Dataset Information

Optimization of the cytogenetic protocol for Pangasianodon hypophthalmus (Sauvage, 1878) and Clarias gariepinus (Burchell, 1822).

ABSTRACT: To obtain well spread chromosomes, the cytogenetic protocol for Pangasianodon hypophthalmus and Clarias gariepinus were optimized. This includes, the colchicine concentration (0.01%, 0.025%, 0.05%)/exposure duration (1, 3, and 5 h), hypotonic solution (distilled water or 0.075M KCl solution)/exposure duration (30 min, 1, and 2 h), the time of cell suspension preparation (at hypotonic treatment or before slide preparation) and chromosome aging period (0, 3, and 7 days in Carnoy's fixative). In addition, the type (i.e., fin, gill or kidney) and the amount of tissue (10, 50, 100 or 150 mg) were also investigated. Regardless of the species, the result obtained showed that well-spread chromosomes could be obtained using the following optimized protocol: Juveniles are injected with 0.05% colchicine (at one ml kg^-1) and allowed to swim for 3 h. Then, 50 mg of gill tissue is made into cell suspension in 0.075M KCl for 1 h. The cell suspension is treated in Carnoy's fixative (changed three times at 20 min interval) and then aged for 3 days. Finally, chromosome slides are made and stained with 10% Giemsa for 1 h.

SUBMITTER: Okomoda VT

PROVIDER: S-EPMC6223236 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Optimization of the cytogenetic protocol for Pangasianodon hypophthalmus (Sauvage, 1878) and Clarias gariepinus (Burchell, 1822).

Okomoda Victor T VT Koh Ivan C C ICC Hassan Anuar A Amornsakun Thumronk T Moh Julia H Z JHZ Shahreza Sheriff Md SM

PeerJ 20181105

To obtain well spread chromosomes, the cytogenetic protocol for Pangasianodon hypophthalmus and Clarias gariepinus were optimized. This includes, the colchicine concentration (0.01%, 0.025%, 0.05%)/exposure duration (1, 3, and 5 h), hypotonic solution (distilled water or 0.075M KCl solution)/exposure duration (30 min, 1, and 2 h), the time of cell suspension preparation (at hypotonic treatment or before slide preparation) and chromosome aging period (0, 3, and 7 days in Carnoy's fi ...[more]

PMID: 30416879

Similar Datasets

Project description:Antimicrobial peptides (AMPs) constitute a broad range of bioactive compounds in diverse organisms, including fish. They are effector molecules for the innate immune response, against pathogens, tissue damage and infections. Still, AMPs from African Catfish, Clarias gariepinus, skin mucus are largely unexplored despite their possible therapeutic role in combating antimicrobial resistance. In this study, African Catfish Antimicrobial peptides (ACAPs) were identified from the skin mucus of African Catfish, C. gariepinus. Native peptides were extracted from fish mucus scrapings in 10% acetic acid (v/v) and ultra-filtered using 5 kDa molecular weight cut-off membrane. The extract was purified using C18 Solid-Phase Extraction. The antibacterial activity was determined using the Agar Well Diffusion method and broth-dilution method utilizing Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922). Thereafter, Sephadex G-25 gel filtration was further utilized in bio-guided isolation of the most active fractions prior to peptide identification using Orbitrap Fusion Lumos Tribrid Mass Spectrometry. The skin mucus extracted from African Catfish from all the three major lakes of Uganda exhibited antimicrobial activity on E. coli and S. aureus. Lake Albert's C. gariepinus demonstrated the best activity with the lowest MIC of 2.84 and 0.71 μg/ml on S. aureus and E. coli, respectively. Sephadex G-25 peak I mass spectrometry analysis (Data are available via ProteomeXchange with identifier PXD029193) alongside in silico analysis revealed seven short peptides (11-16 amino acid residues) of high antimicrobial scores (0.561-0.905 units). In addition, these peptides had a low molecular weight (1005.57-1622.05 Da) and had percentage hydrophobicity above 54%. Up to four of these AMPs demonstrated α-helix structure conformation, rendering them amphipathic. The findings of this study indicate that novel AMPs can be sourced from the skin mucus of C. gariepinus. Such AMPs are potential alternatives to the traditional antibiotics and can be of great application to food and pharmaceutical industries; however, further studies are still needed to establish their drug-likeness and safety profiles.

