The Replacement of Fish Meal with Poultry By-Product Meal and Insect Exuviae: Effects on Growth Performance, Gut Health and Microbiota of the European Seabass, Dicentrarchus labrax.

This study addressed the urgent need for sustainable protein sources in aquaculture due to the depletion of marine resources and rising costs. Animal protein sources, particularly poultry by-product meal (PBM) and insect exuviae meal, were investigated as viable alternatives to fishmeal (FM). The research study confirmed the successful replacement of FM with a combination of PBM and insect exuviae meal (up to 50%) in the diet of European seabass without compromising growth, feed conversion, gut health, and liver fat content. In particular, growth was robust with both PBM formulations, with the 25% PBM diet showing better results. Histological examinations showed good gut and liver health, contradicting the concerns of previous studies. This paper emphasizes the importance of holistic analyzes that go beyond growth parameters and include histomorphological investigations. The results show that PBM in combination with insect/exuviae meal is well tolerated by seabass, which is consistent with reports in the literature of it mitigating negative effects on gut health. A detailed analysis of the microbiota revealed a decrease in the Firmicutes/Proteobacteria ratio due to an increase in potentially pathogenic bacteria. However, the formulation containing insect exuviae partially counteracted this effect by preserving the beneficial Lactobacillus and promoting the synthesis of short-chain fatty acids (SCFAs), particularly butyrate. Chitin-rich components from insect exuviae were associated with improved gut health, which was supported by the increased production of SCFAs, which are known for their anti-inflammatory properties. This paper concludes that a combination of PBM and insect/exuviae meal can replace up to 50% of FM in the diet of seabass, supporting sustainable aquaculture practices. Despite some changes in the microbiota, the negative effects are mitigated by the addition of insect exuviae, highlighting their potential as a prebiotic to increase fish productivity and contribute to a circular economy in aquaculture.

The impact of diets containing Hermetia illucens meal on the growth, intestinal health, and microbiota of gilthead seabream (Sparus aurata).

The present study investigated the effect of replacing fishmeal (FM) with insect meal of Hermetia illucens (HI) in the diet of Sparus aurata farmed inshore on growth, gut health, and microbiota composition. Two isolipidic (18% as fed) and isoproteic (42% as fed) diets were tested at the farm scale: a control diet without HI meal and an experimental diet with 11% HI meal replacing FM. At the end of the 25-week feeding trial, final body weight, specific growth rate, feed conversion rate, and hepatosomatic index were not affected by the diet. Gross morphology of the gastrointestinal tract and the liver was unchanged and showed no obvious signs of inflammation. High-throughput sequencing of 16S rRNA gene amplicons (MiSeq platform, Illumina) used to characterize the gut microbial community profile showed that Proteobacteria, Fusobacteria, and Firmicutes were the dominant phyla of the gut microbiota of gilthead seabream, regardless of diet. Dietary inclusion of HI meal altered the gut microbiota by significantly decreasing the abundance of Cetobacterium and increasing the relative abundance of the Oceanobacillus and Paenibacillus genera. Our results clearly indicate that the inclusion of HI meal as an alternative animal protein source positively affects the gut microbiota of seabream by increasing the abundance of beneficial genera, thereby improving gut health and maintaining growth performance of S. aurata from coastal farms.

Polychaete (Alitta virens) meal inclusion as a dietary strategy for modulating gut microbiota of European seabass (Dicentrarchus labrax).

Recent research has revealed the significant impact of novel feed ingredients on fish gut microbiota, affecting both the immune status and digestive performance. As a result, analyzing the microbiota modulatory capabilities may be a useful method for assessing the potential functionality of novel ingredients. Therefore, this study aimed to evaluate the effects of dietary polychaete meal (PM) from Alitta virens on the autochthonous and allochthonous gut microbiota of European seabass (Dicentrarchus labrax). Two diets were compared: a control diet with 25% fishmeal (FM) and a diet replacing 40% of fishmeal with PM, in a 13-week feeding trial with juvenile fish (initial weight of 14.5 ± 1.0 g). The feed, digesta, and mucosa-associated microbial communities in fish intestines were analyzed using high-throughput sequencing of the 16S rRNA gene on the Illumina MiSeq platform. The results of feed microbiota analyses showed that the PM10 feed exhibited a higher microbial diversity than the FM diet. However, these feed-associated microbiota differences were not mirrored in the composition of digesta and mucosal communities. Regardless of the diet, the digesta samples consistently exhibited higher species richness and diversity than the mucosa samples. Overall, digesta samples were characterized by a higher abundance of Firmicutes in PM-fed fish. In contrast, at the gut mucosa level, the relative abundances of Mycobacterium, Taeseokella and Clostridium genera were lower in the group fed the PM10 diet. Significant differences in metabolic pathways were also observed between the FM and PM10 groups in both mucosa and digesta samples. In particular, the mucosal pathways of caffeine metabolism, phenylalanine metabolism, and sulfur relay system were significantly altered by PM inclusion. The same trend was observed in the digesta valine, leucine, and isoleucine degradation and secretion pathways. These findings highlight the potential of PM as an alternative functional ingredient in aquafeeds with microbiota modulatory properties that should be further explored in the future.

