Acquisition and Analysis Methods for Hi-C Data from Medaka Early Embryos.

The three-dimensional (3D) structure of the genome undergoes dynamic changes during the developmental process. While Hi-C is a technique that enables the acquisition of genome 3D structure data across various species and cell types, existing Hi-C analysis programs may face challenges in detecting and comparing structures effectively depending on the characteristics of the genome or cell type. Here, we describe a method for acquiring Hi-C data from medaka early embryos and quantifying the structural changes during the developmental process.

Metabolic and protein expression responses of Shewanella baltica in golden pomfret broths to slightly acidic electrolysed water.

Shewanella baltica is a specific spoilage organism of golden pomfret. This study aims to explore the antibacterial mechanism of slightly acidic electrolysed water (SAEW) against S. baltica (strains ABa4, ABe2 and BBe1) in golden pomfret broths by metabolomics, proteomics and bioinformatics analyses. S. baltica was decreased by at least 3.94 log CFU/mL after SAEW treatment, and strain ABa4 had the highest resistance. Under SAEW stress, amino acids and organic acids in S. baltica decreased, and nucleotide related compounds degraded. Furthermore, 100 differentially expressed proteins (DEPs) were identified. Most DEPs of strains ABe2 and BBe1 were down-regulated, while some DEPs of strain ABa4 were up-regulated, especially those oxidative stress related proteins. These results suggest that the modes of SAEW against S. baltica can be traced to the inhibition of amino acid, carbon, nucleotide and sulphur metabolisms, and the loss of functional proteins for temperature regulation, translation, motility and protein folding.

Development and antimicrobial activity of composite edible films of chitosan and nisin incorporated with perilla essential oil-glycerol monolaurate emulsions.

Aquatic products are highly susceptible to spoilage, and preparing composite edible film with essential oil is an effective solution. In this study, composite edible films were prepared using perilla essential oil (PEO)-glycerol monolaurate emulsions incorporated with chitosan and nisin, and the film formulation was optimized by response surface methodology. These films were applied to ready-to-eat fish balls and evaluated over a period of 12 days. The films with the highest inhibition rate against Staphylococcus aureus were acquired using a polymer composition of 6 µL/mL PEO, 18.4 µg/mL glycerol monolaurate, 14.2 mg/mL chitosan, and 11.0 µg/mL nisin. The fish balls coated with the optimal edible film showed minimal changes in appearance during storage and significantly reduced total bacterial counts and total volatile basic nitrogen compared to the control groups. This work indicated that the composite edible films containing essential oils possess ideal properties as antimicrobial packaging materials for aquatic foods.

Molecularly imprinted polymers-coated magnetic covalent organic frameworks for efficient solid-phase extraction of sulfonamides in fish.

In this study, covalent organic frameworks (COFs) were grown in situ on magnetic nitrogen-doped graphene foam (MNGF), and the resulting composite of COFs-modified MNGF (MNC) was wrapped by molecularly imprinted polymers (MNC@MIPs) for specifically capturing SAs. A magnetic solid phase extraction (MSPE) method for SAs was established using MNC@MIPs with good magnetic responsiveness. The adsorption performance of MNC@MIPs was superior to that of non-molecularly imprinted polymers (MNC@NIPs), with shorter adsorption/desorption time and higher imprinting factors. A high-efficiency SAs analytical method was developed by fusing HPLC and MNC@MIPs-based MSPE. This approach provides excellent precision, a low detection limit, and wide linearity. By analyzing fish samples, the feasibility of the approach was confirmed, with SAs recoveries and relative standard deviations in spiked samples in the ranges of 77.2-112.7 % and 2.0-7.2 %, respectively. This study demonstrated the potential use of MNC@MIPs-based MSPE for efficient extraction and quantitation of trace hazards in food.

Atlantic salmon (Salmo salar) waste as a unique source of biofunctional protein hydrolysates: Emerging productions, promising applications, and challenges mitigation.

