Research on the Artemia nauplii microorganism compositions based on metagenomics next-generation sequencing.

Vibrio species represent the predominant and significant pathogen in global marine fish and shellfish aquaculture. Vibrio species are ubiquitously presented in Artemia cyst hatcheries, and their notable colonization in live prey, particularly Artemia nauplii, leads to the transmission of these pathogens into the digestive system of larval organisms, causing serious problems in Vibriosis in marine aquaculture. To eliminate the Vibriospecies in Artemia nauplii, trichloroisocyanuric acid (TCCA) was used for sterilization of the nauplii. In this study, 3 different concentrations, including 0.5 ppm (FA group), 1.0 ppm (FB group) and 1.5 ppm (FC group) of TCCA were used to treat nauplii for 25 min, and then genomic DNA of the different groups were extracted followed by metagenomic next-generation sequencing (NGS). Bioinformatics analysis was applied and the results indicated that Proteobacter constituted the predominant component within each group at the phylum level, albeit accounting for only 58.68% in the FB group, which was significantly lower than in other groups (>86%). The relative abundance of Vibrio species at genus level showed that when compared with the control group, the FB group (15.8%) was reduced by 25.5%. Beta diversity showed differences between the FB group and the other groups, suggesting that treatment with 1.0ppm TCCA for 25 min would obviously reduce the Vibrio in Artemia nauplii. In conclusion, the Vibrio species were significantly reduced after treatment with TCCA, indicating that TCCA might be an alternative to antibiotics used for live food sterilization in marine aquaculture.

Bio-efficacy of insecticidal molecule emodin against dengue, filariasis, and malaria vectors.

Emodin, a compound isolated from Aspergillus terreus, was studied using chromatographic and spectroscopic methods and compound purity (96%) was assessed by TLC. Furthermore, high larvicidal activity against Aedes aegypti-AeA (LC50 6.156 and LC90 12.450 mg/L), Culex quinquefasciatus-CuQ (8.216 and 14.816 mg/L), and Anopheles stephensi-AnS larvae (6.895 and 15.24 mg/L) was recorded. The first isolated fraction (emodin) showed higher pupicidal activity against AeA (15.449 and 20.752 mg/L). Most emodin-treated larvae (ETL) showed variations in acetylcholine esterase, α and ß-carboxylesterases, and phosphatase activities in the 4th instar, indicating the intrinsic differences in their biochemical changes. ETL had numerous altered tissues, including muscle, gastric caeca, hindgut, midgut, nerve ganglia, and midgut epithelium. Acute toxicity of emodin on brine shrimp Artemia nauplii (54.0 and 84.5 mg/L) and the zebrafish Danio rerio (less toxicity observed) was recorded. In docking studies, Emodin interacted well with odorant-binding-proteins of AeA, AnS, and CuQ with docking scores of - 8.89, - 6.53, and - 8.09 kcal mol-1, respectively. Therefore, A. terreus is likely to be effective against mosquito larvicides.

Insecticidal Efficacy of Metarhizium anisopliae Derived Chemical Constituents against Disease-Vector Mosquitoes.

Insecticides can cause significant harm to both terrestrial and aquatic environments. The new insecticides derived from microbial sources are a good option with no environmental consequences. Metarhizium anisopliae (mycelia) ethyl acetate extracts were tested on larvae, pupae, and adult of Anopheles stephensi (Liston, 1901), Aedes aegypti (Meigen, 1818), and Culex quinquefasciatus (Say, 1823), as well as non-target species Eudrilus eugeniae (Kinberg, 1867) and Artemia nauplii (Linnaeus, 1758) at 24 h post treatment under laboratory condition. In bioassays, Metarhizium anisopliae extracts had remarkable toxicity on all mosquito species with LC50 values, 29.631 in Ae. aegypti, 32.578 in An. stephensi and 48.003 in Cx. quinquefasciatus disease-causing mosquitoes, in A. nauplii shows (5.33-18.33 %) mortality were produced by the M. anisopliae derived crude extract. The LC50 and LC90 values were, 620.481; 6893.990 µg/mL. No behavioral changes were observed. A low lethal effect was observed in E. eugeniae treated with the fungi metabolites shows a 14.0 % mortality. The earthworm E. eugeniae mid-gut histology revealed that M. anisopliae extracts had no more harmful effects on the epidermis, circular muscle, setae, mitochondrion, and intestinal lumen tissues than chemical pesticides. By Liquid chromatography mass spectrometry (LC-MS) analysis, camphor (25.4 %), caprolactam (20.68 %), and monobutyl phthalate (19.0 %) were identified as significant components of M. anisopliae metabolites. Fourier transform infrared (FT-IR) spectral investigations revealed the presence of carboxylic acid, amides, and phenol groups, all of which could be involved in mosquito toxicity. The M. anisopliae derived chemical constituents are effective on targeted pests, pollution-free, target-specific, and are an alternative chemical insecticide.

Insecticidal Efficacy of Microbial-Mediated Synthesized Copper Nano-Pesticide against Insect Pests and Non-Target Organisms.

Currently, medical and stored grain pests are major concerns of public health and economies worldwide. The synthetic pesticides cause several side effects to human and non-target organisms. Copper nanoparticles (CuNPs) were synthesized from an aqueous extract of Metarhizium robertsii and screened for insecticidal activity against Anopheles stephensi, Aedes aegypti, Culex quinquefasciatus, Tenebrio molitor and other non-target organisms such as Artemia salina, Artemia nauplii, Eudrilus eugeniae and Eudrilus andrei. The synthesized copper nano-particles were characterized using, UV-vis spectrophotometer, Fourier Transform Infrared Spectroscopy (FTIR), X-Ray Diffraction (XRD), Energy Dispersive X-Ray analysis (EDaX), High Resolution Scanning Electron Microscope (HR-SEM) and Atomic Force Microscope (AFM) analysis. Insects were exposed to 25 µg/mL concentration produced significant mortality against larvae of A. stephensi, A. aegypti, C. quinquefasciatus and T. molitor. The lower toxicity was observed on non-target organisms. Results showed that, M. robertsii mediated synthesized CuNPs is highly toxic to targeted pests while they had lower toxicity were observed on non-target organisms.

Biogenic synthesis of novel platinum-palladium bimetallic nanoparticles from aqueous Annona muricata leaf extract for catalytic activity.

This work reports the fast and effective bio-fabrication of novel platinum-palladium bimetallic nanoparticles (Pt-Pd BNPs) along with their counterparts Pt and Pd monometallic NPs (MNPs) through aqueous Annona muricata leaf extract. The bio-fabrication of the NPs was achieved within 2 h at 100 °C and pH 7 which was established by the occurrence of dark brown color for Pt MNPs and black color for Pd MNPs and Pt-Pd BNPs. NPs were evaluated for their catalytic activity in the reduction of methyl orange (MO), rhodamine-B (rh-B), and methylene blue (MB) textile dyes in presence of sodium borohydride as a reducing agent. Pt-Pd (1:3) BNPs showed higher MO dye degradation (96.84 ± 2.05% in 50 min) followed by Pd MNPs (97.07 ± 1.46% in 60 min), Pt-Pd (3:1) BNPs (97.34 ± 1.17% in 70 min) and Pt-Pd (1:1) BNPs (98.12 ± 1.04% in 80 min). Pd MNPs showed significant catalytic activity in the reduction of rh-B dye by 97.27 ± 1.14% in 12 min followed by Pt-Pd (3:1) BNPs (96.76 ± 2.17% in 18 min), Pt-Pd (1:3) BNPs (96.53 ± 1.97% in 33 min) and Pt-Pd (1:1) BNPs (97.11 ± 2.09% in 39 min). Pt-Pd (1:3) BNPs also showed higher MB dye degradation (96.95 ± 1.57% in 40 min) followed by Pd MNPs (96.22 ± 2.36% in 55 min), Pt-Pd (3:1) BNPs (97.29 ± 1.22% in 75 min) and Pt-Pd (1:1) BNPs (96.45 ± 2.19% in 105 min). However, Pt MNPs showed no catalytic activity. Standard disc diffusion method was used to evaluate the NPs toxicity towards Escherichia coli and Staphylococcus aureus, which showed no inhibitory zones. NPs showed less toxicity compared to potassium dichromate (control) against Artemia nauplii. Among the NPs studied, Pt-Pd (1:1) BNPs showed less toxicity with 100% mortality only at 100 µg/mL concentration followed by Pt MNPs (≥ 80 µg/mL), Pt-Pd (1:3) BNPs (≥ 60 µg/mL), Pt-Pd (3:1) BNPs (≥ 60 µg/mL) and Pd MNPs (≥ 40 µg/mL) after 72 h exposure. These evaluations support the application of bio-fabricated Pt-Pd BNPs as nano-catalysts in textile dyes degradation. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02935-0.

Chemical and morphological defenses of Pseudo-nitzschia multiseries in response to zooplankton grazing.

Pseudo-nitzschia species frequently blooms in coastal waters, and some species are able to produce the toxin domoic acid (DA), hereby causing harm to the marine ecosystem and humans. Laboratory studies were conducted to investigate the influence of different levels of grazing pressure on the morphological and chemical response (in terms of cellular DA production) of Pseudo-nitzschia. Subsequently, zooplankton grazer responses to these defenses were examined. The cellular DA content of P. multiseries ranged from 0.11-0.27 pg cell-1 without grazers, and increased up to 44% with the presence of grazers (Artemia nauplii) and with grazer concentration. Grazing also affected the density of P. multiseries chains and average chain length which became ~25% higher and ~8% longer, respectively, than without grazers. These effects could either be caused by size-dependent grazing or by grazer-cue-induced effects on chain formation. A negative correlation between cellular DA content in P. multiseries and clearance and/or ingestion rates of Artemia nauplii indicate that DA might have a negative effect on the grazing of Artemia nauplii. Such interaction might result in a decrease in grazing pressure on toxic blooming species, like P. multiseries, and hence potentially a prolonged bloom. This indicates that the interaction between toxic diatoms and grazers may have implications on aquatic food web structure and the progression of Pseudo-nitzschia blooms.

Synergistic effect of durian fruit rind polysaccharide gel encapsulated prebiotic and probiotic dietary supplements on growth performance, immune-related gene expression, and disease resistance in Zebrafish (Danio rerio).

The present study investigated the effect of polysaccharide gel (PG) extracted from the rind of durian fruit which is encapsulated with Bacillus subtilis as a feed and co-inoculation with Artemia nauplii in the induction of immune response in Danio rerio after Vibrio immersion challenge (5 days). The total RBC count, lysozyme activity test, weight, and length analysis revealed that the zebra fishes fed with the PG encapsulated probiotics had more immune induction activity, survival, and growth than the non-encapsulated groups. When the expression of the immune-related genes was measured, the studies revealed the significant upregulation (P < 0 .05) of interleukin 1 beta (Il1b), lysozyme (lyz), tumor necrosis factor-alpha (TNF-α), superoxide dismutase (SOD) genes in fish fed with PG encapsulated probiotics with A. nauplii showed an immense effect on the induction of immune response compared to other feeds.

Toxicological evaluation of biosynthesised hematite nanoparticles in vivo.

In recent years, nanomaterials have been widely used in consumer products. High reactivity of metallic nanoparticles and its bioaccumulation in biological systems are the main causes of concern over their safety to human health and environment. The available information related to the safety of several nanomaterials is insufficient. Hematite nanoparticles are proposed for various applications. Ecotoxicological studies of hematite nanoparticles are very limited. In the present study, biosynthesised hematite nanoparticles using Bacillus cereus were evaluated for its acute oral toxicity in mice following OECD guidelines. A dose of 2 g/kg/p.o was administered to Swiss albino mice through gastric oral feeding tube and observed for 14 days. After two weeks blood samples were collected and subjected for evaluation of haematological parameters and biochemical analysis. There was no mortality and toxic signs of animals till the end of observational period. The animals were sacrificed and organs like liver and kidneys were isolated to study the histopathological changes. The results of the study revealed that there was no drastic change in parameters except slight change in bilirubin in the hematite nanoparticle treated mice. Biosynthesised hematite nanoparticles were assayed for toxicity in Artemia salina. Cysts treated with higher concentrations of hematite nanoparticles showed small sized nauplii. Biosynthesised hematite nanoparticles were found to be non-toxic to A. salina nauplii in lower concentrations.