Aquaculture ecosystem microbiome at the water-fish interface: the case-study of rainbow trout fed with Tenebrio molitor novel diets.

BACKGROUND: Sustainable aquaculture relies on multiple factors, including water quality, fish diets, and farmed fish. Replacing fishmeal (FM) with alternative protein sources is key for improving sustainability in aquaculture and promoting fish health. Indeed, great research efforts have been made to evaluate novel feed formulations, focusing especially on the effects on the fish gut microbiome. Few studies have explored host-environment interactions. In the present study, we evaluated the influence of novel insect-based (Tenebrio molitor) fish diets on the microbiome at the water-fish interface in an engineered rainbow trout (Oncorhynchus mykiss) farming ecosystem. Using 16S rRNA gene metabarcoding, we comprehensively analyzed the microbiomes of water, tank biofilm, fish intestinal mucus, fish cutis, and feed samples. RESULTS: Core microbiome analysis revealed the presence of a highly reduced core shared by all sample sources, constituted by Aeromonas spp., in both the control and novel feed test groups. Network analysis showed that samples were clustered based on the sample source, with no significant differences related to the feed formulation tested. Thus, the different diets did not seem to affect the environment (water and tank biofilm) and fish (cutis and intestinal mucus) microbiomes. To disentangle the contribution of feed at a finer scale, we performed a differential abundance analysis and observed differential enrichment/impoverishment in specific taxa, comparing the samples belonging to the control diet group and the insect-based diet group. CONCLUSIONS: Omic exploration of the water-fish interface exposes patterns that are otherwise undetected. These data demonstrate a link between the environment and fish and show that subtle but significant differences are caused by feed composition. Thus, the research presented here is a step towards positively influencing the aquaculture environment and its microbiome.

Sustainable Fish Feeds with Insects and Probiotics Positively Affect Freshwater and Marine Fish Gut Microbiota.

Aquaculture is the fastest-growing agricultural industry in the world. Fishmeal is an essential component of commercial fish diets, but its long-term sustainability is a concern. Therefore, it is important to find alternatives to fishmeal that have a similar nutritional value and, at the same time, are affordable and readily available. The search for high-quality alternatives to fishmeal and fish oil has interested researchers worldwide. Over the past 20 years, different insect meals have been studied as a potential alternate source of fishmeal in aquafeeds. On the other hand, probiotics-live microbial strains-are being used as dietary supplements and showing beneficial effects on fish growth and health status. Fish gut microbiota plays a significant role in nutrition metabolism, which affects a number of other physiological functions, including fish growth and development, immune regulation, and pathogen resistance. One of the key reasons for studying fish gut microbiota is the possibility to modify microbial communities that inhabit the intestine to benefit host growth and health. The development of DNA sequencing technologies and advanced bioinformatics tools has made metagenomic analysis a feasible method for researching gut microbes. In this review, we analyze and summarize the current knowledge provided by studies of our research group on using insect meal and probiotic supplements in aquafeed formulations and their effects on different fish gut microbiota. We also highlight future research directions to make insect meals a key source of proteins for sustainable aquaculture and explore the challenges associated with the use of probiotics. Insect meals and probiotics will undoubtedly have a positive effect on the long-term sustainability and profitability of aquaculture.

Effect of 2400 MHz mobile phone radiation exposure on the behavior and hippocampus morphology in Swiss mouse model.

Electromagnetic field exposure to the nervous system can cause neurological changes. The effects of extremely low-frequency electromagnetic fields, such as second-generation and third-generation radiation, have been studied in most studies. The current study aimed to explore fourth-generation cellular phone radiation on hippocampal morphology and behavior in mice. Swiss albino male mice (n = 30) were randomly categorized into 3 groups; control, 40 min, and 60 min exposure to 2400 MHz radiofrequency electromagnetic radiation (RF-EMR) daily for 60 days. The control mice were housed in the same environments but were not exposed to anything. Anxiety-like behaviors were tested using the elevated plus-maze. For histological and stereological examination, the brain was dissected from the cranial cavity. On Cresyl violet stained brain slices, the number of pyramidal neurons in the cornu ammonis of the hippocampus were counted. In exposed mice compared to control mice, a significant increase in anxiety-like behavior has been observed. Histological observations have shown many black and dark blue cytoplasmic cells with shrunken morphology degenerative alterations in the neuronal hippocampus in the radiation exposed mice. In the RF-EMR mouse hippocampus, stereological analyses revealed a significant decrease in pyramidal and granule neurons compared to controls. Our findings suggest that 2400-MHz RF-EMR cell phone radiation affects the structural integrity of the hippocampus, which would lead to behavioral changes such as anxiety. However, it alerts us to the possible long-term detrimental effects of exposure to RF-EMR.

