Antiparasitic activity of the medicinal plant Dillenia suffruticosa against the marine leech Zeylanicobdella arugamensis (Hirudinea) and its phytochemical composition

Zeylanicobdella arugamensis (Annelida: Hirudinea), a marine parasitic leech, is currently affecting different species of cultured groupers, hybrid groupers, snappers and sea bass in Malaysia. Dillenia suffruticosa (Dilleniaceae), a medicinal plant found in Sabah, has been selected in our experiment to kill the leeches as a natural control method. The leech‐infested hybrid groupers were collected from aquaculture facilities, and the isolated leeches were challenged against methanol extract of D. suffruticosa leaves. The experiment was carried out using various concentrations of the extracts such as 25, 50 and 100 mg/ml. The methanol extract showed significant antiparasitic activity against Z. arugamensis with 100% mortality at a concentration of 100 mg/ml in 14.39 ± 3.75 min., followed by 50 and 25 mg/ml in 32.97 ± 9.29 and 41.77 ± 5.40 min., respectively. The phytochemical composition of the extract was determined using GC‐MS analysis to understand the nature of the principal compounds responsible for its antiparasitic properties. The leaves of D. suffruticosa demonstrated the presence of different bioactive compounds of various natures with varying percentages. Thus, it could be revealed that the methanol extract of D. suffruticosa mainly contains vital phytochemical compounds and showed an effective antiparasitic activity against the harmful leeches of hybrid groupers.     

Characterization of triploid hybrid groupers from interspecies hybridization (Epinephelus coioides ♀ × Epinephelus lanceolatus ♂)

Interspecies hybridization is widely used in aquaculture as a beneficial strategy. Diploid and triploid hybrids have been detected from the interspecies hybridization of Epinephelus coioides ♀ × Epinephelus lanceolatus ♂. This is the first report of triploidization through hybridization in grouper. Confirmation has been obtained through flow cytometry, karyotyping and erythrocyte nuclei measurement. The chromosome numbers of E. coioides, E. lanceolatus, diploid hybrid grouper are 48 and triploid hybrid grouper are 72. Measurements of erythrocyte nuclei indicate that triploid fish have a larger nuclear surface than the diploid groupers, and the average ratio of triploid to diploid surface area is 1.59. During the first 1.5 years, triploid hybrid groupers grow faster than diploid hybrid groupers or either parent species. The average growth rate of triploid hybrids is 1.61 times greater than that of diploid hybrids at 6 months of age and 1.43 times greater at 18 months of age. The triploid hybrid groupers are inferior in gonadal development, with no primary‐growth‐stage oocytes appearing in the gonads at 18 months of age. Morphological studies indicate that triploid hybrid groupers have distinctive differences in snout length, eye diameter, body trunk shape, and tail shape development compared with diploid hybrid groupers. Triploid hybrid groupers have an advantage in growth ability, and artificial breeding of triploid groupers might be of great potential use in the grouper aquaculture industry.     

Effect of salinity and temperature on marine leech,Zeylanicobdella arugamensis (De Silva) under laboratory conditions

The high prevalence (80–100%) of the marine leech Zeylanicobdella arugamensis (De Silva) on cage‐cultured Asian sea bass Lates calcarifer (Bloch) led us to investigate the percentage of juvenile leeches hatched from deposited cocoons, survival of juvenile and adult marine leeches at different salinity and temperature. The results showed that the hatching percentage of juvenile leeches was highest at salinity of 30 ppt (32.5 ± 2.8%) followed by 20 ppt (18.0 ± 4.3%) and 10 ppt (12.1 ± 1.4%), respectively. It was found that the adult and juvenile leeches could live up to an average range of 4–7 days at salinity ranging from 10 to 40 ppt. The juvenile leeches were able to hatch at temperature ranging from 25 to 35 °C but unable to hatch at 40 °C. The survival period of adult and juvenile leeches ranged from 11 to 16 days at 25 °C, which was comparatively longer than 5–13 days and 10 h – 5 days at 27–30 °C and 35–40 °C, respectively. The study provided the information on the physical parameters of salinity and temperature which are most optimal for the marine leech Z. arugamensis to propagate.     

