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Harikrishnan, Ramasamy,Balasundaram, Chellam,Moon, Young-Gun,Kim, Man-Chul,Kim, Ju-Sang,Dharaneedharan, Subramanian,Heo, Moon-Soo Akadémiai Kiadó 2010 ACTA VETERINARIA HUNGARICA Vol.58 No.1
<P>Goldfish ( Carassius auratus ) weighing 13 +/- 2 g were administered intramuscularly a sublethal dose (1.8 x 10 3 cfu/ml) of Aeromonas hydrophila to induce ulcerative dermatitis. On day 3 and day 6 after infection the fish were dip-treated (for 5 min/day) with a tri-herbal concoction of Curcuma longa, Ocimum sanctum and Azadirachta indica (1%). The LD 50 value was recorded at a concentration of 2.3 x 10 4 between 30 and 36 days after infection. In the infected untreated group the cumulative mortality was higher, while in the early-treated group (day 3) there was no mortality. In the late-treated group (6th day) the mortality increased to 23.3% on day 36. In the infected group the size of ulcers progressively increased from 43.3% of the body length on day 18 to 86.7% on day 36. In the early-treated fish the size of ulcers was 23.3% of the body length on day 18; by day 36 after treatment the ulcer had completely healed. In contrast, there was only a moderate recovery in the late-treated group. From the results it can be concluded that early tri-herbal treatment ensures successful recovery from ulcerative dermatitis induced by A. hydrophila .</P>
Harikrishnan, Ramasamy,Balasundaram, Chellam,Heo, Moon-Soo Elsevier 2010 FISH AND SHELLFISH IMMUNOLOGY Vol.29 No.1
<P><B>Abstract</B></P><P>Scuticociliatosis caused by about 20 species belonging to the Phylum Ciliophora has been recognized as an emerging problem inflicting significant economic loss in aquaculture industry in the world. Among these <I>Philasterides dicentrarchi</I>, <I>Miamiensis avidus</I>, and <I>Uronema marinum</I> are the three species responsible for scuticociliatosis in olive flounder farms of South Korea. Some of the parasites living or scavenger ciliates also have become parasites of aquaculture fish. The major clinico-pathological manifestations of scuticociliatosis infected fishes are anemia, weight loss, dark coloration, enteritis, excessive body mucus, yellowish intestinal mucus, loss of scales, hemorrhagic and/or bleached spots on the skin, and dermal necrotic lesions that finally destroy tissues lead to high mortalities. Affected fish exhibit organ-specific pathological changes in the brain, eyes, muscle, gills, liver, kidney, intestine, and stomach that lead to severe mortality. At present, farmers in South Korea manage scuticociliatosis by using therapeutic measures, such as application of antibiotics like oxytetracycline, gentamycine, tetracycline, amoxycililin, and cefazolin and chemicals, such as formalin, hydrogen peroxide, malachite green, and jenoclean at a concentration of 350 ± 150 ppm. However till date, no systematic scientific study has been conducted under field condition on the efficacy of these management measures. Under laboratory condition the ciliate can be effectively controlled with the antibiotics and chemicals while on the host, but on entering the host no systemic chemotherapeutic treatment has been yet proven effective. Furthermore the indiscriminate uses of harmful chemicals in aquaculture are increasingly becoming a cause of concern. Recently formalin and malachite green, the most widely used chemicals have been banned in food fish production by FDA as not consumer friendly and being carcinogenic respectively. Vaccines and immunostimulants can induce good immune response and protect against scuticociliatosis as it has been proved in the case of freshwater Ich. Now a days a number of probiotics and herbal formulations are in use against freshwater bacterial and fungal diseases, while, little information is available regarding the different prophylactic measures against marine scuticociliatosis. This review attempts to provide information on the various prophylaxic measures practiced against scuticociliatosis with special reference to olive flounder farms in South Korea.</P>
Harikrishnan, Ramasamy,Heo, Jaehyun,Balasundaram, Chellam,Kim, Man-Chul,Kim, Ju-Sang,Han, Yong-Jae,Heo, Moon-Soo Elsevier 2010 Veterinary parasitology Vol.170 No.1
