Antagonistic activity of Phaeobacter piscinae against the emerging fish pathogen Vibrio crassostreae in aquaculture feed algae

ABSTRACT Aquaculture provides a rich resource of high-quality protein; however, the production is challenged by emerging pathogens such as Vibrio crassostreae. While probiotic bacteria have been proposed as a sustainable solution to reduce pathogen load in aquaculture, their application requires a comprehensive assessment across the aquaculture food chain. The purpose of this study was to determine the antagonistic effect of the potential probiotic bacterium Phaeobacter piscinae against the emerging fish pathogen V. crassostreae in aquaculture feed algae that can be an entry point for pathogens in fish and shellfish aquaculture. P. piscinae strain S26 produces the antibacterial compound tropodithietic acid (TDA). In a plate-based assay, P. piscinae S26 was equally to more effective than the well-studied Phaeobacter inhibens DSM17395 in its inhibition of the fish pathogens Vibrio anguillarum 90-11-286 and V. crassostreae DMC-1. When co-cultured with the microalgae Tetraselmis suecica and Isochrysis galbana, P. piscinae S26 reduced the maximum cell density of V. crassostreae DMC-1 by 2 log and 3–4 log fold, respectively. A TDA-deficient mutant of P. piscinae S26 inhibited V. crassostreae DMC-1 to a lesser extent than the wild type, suggesting that the antagonistic effect involves TDA and other factors. TDA is the prime antagonistic agent of the inhibition of V. anguillarum 90-11-286. Comparative genomics of V. anguillarum 90-11-286 and V. crassostreae DMC-1 revealed that V. crassostreae DMC-1 carries a greater arsenal of antibiotic resistance genes potentially contributing to the reduced effect of TDA. In conclusion, P. piscinae S26 is a promising new candidate for inhibition of emerging pathogens such as V. crassostreae DMC-1 in algal feed systems and could contribute to a more sustainable aquaculture industry. IMPORTANCE The globally important production of fish and shellfish in aquaculture is challenged by disease outbreaks caused by pathogens such as Vibrio crassostreae. These outbreaks not only lead to substantial economic loss and environmental damage, but treatment with antibiotics can also lead to antibiotic resistance affecting human health. Here, we evaluated the potential of probiotic bacteria, specifically the newly identified strain Phaeobacter piscinae S26, to counteract these threats in a sustainable manner. Through a systematic assessment of the antagonistic effect of P. piscinae S26 against V. crassostreae DMC-1, particularly within the context of algal feed systems, the study demonstrates the effectiveness of P. piscinae S26 as probiotic and thereby provides a strategic pathway for addressing disease outbreaks in aquaculture. This finding has the potential of significantly contributing to the long-term stability of the industry, highlighting the potential of probiotics as an efficient and environmentally conscious approach to safeguarding aquaculture productivity against the adverse impact of pathogens.