Project description:Discrimination of different fishes can be done through different means which includes morphological appearance. When two fishes are successfully hybridized, they produce progenies that have shared morphology between their pure parent, hence, making morphometric characterization an important aspect of hybrid discrimination. However, erythrocyte characterization is also a simpler method for characterization. The dataset presented in this article represents the traditional morphological data, truss network data and erythrocyte data of pure and novel hybrids from reciprocal crosses of African catfish Clarias gariepinus and Asian catfish Pangasianodon hypophthalmus. Breeding of the broodstocks was done to produce pure and hybrid progenies which were maintained for a period of four to six months. Based on the cross combinations and morphotypes, traditional measurement of twenty-five morphological characters and five meristic counts were recorded. Thereafter pictures of the different fish groups were used to determine values of thirty-six distances between ten landmark points. The morphological abnormality of the hybrids at market size is also presented in this data article for the very first time. Blood was then collected from the caudal peduncle of ten fish per group and smeared on a slide for observation under a compound microscope (at 100 × magnification). Data gotten included erythrocytes parameters such as cell major axis, cell minor axis, nucleus major axis, nucleus minor axis cell area, nucleus area, cell volume, and nucleus volume. Data recording was through the Microsoft excel spreadsheet; which was also used to process the data to get the exclusive ranges of values for paired progenies. The data as presented is associated with the research article "Morphological characterization of the progenies of pure and reciprocal crosses of Pangasianodon hypophthalmus (Sauvage, 1878) and Clarias gariepinus (Burchell, 1822)" [1]. The dataset presented in this article can be used for easy identification of the novel hybrid progenies of the African Catfish and Asian Catfish.

Project description:The aim of the study was to comparatively analyze the interrelationships among iron (Fe), manganese (Mn), zinc (Zn), copper (Cu), lead (Pb), cadmium (Cd), chromium (Cr) and total hydrocarbons (THCs) in the gills, intestines and muscles of Clarias gariepinus collected from Osse River, Nigeria, between the periods of April, 2013 to September, 2014. The trace metals in the fish tissues were analyzed using Atomic Absorption Spectrophotometer (AAS, Philips model PU 9100), while total hydrocarbons were analyzed using High Performance Liquid Chromatograph (HPLC,Prominence Dual brand from HGE) equipped with a detector Shimadzu UV-Visible (UV-Vis Prominence SPD 20?A). The concentrations of trace metals and THCs in the tissues were subjected to principal component analysis (PCA), in conjunction with hierarchical cluster analysis (HCA), backed up by correlation analysis (CA). In the most prioritized component among the hierarchies of contaminants, characterized as principal component 1, results of communality extractions and rotated component matrices revealed the order of contaminants was Mn > Cu > Zn > Fe > Cr in the intestines, Cr > Cu > THCs > Mn > Fe in the muscle, while Pb > Cr > Fe > Mn was the order in the gills of the fish. Iron inhibited accumulation of the other trace metals in the gills, where its threshold of essentiality was maximal. Noteworthy is the fact that Mn and Cu were the most active components in the muscle and concurrently of excess concentrations in the tissue, which is the major edible part of fish, and constitutes its main body weight, hence holds its nutritional and economic values. High level of variability which occurred in the toxicant profile across the tissues of C. gariepinus is a function of uptake route, varied organ functions and specificity of tissue permeability of the compared organs. The study demonstrated variability in organ accumulation capacity and toxicant's competitiveness irrespective of bioavailability. The study provides data useful for future ecotoxicological studies and safety of consumers of the fish.

Project description:Antimicrobial peptides (AMPs) constitute a broad range of bioactive compounds in diverse organisms, including fish. They are effector molecules for the innate immune response, against pathogens, tissue damage and infections. Still, AMPs from African Catfish, Clarias gariepinus skin mucus are largely unexplored despite their possible therapeutic role in combating antimicrobial resistance. In this study, African Catfish Antimicrobial peptides (ACAPs) were identified from the skin mucus of African Catfish, C. gariepinus. Native peptides were extracted from fish mucus scrapings in 10% acetic acid (v/v) and ultra-filtered using 5kDa molecular cut-off membrane. The extract was purified using C18 Solid Phase Extraction. The antibacterial activity was determined using the Agar Well Diffusion method and broth-dilution method utilizing Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922). Thereafter, Sephadex G-25 gel filtration was further utilized in bio-guided isolation of the most active fractions prior to peptide identification using Orbitrap Fusion Lumos Tribrid Mass Spectrometry. The skin mucus extracted from African Catfish from all the three major lakes of Uganda exhibited antimicrobial activity on E. coli and S. aureus. Lake Albert’s C. gariepinus demonstrated the best activity with the lowest MIC of 2.84 µg/mL and 0.71 µg/mL on S. aureus and E. coli respectively. Sephadex G-25 peak I mass spectrometry analysis alongside in silico analysis revealed seven short peptides (11-15 amino acid residues) of high antimicrobial scores (0.561-0.905 units). In addition, these peptides had a low molecular weight (1005.57-1622.05 Da), and had percentage hydrophobicity above 54%. Up to four of these antimicrobial peptides demonstrated α-helix structure conformation, rendering them amphipathic. The ﬁndings of this study indicate that novel antimicrobial peptides can be sourced from the skin mucus of C. gariepinus. Such antimicrobial peptides are potential alternatives to the traditional antibiotics and can be of great application to food and pharmaceutical industries; however, further studies are still needed to establish their drug-likeness and safety profiles.

Dataset Information

Optimization of the cytogenetic protocol for Pangasianodon hypophthalmus (Sauvage, 1878) and Clarias gariepinus (Burchell, 1822).

Publications

Optimization of the cytogenetic protocol for <i>Pangasianodon hypophthalmus</i> (Sauvage, 1878) and <i>Clarias gariepinus</i> (Burchell, 1822).

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets