Healthy Fish Research Articles

Molecular cloning and functional analyses of C–C motif chemokine ligand 3 (CCL3) in mandarin fish Siniperca chuatsi

Chemokines are critical molecules involved in immune reaction and immune system homeostasis, and some chemokines play a role in antiviral immunity. It is not known if the C–C motif chemokine ligand 3 (CCL3), a member of the CC chemokine family, possesses antiviral properties in fish. In this study, a ccl3 was cloned from the mandarin fish (Siniperca chuatsi), and it has an open reading frame (ORF) of 276 base pairs, which are predicted to encode a 91-amino acid peptide. Mandarin fish CCL3 revealed conserved sequence features with four cysteine residues and closely relationships with the CCL3s from other vertebrates based on the sequence alignment and phylogenetic analysis. The transcripts of ccl3 were notably enriched in immune-related organs, such as spleen and gills in healthy mandarin fish, and the ccl3 was induced in the isolated mandarin fish brain (MFB) cells following infection with infectious spleen and kidney necrosis virus (ISKNV). Moreover, in MFB cells, overexpression of CCL3 induced immune factors, such as IL1β, TNFα, MX, IRF1 and IFNh, and exhibited antiviral activity against ISKNV. This study sheds light on the immune role of CCL3 in immune response of mandarin fish, and its antiviral defense mechanism is of interest for further investigation.

Effects of dietary nanoliposome-coated astaxanthin on haematological parameters, immune responses and the antioxidant status of rainbow trout (Oncorhynchus mykiss).

Astaxanthin is the most prevalent carotenoid in the marine environment and is widely used as an additive in formulated aquafeeds. A 60-day feeding trial was conducted to consider the effect of dietary nanoliposome-coated astaxanthin (NA) on haematological parameters, serum antioxidant activities and immune responses of rainbow trout, Oncorhynchus mykiss. A total of 450 healthy fish weighing 31.00±2.09g were randomly assigned in triplicate (30 fish per replicate) to 5 dietary treatments: 0 (control), 25.00, 50.00, 75.00, and 100.00mgkg-1 NA. Fish fed the diet supplemented with 50.00mgkg-1 NA exhibited the highest values of red blood cells, white blood cells, haemoglobin and haematocrit of 1.64±0.01×106mm-3, 5.54±0.21×103mm-3, 8.73±0.24gdL-1 and 46.67%±0.88%, respectively, which were significantly higher than those fed the basal diet (p<0.05). The lowest and highest percentages of lymphocytes (67.67%±0.33%) and neutrophils (27.33%±1.20%) were also obtained in fish fed 50.00mgkg-1 NA compared to those fed the basal diet (p<0.05). Fish receiving diet supplemented with 50.00mgkg-1 NA revealed the highest serum activity in superoxide dismutase, catalase, glutathione peroxidase, lysozyme and alternative complement and the lowest level of total cholesterol, cortisol, aspartate aminotransferase and alanine aminotransferase than fish receiving the basal diet (p<0.05). Serum immunoglobulin (Ig) and ACH50 contents significantly increased with increasing dietary NA supplementation to the highest values of 43.17±1.46 and 293.33±2.03UmL-1, respectively, in fish fed diet supplemented with 50mgkg-1 NA (p<0.05). Supplementation of NA in rainbow trout diet at 50mgkg-1 exhibited a positive effect on haematological parameters, antioxidant capacity and immune responses. Administration of such dosage can enhance rainbow trout immune responses against unfavourable or stressful conditions, for example disease outbreaks, hypoxic condition, thermal stress and sudden osmotic fluctuations, which usually happen in an intensive culture system.

