peroxide for Use in Reducing Campylobacter Levels and Prevalence on Broiler Chicken Wings.

ABSTRACT: Poultry processing establishments use antimicrobial aids on broiler parts to minimize Campylobacter contamination. A silver-stabilized hydrogen peroxide (SHP) product was assessed for use as an antimicrobial processing aid. In a series of experiments, wing segments with skin were inoculated with 103 to 107 cells of Campylobacter coli, followed by treatment with SHP at 15,000 or 30,000 mg/L, peroxyacetic acid (PAA) at 300 or 3,000 mg/L (parts per million), or water. Each treatment was applied by either dip or spray. Rinsates from each wing segment were analyzed for direct counts and prevalence of Campylobacter. Treatment with SHP or PAA significantly reduced Campylobacter levels compared with water controls by up to 2.22 log CFU/mL. At high inoculum levels (106 to 107), SHP and PAA applied by dip had up to 1.27 log CFU/mL further reductions of Campylobacter levels compared with spray-treated wing segments. Additionally, wing drumettes were observed to retain higher levels and prevalence of Campylobacter recovery compared with wing flats at a low inoculation level (103). The results indicated that there was no carryover effect of SHP (same day versus 24 h) and dip treatment with SHP or PAA decreased Campylobacter recovery on broiler chicken wing segments compared with a water control. Although a 2-log reduction was modest, SHP had similar efficacy as the commonly used processing aid PAA. SHP shows potential for further investigation as an antimicrobial processing aid for use on poultry parts.

Diversity Assessment of Toxic Cyanobacterial Blooms during Oxidation.

Fresh-water sources of drinking water are experiencing toxic cyanobacterial blooms more frequently. Chemical oxidation is a common approach to treat cyanobacteria and their toxins. This study systematically investigates the bacterial/cyanobacterial community following chemical oxidation (Cl2, KMnO4, O3, H2O2) using high throughput sequencing. Raw water results from high throughput sequencing show that Proteobacteria, Actinobacteria, Cyanobacteria and Bacteroidetes were the most abundant phyla. Dolichospermum, Synechococcus, Microcystis and Nostoc were the most dominant genera. In terms of species, Dolichospermum sp.90 and Microcystis aeruginosa were the most abundant species at the beginning and end of the sampling, respectively. A comparison between the results of high throughput sequencing and taxonomic cell counts highlighted the robustness of high throughput sequencing to thoroughly reveal a wide diversity of bacterial and cyanobacterial communities. Principal component analysis of the oxidation samples results showed a progressive shift in the composition of bacterial/cyanobacterial communities following soft-chlorination with increasing common exposure units (CTs) (0-3.8 mg·min/L). Close cyanobacterial community composition (Dolichospermum dominant genus) was observed following low chlorine and mid-KMnO4 (287.7 mg·min/L) exposure. Our results showed that some toxin producing species may persist after oxidation whether they were dominant species or not. Relative persistence of Dolichospermum sp.90 was observed following soft-chlorination (0.2-0.6 mg/L) and permanganate (5 mg/L) oxidation with increasing oxidant exposure. Pre-oxidation using H2O2 (10 mg/L and one day contact time) caused a clear decrease in the relative abundance of all the taxa and some species including the toxin producing taxa. These observations suggest selectivity of H2O2 to provide an efficient barrier against toxin producing cyanobacteria entering a water treatment plant.

A Medium-Throughput System for In Vitro Oxidative Stress Assessment in IPEC-J2 Cells.

The feed industry continuously seeks new molecules with antioxidant capacity since oxidative stress plays a key role in intestinal health. To improve screening of new antioxidants, this study aims to set up an assay to assess oxidative stress in the porcine small intestinal epithelial cell line IPEC-J2 using plate-reader-based analysis of fluorescence. Two oxidants, H2O2 and menadione, were tested at 1, 2 and 3 mM and 100, 200 and 300 µM, respectively. Trolox (2 mM) was used as the reference antioxidant and the probe CM-H2DCFDA was used to indicate intracellular oxidative stress. Cell culture, reactive oxygen species (ROS) production and assessment conditions were optimized to detect a significant ROS accumulation that could be counteracted by pre-incubation with trolox. Menadione (200 µM) reproducibly increased ROS levels, H2O2 failed to do so. Trolox significantly decreased intracellular ROS levels in menadione (200 µM)-exposed cells in a consistent way. The system was further used to screen different concentrations of the commercially available antioxidant ELIFE®. Concentrations between 100 and 200 ppm protected best against intracellular ROS accumulation. In conclusion, the combination of CM-H2DCFDA fluorescence analysis by a plate-reader, trolox as a reference antioxidant and 200 µM of menadione as a stressor agent, provides a replicable and reliable medium-throughput setup for the evaluation of intracellular oxidative stress in IPEC-J2 cells.

