First report of Perkinsus marinus occurrence associated with wild Pacific oysters Crassostrea gigas from the west coast of Korea.

This study reports the occurrence of Perkinsus marinus associated with wild Pacific oyster (Crassostrea gigas) specimens collected along the west coast of Korea. Confirmation of P. marinus presence was achieved by conventional PCR using World Organization of Animal Health (WOAH)-recommended primers that specifically targeted regions of the rDNA locus (ITS1, 5.8S, and ITS2). Sequencing of 10 samples revealed two distinct sequences differing by a single base pair, indicating potential haplotype variability. One sequence closely resembled the P. marinus strain found in Maryland, USA, whereas the other exhibited divergence, indicative of species diversity in the Korean strain, as was evident from the haplotype network analysis. Further validation involved the Ray's Fluid Thioglycollate Medium (RFTM) assay, which initially yielded inconclusive results, possibly due to low infection intensity. Subsequently, RFTM and 2 M NaOH assays conducted on the isolates in the present study, cultured P. marinus cells in standard DMEM/F12 medium, and a positive P. marinus strain (ATCC 50509), revealed characteristic hypnospores of P. marinus upon Lugol's iodine staining. These comprehensive investigations underscore the conclusive confirmation of P. marinus in Korean waters and mark a significant milestone in our understanding of the distribution and characteristics of this parasite in previously unreported regions.

Impact of dietary piperine on growth performance, immune response, antioxidant status, and immune-related gene expression in pathogen-aggravated Cyprinus carpio.

This study investigated the effects of dietary piperine (PIP) on growth performance, digestive enzymes, serum biochemical parameters, antioxidant and immune responses, and gene expression in Cyprinus carpio challenged with Aeromonas hydrophila. Six diets were prepared with PIP doses of 0, 0.5, 1.0, 2.0, 3.0, and 4.0 g/kg, corresponding with the control, PR50, PR100, PR200, PR300, and PR400, respectively. Fish were challenged with Aeromonas hydrophila after 8 weeks of feeding with the respective diets. Weight gain (PWG) and specific growth rate (SGR) were significantly enhanced, whereas feed conversion ratio (FCR) was lowered in PR200. The cumulative post-challenge survival was improved to 68.43% in the PR200 group compared with 28.08% in the control. Serum total protein and albumin levels were significantly enhanced in the PR200 group compared to the control. However, dietary PIP up to 3 g/kg had no significant effect on serum glucose, cortisol, aspartate aminotransferase, or alkaline phosphatase activities; however, the alanine aminotransferase level was lower (P < 0.05) in the PR200 group than in the control. Intestinal amylase, lipase, and protease activities increased in PR300, and intestinal amylase and lipase increased in the PR100 group (P < 0.05). The serum immunological indices (lysozyme, alternative complement pathway, phagocytic activity, and respiratory burst activity) were higher (P < 0.05) in the PR200 group than in the control group. Serum superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) activities were significantly intensified in PR200-PR300 than in the control group, with the highest activity observed in the PR200 group. Malondialdehyde was significantly lower in the PR200 group than in the control group. Furthermore, SOD, CAT, and Nrf2 expression was strongly upregulated in the liver tissue of the PR200 and PR300 groups compared to that in the control. The transcript levels of pro-inflammatory cytokines viz. IL-1ß and TNF-α were significantly upregulated in the kidneys of the PR100 and PR200 post-challenged. In contrast, the anti-inflammatory cytokine IL-10 was significantly downregulated in the kidneys of PR200. The expression of HSP70 was upregulated only in the PR400. Quadratic regression analysis showed that the optimal dietary PIP level was estimated as 2.07-2.13 g/kg to maximize growth performance. Overall, these results indicate that dietary PIP at an appropriate level can improve immunity, cytokine gene expression, and disease resistance in C. carpio.

Bactericidal efficacy of non-thermal plasma activation against Aeromonas hydrophila and immunological responses of koi (Cyprinus carpio haematopterus).

