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Lv Y, Gao X, Dai Q, Zhu L, Liu S, Hu Z, Lu J, Zhou H, Jin J, Mei Z. Functional insights of digestion, absorption, and immunity in different segments of the intestine in Hemibarbus labeo from transcriptomic analysis. COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY. PART D, GENOMICS & PROTEOMICS 2024; 52:101278. [PMID: 38906043 DOI: 10.1016/j.cbd.2024.101278] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/13/2024] [Revised: 05/26/2024] [Accepted: 06/12/2024] [Indexed: 06/23/2024]
Abstract
The intestine is an important organ for food digestion and absorption and body immunity in fish. In this study, we investigated the abundance of transcripts from different segments of the intestinal tract using transcriptome sequencing technology in Hemibarbus labeo, to provide functional insights into digestion, absorption, and immunity in the anterior intestine (AI), middle intestine (MI), and posterior intestine (PI). We found 5646 differentially expressed genes (DEGs), which were significantly enriched to GO terms of carbohydrate metabolic process, transmembrane transport, iron ion binding, lipid metabolic process, and KEGG pathway of fat digestion and absorption, mineral absorption, protein digestion and absorption, vitamin digestion and absorption, indicating that the digestion and absorption function of food is different in AI, MI, and PI. In practice, most genes, enriched in the KEGG pathway for digestion and absorption of nutrients, are upregulated in AI and MI, indicating stronger roles for food digestion and absorption in these segments. Furthermore, we found that genes involved in the KEGG pathway of lysosome and endocytosis pathway are upregulated in PI, suggesting stronger antigen-presenting capabilities in PI. However, some cytokine receptor genes, including ccr4, cxcr2, tnfrsf9, il6r, csf3r, and cxcr4, are highly expressed in AI, reflecting the regional immune specialization in different segments. This study provides functional insights into digestion, absorption, and immunity in different segments of the intestine and supports the regional functional specialization within different segments of the intestine in H. labeo.
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Affiliation(s)
- Yaoping Lv
- College of Ecology, Lishui University, Lishui 323000, Zhejiang, China
| | - Xinming Gao
- College of Ecology, Lishui University, Lishui 323000, Zhejiang, China.
| | - Qingmin Dai
- College of Ecology, Lishui University, Lishui 323000, Zhejiang, China
| | - Ling Zhu
- College of Ecology, Lishui University, Lishui 323000, Zhejiang, China
| | - Siqi Liu
- College of Ecology, Lishui University, Lishui 323000, Zhejiang, China
| | - Zehui Hu
- Zhejiang Marine Fisheries Research Institute, Zhoushan 316100, Zhejiang, China
| | - Junkai Lu
- Cixi Fisheries Technology Extension Center, Ningbo 315300, Zhejiang, China
| | - Haidong Zhou
- Suichang Fisheries and Agricultural Machinery Technology Extension Station, Lishui 323399, Zhejiang, China
| | - Jing Jin
- Zhejiang Fisheries Technology Extension Center, Hangzhou 311100, Zhejiang, China
| | - Zufei Mei
- Jinman Aquatic Seedling Farm, Lishui 323006, Zhejiang, China
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Zhang L, Liu H, Zhang H, Yuan H, Ren D. Lemairamin (Wgx-50) Attenuates DSS-Induced Intestinal Inflammation in Zebrafish. Int J Mol Sci 2024; 25:9510. [PMID: 39273457 PMCID: PMC11395399 DOI: 10.3390/ijms25179510] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/31/2024] [Revised: 08/23/2024] [Accepted: 08/29/2024] [Indexed: 09/15/2024] Open
Abstract
Inflammatory bowel disease (IBD) is a chronic non-specific intestinal inflammatory disease that affects millions of people worldwide, and current treatment methods have certain limitations. This study aimed to explore the therapeutic potential and mechanism of action of lemairamin (Wgx-50) in inflammatory bowel disease (IBD). We used dextran sulfate sodium (DSS)-treated zebrafish as an inflammatory bowel disease model, and observed the effect of Wgx-50 on DSS-induced colitis inflammation. The results of the study showed that Wgx-50 could reduce the expression of pro-inflammatory cytokines induced by DSS and inhibit the recruitment of neutrophils to the site of intestinal injury. Further experiments revealed that Wgx-50 exerted its anti-inflammatory effect by regulating the activation of the Akt pathway. These research findings indicate that Wgx-50 possesses anti-inflammatory activity.
