Neem leaf powder (Azadirachta indica) mitigates oxidative stress and pathological alterations triggered by lead toxicity in Nile tilapia (Oreochromis niloticus).

This study investigated the clinical and pathological symptoms of waterborne lead toxicity in wild Nile tilapia collected from a lead-contaminated area (the Mariotteya Canal: Pb = 0.6 ± 0.21 mg L-1) and a farmed fish after 2 weeks of experimental exposure to lead acetate (5-10 mg L-1) in addition to evaluating the efficacy of neem leaf powder (NLP) treatment in mitigating symptoms of lead toxicity. A total of 150 fish (20 ± 2 g) were alienated into five groups (30 fish/group with three replicates). G1 was assigned as a negative control without any treatments. Groups (2-5) were exposed to lead acetate for 2 weeks at a concentration of 5 mg L-1 (G2 and G3) or 10 mg L-1 (G4 and G5). During the lead exposure period, all groups were reared under the same conditions, while G3 and G5 were treated with 1 g L-1 NLP. Lead toxicity induced DNA fragmentation and lipid peroxidation and decreased the level of glutathione and expression of heme synthesis enzyme delta aminolaevulinic acid dehydratase (ALA-D) in wild tilapia, G2, and G4. NLP could alleviate the oxidative stress stimulated by lead in G3 and showed an insignificant effect in G5. The pathological findings, including epithelial hyperplasia in the gills, edema in the gills and muscles, degeneration and necrosis in the liver and muscle, and leukocytic infiltration in all organs, were directly correlated with lead concentration. Thus, the aqueous application of NLP at 1 g L-1 reduced oxidative stress and lowered the pathological alterations induced by lead toxicity.

A First Report of Molecular Typing, Virulence Traits, and Phenotypic and Genotypic Resistance Patterns of Newly Emerging XDR and MDR Aeromonas veronii in Mugil seheli.

Aeromonas veronii is associated with substantial economic losses in the fish industry and with food-borne illness in humans. This study aimed to determine the prevalence, antibiogram profiles, sequence analysis, virulence and antimicrobial resistance genes, and pathogenicity of A. veronii recovered from Mugil seheli. A total of 80 fish were randomly gathered from various private farms in Suez Province, Egypt. Subsequently, samples were subjected to clinical, post-mortem, and bacteriological examinations. The retrieved isolates were tested for sequence analysis, antibiogram profile, pathogenicity, and PCR detection of virulence and resistance genes. The prevalence of A. veronii in the examined M. seheli was 22.5 % (18/80). The phylogenetic analyses revealed that the tested A. veronii strains shared high genetic similarity with other A. veronii strains from India, UK, and China. Using PCR it was revealed that the retrieved A. veronii isolates harbored the aerA, alt, ser, ompAII, act, ahp, and nuc virulence genes with prevalence of 100%, 82.9%, 61.7%, 55.3%, 44.7%, 36.17%, and 29.8%, respectively. Our findings revealed that 29.8% (14/47) of the retrieved A. veronii strains were XDR to nine antimicrobial classes and carried blaTEM, blaCTX-M, blaSHV,tetA, aadA1, and sul1 resistance genes. Likewise, 19.1% (9/47) of the obtained A. veronii strains were MDR to eight classes and possessed blaTEM, blaCTX-M, blaSHV,tetA, aadA1, and sul1 genes. The pathogenicity testing indicated that the mortality rates positively correlated with the prevalence of virulence-determinant genes. To our knowledge, this is the first report to reveal the occurrence of XDR and MDR A. veronii in M. seheli, an emergence that represents a risk to public health. Emerging XDR and MDR A. veronii in M. seheli frequently harbored aerA, alt, ser, ompAII, and act virulence genes, and blaTEM, sul1, tetA, blaCTX-M, blaSHV, and aadA1 resistance genes.

Analysis of the Productivity, Immunity, and Health Performance of Nile Tilapia (Oreochromis niloticus) Broodstock-fed Dietary Fermented Extracts Sourced from Saccharomyces cerevisiae (Hilyses): A Field Trial.