Isolation,identification and pathogenicity of Vibrio harveyi,the causal agent of skin ulcer disease in juvenile hybrid groupers Epinephelus fuscoguttatus × Epinephelus lanceolatus

The hybrid grouper, Epinephelus fuscoguttatus (♀) × Epinephelus lanceolatus (♂), is a newly bred cultivated marine fish species of high economic value. However, a skin ulcer disease with high mortality has occurred, and the responsible pathogen remains unknown. In this study, we summarized the epidemic status and external signs of this disease. We screened potential pathogens and finally isolated one bacterial strain ML01 from affected fish. We subjected healthy juvenile hybrid groupers to bacterial challenge tests with the isolate by immersion, immersion after dermal abrasion and intraperitoneal injection, respectively. Within 14 days post‐infection, the isolate ML01 caused mass mortality of juveniles infected via immersion after dermal abrasion or intraperitoneal injection. Diseased juveniles displayed obvious signs of skin ulcers. The median lethal dose of ML01 by intraperitoneal injection was 1.10 × 10⁵ colony‐forming units. ML01 was identified as Vibrio harveyi by bacterial morphology, analytical profile index identification, 16S rDNA sequencing and multilocus sequence analysis. Antibiotic susceptibility tests showed that ML01 was sensitive to ceftriaxone, doxycycline and minocycline. The results of this study suggest that V. harveyi is the causal agent of skin ulcer disease in juvenile hybrid groupers, thus providing a basis for effective control and prevention of this disease.     

Effects of fishmeal replacement with cottonseed protein concentrate on growth,digestive proteinase,intestinal morphology and microflora in pearl gentian grouper (♀Epinephelus fuscoguttatus × ♂Epinephelus lanceolatu)

A 56‐day feeding trial was conducted to study the effects of cottonseed protein concentrate (CPC) as fishmeal (FM) replacement on growth, digestive proteinase, intestinal morphology and microflora in pearl gentian grouper (♀Epinephelus fuscoguttatus × ♂Epinephelus lanceolatu). FM of 0% (FM), 12.00% (R12), 24.00% (R24), 36.00% (R36), 48.00% (R48) and 60.00% (R60) was replaced with CPC, to prepare six diets and feed groupers. Weight gain (WG) and specific growth rate (SGR) were highest in the R24 group, and the lowest feed coefficient rate (FCR) was observed in this group compared with the other groups (p < .05). Pepsin activity of the R12 group and chymotrypsin activity of the R36 group reached significant maximum (p < .05). With increasing CPC replacement, the plica height and width and the lamina propria thickness of intestine decreased significantly (p < .05). 16S rDNA amplicon sequencing suggested dominant intestinal microflora of grouper subjected to different treatments was similar. At the phylum level, the abundance of Proteobacteria, Firmicutes, Verrucomicrobia, Bacteroidetes and Actinobacteria was high, while Photobacterium, Pseudoxanthomonas, Romboutsia, Akkermansia, Vibrio and Bifidobacterium were the dominant taxa at the genus level. The abundance of probiotics (Bifidobacterium) decreased, whereas the abundance of opportunistic pathogen (Proteobacteria, Actinobacteria and Vibrio) increased. These results suggested that CPC replacement affected growth, digestive proteinase activity, intestinal morphology and microflora structure in grouper. Under the conditions of this experiment, the fishmeal replacement with CPC (24% level) yielded the most satisfactory results, providing reference for sustainable aquaculture.     

The high prevalence of pathogenic Vibrio harveyi with multiple antibiotic resistance in scale drop and muscle necrosis disease of the hybrid grouper,Epinephelus fuscoguttatus (♀) × E. lanceolatus (♂), in China

Scale drop and muscle necrosis disease with high mortality widely occurred recently in the hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂), a crucial cultured marine fish species in China. In this study, 30 Harveyi clade isolates of 27 Vibrio harveyi strains were isolated from diseased hybrid groupers in the south‐east and north‐east coastal areas of China. A total of 22 V. harveyi strains were determined to be pathogenic, and most challenged fish died within 2 days of infection; surviving individuals exhibited scale drop and deep dermal lesions as naturally diseased fish. Although five typical virulence genes, including luxR, toxR_Vh, chiA, serine protease and vhh widely existed in V. harveyi, no obvious correlation was established between virulent strains and virulence genes harboured in them. Furthermore, multiple antibiotic resistance was widely exhibited in Harveyi clade strains, particularly for penicillins, polypeptides, lincomycins, acetylspiramycin, streptomycin, metronidazole and bacitracin. And the multiple antibiotic resistance indices were gradually decreased from southern to northern areas of China. This study demonstrated that the pathogenic V. harveyi with multiple antibiotic resistance is highly prevalent in hybrid grouper in China, which requires particular attention.     