IMPORTANCE
The globally important production of fish and shellfish in aquaculture is challenged by disease outbreaks caused by pathogens such as Vibrio crassostreae.These outbreaks not only lead to substantial economic loss and environmental damage, but treatment with antibiotics can also lead to antibiotic resistance affecting human health.
Here, we evaluated the potential of probiotic bacteria, specifically the newly identified strain Phaeobacter piscinae S26, to counteract these threats in a sustainable manner.Through a systematic assessment of the antagonistic effect of P. piscinae S26 against V. crassostreae DMC-1, particularly within the context of algal feed systems, the study demonstrates the effectiveness of P. piscinae S26 as probiotic and thereby provides a strategic pathway for addressing disease outbreaks in aquaculture.This finding has the potential of significantly contributing to the long-term stability of the industry, highlighting the potential of probiotics as an efficient and environmentally conscious approach to safeguarding aquaculture productivity against the adverse impact of pathogens.
A quaculture has for decades been a growing industry with 87.5 million tons of high-quality fish and shellfish protein being produced in 2020 (1).However, sustainable production is challenged by the spread of disease caused by fish patho genic bacteria (2)(3)(4).Particularly, species of the Gram-negative gammaproteobacterial Vibrionaceae family are potent pathogens including Vibrio anguillarum, Vibrio harveyi, and Vibrio parahaemolyticus (5,6).Also, Vibrio crassostreae belonging to the Vibrio splendidus group has been identified as an emerging pathogen having caused disease outbreaks and mortalities of several marine aquaculture organisms (7), such as European seabass (8), sea cucumbers (9), Pacific oysters (10)(11)(12), and Yesso scallops (13).In a challenge trial with blue mussel larvae, V. crassostreae killed 73% of challenged mussel larvae after 5 days (14).Pathogenic V. crassostreae strains have been isolated from diseased farmed turbot and European seabass in Norway (15), and from turbot larvae rearing units with high mortality in Norway and Spain (2,16,17).One of the well-stud ied strains, V. crassostreae DMC-1 (previously V. splendidus), was isolated at commercial hatcheries in Galicia, Spain, from the gut of moribund turbot larvae (2,17,18).Fish larvae are a particularly vulnerable stage in the trophic levels of aquaculture because their immature immune system does not render vaccination an effective disease control strategy and they are exposed to fish pathogens via their live feed (19)(20)(21).Current treatments involve the usage of detergents and antibiotics, causing potential environ mental harm and development and spread of antibiotic resistance; probiotic bacteria such as lactic acid bacteria, bacilli, and roseobacters have been proposed as an efficient, sustainable alternative (22)(23)(24)(25).
Members of the marine Gram-negative alphaproteobacterial Roseobacter group including Phaeobacter and Tritonibacter species have been investigated as potential fish probiotics.They are promising candidates for the reduction of fish pathogens in aquaculture (24,(26)(27)(28).They have repeatedly been isolated from aquaculture systems and thus occur naturally in this environment (29,30).They efficiently antagonize fish pathogenic vibrios in direct challenge tests and also in the presence of aquaculture-rel evant biological background such as algae, rotifers, crustaceans, fish eggs, and larvae (26-28, 31, 32).They have neutral or a positive effect on these eukaryotic hosts and a minor effect on the microbiome of the hosts (26,32,33).Several Phaeobacter and Tritonibacter species produce the potent antibacterial agent tropodithietic acid (TDA) (33), which has been linked to the antagonistic activity of Phaeobacter against Vibrio by comparing antibacterial activity to TDA-deficient mutants (34).
The most widely researched roseobacter probiotic candidate is the strain Phaeobacter inhibens DSM17395 (25); however, a novel promising probiotic candidate, Phaeobacter piscinae S26, was isolated from a Greek sea bass larval rearing unit and characterized as belonging to the new Phaeobacter species, P. piscinae (29,35,36).P. piscinae S26 produced the highest concentration of TDA among the tested Phaeobacter strains, including P. inhibens DSM17395, and caused the highest survival of Artemia in Vibrio pathogen trials (27).The majority of fish probiotic studies have used the pathogen V. anguillarum as target organism; however, as outlined above, a range of other vibrios, especially V. crassostreae, are emerging as pathogens in marine larviculture.Using a plate-based assay, Hjelm et al. (18) screened for antagonistic bacteria against V. crassostreae DMC-1 and isolated the strain P. piscinae 27-4.During co-cultivation, P. piscinae 27-4 inhibited V. crassostreae DMC-1 by 3 log units, while in comparison, inhibition of V. anguillarum 90-11-287 was 6-7 log fold.Therefore, the purpose of this study was to assess the effect of the new probiotic candidate, P. piscinae S26, against the fish pathogenic strain, V. crassostreae DMC-1, as a future sustainable biocontrol alternative in aquaculture.We investigated this antagonism in the microalgal systems of Tetraselmis suecica and Isochrysis galbana, as possible targets for probiotic application as these algae are commonly used as live feed in aquaculture.Furthermore, the genome of V. crassostreae DMC-1 was analyzed to suggest possible genotypes for the observed inhibition by P. piscinae S26.