In vitro and in vivo efficacy of methyl oleate and palmitic acid against ESBL producing MDR Escherichia coli and Klebsiella pneumoniae.

PURPOSE: Antibiotic resistance is a huge problem that stays to challenge the healthcare sector in a large part of the world in both developing and developed countries. The spread of multi drug resistant (MDR) bacteria in hospital and community settings remains a widely uncertain problem and a heavy burden to health services. METHODS: This study unveils the in vitro and in vivo anti-ESBL potential of Methyl oleate (MO) and Palmitic acid (PA) against ESBL producing MDR bacterial pathogens such as Escherichia coli and Klebsiella pneumoniae. Microscopic observations unveiled the anti-ESBL efficacy of test compounds. MTT assay, in vivo anti-infective efficiency of MO and PA was tested with different concentrations. RESULTS: The pure compounds of MO and PA from Oxynema thaianum demonstrated high inhibitory activity in MIC and MBC assays against MDR E. coli and K. pneumoniae. Moreover, the anti-ESBL potential of MO and PA was validated through light, confocal laser scanning and scanning electron microscopic analyses. The IC50 values of MO and PA against A549 cells was recorded as 625 µg mL-1 and 514 µg mL-1, respectively. In Artemia nauplii cytotoxicity assay, the LC50 value of MO and PA were recorded as 53.33 µg mL-1 and 50 µg mL-1 respectively. The 96 h lethal concentrations obtained for Lobeo rohita treated with different concentrations of Methyl oleate and Palmitic acid. The LC50 for MO and PA was 50 mg L-1 and 100 mg L-1, respectively. CONCLUSION: Therefore the study concluded that the promising effects of MO and PA can be used as an alternative biological agent which could be positively explored to treat ESBL producing MDR pathogens.

Estratégias alimentares na larvicultura do peixe ornamental amazônico acará-severo (Heros severus) (Heckel, 1840) / Feeding strategies on larval rearing of severum (Heros severus) (Heckel, 1840), Amazonian ornamental fish

Com o presente estudo, objetivou-se avaliar o tempo de fornecimento de náuplios de artêmia e o período de transição alimentar para pós-larvas de acará-severo (Heros severus). Foram utilizadas 450 pós-larvas de acará-severo, distribuídas em 45 aquários, em delineamento inteiramente ao acaso, em esquema fatorial 3x5, com três repetições. Foram avaliados três períodos de fornecimento de náuplios de artêmia: zero, cinco e 10 dias. Além disso, foram testados cinco períodos de transição alimentar: um, dois, três, quatro e cinco dias, quando os animais receberam alimentação conjunta de náuplios de artêmia e dieta farelada. Ao final do experimento, os peixes foram contados, pesados e fotografados, para avaliação do desempenho produtivo e das variáveis morfométricas. Para todos os parâmetros avaliados, não foi constatada interação entre o tempo de fornecimento de artêmia e o período de transição alimentar. Os animais alimentados por 10 dias com náuplios de artêmia apresentaram os melhores resultados de desempenho e as maiores variáveis morfométricas. Os peixes que receberam alimentação conjunta por três dias apresentaram desenvolvimento satisfatório. Dessa forma, recomendam-se 10 dias de fornecimento de náuplios de artêmia e três dias de transição alimentar, antes de se iniciar o fornecimento exclusivo de dieta inerte para pós-larvas de acará-severo.(AU)