A break-even analysis of benzoyl peroxide and hydrogen peroxide for infection prevention in shoulder arthroplasty.

BACKGROUND: Newer strategies to decolonize the shoulder of Cutibacterium acnes may hold promise in minimizing the occurrence of infections after shoulder arthroplasty, but little is known about their cost-effectiveness. Break-even models can determine the economic viability of interventions in settings with low outcome event rates that would realistically preclude a randomized clinical trial. We used such modeling to determine the economic viability of benzoyl peroxide and hydrogen peroxide for infection prevention in shoulder arthroplasty. METHODS: Skin decolonization protocol costs ($11.76 for benzoyl peroxide; $0.96 for hydrogen peroxide), baseline infection rates for shoulder arthroplasty (0.70%), and infection-related care costs ($50,230) were derived from institutional records and the literature. A break-even equation incorporating these variables was developed to determine the absolute risk reduction (ARR) in the infection rate to make prophylactic use economically justified. The number needed to treat was calculated from the ARR. RESULTS: Topical benzoyl peroxide is considered economically justified if it prevents at least 1 infection out of 4348 shoulder arthroplasties (ARR = 0.023%). Hydrogen peroxide is economically justified if it prevents at least 1 infection out of 50,000 cases (ARR = 0.002%). These protocols remained economically viable at varying unit costs, initial infection rates, and infection-related care costs. CONCLUSIONS: The use of topical benzoyl peroxide and skin preparations with hydrogen peroxide are highly economically justified practices for infection prevention in shoulder arthroplasty. Efforts to determine drawbacks of routine skin decolonization strategies are warranted as they may change the value analysis.

Next-generation multiparameter flow cytometry assay improves the assessment of oxidative stress in probiotics.

Stability of probiotic products' potency throughout shelf life is essential to ensure systematic delivery of the dosages required to provide clinically-proven health benefits. Due to the oxygen sensitivity of gut-derived microorganisms, methods for the rapid and accurate monitoring of oxidative stress in probiotics are greatly needed as they can be instrumental to both bioprocess optimization and quality control. This study introduces a next-generation flow cytometry method multiplexing the CellROX® Green and Propidium Iodide probes for the simultaneous measurement of free total reactive oxygen species (ROS) and membrane integrity, respectively. The multiparameter method was compared to the single-parameter assays, measuring either ROS or membrane integrity, for the ability to evaluate the fitness of Lactobacillus rhamnosus GG (LGG) after freeze drying, spray drying and H2O2-mediated oxidative stress. Each stand-alone assay detected only three cell populations, showing either differential membrane integrity (Syto 24+/PI-, Syto 24+/PI+, Syto 24-/PI+) or ROS levels (ROS-, low-ROS, high-ROS), and no correlation could be drawn between these groups. Conversely, the multiparameter method detected up to five physiologically distinct cell populations and allowed the integrated assessment of their membrane integrity and oxidative stress. It also revealed a much larger fitness heterogeneity in LGG as each group of low-ROS and high-ROS cells was found to be formed by a healthier population with an intact membrane (L-ROS/PI-, H-ROS/PI-) and a population with damaged membrane (L-ROS/PI+, H-ROS/PI+). As the CRG probe only detects free unreacted ROS, these populations are suggested to reflect the dynamic lifecycle of ROS formation, accumulation and reactive depletion leading to oxidative damage of macromolecules and consequent cell death. With the stand-alone CRG assay being unable to detect ROS lifecycle, the multiparameter method here presented delivers a superior profiling of the heterogeneity generated by oxidative stress in bacteria and enables a more correct interpretation of CRG fluorescence data. We provide recent examples from literature where the use of a single-parameter fluorescence approach may have led to misinterpret oxidative stress data and eventually draw erroneous conclusions.