Effect of guava Psidium guajava (L.) aqueous extract diet on growth performance,intestinal morphology,immune response and survival of Oreochromis niloticus challenged with Aeromonas hydrophila

An 84‐day feeding trial was carried out to evaluate the effects of aqueous Psidium guajava leaf extract (PGE) on growth, intestinal morphology, physiology, immune response and susceptibility of Oreochromis niloticus to Aeromonas hydrophila. Diets containing 0% (P0), 0.25% (P1), 0.50% (P2), 0.75% (P3) and 1.00% (P4) PGE were fed to triplicate groups of fish (mean weight; 1.32 ± 0.04 g) for 84 days. After the 84‐day feeding trial, test fish were injected with pathogenic A. hydrophila and then fed for 14 days. More feed were consumed in groups of fish fed PGE‐treated diets and resulted in significantly higher weight gain and feed intake. Incidentally, there was an increase in the calculated area of absorption of fish fed PGE diets, as accounted for by marginally higher villi length and width. Antioxidant and immune response were improved with PGE inclusion in diets as total protein, superoxide dismutase, glutathione peroxidase and glutathione S‐transferase significantly increased (p < 0.05) in fish fed PGE diets. Results of the challenge test with A. hydrophila revealed that the highest mortality (100%) was recorded in P0. This study revealed that inclusion of P. guajava extract in the diet of O. niloticus improved growth, nutrient utilization, immune system and survival of O. niloticus fingerlings.     

Performance of Oreochromis niloticus in recirculating aquaculture systems at different levels of daily protein intake

Recirculating aquaculture systems (RASs) are an efficient method for developing sustainable aquaculture and the development of a feed strategy for RASs could be a key factor for the success of culturing Oreochromis niloticus. The daily protein intake (DPI) mathematical function related to body weight (BW) was designed (DPI = ?3.818 ln (BW) + 30.158) named DPI 1.0 = 100%. Three feeding rates were established, DPI 1.4, 1.2 and 1.0, and were implemented in an RAS production cycle (100 fish/m³) for 34 weeks. The final growth measurements were 908.0 ± 57.9 g (DPI 1.4), 887.0 ± 113.5 g (DPI 1.2) and 702.2 ± 38.1 g (DPI 1.0) (p < .05), with feed conversion rates (FCRs) of 2.3, 1.9 and 1.9, respectively; the survival was ≥ 98.3% (p > .05) for all treatments. DPI levels conditioned the growth of the fish and the proportion of lipid: protein in the bodies of O. niloticus but not in the fillets. The mathematical function DPI 1.2 = 120% (DPI = ?4.582 ln (BW) + 36.19) used in the RAS at the 40% protein level improved fish growth, the protein efficiency ratio and FCRs. The DPI values were estimated for 49 different cases and feeding strategies showed lower values than most of the different DPI mathematical functions used and cited in this work. Therefore, DPI metric is a good tool that provides the specific requirements of protein for O. niloticus growth, which can enable farmers to ration feed and improve energy use (kWh / fish) and facilitate the integration of RAS with horticulture.     

Earthworm,Eisenia fetida,bedding meal as potential cheap fishmeal replacement ingredient for semi‐intensive farming of Nile Tilapia,Oreochromis niloticus

The study amalgamated earthworm and agro‐industrial wastes through vermicomposting and then evaluated the potential of the bedding (mixture of Eisenia fetida and vermicompost) to replace fishmeal in semi‐intensive farming of Oreochromis niloticus. The bedding was used to substitute fishmeal at inclusion rates of 100, 60, 30 and 0% (D100, D60, D30 and control D0). In triplicates, the four homogeneous diets were fed to quadruplicate groups of 30 g O. niloticus for 112 days. There was no significant difference (p > .05) in mortalities, average length gain and FCR among all tests. Nevertheless, diet D0 had significantly (p < .05) superior amino acid profile, low fibre content and fish carcass crude protein (63.2 ± 0.72% dry matter). Subsequently, D30 and D0 produced fish with significantly higher (p < .05) mean weight gain (256.03 ± 0.4 g) and biomass (369,136 g) respectively. On to the contrary, diet D100 had significantly higher (p < .05) crude lipids content (9.4 ± 0.6% dry matter), economic returns and profit index than the control diet due to the comparatively low cost of producing the earthworm bedding. This simple biotechnology can commercially be upscaled to sustainably produce cheap and nutritious fish feed capable of increasing yields and maximizing profits.     

