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Generation of double knockout cattle via CRISPR-Cas9 ribonucleoprotein(RNP)electroporation

Journal of Animal Science and Biotechnology 2024年第1期15卷 456-462页

作者： Gyeong-Min Gim Kyeong-Hyeon Eom Dong-Hyeok Kwon Dae-Jin Jung Dae-Hyun Kim Jun-Koo Yi Jae-Jung Ha Ji-Hyun Lee Seong-Beom Lee Woo-Jae Son Soo-Young Yum Won-Wu Lee Goo Jang Laboratory of Theriogenology and Biotechnology Department of Veterinary Clinical ScienceCollege of Veterinary Medicine and the Research Institute of Veterinary ScienceSeoul National UniversitySeoulRepublic of Korea LARTBio Co. Ltd.SeoulRepublic of Korea Gyeongsangbukdo Livestock research Institute YeongjuRepublic of Korea Comparative Medicine Disease research center Seoul National UniversitySeoulRepublic of Korea

Background Genome editing has been considered as powerful tool in agricultural ***,genome editing progress in cattle has not been fast as in other mammal species,for some disadvantages including long gestational periods,single pregnancy,and high raising ***,technically demanding methods such as microinjection and somatic cell nuclear transfer(SCNT)are needed for gene editing in *** this point of view,electroporation in embryos has been risen as an *** First,editing efficiency of our electroporation methods were tested for *** of mutation on embryo was confirmed by T7E1 *** first combination,mutation rates for MSTN and PRNP were 57.6%±13.7%and 54.6%±13.5%,*** case of MSTN/BLG,mutation rates were 83.9%±23.6%for MSTN,84.5%±18.0%for ***,the double-KO embryos were transferred to surrogates and mutation rate was identified in resultant calves by targeted deep *** recipients were transferred for MSTN/PRNP,4 calves were delivered,and one calf underwent an induction for double *** surrogates were given double-KO embryos for MSTN/BLG,and four of the six calves that were born had mutations in both *** These data demonstrated that production of genome edited cattle via electroporation of RNP could be effectively ***,MSTN and PRNP from beef cattle and MSTN and BLG from dairy cattle have been born and they will be valuable resources for future precision breeding.

关键词： Beta-lactoglobulin Cattle CRISPR-Cas9 Electroporation Knockout MSTN PRNP

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Understanding the spectrum of non-motor symptoms in multiple sclerosis:insights from animal models

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Neural Regeneration research 2024年第1期19卷 84-91页

作者： Poornima D.E.Weerasinghe-Mudiyanselage Joong-Sun Kim Taekyun Shin Changjong Moon department of veterinary Anatomy and Animal Behavior College of Veterinary Medicine and BK21 FOUR programChonnam National UniversityGwangjuRepublic of Korea department of veterinary Anatomy College of Veterinary Medicine and Veterinary Medical Research InstituteJeju National UniversityJejuRepublic of Korea

Multiple sclerosis is a chronic autoimmune disease of the central nervous system and is generally considered to be a non-traumatic,physically debilitating neurological *** addition to experiencing motor disability,patients with multiple sclerosis also experience a variety of nonmotor symptoms,including cognitive deficits,anxiety,depression,sensory impairments,and ***,the pathogenesis and treatment of such non-motor symptoms in multiple scle rosis are still under *** studies for multiple sclerosis benefit from the use of disease-appropriate animal models,including experimental autoimmune *** to understanding the pathophysiology and developing treatments for non-motor symptoms,it is critical to chara cterize the animal model in terms of its ability to replicate certain non-motor features of multiple *** such,no single animal model can mimic the entire spectrum of *** review focuses on the non-motor symptoms that have been investigated in animal models of multiple sclerosis as well as possible underlying ***,we highlighted gaps in the literature to explain the nonmotor aspects of multiple sclerosis in expe rimental animal models,which will serve as the basis for future studies.