Identification and characterization of STAT family in silver pomfret (Pampus argenteus) involved in different exogenous stresses

Members of the Signal Transducer and Activator of Transcription (STAT) family function pivotally as transcriptional activators integral to the modulation of inflammatory responses. The aquaculture of silver pomfret is frequently compromised by the imposition of exogenous stressors, which include thermal fluctuations, notably low-temperatures, diminished oxygen levels, and the onslaught of bacterial pathogens. Notwithstanding the critical impact of these stressors, the scientific literature presents a notable gap in our understanding of the STAT pathway's role in the silver pomfret's adaptive response mechanisms. To address this lacuna, we identified stat genes in the silver pomfret—denominated as Pastat1, Pastat2, Pastat3, Pastat4, and Pastat5—through a thorough and systematic bioinformatics analysis. Further scrutiny of the gene configurations and constituent motifs has elucidated that STAT proteins possess analogous structural frameworks and exhibit significant evolutionary preservation. Subsequently, the expression patterns of five stat genes were verified by RT-qPCR in twelve different tissues and four growth periods in healthy fish, showing that the expression of Pastat genes was temporally and spatially specific, with most of the stat genes expressed at higher levels in the spleen, following muscle, gill, and liver. Transcriptomic analysis of exposure to exogenous stressors, specifically formaldehyde and low-temperature conditions, elucidated that Pastat1 and Pastat2 genes exhibited a heightened sensitivity to these environmental challenges. RT-qPCR assays demonstrated a marked alteration in the expression profiles of jak1 and Pastat gene suites in PaS upon prolonged bacterial infection subsequent to these exogenous insults. Moreover, the gene expression of the downstream effectors involved in innate immunity and apoptosis displayed marked deviations. This study additionally elucidated the Pastat gene family's role in modulating the innate immune response and apoptotic regulation within the silver pomfret during exogenous stressors and subsequent pathogenic incursions.

Development and validation of a flow cytometry method to examine circulating leukocyte subpopulations in barramundi (Lates calcarifer)

Bacterial infections are a major contributor to losses in aquaculture production. Diagnosis of these bacterial diseases is based on sacrificing fish and isolating the infectious agent from the internal organs. Infections lead to an inflammatory response in the host that is characterized by changes in leukocyte profile and activity. We hypothesized that the profile of circulating peripheral blood leukocytes and markers of their activity, such as reactive oxygen species production, would change following an event of bacterial infection in fish. We therefore developed a flow cytometry-based methodology for analyzing blood leukocytes in barramundi (Lates calcarifer), then used it to compare healthy fish to those infected with a common pathogenic bacterium, Vibrio harveyi. The initial step of the analysis relied on the effective separation of erythrocytes and leukocytes, and two methods were tested: one based on hypotonic lysis of red blood cells, and the other involving stain-based differentiation using carboxyfluorescein succinimidyl ester (CFSE). Fluorescence-activated cell sorting analysis revealed significant differences in leukocyte parameters of infected and control fish using the stain-based approach vs. hypotonic lysis. Changes in blood leukocytes included an increase in the proportion of monocytes and decrease in that of lymphocytes in infected fish. Furthermore, cell size and granularity of the different leukocyte populations changed over time in infected fish as compared to controls. Monitoring peripheral blood leukocytes provides a non-lethal analysis of inflammation that allows repeated sampling of individual fish over time. This is beneficial given the high variability between individual fish. The use of CFSE-staining for leukocyte isolation proved to be advantageous over a method based on lysis of erythrocytes.

Evaluation of different probiotics on growth, body composition, antioxidant capacity, and histoarchitecture of Mugil capito

