Red cabbage extract immobilized in bacterial cellulose film as an eco-friendly sensor to monitor microbial contamination and gamma irradiation of stored cucumbers.

The aim of the present study is to develop a pH-sensing biopolymer film based on the immobilization of red cabbage extract (RCE) within bacterial cellulose (BC) to detect contamination and gamma radiation exposure in cucumbers. The results obtained show a sensitivity to pH changes for RCE in its aqueous form and that incorporated within BC films (RCE-BC), both showed color change correlated to bacterial growth (R2 = 0.91), this was supported with increase in pH values from 2 to 12 (R2 = 0.98). RCE and RCE-BC exposure to gamma radiation (0, 2.5, 5, 10, 15, 20, 25 kGy) resulted in gradual decrease in color that was more evident in RCE aqueous samples. To sense bacterial contamination of cucumbers, the total count was followed at 0, 5, 10 and 15 days in cold storage conditions and was found to reach 9.13 and 5.47 log cfu/mL for non-irradiated and 2 kGy irradiated samples, respectively. The main isolates detected throughout this storage period were identified as Pseudomonas fluorescens, Erwinia sp. Pantoea agglomerans using matrix assisted laser desorption ionization-time of flight-ms (MALDI-TOF-MS). Bacterial growth in stored irradiated cucumbers was detected by color change within 5 and 10 days of storage, after which there was no evident change. This is very useful since contamination within the early days of storage cannot be sensed with the naked eye. This study is the first to highlight utilizing RCE and RCE-BC as eco-friendly pH-sensing indicator films for intelligent food packaging to detect both food contamination and gamma preservation for refrigerator stored cucumbers.

Dramatic loss of microbial viability in bentonite exposed to heat and gamma radiation: implications for deep geological repository.

Bentonite is an integral part of the engineered barrier system (EBS) in deep geological repositories (DGR) for nuclear waste, but its indigenous microorganisms may jeopardize long-term EBS integrity. To predict microbial activity in DGRs, it is essential to understand microbial reactions to the early hot phase of DGR evolution. Two bentonites (BCV and MX-80) with varied bentonite/water ratios and saturation levels (compacted to 1600 kg.m- 3 dry density/powder/suspension), were subjected to heat (90-150 °C) and irradiation (0.4 Gy.h- 1) in the long-term experiments (up to 18 months). Molecular-genetic, microscopic, and cultivation-based techniques assessed microbial survivability. Exposure to 90 °C and 150 °C notably diminished microbial viability, irrespective of bentonite form, with negligible impacts from irradiation or sample type compared to temperature. Bentonite powder samples exhibited microbial recovery after 90 °C heating for up to 6 months but not 12 months in most cases; exposure to 150 °C had an even stronger effect. Further long-term experiments at additional temperatures combined with the mathematical prediction of temperature evolution in DGR are recommended to validate the possible evolution and spatial distribution of microbially depleted zones in bentonite buffer around the waste canisters and refine predictions of microbial effects over time in the DGR.

Effectiveness of UV-C radiation in inactivation of microorganisms on materials with different surface structures.

INTRODUCTION AND OBJECTIVE: Ultraviolet light in the UV-C band is known as germicidal radiation and was widely used for both sterilization of the equipment and creation of a sterile environment. The aim of the study is to assess the effectiveness of inactivation of microorganisms deposited on surfaces with various textures by UV-C radiation disinfection devices. MATERIAL AND METHODS: Five microorganisms (3 bacteria, virus, and fungus) deposited on metal, plastic, and glass surfaces with smooth and rough textures were irradiated with UV-C light emitted by low-pressure mercury lamp and ultraviolet emitting diodes (LEDs), from a distance of 0.5 m, 1 m, and 1.5 m to check their survivability after 20-minute exposure. RESULTS AND CONCLUSIONS: Both tested UV-C sources were effective in inactivation of microorganisms; however, LED emitter was more efficient in this respect than the mercury lamp. The survival rate of microorganisms depended on the UV-C dose, conditioned by the distance from UV-C source being the highest at 0.5 m and the lowest at 1.5 m. For the tested microorganisms, the highest survival rate after UV-C irradiation was usually visible on glass and plastic surfaces. This observation should be considered in all environments where the type of material (from which the elements of technical equipment are manufactured and may be contaminated by specific activities) is important for maintaining the proper level of hygiene and avoiding the unwanted and uncontrolled spread of microbiological pollution.