Immunophenotypic analysis of the distribution of hepatic macrophages, lymphocytes and hepatic stellate cells in the adult rat liver.

The liver consists of parenchymal hepatocytes and non-parenchymal cells. Non-parenchymal cells, Kupffer cells, hepatic stellate cells and cholangiocytes have crucial roles in liver homeostasis and liver pathology. To establish baseline data, this study investigated immunohistochemically the distribution of non-parenchymal cells in perivenular areas (PV), periportal areas (PP) and Glisson's sheath (GS) of adult rat liver. Liver tissues were collected from the left lateral lobe of rats. CD163-positive macrophages were seen along the sinusoid of PV and PP areas, indicating Kupffer cells. Double immunofluorescence showed, Kupffer cells partly co-expressed CD68 and MHC class II antigens in the liver. The numbers of Kupffer cells were significantly high in PP areas as compared with PV or GS areas. CD68-positive exudative macrophages were highly localized in PP and GS areas and a comparatively low PV area. MHC class II-positive dendritic cells (activated macrophages) were localized mainly in GS. Granzyme B-positive NK cells were mainly localized in the Glisson's sheath. CD3-positive T cells and CD20-positive B cells were distributed along the sinusoids of the PP and PV areas of hepatic lobules. Vimentin and glial fibrillary acidic protein (GFAP)-positive hepatic stellate cells were localized along sinusoids in the hepatic lobules of the liver. Cholangiocytes reacting to cytokeratin 19 were seen on interlobular bile ducts in Glisson's sheath of the liver. This study shows that heterogeneous macrophage populations, liver-resident lymphocytes and hepatic stellate cells localized in PP and PV areas or GS areas of the liver with cells specific patterns.

Effect of formaldehyde and urea contaminated feed exposure into the liver of young and adult pigeons (Columba livia).

BACKGROUND AND AIM: Nowadays, toxic chemical contaminants in food are a major food safety problem in Bangladesh. Among toxic food contaminants, formalin is used to preserve fruit, vegetables, and fish, where urea is used for the whitening of rice and puffed rice. The purpose of this study was to determine the biochemical and histopathological effects on the liver of young and adult pigeons after exposure to formalin and urea contaminated feed. MATERIALS AND METHODS: A total of 15 young and 15 adult pigeons were divided into control group, formaldehyde exposed group (2.5 mL formalin/kg feed), and urea exposed (1 g/kg feed) group. Each group consisted of five pigeons. After the experimentation procedures, the blood samples were collected for biochemical study, and the liver tissue was collected for histomorphological study. The statistical analysis was performed using the Student's t-test, and p<0.05 was considered as statistically significant. RESULTS: The aspartate transaminase serum hepatic enzyme was significantly increased in both formalin and urea exposed young and adult pigeons than the control pigeons. In control pigeons, parenchymal hepatocytes and non-parenchymal cells are regularly arranged. However, histological observation of the liver of formalin and urea exposed young, and adult pigeons showed coagulation necrosis with infiltration of many inflammatory cells around the central and portal veins. The necrotic areas are more extensive with massive infiltration of inflammatory cells in the liver of formalin-treated pigeons than the urea treated pigeons. CONCLUSION: The present study results show that low concentrations of formalin and urea in feed induced liver lesions in pigeons in different extents and indicate that exposure to toxic chemicals may affect homeostasis of the liver and cause liver injury or act as a co-factor for liver disease.

Hematobiochemical and histopathological alterations of kidney and testis due to exposure of 4G cell phone radiation in mice.