关键词： anxiety cognitive deficit depression experimental autoimmune encephalomyelitis motor disability neurological disorder pain pathophysiology preclinical study sensory impairments

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Nanobodies against African swine fever virus p72 and CD2v proteins as reagents for developing two cELISAs to detect viral antibodies

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Virologica Sinica 2024年第3期39卷 478-489页

作者： Jiahong Zhu Qingyuan Liu Liuya Li Runyu Zhang Yueting Chang Jiakai Zhao Siyu Liu Xinyu Zhao Xu Chen Yani Sun Qin Zhao department of Preventive veterinary Medicine College of Veterinary MedicineNorthwest A&F UniversityYangling Observing and Experimental Station of National Data Center of Animal HealthMinistry of AgricultureYangling712100China

African swine fever virus(ASFV)poses a significant threat to the global swine ***,there are no effective vaccines or treatments available to combat ASFV infection in *** primary means of controlling the spread of the disease is through rapid detection and subsequent elimination of infected ***,a lower virulent ASFV isolate with a deleted EP402R gene(CD2v-deleted)has been reported in China,which further complicates the control of ASFV infection in pig ***,an EP402R-deleted ASFV variant has been developed as a potential live attenuated vaccine candidate ***,it is crucial to develop detection methods that can distinguish wild-type and EP402R-deleted ASFV *** this study,two recombinant ASFV-p72 and-CD2v proteins were expressed using a prokaryotic system and used to immunize Bactrian ***,eight nanobodies against ASFV-p72 and ten nanobodies against ASFV-CD2v were *** the production of these nanobodies with horse radish peroxidase(HRP)fusion proteins,the ASFV-p72-Nb2-HRP and ASFV-CD2v-Nb22-HRP fusions were selected for the development of two competitive ELISAs(cELISAs)to detect anti-ASFV *** two cELISAs exhibited high sensitivity,good specificity,repeatability,and *** coincidence rate between the two cELISAs and commercial ELISA kits was 98.6%and 97.6%,***,the two cELISA for detecting antibodies against ASFV demonstrated ease of operation,a low cost,and a simple production *** two cELISAs could determine whether pigs were infected with wild-type or CD2v-deleted ASFV,and could play an important role in monitoring ASFV infections in pig farms.

关键词： African swine fever virus(ASFV) ASFV-p72 ASFV-CD2v Nanobody-HRP Competitive ELISA

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Changes in structural plasticity of hippocampal neurons in an animal model of multiple sclerosis

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Zoological research 2024年第2期45卷 398-414页

作者： Poornima D.E.Weerasinghe-Mudiyanselage Sohi Kang Joong-Sun Kim Sung-Ho Kim Hongbing Wang Taekyun Shin Changjong Moon department of veterinary Anatomy and Animal Behavior College of Veterinary Medicine and BK21 FOUR ProgramChonnam National UniversityGwangju 61186Republic of Korea department of Anatomy and Convergence Medical Science Institute of Health SciencesCollege of MedicineGyeongsang National UniversityJinju 52727Republic of Korea department of Physiology and Neuroscience Program Michigan State UniversityEast LansingMI 48824USA department of veterinary Anatomy College of Veterinary Medicine and Veterinary Medical Research InstituteJeju National UniversityJeju 63243Republic of Korea

Structural plasticity is critical for the functional diversity of neurons in the *** autoimmune encephalomyelitis(EAE)is the most commonly used model for multiple sclerosis(MS),successfully mimicking its key pathological features(inflammation,demyelination,axonal loss,and gliosis)and clinical symptoms(motor and non-motordysfunctions).Recentstudieshave demonstrated the importance of synaptic plasticity in EAE *** the present study,we investigated the features of behavioral alteration and hippocampal structural plasticity in EAE-affected mice in the early phase(11 days post-immunization,DPI)and chronic phase(28DPI).EAE-affected mice exhibited hippocampus-related behavioral dysfunction in the open field test during both early and chronic *** complexity was largely affected in the cornu ammonis 1(CA1)and CA3 apical and dentate gyrus(DG)subregions of the hippocampus during the chronic phase,while this effect was only noted in the CA1 apical subregion in the early ***,dendritic spine density was reduced in the hippocampal CA1 and CA3 apical/basal and DG subregions in the early phase of EAE,but only reduced in the DG subregion during the chronic ***,mRNA levels of proinflammatory cytokines(Il1β,Tnfα,and Ifnγ)and glial cell markers(Gfap and Cd68)were significantly increased,whereas the expression of activity-regulated cytoskeletonassociated protein(ARC)was reduced during the chronic ***,exposure to the aforementioned cytokines in primary cultures of hippocampal neurons reduced dendritic complexity and ARC *** cultures of hippocampal neurons also showed significantly reduced extracellular signal-regulated kinase(ERK)phosphorylation upon treatment with proinflammatory ***,these results suggest that autoimmune neuroinflammation alters structural plasticity in the hippocampus,possibly through the ERK-ARC pathway,indicating that this alteration may be associated with hi

