Impact of antioxidant potential of rohu (Labeo rohita) swim bladder gelatin hydrolysate on oxidative stability, textural and sensory properties of fish sausage enriched with polyunsaturated fatty acids.

The impact of rohu swim bladder gelatin hydrolysate (SBGH) at different levels on textural, sensory, oxidative, and microbial properties of polyunsaturated fatty acids enriched rohu fish cooked sausages (PUFA-RFS) were investigated in the current study. SBGH addition enhanced the lightness values of PUFA-RFS compared to both control sausages (without SBGH and with butylated hydroxyanisole (BHA) (P > 0.05). PUFA-RFS added with 3% SBGH exhibited higher hardness, cohesiveness, and gumminess throughout the storage duration at both 4 °C and -20 °C temperatures when compared to other sausages counterparts. PUFA-RFS added with SBGH displayed lower PV, TBARS, and total microbial counts than the control sausages. Furthermore, PV, TBARS, and total microbial count values of sausage decreased with an increase in SBGH level, indicating retardation in lipid oxidation and microbial growth by SBGH in a dose-depended manner. Nevertheless, sausage added with 3% SBGH had higher overall acceptability than other sausage counterparts. Therefore, SBGH could retard lipid oxidation and improves textural properties of PUFA-enriched fish sausage.

Production of Antioxidant, Angiotensin-Converting Enzyme Inhibitory and Osteogenic Gelatin Hydrolysate from Labeo rohita Swim Bladder.

Optimization of antioxidants and angiotensin-converting enzyme (ACE) inhibitory potential gelatin hydrolysate production from Labeo rohita (rohu) swim bladder (SBGH) by alcalase using central composite design (CCD) of response surface methodology (RSM) was investigated. The maximum degree of hydrolysis (DH), 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS), total antioxidants (TAO), and ACE inhibitory activity were achieved at 0.1:1.0 (w/w) enzyme to substrate ratio, 61 °C hydrolysis temperature, and 94-min hydrolysis time. The resulting SBGH obtained at 19.92% DH exhibited the DPPH (24.28 µM TE/mg protein), ABTS (34.47 µM TE/mg protein), TAO (12.01 µg AAE/mg protein), and ACE inhibitory (4.91 µg/mg protein) activity. Furthermore, SBGH at 100 µg/ml displayed osteogenic property without any toxic effects on MC3T3-E1 cells. Besides, the protein content of rohu swim bladder gelatin (SBG) and SBGH was 93.68% and 94.98%, respectively. Both SBG and SBGH were rich in glycine, proline, glutamic acid, alanine, arginine, and hydroxyproline amino acids. Therefore, SBGH could be an effective nutraceutical in functional food development.

Understanding feeding competition under laboratory conditions: Rohu (Labeo rohita) versus Amazon sailfin catfish (Pterygoplichthys spp.).

Competitive interactions between species is widely prevalent within the animal world. In this manuscript, we attempted to understand feeding competitions between the Amazon sailfin catfish, an invasive species introduced globally, and rohu, a keystone species native to several countries within southeast Asia. We used two different size classes of each species, large-size having total length (TL, from snout tip to caudal fin) of 15-20cm and fingerling having TL<6cm, and feeding duration was used as a proxy to understand competition. Our results demonstrated that feeding durations of large-size rohu were either similar or significantly (P<0.05) higher in presence of catfish when compared to trials in presence of conspecifics, indicating that large-size rohu is not a weak competitor. However, feeding durations of fingerling rohu was significantly (P<0.05) reduced in presence of both large-size and fingerling catfish, when compared to trials in presence of conspecifics. Moreover, fingerling rohu also displayed freeze (alarm) behavior in presence of the catfish. Interestingly, presence of rohu had no significant (P>0.05) impact on feeding durations of catfish. Overall, the study demonstrated that invasive catfish may behaviorally outcompete fingerling rohu, thus, threatening the sustenance of a species that is native to several freshwaters around the globe.

Xylooligosaccharide supplementation in rice protein concentrate based diets: A comprehensive analysis of performance and health of Labeo rohita.