Prevalence and effects of the cymothoid isopod (Ceratothoa oestroides,Risso 1816) on cultured meagre (Argyrosomus regius,Asso 1801) in the Eastern Adriatic Sea

Meagre (Argirosomus regius) is a relatively new farmed fish species in Croatian mariculture. This study is the first attempt to determine the prevalence of cymothoid isopod parasite Ceratothoa oestroides and its effects on cultured meagre in the Mediterranean area. When analysing growth of meagre at two sites in the central part of the Eastern Adriatic Sea, the greatest difference in total fish length was recorded in March 2016, when fish with parasites were 33% smaller than fish without parasites (24.47 ± 3.29 vs. 16.28 ± 1.01 cm; p < .001). The largest difference in mass was also recorded during sampling in March, when parasitized fish was 74% lighter than the unparasitized fish (171.75 ± 72.96 vs. 44.77 ± 6.57 g; p < .001). The presence of the cymothoid isopod in the buccal cavity was observed, and caused fish deformation throughout the entire duration of the study. Also, female parasites with pulli II in marsupium were found. Based on the determined prevalence and effects on production and health status, monitoring of the interaction between C. oestroides and meagre should be considered in future management plans for cultivation of this promising species for Mediterranean aquaculture.     

Nematode parasites infecting the starry batfish Halieutaea stellata (Vahl) (Lophiiformes: Ogcocephalidae) from the East and South China Sea

The starry batfish Halieutaea stellata (Vahl) is a small, benthic fish found in Indo‐West Pacific Oceans. However, our present knowledge of the helminth parasites of this fish is still fragmentary. In this study, a total of 29 fish collected from the East and South China Sea were examined to determine the prevalence, intensity and species composition of helminth parasites in H. stellata. Using morphological and molecular approaches, four species of nematodes were found parasitic in this fish host, including the adults and fourth‐stage larvae of Raphidascaroides nipponensis Yamaguti 1941; adults and third‐stage larvae of Raphidascaris lophii (Wu 1949), third‐ and fourth‐stage larvae of Hysterothylacium larval type IV‐A of Shamsi, Gasser & Beveridge 2013 and third‐stage larvae of Hysterothylacium amoyense (Hsü 1993). Halieutaea stellata represents a new host record for the three last‐named nematodes. Raphidascaroides nipponensis with the highest prevalence (82.5%) and intensity (mean = 13.5) of infection was considered as the dominant parasite species in H. stellata. The detailed morphology of the different developmental stages of the four nematode species was studied using light and scanning electron microscopy. All nematode species were also genetically characterized by sequencing and analysing the internal transcribed spacer (ITS) of the ribosomal DNA. This study provides further data on the occurrence of nematode parasites in H. stellata and also contributes to facilitate an accurate and rapid diagnosis of the infection by these little‐known nematodes.     

Detection of tilapia lake virus (TiLV) infection by PCR in farmed and wild Nile tilapia (Oreochromis niloticus) from Lake Victoria

Tilapia lake virus disease (TiLVD) has emerged to be an important viral disease of farmed Nile tilapia (Oreochromis niloticus) having the potential to impede expansion of aquaculture production. There is a need for rapid diagnostic tools to identify infected fish to limit the spread in individual farms. We report the first detection of TiLV infection by PCR in farmed and wild Nile tilapia from Lake Victoria. There was no difference in prevalence between farmed and wild fish samples (p = .65), and of the 442 samples examined from 191 fish, 28 were positive for TiLV by PCR. In terms of tissue distribution, the head kidney (7.69%, N = 65) and spleen (10.99%, N = 191), samples had the highest prevalence (p < .0028) followed by heart samples (3.45%, N = 29). Conversely, the prevalence was low in the liver (0.71%, N = 140) and absent in brain samples (0.0%, N = 17), which have previously been shown to be target organs during acute infections. Phylogenetic analysis showed homology between our sequences and those from recent outbreaks in Israel and Thailand. Given that these findings were based on nucleic acid detection by PCR, future studies should seek to isolate the virus from fish in Lake Victoria and show its ability to cause disease and virulence in susceptible fish.     