关键词： Activity-regulated cytoskeleton-associated protein Anxiety-like behavior Experimental autoimmune encephalomyelitis Hippocampal dysfunction Neuroinflammation

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Enterogenic Stenotrophomonas maltophilia migrates to the mammary gland to induce mastitis by activating the calcium-ROS-AMPK-mTOR-autophagy pathway

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Journal of Animal Science and Biotechnology 2024年第1期15卷 236-252页

作者： Zhaoqi He Caijun Zhao Yuhong He Zhuoyu Liu Guyue Fan Kun Zhu Yiqi Wang Naisheng Zhang Yunhe Fu Xiaoyu Hu department of Clinical veterinary Medicine College of Veterinary MedicineJilin UniversityChangchun 130062Jilin ProvinceChina

Background Mastitis is an inflammatory disease of the mammary gland that has serious economic impacts on the dairy industry and endangers food *** previous study found that the body has a gut/rumen-mammary gland axis and that disturbance of the gut/rumen microbiota could result in‘gastroenterogenic mastitis'.However,the mechanism has not been fully ***,we found that long-term feeding of a high-concentrate diet induced mastitis in dairy cows,and the abundance of Stenotrophomonas maltophilia(***)was significantly increased in both the rumen and milk ***,we hypothesized that‘gastroenterogenic mastitis'can be induced by the migration of endogenous gut bacteria to the mammary ***,this study investigated the mechanism by which enterogenic *** induces *** First,*** was labelled with superfolder GFP and administered to mice via *** results showed that treatment with *** promoted the occurrence of mastitis and increased the permeability of the blood-milk barrier,leading to intestinal inflammation and intestinal ***,tracking of ingested *** revealed that *** could migrate from the gut to the mammary gland and induce ***,mammary gland transcriptome analysis showed that the calcium and AMPK signalling pathways were significantly upregulated in mice treated with ***,using mouse mammary epithelial cells(MMECs),we verified that *** induces mastitis through activation of the calcium-ROS-AMPK-mTOR-autophagy *** In conclusion,the results showed that enterogenic *** could migrate from the gut to the mammary gland via the gut-mammary axis and activate the calcium-ROS-AMPK-mTOR-autophagy pathway to induce *** the gut-mammary gland axis may also be an effective method to treat mastitis.

关键词： Calcium-ROS-AMPK-mTOR-autophagy pathway Gut-mammary axis Mastitis S.maltophilia

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The virulence regulator AbsR in avian pathogenic Escherichia coli has pleiotropic effects on bacterial physiology

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Journal of Integrative Agriculture 2024年第2期23卷 649-668页

作者： Dongfang Zhao Haobo Zhang Xinyang Zhang Fengwei Jiang Yijing Li Wentong Cai Ganwu Li State Key Laboratory for Animal Disease Control and Prevention Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069China College of veterinary Medicine Northeast Agricultural UniversityHarbin 150038China National Animal Tuberculosis Reference Laboratory Division of Zoonoses SurveillanceChina Animal Health and Epidemiology CenterQingdao 266032China department of veterinary Diagnostic and Production Animal Medicine College of Veterinary MedicineIowa State UniversityAmesIowa 50011-1134the United States