We investigated the dietary effects of the single application of Saccharomyces cerevisiae, Lactobacillus bulgaricus, and their combination on growth, proximate composition of whole fish body, antioxidant defense, and histoarchitecture of hapa-reared Mugil capito. Healthy fish (Fish weighed = 10.30 ± 0.10 g at first) were randomly allocated into 4 equal groups, each with three replicates. These groups were designed as follows: (1) a group fed a basal diet without probiotics (control), (2) a group fed a diet containing S. cerevisiae (4 g/kg diet), (3) a group fed a diet containing L. bulgaricus (2 g/kg diet), and (4) the last group fed a diet containing a combination of both, all for a duration of 60 days. Probiotic-treated groups showed significantly better growth and nutrition utilization than the control group. Significant differences were observed in the crude fat and crude protein contents among the groups, with the combination group exhibiting the highest levels. However, there were no significant variations in ash content across all groups. The highest hepatic antioxidant capacity (superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) enzyme activities) was observed in the combination group. Thiobarbituric acid reactive substance (TBARS) concentrations were decreased significantly in all probiotic groups, suggesting improved oxidative stress resilience in these groups. The histomorphological analysis of the hepatopancreatic tissues revealed well-arranged parenchyma, increased glycogen storage, and melanomacrophage centers in probiotic-treated groups, particularly the combined probiotics group. Furthermore, the probiotic supplementation improved the histoarchitecture of the intestinal villi compared to the control group. To put it briefly, combined dietary administration of these probiotics improved growth, body composition, antioxidant defenses, and hepatic and intestinal health in hapa-reared M. capito, highlighting their promising role in promoting welfare and productivity.

Etiology of Ulcerative Dermal Necrosis (UDN) in Brown Trout (Salmo trutta Morpha trutta)-Preliminary Results.

Every year, ulcerative dermal necrosis (UDN) affects salmonids that spend most of their lives in the sea during their migration to the rivers of northern Poland to spawn. The clinical form of the disease manifests itself in ulcerative skin lesions, which lead to significant weakening of the fish and, in most cases, result in their death. This study was carried out on samples taken from sea trout in the Słupia River in northern Poland. In order to identify the pathogen, experiments on the transmission of the disease were carried out, and additional histopathological, microbiological and electron microscopic examinations were performed. As a result of these studies, it was possible to experimentally transfer the disease from sick to healthy fish. The results indicate a complex etiology of the disease (lack of a clearly defined pathogen), in which the change in the environment from salty to freshwater triggers the related changes in skin physiology, which are the main causes of increased susceptibility to the development of the disease.

Clinical symptoms and histopathological changes in coho salmon affected by the erythrocytic inclusion body syndrome (EIBS) are caused by the infection of piscine orthoreovirus 2 (PRV-2).

The relationship of histopathological changes and the infection of Piscine orthoreovirus 2 (PRV-2) was investigated in coho salmon that were suffering from the erythrocytic inclusion body syndrome (EIBS). Immunohistochemical observations revealed abundant σ1 protein of PRV-2 in the spongy layer of the ventricle of the heart, where severe myocarditis was observed. In the spleen, the virus protein was detected in many erythrocytes, some of which were spherical-shaped and apparently dead. The number of erythrocytes was decreased in the spleen compared to the apparently healthy fish. The virus protein was also detected in some erythrocytes in blood vessels. The viral protein was often detected in many macrophages ingesting erythrocytes or dead cell debris in the spleen or in the kidney sinusoids. Large amounts of the viral genomic segment L2 were also detected in these organs by RT-qPCR. Many necrotic foci were found in the liver, although the virus protein was not detected in the hepatocytes. These results suggest that the primary targets of PRV-2 are myocardial cells and erythrocytes and that clinical symptoms such as anaemia or jaundice and histopathological changes such as myocarditis in EIBS-affected coho salmon are caused by PRV-2 infection.

Role of Vitamin E Supplementation on the Reproductive and Growth Performance, Hormonal Profile and Biochemical Parameters of Female Hybrid Red Tilapia

AbstractThe present study aimed to evaluate the effect of vitamin E (VE) supplementation on the reproductive and growth performance, hormonal profile, and biochemical parameters of female hybrid red tilapia before spawning season. Seventy-two female hybrid red tilapia were caught with an average body-weight 272.56 ± 34.84g and an average total length 24.3 ± 2.5 cm. Healthy fish were distributed equally into 4 treatments supplemented with (0, 50, 100, 200 mg/kg) of VE given for 8 weeks. VE significantly improved weight gain, length gain, specific growth rate, average daily gain, feed conversion ratio, and protein efficiency ratio. 50 mg of VE/kg diet could improve the viscerosomatic index, hepatosomatic index, and gonadosomatic index. Also, it could improve the parameters of biochemical (aspartate aminotransferase, alanine transaminase, total protein, albumin, globulin, triglyceride, and alkaline phosphatase) and haematology (total erythrocytic and leucocytic count, hematocrit and hemoglobin) and increase FSH, LH, E2 and, progesterone concentrations significantly. The conclusion revealed that the addition of 50 mg of VE /kg diet has a beneficial impact on reproductive and growth performance, hormonal profile, and biochemical parameters of female hybrid red tilapia. So, it is advisable for adding 50 mg/kg of vitamin E to the fish diet before the spawning season (pre-spawning).