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Affiliation(s)
- Ling Zhang
- College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China
| | - Huiru Liu
- College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China
| | - Haoyi Zhang
- College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China
| | - Hao Yuan
- College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China
| | - Dalong Ren
- College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China
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Yang J, Isaka T, Kikuchi K, Numayama-Tsuruta K, Ishikawa T. Bacterial accumulation in intestinal folds induced by physical and biological factors. BMC Biol 2024; 22:76. [PMID: 38581018 PMCID: PMC10998401 DOI: 10.1186/s12915-024-01874-5] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/15/2023] [Accepted: 03/25/2024] [Indexed: 04/07/2024] Open
Abstract
BACKGROUND The gut microbiota, vital for host health, influences metabolism, immune function, and development. Understanding the dynamic processes of bacterial accumulation within the gut is crucial, as it is closely related to immune responses, antibiotic resistance, and colorectal cancer. We investigated Escherichia coli behavior and distribution in zebrafish larval intestines, focusing on the gut microenvironment. RESULTS We discovered that E. coli spread was considerably suppressed within the intestinal folds, leading to a strong physical accumulation in the folds. Moreover, a higher concentration of E. coli on the dorsal side than on the ventral side was observed. Our in vitro microfluidic experiments and theoretical analysis revealed that the overall distribution of E. coli in the intestines was established by a combination of physical factor and bacterial taxis. CONCLUSIONS Our findings provide valuable insight into how the intestinal microenvironment affects bacterial motility and accumulation, enhancing our understanding of the behavioral and ecological dynamics of the intestinal microbiota.
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Affiliation(s)
- Jinyou Yang
- School of Intelligent Medicine, China Medical University, Shenyang, 110122, China.
| | - Toma Isaka
- Department of Biomedical Engineering, Graduate School of Biomedical Engineering, Tohoku University, 6-6-01 Aoba, Sendai, 980-8579, Japan
| | - Kenji Kikuchi
- Department of Biomedical Engineering, Graduate School of Biomedical Engineering, Tohoku University, 6-6-01 Aoba, Sendai, 980-8579, Japan
- Department of Finemechanics, Graduate School of Engineering, Tohoku University, 6-6-01 Aoba, Sendai, 980-8579, Japan
| | - Keiko Numayama-Tsuruta
- Department of Biomedical Engineering, Graduate School of Biomedical Engineering, Tohoku University, 6-6-01 Aoba, Sendai, 980-8579, Japan
| | - Takuji Ishikawa
- Department of Biomedical Engineering, Graduate School of Biomedical Engineering, Tohoku University, 6-6-01 Aoba, Sendai, 980-8579, Japan
- Department of Finemechanics, Graduate School of Engineering, Tohoku University, 6-6-01 Aoba, Sendai, 980-8579, Japan
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Sanahuja I, Ruiz A, Firmino JP, Reyes-López FE, Ortiz-Delgado JB, Vallejos-Vidal E, Tort L, Tovar-Ramírez D, Cerezo IM, Moriñigo MA, Sarasquete C, Gisbert E. Debaryomyces hansenii supplementation in low fish meal diets promotes growth, modulates microbiota and enhances intestinal condition in juvenile marine fish. J Anim Sci Biotechnol 2023; 14:90. [PMID: 37422657 DOI: 10.1186/s40104-023-00895-4] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/20/2023] [Accepted: 05/11/2023] [Indexed: 07/10/2023] Open
Abstract
BACKGROUND The development of a sustainable business model with social acceptance, makes necessary to develop new strategies to guarantee the growth, health, and well-being of farmed animals. Debaryomyces hansenii is a yeast species that can be used as a probiotic in aquaculture due to its capacity to i) promote cell proliferation and differentiation, ii) have immunostimulatory effects, iii) modulate gut microbiota, and/or iv) enhance the digestive function. To provide inside into the effects of D. hansenii on juveniles of gilthead seabream (Sparus aurata) condition, we integrated the evaluation of the main key performance indicators coupled with the integrative analysis of the intestine condition, through histological and microbiota state, and its transcriptomic profiling. RESULTS After 70 days of a nutritional trial in which a diet with low levels of fishmeal (7%) was supplemented with 1.1% of D. hansenii (17.2 × 105 CFU), an increase of ca. 12% in somatic growth was observed together with an improvement in feed conversion in fish fed a yeast-supplemented diet. In terms of intestinal condition, this probiotic modulated gut microbiota without affecting the intestine cell organization, whereas an increase in the staining intensity of mucins rich in carboxylated and weakly sulphated glycoconjugates coupled with changes in the affinity for certain lectins were noted in goblet cells. Changes in microbiota were characterized by the reduction in abundance of several groups of Proteobacteria, especially those characterized as opportunistic groups. The microarrays-based transcriptomic analysis found 232 differential expressed genes in the anterior-mid intestine of S. aurata, that were mostly related to metabolic, antioxidant, immune, and symbiotic processes. CONCLUSIONS Dietary administration of D. hansenii enhanced somatic growth and improved feed efficiency parameters, results that were coupled to an improvement of intestinal condition as histochemical and transcriptomic tools indicated. This probiotic yeast stimulated host-microbiota interactions without altering the intestinal cell organization nor generating dysbiosis, which demonstrated its safety as a feed additive. At the transcriptomic level, D. hansenii promoted metabolic pathways, mainly protein-related, sphingolipid, and thymidylate pathways, in addition to enhance antioxidant-related intestinal mechanisms, and to regulate sentinel immune processes, potentiating the defensive capacity meanwhile maintaining the homeostatic status of the intestine.