The primary aim of this study was to examine the impact of xylooligosaccharide (XOS) in rice protein concentrate (RPC) based diets on the growth performance, body composition, digestive enzymes, intestinal morphology and blood biochemistry of Labeo rohita fingerlings. Four different XOS levels (0%, 0.5%, 1% and 2%) were used at each RPC (75% and 100%) level. Twenty-five fish per tank with an average initial weight of 25 ± 0.05 g were randomly assigned (Randomised complete block design) to each of the 8 groups in triplicate aquaria (36 × 16 × 12″) and then fed with respective diets @ 3% body weight for 90 days. The results showed significant improvements in growth performance, such as increased weight gain %, specific growth rate, and protein efficiency ratio and improved feed conversion ratio in 1% XOS supplemented diet at 75% RPC. A significant decrease in serum alkaline phosphatase activity (ALP) and plasma melanodialdehyde (MDA) were observed at 1% XOS level in 75% RPC based diets, respectively. Meanwhile, the lowest total cholesterol and highest lysozyme activity were observed in 1% XOS supplemented diet at 75% RPC levels. Moreover, the serum (alanine aminotransferase and aspartate transaminase) and plasma (superoxide dismutase, triglyceride, high density and low density lipoprotein) activities showed nonsignificant effects among the treatments. Furthermore, the digestive enzymes (protease & lipase) and intestinal morphology were significantly influenced at 1% XOS in the 75% RPC-based diet. Polynomial regression analysis showed that 1.25% XOS is the optimum requirement for the growth of rohu fingerlings when fed at 75% RPC based diets. Overall, it was concluded that the 75% RPC diet was efficiently replaced by fishmeal along with 1% XOS addition in L. rohita fingerlings without any negative effect on growth performance and intestinal health.

Diffusion of organochlorine (OCPs) and cypermethrin pesticides from rohu (Labeo rohita) internal organs to edible tissues during ice storage: a threat to human health.

The migration of organochlorine pesticides (OCPs) and cypermethrin residues from internal organs to edible tissues of ice-held Labeo rohita (rohu) was investigated in this study. The liver (246 µg/kg) had the highest level of ∑OCP residues, followed by the gills (226 µg/kg), intestine (167 µg/kg), and muscle tissue (54 µg/kg). The predominant OCPs in the liver and gut were endosulfan (53-66 µg/kg), endrin (45-53 µg/kg), and dichloro-diphenyl-trichloroethane (DDT; 26-35 µg/kg). The ∑OCP residues in muscle increased to 152 µg/kg when the entire rohu was stored in ice, but they decreased to 129 µg/kg in gill tissues. On days 5 and 9, the total OCPs in the liver increased to 317 µg/kg and 933 µg/kg, respectively. Beyond day 5 of storage, total internal organ disintegration had led to an abnormal increase in OCP residues of liver-like mass. Despite a threefold increase in overall OCP residues by day 9, accumulation of benzene hexachloride (BHC) and heptachlor was sixfold, endrin and DDT were fourfold, aldrin was threefold, and endosulfan and cypermethrin were both twofold. Endosulfan, DDT, endrin, and heptachlor were similarly lost in the gills at a rate of 40%, while aldrin and BHC were also lost at 60 and 30%, respectively. The accumulation of OCP residues in tissues has been attributed to particular types of fatty acid derivatives. The study concluded that while pesticide diffusion to edible tissues can occur during ice storage, the levels observed were well below the allowable limit for endosulfan, endrin, and DDT.

Evaluation of carboxylated poly-l-lysine for cryopreservation of Labeo rohita sperm.

Carboxylated poly-l-lysine (CPLL) is an anti-freeze agent having pronounced non-permeating yet membrane stabilizing cryoprotective capabilities. The objective was to evaluate the CPLL supplementation in extender in terms of post-thaw quality (sperm), total anti-oxidant activity (milt) and fertilization potential of cryopreserved Labeo rohita sperm. For this purpose, male brood fish reared at a fish seed hatchery, Rawal Town Islamabad, Pakistan were captured from different rearing ponds and acclimatized in hatchery ponds for 6 h. The brooder was injected with Ovaprim (0.2 mL/kg), and milt was collected after 8 h in cooled sterilized falcon tubes, maintained at 4°C and evaluated for sperm motility. The milt collected from three brooders (n = 3) was diluted in extenders viz., modified Kurokura-2 extender having 10% methanol (control); experimental extenders with CPLL supplementation at the rate of 0.5%, 1% and 1.5%. Diluted milt was filled in 0.5 mL straws, exposed to liquid nitrogen vapours and cryopreserved. Cryopreserved milt was thawed at 25°C and assessed for post-thaw sperm quality. Sperm motility, motility duration, viability, total anti-oxidant capacity and DNA integrity was significantly higher (p < 0.05) in the extender having 1.5% CPLL than control. To evaluate the fertilization rates, male and female brooders were injected with Ovaprim at 0.2 mL/Kg and 0.5 mL/Kg body weight respectively. Fresh eggs and milt were collected through abdominal stripping. Batches of 10 g of eggs from each female (n = 2) were fertilized with one straw, each from frozen sperm with KE + methanol (control), KE + methanol + 1.5% CPLL and 50 µL fresh milt (negative control). After 1.5 h of fertilization, eggs were collected from all jars and a total of 200 eggs were counted. The fertilized eggs appeared clear and transparent while unfertilized eggs looked opaque with disintegrated nuclei. Sperm fertilization rate (%) was higher (p < 0.05) in extender KE + methanol + 1.5% CPLL (78.7 ± 0.5) compared to control (KE + methanol) (52.0 ± 0.4) however, it was lower compared to that of negative control, the fresh milt (85.2 ± 0.6). In conclusion, supplementation of carboxylated poly-l-lysine (1.5%) to modified Kurokura-2 extender having 10% methanol improves post-thaw motility, motility duration, viability, DNA integrity, anti-oxidant capacity (milt) and fertilizing ability of cryopreserved L. rohita sperm.