The Atlantic salmon is an extremely popular fish for its nutritional value and unique taste among several fish species. Researchers are focusing on the utilization of Atlantic salmon waste for generating protein hydrolysates rich in peptides and amino acids and investigating their health benefits. Several technological approaches, including enzymatic, chemical, and the recently developed subcritical water hydrolysis, are currently used for the production of Atlantic salmon waste protein hydrolysates. Hydrolyzing various wastes, e.g., heads, bones, skin, viscera, and trimmings, possessing antioxidant, blood pressure regulatory, antidiabetic, and anti-inflammatory properties, resulting in applications in human foods and nutraceuticals, animal farming, pharmaceuticals, cell culture, and cosmetics industries. Furthermore, future applications, constraints several challenges associated with industrial hydrolysate production, including sensory, safety, and economic constraints, which could be overcome by suggested techno processing measures. Further studies are recommended for developing large-scale, commercially viable production methods, focusing on eradicating sensory constraints and facilitating large-scale application.

Influence of increasing noise at the offshore wind farm area on fish vocalization phenology: A long-term marine acoustical monitoring off the foremost offshore wind farm in Taiwan.

The rapid increase of offshore projects at Taiwan Strait in recent decade has been debated for elevated noise levels. However, there are no studies on long-term assessment of noise levels and impact of noise on marine organisms. The passive acoustic monitoring was conducted at the foremost wind farm area in Taiwan to assess the sound levels and the impact of noise on fish vocalization behavior. Predominately, in the soundscape around the Taiwan Strait, two chorusing types (Type 1 and Type 2) from the Sciaenid family of fishes exist. Ambient sound levels significantly increased from 2014 to 2019, while the chorusing Types 1 and 2 were observed in a lower percentage of the recordings. Additionally, chorusing peak intensity and duration significantly reduced over the years for both choruses. This is the first field-based evidence to demonstrate the consequences of increasing anthropogenic noise having the potential to alter the vocalization behavior of the fish.

Hurricane-induced dramatic decline and natural recovery of a vulnerable sportfish population: Shoal bass (Micropterus cataractae) in the Chipola River, Florida.

Freshwater fishes are among the most biodiverse vertebrate groups and among the most threatened by anthropogenic activities. Many occur in small and geographically restricted populations that are increasingly subject to catastrophic events (hurricanes, wildfires, extreme floods and droughts), but it has rarely been possible to assess the impacts of such events. Here we document the decline and recovery of a regularly monitored, small shoal bass (Micropterus cataractae) population in the Chipola River, Florida following a catastrophic hurricane disturbance. The Chipola River population has the lowest level of interspecific hybridization (over 90 % non-introgressed shoal bass) within the species' range, a census population size of 2165 (95 % CI [1,383, 3,801]) in 2009 and a genetically effective population size Ne of 135 (95 % CI [70, 472]). In 2018, Hurricane Michael devastated the Chipola River and watershed. A survey conducted in 2019 indicated a severe decline (91 %) in relative population abundance and a very low Ne at 21 (95 % CI [16, 29]). However, the detection of young-of-year fish indicated that the depleted population had experienced successful reproduction. In fall 2021, the census population had recovered to 1039 fish (95 % CI [660, 1,814]) and Ne at 40 (95 % CI [31, 50]). While the population has shown considerable resilience in the face of hurricane disturbance, it remains vulnerable to future catastrophic events and may also suffer a long-term reduction in its adaptive potential due to a relatively low effective population size. To address these threats, continued monitoring is necessitated and targeted measures such as translocation of non-introgressed individuals from neighboring populations or establishment of a captive population of sufficient effective population size may be required to conserve the species in the long term.

Unravelling Stress Granules in the Deep Cold: Characterisation of TIA-1 Gene Sequence in Antarctic Fish Species.