Antagonistic activity of probiotic Phaeobacter against fish pathogenic vibrios
To analyze the antagonistic properties of the new probiotic candidate strain P. piscinae S26 wild type (WT) against the fish pathogens V. crassostreae DMC-1 and V. anguillarum 90-11-286, its activity in plate-based assays was compared to its TDA-deficient mutant S26 ΔtdaB, and the probiotic candidate P. inhibens DSM17395 WT and its TDA-deficient mutant DSM17395 ΔtdaB::GmR (Table 1).Both cell-free supernatants and cell suspen sions of P. piscinae S26 inhibited V. crassostreae DMC-1 (Fig. 1A) and V. anguillarum 90-11-286 (Fig. 1B) in the plate-based assay as shown by halos in the bacterial lawn around the well or inoculum.Both cell-free supernatant and cell suspension of P. piscinae S26 produced inhibition zones of 17 and 19 mm in diameter, respectively, in V. crassos treae DMC-1 lawn.In contrast, P. inhibens DSM17395 produced smaller (8 mm for cell suspension) and no inhibition (cell-free supernatant) on V. crassostreae DMC-1 lawn.The inhibition of V. anguillarum 90-11-286 by P. piscinae S26 and P. inhibens DSM17395 was similarly strong with the inhibition zones of cell-free supernatant and cell suspension of 23 and 21 mm for P. piscinae S26, and 21 and 20 mm for P. inhibens DSM17395, respectively.No inhibition zones, thus, no antibacterial effect was observed for the TDA-deficient mutants of the Phaeobacter strains or the media control.

Antagonistic activity of P. piscinae S26 against the fish pathogenic V. crassostreae DMC-1 in algal systems
Without addition of P. piscinae S26, V. crassostreae DMC-1 grew within 2 days from 5.2 ± 0.7 to 6.2 log CFU/mL ± 0.1 in the I. galbana culture and remained at this cell concentra tion until day 7 (Fig. 2A).Addition of both P. piscinae S26 WT and ΔtdaB inhibited the growth of V. crassostreae DMC-1 throughout the experiment, and the cell concentration remained around the inoculum concentration of 4.5 log CFU/mL (P < 0.0005 after day 0).
P. piscinae S26 WT and ΔtdaB grew in the presence of V. crassostreae DMC-1 in the I. galbana culture from 6.5 ± 0.1 log CFU/mL to 7.5 ± 0.3 log CFU/mL in 7 days (Fig. 3A).The growth of ΔtdaB was delayed as indicated by significantly lower cell concentration of ΔtdaB in comparison to the WT on day 4 (P = 0.003).
In the T. suecica culture, both P. piscinae S26 WT and ΔtdaB grew in the presence of V. crassostreae DMC-1 from 6.1 ± 0.1 and 6.3 ± 0.2 log CFU/mL to 7.2 ± 0.1 and 7.2 ± 0.1 log CFU/mL within 1 day followed by a decline to 6.7 ± 0.1 and 6.5 ± 0.04 log CFU/mL, respectively, on day 8 (Fig. 3B).The growth of the microalgae was generally not affected by the presence of the bacteria.I. galbana and T. suecica grew from 5.1 to 6.8 log cells/mL over 7 days and from 4.5 to 6.0 log cells/mL over 8 days (P > 0.05, except I. galbana axenic control vs P. piscinae S26 WT + V. crassostreae DMC-1 co-culture on day 2, P = 0.02) (Fig. 4).
Finally, as TDA does not appear to be the main driver of V. crassostreae DMC-1 inhibition in algal systems in contrast to the agar-based assay, potential metabolic competition between Vibrio and Phaeobacter was analyzed.V. crassostreae DMC-1 and P. piscinae S26 have unique genomic profiles for the degradation, utilization, and assimilation of-among others-amino acids, aromatic compounds, carbohydrates, and inorganic nutrients (Table 3).P. piscinae S26 has overall 19 unique full metabolic pathways for degradation, while V. crassostreae DMC-1 has only 10.This includes three unique pathways for the degradation of the aromatic compounds anthranilate, methyl salicylate, and salicylate in the P. piscinae S26's genome.Also, P. piscinae S26 has the unique genetic potential to degrade the sulfur-containing organic com pounds dimethylsulfoniopropionate, methanesulfonate, and methyl thiopropionate.Furthermore, a major nutrient source for heterotrophic bacteria in algal systems are

DISCUSSION
With fish pathogens such as vibrios causing significant economic loss to aquaculture systems and the need to prevent antibiotic usage, probiotic bacteria could represent a sustainable solution.For a safe application of such strains, we need to identify the specificity of their activity and test their efficiency in aquaculture-related systems.In this study, we found that the strain P. piscinae S26 is a promising candidate for probiotic application due to its antagonism against vibrios.This effect might even be more pronounced than for the previously tested strain P. inhibens DSM17395 as indicated  (43), ARTS (44), ResFinder (45), RGI, and CARD (46).Presence of antibiotic resistance hits indicated with a '+' , absence with a '-' .Arginine degradation III (arginine decarboxylase/agmatinase pathway) 0.5 1