Proteomic analysis of liver tissue reveals Aeromonas hydrophila infection mediated modulation of host metabolic pathways in Labeo rohita.

Aeromonas hydrophila (Ah) is a Gram-negative bacterium and a serious global pathogen causing Motile Aeromonas Septicaemia (MAS) in fish leading to global loss in aquaculture. Investigation of the molecular alterations of host tissues such as liver could be a powerful approach to identify mechanistic and diagnostic immune signatures of disease pathogenesis. We performed a proteomic analysis of Labeo rohita liver tissue to examine the protein dynamics in the host cells during Ah infection. The proteomic data was acquired using two strategies; discovery and targeted proteomics. Label-free quantification was performed between Control and challenged group (AH) to identify the differentially expressed proteins (DEPs). A total of 2525 proteins were identified and 157 were DEPs. DEPs include metabolic enzymes (CS, SUCLG2), antioxidative proteins, cytoskeletal proteins and immune related proteins (TLR3, CLEC4E). Pathways like lysosome pathway, apoptosis, metabolism of xenobiotics by cytochrome P450 were enriched by downregulated proteins. However, upregulated proteins majorly mapped to innate immune system, signaling of B cell receptor, proteosome pathway, ribosome, carbon metabolism and protein processing in ER. Our study would help in exploring the role of Toll-like receptors, C-type lectins and, metabolic intermediates like citrate and succinate in Ah pathogenesis to understand the Ah infection in fish. SIGNIFICANCE: Bacterial diseases such as motile aeromonas septicaemia (MAS) are among the most serious problems in aquaculture industry. Small molecules that target the metabolism of the host have recently emerged as potential treatment possibilities in infectious diseases. However, the ability to develop new therapies is hampered due to lack of knowledge about pathogenesis mechanisms and host-pathogen interactions. We examined alterations in the host proteome during MAS caused by Aeromonas hydrophila (Ah) infection, in Labeo rohita liver tissue to find cellular proteins and processes affected by Ah infection. Upregulated proteins belong to innate immune system, signaling of B cell receptor, proteosome pathway, ribosome, carbon metabolism and protein processing. Our work is an important step towards leveraging host metabolism in targeting the disease by providing a bigger picture on proteome pathology correlation during Ah infection.

A high-quality chromosome-level genome assembly of rohu carp, Labeo rohita, and its utilization in SNP-based exploration of gene flow and sex determination.

Labeo rohita (rohu) is a carp important to aquaculture in South Asia, with a production volume close to Atlantic salmon. While genetic improvements to rohu are ongoing, the genomic methods commonly used in other aquaculture improvement programs have historically been precluded in rohu, partially due to the lack of a high-quality reference genome. Here we present a high-quality de novo genome produced using a combination of next-generation sequencing technologies, resulting in a 946 Mb genome consisting of 25 chromosomes and 2,844 unplaced scaffolds. Notably, while approximately half the size of the existing genome sequence, our genome represents 97.9% of the genome size newly estimated here using flow cytometry. Sequencing from 120 individuals was used in conjunction with this genome to predict the population structure, diversity, and divergence in three major rivers (Jamuna, Padma, and Halda), in addition to infer a likely sex determination mechism in rohu. These results demonstrate the utility of the new rohu genome in modernizing some aspects of rohu genetic improvement programs.

Body composition, growth performance and enzyme activities of Labeo rohita fed different commercial fish feeds / Composição corporal, desempenho de crescimento e atividades enzimáticas de diferentes alimentos para peixes comerciais alimentados por Labeo rohita