Cloning and expression analysis of Janus activated kinase family genes from spotted seabass (Lateolabrax maculatus)

Janus kinases (JAKs) are important components of the JAK-STAT signaling pathway and play vital roles in innate immunity, autoimmune diseases, and inflammation. However, information about JAKs remains largely unknown in the spotted seabass, a fish species of Perciformes with great commercial value in the aquaculture industry. The aims of this study are to obtain the complete cDNA sequences of JAKs (JAK1, JAK2A, JAK2B, JAK3 and TYK2) from spotted seabass and to investigate their roles upon stimulation with lipopolysaccharides (LPS) and Edwardsiella tarda, using RT-PCR, PCR and qRT-PCR methods. All five JAK genes from the spotted seabass, each encode more than 1100 amino acids residues. JAK1 and JAK3 consist of 24 exons and 23 introns, whereas JAK2A, JAK2B and TYK2 consist of 23 exons and 22 introns. Furthermore, these five spotted seabass JAKs share high sequence identities with those of other fish species in protein domain analysis, synteny analysis, and phylogenetic analysis. Moreover, these five JAK genes were ubiquitously expressed in all tissues examined from healthy fish, and inducible expressions of JAKs were observed in the intestine, gill, head kidney, and spleen following LPS treatment or E. tarda infection. These findings indicate that all these JAK genes are involved in the antibacterial immunity of the spotted seabass and provide a basis for further understanding the mechanism of JAKs antibacterial response in the spotted sea bass.

Application Of Lights in The Nursery of Snubnose Pompano Trachinotus blochii (Lacepède, 1801) In Ponds as An Effort to Increase Feed Cost Efficiency

The snubnose pompano (Trachinotus blochii), which is an introduced fish, is currently in increasing demand, both for the domestic market (Jakarta and other big cities) and international markets such as Singapore, Taiwan, Hong Kong, and China. The snubnose pompano can be cultivated in ponds because it is tolerant to changes in salinity (5-40 ppt). In this study, the use of lights installed in cages used to maintain the snubnose pompano nursery will be responded to natural food that is positively phototactic (zooplankton, shrimp, and small fish species) which will gather as feed at night. So, it is hoped that the use of these organisms as an alternative feed source can increase feed efficiency. The urgency of this research is to determine efforts to save on the use of artificial feed and to suppress parasite attacks. A t-test was carried out to compare the averages of two different samples, on the growth rate and health of snubnose pompano fish. Research using lights resulted in optimum growth of snubnose pompano with 100% survival rate, feed conversion ratio (FCR) value of 0.9 and produced snubnose pompano size 80–100 g/head within two months as well as healthy fish seeds. With indicators of parasitic attack that are lower than the treatment without light with a prevalence value of 25% (with lights); range of 25%-75% (without lights) and intensity 6-8 (with lights); range 11-35.67 (without light). The snubnose pompano seeds are always actively moving when the light is on, the use of the light can effectively catch organisms such as small shrimp or small crustaceans so that the snubnose pompano seeds can be used to be digested as an alternative food (as observed in the stomach contents and intestines of the snubnose pompano). The light also indicates that parasitic attacks on the snubnose pompano are less common.