In the aquaculture industry, an efficient and safe water purification system is important to prevent mass mortality by virulent pathogens. As extensive use of traditional methods (e.g.: povidone-iodine, ozone, ultraviolet irradiation, formalin, and chlorine dioxide) have adverse effects on cultured fish, an appropriate and alternative water purification method is vital for the sustainability of the industry. Non-thermal plasma technology has been successfully used for various biomedical purposes (e.g: food sterilization, medical device disinfection, wound healing, cancer therapy, etc.) and has great potential to be used as a sterilizing system. However, few studies have been conducted on its usefulness in the aquaculture industry. In this study, we investigated the bactericidal efficacy of plasma-activated water induced by non-thermal plasma and its histopathological as well as immunological adverse effects on koi. A highly virulent Aeromonas hydrophila SNU HS7, which caused massive mortality of koi, was used for this study. Non-thermal plasma was applied for 10 min to the fish tanks with 1.2 × 109 CFU/mL SNU HS7 using PLMB-20 system to confirm the sterilization efficacy and to observe the survival and immunological reaction of koi for 14 days. As a result, gross pathological, histopathological, and immunological investigations did not reveal any significant adverse effects in fish as compared to the control groups. To the best of our knowledge, this is the first study showing that non-thermal plasma can be used for sterilization of rearing water without giving significant physiological damage to the fish, even under the assumption of extreme situations. As plasma can effectively sterilize not only bacteria but also other unknown pathogens, the results of this study are showing a promising future in purifying water in aquaculture practice.

Genomic characterization of bacteriophage pSal-SNUABM-01, a novel elongated-head phage infecting Salmonella sp.

Salmonellosis is a disease of critical concern for public health, and the use of bacteriophages is among the most promising approaches to combating Salmonella. As Salmonella has various serotypes and strains, and bacteriophages are virulent to specific hosts, it is important to isolate phages and evaluate interactions with their hosts. In the present study, a novel Salmonella-infecting bacteriophage, pSal-SNUABM-01, was isolated and characterized. Transmission electron microscopy revealed that the bacteriophage is a member of the family Podoviridae and possesses an elongated head and a short tail. The phage genome is circular and 89,500 bp in size. A total of 162 open reading frames were predicted, eight of which were tRNAs. Morphological and genomic analysis revealed that pSal-SNUABM-01 is closely related to phage 7-11. In phylogenetic analysis, pSal-SNUABM-01 and 7-11 did not cluster together with the members of any established genus, suggesting that these two phages comprise a novel genus. The results of this study enhance our understanding of the phylogeny of the family Podoviridae and might be applicable to the development of bacteriophage treatments against Salmonella infections.

Strategy for mass production of lytic Staphylococcus aureus bacteriophage pSa-3: contribution of multiplicity of infection and response surface methodology.

BACKGROUND: Antibiotic-resistant bacteria have emerged as a serious problem; bacteriophages have, therefore, been proposed as a therapeutic alternative to antibiotics. Several authorities, such as pharmacopeia, FDA, have confirmed their safety, and some bacteriophages are commercially available worldwide. The demand for bacteriophages is expected to increase exponentially in the future; hence, there is an urgent need to mass-produce bacteriophages economically. Unlike the replication of non-lytic bacteriophages, lytic bacteriophages are replicated by lysing host bacteria, which leads to the termination of phage production; hence, strategies that can prolong the lysis of host bacteria in bacteria-bacteriophage co-cultures, are required. RESULTS: In the current study, we manipulated the inoculum concentrations of Staphylococcus aureus and phage pSa-3 (multiplicity of infection, MOI), and their energy sources to delay the bactericidal effect while optimizing phage production. We examined an increasing range of bacterial inoculum concentration (2 × 108 to 2 × 109 CFU/mL) to decrease the lag phase, in combination with a decreasing range of phage inoculum (from MOI 0.01 to 0.00000001) to delay the lysis of the host. Bacterial concentration of 2 × 108 CFU/mL and phage MOI of 0.0001 showed the maximum final phage production rate (1.68 × 1010 plaque forming unit (PFU)/mL). With this combination of phage-bacteria inoculum, we selected glycerol, glycine, and calcium as carbon, nitrogen, and divalent ion sources, respectively, for phage production. After optimization using response surface methodology, the final concentration of the lytic Staphylococcus phage was 8.63 × 1010 ± 9.71 × 109 PFU/mL (5.13-fold increase). CONCLUSIONS: Therefore, Staphylococcus phage pSa-3 production can be maximized by increasing the bacterial inoculum and reducing the seeding phage MOI, and this combinatorial strategy could decrease the phage production time. Further, we suggest that response surface methodology has the potential for optimizing the mass production of lytic bacteriophages.