The present study aimed to evaluate the effect of dietary fermented extracts sourced from Saccharomyces cerevisiae (nucleotides, ß-glucans and MOS) (Hilyses®) on the production and health of Nile tilapia (Oreochromis niloticus) broodstock, as well as on seed survival and performance. The trial was performed in a hatchery along the spawning season and continued in the laboratory to monitor the performance in fry and fingerlings. The broodstock were divided into two groups, (C) fed a basal diet and (H) fed 0.4% Hilyses. Blood and histological parameters, antioxidant power, cortisol level and the expression of some immune-related (TLR-2, IL-1ß and TNF-α) and growth-related genes (MUC-2 and IGF-1) were measured. The obtained seeds were subdivided into four treatments: (C-C) fed a basal diet, (C-H) fed 0.4% Hilyses, (H-C) fed a basal diet and (H-H) fed 0.4% Hilyses. Results revealed that the dietary inclusion of Hilyses in the broodstock increased seed production, survival, hematological parameters, and antioxidant power. Moreover, it improved the intestinal microstructure and upregulated the immune- and growth-related genes. The growth indices of fry and fingerlings were significantly increased in all Hilyses-treated groups (p < 0.05). The performance in the (H-H) group significantly surpassed those of all groups. Therefore, dietary fermented yeast could be used as a strategic solution to sustain tilapia production.

Antiparasitic and Antibacterial Functionality of Essential Oils: An Alternative Approach for Sustainable Aquaculture.

Using synthetic antibiotics/chemicals for infectious bacterial pathogens and parasitic disease control causes beneficial microbial killing, produces multi-drug resistant pathogens, and residual antibiotic impacts in humans are the major threats to aquaculture sustainability. Applications of herbal products to combat microbial and parasitic diseases are considered as alternative approaches for sustainable aquaculture. Essential oils (EOs) are the secondary metabolites of medicinal plants that possess bioactive compounds like terpens, terpenoids, phenylpropenes, and isothiocyanates with synergistic relationship among these compounds. The hydrophobic compounds of EOs can penetrate the bacterial and parasitic cells and cause cell deformities and organelles dysfunctions. Dietary supplementation of EOs also modulate growth, immunity, and infectious disease resistance in aquatic organisms. Published research reports also demonstrated EOs effectiveness against Ichthyophthirius multifiliis, Gyrodactylus sp., Euclinostomum heterostomum, and other parasites both in vivo and in vitro. Moreover, different infectious fish pathogenic bacteria like Aeromonas salmonicida, Vibrio harveyi, and Streptococcus agalactiae destruction was confirmed by plant originated EOs. However, no research was conducted to confirm the mechanism of action or pathway identification of EOs to combat aquatic parasites and disease-causing microbes. This review aims to explore the effectiveness of EOs against fish parasites and pathogenic bacteria as an environment-friendly phytotherapeutic in the aquaculture industry. Moreover, research gaps and future approaches to use EOs for sustainable aquaculture practice are also postulated.

Influence of dietary fermented Saccharomyces cerevisiae on growth performance, oxidative stress parameters, and immune response of cultured Oreochromis niloticus.

Our trial was performed to investigate the effect of fully fermented yeast Sacharomyces cerevisiae (Hilyses, ICC Company, Brazil) on the growth performance and immune response of Oreochromis niloticus. In this study, a total of 270 O. niloticus (50.7 ± 0.8 g) were randomly divided into 3 groups in triplicates. The control group was fed on the basal diet while the other two groups were fed on a basal diet supplemented with 0.2% and 0.4% of Hilyses. The trial period extended for 2 months. At the end of the feeding trial, oxidant and antioxidant parameters (MDA, catalase, and glutathione reductase), some innate immunological parameters and immune-related gene expression were measured. Histological examination of liver, spleen, kidney, and intestine was performed. Further, fish groups were challenged against Gram-negative and Gram-positive bacteria; A. hydrophila and L. garvieae. The results revealed significant improvement (p < 0.05) in growth performance and feed utilization in Hilyses-treated groups versus the control group. Blood parameters and liver and kidney functions of Hilyses-supplemented groups were similar to those of the control group. The histological findings of treated groups showed normal tissue structure with multiple focal lymphoid aggregations in the spleen, kidney, and intestine. Both levels of Hilyses successfully enhanced phagocytic activity/index, lysozyme activity, and gene expression of TNF-α, and IL-1ß. Fish group fed on 0.4% Hilyses exhibited the highest expression of IL-1ß and the least mortality percentages post challenges. Thus, dietary supplementation of Hilyses could promote the growth performance and immunity and increase the resistance of O. niloticus against diseases.