The urogenital papillae as an indicator of sexual maturity in male African catfish,Clarias gariepinus, (Burchell, 1822)

The present study investigated a non‐invasive method based on macroscopic morphological features of male Clarias gariepinus for the assessment of sexual maturity stage. As African catfish cannot be stripped for semen as in most other fish species; they have to be killed to obtain semen from the testes. This method could be used to ensure that males used to obtain sperm are sexually mature to increase the concentration and quality of mature sperm for egg fertilization in the production of C. gariepinus. Morphological features including the length and width of the urogenital papillae (UGP), an external secondary sexual characteristic of C. gariepinus (n = 89) were measured and calculated as a percentage of total body length. The urogenital papillae length (L_UGP) and width (W_UGP) index of each fish were used and compared with the macroscopic and microscopic maturity stage of the testes. The results revealed that although positive, a significant correlation does not exist between the L_UGP or W_UGP and maturity stage of the testes. Therefore, although this procedure would be valuable as an initial non‐invasive assessment of maturity, it does not provide an accurate or reliable indication of the maturity stage of the fish.     

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.     

Effect of Arthrospira supplementation on Oreochromis niloticus gut microbiota and flesh quality

Supplementing fish feed with Arthrospira platensis has several beneficial effects, such as improved growth, survival and feed conversion, but little is known about how it affects gut microbiota or flesh quality. In this trial, feed with 3% A. platensis was given to Oreochromis niloticus (SPR, spirulina supplemented) over a period of 50 days and their gut microbiota and flesh quality were compared with controls (CTRs, no supplementation). Average daily growth was higher in SPR fish (p < 0.05). Regarding microbiota, a total of 5,470,272 readings (individuals) were obtained from 16 gut samples (n = 8 CTR, n = 8 SPR), with a gamma diversity of 1,563 species but no significant differences were found between treatments. Among the high inter‐individual variability found, some individual microbiota were dominated by Proteobacteria (36.6%) and Fusobacteria (32.9%) and others more by Actinobacteria (37%). Regarding flesh quality, significant differences were observed in flesh colour post‐mortem (a*, b* and chroma) (p < 0.05), with higher reflectance in the range of 470–500 and 620–670 nm in SPR tilapia, which corresponds with the reflectance range of β‐carotene and phycocyanin. However, rigor mortis was resolved earlier in SPR fish (p < 0.05), which suggests a shorter shelf life. Adding 3% of A. platensis in O. niloticus diet can improve growth and flesh colour and does not appear to modify intestinal microbiota.     

Practical diets with essential oils of plants activate the complement system and alter the intestinal morphology of Nile tilapia

The effect of the essential oils (EOs) of peppermint, Mentha piperita L., and tea tree, Melaleuca alternifolia (Maiden & Betche) Cheel, on the haematological, biochemical, and immunological parameters and intestinal morphology of Nile tilapia, Oreochromis niloticus L., was evaluated. Fish (58.09 ± 5.87 g) were fed 100 mg/kg and 250 mg/kg of each EO and sampled on days 7, 14, 30 and 60 after starting supplementation. The haematological and biochemical parameters were not altered by the supplementation of EOs compared to the control (p > .05). With regard to the immunological parameters, the activation of the complement system of fish fed 250 mg/kg peppermint and 100 mg/kg and 250 mg/kg tea tree EOs were significantly higher compared to the control after 60 days of feeding (p < .05). The complement system plays an essential role in innate immunity and contributes significantly to the acquired immune response; thus, its activation through supplementation with EOs is promising for the formulation of nutritional additives in aquaculture. Regarding intestinal morphology, fish fed 250 mg/kg tea tree EO presented higher villus size compared to all other groups (p < .05), which represents a healthier gut. These fish present a larger intestinal surface, which can result in better absorption and utilization of the nutrients. Based on the responses found in this study, both EOs were considered promising for the formulation of feed additives for Nile tilapia.     