Abstract We have evaluated the effects of different fish feeds on the body composition, growth, and enzyme activities of Labeo rohita (Rohu). In total, 240 fishes between the average weights of 24.77±2.15g were studied. The treatments were applied in a completely randomized design, with 4 treatments of 60 fishes each. Treatments consisted of four different fish feeds [Oryza (T1), AMG (T2), Aqua (T3), and Supreme (T4)]. Body composition, growth performance, and enzyme activities were evaluated. There was a significant variation in performance of fishes fed with different type of feed; as fishes having Oryza feed showed the highest weight gain, specific growth rate (SGR), and best feed conversion ratio (FCR) as compared to other groups that were considered to be significant (P ≤ 0.05). High net weight gain was obtained in T4 when compared with T2 and T3. FCR value of T4 was less than T1 but higher than T2, T3 and T2, which showed the lowest values. The specific growth rate was recorded as average in T4, but T2 led a high SGR than T3. Similarly, crude protein level and digestive enzymes activity was recorded significantly highest in fed with Oryza (T1) as compared to AMG (T2), Aqua (T3), and Supreme (T4). Water quality parameters were recorded significant in all treatments except pH and DO of treatment (T1), significantly different from other treatments. It was concluded that Rohu (Labeo rohita) could show a promising growth rate and protease enzyme activity when fed with the Oryza feed of 25% protein.

Resumo Avaliamos os efeitos de diferentes alimentos para peixes em relação à composição corporal, crescimento e atividades enzimáticas de Labeo rohita (Rohu). No total, foram estudados 240 peixes com pesos médios de 24,77 ± 2,15 g. Os tratamentos foram aplicados em delineamento inteiramente casualizado, com quatro tratamentos de 60 peixes cada. Os tratamentos consistiram em quatro alimentos diferentes para peixes: Oryza (T1), AMG (T2), Aqua (T3) e Supreme (T4). Foram avaliados a composição corporal, o desempenho de crescimento e as atividades enzimáticas. Houve uma variação significativa no desempenho dos peixes alimentados com diferentes tipos de ração. Peixes com alimentação Oryza apresentaram maior ganho de peso, taxa de crescimento específico (SGR) e melhor taxa de conversão alimentar (FCR) em comparação com outros grupos que foram considerados significativos (P ≤ 0,05). Elevado ganho de peso líquido foi obtido em T4 quando comparado com T2 e T3. O valor da FCR de T4 foi menor que T1, mas maior que T2 e T3, que apresentaram os menores valores. A taxa de crescimento específico foi registrada como média em T4, mas T2 teve uma SGR alta do que T3. Da mesma forma, o nível de proteína bruta e a atividade das enzimas digestivas foram registrados significativamente mais altos nos peixes alimentados com Oryza (T1) em comparação com AMG (T2), Aqua (T3) e Supreme (T4). Os parâmetros de qualidade da água foram registrados como significativos em todos os tratamentos, exceto pH e OD do tratamento (T1), significativamente diferente dos demais tratamentos. Concluiu-se que Rohu (Labeo rohita) pode apresentar uma taxa de crescimento promissora e atividade da enzima protease quando alimentado com Oryza de 25% de proteína.

Body composition, growth performance and enzyme activities of Labeo rohita fed different commercial fish feeds / Composição corporal, desempenho de crescimento e atividades enzimáticas de diferentes alimentos para peixes comerciais alimentados por Labeo rohita

We have evaluated the effects of different fish feeds on the body composition, growth, and enzyme activities of Labeo rohita (Rohu). In total, 240 fishes between the average weights of 24.77±2.15g were studied. The treatments were applied in a completely randomized design, with 4 treatments of 60 fishes each. Treatments consisted of four different fish feeds [Oryza (T1), AMG (T2), Aqua (T3), and Supreme (T4)]. Body composition, growth performance, and enzyme activities were evaluated. There was a significant variation in performance of fishes fed with different type of feed; as fishes having Oryza feed showed the highest weight gain, specific growth rate (SGR), and best feed conversion ratio (FCR) as compared to other groups that were considered to be significant (P ≤ 0.05). High net weight gain was obtained in T4 when compared with T2 and T3. FCR value of T4 was less than T1 but higher than T2, T3 and T2, which showed the lowest values. The specific growth rate was recorded as average in T4, but T2 led a high SGR than T3. Similarly, crude protein level and digestive enzymes activity was recorded significantly highest in fed with Oryza (T1) as compared to AMG (T2), Aqua (T3), and Supreme (T4). Water quality parameters were recorded significant in all treatments except pH and DO of treatment (T1), significantly different from other treatments. It was concluded that Rohu (Labeo rohita) could show a promising growth rate and protease enzyme activity when fed with the Oryza feed of 25% protein.

