Impact of low-dose ozone nanobubble treatments on antimicrobial resistance genes in pond water.

Antimicrobial resistance (AMR) poses a significant global health threat as the silent pandemic. Because of the use of antimicrobials in aquaculture systems, fish farms may be potential reservoirs for the dissemination of antimicrobial resistance genes (ARGs). Treatments with disinfectants have been promoted to reduce the use of antibiotics; however, the effect of these types of treatments on AMR or ARGs is not well known. This study aimed to evaluate the effects of low dose ozone treatments (0.15 mg/L) on ARG dynamics in pond water using metagenomic shotgun sequencing analysis. The results suggested that ozone disinfection can increase the relative abundance of acquired ARGs and intrinsic efflux mediated ARGs found in the resistance nodulation cell division (RND) family. Notably, a co-occurrence of efflux and non-efflux ARGs within the same bacterial genera was also observed, with most of these genera dominating the bacterial population following ozone treatments. These findings suggest that ozone treatments may selectively favor the survival of bacterial genera harboring efflux ARGs, which may also have non-efflux ARGs. This study underscores the importance of considering the potential impacts of disinfection practices on AMR gene dissemination particularly in aquaculture settings where disinfectants are frequently used at low levels. Future endeavors should prioritize the evaluation of these strategies, as they may be associated with an increased risk of AMR in aquatic environments.

In situ generation of highly localized chlorine by laser-induced graphene electrodes during electrochemical disinfection.

Laser-induced graphene (LIG) has gained popularity for electrochemical water disinfection due to its efficient antimicrobial activity when activated with low voltages. However, the antimicrobial mechanism of LIG electrodes is not yet fully understood. This study demonstrated an array of mechanisms working synergistically to inactivate bacteria during electrochemical treatment using LIG electrodes, including the generation of oxidants, changes in pH-specifically high alkalinity associated with the cathode, and electro-adsorption on the electrodes. All these mechanisms may contribute to the disinfection process when bacteria are close to the surface of the electrodes where inactivation was independent of the reactive chlorine species (RCS); however, RCS was likely responsible for the predominant cause of antibacterial effects in the bulk solution (i.e., ≥100 mL in our study). Furthermore, the concentration and diffusion kinetics of RCS in solution was voltage-dependent. At 6 V, RCS achieved a high concentration in water, while at 3 V, RCS was highly localized on the LIG surface but not measurable in water. Despite this, the LIG electrodes activated by 3 V achieved a 5.5-log reduction in Escherichia coli (E.coli) after 120-min electrolysis without detectable chlorine, chlorate, or perchlorate in the water, suggesting a promising system for efficient, energy-saving, and safe electro-disinfection.

Establishment of a real-time Recombinase Polymerase Amplification (RPA) for the detection of decapod iridescent virus 1 (DIV1).

A rapid and simple real-time recombinase polymerase amplification (RPA) assay was developed to detect decapod iridescent virus 1 (DIV1). The assay was developed using optimized primers and probes designed from the conserved sequence of the DIV1 major capsid protein (MCP) gene. Using the optimized RPA assay, the DIV1 test was completed within 20 min at 39 ℃. The RPA assay was specific to DIV1 with a detection limit of 2.3 × 101 copies/reaction and there was no cross-reactivity with the other aquatic pathogens (WSSV, IHHNV, NHPB, VpAHPND, EHP, IMNV, YHV-1 and GAV) tested. Four out of 45 field-collected shrimp samples tested positive for DIV1 by real-time RPA. The same assay results were obtained by both methods. Thus, the real-time RPA assay developed could be a simple, rapid, sensitive, reliable and affordable method for the on-site diagnosis of DIV1 infection and has significant potential in helping to control DIV1 infections and reduce economic losses to the shrimp industry.

Copper/Carbon Core/Shell Nanoparticles: A Potential Material to Control the Fish Pathogen Saprolegnia parasitica.

Copper-based fungicides have a long history of usage in agriculture and aquaculture. With the rapid development of metal-based nanoparticles, copper-based nanoparticles have attracted attention as a potential material for prevention and control of Saprolegnia parasitica. The present study investigated the effectiveness of copper/carbon core/shell nanoparticles (CCCSNs) and a commercial CCCSNs filter product (COPPERWARE®) against S. parasitica in a recirculating system. Results showed that the growth of agar plugs with mycelium was significantly suppressed after exposure to both CCCSNs powder and COPPERWARE® filters. Even the lowest concentration of CCCSNs used in our study (i.e., 100 mg/mL) exhibited significant inhibitory effects on S. parasitica. The smallest quantity of the filter product COPPERWARE® (3.75 × 3.7 × 1.2 cm, 2.58 g) used in our aquarium study also demonstrated significant inhibition compared with the control group. However, we observed leaching of copper into the water especially when larger quantities of COPPERWARE® were used. Water turbidity issues were also observed in tanks with the filter material. Besides these issues, which should be further investigated if the product is to be used on aquatic species sensitive to copper, CCCSNs has promising potential for water disinfection.

Fraudulent antibiotic products on the market for aquaculture use.

Antibiotics in aquaculture are used to treat bacterial infections. In order for these products to work effectively fish need to be properly dosed. One of the emerging issues in aquaculture is under-dosing large populations of fish with antibiotics. This happens inadvertently for a number of reasons including the use of fraudulent medications. In this study we evaluated 17 antibiotic products (8 florfenicol and 9 oxytetracycline brands purchased in Asia) by HPLC to determine if the product labels accurately reflected the active pharmaceutical ingredient (API) in the package. We determined authenticity scores for different batches of products at two separate laboratories by comparing the observed API to the label API concentration. We found that 48 % of the antibiotic batches had authenticity scores below 80 % (i.e. observed API in package was at least 20 % less than the label API concentration). Further, there were 9 or the 31 batches of drugs tested had no measureable API. Some products had variation in their authenticity scores between batches making it difficult to rely on a brand. The price of florfenicol products may help identify products with low authenticity scores, but in the case of oxytetracycline, the price of all the products tested was relatively similar. The findings in this study suggest that not all florfenicol and oxytetracycline antibiotic products on the market in Asia have API concentrations indicated on their labels. This could be problematic for medicating fish on aquaculture farms.