Effectiveness of the guava leaf extracts against lipopolysaccharide-induced oxidative stress and immune responses in Cyprinus carpio.

The anti-inflammatory activity of the guava leaf extracts (GLE) against LPS-induced inflammatory responses in fish macrophage cell lines is well documented. Here, we evaluated the effects of dietary GLE on LPS-induced oxidative stress, immune responses, and glucocorticoid receptor-related gene expression in Cyprinus carpio. Basal diet was supplemented with 0 (control), 100, 150, 200, or 250 mg kg-1 GLE for eight weeks. Highest (p < 0.05) weight gain rate was obtained in fish group supplemented with 200 mg kg-1 of GLE. The results showed that superoxide dismutase, glutathione-S-transferase, glutathione peroxidase, glutathione reductase, lysozyme, and complement C3 decreased, while malondialdehyde level increased in the liver and spleen upon LPS-challenge. Dietary GLE supplementation (especially 200 or 250 mg kg-1) alleviated LPS-induced changes. Similarly, GLE (150-250 mg kg-1) reversed LPS-induced alteration of serum biochemical parameters such as alkaline phosphatase, aspartate transaminase, alanine transaminase, and myeloperoxidase. LPS treatment markedly induced increased the mRNA levels of TNF-α, IL-1ß, and NF-κB p65 in both the liver and kidney tissues; however, GLE pre-treatment attenuated LPS-induced elicitation of TNF-α, IL-ß, and NF-κB p65. Moreover, dietary GLE supplementation significantly increased the expression of HSP70 and HSP90, and glucocorticoid receptor in the liver and kidney after LPS challenge. Thus, GLE attenuated LPS-induced inflammation response by up-regulating glucocorticoid receptor-related gene expression in carp. Finally, GLE supplementation reduced carp mortality after LPS-challenge. These results suggest that dietary supplementation with 200 mg kg-1 GLE is adequate for effectively attenuating LPS-induced oxidative stress and immune-suppressive effects in C. carpio.

Vaccination of fish against Aeromonas hydrophila infections using the novel approach of transcutaneous immunization with dissolving microneedle patches in aquaculture.

The aim of this study was to develop and evaluate a novel route of administration for vaccinating fish against Aeromonas hydrophila infection using a dissolving microneedles (MNs) patch. The A. hydrophila JUNAH strain was inactivated with formalin and used as a vaccine antigen. It was mixed with dissolvable carboxymethyl cellulose (CMC) as the matrix material to produce the MNs patches. When examined with a scanning electron microscope, each patch has 282 uniformly distributed, pyramid-shaped needles on a circular base. In the skin insertion experiment, the MNs patches were confirmed to be capable of penetrating the skin of the fish. Through agglutination assay and analysis of non-specific parameters like lysozyme and superoxide dismutase, it was verified that the antigen embedded into the patch induced adaptive and innate immune responses in the fish. In the challenge experiment, the group inoculated with the MNs patch and the group injected with formalin killed cells (FKC) showed a similar survival rate. Our results suggest that the FKC-loaded MNs patch is a wholly viable method alternative to injection for the vaccination of fish.

Genomic characterization of bacteriophage pEt-SU, a novel phiKZ-related virus infecting Edwardsiella tarda.

A bacteriophage infecting Edwardsiella tarda (named pEt-SU) was isolated from freshwater collected in Chung-ju, South Korea. The whole genome of pEt-SU was 276,734 bp in length, representing the first giant phage infecting Edwardsiella reported to date. A total of 284 putative open reading frames were predicted and annotated. Morphology and genome analyses verified that pEt-SU may be distantly related to the phiKZ-like phages, a well-known giant myovirus. The findings in this study provide new insights into the phages infecting E. tarda ads well as fundamental data for the study of giant phages.