Efficacy of Injectable and Immersion Polyvalent Vaccine against Streptococcal Infections in Broodstock and Offspring of Nile tilapia (Oreochromis niloticus).

A vaccine against streptococcosis, lactococcosis and enterococcosis in tilapia was formulated, ME-VAC Aqua Strept, as a polyvalent inactivated vaccine containing Streptococcus agalactiae, S. iniae, Lactococcus garvieae and Enterococcus faecalis along with a nano-particulate adjuvant. Use of ME-VAC Aqua Strept by injection or immersion resulted in an improved non-specific and adaptive immunity of broodstock and offspring. Intra-peritoneal vaccination of tilapia broodstock increased the total leukocyte count, phagocytosis, lysozyme activity, antibody titer, number of seeds/vaccinated broodstock, seeds quality and survival rates. Also, immersion mass vaccination of tilapia larvae provided a long period of protection up to three months, with a relative percent of survivability (RPS) not less than 60% at this time. To our knowledge, this vaccine may be the first to offer a combined protection against streptococcosis, lactococcosis and enterococcosis in tilapia. The results support the use of this vaccine as an effective tool for disease control and well-being of fish.

Aquatic environmental risk assessment of chitosan/silver, copper and carbon nanotube nanocomposites as antimicrobial agents.

Despite the potential antimicrobial and water purification benefits of chitosan-based nanocomposites, there are growing concerns regarding the hazards of leached nanoparticles (NPs) to the in-contact circumference. The antibacterial performance of the nanocomposites of chitosan with silver and copper NPs and carbon nanotubes was assessed with an emphasis on their impact on fish health. The minimal inhibitory concentrations of each preparation and the growth curves of Aeromonas hydrophila exposed to different nanocomposites were measured. Five groups of Oreochromis niloticus were exposed to chitosan nanocomposites for three weeks. A combination of a low concentration of the NPs in the chitosan matrix improved their antimicrobial properties. However, aqueous exposure to these materials still had hazardous effects on fish health. Experimental groups of O. niloticus exposed to these nanocomposites exhibited oxidative stress, tissue DNA fragmentation and higher expression of pro-inflammatory and immune-related genes such as TNF-α and IL1ß. Various pathological tissue alterations were observed in gills, liver, spleen and intestine. Exposure to some of the prepared nanocomposites led to significant DNA damage in hepatic cells with a marked increase in the apoptotic index.

Chitosan-silver nanocomposites in goldfish aquaria: A new perspective in Lernaea cyprinacea control.

Nanomedicine is a promising new research area in human and veterinary science. Metal nanoparticles have shown high biocidal activity against bacteria, fungi and viruses, few studies have focused on antiparasitic action. Therefore, this study aims to investigate the influence of chitosan-silver nanocomposites on the fish crustacean parasite Lernaea cyprinacea. The disease was detected in goldfish (Carassius auratus) aquaria during the spring. Molecular and morphometric characterizations of the parasite were performed using polymerase chain reaction for rRNA and scanning electron microscopy. Chitosan-silver nanocomposites were characterized using transmission electron microscopy and Zetasizer. Probit analysis of parasite mortality versus the logarithmic concentrations of the composites indicated that the 1h/LC50 was 5.495ppm. Parasites exposed to the chitosan-silver nanocomposites showed severe pathological alterations and adsorbed the composite particles on their cuticles. After aqueous exposure of the infected fish to the compound at its LC50 for 24h, the female lernaeids were completely dislodged. Moreover, the pathological findings indicated rapid skin wound healing and renewal at the parasitic injury site. Therefore, we concluded that chitosan-silver nanocomposites are potential parasitic control agents for ornamental glass aquaria, as they have detrimental effects on aquatic predators, such as copepods.

Eutrophication, Ammonia Intoxication, and Infectious Diseases: Interdisciplinary Factors of Mass Mortalities in Cultured Nile Tilapia.

The present study was designed to assess the possible causes of the mass mortalities of Nile Tilapia Oreochromis niloticus at El-Behera Governorate, Egypt, in relationship to environmental and microbiotic factors. Water samples were collected from fish farms at different locations and from Lake Edku to analyze water temperature, water pH, salinity, biological oxygen demand, dissolved oxygen, total ammonia nitrogen, and un-ionized ammonia. A number of moribund and freshly dead fish were sampled and submitted to our laboratory for microbiological, molecular, and histopathological examination. Water analysis of the fish farms revealed noticeable increases in the previously mentioned physicochemical parameters. Clinical examinations of moribund fish showed severe gill rot and massive external and internal hemorrhages. Ordinary and molecular laboratory findings confirmed the presence of Branchiomyces sp. in gill tissue and mixed bacterial fish pathogens (Streptococcus agalactiae, Vibrio alginolyticus, V. parahaemolyticus, Pseudomonas anguilliseptica, and P. aeruginosa) in visceral organs. The histopathological and transmission electron microscopic examinations revealed severe necrosis of gill filaments and blockage of branchial blood vessels and lamellar capillaries with Branchiomyces sp. hyphae and spores mixed with different shapes of bacteria. Severe inflammations were detected in liver, kidney, heart, and brain tissues. Ultimately, we can conclude that the syndrome of mass fish kills in this area is a consequence of ecological damage to the aquatic environment, which is mainly related to natural and anthropogenic factors, as well as to the presence of infectious agents. Received September 30, 2015; accepted April 12, 2016.

Assessment of the immune-modulatory and antimicrobial effects of dietary chitosan on Nile tilapia (Oreochrmis niloticus) with special emphasis to its bio-remediating impacts.

Fish, pathogen and environment are three counterparts who are sharing the same circle of life. To keep fish up to their optimal health, environment should be competently improved and pathogen count/virulence should be seized. Using of bioactive immunostimulants to achieve these objectives is the hypothesis under assessment. Thus, the present study was performed to evaluate the use of shrimp shells derived chitosan as an immunostimulant as well as preventive regime against Aeromonas hydrophila infection of Nile tilapia and to assess its antibacterial/aquatic bio-remediating effects. Results achieved by feeding 1% chitosan as preventive/therapeutic regimes have revealed a remarkably enhanced several innate immunological parameters (e.g., Phagocytic activity/index, NBT, Lysozyme activity and ACH50), increased resistance against A. hydrophila and strikingly improved water quality compared to the 0.5 and 2% chitosan containing diets. Conclusively, experimental results suggest the commercial usage of chitosan as an efficient immunostimulant and bio-remediating agent in aquaculture.

Eubiotic effect of a dietary acidifier (potassium diformate) on the health status of cultured Oreochromis niloticus.

In connection with the global demand for safe human food and the production of environmentally friendly aquaculture products, acidifiers are natural organic acids and salts that have received considerable attention as animal-feed additives. The current study was designed to evaluate the effects of potassium diformate (KDF) on the growth performance and immunity of cultured Oreochromis niloticus (O. niloticus). Four iso-nitrogenous and iso-caloric rations containing graded levels of KDF, including 0% (control basal diet), 0.1%, 0.2% and 0.3%, were fed separately to four equal fish groups (30 fish/group with an initial body weight of 53.49 ± 6.15 g) for sixty days. At the end of the experimental period, the fish groups fed on 0.2% and 0.3% KDF exhibited significant improvements in their feed intake, live weight gain, specific growth rate, feed conversion ratio and protein efficiency ratio, with concomitant improvement of their apparent protein digestibility (p < 0.05). Dietary supplementation of 0.3% KDF appeared to stimulate the beneficial intestinal flora; a proliferation was observed of indigenous probionts (Eubiosis) associated with the relative activation of cellular and humeral innate immunity (phagocytic activity/index, nitroblue tetrazolium reduction test and serum/gut mucous lysozyme activity). The cumulative mortality of the fish groups fed on KDF and challenged orally with Aeromonas hydrophila was lower than that of the control group. The resistance against diseases increased with dietary KDF in a dose-dependent manner. Thus, we conclude that the use of acidifiers can be an efficient tool to achieve sustainable, economical and safe fish production.