Avaliamos os efeitos de diferentes alimentos para peixes em relação à composição corporal, crescimento e atividades enzimáticas de Labeo rohita (Rohu). No total, foram estudados 240 peixes com pesos médios de 24,77 ± 2,15 g. Os tratamentos foram aplicados em delineamento inteiramente casualizado, com quatro tratamentos de 60 peixes cada. Os tratamentos consistiram em quatro alimentos diferentes para peixes: Oryza (T1), AMG (T2), Aqua (T3) e Supreme (T4). Foram avaliados a composição corporal, o desempenho de crescimento e as atividades enzimáticas. Houve uma variação significativa no desempenho dos peixes alimentados com diferentes tipos de ração. Peixes com alimentação Oryza apresentaram maior ganho de peso, taxa de crescimento específico (SGR) e melhor taxa de conversão alimentar (FCR) em comparação com outros grupos que foram considerados significativos (P ≤ 0,05). Elevado ganho de peso líquido foi obtido em T4 quando comparado com T2 e T3. O valor da FCR de T4 foi menor que T1, mas maior que T2 e T3, que apresentaram os menores valores. A taxa de crescimento específico foi registrada como média em T4, mas T2 teve uma SGR alta do que T3. Da mesma forma, o nível de proteína bruta e a atividade das enzimas digestivas foram registrados significativamente mais altos nos peixes alimentados com Oryza (T1) em comparação com AMG (T2), Aqua (T3) e Supreme (T4). Os parâmetros de qualidade da água foram registrados como significativos em todos os tratamentos, exceto pH e OD do tratamento (T1), significativamente diferente dos demais tratamentos. Concluiu-se que Rohu (Labeo rohita) pode apresentar uma taxa de crescimento promissora e atividade da enzima protease quando alimentado com Oryza de 25% de proteína.

Body composition, growth performance and enzyme activities of Labeo rohita fed different commercial fish feeds

Abstract We have evaluated the effects of different fish feeds on the body composition, growth, and enzyme activities of Labeo rohita (Rohu). In total, 240 fishes between the average weights of 24.77±2.15g were studied. The treatments were applied in a completely randomized design, with 4 treatments of 60 fishes each. Treatments consisted of four different fish feeds [Oryza (T1), AMG (T2), Aqua (T3), and Supreme (T4)]. Body composition, growth performance, and enzyme activities were evaluated. There was a significant variation in performance of fishes fed with different type of feed; as fishes having Oryza feed showed the highest weight gain, specific growth rate (SGR), and best feed conversion ratio (FCR) as compared to other groups that were considered to be significant (P 0.05). High net weight gain was obtained in T4 when compared with T2 and T3. FCR value of T4 was less than T1 but higher than T2, T3 and T2, which showed the lowest values. The specific growth rate was recorded as average in T4, but T2 led a high SGR than T3. Similarly, crude protein level and digestive enzymes activity was recorded significantly highest in fed with Oryza (T1) as compared to AMG (T2), Aqua (T3), and Supreme (T4). Water quality parameters were recorded significant in all treatments except pH and DO of treatment (T1), significantly different from other treatments. It was concluded that Rohu (Labeo rohita) could show a promising growth rate and protease enzyme activity when fed with the Oryza feed of 25% protein.

Resumo Avaliamos os efeitos de diferentes alimentos para peixes em relação à composição corporal, crescimento e atividades enzimáticas de Labeo rohita (Rohu). No total, foram estudados 240 peixes com pesos médios de 24,77 ± 2,15 g. Os tratamentos foram aplicados em delineamento inteiramente casualizado, com quatro tratamentos de 60 peixes cada. Os tratamentos consistiram em quatro alimentos diferentes para peixes: Oryza (T1), AMG (T2), Aqua (T3) e Supreme (T4). Foram avaliados a composição corporal, o desempenho de crescimento e as atividades enzimáticas. Houve uma variação significativa no desempenho dos peixes alimentados com diferentes tipos de ração. Peixes com alimentação Oryza apresentaram maior ganho de peso, taxa de crescimento específico (SGR) e melhor taxa de conversão alimentar (FCR) em comparação com outros grupos que foram considerados significativos (P 0,05). Elevado ganho de peso líquido foi obtido em T4 quando comparado com T2 e T3. O valor da FCR de T4 foi menor que T1, mas maior que T2 e T3, que apresentaram os menores valores. A taxa de crescimento específico foi registrada como média em T4, mas T2 teve uma SGR alta do que T3. Da mesma forma, o nível de proteína bruta e a atividade das enzimas digestivas foram registrados significativamente mais altos nos peixes alimentados com Oryza (T1) em comparação com AMG (T2), Aqua (T3) e Supreme (T4). Os parâmetros de qualidade da água foram registrados como significativos em todos os tratamentos, exceto pH e OD do tratamento (T1), significativamente diferente dos demais tratamentos. Concluiu-se que Rohu (Labeo rohita) pode apresentar uma taxa de crescimento promissora e atividade da enzima protease quando alimentado com Oryza de 25% de proteína.

Ecological Monitoring and Omics: A Comprehensive Comparison of Workflows for Mass Spectrometry-Based Quantitative Proteomics of Fish (Labeo rohita) Liver Tissue.