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Affiliation(s)
- Ignasi Sanahuja
- Aquaculture Program, Institute of Agrifood Research and Technology (IRTA), La Ràpita, 43540, Spain
| | - Alberto Ruiz
- Aquaculture Program, Institute of Agrifood Research and Technology (IRTA), La Ràpita, 43540, Spain
| | - Joana P Firmino
- Aquaculture Program, Institute of Agrifood Research and Technology (IRTA), La Ràpita, 43540, Spain
| | - Felipe E Reyes-López
- Centro de Biotecnología Acuícola, Universidad de Santiago de Chile, Santiago, Chile
| | - Juan B Ortiz-Delgado
- Instituto de Ciencias Marinas de Andalucía (ICMAN-CSIC), Avda. República Saharaui nº 2, Campus Universitario Río San Pedro, Puerto Real, Cádiz, 11510, Spain
| | - Eva Vallejos-Vidal
- Núcleo de Investigaciones Aplicadas en Ciencias Veterinarias y Agronómicas, Facultad de Medicina Veterinaria y Agronomía, Universidad de Las Américas, Santiago, Chile
| | - Lluis Tort
- Department of Cell Biology, Physiology, and Immunology, Universitat Autonoma de Barcelona, Barcelona, Spain
| | | | - Isabel M Cerezo
- Department of Microbiology, Instituto de Biotecnología Y Desarrollo Azul (IBYDA), Faculty of Sciences, University of Malaga, 29010, Malaga, Spain
- SCBI, Bioinformatic Unit, University of Malaga, 29590, Malaga, Spain
| | - Miguel A Moriñigo
- Department of Microbiology, Instituto de Biotecnología Y Desarrollo Azul (IBYDA), Faculty of Sciences, University of Malaga, 29010, Malaga, Spain
| | - Carmen Sarasquete
- Instituto de Ciencias Marinas de Andalucía (ICMAN-CSIC), Avda. República Saharaui nº 2, Campus Universitario Río San Pedro, Puerto Real, Cádiz, 11510, Spain
| | - Enric Gisbert
- Aquaculture Program, Institute of Agrifood Research and Technology (IRTA), La Ràpita, 43540, Spain.
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Kuang Y, Guo H, Ouyang K, Wang X, Li D, Li L. Nano-TiO 2 aggravates immunotoxic effects of chronic ammonia stress in zebrafish (Danio rerio) intestine. Comp Biochem Physiol C Toxicol Pharmacol 2023; 266:109548. [PMID: 36626958 DOI: 10.1016/j.cbpc.2023.109548] [Citation(s) in RCA: 4] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/24/2022] [Revised: 12/29/2022] [Accepted: 01/04/2023] [Indexed: 01/09/2023]
Abstract
Ammonia and nano-TiO2 are commonly found pollutants in aquatic environments around the world. NH3 has been proved to be absorbed on nano-TiO2 surface, therefore, the biosafety and environmental effects of ammonia and co-occurring nano-TiO2 in aquatic environments has increased considerably in recent years. To explore the potential interactive effects and mechanisms of ammonia and nano-TiO2 on the intestinal immune system, three-month-old female zebrafish were exposed to total ammonia nitrogen (TAN; 0, 3, 30 mg/L) with or without nano-TiO2 (1 mg/L) for 60 d. The results showed that intestinal ammonia levels increased with the increase of TAN exposure concentration in the presence of nano-TiO2. Histopathological analysis demonstrated that both TAN and nano-TiO2 caused cell vacuolation, lymphocyte infiltration and goblet cells hyperplasia in the intestine mucosa. Our study also found that the contents and gene expression levels of lysozyme (lys) and β-defensin (def-β) in the intestine of zebrafish exposed to TAN alone or combined with nano-TiO2 were significantly reduced, suggesting a decline in the intestinal innate immunity of fish. A broad upregulation of TLRs-related genes indicated that TAN and nano-TiO2 could activate TLR4/5-mediated MyD88-dependent pathway, and eventually induce intestinal inflammation. It should be noted that TAN combined with nano-TiO2 had more significant inhibitory effects on the intestinal structure and innate immune responses than TAN alone. Current data suggested that ammonia and nano-TiO2 had a synergistic inhibitory effect on intestinal mucosal immunity, and their associated health risk to aquatic animals and the water ecosystem should not be underestimated.