<P><B>Abstract</B></P><P>We report the effect of aqueous-, ethanol- and methanol-solvent-derived extracts of three traditional Korean herbs, <I>Punica granatum</I>, <I>Chrysanthemum cinerariaefolium</I> and <I>Zanthoxylum schinifolium</I>, by monitoring the innate immune mechanisms, such as phagocytosis activity, respiratory burst activity, alternative complement activity and lysozyme activity and the functional immunity in terms of percentage mortality and relative percent survival (RPS) in olive flounder (<I>Paralichthys olivaceus</I>) against <I>Uronema marinum</I> (1×10<SUP>5</SUP>ciliatesml<SUP>−1</SUP>) for 30 days. Fish were intraperitoneally administered with 5, 50 and 100mgkg<SUP>−1</SUP> body weight of each traditional Korean medicinal (TKM) solvent extract except the control and infected untreated groups. In all the treated groups at concentrations of 50 and 100mgkg<SUP>−1</SUP> body weight, the chosen innate immune parameters were found significantly enhanced when compared to 0mgkg<SUP>−1</SUP> dose. However, at 5mgkg<SUP>−1</SUP> the tested immune parameters did not vary. Administration of TKM solvent extracts preceding the challenge with <I>U. marinum</I> for 30 days significantly reduced the percentage mortality with the consequent increase in RPS. Administration of 50 and 100mgkg<SUP>−1</SUP> TKM solvent extracts clearly enhanced the innate immune responses and disease resistance in <I>P. olivaceus</I> against <I>U. marinum</I>.</P>
Harikrishnan, Ramasamy,Balasundaram, Chellam,Kim, Man-Chul,Kim, Ju-Sang,Han, Yong-Jae,Heo, Moon-Soo Wiley (John WileySons) 2010 Journal of aquatic animal health Vol.22 No.4
<P>We investigated the effect of a mixed herb-enriched diet obtained from pomegranate Punica granatum, Dalmatian chrysanthemum Chrysanthemum cinerariaefolium, and mastic-leaved prickly-ash Zanthoxylum schinifolium on innate immune mechanisms (e.g., phagocytosis activity, respiratory burst activity, alternative complement activity, lysozyme activity, and disease resistance) of olive flounder Paralichthys olivaceus against the scuticociliate Philasterides dicentrarchi. All experimental groups were challenged with P. dicentrarchi (1 x 10(5) ciliates/mL) through intraperitoneal administration of the pathogen (50 microL) on day 1. On day 7, the infected groups were fed 0, 5, 50, and 100 mg/kg of the enriched diets. The innate immune parameters, cumulative mortality, and relative percent survival (RPS) were assessed at weeks 1, 2, 3, and 4. Administration of 50 or 100 mg/kg of the herbal-enriched diet enhanced immunity throughout the experimental period. However, at the 5-mg/kg dosage, the enriched diet did not enhance the innate immune estimates at any time. At doses of 50 and 100 mg/kg, administration of the diet preceding the challenge with P. dicentrarchi decreased the percentage cumulative mortality in the experimental groups and thereby increased RPS values. This study reports that administration of 50 or 100 mg/kg mixed herbal-enriched diet could positively influence the innate immune response to P. dicentrarchi and enhance the health status of olive flounder with respect to this microbe.</P>
Harikrishnan, Ramasamy,Balasundaram, Chellam,Heo, Moon-Soo Elsevier 2010 FISH AND SHELLFISH IMMUNOLOGY Vol.29 No.6
<P><B>Abstract</B></P><P>The effect of <I>Lactobacillus sakei</I> BK19 (10<SUP>8</SUP> cells g<SUP>−1</SUP>) supplemented diet fed to kelp grouper, <I>Epinephelus bruneus</I> against streptococcosis caused by <I>Streptococcus iniae</I> and <I>Streptococcus parauberis</I> with reference to the innate immune response and disease resistance was evaluated at 1, 2, and 4 weeks. Maximum reduction in mortalities was observed in kelper feeding the probiotic diet for two weeks after challenged with the pathogens when compared to the infected group fed with basal diet; similarly the cellular and humoral immune responses such as head kidney macrophage phagocytic and peroxidase activities, serum lysozyme activity, and total protein levels increased significantly. The results reveal that, in streptococcosis infected kelp grouper feeding <I>L. sakei</I> BK19 enriched diet affords a higher level of disease protection due to stimulation of immune system.</P>
Harikrishnan, Ramasamy,Kim, Ju-Sang,Balasundaram, Chellam,Heo, Moon-Soo The Korean Society of Fish Pathology 2008 한국어병학회지 Vol.21 No.2
Goldfish, Carassius auratus (wt 13 g) was intramuscularly infected with Aeromonas hydrophila (4.3x106 cfu / ml). Infected gills showed edematous lamellae with bacterial invasion into the capillaries and gill congestion on 12th day. By 24th day post-infection, histological analysis revealed irregular aggregates of macrophages in gill lamellae, large amount of mucus cells, gill lamellae edematous with bacterial invasion into capillaries, gill congestion and damaged gill epithelium with hyperplasia. Inflammation of the gill filament and hemorrhage globe was associated with the development of severe necrosis on the 36th day in the infected fishes. In infected and herbal treated fish the regenerative responses like fibrosis and infiltration of the leucocytes (neutrophils and monocytes) occurred on 12th day; moderate hypertrophy in the gills was noticed on the 36th day. These results suggest that phytotherapy ensures better protection and regenerative response against A. hydrophila infection in goldfish, C. auratus.
