Effect of Gamma Irradiation on Pathogenic Staphylococcus aureus in Packaged Ready-to-Eat Salads Treated with Biological Extracts.

Providing pathogen-free ready-to-eat (RTE) salads is critical for all consumers, especially individuals with weakened immunity. In this study, the efficacy of γ-irradiation on Staphylococcus aureus (S. aureus) in freshly packaged salads (4.24 log CFU/g) treated with essential oil (EO) and myrtle juice during 10 days of storage and their impact on organoleptic properties were investigated. EO was extracted by hydrodistillation and the chemical composition was analyzed by gas chromatography with Flame Ionization Detector (GC/FID) and gas chromatography/mass spectrometry (GC/MS). Myrtle juice was prepared from fresh fruits. The cytotoxic effects of Thymus capitatus (T. capitatus) EO against a normal human umbilical vein endothelial cell (HUVEC) were assessed. GC/FID and GC-MS analysis of the thyme EO revealed the presence of 13 compounds, including carvacrol (79.55%) and p-cymene (7.93%) as major components. The EO was found to be noncytotoxic, with concentrations lower than 0.16 µL/mL. A reduction of more than 3 log CFU/g and a total inactivation of S. aureus were achieved with the combination of gamma irradiation at 0.5 kGy with myrtle juice at 6 µL/mL and EO at 0.08 µL/mL, respectively. The treatment of fresh RTE salads with thyme and myrtle juice was evaluated as acceptable by the sensory panel. The combined effect showed a synergistic potential on the inactivation of S. aureus.

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.

Occurrence of Staphylococcus spp. and investigation of fecal and animal viral contaminations in livestock, river water, and sewage from Tunisia and Romania.

The objective of this study was to determine the occurrence of Staphylococcus spp., Escherichia coli, somatic coliphages, F-specific RNA bacteriophages, hepatitis E virus (HEV), and bovine enterovirus (BEV) in fecal and water samples. The occurrence of Staphylococcus spp. was investigated in treated wastewater samples collected from slaughterhouse of Tunisia. Results showed that Staphylococcus spp. were detected in the totality of collected samples with an average mean of 5.44 Log10 (CFU/100 ml). Regarding fecal indicator bacteria, E. coli was detected in the totality of water samples and was more abundant in Tunisian samples than in samples collected from Romania (P < 0.05). Concerning somatic coliphages and F-specific RNA bacteriophages used as viral indicators, they were detected in all raw and treated wastewaters. Bovine enterovirus (BEV) was detected in 20.1% and 28% of bovine stool samples collected from Tunisia and Romania, respectively. BEV was also detected in 60% of porcine stool samples from Romania. BEV was absent in all treated sewage samples. HEV was detected in raw sewage and bovine fecal sample from Romania with low occurrence and none sample from Tunisia was positive. This study may give us an insight into the monitoring of water quality in Tunisia and Romania.

Comparison of the Performance of Different Microbial Source Tracking Markers among European and North African Regions.

Microbial source tracking (MST) has been extensively used to detect the sources of fecal pollution in water. The inclusion of MST in water management strategies improves the ecological status of the ecosystem and human and animal health under interdisciplinary analysis in all aspects of health care for humans, animals, and the environment (One Health approach). In this study, the performance of MST markers targeting host-specific Bacteroidales (HF183 and Rum-2-Bac) and species (HMBif and CWBif) were evaluated in raw sewage collected from human, ruminant, swine, and poultry sources in Tunisia, Cyprus, Ireland, and Spain. In addition, the ratio between somatic coliphages and bacteriophages infecting GA17 (SOMCPH/GA17PH) was measured in Tunisia and Spain. The obtained results showed variability of the bacterial markers between the four countries, suggesting that their usefulness could be affected by several conditions (dietary habits, agricultural practices, and climatic conditions) that differ between countries. The Rum-2-Bac marker stood out as a valid MST tool, particularly in Ireland, whereas CWBif was the best option in Tunisia, Spain, and Cyprus. The human-specific HMBif marker showed good sensitivity and specificity in Tunisia, Spain, and Ireland, whereas HF183 showed a low specificity. However, HF183 was suggested as a good human marker in Ireland and Cyprus because of its higher concentration than HMBif. Regarding viral markers, the ratio of SOMCPH/GA17PH showed a clear discrimination between human and nonhuman samples. The combined use of molecular bacterial markers and the ratio of SOMCPH/GA17PH may improve the success of MST.

Total coliphages removal by activated sludge process and their morphological diversity by transmission electron microscopy.

This study was conducted to isolate phages in treated sewage collected from wastewater treatment plant, and explore their morphological diversity by transmission electron microscopy (TEM). Fates of total bacteriophages and their reduction by biological treatment were also assayed. Phages were isolated using the plaque assay then negatively stained and observed by electron microscope. Electron micrographs showed different types of phages with different shapes and sizes. The majority of viruses found in treated sewage ranged from 30 to 100 nm in capsid diameter. Many of them were tailed, belonging to Siphoviridae, Myoviridae and Podoviridae families. Non-tailed phage particles were also found at a low rate, presumably belonging to Leviviridae or Microviridae families. This study shows the diversity and the abundance of bacteriophages in wastewater after biological treatment. Their persistence in wastewater reused in agriculture should raise concerns about their potential role in controlling bacterial populations in the environment. They should be also included in water treatment quality controlling guidelines as fecal and viral indicators.

A comparison of two extraction methods for the detection of Enteroviruses in raw sludge.

The aim of this study was to compare two viral extraction methods for the detection of naturally occurring Enteroviruses in raw sludge. The first method (M1) is based on an ultracentrifugation step. In the second one (M2), viral RNA was extracted directly after viral elution from suspended solids. Genomes of enteroviruses were quantified by a quantitative real time PCR (qRT-PCR) in sludge samples. Somatic coliphages and F-specific RNA phages, considered as viral indicators of enteric viruses in sludge, were enumerated by the double layer agar technique. Results showed that direct assay of RNA extraction yielded higher genomic copies of enteric viruses (with an average of 5.07Log10 genomic copies/100 mL). After the ultracentrifugation assay in the second method, genomic copies number decreases (with an average of 4.39Log10 genomic copies/100 mL). This can be explained by an eventual concentration of inhibitors existing in sludge samples. Phages enumeration results showed their presence in all sludge samples with an average of (5.69Log10 PFU/100 mL) for somatic coliphages and (4Log10 PFU/100 mL) for F-specific RNA phages. This emphasizes the use of somatic coliphages as viral indicators for enteroviruses in environmental samples and especially in raw sludge samples in wastewater treatment plants prior to agricultural use.