Soy protein concentrate as an alternative in replacement of fish meal in the feeds of hybrid grouper,brown‐marbled grouper (Epinephelus fuscoguttatus) × giant grouper (E. lanceolatus) juvenile

Hybrid grouper juveniles (body weight, 6.1 ± 0.7 g) (brown‐marbled grouper, Epinephelus fuscoguttatus × giant grouper, E. lanceolatus) were fed with six isoproteic (50% crude protein) and isolipidic (12% crude lipid) feeds containing different levels of soy protein concentrate (SPC) in replacement of fish meal (SPC at 20%, 30%, 40%, 50% and 60% protein) and control feed (SPC0) for 6 weeks. Hybrid grouper juveniles were cultured in 100‐L fibreglass tank equipped with flow‐through water system and fed twice a day to apparent satiation level. The highest and lowest growth was recorded in fish fed SPC20 and SPC60 respectively. However, growth of SPC20 was not significantly higher than those fed SPC0, SPC30, SPC40 and SPC50 (p > .05). A decreasing growth trend was observed with the increasing level of SPC from feed SPC40 to SPC60. A noticeable better feed utilization was also observed in fish fed SPC0, SPC20, SPC30, SPC40 and SPC50 compared to fish fed SPC60 (p < .05). The fish condition factor, hepatosomatic index, viscerosomatic index and whole body proximate content of the fish were not affected by the graded levels of SPC. However, the body lipid content was significantly lower in fish fed SPC40 to SPC60 (p < .05). The apparent digestibility coefficient (ADC) of protein and lipid was significantly higher in fish fed SPC0 and SPC20 compared to other dietary treatments (p < .05). Based on the regression analysis on specific growth rate, the study suggests that the hybrid grouper grow best at 21.4% and can utilize up to 50% inclusion level of SPC in protein without significantly affect their growth and its body condition.     

Growth performance and disease resistance against Vibrio vulnificus infection of novel hybrid grouper (Epinephelus lanceolatus × Epinephelus fuscoguttatus)

Groupers are economically important for aquaculture in Thailand. A novel hybrid grouper (Epinephelus lanceolatus × Epinephelus fuscoguttatus) has been successful cross‐bred; therefore, the present work aimed to assess the hybrid's traits. The growth performance, strength and tolerance to a pathogenic bacterial infection of this hybrid were compared with its parent species, tiger grouper and giant grouper. The results of all measured growth parameters indicated that the hybrid strain grew fastest followed by giant and tiger grouper respectively. The expressions of the growth‐related genes, insulin‐like growth factor (IGF) I and II, were also analysed in fish muscle and liver which are the main target organs in fish growth regulation. Among tested species, similar expression patterns of IGF‐I and IGF‐II were detected in both organs. The levels of these genes in liver and muscle of hybrid and giant grouper were higher than those of tiger grouper comparable with the growth manner. After challenge with Vibrio vulnificus, the immunological parameters, clearance time of Vibrio in haemolymph and survival was measured to verify the fish immunity. Leucocyte number, lysozyme activity and the ability to eliminate the pathogen were very high in hybrid and giant grouper while these parameters were lower in tiger grouper. Correspondingly, the mortality rate of tiger grouper was higher than others and % survival at the end of observation time (15 days post challenge) was lowest in infected tiger grouper. Altogether, the results suggested that the hybrid grouper has desirable traits that will improve cultured grouper.     

