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Microorganisms May 2024Environmental decontamination and water disinfection practices are hallmarks of disease prevention and control in agricultural and public health settings. Informed...
Flow Cytometry Coupled with Resuscitation Assays As a High-Resolution Tool to Inform Environmental Management and Disinfection of Settings Affected by Tuberculous Mycobacteria.
Environmental decontamination and water disinfection practices are hallmarks of disease prevention and control in agricultural and public health settings. Informed fit-to-purpose biocontainment is thus dependent on methodologies accurately assessing microbial burden and viability. Also, rigorous evaluation of the efficacy of biocontrol measures implies monitoring microbial inactivation after decontamination/disinfection procedures. In this study, we used flow cytometry coupled with a resuscitation protocol to monitor the metabolic inactivation of bacteria capable of entering non-cultivable states, after the application of a chlorine-based water disinfectant. For this purpose, we used BCG as a model of slow-growing bacteria able to enter dormancy and representing a multi-host pathogen in a zoonotic disease system-animal tuberculosis-thriving both across temperate and semi-arid regions and involving environmental contamination. The biocide activity of a commercial sodium dichloroisocyanurate (NaDCC) disinfectant against BCG was evaluated through mock environmental matrix tests. Using the manufacturer-recommended dosage of NaDCC, BCG cells were apparently inactivated after 24 h upon exposure. However, we show via flow cytometry that, upon exposure to optimal growth conditions, mycobacterial cells were able to regain metabolic activity shortly after, highlighting a sublethal effect of NaDCC at the recommended commercial dosage due to reversible BCG cell damage. In contrast, increasing twice the disinfectant dosage completely inactivated BCG cells after 24 h of exposure, with full irreversible loss of metabolic activity. Methodological workflows based on conventional culture or PCR would have missed the detection of these dormant subpopulations that were in fact able to resume growth when following the recommendations of a commercial disinfectant. This study highlights the superior, high-resolution value of single-cell approaches, such as flow cytometry, to accurately assess the activity of biocides against metabolically heterogeneous and dormant pathogenic bacteria with environmental cycles, supporting data-driven prioritization of environmental management and disinfection options in contaminated vulnerable settings.
PubMed: 38930449
DOI: 10.3390/microorganisms12061068 -
Animals : An Open Access Journal From... Jun 2024The aim of this review was to analyse the health problem of lameness in dairy cows by assessing the health and economic losses. This review also presents in detail the... (Review)
Review
The aim of this review was to analyse the health problem of lameness in dairy cows by assessing the health and economic losses. This review also presents in detail the etiopathogenesis of lameness in dairy cattle and examples of its treatment and prevention. This work is based on a review of available publications. In selecting articles for the manuscript, the authors focused on issues observed in cattle herds during their clinical work. Lameness in dairy cattle is a serious health and economic problem around the world. Production losses result from reduced milk yield, reduced feed intake, reproductive disorders, treatment costs, and costs associated with early culling. A significant difficulty in the control and treatment of lameness is the multifactorial nature of the disease; causes may be individual or species-specific and may be associated with the environment, nutrition, or the presence of concomitant diseases. An important role is ascribed to infectious agents of both systemic and local infections, which can cause problems with movement in animals. It is also worth noting the long treatment process, which can last up to several months, thus significantly affecting yield and production. Given the high economic losses resulting from lameness in dairy cows, reaching even >40% (depending on the scale of production), there seems to be a need to implement extensive preventive measures to reduce the occurrence of limb infections in animals. The most important effective preventive measures to reduce the occurrence of limb diseases with symptoms of lameness are periodic hoof examinations and correction, nutritional control, and bathing with disinfectants. A clean and dry environment for cows should also be a priority.
PubMed: 38929454
DOI: 10.3390/ani14121836 -
Animals : An Open Access Journal From... Jun 2024This paper describes the selection and validation of supporting measures (SMs) aimed at enhancing biosecurity compliance within Italian poultry farms. A tailored...