The radiofrequency electromagnetic radiation emitted by smart phones on biological systems has wide media coverage and public concern in recent years. The aim of this study was to explore the effects of fourth-generation cell phone radiation exposure on hematological (Total leukocyte count, Total erythrocyte count, and hemoglobin %), biochemical (Serum creatinine) parameters, and histopathological changes in the kidney and testis of Swiss albino mice. A total of 30 male Swiss albino mice weighing 45-65 g was randomly divided into three groups (n = 10). The first group A was the control group, the second group B, was exposed to 40 minutes of mobile phone radiation daily, the third group C was exposed to 60 minutes of radiation daily from two 2400 Megahertz fourth-generation connected mobile phones for 60 days, respectively. The electromagnetic radiation frequency radiometer measured the frequency of electromagnetic radiation emitted from cell phones. The specific absorption rate was calculated as 0.087 W/kg. The control group was kept under similar conditions, but the electromagnetic field was not given for the same period. All the mice were sacrificed at the end of the experiment. The blood samples were collected for hematobiochemical study, and then kidney and testis tissues were collected for histopathological study. Results of the study showed that the body weight and total erythrocyte count values were significantly (p < 0.05) decreased while total leukocyte count, hemoglobin %, and serum creatinine values were significantly (p < 0.05) increased in both the radiation exposure groups relative to the control group. Histopathological observation showed the kidney of 60 minutes exposed mice interstitial inflammation that causes marked mononuclear cellular infiltration compared to the 40 minutes and control mice. Compared to control mice, histopathological examinations of testicular tissue from the exposed mice, showed irregular in shapes and non-uniform sizes and fewer spermatogenic cells layer that leads to the larger lumen in the seminiferous tubules. It is concluded that fourth-generation cell phone radiation exposure may affect blood hemostasis and inflammation of mice's kidney and testis tissue. Based on these studies, it is important to increase public consciousness of potential adverse effects of mobile phone radiofrequency electromagnetic radiation exposure.

Biochemical and histopathological effects of mobile phone radiation on the liver of Swiss albino mice

The electromagnetic radiation from the mobile phone is a subject of recent study because of the enormous increase in mobile phone use through-out the world. The objective of this experiment was therefore to investigate the biochemical and histopathological effects of mobile phone radiation on male Swiss albino mice’s liver. Male mice were categorized into three groups in this research: control group (A), exposed group for 40 minutes (B) and exposed group for 60 minutes (C). Experimental groups were exposed to radiation per day for 60 days from 4G connected mobile phones. The control group received no radiation. At the end of the radiation exposure, biochemical (alanine transaminase (ALT) and aspartate transaminase (AST)) and histological tests were performed. The results indicated that there was significant (P < 0.05) increase in mean values of ALT and AST in both radiation-exposed groups of mice if com-pared to the control group. Histopathologically marked infiltration of mononuclear cellular aggregates were present surrounding the bile duct and hepatic artery in the liver of 60-minute-exposure group, whereas in 40-minute-exposure group con-gestion was observed in the portal vein and the central vein of the liver. The findings revealed and evidenced that mobile phone radiation has harmful effects on enzyme activity and liver tis

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Ascertainment of pharmacological activities of Allamanda neriifolia Hook and Aegialitis rotundifolia Roxb used in Bangladesh: An in vitro study.

The present study was cherished to investigate in vitro thrombolytic, membrane stabilizing and antibacterial activities of Allamanda neriifolia and Aegialitis rotundifolia. Different types methanolic extracts of these two medicinal plants were tested for determining membrane stabilizing activity at a hypotonic solution and heat induce condition by comparing with reference standard acetyl salicylic acid (0.10 mg/mL), where thrombolytic activity assessment was done by employing Streptokinase as standard drug. Finally, antibacterial activity was performed against Staphylococcus aureus as a Gram-positive (+ve) and Salmonella typhi, Escherichia coli and Pseudomonas aeruginosa as Gram negative (-ve) bacteria by using disc diffusion method. In case of membrane stabilizing studies, crude methanolic extracts of A. neriifolia at 10 mg/ml concentration, more importantly, showed 45.80% & 23.52% whereas 10 mg/ml concentration of A. rotundifolia more significantly (p < 0.01) produced 38.40% and 27.04% inhibition of hemolysis for both experimental conditions. Dose-dependently increased activity was found in the thrombolytic study where 10 mg/ml concentration of both A. neriifolia and A. rotundifolia more significantly (p < 0.01) showed 41.91% and 32.76% lysis of clot respectively by in vitro clot lysis assay method. Crude methanolic extracts of A. rotundifolia did not show any suitable antibacterial property against the test bacteria. However, the gram positive (+ve) bacteria also seemed resistant against A. neriifolia extract but this crude methanolic extracts was found to generate moderate antibacterial action against gram-negative (-ve) bacteria. The obtained results confirmed the presence of thrombolytic, membrane stabilizing activity for both plant extract along with moderate antibacterial activity for A. neriifolia.

The "double endoloop" technique--a simple alternative technique for laparoscopic appendectomy.

Diagnostic laparoscopy is now being frequently used in the emergency management of patients with acute lower abdominal pain, particularly where appendicitis is suspected. It is particularly useful in women where other responsible gynecologic causes can be diagnosed and treated thereby decreasing the rate of negative open appendectomy. Recent advances in laparoscopic techniques have resulted in increasing numbers of patients proceeding to laparoscopic appendectomy. Here, we describe a modification of laparoscopic appendectomy using a "double endoloop technique." We have used this method in 53 patients during last 18 months in selected cases. The procedure is simple, quick, effective, and cheap.