Bacillus sp. as potential probiotics for use in tilapia fish farming aquaculture – a review

Abstract Aquaculture is a crucial and rapidly expanding industry in global food production. Fisheries are, also an essential socio-economic activity, providing abundant resources and remarkable prospects. However, due to the deteriorating ecological environment, aquatic animals are often exposed to traumatic conditions and are susceptible to bacterial infections that pose significant challenges for aquaculture production. The indiscriminate use of antibiotics in the past has led to the emergence of multidrug-resistant pathogens and sudden outbreaks of infectious diseases, resulting in serious economic losses. Moreover, the use of expensive chemotherapeutic drugs and antibiotics has negative impacts on aquatic environments. Therefore, it is increasingly important to adopt alternative natural agents, such as probiotics and their metabolites, to enhance healthy fish production. Probiotics are microorganisms that have numerous beneficial effects on their hosts. They are environmentally friendly, nontoxic, and cost-effective. This review specifically focuses on the use of Bacillus sp. as probiotics to promote healthy tilapia production in the aquatic sector, while also examining their interactions with the immune system and gut microbiota. The information presented in this review can guide future research and promote effective and healthy tilapia culture production.

Effects of Ultraviolet Radiation (UVR) on Some Stages of &amp;lt;i&amp;gt;Clarias gariespinus&amp;lt;/i&amp;gt; (Catfish) Growth

UVR is a stressor that affect ecological and social systems. It has been noted that UVR presents numerous difficulties for aquatic and human worldwide. It&amp;apos;s critical to understand how UVR affects &amp;lt;i&amp;gt;Clarias gariespinus&amp;lt;/i&amp;gt; in order to promote healthy fish growth. This study determined how UVR affected catfish. 172 catfish samples were divided into four groups: UV-A, UV-B, UV-C, and controls. The control group was not exposed, whereas the other groups were exposed to UV-A, UV-B, and UV-C, respectively. The exposure period was 131 days, from 8:00 am to 5:00 pm daily. The result on color change shows that UV-C causes a change in color from dark to pink at the fingerling stage and UV-A causes a change in color from dark to slightly pink at the jumbo size, while no color change was observed in other samples. The result on growth rate indicates that the UV-B sample grew faster throughout the period of study, with the highest growth rates of 18.4, 16.2, 14.1, and 8.6 cm for the UV-B, UV-C, control, and UV-A samples, respectively. The result on the mortality rate of the samples shows that the control sample recorded the highest death rate (23) at the fingerling stage, followed by the UV-A (22), UV-C (19), and UV-B (12) samples. The result depicts that UV-B is capable of a rapid increase in the weight, growth, and life span of catfish; hence, exposure of catfish to UV-B can be adopted by fish farmers to improve the healthy fish growth of their farm.

New sight in arginine-improved flesh quality: Role of MRFs, cyclins, and WNT signaling in grass carp (Ctenopharyngodon idellus)

In this study, we investigated the functional roles and mechanisms of arginine in improving the muscle quality of sub-adult grass carp. We divided 450 healthy fish (661.60 ± 1.00 g) into six groups and fed them separate diets with increasing levels of arginine (3.51, 7.01, 10.51, 14.01, 17.51, and 21.01 g/kg) for 63 days. The results showed that optimal dietary arginine levels promoted grass carp growth and improved muscle lipid content, shear force (hardness), pH, hydrolyzed amino acid contents, and the chemical score of essential amino acids (protein quality). Suitable dietary arginine levels also decreased the proportion of hypertrophic fibers and increased the proportion of hyperplastic fibers. Furthermore, appropriate arginine supplementation increased the expression levels of MyoD, MyoG, Myf5, MRF4, MyHC, cyclin D1, cyclin B, cyclin E, WNT1, WNT3a, and β-catenin and decreased the expression levels of MSTN and GSK3β. In conclusion, appropriate arginine levels in the diet (10.51–14.01 g/kg) may promote the growth and muscle quality of grass carp while increasing the proportion of hyperplasia fibers and decreasing the proportion of hypertrophic fibers probably by regulating the WNT/β-catenin pathway to increase the expression levels of MRFs and cyclins. This study offers novel insights into the regulatory role of arginine in muscle fiber growth, which can facilitate the improvement of grass carp muscle quality.