The aim of this study was to evaluate the supply time of artemia and the period of food transition to post-larvae of severum (Heros severus). A total of 450 post-larvae of severum were distributed in 45 aquaria, in a completely randomized design, in a 3x5 factorial scheme, with three replicates. Three feeding periods with artemia nauplii were evaluated: zero, five, and ten days. In addition, five feeding transitions were tested: one, two, three, four and five days, when the animals received joint feeding of artemia nauplii and dry diet. At the end of the experiment the fish were counted, weighed and photographed, to evaluate the productive performance and the morphometric variables. For all parameters evaluated, no interaction between the time of artemia supply and the feeding transition period was observed. Animals fed for 10 days with artemia nauplii showed the best performance results and the highest morphometric variables. The fish that received joint feeding for three days presented satisfactory development. Thus, 10 days of artemia nauplii supply and three days of feeding transition is recommended before starting the exclusive supply of diet inert to post-larvae of severum.(AU)

In vitro and in vivo toxicity assessment of selenium nanoparticles with significant larvicidal and bacteriostatic properties.

In the present study, we investigated the larvicidal and bacteriostatic activity of biosynthesized selenium nanoparticles using aqueous berry extract of Murraya koenigii (Mk-Se NPs). The synthesized Mk-Se NPs were characterized using UV-visible spectroscopy, X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, transmission electron microscopy (TEM), scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) analysis. XRD analysis revealed the crystalline nature of Mk-Se NPs as hexagonal. The FTIR spectra of Mk-Se NPs exhibited a strong peak at 3441 cm-1 corresponding to the OH group. SEM and TEM analysis showed that the Mk-Se NPs were spherical in shape with a size between 50 and 150 nm. EDX peaks confirm the presence of 73.38% of selenium and 26.62% of oxide in Mk-Se NPs. Mk-Se NPs showed significant larvicidal property against the 4th instar larvae of a dengue fever-causing vector Aedes aegypti with LC50- - 3.54 µg mL-1 and LC90- - 8.128 µg mL-1 values. Mk-Se NPs displayed anti-bacterial activity against Gram-positive (Enterococcus faecalis &Streptococcus mutans) and Gram-negative (Shigella sonnei &Pseudomonas aeruginosa) bacteria at 40 and 50 µg mL-1. In addition, Mk-Se NPs reduced bacterial biofilm thickness extensively at 25 µg mL-1. The high antioxidant property at 50 µg mL-1 and low hemolysis activity till 100 µg mL-1 proved the biocompatible nature of Mk-Se NPs. In vitro and in vivo toxicity assessment of Mk-Se NPs showed low cytotoxicity against RAW 264.7 macrophages and Artemia nauplii. Together, our results suggest the potential application of Mk-Se NPs as a nano-biomedicine.

Aspergillus terreus (Trichocomaceae): A Natural, Eco-Friendly Mycoinsecticide for Control of Malaria, Filariasis, Dengue Vectors and Its Toxicity Assessment Against an Aquatic Model Organism Artemia nauplii.

Vector-borne diseases like malaria, filariasis, and dengue are transmitted by mosquitoes and they cause global mortality and morbidity due to an increased resistance against commercial insecticides. The present study was aimed to evaluate the neurobehavioral toxicity, knock-down effect, histopathology, ovicidal, adulticidal, and smoke toxicity effect of Aspergillus terreus extract against three mosquito species, namely Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti (Diptera: Culicidae). The isolated fungal strain was identified as A. terreus (GenBank accession no: KX694148.1) through morphological and molecular (phylogenetic) analysis. The morphological changes in the treated fourth instar larvae shown the demelanization of cuticle and shrinkage of the internal cuticle of anal papillae. The time duration of extract exposure against the larvae determines the level of toxicity. The extract treated larvae were displayed excitation, violent vertical and horizontal movements with aggressive anal biting behavior as the toxic effect on the neuromuscular system. The results of the biochemical analysis indicated that a decrease in the level of acetylcholinesterase, α-carboxylesterase, and ß-carboxylesterase in extract treated fourth instar larvae of all tested mosquito species. The findings of histopathological investigation shown the disorganization of the abdominal region, mainly in mid, hindgut, and gastric caeca, loss of antenna, lateral hair, caudal hair, upper and lower head hairs in the mycelium extract treated An. stephensi, Cx. quinquefasciatus, and Ae. aegypti. The ovicidal bioassay test results showed the mosquito hatchability percentage was directly related to the concentrations of mycelium extract. Nil hatchability of mosquito eggs was noticed at 500 µg/ml concentration. The adulticidal activity of fungal mycelia ethyl acetate extract resulted in a dose-dependent activity (15 and 30 min recovery periods). The higher concentration of extract (1000 mg/L) acted as a repellent, the adult mosquitoes showed restless movement, uncontrolled/anesthetic flight at last died. The better adulticidal activity was observed in the ethyl acetate extract against An. stephensi, Cx. quinquefasciatus followed by Ae. aegypti with the best score of LD50 and LD90 values and nil mortality was found in the control. The results of smoke toxicity assay of the mycelia extract exhibited significant mortality rate against Ae. aegypti (91%), Cx. quinquefasciatus (89%), and An. stephensi (84%). In addition, the present investigation reported the stability and toxic effects of A. terreus mycelium ethyl acetate extract on Artemia nauplii. The swimming speed (0.88 mm s-1) of A. terreus was reduced with ethyl extract 24 h treatment whereas, the control A. nauplii showed the normal speed of 2.96 mm s-1. Altered behavior and swimming movement were observed in the 8 h A. terreus mycelium extract treated A. nauplii. A pale yellow color substance (metabolites) was found in the mid-gut region of the mycelial extract exposed A. nauplii. The outcome of the present study, suggest that the A. terreus metabolites might serve as an alternative, cost-effective, eco-friendly, and target specific mosquitocidal agent in the future.