Stress granules (SGs) are cytoplasmic foci lacking membranes, comprising non-translating messenger ribonucleoproteins, translational initiation factors, and additional proteins. Their formation is crucial for rapidly modulating gene expression in response to adverse environmental conditions, such as pollution and infections. Limited research has focused on investigating the molecular components of SGs in fish, with minimal exploration in Antarctic fish. This study characterises for the first time the transcript sequences of one key protein component of SGs, TIA-1 (T-cell intracellular antigen 1), in two Antarctic endemic fish species, i.e. Trematomus bernacchii and Chionodraco hamatus. The mRNA-binding protein TIA-1 acts as a post-transcriptional regulator of gene expression and its aggregation leads to the formation of SGs in response to cellular damage. The in vitro and bioinformatic analyses of the TIA-1 gene sequences of these two species highlighted interesting peculiarities, which include the transcription of alternatively spliced isoforms unique to the notothenioid lineage, potentially unlocking further insights into their unique adaptations to extreme environmental conditions. This is the first study to analyze tia-1 expression levels in different tissues of Antarctic fish species. Our key findings indicate that the TIA-1 gene is expressed at particularly high levels in the liver and spleen of C. hamatus, as well as in the heart and skeletal muscle of T. bernacchii. This suggests that those tissues play a significant role in the stress response mechanisms of the studied species. This study provides novel insights into the molecular adaptations of Antarctic fish, highlighting the potential importance of TIA-1 in their response to environmental stressors. The unique features of TIA-1 identified in these species may offer broader implications for understanding how Antarctic fish regulate gene transcriptions in their extreme environments.

Chemical, structural and antioxidant properties of melanoidins in fish sauce.

BACKGROUND: Melanoidins, as a functional component, exhibit antioxidant properties. However, the antioxidant mechanism of melanoidins in fish sauce remains poorly understood. The present study focused on the structural characteristics, microstructure and antioxidant activity of melanoidin substances in fish sauce. RESULTS: Ultrafiltration and color difference analysis were utilized to confirm the main molecular weight composition of melanoidins. The ultrafiltration component > 10 kDa in fish sauce exhibited the darkest and reddest color, along with the highest content of melanoidins (699.5 mg g-1). The spectral characteristics were consistent with typical melanoidins. Gas chromatography-tandem mass spectrometry and Fourier transform infrared spectroscopy analyses revealed that the melanoidins contained a significant number of oxygen-containing groups, amino compounds, carbohydrates, aromatic compounds and carbonyl compounds (CO), pyrrole (CN), among others. In total, 129 alkanes, 71 esters, nine olefins and two alkynes were identified. The melanoidins (> 10 kDa component) had the strongest antioxidant activity, including a reducing power of 0.8, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity of 67.7% and 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity of 92.4%. Additionally, the melanoidins (> 10 kDa component) had the highest total phenolic content at 48.2 µg gallic acid equivalents mL-1. CONCLUSION: Melanoidins are important factors affecting the antioxidant activity of fish sauce. There were differences in the structural properties of melanoidin fractions with different molecular weights. © 2024 Society of Chemical Industry.

Diving into the fish pathology of an important commercial fish species: the case of the European hake (Merluccius merluccius Linnaeus, 1758) in the northwest Mediterranean Sea.

The gross pathology and the histopathological alterations identified in juvenile specimens of the European hake (Merluccius merluccius), one of the most important target species of commercial fisheries in the northwest Mediterranean, are described. A qualitative and semi-quantitative histological approach was performed in specimens from 2007 and 2019. Prevalence and mean intensity of parasites and histopathological changes were calculated in both years. No macroscopic alterations were found in any organ but several parasites (e.g., copepods, nematodes, digeneans) were detected. Microscopically, alterations identified in gills included foci of inflammation and hyperplasia (present in 77.38% of hakes from both years), extensive hyperplasia (33.33%), and inflammation (16.65%) that were potentially related to the mechanical effects of monogeneans and copepods, cysts of unknown etiology (62.69%), and lamellar inflammation associated to Aporocotyle spinosicanalis eggs (8.33%). Granulomas and inflammatory focus were detected in the liver, spleen, and stomach, apparently associated with the presence of nematodes. Coelozoic myxosporean parasites were detected within the renal tubules (66.66%). Most of the pathologies detected were similar to those described in other gadoid species (i.e., Gadus morhua) and were usually related to the presence of ecto and endoparasites. The potential impact of parasites on the health of this fish species is discussed for improving the knowledge and management of these valuable fishing stocks.

Expression and regulation of the CXCL9-11 chemokines and CXCR3 receptor in Atlantic salmon (Salmo salar).

Chemokines are cytokines that mediate leukocyte traffic between the lymphoid organs, the bloodstream, and the site of tissue damage, which is essential for an efficient immune response. In particular, the gamma interferon (IFN- γ) inducible chemokines CXCL9, CXCL10, and CXCL11, and their receptor CXCR3, are involved in T cell and macrophage recruitment to the site of infection. The nature and function of these chemokines and their receptor are well-known in mammals, but further research is needed to achieve a similar level of understanding in fish immunity. Thus, in this study, we seek to identify the genes encoding the components of the Atlantic salmon (Salmo salar) CXCL9, CXCL10, CXCL11/CXCR3 axis (CXCL9-11/CXCR3), predict the protein structure from the amino acid sequence, and explore the regulation of gene expression as well as the response of these chemokines and their receptor to viral infections. The cxcl9, cxcl10, cxcl11, and cxcr3 gene sequences were retrieved from the databases, and the phylogenetic analysis was conducted to determine the evolutionary relationships. The study revealed an interesting pattern of clustering and conservation among fish and mammalian species. The salmon chemokine sequences clustered with orthologs from other fish species, while the mammalian sequences formed separate clades. This indicates a divergent evolution of chemokines between mammals and fish, possibly due to different evolutionary pressures. While the structural analysis of the chemokines and the CXCR3 receptor showed the conservation of critical motifs and domains, suggesting preserved functions and stability throughout evolution. Regarding the regulation of gene expression, some components of the CXCL9-11/CXCR3 axis are induced by recombinant gamma interferon (rIFN-γ) and by Infectious pancreatic necrosis virus (IPNV) infection in Atlantic salmon cells. Further studies are needed to explore the role of Atlantic salmon CXCL9-11 chemokines in regulating immune cell migration and endothelial activation, as seen in mammals. To the best of our knowledge, there have been no functional studies of chemokines to understand these effects in Atlantic salmon.

Laying the foundations for selective-fish guidance using electricity: multi-species response to pulsed direct currents.

To develop effective technology that employs electric fields to simultaneously guide valued freshwater fish whilst limiting the range expansion of undesirable invasive species, there is a need to quantify the electrosensitivity of multiple families. This experimental study quantified the electrosensitivity of two carp species that, in UK, are invasive (grass carp, Ctenopharyngodon idella, and common carp, Cyprinus carpio) and compared the values with those previously obtained for adult European eel (Anguilla anguilla), a species of conservation concern in Europe. Electric field strengths (V/cm) required to elicit physiological responses (twitch, loss of orientation and tetany) were identified across four pulsed direct current (PDC) electric waveforms (single pulse-2 Hz, double pulse-2 Hz, single pulse-3 Hz and double pulse-3 Hz). Grass carp were sensitive to differences in waveform with tetany exhibited at lower field strengths in the single pulse-2 Hz treatment. Both cyprinid species responded similarly and were less sensitive to PDC than adult European eel, although loss of orientation occurred at lower field strengths for grass than common carp in the single pulse-3 Hz waveform treatment. This variation in electrosensitivity, likely due to differences in body length, indicates potential for electric fields to selectively guide fish in areas where invasive and native species occur in sympatry.

Alternative protein sources in aquafeed: Current scenario and future perspectives.

Fish meal represents the main protein source for most commercially farmed aquatic species, as it is characterized by high nutritional value and lack of anti-nutritional factors. However, its availability and the market price have been recognized as serious problems at least for over a decade, making it necessary to search for non-conventional protein sources, as an alternative to fish meals. This review aims to comprehensively examine and critically revise the use of fish meal and all alternative protein sources explored to date on the health, welfare, and growth performance of the major aquatic species commercially interesting from a global scenario. The investigation revealed that the inclusion levels of the different protein sources, plant- and animal-derived, ranged from 10 to 80 % and from 2 to 100 % respectively, in partial or complete replacement of fish meal, and generated positive effects on health, welfare, growth performance, and fillet quality. However, the results showed that above a certain level of inclusion, each protein source can negatively affect fish growth performance, metabolic activities, and other biological parameters. Moreover, it is likely that by mixing different protein sources, the combination of each ingredient causes a synergistic effect on the nutritional properties. Therefore, the future of aquatic feed formulation is expected to be based on the blend of different protein sources. Overall, the analysis highlighted the need for additional research in the field of replacing fish meals with new protein sources, given that many knowledge gaps are still to be filled on aquatic species, which deserve to be investigated.

Characterization of Lactic Acid Bacteria Isolated From Rotting Oranges and Use of Agropastoral Processing By-products as Carbon and Nitrogen Sources Alternative for Lactic Acid Production.

This study investigated the ability of lactic acid bacteria (LAB) isolated from oranges to use fish by-products (FB) and chicken by-products (CB) as nitrogen sources alternative to yeast extract for lactic acid (LA) production in a papaya by-product medium as a carbon source. Once the fermentation agents had been isolated, they were subjected to biochemical and molecular characterization. Inexpensive nitrogen sources, precisely CB and FB, were prepared, freeze-dried, and yield evaluated. Also, before to the fermentation experiments, the Total Kjehdahl Nitrogen (TKN) of these by-products and that of the yeast extract were determined. Then, three production media differing in terms of nitrogen source were formulated from these nitrogen sources. From the 22 LAB isolated from orange, two isolates of interest (NGO25 and NGO23) were obtained; all belonging to the Lactiplantibacillus plantarum species based on 16S rRNA gene sequencing. Furthermore, the production yield powder obtained after lyophilization of 1 L of CB and FB surpernatant were, respectively, 16.6 g and 12.933 g. The TKN of different nitrogen sources powder were 71.4 ± 0.000% DM (FB), 86.145 ± 0.001% DM (CB), and 87.5 ± 0.99% DM (yeast extract). The best kinetic parameters of LA production (LA (g/L): 31.945 ± 0.078; volumetric productivity (g/L.h): 1.331 ± 0.003; LA yield (mg/g) 63.89 ± 0.156; biomass (g/L) 7.925 ± 0.035; cell growth rate (g/L.h): 0.330 ± 0.001) were recorded by Lactiplantibacillus plantarum NGO25 after 24 h of fermentation. The latter data were obtained in the production medium containing CB as nitrogen sources. In addition, this production medium cost only $0.152 to formulate, compared to yeast extract which required $1.692 to formulate. Thus, freeze-dried CB can be used as an alternative to yeast extract in large-scale production of LA.

Effectiveness of fish oil in controlling inflammation in adult patients undergoing hemodialysis: A systematic review and meta-analysis.

Objective: Hemodialysis patients with chronic kidney disease often exhibit inflammation characterized by elevated levels of C-reactive protein, Interleukin 6 and tumor necrosis factor-alpha, and they are shown to be associated with cardiovascular impairment and enhanced renal failure. This study aims to assess the impact of fish oil intake on inflammation indicators in adult hemodialysis patients. Methods: From the inception to December 2023, the datasets Cochrane Central, Google Scholar, Science Direct, Embase, and Pubmed were examined. Two authors independently searched, selected, and screened the literature. The pooled results are represented by weighted mean difference (WMD) with 95% confidence intervals. To investigate the causes of heterogeneity, subgroup analysis was done. Sensitivity analysis was then used to evaluate the validity of the combined findings. Results: Thirteen randomized control trials studies were included. The pooled results showed that fish oil supplementation caused a significant reduction of the C-reactive protein level (WMD, -2.92 mg/L; 95% Confidence interval, -5.23, to -0.61; p = 0.01; I 2 = 99%), especially in patients with baseline C-reactive protein ⩾5 mg/L (WMD, -4.39 mg/L; 95% Confidence interval, -5.93 to 2.85; p < 0.00001; I 2 = 33%). Subgroup analyses showed that C-reactive protein baseline level (C-reactive protein <5 mg/L) was the main source of heterogeneity. Fish oil intake may not reduce the level of Interleukin 6 (WMD, -2.26; 95% Confidence interval: -19.61 to 15.09; p = 0.80; I 2 = 93%), nor will it reduce the level of tumor necrosis factor-alpha (random model: WMD, -2.51; 95% Confidence interval: 6.08 to 1.06; p = 0.17; I 2 = 98%). Conclusion: Hemodialysis patients, especially those with C-reactive protein > 5 mg/L, responded to fish oil supplementation to reduce their C-reactive protein level; however, Interleukin 6 and tumor necrosis factor-alpha levels did not appear to be affected.

First record of the rare tubeshoulder Holtbyrnia anomala Krefft 1980 (Teleostei, Platytroctidae) in the southwest Atlantic.

Holtbyrnia anomala is a bathypelagic platytroctid widely distributed in the Atlantic Ocean. In this contribution, we report, for the first time, the occurrence of this species in the tropical southwest Atlantic. A single specimen was collected in 2000 on the continental slope off Rio de Janeiro, Brazil, at an average depth of 1158 m. This report also represents the first record of Holtbyrnia anomala in the Brazilian Economic Exclusive Zone.

Evaluation of stress in farmed Atlantic salmon (Salmo salar) using different biological matrices.

Atlantic salmon were subjected to an acute crowding scenario, and their subsequent stress responses were observed under three distinct swimming speed/water flow (WF) conditions: 0.5, 1, and 1.5 body lengths per second (BL/s). Feces, dermal mucus, and plasma were collected for analysis at 1, 6, and 24 h (h) post-stress. Additionally, the head kidney and two regions of the brain (pituitary and POA) were collected for transcript expression analysis. Fish swimming at 0.5 BL/s exhibited higher pre-stress (baseline) cortisol levels. Across all groups and matrices, the highest cortisol/cortisol metabolites (CM) levels were observed at the 1 h post-stress sampling point. At 6 h (second sampling time point), a clear decline toward baseline levels were observe in all groups. Significant increases in mean plasma glucose levels were observed at 1 h post-stress for all groups. The mean plasma lactate levels varied based on WF treatments, with a significant increase observed at 1 h only for the 1.5 BL/s group. Additionally, significant decreases in mean plasma lactate were noted at 6 and 24 h post-stress for some groups. The mRNA abundances of the tested genes (star, cyp17a1, hsd11ß2, srd5a1) increased following the stress events. These changes were not uniform across all groups and were tissue dependent. In summary, the results indicate that mucus and feces can be used as potentially less invasive matrices than blood for evaluating stress and, consequently, the welfare of Atlantic salmon in captivity.

Faster ciguatoxin extraction methods for toxicity screening.

Ciguatera poisoning (CP) is a severe global public health problem caused by the consumption of seafood products contaminated with ciguatoxins (CTXs). The growing demand for seafood products requires high-throughput testing for CTX-susceptible seafood, however complex extraction and slow cleanup methods inhibit this goal. Herein, several methods for extracting CTXs from fish tissue were established and compared; these methods are sensitive, specific, and valid while achieving higher sample extraction throughput than currently established protocols. The trial fish material was generated from multiple species, with different physical conditions (wet and freeze-dried tissue), and naturally contaminated with various CTXs (i.e., CTX-1B, CTX-3C, and C-CTX-1), thus ensuring these methods are robust and broadly applicable. The extraction methods used were based on mechanical maceration with acetone or methanol or enzymatic digestion followed by acetone and ethyl acetate extraction. Crude extracts were investigated for CTX-like toxicity using an in vitro mouse neuroblastoma (N2a) cell-based assay (CBA). Among the three methods, there was no significant difference in toxin estimates (p = 0.219, two-way ANOVA), indicating their interchangeability. For speed (> 16 samples/day), accuracy (100%), and CTX analog retention confirmation by liquid chromatography-tandem mass spectrometry (LC‒MS/MS), the preferred extraction methods were both methanol and enzyme-based. All extraction methods post hoc confirmation of CTX analogs successfully met international seafood market-based CTX contaminant guidance. These methods can drastically increase global CTX screening capabilities and subsequently relieve sample processing bottlenecks, inhibiting environmental and human health-based CTX analysis.