Avian pathogenic Escherichia coli(APEC)belonging to extraintestinal pathogenic ***(ExPEC)can cause severe infections in extraintestinal tissues in birds and humans,such as the lungs and ***(microcin production regulation,locus A,herein renamed AbsR,a blood survival regulator),a member of the MarR(multiple antibiotic resistance regulator)transcriptional regulator family,governs the expression of capsule biosynthetic genes in human ExPEC and represents a promising druggable target for ***,a deep understanding of the AbsR regulatory mechanism as well as its regulon is *** this study,we present a systems-level analysis of the APEC AbsR regulon using ChIP-Seq(chromatin immunoprecipitation sequencing)and RNA-Seq(RNA sequencing)*** found that AbsR directly regulates 99 genes and indirectly regulates 667 ***,we showed that:1)AbsR contributes to antiphagocytotic effects by macrophages and virulence in a mouse model for systemic infection by directly activating the capsular gene cluster;2)AbsR positively impacts biofilm formation via direct regulation of the T2SS(type II secretion system)but plays a marginal role in virulence;and 3)AbsR directly upregulates the acid tolerance signaling system EvgAS to withstand acid stress but is dispensable in ExPEC ***,our data indicate that the role of AbsR in virulence gene regulation is relatively conserved in ExPEC ***,this study provides a comprehensive analysis of the AbsR regulon and regulatory mechanism,and our data suggest that AbsR likely influences virulence primarily through the control of capsule ***,we found that AbsR severely represses the expression of the type I-F CRISPR(clustered regularly interspaced short palindromic repeats)-Cas(CRISPR associated)systems,which could have implications in CRISPR biology and application.

关键词： avian pathogenic Escherichia coli(APEC) extraintestinal pathogenic Escherichia coli(ExPEC) AbsR RNA-Seq ChIP-Seq gene regulation

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Hepatoprotective effect of Holothuria leucospilota methanolic extract on dimethyl nitrosamine-induced hepatotoxicity in rats

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Animal Models and Experimental Medicine 2024年第4期7卷 544-552页

作者： Fatemeh Dejan Amineh Daneshi Javad Rajabi Aslani Nasrollah Ahmadi Narges Eskandari Roozbahani Elaham Rahmanian Reza Behmanesh Hamid Reza Gheisari department of Basic Sciences School of Veterinary MedicineShiraz UniversityShirazIran department of Education Farzanegan High SchoolTehranKohgiluyeh and Boyer AhmadIran Food Hygiene department School of Veterinary MedicineShiraz UniversityShirazIran department of Pathology School of Veterinary MedicineShiraz UniversityShirazIran Clinical research Development center Imam Reza HospitalKermanshah University of Medical SciencesKermanshahIran Molecular Dermatology research center Shiraz University of Medical SciencesShirazIran department of Anatomy School of MedicineShiraz University of Medical SciencesShirazIran Faculty of veterinary Medicine Garmsar BranchDepartment of RadiologyIslamic Azad UniversityGarmsarIran

Background:Complementary medicine is an interesting field for extracting bio-active compounds from various plant and animal *** hepatoprotective effect of the methanolic extract of a species of sea cucumber called Holothuria leu-cospilota in an animal model of liver cancer caused by dimethyl nitrosamine(DMN)was ***:Wistar female rats were randomly divided into five groups(n=12):control(intact),positive control(received 1%DMN[10 mg/kg/week,intraperitoneally]for 12 weeks),and three treatment groups(received 50,100,and 200 mg/kg/day ***-cospilota extract orally for 12 weeks along with intraperitoneal administration of 1%DMN[10 mg/kg/week]).In all groups,ultrasound was performed on the liver every week to check its *** sampling and liver isolation were performed on three occasions,at 4,8,and 12 weeks,to check liver enzymes and the histopathological condition of the liver tissue(every week,four animals from each group were randomly selected).Results:Liver density changes were evident from the eighth week onward in the positive control *** results indicated pathologic changes in the positive control group after 4 *** increase in liver enzymes in the posi-tive control group was significantly different from that in the treatment and control ***:We demonstrated the hepatoprotective effect of *** on DMN-induced liver damage in rats using biochemical and histological parameters and *** additional research(in silico or in vitro)is needed to find the exact mechanism and the main biological compound in ***.