Interactions among yeast and probiotic bacteria enhance probiotic properties and metabolism offering augmented protection to Artemia franciscana against Vibrio anguillarum.

The interactions of the probiotics Bacillus subtilis, Lactococcus lactis and Lactobacillus plantarum with the yeast Saccharomyces cerevisiae were examined in terms of probiotic and biochemical characteristics. Yeast supernatant had a positive effect on the aggregation biofilm formation capacity and hydrophobicity of probiotics, and resulted in increased lactic acid levels, reduced pH values as well as lower RS and FAN levels of probiotics. The effect of probiotics supernatants on yeast was more complex but best results were obtained in the yeast: probiotic CFS ratio of 1:2 for B. subtilis and of 2:1 for the other probiotics. The observed effects depended on the volume ratio of the cell free supernatant to the culture it was applied on. Best results were obtained by the volume ratio probiotic: yeast of (2:1) for B. subtilis and of (1:2) probiotic: yeast for L. plantarum and L. lactis. These ratios were used for further evaluation in vitro against V. anguillarum, resulting in reduced survival and attachment properties of the pathogen. Moreover, the administration of the corresponding combination of bacteria and yeast to Artemia nauplii greatly improved their survival following a challenge with the pathogen. Our results demonstrate that yeast enhances the protective effect of probiotics in a strain specific manner.

Larvicidal, Histopathological Efficacy of Penicillium daleae against Larvae of Culex quinquefasciatus and Aedes aegypti Plus Biotoxicity on Artemia nauplii a Non-target Aquatic Organism.