Recent update on the prevalence of Vibrio species among cultured grouper in Peninsular Malaysia

Vibrio infections are common among marine fish and lead to serious problems in the aquaculture sector. This study reports a recent occurrence of Vibrio species (spp.) isolated from cultured groupers in Peninsular Malaysia using the gyrB and pyrH genes. A total of 147 Vibrio strains were successfully isolated from 77 (64%) groupers using culture method and subjected to gyrB and pyrH sequencing for species identification and confirmation. Results showed that 89% of Vibrio strains were identified and clustered to six groups of Vibrio spp., while 11% were not clustered to any Vibrio spp. using the gyrB sequences. Meanwhile, by analysis of the pyrH sequences all the 147 Vibrio strains (100%) were successfully identified and clustered into 11 groups of Vibrio spp., including the gyrB non‐identified strains. The pyrH gene provides a better resolution for identification of Vibrio spp. compared with the gyrB gene. Thus, the pyrH gene was more suitable for a rapid determination of Vibrio spp. distribution in Peninsular Malaysia. Using the pyrH gene, our study found higher prevalence of Vibrio vulnificus (33%), V. alginolyticus (24%) and V. parahaemolyticus (22%), followed by V. rotiferianus (5%), V. harveyi (3%), V. tubiashii (2%), V. campbellii (2%), V. ponticus (1%), V. diabolicus (1%), V. owensii (1%) and others Vibrio sp. (7%). Thus, the results of this study revealed that the occurrence of pathogenic vibrios among grouper fish is still high in Malaysian aquaculture. In addition, the pyrH gene was proved as a suitable marker for rapid identification of Vibrio species compared with the gyrB gene.     

The effects of different fishmeal level diets with or without phytase supplementation on growth performance,body composition,digestibility, immunological and biochemical parameters of juvenile hybrid sturgeon (Acipenser baeri Brandt ♀ × A. schrenckii Brandt ♂)

A 12‐week growth trial was conducted to evaluate the effect of low fishmeal and phytase replacing inorganic phosphorus on juvenile hybrid sturgeon (Acipenser baeri Brandt ♀ × A. schrenckii Brandt ♂). Four experimental diets were designed as FM100p, FM100, FM250p and FM250, which represented 10% and 25% fishmeal with or without phytase, respectively. The results showed that although growth parameters had no significant difference in all treatments, low fishmeal obviously decreased the culture cost of hybrid sturgeon. Lipid content of body composition was obviously increased in low fishmeal treatments (p < .05). The interaction of low fishmeal and phytase significantly enhanced apparent digestibility coefficients of crude protein (p < .05). Two‐way ANOVA analysis showed that input and output of phosphorus were significantly affected by fishmeal level, phytase and interaction of both factors (p < .05). For the immunological and hepatic function parameters, fishmeal level and phytase had significant effects on the activities of lysozyme, myeloperoxidase, malondialdehyde, alanine aminotransferase, lactate dehydrogenase and alkaline phosphatase (p < .05). The present study potentially justifies that a diet containing 10% fishmeal can be used for hybrid sturgeon. Moreover, replacement of monocalcium phosphate by phytase not only has no adverse effect on the growth of hybrid sturgeon, but also reduces phosphorus emission and protects aquaculture environment.     

Effects of alternative dietary oils on lipid metabolism and related gene expression in hybrid grouper (♀Epinephelus fuscoguttatus × ♂E. lanceolatu)

An 8‐week growth trial was conducted to evaluate effects of dietary oil sources on growth, enzymes activity and genes expression levels related to lipid metabolism of hybrid grouper (♀Epinephelus fuscoguttatus × ♂E. lanceolatu) juveniles. Seven iso‐lipid (97 g/kg of dry matter) and iso‐protein (503.5 g/kg of dry matter) experimental diets were formulated containing 50 g/kg fish oil (FO; acting as controls) or various vegetable oils (VOs): corn oil (CO), sunflower oil (SO), tea oil (TO), olive oil (OO), rice oil (RO) and mixed oil (MO; comprising equal amounts of these oils). Each diet was fed to triplicate groups of 40 fish for per repetition (15.09 ± 0.01 g) for 56 days. The results show that (a) alternative dietary oils had no significant effects on final weight compared with control group (p > .05); (b) compared with FO group, VOs significantly changed the contents of serum lipoproteins, cholesterol, triglycerides and the activity of liver lipid‐metabolizing enzymes (p < .05); (c) CO group had the least effect on the serum lipoproteins, triglycerides and cholesterol of grouper compared with control; the activity of liver lipid‐metabolizing enzymes in RO and control group was the closest; (d) the mRNA levels of Δ6 Fatty acid desaturase (Δ6Fad), hormone‐sensitive lipase (HSL) and lipoprotein lipase (LPL) were not significantly effected by lipid sources, but CO, TO, OO and MO significantly down‐regulated the expression of fatty acid synthetase (FAS) mRNA level in liver, while RO opposite (p < .05); (e) vegetable oil significantly up‐regulated peroxisome proliferator‐activated receptor α (PPARα) and peroxisome proliferator‐activated receptor β (PPARβ) mRNA levels, while TO and RO down‐regulated peroxisome proliferator‐activated receptor γ (PPARγ) mRNA levels (p < .05); and 6) MO significantly increased the mRNA levels of heart‐type fatty acid‐binding protein (H‐FABP) and adipocyte‐type fatty acid‐binding protein (A‐FABP) (p < .05), while other VOs had no effect on them (p > .05). In conclusion, dietary substitution of FO by VO in diet affected lipid metabolism of grouper, which may be regulated by PPARs.     