Enhanced antimicrobial efficacy and energy efficiency of low irradiance 405-nm light for bacterial decontamination.

Due to its increased safety over ultraviolet light, there is interest in the development of antimicrobial violet-blue light technologies for infection control applications. To ensure compatibility with exposed materials and tissue, the light irradiances and dose regimes used must be suitable for the target application. This study investigates the antimicrobial dose responses and germicidal efficiency of 405 nm violet-blue light when applied at a range of irradiance levels, for inactivation of surface-seeded and suspended bacteria. Bacteria were seeded onto agar surfaces (101-108 CFUplate-1) or suspended in PBS (103-109 CFUmL-1) and exposed to increasing doses of 405-nm light (≤ 288 Jcm-2) using various irradiances (0.5-150 mWcm-2), with susceptibility at equivalent light doses compared. Bacterial reductions ≥ 96% were demonstrated in all cases for lower irradiance (≤ 5 mWcm-2) exposures. Comparisons indicated, on a per unit dose basis, that significantly lower doses were required for significant reductions of all species when exposed at lower irradiances: 3-30 Jcm-2/0.5 mWcm-2 compared to 9-75 Jcm-2/50 mWcm-2 for low cell density (102 CFUplate-1) surface exposures and 22.5 Jcm-2/5 mWcm-2 compared to 67.5 Jcm-2/150 mWcm-2 for low density (103 CFUmL-1) liquid exposures (P ≤ 0.05). Similar patterns were observed at higher densities, excluding S. aureus exposed at 109 CFUmL-1, suggesting bacterial density at predictable levels has minimal influence on decontamination efficacy. This study provides fundamental evidence of the greater energy efficacy of 405-nm light for inactivation of clinically-significant pathogens when lower irradiances are employed, further supporting its relevance for practical decontamination applications.

Optimized Ultraviolet-C Processing Inactivates Pathogenic and Spoilage-Associated Bacteria while Preserving Bioactive Proteins, Vitamins, and Lipids in Human Milk.

Holder pasteurization (HoP) enhances donor human milk microbiological safety but damages many bioactive milk proteins. Though ultraviolet-C irradiation (UV-C) can enhance safety while better preserving some milk proteins, it has not been optimized for dose or effect on a larger array of bioactive proteins. We determined the minimal UV-C parameters that provide >5-log reductions of relevant bacteria in human milk and how these treatments affect an array of bioactive proteins, vitamin E, and lipid oxidation. Treatment at 6000 and 12 000 J/L of UV-C resulted in >5-log reductions of all vegetative bacteria and bacterial spores, respectively. Both dosages improved retention of immunoglobulin A (IgA), IgG, IgM, lactoferrin, cathepsin D, and elastase and activities of bile-salt-stimulated lipase and lysozyme compared with HoP. These UV-C doses caused minor reductions in α-tocopherol but not γ-tocopherol and no increases in lipid oxidation products. UV-C treatment is a promising approach for donor human milk processing.

Light over mechanics: microbial community structure and activity in simulated migrating bedforms are controlled by oscillating light rather than by mechanical forces.

Sandy sediments of lowland streams are transported as migrating ripples. Benthic microorganisms colonizing sandy grains are exposed to frequent moving-resting cycles and are believed to be shaped by two dominant environmental factors: mechanical stress during the moving phase causing biofilm abrasion, and alternating light-dark cycles during the resting phase. Our study consisted of two laboratory experiments and aimed to decipher which environmental factor causes the previously observed hampered sediment-associated microbial activity and altered community structure during ripple migration. The first experiment tested the effect of three different migration velocities under comparable light conditions. The second experiment compared migrating and stationary sediments under either constant light exposure or light oscillation. We hypothesized that microbial activity and community structure would be more strongly affected by (1) higher compared to lower migration velocities, and by (2) light oscillation compared to mechanical stress. Combining the results from both experiments, we observed lower microbial activity and an altered community structure in sediments exposed to light oscillation, whereas migration velocity had less impact on community activity and structure. Our findings indicate that light oscillation is the predominating environmental factor acting during ripple migration, resulting in an increased vulnerability of light-dependent photoautotrophs and a possible shift toward heterotrophy.