Glutamate degradation X 1 0
Glutamine degradation I 1 1 Glutamine degradation II by larger inhibition zones in a plate-based inhibition assay.Indeed, Grotkjaer et al. (29) found P. piscinae S26 to produce higher concentrations of TDA than P. inhibens DSM17395, which could drive at least some of this anti-vibrio activity.Vibrios, even within a species, may represent various levels of virulence to aquacul ture organisms and carry a high genetic diversity (3,48,49).Similarly, vibrios have a varying level of sensitivity to the probiotic Phaeobacter and its bioactive compound TDA (25).This was also confirmed for P. piscinae S26 that inhibits both V. anguillarum 90-11-286 and V. crassostreae DMC-1; however, the latter to a lesser extent.For both

Tetrathionate reduction I (to thiosulfate) 0 1
Thiosulfate disproportionation III (rhodanese) 1 1 targets, the activity can be attributed to the production of TDA, as no activity was observed for the TDA-deficient P. piscinae S26 mutants in a plate-based assay.Similarly, Hjelm et al. (18) demonstrated that the inhibitory effect of the TDA-producing strain P. piscinae 27-4 against V. anguillarum 90-11-287 was stronger over time than against V. crassostreae DMC-1 with a 6-log reduction in comparison to a 1-log-fold reduction after 6 days of co-cultivation.When testing the efficacy of P. piscinae S26 to inhibit V. crassostreae DMC-1 in aquaculture-relevant algal cultures, V. crassostreae DMC-1 was reduced by 2 log and 3-4 log fold in I. galbana and T. suecica cultures, respectively.When Grotkjaer et al. (27) tested the activity of P. inhibens DSM17395 against V. anguillarum NB10 in xenic cultures of the algae Dunaliella tertiolecta and T. suecica, the reduction of vibrio was 3 log fold for both systems.Even more pronounced was the effect of P. inhibens DSM17395 against NB10 in a previous study in axenic cultures of T. suecica and Nannochloropsis oculata, which demonstrated a 3-log-fold reduction to complete elimination of the pathogen (26).The authors also observed that NB10 differed in its capability of inhabiting the two different algal systems.Although NB10 colonized T. suecica cells, it could only persist in dense cultures and disappeared from less dense cultures of N. oculata.We observed in our experiments that V. crassostreae DMC-1 would grow to a cell concentration of 6 log CFU/mL; however, although it could maintain this cell concentration in the I. galbana culture, the concentration reduced over time in the T. suecica culture.

