Table 2.

Recommended guidelines for research and reporting on phage studies in fish and shellfish. The following information should be considered when designing research studies and should be noted and reported in any publications

Bacterial pathogen under study

• Species, genus, source and accession numbers, when available

• General characteristics

• Known virulence factors (if infecting fish with lab strains)

Fish or shellfish

• Species and common name

• Size, age, and life history stage

• Health status at beginning of experiment (healthy, diseased, immune compromised, etc.)

• Stocking density (for experiment)

Phage characteristics

• Source and characteristics of phage(s) to be used

• Lytic (or lysogenic)

• Phage family (if known): Myoviridae, Siphoviridae, Podoviridae, etc.

• Mixture of phages or single phage to be used in treatment

• Phage titers

Configuration of aquaculture tanks or lab-scale system

• Tank or pond volume and dimensions

• Average depth of pond

• Number of tanks or ponds used for the experiment

Source water

• Source and general quality of water

• Water treatment before use (if applicable), like filtration, UV disinfection, etc.

• Month or season collected and used

Water parameters during experiment

• Range and mean of water temperature, salinity, pH, and dissolved oxygen

• Use of aeration

• Flow rates

• Use of antibiotics (if applicable)

Fish and shellfish challenge

• Route of administering bacterial pathogen(s) to fish or shellfish (natural contamination or through feed, bath, swab, or injection). If injection, indicate site location and how (i.m. or i.p., etc).

• Route of administering phages (via feed, bath, swab, injection (i.m. or i.p., etc.).

• Means of incorporating bacteria or phages into feed, if applicable

• Titer of bacteria added and frequency of addition (if added more than once)

• Titer of phage added and frequency of addition (if added more than once)

• Duration between initial exposure of fish to bacteria and the addition of phages

• Whether treatment is prophylactic, or administered early after infection (before symptoms), or during early or late infection (after symptoms appear)

• Feeding regime: type of feed, amount and frequency administered

• Photoperiod, especially for indoor aquaculture operations or laboratory experiments

• Negative controls used (uninoculated fish and/or fish inoculated with phages only)

• Positive controls (fish inoculated with bacterial pathogen only)

Data collection

• Report the frequency of collection of physical and chemical water quality parameters

• Indicate assay methods used (standard methods, if available) for bacterial and phage testing as well as how frequently tests were conducted

• Report health condition and mortalities of fish at regular intervals, if possible

• Report beginning and final counts of illnesses or mortalities for each experiment.

• Describe the symptoms of ill fish or shellfish

• Report beginning and ending titers of bacteria and phages in fish and water.

Waste product treatment

• Method of treating waste water

• Method of carcass disposal

Quality control

• Know and report the health status of the fish or shellfish before the experiment begins.

• Monitor and report any background levels of target pathogen and any other possible (likely) contaminating pathogens before initiation of experiment and during experiment, as needed

• Report complete methods used for analyses

Data reporting and statistics

• Collect and report data for periods sufficient to show long-term success or failure of phage treatments

• Perform sufficient testing (number of experiments and enough replicates) to make valid statistical claims and report the results

• Provide information on statistical tests performed to evaluate the data

• Disclose all of the above information in papers submitted for publication