Identification, distribution, bactericidal and immunoregulatory activities of NK-lysins in grass carp (Ctenopharyngodon idella)

NK-lysins, which are antimicrobial peptide mainly produced by cytotoxic T lymphocytes and natural killer cells, play an important role in immune defense against infection. In this study, using CiNkla gene as template, 2 highly homologs CiNklb and CiNklc were further identified in grass carp. The three NK-lysin transcripts were ubiquitously distributed in various tissues, and CiNkla and CiNklb have the highest transcripts in spleen while CiNklc has the highest transcript in gill. Upon challenge with Aeromonas hydrophila, the transcripts of CiNkla and CiNklb were significantly increased in intestine, whereas transcript of CiNklc increased in liver. The concentration of steady-state CiNkla protein was highest in spleen in healthy fish, while upon challenge with A. hydrophila, highest level of CiNkla protein was detected in head kidney at 6 h post infection (hpi), and in liver at either 24 hpi or 48 hpi. The recombinant CiNkla and CiNklb purified from Pichia pastoris show direct bactericidal activity against both Gram-negative and Gram-positive bacterial pathogens. The A. hydrophila treated with CiNkla showed morphological change, release of intracellular contents and genome DNA degradation. Supplementation of CiNkla or CiNklb to cultured head kidney derived macrophages (HKDMs) of grass carp changes the transcription of inflammatory factors and chemokines. In vivo stimulation of fish with recombinant CiNkla or CiNklb confirmed both molecules could increase transcription of chemokines and cytokines in tissues, and liver was sensitive to stimulation and induced into an inflammation condition at earlier timepoints. Our results indicate that transcription and expression of NK-lysins are modulated by bacterial infection, and CiNkla and CiNklb could modulate host immune defense.

Behavioral and neurophysiological effects of buspirone in healthy and depression-like state juvenile salmon.

A proportion of farmed salmon in seawater show a behaviorally inhibited, growth stunted profile known as a depression-like state (DLS). These DLS fish are characterized by chronically elevated serotonergic signaling and blood plasma cortisol levels and the inability to react further to acute stress, which is suggestive of chronic stress. In this study, we characterize the neuroendocrine profile of growth stunted freshwater parr and confirm that they show a DLS-like neuroendocrine profile with a blunted cortisol response and no serotonergic increase in response to acute stress. Furthermore, we attempted to reverse this DLS-like profile through pharmacological manipulation of the serotonin (5-HT) system with buspirone, an anxiolytic medication that acts as a serotonin receptor agonist (i.e., decreases serotonergic signaling). We found that while buspirone decreases anxiolytic-type behavior in healthy fish, no quantifiable behavioral change was found in DLS-like fish. However, there was a physiological effect of diminished basal serotonergic signaling. This suggests that at the physiological level, buspirone appears to reverse the neuroendocrine DLS profile. With a deeper understanding of what causes DLS profiles and growth stunting in juvenile fish, steps can be taken in terms of husbandry to prevent repeated stressors and the formation of the DLS profile, potentially reducing losses in aquaculture due to chronic stress.

Use of Ergonomics and Analytical Hierarchy Process Model for Selection of Cities of Small Medium Enterprises

Fish Smoking is a traditional method that has long been used in fisheries. The use of fish smoking devices can have a negative impact on human health which results in the entry of smoke into the eyes, lungs, and skin. In order not to be harmful to human health, ergonomic and healthy fish smoking devices are used. This study analyzes the prioritization of Small Medium Enterprises (SMEs) groups in the use of fish smoking devices to reduce the risk of health problems. The approach method used in this analysis uses the Ergonomics and Analytical Hierarchy Process (AHP) method to determine the priorities of SMEs groups in the use of fish smoking devices. The criteria in this analysis are Ergonomics (Safe, Comfortable, Healthy), Health, Education Level, Community Participation, Motivation, Habits, and Resistance. The results of the analysis found that priority choices in percentage include E, A, B, D, and C regencies. It is hoped that with the holding of Diffusion Technology Results of Innovation Products (DTHPI), the use of modern fish smoking devices that are more ergonomic, healthy, and practical can help improve the welfare of SMEs.