Thiosulfate oxidation I (to tetrathionate
Interestingly, both P. piscinae S26 and its TDA-deficient mutant inhibited the growth of V. crassostreae DMC-1 in both algal systems; however, less so for the mutant in the T. suecica culture.Although in direct challenge the inhibitory activity of Phaeobacter could be attributed to the production of TDA, previous work also demonstrated that in aquaculture-relevant systems, TDA is driving the antagonism, but does not fully explain the phenomenon (26).To obtain indications why V. crassostreae DMC-1 appears to be less affected by P. piscinae S26 than V. anguillarum 90-11-286 and why TDA is not the main driver of the antagonistic effect, we analyzed the Vibrio genomes.Although both strains are assigned to the genus Vibrio, they are not closely related and could accordingly have distinct differences in their metabolism, meaning that their overall fitness would be different in algal cultures.They carry a similar biosynthetic potential; however, our analyses demonstrate that V. crassostreae DMC-1 carries a larger arsenal of resistance genes in its genome, highlighting the need to further study this Vibrio species.Although the resistance mechanism to TDA has not been fully elucidated (42), gammaglutamylcyclotransferase activity has been predicted to be involved in Phaeobacter's native resistance.This activity is encoded within both genomes of V. crassostreae DMC-1 and V. anguillarum 90-11-286 and cannot therefore explain the reduced susceptibility of V. crassostreae DMC-1.However, Phaeobacter builds its native resistance by counteracting the TDA-induced proton influx with proton efflux, and our findings demonstrate that V. crassostreae DMC-1 has the greater ability to combat the effect of antibiotics, particularly due to any increased number of efflux pumps in comparison to V. anguillarum 90-11-286.Also, the complex metabolic interactions between the algae and the bacteria could lead to P. piscinae S26 outcompeting V. crassostreae DMC-1 for nutritional resources.The P. piscinae S26 genome carries a greater set of unique degradation, utilization, and assimilation pathways than V. crassostreae DMC-1, which would equip P. piscinae S26 with a broader adaptability to environmental nutrient sources, including those provided by microalgae.A high metabolic versatility is a generally accepted characteristic of bacteria of the Roseobacter group (50,51).Specifically, P. piscinae S26 has the unique genetic potential to degrade the aromatic compounds anthranilate, methyl salicylate, and salicylate, which are involved in defense mechanisms and signaling in plants (52)(53)(54); however, less is known about the production and the role of these compounds by microalgae.Furthermore, Phaeobacter is well known for metabolizing dimethylsulfonio propionate, a sulfur source produced by microalgae (55,56), and an ability that was not found for V. crassostreae DMC-1.Additional genomic analysis identified that P. piscinae S26 and V. crassostreae DMC-1 harbor about a similar number of CAZymes; however, the Phaeobacter genome encodes twofold more GTs than GHs, while it is the other way around for V. crassostreae DMC-1.A diverse set of GTs could allow Phaeobacter to target a wide range of carbohydrates produced by the microalgae and could be important for its adaptation to this specific environment.Furthermore, the production of unknown antibacterial compounds by Phaeobacter could inhibit the growth of V. crassostreae DMC-1 (57)(58)(59)(60)(61).For instance, the algal compound dimethylsulfoniopropio nate has some protective effect against TDA, which has previously been speculated to act as a protectant for eukaryotic hosts (62,63).It is possible that similar effects are present in the systems studied here, but this remains a speculation and would need further investigation in future studies.
In conclusion, the potential of probiotic bacteria to address the economic losses caused by fish pathogens such as vibrios in aquaculture systems and the environmental burden of antibiotic and disinfectant usage holds significant promise for sustainable solutions.This study underscores the importance of specificity and efficacy testing for safe and effective application of probiotic strains.The strain P. piscinae S26 emerges as a strong contender for probiotic use due to its robust antagonistic activity against vibrios, potentially surpassing previously tested strains.Vibrios exhibit diverse levels of virulence and sensitivity to probiotics, which can be influenced by factors such as genetic diversity and metabolic interactions.The role of TDA as a primary driver of the antagonistic effects against vibrios is established, yet the interplay of other factors, such as resistance genes within Vibrio genomes and metabolic competition, demands further investigation.Future studies should investigate the intricate mechanisms underlying these interactions, shedding light on the effectiveness and limitations of probiotics in aquaculture settings, including the effect on algal products such as fatty acid composi tion.This will allow the development of tailored solutions capitalizing on the strengths of probiotics while navigating the complexities of aquatic ecosystems.
Axenic I. galbana CCMP 1323 and T. suecica CCMP 906 were obtained from the Bigelow National Center for Marine Algae and Microbiota (NCMA) and were cultivated in 3% instant ocean (IO; Instant Ocean sea salts; Aquarium Systems, Inc.) with f/2 without silicate (f/2 -Si; NCMA [64]) at 18°C and constant light at ~50 μE m −2 s −1 .Pre-cultures were plated on TSA and MA before each experiment to confirm their axenic status.

Antagonistic activity of probiotic Phaeobacter against fish pathogenic vibrios
The antibacterial activity of P. piscinae S26 WT and ΔtdaB and P. inhibens DSM17395 WT and ΔtdaB::GmR against V. crassostreae DMC-1 and V. anguillarum 90-11-286 was tested using an agar-based assay (65).For preparation of V. crassostreae DMC-1 embedded agar plates (0.1% final concentration of overnight culture), the Petri dishes were placed on ice when pouring the plates as V. crassostreae DMC-1 was very sensitive to the temperature of the agar.Bacterial strains were grown overnight in MB at 25°C and 200 rpm, and either 10 µL of probiotic strain was spotted on top of the Vibrio agar or 50 µL of sterile-filtered supernatant was suspended into wells punched into the Vibrio agar.Inhibition zones were measured after overnight incubation at 25°C.