The costs of implementing anaemia reduction interventions among women fish processors in Ghana.

BACKGROUND: Anaemia among women of reproductive age (WRA) remains persistently high in Ghana, affecting 41% in 2022. Women in low-income communities in Ghana engaged in fish processing activities are at increased risk of anaemia due to inadequate diets, exposure to infectious pathogens, and pollutants. The Invisible Fishers (IFs) project was implemented among women fish processors in their reproductive age in Central and Volta regions of Ghana to mitigate anaemia. Despite the efficacy, feasibility and scalability of the intervention, the cost of implementing the intervention is unknown. The objective of this study was to estimate the costs of implementing the IFs project in Ghana. METHODS: We used micro-costing approach to analyse the costs of implementing the IFs project. Data were collected as part of a pilot randomized control trial with three interventions: Behaviour Change Communication (BCC), Strengthening Market Engagement of fish processors plus Behaviour Change Communication (SME + BCC), and Fish Smoking Technology and Practices plus Behaviour Change Communication (FST + BCC). The interventions were delivered to 60 women fish processors in the Central region and 60 in Volta region. The cost of the intervention was estimated from the societal perspective. Economic costs were categorized as direct costs (i.e. personnel, transportation, meetings, training, and monitoring) and indirect cost (i.e. value of productive time lost due to women and community volunteers' participation in the activities of the IFs project). RESULTS: The FST + BCC had the highest average cost per beneficiary (US$11898.62), followed by the SME + BCC (US8962.93). The least expensive was the BCC (US$4651.93) over the intervention period of 18 months. Recurrent costs constituted the largest component of economic costs (98%). Key drivers of direct costs were personnel (58%), administrative expenses (14%), and transportation (7%). CONCLUSION: There is a high cost for implementing interventions included in the IFs project. Planning and scaling -up of the interventions across larger populations could bring about economies of scale to reduce the average cost of the interventions.

New insights into the phylogeny of Carasobarbus Karaman, 1971 (Actinopterygii, Cyprinidae) with the description of three new species.

Fishes from the genus Carasobarbus, widely distributed throughout the river systems of North Africa and West Asia, are commonly referred to as Himris. In the Persian Gulf basin, they are widespread and are also found in fast-flowing rivers or the deeper regions of lakes. In this region, representation of these fishes in scientific collections is scarce, and except for C. luteus, the other species are very poorly documented and frequently misidentified due to their similarities. In this study we analysed the relationships among Carasobarbus species using mitochondrial genes (Cyt b, COI) and present morphological characters based on examinations. Our results revealed three new species which we describe here. Carasobarbus doadrioi, new species, is distinguished by 40-44 scales on the lateral line and a prominent black blotch on end of caudal peduncle in specimens < 85 mm SL. Carasobarbus hajhosseini, new species is distinguished by 32-34 scales on the lateral line and long head length (20-24% SL). Carasobarbus saadatii, new species, is distinguished by 38-40 scales on the lateral line and short head length (19-20% HL). In the Persian Gulf basin, Carasobarbus species exhibit uncorrected genetic distances of 1.6 to 5.5% in the COI barcode region and 2.6% to 9.9% in the Cyt b gene. This study highlights the importance of investigating the unexplored diversity that exists within poorly sampled and understudied freshwater fish group. Such investigations are essential for developing a comprehensive understanding of the true extent of biodiversity, which is critical for informing effective conservation and protection strategies.