Detection of Freshness of Fish using Machine Learning Techniques on Vyas Municipality, Nepal

The historical narrative of the fish trade is well-document in various sources. However, the concerning prevalence of fish traders vending spoiled fish poses a significant threat to human health, prompting specific research inquiries. The study aimed to address key questions: What quality of healthy fish do traders sell? How effective are their fish storage methods? What's the duration between fish purchase and consumer access? The study objectives were devised to uncover a actual condition of the fish on sale, assess storage practices, and determine the selling timeline. To achieve these aims, the study employed the EfficientNetB1 machine learning model, chosen for its simplicity and high accuracy. Five fish shops and traders from wards 1,2,3 and 4 in Damauli, the primary city of Vyas Municipality in Nepal, were selected for investigation. Results from five main city shops in Damauli revealed that only 26% of the fish were deemed healthy, while a concerning 74% were identified as rotten. Similarly, within the sample, 44% of the fish were healthy, while 56% were spoiled. This study unveiled that fish were being sold even up to 15 days post-purchase, employing ice packs, refrigeration, and potentially chemicals for storage. These findings highlight the urgent need for ongoing monitoring by relevant stakeholders and local government entities to address this issue effectively.

The efficiency of zinc sulfate immersion bath on improved wound healing via promoting antioxidant activity, gene expression biomarkers, and skin re-epithelization in a common carp-induced wound model

The experiment was designed to examine the influence of employing three doses of ZnSO4 on the wound healing process in partially scaled common carp. A total of 240 healthy common carp fish (52.3 ± 0.9 g) were randomly allocated into four equal groups in triplicate (20 each). The first group left without any zinc sulfate treatment and served as a control group, while the second group through the fourth group were immersed in a zinc sulfate bath at a dose of 2.09, 1.05, and 0.53 mg/L corresponding to 1/5, 1/10, and 1/20 of 96 h LC50 of Zn, (Zn/5, Zn/10, and Zn/20, respectively). After wound induction, tissue specimens were collected within three different intervals (6 h, 24 h, 72 h, and 14 days). The results indicated that the Zn/5 fish group induced doubled folding increments in the expression of transforming growth factor (TGF)‐β1 after 6 h compared to other groups, whereas collagen type I alpha 1 (COL1α1) and metallothionein (Met) genes exhibited a triple folding increment compared to Zn/10 and a fivefold increase compared to control after two days of wound induction. Moreover, vascular endothelial growth factor (VEGF)‐A and fibroblast growth factor (FGF)‐7 genes showed a dose-dependent manner of expression at all examined points after wound induction. Also, all estimated antioxidant biomarker (superoxide-dismutase, SOD; catalase, CAT; glutathione, GSH; and malonaldehyde, MDA) activities were boosted in the Zn/5 group till three days of wound induction compared to all groups. In addition, the reepithelization score and histological alteration results revealed clear improvement in the Zn/5 group, as most muscle fibers appeared regular, straight, and parallel arranged. In contrast, other groups exhibited a detectable limited area of disrupted muscle fibers. Finally, it could be concluded that the ZnSO4 immersion bath at 1/5 of the calculated LC50 effectively enhanced the healing process and skin reepithelization.

Automated Hematological Approach and Protein Electrophoretic Pattern in Tilapia (Oreochromis niloticus): An Innovative and Experimental Model for Aquaculture