Immunomodulatory Role of Microbial Surfactants, with Special Emphasis on Fish.

Microbial surfactants (biosurfactants) are a broad category of surface-active biomolecules with multifunctional properties. They self-assemble in aqueous solutions and are adsorbed on various interfaces, causing a decrease in surface tension, as well as interfacial tension, solubilization of hydrophobic compounds, and low critical micellization concentrations. Microbial biosurfactants have been investigated and applied in several fields, including bioremediation, biodegradation, food industry, and cosmetics. Biosurfactants also exhibit anti-microbial, anti-biofilm, anti-cancer, anti-inflammatory, wound healing, and immunomodulatory activities. Recently, it has been reported that biosurfactants can increase the immune responses and disease resistance of fish. Among various microbial surfactants, lipopeptides, glycolipids, and phospholipids are predominantly investigated. This review presents the various immunological activities of biosurfactants, mainly glycolipids and lipopeptides. The applications of biosurfactants in aquaculture, as well as their immunomodulatory activities, that make them novel therapeutic candidates have been also discussed in this review.

Pseudomonas tructae sp. nov., novel species isolated from rainbow trout kidney.

This study describes the biochemical and phylogenetic characteristics of a Gram-negative strain, SNU WT1T, isolated from rainbow trout kidney. The 16S rRNA gene sequencing indicated that strain SNU WT1T was highly similar to Pseudomonas wadenswilerensis CCOS 864T and closely related to other Pseudomonas putida-related strains. Multilocus sequence analysis of concatenated partial gyrB, rpoB and rpoD sequences revealed that strain SNU WT1T was distinct from P. putida-related strains and formed a separate clade. The average nucleotide identity and Genome-to-Genome Distance Calculator values were 90.19 and 41.7 %with its closest relative P. wadenswilerensis CCOS 864T; however, it was phenotypically distinct from CCOS 864T with respect to arginine dihydrolase, glucose fermentation, aesculin hydrolysis and N-acetyl-glucosamine assimilation. The major polar lipid of the strain was phosphatidylethanolamine and the major quinone was Q-9. The genome of strain SNU WT1T had 5 685 196 bp with a G+C content of 61.83 mol%. We describe a novel species of genus Pseudomonas, for which the name Pseudomonastructae has been proposed, with SNU WT1T (=KCTC 72265=JCM 33436) as the type strain.

Enhanced bath immersion vaccination through microbubble treatment in the cyprinid loach.

Immunization by bath immersion is likely the simplest method of fish vaccination. Although the route of immunogenicity has not been fully identified, immersion vaccination is clearly a useful labor-saving technique. In this study, microbubble (MB) treatment was assessed for its ability to improve the efficacy of bath immersion vaccination in the cyprinid loach. MBs are commonly defined as minute particles of gas with a diameter of less than 100 µm, which generated free radicals. Here, the efficacy of MB treatment for vaccination enhancement in the cyprinid loach was assessed in direct challenge experiments using the virulent Aeromonas hydrophila JUNAH strain; assessments comprised agglutination titer assay and non-specific parameter analysis. Agglutination titers were high in loaches that were immunized via injection with inactivated cells (FKC group); however, non-specific immune activation parameters (e.g., lysozyme, superoxide dismutase, and phagocytic activity) were more increased in loaches that were immunized via bath immersion with MB treatment. Moreover, MB-treated loaches showed comparable survival rates, relative to loaches immunized via injection with formalin inactivated cells. Thus, higher levels of non-specific immune parameters suggest increased efficacy of this vaccine approach. Improving the effectiveness of bath immersion vaccine will increase its affordability and ease of application in aquaculture.

Immunostimulation of Cyprinus carpio using phage lysate of Aeromonas hydrophila.