Antiviral activity of Illicium verum Hook. f. extracts against grouper iridovirus infection

Grouper iridovirus causes high mortality rates in cultured groupers, and effective treatment for grouper iridovirus infection is urgently required. Illicium verum Hook. f. is a well-known medicinal plant with a variety of biological activities. The aim of this study was to analyse the use of I. verum extracts to treat grouper iridovirus infection. The safe working concentration of each I. verum extract was identified both in vitro and in vivo as follows: I. verum aqueous extract (IVAE) ≤ 500 μg/ml; I. verum ethanol extract (IVEE) ≤ 250 μg/ml; shikimic acid (SKA) ≤ 250 μg/ml; trans-anethole (TAT) ≤ 800 μg/ml; 3,4-dihydroxybenzoic acid (DDBA) ≤ 400 μg/ml; and quercetin (QCE) ≤ 50 μg/ml. The inhibitory activity of each I. verum extract against grouper iridovirus infection was analysed using aptamer (Q2)-based fluorescent molecular probe (Q2-AFMP) and RT-qPCR. All of the I. verum extracts displayed dose-dependent antiviral activities against grouper iridovirus. Based on the achieved per cent inhibition, IVAE, IVEE, DDBA and QCE were associated with the greatest antiviral activity (all > 90%). Together, our results indicate that I. verum extracts have effective antiviral properties, making it an excellent potential source material for the development of effective treatment for grouper iridovirus infection.     

Effects of dietary inclusion of soybean meal and cholesterol on the growth,cholesterol status and metabolism of the giant grouper (Epinephelus lanceolatus)

The study evaluated effects of cholesterol supplementation in a diet with high soybean meal (SBM) on the growth and cholesterol metabolism of giant grouper (Epinephelus lanceolatus). All‐fish‐meal diet was used as control. The diet including SBM (replaced 50% of the fish meal protein, SBM diet) and the SBM diet supplemented with 10 g/kg cholesterol (SBM + cholesterol) were used as experimental diets. Three diets were each fed to triplicate groups of juvenile grouper (initial body weight: 12.39 ± 0.36 g) in a recirculating aquaculture system for 8 weeks. Grouper fed the control diet showed higher (p < .05) weight gain, feed intake, feed efficiency and protein efficiency ratio than the other two dietary treatments. Hepatic cholesterol concentrations and 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase gene expressions were higher in fish fed the control diet than fish fed the control diet and SBM + cholesterol diet. Hepatic cholesterol 7α‐hydroxylase gene expression was higher in fish fed the SBM + cholesterol diet than that in fish fed the control diet. Results indicate that giant grouper on a diet low in cholesterol can regulate cholesterol synthesis, suggesting that the reduced dietary cholesterol intake in the fish fed diet containing SBM is sufficiently compensated by increased cholesterol synthesis.     