Acute Impacts of Ionizing Radiation Exposure on the Gastrointestinal Tract and Gut Microbiome in Mice.

Radiation therapy for abdominopelvic malignancies often results in damage to the gastrointestinal tract (GIT) and permanent changes in bowel function. An overlooked component of the pathophysiology of radiation-induced bowel injury is the role of the gut microbiome. The goal of this research was to identify the impacts of acute radiation exposure on the GIT and gut microbiome. C57BL/6 mice exposed to whole-body X-rays (0.1-3 Gy) were assessed for histological and microbiome changes 48 h post-radiation exposure. Within the ileum, a dose of 3 Gy significantly decreased crypt depth as well as the number of goblet cells, but increased overall goblet cell size. Overall, radiation altered the microbial distribution within each of the main phyla in a dose- and tissue-dependent manner. Within the Firmicutes phylum, high dose irradiation resulted in significant alterations in bacteria from the class Bacilli within the small bowels, and from the class Clostridia in the large bowels. The 3 Gy radiation also significantly increased the abundance of bacterial families from the Bacteroidetes phylum in the colon and feces. Overall, we identified various alterations in microbiome composition following acute radiation exposure, which could potentially lead to novel biomarkers for tracking patient toxicities or could be used as targets for mitigation strategies against radiation damage.

Using UV/peracetic acid as pretreatment for subsequent bio-treatment of antibiotic-containing wastewater treatment: Mitigating microbial inhibition and antibiotic resistance genes proliferation.

UV/peracetic acid (PAA) treatment presents a promising approach for antibiotic removal, but its effects on microbial community and proliferation of antibiotic resistance genes (ARGs) during the subsequent bio-treatment remain unclear. Thus, we evaluated the effects of the UV/PAA on tetracycline (TTC) degradation, followed by introduction of the treated wastewater into the bio-treatment system to monitor changes in ARG expression and biodegradability. Results demonstrated effective TTC elimination by the UV/PAA system, with carbon-centered radicals playing a significant role. Crucially, the UV/PAA system not only eliminated antibacterial activity but also inhibited potential ARG host growth, thereby minimizing the emergence and dissemination of ARGs during subsequent bio-treatment. Additionally, the UV/PAA system efficiently removed multi-antibiotic resistant bacteria and ARGs from the bio-treatment effluent, preventing ARGs from being released into the environment. Hence, we propose a multi-barrier strategy for treating antibiotic-containing wastewater, integrating UV/PAA pre-treatment and post-disinfection with bio-treatment. The inhibition of ARGs transmission by the integrated system was verified through actual soil testing, confirming its effectiveness in preventing ARGs dissemination in the surrounding natural ecosystem. Overall, the UV/PAA treatment system offers a promising solution for tackling ARGs challenges by controlling ARGs proliferation at the source and minimizing their release at the end of the treatment process.

Efficacy of 233 nm LED far UV-C-radiation against clinically relevant bacterial strains in the phase 2/ step 2 in vitro test on basis of EN 14561 and on an epidermis cell model.