Over the last 50 years, various approaches have been established for the development of antigens for immunostimulation. We used phage lysate (PL), composed of inactivated antigens by the lytic bacteriophage pAh 6-c for Aeromonas hydrophila JUNAH strain to develop a vaccine for the prevention of A. hydrophila infection in Cyprinus carpio (common carp). We also assessed the poly D,L lactide-co-glycolic acid (PLGA) microparticles encapsulation method to increase the efficiency of the vaccine. Six groups of vaccines involving encapsulated by PLGA, formalin killed cells, or phage lysate at low or high concentration were prepared for intraperitoneal injection in C. carpio. Blood specimens and head kidney samples were collected at various time points for bacterial agglutination assay and to assess relative expression of immune-related genes interleukin-1 beta (IL-1ß), tumor necrosis factor alpha (TNF-α), lysozyme C, and serum amyloid A (SAA). The vaccine groups using high dose phage lysate antigen showed significantly higher agglutination titers than all other groups at 4- and 6-weeks post vaccination (wpv), with the titer of the PLGA encapsulated vaccine group being highest from 10 wpv to the end of the experiment. The survival rate of fish immunized with the phage lysate vaccines were higher than that of fish immunized with the formailin killed cells vaccine in the challenge experiment conducted 6 wpv. Additionally, the PLGA-encapsulated high dose phage lysate antigen vaccinated groups showed the best protective efficacy in the challenge experiment 12 wpv. Vaccines using the phage lysate antigen also showed higher IL-1ß and lysozyme C gene expression at 7 days post vaccination (dpv) and 2 wpv, and higher TNF-α gene expression was seen at 7 dpv. Higher SAA gene expression was seen in these groups at 1 dpv. These results suggest that phage lysate antigen has the potential to induce robust immune responses than formalin killed cells-based vaccines, and could be more effective as a novel inactivated antigen in preventing A. hydrophila infection in C. carpio.

Application of the bacteriophage pVco-14 to prevent Vibrio coralliilyticus infection in Pacific oyster (Crassostrea gigas) larvae.

Vibrio coralliilyticus infects a variety of shellfish larvae, including Pacific oyster (Crassostrea gigas) larvae worldwide, and remains a major constraint in marine bivalve aquaculture practice, especially in artificial seed production facilities. In this study, we isolated and characterized the bacteriophage (phage) that specifically infects V. coralliilyticus. The phage was designated pVco-14 and classified as Siphoviridae. We also investigated the potential efficacy of the isolated phage against V. coralliilyticus infection. We conducted a survey to replace the overuse of antibiotics, which generate multi-antibiotic-resistant strains and causes environmental pollution. The latent period of pVco-14 was estimated to be approximately 30 min, whereas the burst size was 13.3 PFU/cell. The phage was found to infect four strains of tested V. coralliilyticus. pVco-14 was stable at wide temperature (4-37 °C) and pH (5.0-9.0) ranges. Eighty-one percent of oyster larvae died in an immersion challenge at a dose 1.32 × 105 CFU/ml of virulent V. coralliilyticus (strain 58) within 24 h. When oyster larvae were pre-treated with the phage before the bacterial challenge (bacterial conc.: 1.32 × 104 and 1.32 × 105 CFU/ml), mortality of the phage-treated oyster larvae was lower than that of the untreated control. These results suggest that pVco-14 has potential to be used as a prophylactic agent for preventing V. coralliilyticus infection in marine bivalve hatcheries and can reduce the overuse of antibiotics.

Effect of the Algicide Thiazolidinedione 49 on Immune Responses of Bay Scallop Argopecten Irradians.