Formulated diet alters gut microbiota compositions in marine fish Nibea coibor and Nibea diacanthus

Nibea, an economical marine fish, is generally fed on trash fish (the low‐value fish), which can cause high feed costs and waste pollutions in high‐density aquaculture. To assess the effect of formulated diet on the gut microbiota in Nibea coibor and Nibea diacanthus, denaturing gradient gel electrophoresis, clone libraries analysis and Illumina sequencing of the 16S ribosomal RNA gene were used in this study. Two Nibea fishes were both dominated by Gammaproteobacteria (especially Photobacterium) and shared a set of gut microbiota, including Bacilli, Mollicutes, Alphaproteobacteria and Fusobacteriia. Statistical analyses revealed that formulated diet led to lower feed conversion ratio (p < 0.001), lower abundance of Vibrio (p = 0.040) and infectious diseases pathways (p = 0.001), higher abundance of polysaccharide‐degrading bacterium Cellvibrio (p = 0.006) in two Nibea species, with higher weight gain rate (p = 0.023) and microbial diversity (Shannon, p = 0.049 and Simpson, p = 0.044) and more carbohydrate metabolism (p = 0.020) observed in N. coibor. The distribution and correlation network of 17 potential short‐chain fatty acid producing bacteria were obtained and visualized in all treatment groups. The results reveal that formulated diet has beneficial effects on the gut microbial ecology in two Nibea fishes, which suggests the possibility of replacing trash fish diet with formulated diet in Nibea aquaculture.     

Use of neem oil (Azadirachta indica) to control caligid copepod infestation on Asian seabass (Lates calcarifer)

Asian seabass (Lates calcarifer) culture in marine cages have been developed rapidly in Malaysia. The high intensity of culture might facilitate several disease infection including caligid infestation. This study was conducted to evaluate the effect of neem oil (Azadirachta indica) against caligid parasites on seabass. The prevalence and clinical pathogen of Caligus infection has been described through a sampling of 150 fish from floating cage at different cultured periods. Acute toxic tests with different concentrations of neem oil were carried out on the parasite and fish host. Results revealed that the 96‐hr median lethal concentration (LC₅₀) of Caligus and fish (10.3 ± 2.5 g of body weight) were 2 and 20 ppm respectively. The in vivo test indicated that 10 ppm of neem oil could result in 100% antiparasitic efficacy within 96 hr. These results therapeutically demonstrated the efficacy of neem oil in caligid control.     

Effects of dietary probiotic Lactobacillus plantarum and whey protein concentrate on the productive parameters,immunity response and susceptibility of Nile tilapia,Oreochromis niloticus (L.), to Aeromonas sobria infection