Introduction: The liver is highly sensitive to the environmental factors. Liver tissue, particularly from fish, is often used as a biological target in ecological monitoring, disease research, and stress response studies. Labeo rohita (rohu) is a fish with a significant role in the global aquaculture economy. Methods: Bottom-up proteomics relies on efficient sample preparation for performing mass spectrometric analysis of the liver tissue. Optimization of protein solubilization and digestion strategies is the key step to obtain reliable data for a successful proteomics experiment. Because the goal of extraction is to acquire the optimum protein quality and yield, the first step should be to choose an appropriate extraction method based on the type of sample. Solubilization buffers containing sodium dodecyl sulfate (SDS) or urea, and digestion methods such as filter-aided sample preparation (FASP), suspension trap (S-Trap) and in-solution are often used in proteomics but are in need of comparative evaluation with an eye to protocol optimization. Experiment: We applied two different solubilization buffers (one containing SDS, and other containing urea) and three digestion methods (FASP, S-Trap, and in-solution) to the proteomic analysis of the fish (L. rohita) liver tissue. Label-free quantification analysis was performed to analyze the similarities and differences in the results with each method. Gene ontology-based functional analysis was performed for the identified proteome across the experimental conditions to overview their protein classes, molecular functions, and biological processes. Results: SDS lysis followed by S-Trap digestion outperformed the other combinations of lysis and digestion in terms of higher protein coverage, consistency in the results and repeatability. Filter-based methods provided comparatively better results than in-solution digestion. Discussion: This protocol presents new insights on ways to optimize discovery and targeted proteomic analyses of liver tissue using the fish L. rohita as a case study. Other tissues can also be evaluated in the future drawing from the results in this study. This would help the scientific community with hypothesis-driven studies on topics ranging from basic biology to applied aquaculture research and ecological monitoring. This is particularly relevant in the current era of ecological crises and environmental pollution, where advances and optimization in research protocols can contribute to in-depth studies of ecosystems and planetary health.

Copper and cobalt doped bioactive glass-fish dermal collagen electrospun mat triggers key events of diabetic wound healing in full-thickness skin defect model.

The wounds arising out of underlying hyperglycemic conditions such as diabetic foot ulcers demand a multifunctional tissue regeneration approach owing to several deficiencies in the healing mechanisms. Herein, four different types of electrospun microfibers by combining Rohu fish skin-derived collagen (Fcol) with a bioactive glass (BAG)/ion-doped bioactive glass, namely, Fcol/BAG, Fcol/CuBAG, Fcol/CoBAG, and Fcol/CuCoBAG was developed to accelerate wound healing through stimulation of key events such as angiogenesis and ECM re-construction under diabetic conditions. SEM analysis shows the porous and microfibrous architecture, while the EDX mapping provides evidence of the incorporation of dopants inside various inorganic-organic composite mats. The viscoelastic properties of the microfibrous mats as measured by a nano-DMA test show a higher damping factor non-uniform tan-delta value. The maximum ultimate tensile strength and toughness are recorded for fish collagen with copper doped bioactive glass microfibers while the least values are demonstrated by microfibers with cobalt dopant. In vitro results demonstrate excellent cell-cell and cell-material interactions when human dermal fibroblasts (HDFs) were cultured over the microfibers for 48 h. When these mats were applied over full-thickness diabetic wounds in the rabbit model, early wound healing is attained with Fcol/CuBAG, Fcol/CoBAG, and Fcol/CuCoBAG microfibers. Notably, these microfibers-treated wounds demonstrate a significantly (p < 0.01) higher density of blood vessels by CD-31 immunostaining than control, Duoderm, and Fcol/BAG treated wounds. Mature collagen deposition and excellent ECM remodeling are also evident in wounds treated with fish collagen/ion-doped bioactive glass microfibers suggesting their positive role in diabetic wound healing.

Engineering Vascularizing Electrospun Dermal Grafts by Integrating Fish Collagen and Ion-Doped Bioactive Glass.