Mosquitoes can transmit the terrible diseases to human beings. Soil-borne fungal products act as potential source for low-cost chemicals, used for developing eco-friendly control agents against mosquito-vector borne diseases. The prime aim of study was to check the larvicidal potential of fungus mycelia (by ethyl acetate solvent) extract from Penicillium daleae (KX387370) against Culex quinquefasciatus and Aedes aegypti and to test the toxicity of brine shrimp Artemia nauplii, by observing the physiological activity. The ethyl acetate extract of P. daleae mycelia (after 15 days) from Potato dextrose broth (PDB) medium revealed better result with least LC50 and LC90 values of I-IV instars larvae of Cx. quinquefasciatus (LC50 = 127.441, 129.087, 108.683, and 93.521; LC90 = 152.758, 158.169, 139.091, and 125.918 µg/ml) and Ae. aegypti (LC50 = 105.077, 83.943, 97.158, and 76.513; LC90 = 128.035, 106.869, 125.640, and 104.606 µg/ml) respectively. At higher concentration (1000 µg/ml) of extracts, mortality begins at 18 h of exposure and attained 100% mortality after 48 h exposure. Overall, the activity was depends on the dose and time of exposure to the extracts. The stereomicroscopic and histopathological analysis of Ae. aegypti and Cx. quinquefasciatus larvae treated with mycelium ethyl acetate extract showed complete disintegration of abdominal region, particularly the midgut and caeca, loss of cuticular parts and caudal hairs. Morphological characterization of the fungi was performed and taxonomically identified through 5.8s rDNA technique. The phylogenetic analysis of rDNA sequence was carried out to find out the taxonomic and the evolutionary sketch of isolate in relation to earlier described genus Penicillium. Behavior and swimming speed alteration was analyzed together with mortality. The results of the experiment indicates that swimming behavior recorder (SBR) is a appropriate tool to detect individual swimming speed of the A. nauplii organisms, since the values have been obtained in accordance with control monitored results showed the 2.75 mm s-1 and after 24 h treated found to be 0.72 mm s-1, respectively. The extract-exposed to A. nauplii showed changes in body structures, i.e., intestine enlargement, eye formation, outer shell malformations and loss of antennae. In the present study, we aimed to investigate the toxicity of the ethyl acetate extract of P. daleae on A. nauplii larvae by performing the mortality, behavior and alterations in swimming responses. This is the first time report on the larvicidal efficacy of P. daleae ethyl acetate extract against Cx. quinquefasciatus and Ae. aegypti larvae.

Molecular cloning and expression analysis of the retinoid X receptor (RXR) gene in golden pompano Trachinotus ovatus fed Artemia nauplii with different enrichments.

The retinoid X receptors (RXRs) are involved in the skeletal development and other biological process such as blood vessel formation and metabolism. Partial sequences of RXRα and ß genes were obtained, and their expressions were quantified on golden pompano Trachinotus ovatus at 28 days post hatching (DPH) to explore the molecular response to nutritional manipulation in fish larvae. As live food, Artemia nauplii were separately enriched with Nannochloropsis and Algamac 3080 and non-enriched Artemia nauplii (control) for fish feeding. The expressions of RXRs were detected in the embryos and fish larvae at early stages, suggesting that the skeletal development in golden pompano initiated before yolk re-sorption completion. Fish fed non-enriched Artemia nauplii ended up with higher jaw malformation. The highest specific growth rate was obtained when fish were fed with the Artemia nauplii enriched with Algamac 3080, and the lowest growth rate was observed when fish were fed with unenriched Artemia nauplii. The highest survival was obtained when fish were fed with non-enriched or Nannochloropsis-enriched Artemia nauplii. This study indicates that the use of enriched formula for Artemia nauplii can significantly affect the expression levels of RXRs and jaw malformation of golden pompano larvae, but there is no clear correlation between RXRs expressions and malformation rates when fish are subjected to nutrient challenge.

Sudden weaning of angel fish pterophyllum scalare (Lichtenstein) (Pisces; Cichlidae) larvae from brine shrimp (Artemia sp) nauplii to formulated larval feed.

This study investigated the effects of sudden weaning of angel fish larvae (Pteraphylum scalari) from Artemia nauplii to commercial larval feed. Four days post hatch (DPH) larvae were reared in four different weaning protocols (TR1-TR4) with triplicates in a complete randomize design. Larvae in TR1 and TR4 were exclusively fed Artemia nauplii and dry feed respectively. In TR2 and TR3, larvae were initially fed Artemia nauplii and suddenly wean to formulated feed on 14 DPH and 7 DPH respectively. The experiment was lasted for 28 days. At the end of the experiment, final mean weight (FW), total length (FL), height (FH), Daily Weight Gain (DWG), Specific Growth Rate (SGR), survival and stress index were compared. Significantly highest (P < 0.05) FW, DWG and SGR were observed in TR1 and TR2 while former values of TR3 were not significantly different from TR1. Highest FL observed in TR1 and TR2 while FL of TR2 was statistically similar to that of TR3. The poorest growth was observed in larvae solely fed formulated feed. Survival and the stress index were independent from weaning methods. Although sudden weaning is possible on 7 DPH, larvae showed comparatively higher growth when switch off to formulate feed on 14 DPH.