Inactivation of two oyster pathogens by photocatalysis and monitoring of changes in the microbiota of seawater: A case study on Ostreid herpes virus 1 µVar and Vibrio harveyi.

The pollution of seawater by both biotic (bacteria, viruses) and abiotic contaminants (biocides, pharmaceutical residues) frequently leads to economic losses in aquaculture activities mostly mortality events caused by microbial infection. Advanced Oxidation Processes (AOPs) such as heterogeneous photocatalysis allow the removal of all organic contaminants present in water and therefore could reduce production losses in land-based farms. Oysters in land-based farms such as hatcheries and nurseries suffer from a large number of mortality events, resulting in significant losses. If photocatalysis has been widely studied for the decontamination, its application for disinfection is still overlooked, especially on seawater for viruses. We therefore studied seawater disinfection using the photocatalysis (UV365/TiO2) method in the context of Pacific oyster mortality syndrome (POMS). POMS has been defined as a polymicrobial disease involving an initial viral infection with Ostreid Herpes Virus 1, accompanied by multiple bacterial infections. We investigated the impact of treatment on Vibrio harveyi, a unique opportunistic pathogenic bacterium, and on a complex microbial community reflecting a natural POMS event. Viral inactivation was monitored using experimental infections to determine whether viral particles were still infectious after. Changes in the total bacterial community in seawater were studied by comparing UV365/TiO2 treatment with UV365-irradiated seawater and untreated seawater. In the case of OsHV-1, a 2-h photocatalytic treatment prevents POMS disease and oyster mortality. The same treatment also inactivates 80% of viable Vibrio harveyi culture (c.a. 1.5 log). Since OsHV-1 and Vibrio harveyi are effectively inactivated without long-term destabilization of the total bacterial microbiota in the seawater, photocatalysis appears to be a relevant alternative for disinfecting seawater in land-based oyster beds.

Removal of pathogens by ultrafiltration from sea water.

Among water treatment processes, ultrafiltration is known to be efficient for the elimination of micro-organisms (bacteria and viruses). In this study, two pathogens were targeted, a bacterium, Vibrio aestuarianus and a virus, OsHV-1, with the objective to produce high quality water from seawater, in the case of shellfish productions. The retention of those microorganisms by ultrafiltration was evaluated at labscale. In the case of OsHV-1, the protection of oysters was validated by in vivo experiments using oysters spat and larvae, both stages being highly susceptible to the virus. The oysters raised using contaminated seawater which was then subsequently treated by ultrafiltration, had similar mortality to the negative controls. In the case of V. aestuarianus, ultrafiltration allowed a high retention of the bacteria in seawater with concentrations below the detection limits of the 3 analytical methods (flow cytometry, direct seeding and seeding after filtration to 0.22 µm). Thus, the quantity of V. aestuarianus was at least, 400 times inferior to the threshold known to induce mortalities in oysters. Industrial scale experiment on a several months period confirmed the conclusion obtained at lab scale on the Vibrio bacteria retention. Indeed, no bacteria from this genus, potentially harmful for oysters, was detected in permeate and this, whatever the quality of the seawater treated and the bacteria concentration upstream of the membrane. Moreover, the resistance of the process was confirmed with a stability of hydraulic performances over time for two water qualities and even facing an algal bloom. In terms of retention and resistance, ultrafiltration process was validated for the treatment of seawater towards the targeted pathogenic microorganisms, with the aim of biosecuring shellfish productions.

Optimization of Air Backwash Frequency during the Ultrafiltration of Seawater.

The main objective of this paper is to study the effect of new air backwash on dead-end ultrafiltration of seawater with a pilot at semi-industrial scale (20 m3/day). To control membrane fouling, two different backwashes were used to clean the membrane: classical backwash (CB) and new air backwash (AB) that consists of injecting air into the membrane module before a classical backwash. To evaluate the efficiency of AB and CB, a resistance in series model was used to calculate each resistance: membrane (Rm), reversible (Rrev) and irreversible (Rirr). The variation of the seawater quality was considered by integrating the turbidity variation versus time. The results indicate clearly that AB was more performant than CB and frequency of AB/CB cycles was important to control membrane fouling. In this study, frequencies of 1/5 and 1/3 appear more efficient than 1/7 and 1/9. In addition, the operation conditions (flux and time of filtration) had an important role in maintaining membrane performance-whatever the variation of the seawater quality.

Air Backwash Efficiency on Organic Fouling of UF Membranes Applied to Shellfish Hatchery Effluents.

Among all the techniques studied to overcome fouling generated in dead-end filtration, the injection of air during backwashes proved to be the most effective. Indeed, shear stress engendered by the two-phase flow enhanced particle removal on membrane surface. This work aims to study the injection of air to drain the membranes before backwash. Firstly, the efficiency of this backwash procedure was evaluated during the ultrafiltration of seawater on a semi industrial pilot plant using different operating conditions. Then, the treatment of seawater, doped with oyster gametes to simulate the filtration of shellfish hatchery effluents, was performed to confirm the hydraulic performance of the air backwash. Indeed, the release of gametes, expulsed by exotic bivalves in the natural environment, could be a risk for the biodiversity preservation. The impact of air backwash on the integrity of oocytes and spermatozoa was identified using flow cytometry and microscopic analyses. When oyster gametes were added, their retention by ultrafiltration was validated. The impact of air backwash on these species viability was a significant information point for the implementation of this process on shellfish production farms.

Development and statistical validation of a quantitative method for the determination of steroid hormones in environmental water by column liquid chromatography/tandem mass spectrometry.

This paper presents an analytical method applied to the determination of 3 natural steroid hormones, estrone, 17 beta-estradiol, and 16 alpha-hydroxyestrone, and the contraceptive estrogen, 17 alpha-ethynylestradiol, at the sub-ng/L level in water samples [surface water and wastewater treatment plant (WWTP) samples]. The solid-phase extraction conditions were optimized using C18 cartridges. Identification and quantification were performed using a column liquid chromatographic/tandem mass spectrometric system with electrospray ionization in the negative mode. Before analyzing steroids in complex matrixes, statistical tools permitted a fine validation of the analytical method, and performance was evaluated for spring water in terms of recovery, specificity, trueness, repeatability, and intralaboratory reproducibility. The results showed the accuracy of the method, and limits of detection (LOD) ranged between 0.02 and 0.21 ng/L. The determination of steroids in WWTP effluents, which contain high levels of organic matter, required an additional purification step on silica cartridges. The high efficiency of the purification was proved with LOD < 0.3 ng/L from 200 mL sample. Specificity of the entire analytical procedure was shown by repeatable recoveries at low and high levels of the calibration range.