Utilizing bioactive molecules from organic sources in combination with inorganic materials for enhanced tissue regeneration has been a focus of recent scientific advancements. Some recent studies showed the potential of some specialized bioactive glass for healing of soft tissues; the role of Rohu (Labeo rohita) skin-derived collagen, a biopolymer in tissue regeneration and cutaneous healing, is yet to be established. So, we have fabricated four different types of electrospun mats as wound dressing materials/dermal grafts by combining locally sourced fish (Rohu) skin-derived collagen with novel composition of bioactive glass (Fcol/BAG) without and with dopants (3% and 5% Cu and Co, respectively and their binary) aimed at achieving an accelerated wound healing. FTIR and EDX mapping indicated successful integration of collagen and ion-doped bioactive glass in electrospun mats. Microfibers' architectural features and composition provided a cytocompatible and nontoxic environment conducive to adhesion, spreading, and proliferation of human dermal fibroblasts in vitro; in addition, they were hemocompatible with rabbit red blood cells. Better cutaneous wound healing in rabbits was achieved by treating with Fcol/CoBAG and Fcol/CuCoBAG microfibers with respect to improved wound closure, well-formed continuous epidermis, higher wound maturity, and regulated deposition of extracellular matrix components; mature collagen and elastin. Notably, a significantly (p < 0.01) higher density of blood vessels/positive CD 31 staining was observed in fish collagen/ion-doped bioactive glass microfibrous mat treated wounds suggesting efficient neo-vascularization during early stages of the healing process particularly attributable to copper and cobalt ions in the doped bioactive glass. Enhanced vascularizing ability of these engineered dermal composite grafts/wound dressings along with efficient remodeling of cutaneous structural components (ECM) could collectively be ascribed to bioactive properties of bioactive glass and stimulatory roles of copper, cobalt ions, and fish collagen. Our study demonstrates that a fish collagen/Cu and Co-doped bioactive glass microfibrous mat could potentially be used as a low-cost dressing material/dermal graft for augmented cutaneous wound healing.

Organ-Based Proteome and Post-Translational Modification Profiling of a Widely Cultivated Tropical Water Fish, Labeo rohita.

Proteomics has enormous applications in human and animal research. However, proteomic studies in fisheries science are quite scanty particularly for economically important species. Few proteomic studies have been carried out in model fish species, but comprehensive proteomics of aquaculture species are still scarce. This study aimed to perform a comprehensive organ-based protein profiling of important tissue samples for one of the most important aquaculture species,Labeo rohita.Deep proteomic profiling of 17 histologically normal tissues, blood plasma, and embryo provided mass-spectrometric evidence for 8498 proteins at 1% false discovery rate that make up about 26% of the total annotated protein-coding sequences in Rohu. Tissue-wise expression analysis was performed, and the presence of several biologically important proteins was also verified using a targeted proteomic approach. We identified the global post-translational modifications (PTMs) in terms of acetylation (N-terminus and lysine), methylation (N-terminus, lysine, and arginine), and phosphorylation (serine, threonine, and tyrosine) to present a comprehensive proteome resource. An interactive web-based portal has been developed for an overall landscape of protein expression across the studied tissues of Labeo rohita (www.fishprot.org). This draft proteome map of Labeo rohita would advance basic and applied research in aquaculture to meet the most critical challenge of providing food and nutritional security to an increasing world population.

Utilization of protein isolates from rohu (Labeo rohita) processing waste through incorporation into fish sausages; quality evaluation of the resultant paste and end product.

BACKGROUND: Fish processing waste could be valorized by recuperating the nutrients for human consumption. Employing a suitable process would address the pollution problems associated with its dumping and would contribute to food security. In this study, protein isolates obtained from rohu (Labeo rohita) processing waste through the pH shift process (pH 13.0) were valorized through incorporation into pangasius (Pangasius pangasius) mince sausages (0.0, 50, 100, 250, and 500 g kg-1 ). These sausages were analyzed for their proximate composition, and their physical and eating quality. RESULTS: Increasing the isolate content (11.57% to 16.14%) in sausages increased their protein content and whiteness values (P < 0.05). At 250 g kg-1 level, the cooking yield was maximum, and the expressible fluid content of paste was minimum. Moreover, adding rohu protein isolates in pangasius mince increased the expressible moisture and decreased its folding and gel strength values (P < 0.05). The gel strength values were highest for the sausages without isolates (2648.67 g cm). In contrast, in the sausages containing 250, 100, and 500 g kg-1 isolate, the values were determined to be 847.88, 731.21, and 611.06 g cm, respectively. Replacing pangasius mince with rohu protein isolates had a non-significant effect on the sensory attributes of the sausages. CONCLUSION: Replacing pangasius mince with rohu protein isolates at 250 g kg-1 resulted in superior quality sausages compared with other levels tested. Incorporating rohu protein isolates in fish sausages improved the nutritional value without significantly reducing their eating quality. The findings of this study indicate promising potential for developing nutrient-rich foods using fish protein isolates. © 2021 Society of Chemical Industry.

Efficacy of ulvan on immune response and immuno-antioxidant gene modulation in Labeo rohita against columnaris disease.