关键词： dimethyl nitrosamine hepatotoxicity Holothuria leucospilota sea cucumber

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Ancylostoma ceylanicum and other zoonotic canine hookworms:neglected public and animal health risks in the Asia-Pacific region

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Animal Diseases 2024年第1期4卷 59-74页

作者： Jan Clyden B.Tenorio lan Kim B.Tabios Tawin Inpankaew Adrian P.Ybanez Saruda Tiwananthagorn Sirikachorn Tangkawattana Sutas Suttiprapa Tropical Medicine Graduate Program Faculty of MedicineKhon Kaen UniversityKhon Kaen 40002Thailand department of Tropical Medicine Faculty of MedicineKhon Kaen UniversityKhon Kaen 40002Thailand WHO Collaborating center for research and Control of Opisthorchiasis(Southeast Asian Liver Fluke Disease) Tropical Disease Research CenterKhon Kaen UniversityKhon Kaen 40002Thailand department of veterinary Paraclinical Sciences College of Veterinary MedicineUniversity of Southern MindanaoKabacan9407 CotabatoPhilippines Institute of Biology College of ScienceUniversity of the Philippines DilimanManila 1101Philippines department of Parasitology Faculty of Veterinary MedicineKasetsart UniversityBangkok 10900Thailand College of veterinary Medicine at Barili Campus and center for Vector-Borne and Protozoan Diseases at Main Campus Cebu Technological UniversityCebu City 6000Philippines department of veterinary Biosciences and Public Health Faculty of Veterinary MedicineChiang Mai UniversityChiang Mai 50100Thailand research center for veterinary Biosciences and veterinary Public Health Chiang Mai UniversityChiang Mai 50100Thailand Faculty of veterinary Medicine Khon Kaen University.Khon Kaen 40002Thailand

Zoonotic hookworm infections remain a significant public health problem,causing nearly 500 milion cases globally and approximately four million disability-adjusted life years lost *** than one-fth of these cases are attrib-uted to Ancylostoma ceylanicum,an emerging zoonotic health issue in the Asia-Pacific *** review presents key research gaps regarding the epidemiology,diagnosis,control,prevention and elimination of *** and other canine zoonotic hookworms as neglected health *** is the second most prevalent human hook-worm in the region;it is the most common hookworm among dogs and cats-reservoirs of zoonotic *** population genetic and phylogenetic analyses revealed that *** has three possible transmis-sion dynamics:zoonotic,animal-only,and human-only *** actual burden of zoonotic ancylostomiasis in most endemic countries remains unknown due to the use of parasitological techniques(e.g.,Kato-Katz thick smear and floatation techniques)that have reduced diagnostic performance and do not allow accurate species identifica-tion in helminth *** emergence of benzimidazole resistance in soil-transmitted helminths(STHs),includ-ing hookworms,is a concern due to the protracted implementation of mass drug administration(MDA).Resistance is conferred by single nucleotide polymorphisms(SNPs)that occur in theβ-tubulin isotype 1 *** mutations have been reported in drug-resistant *** but have not been found in *** in the *** remains understudied in the Asia-Pacific *** zoonotic nature of the parasite warrants investigation of its occur-rence in human and animal reservoir hosts to understand the dynamics of zoonotic transmission in different endemic *** detection of benzimidazole resistance-associated SNPs in zoonotic hookworms from Asia-Pacific countries has yet to be thoroughly *** the high level of hookworm endemicity in the region

关键词： Soil-transmitted helminths Neglected tropical diseases Mass drug administration Deworming veterinary public health

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Additively manufactured Ti–Ta–Cu alloys for the next-generation load-bearing implants

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International Journal of Extreme Manufacturing 2024年第1期6卷 353-374页

作者： Amit Bandyopadhyay Indranath Mitra Sushant Ciliveri Jose D Avila William Dernell Stuart B Goodman Susmita Bose W.M.Keck Biomedical Materials research Laboratory School of Mechanical and Materials EngineeringWashington State UniversityPullmanWA 99164United States of America veterinary Clinical Sciences College of Veterinary MedicineWashington State UniversityPullmanWA 99164United States of America department of Orthopedic Surgery Stanford University Medical CenterRedwood CityCA 94063United States of America