INTRODUCTION: Healthcare-acquired infections and overuse of antibiotics are a common problem. Rising emergence of antibiotic and antiseptic resistances requires new methods of microbial decontamination or decolonization as the use of far-UV-C radiation. METHODS: The microbicidal efficacy of UV-C radiation (222 nm, 233 nm, 254 nm) was determined in a quantitative carrier test and on 3D-epidermis models against Staphylococcus (S.) aureus, S.epidermidis, S.haemolyticus, S.lugdunensis, Klebsiella pneumoniae, and Pseudomonas aeruginosa. To mimic realistic conditions, sodium chloride solution, mucin, albumin, artificial saliva, artificial wound exudate and artificial sweat were used. RESULTS: In sodium chloride solution, irradiation with a dose of 40 mJ/cm2 (233 nm) was sufficient to achieve 5 lg reduction independent of bacteria genus or species. In artificial sweat, albumin and artificial wound exudate, a reduction >3 lg was reached for most of the bacteria. Mucin and artificial saliva decreased the reduction to <2 lg. On 3D epidermis models, reduction was lower than in the carrier test. CONCLUSION: UV-C radiation at 233 nm was proven to be efficient in bacteria inactivation independent of genus or species thus being a promising candidate for clinical use in the presence of humans and on skin/mucosa.

Photoinactivation of microorganisms using bacteriochlorins as photosensitizers.

In recent years, some microorganisms have shown resistance to conventional treatments. Considering this increase in resistant pathogens, treatment alternatives are needed to promote greater treatment efficiency. In this sense, antimicrobial photodynamic therapy (aPDT) has been an alternative treatment. This technique uses a photosensitizer that is activated by light with a specific wavelength producing reactive species, leading to the death of pathogenic microorganisms. In this study, bacteriochlorophyll derivatives such as bacteriochlorin metoxi (Bchl-M) and bacteriochlorin trizma (Bchl-T) obtained from purple bacterium (Rhodopseudomonas faecalis), were evaluated as photosensitizers in the aPDT. Photodynamic inactivation (PDI) of the microorganisms Staphylococcus aureus, Micrococcus luteus, Candida albicans and Pseudomonas aeruginosa was investigated with both bacteriochlorins (Bchl-M and Bchl-T) at different concentrations (1, 15 and 30 µM for S. aureus; 1, 15, 30, 45, 60 and 75 µM for M. luteus; 30, 60, 90, 105, 120 and 150 µM for C. albicans; and 200 µM for P. aeruginosa) and different doses of light (20 and 30 J/cm2 for S. aureus and M. luteus; 30 and 45 J/cm2 for C. albicans; and 45 J/cm2 for P. aeruginosa) to inactivate them. Both photosensitizers showed good activation against S. aureus and for M. luteus, we observed the inactivation of these microorganisms at approximately 3 log, showing to be a good photosensitizers for these microorganisms.

Influence of MHz-order acoustic waves on bacterial suspensions.

The development of alternative techniques to efficiently inactivate bacterial suspensions is crucial to prevent transmission of waterborne illness, particularly when commonly used techniques such as heating, filtration, chlorination, or ultraviolet treatment are not practical or feasible. We examine the effect of MHz-order acoustic wave irradiation in the form of surface acoustic waves (SAWs) on Gram-positive (Escherichia coli) and Gram-negative (Brevibacillus borstelensis and Staphylococcus aureus) bacteria suspended in water droplets. A significant increase in the relative bacterial load reduction of colony-forming units (up to 74%) can be achieved by either increasing (1) the excitation power, or, (2) the acoustic treatment duration, which we attributed to the effect of the acoustic radiation force exerted on the bacteria. Consequently, by increasing the maximum pressure amplitude via a hybrid modulation scheme involving a combination of amplitude and pulse-width modulation, we observe that the bacterial inactivation efficiency can be further increased by approximately 14%. By combining this scalable acoustic-based bacterial inactivation platform with plasma-activated water, a 100% reduction in E. coli is observed in less than 10 mins, therefore demonstrating the potential of the synergistic effects of MHz-order acoustic irradiation and plasma-activated water as an efficient strategy for water decontamination.

Ultraviolet-C light at 222 nm has a high disinfecting spectrum in environments contaminated by infectious pathogens, including SARS-CoV-2.