The thiazolidinedione 49 (TD49) is an effective algaecide against harmful algae; however, its potential effects on the immune function of the edible bay scallop are unclear. Therefore, the present work studied the effects of TD49 on the immune response in bay scallop by evaluating activities of acid phosphatase (ACP), alkaline phosphatase (ALP), and superoxide dismutase (SOD), as well as nitric oxide (NO) levels, total protein content, and expression of immune genes (CTL-6, PGRP, PrxV, MT, and Cu/Zn-SOD) at 3-48 h post-exposure (hpe) to TD49. The activities of ACP and ALP significantly increased in TD49-treated groups at 3-24 hpe, whereas NO levels decreased significantly in 0.58 and 0.68 µM of TD49 at 6-24 hpe, after which the level was similar to that in the untreated control. Moreover, SOD activity significantly increased in all three concentration groups at 3-6 hpe, while it decreased at 12 hpe in the 0.68 µM TD49 treatment group. Notably, total protein content increased with TD49 treatment at each time interval. The results revealed that variable effects on the expression of immune-related genes were observed after treatment with TD49. The findings demonstrate that exposure of scallops to TD49 changes immune responses and expression of immune-related genes. We hypothesize that TD49 may disrupt immune system in bay scallop. The current investigation highlights the potential negative effects of using TD49 as an algaecide on marine economic bivalves to control harmful algal blooms in marine environments.

Phage Application for the Protection from Acute Hepatopancreatic Necrosis Disease (AHPND) in Penaeus vannamei.

Acute hepatopancreatic necrosis disease (AHPND) caused by Vibrio parahaemolyticus has been one of the most problematic diseases in marine shrimp aquaculture throughout Southeast Asia and Latin America. To evaluate the effectiveness of a bacteriophage (phage) treatment for AHPND, a series of bioassays were carried out in a marine shrimp (Penaeus vannamei) model using an AHPND-V. parahaemolyticus strain that is highly pathogenic to shrimp. We monitored the mortality and histopathological changes during phage treatment. Shrimps treated with phage prophylaxis and phage therapy displayed significant protection from AHPND and survived a lethal bacterial challenge.

Effects of algal toxin okadaic acid on the non-specific immune and antioxidant response of bay scallop (Argopecten irradians).

Okadaic acid (OA) is produced by dinoflagellates during harmful algal blooms and is a diarrhetic shellfish-poisoning (DSP) toxin. This toxin is particularly problematic for bivalves that are cultured for human consumption. This study aimed to reveal the effects of exposure to OA on the non-specific immune responses of bay scallop, Argopecten irradians. Various immunological parameters (superoxide dismutase (SOD), acid phosphatase (ACP), alkaline phosphatase (ALP), lysozyme activities, and total protein level) were assessed in the hemolymph of bay scallops at 3, 6, 12, 24, and 48 h post-exposure (hpe) to different concentrations (50, 100, and 500 nM) of OA. Moreover, the expression of immune system-related genes (MnSOD, PrxV, PGRP, and BD) was also measured. Results showed that SOD and ACP activities were decreased between 12 and 48 hpe. The ALP, lysozyme activities, and total protein levels were also modulated after exposure to different concentrations of OA. The expression of immune-system-related genes was also assessed at different time points during the exposure period. Overall, our results suggest that the exposure to OA had negative effects on the antioxidant and non-specific immune responses, and even disrupted the metabolism of bay scallops, making them more vulnerable to environmental stress-inducing agents; they provide a better understanding of the response status of bivalves against DSP toxins.

Immune response of the bay scallop, Argopecten irradians, after exposure to the algicide palmitoleic acid.

Palmitoleic acid (PA) is an effective algicide against the toxin-producing dinoflagellate Alexandrium tamarense; however, its effects on the immune system of the edible bay scallop Argopecten irradians are unclear. Therefore, we investigated the effects of PA on the immune response in A. irradians by assessing total haemocyte counts (THC), alkaline phosphatase activity (ALP), nitrite oxide (NO), glutathione (GSH), and lactate dehydrogenase (LDH) levels, as well as the expression of immune-related genes (FREP, PGRP, HSP90, MnSOD, and Cu/ZnSOD) at various hours post-exposure (hpe) to the compound. THC decreased in PA-treated groups, whereas ALP increased significantly in all of the PA treatment groups at 3 hpe, after which it significantly decreased. The LDH and NO levels were significantly enhanced in the high and medium concentration group. Notably, the GSH level increased in all PA treatment groups at each time interval. Our study revealed that after treatment with different concentrations of PA, variable effects on the expression of genes involved in the immune system response were observed. The results of our study demonstrate that immersing scallops in PA at effective concentrations could result in differential effects on immune system responses and expression of immune-related genes. Specifically, PA may disrupt the endocrine system or affect signal transduction pathways in the scallops. Therefore, the present study highlights the potential risk of using the PA as an algicide to control algal bloom outbreaks in the marine environment.