This study aimed to assess the usefulness of two innovative automated methods (automated blood count counters and flow cytometry) for hematological investigation in Tilapia to make a contribution to the clinical diagnostics of this farmed species. Moreover, serum total proteins and their electrophoretic fractions (prealbumin, albumin, α-, β-, and γ-fraction), as health condition indicators, were assessed. The analysis of serum total proteins and electrophoretic fraction showed a normal and typical electrophoretic pattern of healthy fish (serum total proteins, 3.70 ± 0.62 g/dL; prealbumin, 0.44 ± 0.20 g/dL; albumin, 1.17 ± 0.66 g/dL; α-fraction, 1.49 ± 0.64 g/dL; β-fraction, 0.32 ± 0.16 g/dL; and γ-fraction, 0.29 ± 0.13 g/dL). The relationships between the values of red blood cells (RBCs), white blood cells (WBCs), and thrombocytes (TCs) obtained with the two automated methods were determined using Pearson correlation analysis. The results showed a significant positive correlation between automatic blood cell counting and flow cytometry analysis for RBCs (r = 0.97, p &lt; 0.0001) and WBCs (r = 0.91, p &lt; 0.0001), whereas no correlation was found for TCs (r = −0.11, p = 0.66). The preliminary results gathered in this study seem to highlight the usefulness of the new analytical techniques herein investigated in tilapia, suggesting their application in the hematological investigation of farmed fish species and their usefulness for monitoring the health and well-being of fish reared in aquaculture.

Assessing the influence of preconception diet on male fertility: a systematic scoping review.

The last decade has seen increased research on the relationship between diet and male fertility, but there are no clearly defined nutritional recommendations for men in the preconception period to support clinical fertility outcomes. The purpose of this scoping review is to examine the extent and range of research undertaken to evaluate the effect(s) of diet in the preconception period on male clinical fertility and reproductive outcomes. Four electronic databases (MEDLINE and EMBASE via Ovid, CAB Direct, and CINAHL via EBSCO) were searched from inception to July 2023 for randomized controlled trials (RCTs) and observational studies (prospective/retrospective, case-control, and cross-sectional). Intervention studies in male participants or couples aiming to achieve dietary or nutritional change, or non-intervention studies examining dietary or nutritional components (whole diets, dietary patterns, food groups or individual foods) in the preconception period were included. Controls were defined as any comparison group for RCTs, and any/no comparison for observational studies. Primary outcomes of interest included the effect(s) of male preconception diet on clinical outcomes such as conception (natural or via ART), pregnancy rates and live birth rates. Secondary outcomes included time to conception and sperm parameters. A total of 37 studies were eligible, including one RCT and 36 observational studies (prospective, cross-sectional, and case-control studies; four studies in non-ART populations) published between 2008 and 2023. Eight reported clinical outcomes, 26 reported on secondary outcomes, and three reported on both. The RCT did not assess clinical outcomes but found that tomato juice may benefit sperm motility. In observational studies, some evidence suggested that increasing fish or reducing sugar-sweetened beverages, processed meat or total fat may improve fecundability. Evidence for other clinical outcomes, such as pregnancy rates or live birth rates, showed no relationship with cereals, soy and dairy, and inconsistent relationships with consuming red meat or a 'healthy diet' pattern. For improved sperm parameters, limited evidence supported increasing fish, fats/fatty acids, carbohydrates and dairy, and reducing processed meat, while the evidence for fruits, vegetables, cereals, legumes, eggs, red meat and protein was inconsistent. Healthy diet patterns in general were shown to improve sperm health. Specific dietary recommendations for improving male fertility are precluded by the lack of reporting on clinical pregnancy outcomes, heterogeneity of the available literature and the paucity of RCTs to determine causation or to rule out reverse causation. There may be some benefit from increasing fish, adopting a healthy dietary pattern, and reducing consumption of sugar-sweetened beverages and processed meat, but it is unclear whether these benefits extend beyond sperm parameters to improve clinical fertility. More studies exploring whole diets rather than singular foods or nutritional components in the context of male fertility are encouraged, particularly by means of RCTs where feasible. Further assessment of core fertility outcomes is warranted and requires careful planning in high-quality prospective studies and RCTs. These studies can lay the groundwork for targeted dietary guidelines and enhance the prospects of successful fertility outcomes for men in the preconception period. Systematic search of preconception diet suggests that increasing fish and reducing sugary drinks, processed meats and total fat may improve male fertility, while consuming healthy diets, fish, fats/fatty acids, carbohydrates and dairy and reducing processed meat can improve sperm health.