Bacterial colonization of orthopedic implants is one of the leading causes of failure and clinical complexities for load-bearing metallic implants. Topical or systemic administration of antibiotics may not offer the most efficient defense against colonization, especially in the case of secondary infection, leading to surgical removal of implants and in some cases even limbs. In this study, laser powder bed fusion was implemented to fabricate Ti3Al2V alloy by a 1:1 weight mixture of CpTi and Ti6Al4V powders. Ti-Tantalum(Ta)–Copper(Cu) alloys were further analyzed by the addition of Ta and Cu into the Ti3Al2V custom alloy. The biological,mechanical, and tribo-biocorrosion properties of Ti3Al2V alloy were evaluated. A 10 wt.% Ta(10Ta) and 3 wt.% Cu(3Cu) were added to the Ti3Al2V alloy to enhance biocompatibility and impart inherent bacterial resistance. Additively manufactured implants were investigated for resistance against Pseudomonas aeruginosa and Staphylococcus aureus strains of bacteria for up to 48 h. A 3 wt.% Cu addition to Ti3Al2V displayed improved antibacterial efficacy, i.e.78%–86% with respect to CpTi. Mechanical properties for Ti3Al2V–10Ta–3Cu alloy were evaluated, demonstrating excellent fatigue resistance, exceptional shear strength, and improved tribological and tribo-biocorrosion characteristics when compared to Ti6Al4V. In vivo studies using a rat distal femur model revealed improved early-stage osseointegration for alloys with10 wt.% Ta addition compared to CpTi and Ti6Al4V. The 3 wt.% Cu-added compositions displayed biocompatibility and no adverse infammatory response in vivo. Our results establish the Ti3Al2V–10Ta–3Cu alloy’s synergistic effect on improving both in vivo biocompatibility and microbial resistance for the next generation of load-bearing metallic implants.

关键词： Ti6Al4V load-bearing implants additive manufacturing 3D printing antibacterial performance

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Effects of Moringa oleifera aqueous extract on the growth performance,blood characteristics,and histological features of gills and livers in Nile tilapia

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Aquaculture and Fisheries 2024年第1期9卷 85-92页

作者： Mahamoud A.Emam Ramy M.Shourbela Waleed N.El-Hawarry Seham Y.Abo-Kora Fatma Abdel-Monem Gad Ashraf M.Abd El-latif Mahmoud A.O.Dawood Histology department Faculty of Veterinary MedicineBenha UniversityBenha13511Egypt department of Animal Husbandry and Animal Wealth Development Faculty of Veterinary MedicineAlexandria UniversityAlexandria22758Egypt Pharmacology department Faculty of Veterinary MedicineBenha UniversityBenha13511Egypt Clinical Pathology department Faculty of Veterinary MedicineBenha UniversityBenha13511Egypt department of Aquatic Animal Diseases and Management Faculty of Veterinary MedicineBenha UniversityBenha13511Egypt Animal Production department Faculty of AgricultureKafrelsheikh UniversityKafr El-Sheikh33511Egypt The center for Applied research on the Environment and Sustainability The American University in CairoCairo11835Egypt

Moringa oleifera is well known as a highly nutritious plant and a water purifier in fish *** fish has many impressive qualities making them very suitable for ***,this study aimed to evaluate the effect of supplementation of Moringa oleifera aqueous extract(MOAE)at varying concentrations(0,100,200,and 400 mg/kg diet for 90 days)on growth performance,hematological,and some biochemical parameters of Nile ***,histological changes and immunohistochemical expression of caspase 3 in gills and livers were *** enhancements(P<0.05)in the final weight,weight gain,specific growth rate,red blood cells,hemoglobin,packed cell volume,white blood cells,and total serum protein were observed in fish fed 200 mg MOAE/kg diet compared with other ***,a significant reduction(P<0.05)of liver and kidney function tests and non-significant changes in lipid profile were *** the other hand,fish fed 400 mg MOAE/kg diet revealed disturbance in the corresponding *** histological degenerative changes were detected in the gills and liver of fish fed 400 mg MOAE/kg *** comparison,fish fed 200 mg MOAE/kg diet showed the best histo-architectures of these organs compared with other *** immune expressions of caspase 3 were determined where gills and liver of fish fed 400 mg MOAE/kg diet revealed extensive number of positive caspase 3 cells indicating severe apoptotic ***,a dietary MOAE at 200 mg/kg diet is recommended to improve the growth performance,hematological parameters,liver and kidney functions in Nile ***,caution must be considered when using MOAE at a concentration over 200 mg/kg diet.

关键词： Growth performance Liver Gills Moringa oleifera Nile tilapia

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