Harikrishnan, Ramasamy,Kim, Ju-Sang,Kim, Man-Chul,Dharaneedharan, Subramanian,Kim, Dong-Hwi,Hong, Seung-Hyun,Song, Chang-Young,Balasundaram, Chellam,Heo, Moon-Soo Elsevier 2012 Experimental parasitology Vol.131 No.2
<P><B>Graphical abstract</B></P><P>The cumulative mortality in groups fed control (C) and <I>S. maritima</I> extract supplementation diets at 0%, 0.01%, 0.1%, and 1.0% and challenged with <I>M. avidus</I>.<ce:figure id='f0050'></ce:figure></P><P><B>Highlights</B></P><P>► RBC significantly increased with 0.1% and 1.0% enriched diets on week 4. ► Hb and Ht levels increased when with 0.1% and 1.0% diets on weeks 2 and 4. ► Lym, Mon, Neu, TP, GLU, and CAL levels increased in 0.1% and 1.0% diets on weeks 2 and 4. ► Lysozyme activity enhanced in 0.1% and 1.0% diets from weeks 1 to 4. ► Scuticocidal and respiratory burst activities enhanced in 0.1% and 1.0% diets on weeks 2 and 4.</P> <P><B>Abstract</B></P><P>The effect of <I>Suaeda maritima</I> enriched diet on blood physiology, innate immune response, and disease resistance in olive flounder <I>Paralichythys olivaceus</I> against <I>Miamiensis avidus</I> on weeks 1, 2, and 4 was investigated. Feeding with any enriched diet and then challenging with <I>M. avidus</I> significantly increased white blood cells (WBC) on weeks 2 and 4; the red blood cells (RBC) significantly increased with 0.1% and 1.0% enriched diets on week 4. The hemoglobin (Hb) and hematocrit (Ht) levels significantly increased when fed with 0.1% and 1.0% supplementation diets on weeks 2 and 4. The mean corpuscular volume (MCV) did not significantly vary with any diet and time; however the mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) significantly increased with 0.1% and 1.0% supplementation diets on weeks 2 and 4. The leukocytes such as lymphocytes (Lym), monocytes (Mon), neutrophils (Neu) and biochemical parameters such as total protein (TP), glucose (GLU), and calcium (CAL) levels significantly increased in 0.1% and 1.0% supplementation diet fed groups on weeks 2 and 4. The serum lysozyme activity was significantly enhanced in 0.1% and 1.0% supplementation diet fed groups from weeks 1 to 4 when compared to the control (0% herbal extract enriched diet). The scuticocidal activity and respiratory burst activity were significantly enhanced when fish were fed with 0.1% and 1.0% supplementation diets from weeks 2 and 4. The protective effect in terms of cumulative mortality (50% and 40%) was low in groups on being fed with 0.1% and 1.0% supplemented diet. Therefore the present study suggested that 0.1% and 1.0% <I>S. maritime</I>-supplemented diets protect the hematological and biochemical parameters, improving the innate immunity, affording protection disease from <I>M. avidus</I> infection in olive flounder.</P>