Immunostimulatory feed supplements have an increasingly interest in aquaculture management. Generally, an individual supplement was used in fish diets but it is expected that the use of multi‐supplements may show synergistic enhancements in fish performance, health, and immunity. Therefore, the present investigation was carried out to evaluate the use of dietary probiotic Lactobacillus plantarum and whey protein concentrate (WPC) in practical diets for Nile tilapia, Oreochromis niloticus. Hence, probiotic L. plantarum, WPC and their mixture were incorporated into a basal fish diet (300 g/kg crude protein) as follows: T1 = a basal control diet, T2 = a basal diet containing L. plantarum, T3 = a basal diet containing 1.0 g WCP/kg diet and T4, T5 or T6 = basal diets containing probiotic L. plantarum + 1.0, 2.0 or 3.0 g WCP/kg diet, respectively. Fish (15.2 ± 0.6 g) were fed on one of the tested diets up to apparent satiation twice a day for 60 days. After that, fish were intraperitoneally injected with pathogenic bacteria Aeromonas sobria and fish mortality was observed for 10 days postchallenge. Fish growth and feed intake were significantly improved by dietary probiotic L. plantarum (T2) and/or WPC (T3) over the control group (T1), and highest fish performance was observed in T5–T6 fish groups. Similarly, highest values of haematocrit, glucose, total proteins, albumin, and globulin were significantly observed in T5–T6 fish groups. Likewise, fish fed dietary probiotic L. plantarum (T2), WPC (T3), and their mixture (T4–T6) showed antioxidants and immune‐stimulating activities better than the control group. Fish fed the control diet were more susceptible to A. sobria infection showing highest fish mortality (75.0%). Meanwhile, dietary probiotic L. plantarum (T2), WPC (T3), and their mixture (T4–T6) enhanced significantly the fish resistance to A. sobria infection resulting in maximum values of relative percent of fish survival (73.3%–80.0%) in T5–T6 groups. The present investigation recommended the use of probiotic L. plantarum with 2.0 g WPC/kg diet to improve the growth, antioxidant, immunity responses and tolerance of Nile tilapia to A. sobria infection.     

Zingiber officinale and Glycyrrhiza glabra,individually or in combination,reduce heavy metal accumulation and improve growth performance and immune status in Nile tilapia,Oreochromis niloticus

This study evaluated the impact of dietary ginger and liquorice supplementation on growth performance, physiological and histopathological profiles and heavy metal accumulation in Nile tilapia fingerlings. Fish (n = 1,800, 17.5 ± 0.11 g BW) were randomly distributed into four treatment groups in triplicates and received no supplementation (control group), 5 ml aqueous ginger extract/kg feed (ginger group), 4 ml aqueous liquorice extract/kg feed (liquorice group) or 2.5 ml ginger plus 2 ml liquorice aqueous extracts/kg feed (mix group). The ginger‐liquorice mix supply improved the growth performance and feed efficiency (p < .05), increased the haematocrit and haemoglobin (p < .05), leucocytes (p = .108), neutrophils (p = .054), serum total protein (p < .05), albumin (p = .011) and globulin (p = .094) but decreased (p < .05) the blood urea nitrogen and creatinine than feeding liquorice or ginger lonely compared to the control. Heavy metal loads in pond water induced lamellar telangiectasis of gills and necrosis with sloughing of intestinal villi tips. These detrimental effects were alleviated, and the intestinal villus length (p = .041) and crypt depth (p = .069) were increased with liquorice supply. In all treatment groups, heavy metal contents in fish flesh were lower compared to the control. Thus, using ginger and/or liquorice aqueous extracts can decrease heavy metal accumulation in the fish flesh and exert positive effects on growth performance, metabolic profile and the intestinal and gill morphology of Nile tilapia.     

Beneficial effects on growth,haematic indicators,immune status,antioxidant function and gut health in Juvenile Nile tilapia (Oreochromis niloticus) by dietary administration of a multi‐strain probiotic

The present study was conducted to administer a commercial multi‐strain probiotic (MP), Yilibao (Bacillus velezensis:Bacillus cereus:Lactobacillus casei = 2:2:1), in Nile tilapia, Oreochromis niloticus. In terms of aerobic Bacillus spp. counts, we produced five diets containing 0, 0.34, 1.68, 3.36 and 6.72 g/kg of MP dry product (Control, T1, T2, T3, T4). Seven hundred and fifty tilapia juveniles (13.26 ± 0.01 g) distributed into 25 tanks in five replications were fed their diet for eight weeks. Results showed that fish fed T3 and T4 diets displayed significantly higher final body weight and weight gain (p < .05). Further, all MP‐treated fish exhibited remarkably decreased plasma lipid profiles (cholesterol, triglycerides) (p < .05) and fish fed T3 and T4 diets displayed significantly higher plasma myeloperoxidase activity and complement C3 content (p < .05). Additionally, fish fed T3 and T4 diets exhibited significantly promoted total antioxidant capacity, glutathione peroxidase activity, glutathione level and declined malondialdehyde content in the plasma and/or liver (p < .05). Moreover, significantly elevated chymotrypsin activity, villus height and intraepithelial lymphocytes counts were found in fish fed T3 and T4 diets (p < .05). As fish fed T3 and T4 diets surpassed the other treatments in growth, immune–antioxidative status and gut health, the recommended dose for tilapia is 3.36–6.72 g/kg.     