云纹石斑鱼、鞍带石斑鱼及杂交“云龙斑”肌肉营养成分分析及品质评价

对云纹石斑鱼、鞍带石斑鱼及杂交"云龙斑"肌肉营养成分进行分析和品质评价,为其种质营养学鉴定及配合饲料的研制提供基础数据资料。采用常规方法,依据国家标准,对3种石斑鱼肌肉中的水分、蛋白、粗脂肪、灰分含量以及氨基酸、脂肪酸和10种矿物元素的组成与含量进行了测定和评价。云纹石斑鱼、鞍带石斑鱼及杂交"云龙斑"石斑鱼肌肉中粗蛋白含量分别为19.6%、18.4%和19.4%;3种石斑鱼氨基酸总量、必需氨基酸总量、必需氨基酸指数、鲜味氨基酸总量分别为(15.54%、14.72%、15.99%)、(6.86%、6.41%、7.04%)、(75.19、74.93、77.96)和(5.57%、5.3%、5.73%)。3种石斑鱼肌肉中粗脂肪含量分别为4.2%、10.1%和4.5%,脂肪酸种类丰富,并且含有大量的多不饱和脂肪酸,其比例符合理想脂肪酸的标准。3种石斑鱼肌肉中矿物元素种类丰富,5种常量元素K、Ca、Na、Mg、P和5种微量元素Fe、Mn、Cu、Zn、Se均有检出,其中杂交"云龙斑"的Na、K、Cu含量略高,另外几种矿物元素含量居于父母本之间。研究表明,云纹石斑鱼、鞍带石斑鱼及杂交"云龙斑"氨基酸种类齐全、比例适宜,人体必需氨基酸含量较高,是优质的蛋白源,同时含有大量的不饱和脂肪酸,且矿物元素种类齐全,是具有较高食用价值的优良海水养殖品种。其中杂交"云龙斑"在一些营养指标中明显高于其父母本,显现出一定的杂交优势。     

Effects of dietary supplementation with coconut oil on the growth,fatty acid profiles and some lipid metabolism relative gene expressions of orange‐spotted grouper Epinephelus coioides

This study investigated the effects of coconut oil as a dietary supplement on the growth, lipid metabolism and related gene expressions of juvenile orange‐spotted grouper Epinephelus coioides. Coconut oil at concentrations of 0, 10, 30 and 50 g/kg was used to replace dietary lipids in a basal diet containing 150 g/kg lipids. The four experimental diets were, respectively, fed to triplicate groups of juvenile groupers (initial weight: 8.53 ± 0.13 g) in a recirculating system for 8 weeks. Fish fed the diet containing 50 g/kg coconut oil exhibited lower (p < .05) weight gain than did fish fed the diet containing 30 g/kg coconut oil; however, no significant differences in weight gain were observed between fish fed diets containing 0 and 10 g/kg coconut oil. Hepatic carnitine palmitoyltransferase‐1, fatty acid synthase, fatty acid elongase, fatty acid desaturase and peroxisome proliferator‐activated receptor gamma gene expressions were all the highest in fish fed the diet containing 10 g/kg coconut oil. Fish fed the coconut oil‐free basal diet demonstrated upregulated gene expression of neuropeptide Y. The results suggest that dietary supplementation with 10 g/kg coconut oil exerted beneficial effects on lipid metabolism by E. coioides.     

Grouper aquaculture: Asian success and Mediterranean trials

• 1. Grouper are of considerable economic value in tropical and subtropical regions and most particularly in south-east Asia. Moreover, it is in Asia, where this fish is highly prized, that the grouper farming began in the 1980s.
• 2. The organoleptic qualities of the groupers are not the only characteristics to be appreciated; in culture, their robustness in heavily populated conditions, as well as their rapid growth at elevated temperatures makes them a good species for aquaculture. However, it is, above all, market demand that is outstripping the supply of fish, which is motivating the expansion of grouper aquaculture.
• 3. At the present time, aquaculture is unable to satisfy demand. The chronic shortage of broodstock means that the alevins, destined for aquaculture, are taken mainly from natural habitats with the aid of fish traps, nets and cyanide, thus contributing to the destruction of south-east Asian reefs. Nevertheless, research is progressing and the number of species whose reproduction is totally controlled is increasing. It is in Taiwan that grouper aquaculture is at its most advanced with a total of five species being raised.
• 4. Aquaculture is divided into four stages, carried out by different farmers, allowing a more rapid turnover for each farmer and therefore a better financial return. On the other hand, in the Mediterranean at present time, there are no completely controlled grouper captive-breeding programmes. The first attempts at controlled breeding have been carried out since 1995 in Italy, Croatia and Greece. However, farming groupers in captivity, and in particular the control of their reproduction, poses numerous problems.