Antagonistic activity of P. piscinae S26 against the fish pathogenic V. crassostreae DMC-1 in algal systems
To determine the probiotic effect of P. piscinae S26 and its TDA-deficient mutant ΔtdaB against V. crassostreae DMC-1 in algal cultures, four treatments were tested in the algal systems: (i) P. piscinae S26 WT + V. crassostreae DMC-1, (ii) P. piscinae S26 ΔtdaB + V. crassostreae DMC-1, (iii) V. crassostreae DMC-1, and (iv) axenic algae.Cultures were set up in biological triplicates with T. suecica or quadruplicates with I. galbana, resulting in 28 cultures in total with each culture having a volume of 50 mL prepared in a 250-mL Erlenmeyer flask.The estimated starting concentration of the algae was 10 5 algal cells/mL in 3% IO + f/2 -Si medium.V. crassostreae DMC-1 was added to the algal cultures at 0.1% of an overnight culture to an estimated starting concentration of 10 4 Vibrio cells/mL.Either 1% of P. piscinae S26 WT or ΔtdaB was added to an estimated starting concentration of 10 6 Phaeobacter cells/mL.All cultures were incubated at 18°C and constant light at ~50 μE m −2 s −1 , and algal and bacterial concentrations were determined on days 0, 1, 2, 5, and 8 (T.suecica) and 0, 2, 4, and 7 (I.galbana).Bacterial colony forming units were determined after 10-fold serial dilution in 3% IO and plating on TSA (Vibrio CFU) and MA (Phaeobacter CFU).Plates were incubated overnight or for 2-3 days at 25°C, respectively, before counting.For algal cell counts, samples were fixed with 1% formaldehyde and were stored at 4°C until flow cytometry on a Miltenyi MACSQuant VYB.Statistical analysis was performed with an unpaired, two-tailed Student's t-test.

Genomic analysis of V. crassostreae DMC-1
Genomic DNA was extracted from 1 mL of an overnight culture of V. crassostreae DMC-1 in MB using the NucleoSpin tissue kit (740952; Macherey-Nagel).DNA (114 ng/µL) was submitted to Novogene (UK) for 150 bp paired-end sequencing on a NovaSeq Illumina platform.Additionally, long reads were produced on a R9 flow cell using the MinION sequencer (Oxford Nanopore Technologies).Adapters of short reads were removed using AdapterRemoval, and ends were trimmed using fastp.The long reads were trimmed using porechop and were filtered using filtlong.Finally, the short and long reads were assembled using unicycler v0.4.7.The assembly was submitted to NCBI for annotation using the Prokaryotic Genome Annotation Pipeline (PGAP).The genome sequence has been deposited at NCBI under the accession number JAMHIT000000000.BLAST-based average nucleotide identity (ANIb) to V. anguillarum 90-11-286 (Genbank acc.no.GCF_001660505.2) was performed with JSpeciesWS (66).Functional traits encoded in the genomes of V. crassostreae DMC-1 and V. anguillarum 90-11-286 were identified using antiSMASH 7.0.0(43), ARTS (44), and ResFinder 4.1 (database version 2022-05-10) (with default settings of 90% identity threshold and 60% minimum length) (45).Antibiotic resistance genes were predicted with RGI version 5.0.0 and CARD version 3.0.2(46) on the Genoscope platform (47).Metabolic profiles of the V. crassostreae DMC-1 and P. piscinae S26 genomes for degradation, utilization, and assimilation were evaluated with the Metabolic Profile Tool using MicroCyc pathways (67) on the MicroScope platform (47), considering only pathways with a completion level of ≥1.Genomic profiles for carbohydrate degradation of V. crassostreae DMC-1 and P. piscinae S26 (Genbank acc.no.GCF_000826835.1) were analyzed using dbCAN3 (68) with HMMER-and DIAMONDbased searches.Hits were considered for comparison if recognized by both searches.

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) 0 1 Twoa
Completeness of pathway indicated in the range of 0 to 1. Major differences between the strains and the corresponding pathways are highlighted in bold.

TABLE 2
Secondary metabolite gene clusters and antibiotic resistance markers in the genomes of V. crassostreae DMC-1 and V. anguillarum 90-11-286 predicted by antiSMASH

TABLE 3
Metabolic profiles for degradation, utilization, and assimilation encoded in the genomes of P. piscinae S26 and V. crassostreae DMC-1 analyzed with MicroScope (47) a

TABLE 3
Metabolic profiles for degradation, utilization, and assimilation encoded in the genomes of P. piscinae S26 and V. crassostreae DMC-1 analyzed with MicroScope (47) a (Continued)