Isolation and Comparative Genomic Analysis of T1-Like Shigella Bacteriophage pSf-2.

The increasing prevalence of antibiotic-resistant Shigella sp. emphasizes that alternatives to conventional antibiotics are needed. Siphoviridae bacteriophage (phage), pSf-2, infecting S. flexneri ATCC(®) 12022 was isolated from Geolpocheon stream in Korea. Morphological analysis by transmission electron microscopy revealed that pSf-2 has a head of about 57 ± 4 nm in diameter with a long tail of 136 ± 3 nm in length and 15 ± 2 nm in width. One-step growth analysis revealed that pSf-2 has latent period of 30 min and burst size of 16 PFU/infected cell. The DNA genome of pSf-2 is composed of 50,109 bp with a G+C content of 45.44 %. The genome encodes 83 putative ORFs, 19 putative promoters, and 23 transcriptional terminator regions. Genome sequence analysis of pSf-2 and comparative analysis with the homologous T1-like Shigella phages, Shfl1 and pSf-1, revealed that pSf-2 is a novel T1-like Shigella phage. These results showed that pSf-2 might have a high potential as a biocontrol agent to control shigellosis. Also, the genomic information may lead to further understanding of phage biodiversity, especially T1-like phages.

Effect of the Algaecide Palmitoleic Acid on the Immune Function of the Bay Scallop Argopecten irradians.

Palmitoleic acid (PA), an algicidal compound, is used against the toxin producing dinofagelate Alexandrium tamarense, however, its impact on the edible bay scallop (Argopecten irradians) is still unclear. Therefore, we investigated the impacts of effective algicidal concentrations (20, 40, and 80 mg/L) of PA on immune responses in A. irradians. Various immune parameters including acid phosphatase (ACP) activity, superoxide dismutase (SOD), lysozyme, phagocytic activity, total protein, malondialdehyde (MDA) level, and reactive oxygen species (ROS) production and the expression of immune-related genes (PrxV, CLT-6, MT, and BD) were measured at 3, 6, 12, 24, and 48 h post-exposure (hpe) to PA. Lysozyme activity was lower in scallops at 12-48 hpe to 80 mg/L. SOD, ACP activity, ROS production, the total protein, and MDA level was higher at 12 to 48 hpe with different concentrations of PA. Phagocytic activity increased at 6-12 hpe to 40-80 mg/L of PA, but decreased at 24-48 hpe. The expressions of genes PrxV, CLT-6, MT and BD down-regulated at 3 hpe were observed, while differential expressions from 6-48 hpe with different concentrations of PA. The present study demonstrated that immersing A. irradians in PA at effective concentrations could result in differential effects on non-specific immune responses and expressions of immune-related genes.

Marine Toxin Okadaic Acid Affects the Immune Function of Bay Scallop (Argopecten irradians).

Okadaic acid (OA) is produced by dinoflagellates during harmful algal blooms and is a diarrhetic shellfish poisoning toxin. This toxin is particularly problematic for bivalves that are cultured for human consumption. This study aimed to reveal the effects of exposure to OA on the immune responses of bay scallop, Argopecten irradians. Various immunological parameters were assessed (total hemocyte counts (THC), reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), lactate dehydrogenase (LDH), and nitric oxide (NO) in the hemolymph of scallops at 3, 6, 12, 24, and 48 h post-exposure (hpe) to different concentrations of OA (50, 100, and 500 nM). Moreover, the expression of immune-system-related genes (CLT-6, FREP, HSP90, MT, and Cu/ZnSOD) was also measured. Results showed that ROS, MDA, and NO levels and LDH activity were enhanced after exposure to different concentrations of OA; however, both THC and GSH decreased between 24-48 hpe. The expression of immune-system-related genes was also assessed at different time points during the exposure period. Overall, our results suggest that exposure to OA had negative effects on immune system function, increased oxygenic stress, and disrupted metabolism of bay scallops.