This paper describes the selection and validation of supporting measures (SMs) aimed at enhancing biosecurity compliance within Italian poultry farms. A tailored methodology, based on a stakeholders' survey involving farmers and advisors, included a virtual farm tour, group discussion, and farmer coaching. Virtual farm tours and group discussions were delivered during two meetings targeting meat and egg production stakeholders, separately. Coaching was validated in 26 pilot farms (PFs) by assessing farmers' attitudes towards change (i.e., ADKAR) and farms' biosecurity score (i.e., Biocheck.Ugent) before and after a minimum six-month period. A total of 20 out of 26 farmers agreed to implement at least one action plan (AP). Full implementation of the agreed APs was observed in ten farms, while others only partially implemented ( = 7) or did not implement ( = 3) the improvement. Most APs focused on enhancing house hygiene locks ( = 7), followed by bacterial auto-control after cleaning and disinfection ( = 4). Scoring tools indicated minimal or no variations in farmers' attitudes towards change and farm biosecurity. Virtual farm tours and group discussions were found to be effective in fostering interaction and facilitating the exchange of experiences and knowledge among farmers and stakeholders of poultry production. Coaching indicated that farmers might prefer implementing minor changes possibly influenced by time and cost constraints associated with structural interventions. These limitations could have also impacted the scores of the farmer/farm. The findings of this study provide a foundation for further application of SMs to improve biosecurity in Italian poultry farms.
PubMed: 38929353
DOI: 10.3390/ani14121734 -
Antioxidants (Basel, Switzerland) May 2024Curcumin (Curcumin) belongs to the polyphenol family. It is extracted by drying the root of a plant of Asian origin, belonging to the family. The best-known species is... (Review)
Review
Curcumin (Curcumin) belongs to the polyphenol family. It is extracted by drying the root of a plant of Asian origin, belonging to the family. The best-known species is . Curcumin has been recognized as having great therapeutic powers since ancient times. Studies on curcumin have since confirmed its powerful antioxidant properties, preventing both the formation of free radicals and their neutralization, having anti-inflammatory, antibacterial, immunological, and neuroprotective properties, as well as being a regulator of the intestinal microbiota with beneficial effects on the clinical manifestations of metabolic syndrome. Our study aimed to highlight how all these therapeutic aspects could benefit oral health, both preventing and improving the course of pathological processes. The effect of mouthwashes, and curcumin-based gels on the regulation of bacterial plaque and in the control of gingivitis, was largely comparable to that of using 0.20% chlorhexidine, with fewer side effects. Being a highly hydrophobic substance, it has a high permeability to cross the cell membrane. Bioavailability increases when combined with liposoluble substances (e.g., olive oil) and piperine, which improves absorption. Curcumin also has a negligible degree of toxicity, making it an excellent alternative to the use of gold standard products for oral disinfection.
PubMed: 38929099
DOI: 10.3390/antiox13060660 -
International Journal of Environmental... May 2024Healthcare-acquired infections are a major problem in healthcare facility settings around the world. The Democratic Republic of the Congo (DRC) has over 2 million... (Randomized Controlled Trial)
Randomized Controlled Trial
Effect of a Water, Sanitation, and Hygiene Program on Handwashing with a Cleansing Agent among Diarrhea Patients and Attendants in Healthcare Facilities in the Democratic Republic of the Congo: A Randomized Pilot of the PICHA7 Program.
Healthcare-acquired infections are a major problem in healthcare facility settings around the world. The Democratic Republic of the Congo (DRC) has over 2 million diarrhea patients hospitalized each year. These healthcare settings become high-risk environments for spreading diarrheal illnesses such as cholera. The objective of the Preventative Intervention for Cholera for 7 Days (PICHA7) program is to develop evidence-based water, sanitation, and hygiene (WASH) interventions to reduce cholera and other severe diarrheal diseases in the DRC. The study objective was to evaluate the effectiveness of PICHA7 program delivery in increasing handwashing with a cleansing agent at stool/vomit- and food-related events in a healthcare facility setting among diarrhea patients and patient attendants. A pilot of the PICHA7 program was conducted among 284 participants in 27 healthcare facilities from March 2020 to November 2021 in urban Bukavu in the South Kivu Province of the DRC. The standard arm received the standard message provided in the DRC to diarrhea patients on the use of oral rehydration solution and a basic WASH message at healthcare facility discharge. The PICHA7 arm received the PICHA7 WASH pictorial module delivered by a health promoter focused on handwashing with a cleansing agent at the bedside of the diarrhea patient in the healthcare facility and provision of a soapy water bottle (water and detergent powder). Within 24 h of intervention delivery, a three-hour structured observation of handwashing practices at stool/vomit- and food-related events (key events) was conducted in healthcare facilities of diarrhea patients and their attendants. Compared to the standard arm, there was significantly more handwashing with a cleansing agent at key events in the PICHA7 arm (40% vs. 15%) (odds ratio: 5.04; (95% confidence interval (CI): 2.01, 12.7)). These findings demonstrate that delivery of the PICHA7 WASH pictorial module and provision of a soapy water bottle to diarrhea patients and their attendants presents a promising approach to increase handwashing with a cleansing agent among this high-risk population in healthcare facilities in the eastern DRC.
