Percentage Incidences of Bacteria in Mahseer (Tor putitora), Silver carp (Hypophthalmichthys molitrix) Fish Collected from Hatcheries and Local Markets of District Malakand and Peshawar of Khyber Pakhtunkhwa, Pakistan

Fish is the main source of animal protein for human diet. The aim of this study was to find out prevalence of pathogenic bacteria of two selected economically important fish of Pakistan namely Mahseer (Tor putitora) and Silver carp (Hypophthalmichthys molitrix). Live fish samples from hatcheries and dead fish samples from different markets of study area were randomly collected. The fish samples were analyzed for isolation, identification and prevalence of bacteria. The isolated bacteria from study fish were identified through biochemical test and about 10 species of pathogenic bacteria were identified including the pathogenic bacteria to human and fish namely, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus iniae, Serratia spp. Citrobacter spp. Stenotrophomonas spp. Bacillus spp. and Salmonella spp. The bacterial percentage frequency of occurrence in Silver carp and Mahseer fish showed Pseudomonas aeruginosa 21.42%, Staphylococcus epidermidis 17.85%, Escherichia coli 11.90%, Staphylococcus aureus 9.52%, Citrobacter spp. 9.52%, Serratia spp. 8.33%, Streptococcus iniae 7.14%, Stenotrophomonas spp. 5.95%, Bacillus spp. 4.76% and Salmonella spp. 3.57%. The study revealed that Fish samples of Mahseer and Silver carp that were collected from markets have found more isolates (10 bacterial species) than did the fresh fish pond samples (03 bacterial species) of hatcheries. The occurrence of pathogenic bacteria in study fish showed risk factor for public health consumers.

Molecular Characterization and Response of Prolyl Hydroxylase Domain (PHD) Genes to Hypoxia Stress in Hypophthalmichthys molitrix

As an economically and ecologically important freshwater fish, silver carp (Hypophthalmichthys molitrix) is sensitive to low oxygen tension. Prolyl hydroxylase domain (PHD) proteins are critical regulators of adaptive responses to hypoxia for their function of regulating the hypoxia inducible factor-1 alpha subunit (HIF-1α) stability via hydroxylation reaction. In the present study, three PHD genes were cloned from H. molitrix by rapid amplification of cDNA ends (RACE). The total length of HmPHD1, HmPHD2, and HmPHD3 were 2981, 1954, and 1847 base pair (bp), and contained 1449, 1080, and 738 bp open reading frames (ORFs) that encoded 482, 359, and 245 amino acids (aa), respectively. Amino acid sequence analysis showed that HmPHD1, HmPHD2, and HmPHD3 had the conserved prolyl 4-hydroxylase alpha subunit homolog domains at their C-termini. Meanwhile, the evaluation of phylogeny revealed PHD2 and PHD3 of H. molitrix were more closely related as they belonged to sister clades, whereas the clade of PHD1 was relatively distant from these two. The transcripts of PHD genes are ubiquitously distributed in H. molitrix tissues, with the highest expressional level of HmPHD1 and HmPHD3 in liver, and HmPHD2 in muscle. After acute hypoxic treatment for 0.5 h, PHD genes of H. molitrix were induced mainly in liver and brain, and different from HmPHD1 and HmPHD2, the expression of HmPHD3 showed no overt tissue specificity. Furthermore, under continued hypoxic condition, PHD genes exhibited an obviously rapid but gradually attenuated response from 3 h to 24 h, and upon reoxygenation, the transcriptional expression of PHD genes showed a decreasing trend in most of the tissues. These results indicate that the PHD genes of H. molitrix are involved in the early response to hypoxic stress, and they show tissue-specific transcript expression when performing physiological regulation functions. This study is of great relevance for advancing our understanding of how PHD genes are regulated when addressing the hypoxic challenge and provides a reference for the subsequent research of the molecular mechanisms underlying hypoxia adaptation in silver carp.

The Effect of Salt on the Gelling Properties and Protein Phosphorylation of Surimi-Crabmeat Mixed Gels

The effects of different salt additions (1.0%, 1.5%, 2.0%, 2.5%, 3.0%, and 3.5%) on the gelling properties and protein phosphorylation of the mixed gels (MG) formed by silver carp (Hypophthalmichthys molitrix) surimi with 10% crabmeat were investigated. The MG’s breaking force, deformation, gel strength, and water-holding capacity (WHC) increased as the salt concentration increased. The intrinsic fluorescence intensity of the samples initially decreased and then increased, reaching the lowest when the NaCl concentration was 2.5%. The result of SDS–polyacrylamide gel electrophoresis indicated that large aggregates were formed by protein–protein interaction in the MG containing 2.5% or 3.0% NaCl, decreasing the protein band intensity. It was also found that with the addition of NaCl, the phosphorus content initially increased and then decreased, reaching the maximum when the NaCl concentration was 2% or 2.5%, which was similar to the changing trend of actin band intensity reported in the results of Western blot. These results revealed that the amount of salt used had a significant effect on the degree of phosphorylation of the MG protein. The increase in phosphorylation was linked to improved gelling properties, which could lead to new ideas for manufacturing low-salt surimi products in the future.

Diluted Acetic Acid Softened Intermuscular Bones from Silver Carp (Hypophthalmichthys molitrix) by Dissolving Hydroxyapatite and Collagen

Intermuscular bones (IBs) pose physical hazards that threaten consumer health and food safety. This study aimed to investigate the mechanism of softening IBs from silver carp with diluted acetic acid. IBs (separated from muscle) and fillets (without removing IBs) were treated with diluted acetic acid. Analyses of sensory attributes and the hardness of treated IBs indicated that diluted acetic acid (<10 mmol/L) could soften IBs effectively. Additionally, 0.5 mmol/L acetic acid softened IBs within fillets without significantly affecting the texture and flavor of fillets. Analyses of microstructure, minerals (calcium and phosphorus) and collagen content, and the Fourier transform infrared (FTIR) spectra of IBs indicated that acetic acid broke connections (formed by collagen that shared hydroxyl groups) between collagen molecules, and between collagen and hydroxyapatite (HAP), thus inducing the dissolution of collagen and HAP. The dissolution of HAP contributed more to IBs softening than collagen.

Changes in Gel Structure and Chemical Interactions of Hypophthalmichthys molitrix Surimi Gels: Effect of Setting Process and Different Starch Addition

The modifications of histological properties and chemical forces on heated surimi gels with starch addition (0–12 g/100 g surimi) were investigated. Two types of heating processes (direct heating and two-step heating) were carried out on surimi gels in order to reveal the effect of setting on mixed matrices. The results of transverse relaxation time showed less immobile water and free water converted into bound water in a matrix subjected to the setting process. Scanning electron microscope and light microscopy images revealed inefficient starch-swelling in two-step heated gels. Chemical interactions and forces in direct cooking gels were more vulnerable to starch addition, resulting in significant decreases in hydrophobic interaction and sulfhydryl content (p < 0.05). With the increment of starch, the disulfide stretching vibrations of the gauche–gauche–gauche conformation were reduced in both gel matrices. The structural variations of different components collectively resulted in changes in texture profile analysis and water holding capacity. Overall, the results demonstrated that starch addition had a great and positive effect on the weak gel matrix by direct heating.