Ultraviolet light (UV) acts as a powerful disinfectant and can prevent contamination of personal hygiene from various contaminated environments. The 222-nm wavelength of UV-C has a highly effective sterilization activity and is safer than 275-nm UV-C. We investigated the irradiation efficacy of 222-nm UV-C against contaminating bacteria and viruses in liquid and fabric environments. We conducted colony-forming unit assays to determine the number of viable cells and a 50% tissue culture infectious dose assay to evaluate the virus titration. A minimum dose of 27 mJ/cm2 of 222-nm UV-C was required for >95% germicidal activity for gram-negative and -positive bacteria. A 25.1 mJ/cm2 dose could ensure >95% virucidal activity against low-pathogenic avian influenza virus and severe acute respiratory syndrome coronavirus (SARS-CoV-2). In addition, this energy dose of 222-nm UV-C effectively inactivated SARS-CoV-2 variants, Delta and Omicron. These results provide valuable information on the disinfection efficiency of 222-nm UV-C in bacterial and virus-contaminated environments and can also develop into a powerful tool for individual hygiene.

Phototrophy by antenna-containing rhodopsin pumps in aquatic environments.

Energy transfer from light-harvesting ketocarotenoids to the light-driven proton pump xanthorhodopsins has been previously demonstrated in two unique cases: an extreme halophilic bacterium1 and a terrestrial cyanobacterium2. Attempts to find carotenoids that bind and transfer energy to abundant rhodopsin proton pumps3 from marine photoheterotrophs have thus far failed4-6. Here we detected light energy transfer from the widespread hydroxylated carotenoids zeaxanthin and lutein to the retinal moiety of xanthorhodopsins and proteorhodopsins using functional metagenomics combined with chromophore extraction from the environment. The light-harvesting carotenoids transfer up to 42% of the harvested energy in the violet- or blue-light range to the green-light absorbing retinal chromophore. Our data suggest that these antennas may have a substantial effect on rhodopsin phototrophy in the world's lakes, seas and oceans. However, the functional implications of our findings are yet to be discovered.

An excimer lamp to provide far-ultraviolet C irradiation for dining-table disinfection.

Dining tables may present a risk to diners by transmitting bacteria and/or viruses. Currently, there is a lack of an environmental-friendly and convenient means to protect diners when they are sitting together. This investigation constructed far-UVC excimer lamps to disinfect dining-table surfaces. The lamps were mounted at different heights and orientations, and the irradiance on table surfaces was measured. The irradiation doses to obtain different inactivation efficiencies for Escherichia coli (E. coli) were provided. In addition, numerical modeling was conducted for irradiance and the resulting inactivation efficiency. The surface-to-surface (S2S) model was validated with the measured irradiance. The germicidal performance of far-UVC irradiation, the far-UVC doses to which diners were exposed, and the risk of exposure to the generated ozone were evaluated. The results revealed that an irradiation dose of 12.8 mJ/cm2 can disinfect 99.9% of E. coli on surfaces. By varying the lamp irradiance output, the number and positions of the lamps, the far-UVC irradiation can achieve a 3-log reduction for a dining duration of 5 min. Besides, the far-UVC lamp has a low damage risk to diners when achieving an effective inactivation rate. Moreover, there is virtually no ozone exposure risk in a mechanically ventilated dining hall.

A novel exposure mode based on UVA-LEDs for bacterial inactivation.

As an emerging UV source, ultraviolet light-emitting diodes (UV-LEDs) are increasingly being used for disinfection purposes. UVA-LEDs have a higher output power, lower cost, and stronger penetration and cause less harm than UVC-LEDs. In this study, a novel exposure mode based on UVA was proposed and well demonstrated by various experiments using S. aureus as an indicator. Compared with single-dose exposure, fractionated exposure with a 15 min interval between treatments resulted in increased S. aureus inactivation. A longer interval or lower first irradiation dose was unfavorable for inactivation. Fractionated exposure changed the inactivation rate constant and eliminated the shoulder in the fluence-response curves. This resulted in changing the sensitivity of bacteria to UVA and improving bacterial inactivation. Moreover, the fractioned exposure mode has universality for various bacteria (including gram-positive and gram-negative bacteria). S. aureus was not reactivated by photoreactivation or dark repair after UVA treatment. As expected, the cells were damaged more seriously after fractionated exposure, further suggesting the advantages of this new exposure mode. In addition, the mechanism by which bacteria were inactivated after fractionated exposure was investigated, and it was found that •OH played an important role. A longer interval between treatments showed an adverse effect on inactivation, mainly due to the reduction of •OH and recovery of intracellular GSH. In summary, the current work provides novel ideas for the application of UVA-LEDs, which will give more choices for disinfection treatment.

UV Inactivation of Common Pathogens and Surrogates Under 222 nm Irradiation from KrCl* Excimer Lamps.

Germicidal ultraviolet (UV) devices have been widely used for pathogen disinfection in water, air, and on food and surfaces. Emerging UV technologies, like the krypton chloride (KrCl*) excimer emitting at 222 nm, are rapidly gaining popularity due to their minimal adverse effects on skin and eyes compared with conventional UV lamps emitting at 254 nm, opening opportunities for UV disinfection in occupied public spaces. In this study, inactivation of seven bacteria and five viruses, including waterborne, foodborne and respiratory pathogens, was determined in a thin-film aqueous solution using a filtered KrCl* excimer emitting primarily at 222 nm. Our results show that the KrCl* excimer can effectively inactivate all tested bacteria and viruses, with most microorganisms achieving more than 4-log (99.99%) reduction with a UV dose of 10 mJ cm-2 . Compared with conventional UV lamps, the KrCl* excimer lamp exhibited better disinfection performance for viruses but was slightly less effective for bacteria. The relationships between UV sensitivities at 222 and 254 nm for bacteria and viruses were evaluated using regression analysis, resulting in factors that could be used to estimate the KrCl* excimer disinfection performance from well-documented UV kinetics using conventional 254 nm UV lamps. This study provides fundamental information for pathogen disinfection when employing KrCl* excimers.

Inactivation of antibiotic resistant bacteria from stormwater runoff using UVA/LED and its potential risks.

Recently, increasing attention has been paid to antibiotic resistance in stormwater runoff. However, there is no available literature about the control of antibiotic resistant bacteria (ARB) through 365 nm ultraviolet light-emitting diode (UVA/LED). In this study, batch experiments were conducted to investigate ARB inactivation kinetics, effects of light intensity and water matrix (including suspended solid (SS) concentration, initial pH and bacteria concentration), and potential transmission risks after UVA/LED irradiation. Results showed that ARB inactivation efficiencies reached 6.31 log reduction at 8 mW/cm2 (86 J/cm2) of UVA/LED for 180 min. ARB inactivation efficiencies increased with the increase of light intensity, and showed a linear relationship. ARB inactivation decreased with increasing SS levels, and the largest inactivation efficiencies was 3.56 log reduction at 50 mg/L of SS. Initial pH had slight effect on ARB inactivation through UVA/LED irradiation. A low initial bacteria concentration (105 CFU/mL) was not necessarily associated with good ARB inactivation (3.59 log reduction). After UVA/LED irradiation, ARB was hardly detected during 12 hr of dark repair, and the transfer frequency of kanamycin resistance gene was increased to 5.43 × 10-4. These suggested that the application of UVA/LED to inactivate ARB in stormwater runoff was feasible and desirable in this study.

UV C Light from a Light-Emitting Diode at 275 Nanometers Shortens Wound Healing Time in Bacterium- and Fungus-Infected Skin in Mice.

Due to the changes in pathogenic species and the absence of research on topical skin antibiotics, the therapy of skin and soft tissue infections (SSTIs) is facing more and more severe challenges. It is particularly urgent to look for alternative therapies without induction of drug resistance. UV C (UVC) light within the range of 200 to 280 nm is one of the most common techniques used to kill and/or inactivate pathogenic microorganisms. However, the traditional most commonly used wavelength of 254 nm irradiated from a low-pressure mercury lamp is hazardous to human health, being both carcinogenic and damaging to eye tissues, which limits its applications in vivo. This research aimed to investigate the antimicrobial properties and influence of 275-nm UVC light from a light-emitting diode (UVC-LED light) on wound healing time. Five bacteria, three fungi, and scalded-mouse models combined with SSTIs were used to evaluate the antimicrobial effect in vitro and in vivo. 275-nm UVC-LED light inactivated both bacteria and fungi with a very short irradiation time in vitro and induced neither DNA damage nor epidermal lesions in the mice's skin. Furthermore, in mouse models of SSTIs induced by either methicillin-resistant Staphylococcus aureus (MRSA) or Candida albicans, the 275-nm UVC-LED light showed significant antimicrobial effects and shortened the wound healing time compared with that in the no-irradiation group. UVC-LED light at 275 nm has the potential to be a new form of physical therapy for SSTIs. IMPORTANCE As a common clinical problem, the therapy of SSTIs is facing growing challenges due to an increase in the number of drug-resistant bacteria and fungi. UV C (UVC) light sterilization has been widely used in all aspects of daily life, but there are very few reports about in vivo therapy using UVC light. It is well known that prolonged exposure to UVC light increases the possibility of skin cancer. In addition, it is also very harmful for eyes. UV irradiation with 254-nm UVC light can cause corneal damage, like thinning of the corneal epithelial layer, superficial punctate keratitis, corneal erosion, etc. In this study, we focused on looking for a more accessible light source and safer UVC wavelength, and 275-nm UVC LED light was chosen. We investigated its applicability for SSTIs therapy with relative skin safety and expected that it could be used as a new physical therapy method for SSTIs.

Inactivation of biohazards in healthcare wastewater by E-Beam and Gamma irradiation: a comparative study.

The main objective of this study is to evaluate the effect of irradiation by Gamma rays and Electron Beam (E-Beam) on naturally occurring microorganisms shed in healthcare wastewater issued from multi-specialties hospital. We examined the susceptibility of naturally occurring total indicator bacteriophages towards Gamma rays and E-Beam irradiation to evaluate their appropriateness as viral indicators for healthcare wastewater quality control. Results showed that healthcare wastewater is a rich matrix containing bacteriophages surrogates of pathogenic waterborne viruses (4.5 Log10 PFU/100 mL for SOMCPH and 2.3 Log10 PFU/100 mL for FRNAPH), antibiotic resistant bacteria (Mean concentrations from 2.3 to 5.5 Log10 CFU/100 mL), molds and yeasts (2.7 Log10 CFU/100 mL), and spores of Clostridium perfringens (Mean concentration of 3.3 Log10 CFU/100 mL). After E-Beam irradiation, naturally occurring bacteria in healthcare wastewater showed lower resistance patterns (D10 values ranging between 0.21 ± 0.005 and 0.59 ± 0.005) compared to those obtained after Gamma irradiation (D10 values ranging between 0.25 ± 0.015 and 0.70 ± 0.0001). Spores of Clostridium perfringens were the most resistant assayed microbes either after E-Beam (D10 values of 3.74 ± 0.005) or Gamma irradiation (D10 values of 4.77 ± 0.025) of collected samples. According to inactivation patterns, a dose of 10 kGy was sufficient for a complete inactivation of spores. Bacteriophages isolated from healthcare wastewater showed the same resistance patterns as those previously obtained in urban treated sewage and were inactivated using higher doses than waterborne bacteria (D10 values of SOMCPH 1.46 ± 0.057; D10 values of FRNAPH 1.03 ± 0.057). Their resistance to irradiation treatment in such complex matrix corroborates their use to survey the viral quality of healthcare wastewater before their discharge in the urban sanitation network. D10 value analysis showed that bacteria and bacteriophages inactivation by E-Beam irradiation required lower doses than those required for their inactivation using Gamma rays. According to inactivation patterns, a dose of 7 kGy was sufficient for total inactivation of both pathogenic bacteria and viruses. Thus, E-Beam irradiation seems to be an efficient physical pre-treatment process for healthcare wastewater treatment prior to its discharge in urban sanitation system to ensure compliance with environmental standards and protect public health.