Copyright © 2007 John Wiley & Sons, Ltd.     

Use of Onion (Allium cepa) and Garlic (Allium sativum) Wastes for the Prevention of Fungal Disease (Saprolegnia parasitica) on Eggs of Rainbow Trout (Oncorhynchus mykiss)

This study was designed to prevent fungal disease (Saprolegnia parasitica) that occurs on rainbow trout eggs (Oncorhynchus mykiss) by using wastes of onion (Allium cepa) and garlic (Allium sativum) plants. For this purpose, fertilized rainbow trout eggs were exposed to garlic skin, garlic stem and onion skin aqueous methanolic extracts by bathing in concentrations of 0.4, 0.8, 1.6 and 3.2 g/L, whereas the control group was left untreated. The larvae in all groups were monitored until they become free-swimming larvae, the number of eggs died due to fungus was recorded, and the data obtained from experimental groups and control group were compared. As a result, it was determined that onion skin had no effect on the number of eggs that died due to fungus (p > .05), while garlic skin and garlic stem extracts significantly reduced the number of fungal infestations without affecting the number of live larvae or the embryological development of the eggs (p < .05). In light of these data, we conclude that garlic skin (0.4, 0.8, and 1.6 g/L) and garlic stem (0.8 and 1.6 g/L) aqueous methanolic extracts are effective at preventing Saprolegnia parasitica infestation on rainbow trout eggs and may be used in aquaculture.     

Ecological and histopathological aspects of Didymodiclinus sp. (Trematoda: Didymozoidae) parasite of the dusky grouper,Epinephelus marginatus (Osteichthyes: Serranidae), from the western Mediterranean Sea

The dusky grouper Epinephelus marginatus (Lowe) is an ecologically and commercially important fish species of the Atlantic and Mediterranean coastal rocky habitats. Despite records of didymozoid infections in several grouper species, the identification and pathogenesis of these parasites in E. marginatus are lacking. The aim of this study is to characterize the didymozoids of E. marginatus, particularly their mechanisms of infection and histopathological features. Dusky groupers (n = 205) were caught off Majorca Island (western Mediterranean Sea) and examined for parasites. Of the fish sampled, 45% were infected with white and yellow didymozoid capsules and brown nodules, found on the gills and pseudobranchs. Parasite abundance had a strong positive relationship with the fish length; only fish larger than 20 cm were infected, suggesting infection via consumption of an intermediate host, for which E. marginatus size was a limiting factor. The capsules contained two convoluted viable adult trematodes, identified as Didymodiclinus sp., in close contact with host capillary vessels, with no evidence of the tissue inflammatory response. Conversely, nodules containing degraded parasites were surrounded by an intense inflammatory infiltrate. The findings suggest that Didymodiclinus sp. have the potential to evade the host's immune system by inhibiting the inflammatory response.     

Can barber goby Elacatinus figaro control Neobedenia melleni infections on dusky grouper Epinephelus marginatus?

The interaction between cultured barber goby Elacatinus figaro (cleaner fish) and the dusky grouper Epinephelus marginatus, and the efficiency of the cleaner fish in removing ectoparasites were evaluated. When the interaction between these two species was observed, cleaner fish showed a preference for the largest groupers. In a second trial, treatments: TWC (Control) – two groupers without a cleaner fish, T1C – two groupers with one cleaner fish, T3C – two groupers with three cleaner fish and T6C – two groupers with six cleaner fish were tested in four replicates. After 8 days, monogeneans were removed and identified as Neobenedenia melleni. The highest mean abundance of parasites was found on the groupers in the TWC group (37 parasites per host) and the lowest on those in the T6C group (4.1 parasites per host). By increasing the number of cleaner fish, a higher cleaning efficiency was obtained, as observed in T6C, where almost 90% of the parasites were removed. Possibly, this removal would have been complete if the number of cleaners had not been reduced in the treatments due to the mortalities observed. This study demonstrates the possibility of using gobies to remove monogeneans and in improving grouper health.