Effects of alternative dietary oils on immune response,expression of immune‐related genes and disease resistance in juvenile Nile tilapia,Oreochromis niloticus

The expression of immune‐related genes and immune responses to Aeromonas hydrophila were investigated on Oreochromis niloticus (6.07 ± 0.07 g), by feeding them six different diets for 8 weeks to apparent satiation. Diets contained fish oil (60g/kg FO), virgin coconut oil (60g/kg VCO) and corn oil (60g/kg CO) as sole lipids or blends of 30g/kg FO + 30g/kg VCO (3FVCO), 30g/kg FO + 30g/kg CO (3FCO) and 30g/kg VCO + 30g/kg CO (VO). Fish fed 3FCO recorded higher final weight, percentage weight gain (%WG) and specific growth rate (%SGR) but not significantly higher than all other groups. Triglyceride was higher in fish fed 3FCO than 3FVCO and CO (p ? 0.05), whereas total immunoglobulin (TIg) was not significant (p ? 0.05) between groups. Lysozyme activity was significantly higher in fish fed diet CO while groups FO, 3FCO and VCO recorded the least activities (p ? 0.05). Although alternative complement activity (ACH₅₀), complement proteins (C3 and C4), was not influenced, antibody titre production was significantly higher in fish fed diet 3FCO and lower in group CO. mRNA expression of IL‐1β was significantly upregulated in fish fed VO while the expression of C‐type lysozyme and TGF‐β was not significantly influenced across treatments, although group fed FO recorded higher expression levels, respectively. Lower mortalities of fish were recorded in groups fed 3FCO and VO after 14 days postchallenge with A. hydrophila disease indicating the enhancing effects of vegetable oils to boost immune response and resistance to disease. The study concludes that alternative lipid sources with high polyunsaturated fatty acids (PUFAs‐ALA and LA) (CO) and blend of saturated fatty acids (SFA)(VCO) can partially and or exclusively replace FO with an improved effect on tilapia and resistance to A. hydrophila in tilapia.     

Prevalence of Clonorchis sinensis infection in fish in South-East Asia: A systematic review and meta-analysis

Clonorchis sinensis, an important fish-borne zoonotic trematode, is widely distributed in South-East Asia, especially in China. Infections from human and animal reservoir hosts occur due to the consumption of raw or undercooked fish with C. sinensis metacercariae. This study aimed to evaluate the prevalence of C. sinensis metacercariae in fish in South-East Asia via systematic review and meta-analysis. We searched PubMed, ScienceDirect, China National Knowledge Infrastructure, Wanfang and Chongqing VIP databases for studies published between 1976 and 2020 that are related to the prevalence of C. sinensis metacercariae in fish. Studies were screened with keywords based on inclusion and exclusion criteria. Seventy-one eligible articles were identified, covering three countries: China, Korea and Vietnam. The pooled prevalence of C. sinensis metacercariae in fish from South-East Asia was 30.5%, with 35.1% in China, 29.7% in Korea and 8.4% in Vietnam. In subgroup analyses of climate, season, water source and publication date, the highest prevalence was identified in the Dwb climate type (43.3%), summer (70.2%), river (34.5%) and pre-2001 publications (38.9%), respectively. In comparison, the lowest prevalence was found in the Dfa climate type (14.5%), winter (19.5%), lake (8.0%) and post-2001 publications (23.8%). Meta-regression results indicated that country (p = .009), the published time (p = .035) and water source subgroups (p = .003) may be the source of heterogeneity. Overall, our study indicates that a high prevalence of C. sinensis infections occurs in fish in China, Korea and Vietnam, illuminating a significant public health concern in these countries.