Topics: Humans; Diarrhea; Hand Disinfection; Male; Adult; Hygiene; Democratic Republic of the Congo; Female; Pilot Projects; Health Facilities; Sanitation; Middle Aged; Young Adult; Adolescent; Cholera
PubMed: 38928906
DOI: 10.3390/ijerph21060659 -
Foods (Basel, Switzerland) Jun 2024The increase in vegetable consumption has underlined the importance of minimizing the risks associated with microbiological contamination of fresh produce. The critical...
The increase in vegetable consumption has underlined the importance of minimizing the risks associated with microbiological contamination of fresh produce. The critical stage of the vegetable washing process has proven to be a key point for cross-contamination and the persistence of pathogens. In this context, the agri-food industry has widely adopted the use of disinfectants to reduce the bacterial load in the wash water. Therefore, we conducted laboratory-scale experiments in order to demonstrate the antimicrobial activity of disinfectants used in the wash tank of agro-food industries. Different wash water matrices of shredded lettuce, shredded cabbage, diced onion, and baby spinach were treated with sodium hypochlorite (NaClO), chlorine dioxide (ClO), and per-oxyacetic acid (PAA) at recommended concentrations. To simulate the presence of pathogenic bacteria, a cocktail of O157:H7 was inoculated into the process water samples (PWW) to determine whether concentrations of disinfectants inhibit the pathogen or bring it to a viable non-culturable state (VBNC). Hereby, we used quantitative qPCR combined with different photo-reactive dyes such as ethidium monoazide (EMA) and propidium monoazide (PMA). The results indicated that concentrations superior to 20 ppm NaClO inhibit the pathogen O157:H7 artificially inoculated in the process water. Concentrations between 10-20 ppm ClO fail to induce the pathogen to the VBNC state. At concentrations of 80 ppm PAA, levels of culturable bacteria and VBNC of O157:H7 were detected in all PWWs regardless of the matrix. Subsequently, this indicates that the recommended concentrations of ClO and PAA for use in the fresh produce industry wash tank do not inhibit the levels of O157:H7 present in the wash water.
PubMed: 38928858
DOI: 10.3390/foods13121915 -
Foods (Basel, Switzerland) Jun 2024Periodontal disease is an inflammatory disease caused by infection with periodontopathogenic bacteria. Oral care is essential to prevent and control periodontal disease,...
Periodontal disease is an inflammatory disease caused by infection with periodontopathogenic bacteria. Oral care is essential to prevent and control periodontal disease, which affects oral and systemic health. However, many oral hygiene products currently on the market were developed as disinfectants, and their intense irritation makes their use difficult for young children and older people. This study investigated the antibacterial effects of prunin laurate (Pru-C12) and its analogs on periodontopathogenic bacteria, (). Pru-C12 and its analogs inhibited in vitro bacterial growth at more than 10 μM and biofilm formation at 50 µM. Among its analogs, only Pru-C12 showed no cytotoxicity at 100 µM. Three of the most potent inhibitors also inhibited the formation of biofilms. Furthermore, Pru-C12 inhibited alveolar bone resorption in a mouse experimental periodontitis model by infection. These findings may be helpful in the development of oral hygiene products for the prevention and control of periodontal disease and related disorders.
PubMed: 38928857
DOI: 10.3390/foods13121917 -
Foods (Basel, Switzerland) Jun 2024Antimicrobial tolerance is a significant concern in the food industry, as it poses risks to food safety and public health. To overcome this challenge, synergistic...
Antimicrobial tolerance is a significant concern in the food industry, as it poses risks to food safety and public health. To overcome this challenge, synergistic combinations of antimicrobials have emerged as a potential solution. In this study, the combinations of two essential oil constituents (EOCs), namely carvacrol (CAR) and eugenol (EUG), with the quaternary ammonium compounds (QACs) benzalkonium chloride (BAC) and didecyldimethylammonium chloride (DDAC) were evaluated for their antimicrobial effects against and , two common foodborne bacteria. The checkerboard assay was employed to determine the fractional inhibitory concentration index (FICI) and the fractional bactericidal concentration index (FBCI), indicating the presence of bactericidal, but not bacteriostatic, synergy in all QAC-EOC combinations. Bactericidal synergism was clearly supported by Bliss independence analysis. The bactericidal activity of the promising synergistic combinations was further validated by time-kill curves, achieving a >4-log reduction of initial bacterial load, which is significant compared to typical industry standards. The combinations containing DDAC showed the highest efficiency, resulting in the eradication of bacterial population in less than 2-4 h. These findings emphasize the importance of considering both bacteriostatic and bactericidal effects when evaluating antimicrobial combinations and the potential of EOC-QAC combinations for sanitization and disinfection in the food industry.