This study reports the effect of ulvan enriched diet on the influence of growth, changes in hemato-biochemical indices, improvement of antioxidant system, enhancement of innate-adaptive immunity and modification of immuno-antioxidant genes expression in Labeo rohita against Flavobacterium columnaris. The weight gain (WG) was significantly high (P > 0.05) in unchallenged normal and challenged fish fed with diets enriched with 25 and 50 mg kg-1 ulvan; the FCR was better (P > 0.05) when fed with 50 mg kg-1 enriched diet. In normal fish fed with or without ulvan supplementation was noted 100% survival rate (SR). In both groups, the red blood cell (RBC) and while blood cell (WBC) counts increased significantly (P > 0.05) when fed with 50 mg kg-1 ulvan diet whereas the hemoglobin (Hb) level increased significantly on being fed with 25 and 50 mg kg-1 ulvan diets. The SOD activity was enhanced significantly in both groups fed with any dose of ulvan diets whereas the MDA and GPx activity increased only with 25 and 50 mg kg-1 ulvan diets. The phagocytic (PC) activity significantly increased with any enriched diet and control diet groups while the respiratory burst (RB) activity increased only with 50 mg kg-1 ulvan diet. The alternate complement pathway (ACP), activity of lysozyme (Lyz), and immunoglobuline M (IgM) were better in both groups fed with 50 mg kg-1 ulvan diet. The SOD and GPx antioxidant gene expression were significantly high in both groups fed with any ulvan diet while the Nrf2 gene expression was high with 50 mg kg-1 ulvan diet. The IL-1ß, TNFα, hepcidin, Lyz, and IgM cytokines or proteins mRNA expression were significant in both groups fed with all ulvan supplement diet whereas the ß-2M expression was significant only with 50 mg kg-1 ulvan diet. The present research indicates that both L. rohita groups fed with 50 mg kg-1 ulvan diet significantly improved growth, antioxidant system, immune defense system, and immuno-antioxidant related gene expression against F. columnaris.

Effect of Green Algae Chaetomorpha antennina Extract on Growth, Modulate Immunity, and Defenses against Edwardsiella tarda Infection in Labeo rohita.

The current study focused on assessing the outcome of methanol extract of Chaetomorpha antennina (MECA) on the growth performance and immune modulation in both specific and non-specific immune responses through the assessment of neutrophil, serum lysozyme, serum myeloperoxidase, antiprotease, ceruloplasmin, reactive oxygen species (ROS), and reactive nitrogen species (RNS) activity in Labeo rohita (rohu) at 28 days post treatment along with assessment of the disease resistance capacity against Edwardsiella tarda at 30days post immunization. Fishes (n = 144; average weight 50.0 ± 0.23 g) were evenly divided into four treatments, with 12 fishes per tank in triplicates. The MECA was injected intraperitoneally in the fishes at different doses as 0, 25, 75, and 150 mg/kg of the body weight. The results demonstrated that fish treated with MECA have an increased body weight, specific growth rate, and feed conversion ratio (p < 0.05) with respect to the control group. Results suggested that the MECA inclusion can significantly enhance (p < 0.05) the levels of serum lysozyme, neutrophil function, serum antiprotease activity, cellular RNS, and ROS production. Exposure to MECA of 75 mg/kg showed a significantly higher survival percentage against E. tarda disease infection. These results indicate MECA as a stimulant of immunity in L. rohita against E. tarda. The results suggested that MECA is a potent immunostimulant in finfish aquaculture and can offer higher economic welfare.

Effect of organic acid on recovery yields and characteristics of rohu (Labeo rohita) protein isolates extracted using pH shift processing.

BACKGROUND: Proteins recovery using hydrochloric acid (HCl) in acid-aided solubilization could cause greater loss in its functionality over alkali-aided solubilization. Moreover, using HCl in edible foods is also a health concern. Replacing HCl with organic acids for acid-aided solubilization could address these problems. The aim was to study the effect of organic acid (glacial acetic acid) as a replacement for HCl during pH shift processing on the characteristics and functionality of rohu (Labeo rohita) protein isolates. Rohu proteins were obtained by solubilizing at pH 3.0 and pH 11.0 using glacial acetic acid and sodium hydroxide (10 mol L-1 ). RESULTS: Results showed that solubilization at pH 11.0 gave higher protein yields (766.8 ± 2.4 g kg-1 ) compared to solubilization at pH 3.0 (735.7 ± 7.1 g kg-1 ) (P < 0.05). Isolates from acid-aided solubilization had higher whiteness and total pigment content over isolates obtained by alkali-aided solubilization. Rohu isolates recovered by alkaline solubilization showed higher water and oil holding capacity, gel strength, folding scores, foaming and emulsion capacity than acid processed isolates (P < 0.05). Solubilization of rohu proteins using glacial acetic acid produced isolates with low breaking force (149.0 g), low storage modulus (G') values and low folding test score (1.0) over the alkaline isolates (P < 0.05). CONCLUSION: Results indicated that, recovering rohu proteins using organic acid (glacial acetic acid) could produce isolates with poor functional properties, while using the organic acid to precipitate the proteins solubilized by alkali-aided processing could produce proteins with better yields and functionality. © 2019 Society of Chemical Industry.