PubMed: 38928773
DOI: 10.3390/foods13121831 -
International Journal of Molecular... Jun 2024An accumulation of reactive oxygen species (ROS) in cardiomyocytes can induce pro-arrhythmogenic late Na currents by removing the inactivation of voltage-gated Na...
An accumulation of reactive oxygen species (ROS) in cardiomyocytes can induce pro-arrhythmogenic late Na currents by removing the inactivation of voltage-gated Na channels including the tetrodotoxin (TTX)-resistant cardiac α-subunit Nav1.5 as well as TTX-sensitive α-subunits like Nav1.2 and Nav1.3. Here, we explored oxidant-induced late Na currents in mouse cardiomyocytes and human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) as well as in HEK 293 cells expressing Nav1.2, Nav1.3, or Nav1.5. Na currents in mouse cardiomyocytes and hiPSC-CMs treated with the oxidant chloramine T (ChT) developed a moderate reduction in peak current amplitudes accompanied by large late Na currents. While ChT induced a strong reduction in peak current amplitudes but only small persistent currents on Nav1.5, both Nav1.2 and Nav1.3 produced increased peak current amplitudes and large persistent currents following oxidation. TTX (300 nM) blocked ChT-induced late Na currents significantly stronger as compared to peak Na currents in both mouse cardiomyocytes and hiPSC-CMs. Similar differences between Nav1.2, Nav1.3, and Nav1.5 regarding ROS sensitivity were also evident when oxidation was induced with UVA-light (380 nm) or the cysteine-selective oxidant nitroxyl (HNO). To conclude, our data on TTX-sensitive Na channels expressed in cardiomyocytes may be relevant for the generation of late Na currents following oxidative stress.
Topics: Myocytes, Cardiac; Humans; Animals; Tetrodotoxin; Mice; Induced Pluripotent Stem Cells; HEK293 Cells; Oxidation-Reduction; Chloramines; Reactive Oxygen Species; NAV1.5 Voltage-Gated Sodium Channel; Sodium; Action Potentials; Tosyl Compounds
PubMed: 38928302
DOI: 10.3390/ijms25126596 -
International Journal of Molecular... Jun 2024Paraformaldehyde (PFA) fixation is the preferred method for preserving tissue architecture for anatomical and pathological observations. Meanwhile, PFA reacts with the...
Paraformaldehyde (PFA) fixation is the preferred method for preserving tissue architecture for anatomical and pathological observations. Meanwhile, PFA reacts with the amine groups of biomolecules to form chemical cross-linking, which preserves RNA within the tissue. This has great prospects for RNA sequencing to characterize the molecular underpinnings after anatomical and pathological observations. However, RNA is inaccessible due to cross-linked adducts forming between RNA and other biomolecules in prolonged PFA-fixed tissue. It is also difficult to perform reverse transcription and PCR, resulting in low sequencing sensitivity and reduced reproducibility. Here, we developed a method to perform RNA sequencing in PFA-fixed tissue, which is easy to use, cost-effective, and allows efficient sample multiplexing. We employ cross-link reversal to recover RNA and library construction using random primers without artificial fragmentation. The yield and quality of recovered RNA significantly increased through our method, and sequencing quality metrics and detected genes did not show any major differences compared with matched fresh samples. Moreover, we applied our method for gene expression analysis in different regions of the mouse brain and identified unique gene expression profiles with varied functional implications. We also find significant dysregulation of genes involved in Alzheimer's disease (AD) pathogenesis within the medial septum (MS)/vertical diagonal band of Broca (VDB) of the 5×FAD mouse brain. Our method can thus increase the performance of high-throughput RNA sequencing with PFA-fixed samples and allows longitudinal studies of small tissue regions isolated by their in situ context.
Topics: Formaldehyde; Animals; Mice; Brain; Tissue Fixation; Sequence Analysis, RNA; Alzheimer Disease; Polymers; Gene Expression Profiling; High-Throughput Nucleotide Sequencing; RNA
PubMed: 38928210
DOI: 10.3390/ijms25126504