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Affiliation(s)
- Yu Kuang
- College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China
| | - Honghui Guo
- College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China
| | - Kang Ouyang
- College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China
| | - Xinyu Wang
- College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China
| | - Dapeng Li
- College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China; Engineering Research Center of Green development for Conventional Aquatic Biological Industry in the Yangtze River Economic Belt, Ministry of Education, Wuhan 430070, China; Hubei Provincial Engineering Laboratory for Pond Aquaculture, Wuhan 430070, China; Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan 430070, China
| | - Li Li
- College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China; Engineering Research Center of Green development for Conventional Aquatic Biological Industry in the Yangtze River Economic Belt, Ministry of Education, Wuhan 430070, China; Hubei Provincial Engineering Laboratory for Pond Aquaculture, Wuhan 430070, China; Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan 430070, China.
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6
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Stosik M, Tokarz-Deptuła B, Deptuła W. Immunity of the intestinal mucosa in teleost fish. FISH & SHELLFISH IMMUNOLOGY 2023; 133:108572. [PMID: 36717066 DOI: 10.1016/j.fsi.2023.108572] [Citation(s) in RCA: 14] [Impact Index Per Article: 7.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/25/2022] [Revised: 01/26/2023] [Accepted: 01/26/2023] [Indexed: 06/18/2023]
Abstract
The paper presents the problem of intestinal mucosa immunity in teleost fish. The immunity of the intestinal mucosa in teleost fish depends on the elements and mechanisms with different organizational/structural and functional properties than in mammals. The organization of the elements of intestinal mucosal immunitya in these animals is associated with the presence of immune cells that fulfil the functions assigned to the induction and effector sites of mucosal immunity in mammals; they are located at various histological sites of the mucosa - in the lamina propria (LP) and in the surface epithelium. The presence of mucosa-associated lymphoid tissue (MALT) has not been demonstrated in teleost fish, and the terminology used in relation to the structure and function of the mucosa immunity components in teleost fish is inadequate. In this article, we review the knowledge of intestinal mucosal immunity in teleost fish, with great potential for knowledge and practical applications especially in the field of epidemiological safety. We discuss the organization and functional properties of the elements that determine this immunity, according to current data and taking into account the tissue definition and terminology adopted by the Society for Mucosal Immunology General Assembly (13th ICMI in Tokyo, 2007).
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Affiliation(s)
- Michał Stosik
- Institute of Biological Sciences, University of Zielona Góra, Poland
| | | | - Wiesław Deptuła
- Institute of Veterinary Medicine, Faculty of Biological and Veterinary Sciences, Nicolaus Copernicus University in Toruń, Poland
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Xia H, Chen H, Cheng X, Yin M, Yao X, Ma J, Huang M, Chen G, Liu H. Zebrafish: an efficient vertebrate model for understanding role of gut microbiota. Mol Med 2022; 28:161. [PMID: 36564702 PMCID: PMC9789649 DOI: 10.1186/s10020-022-00579-1] [Citation(s) in RCA: 33] [Impact Index Per Article: 11.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/12/2022] [Accepted: 11/21/2022] [Indexed: 12/24/2022] Open
Abstract
Gut microbiota plays a critical role in the maintenance of host health. As a low-cost and genetically tractable vertebrate model, zebrafish have been widely used for biological research. Zebrafish and humans share some similarities in intestinal physiology and function, and this allows zebrafish to be a surrogate model for investigating the crosstalk between the gut microbiota and host. Especially, zebrafish have features such as high fecundity, external fertilization, and early optical transparency. These enable the researchers to employ the fish to address questions not easily addressed in other animal models. In this review, we described the intestine structure of zebrafish. Also, we summarized the methods of generating a gnotobiotic zebrafish model, the factors affecting its intestinal flora, and the study progress of gut microbiota functions in zebrafish. Finally, we discussed the limitations and challenges of the zebrafish model for gut microbiota studies. In summary, this review established that zebrafish is an attractive research tool to understand mechanistic insights into host-microbe interaction.
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Affiliation(s)
- Hui Xia
- College of Basic Medicine, Hubei University of Chinese Medicine, Huangjiahu West Road 16, Hongshan Disctrict, Wuhan, 430065, China
| | - Huimin Chen
- College of Basic Medicine, Hubei University of Chinese Medicine, Huangjiahu West Road 16, Hongshan Disctrict, Wuhan, 430065, China
| | - Xue Cheng
- College of Basic Medicine, Hubei University of Chinese Medicine, Huangjiahu West Road 16, Hongshan Disctrict, Wuhan, 430065, China
| | - Mingzhu Yin
- College of Basic Medicine, Hubei University of Chinese Medicine, Huangjiahu West Road 16, Hongshan Disctrict, Wuhan, 430065, China
| | - Xiaowei Yao
- College of Basic Medicine, Hubei University of Chinese Medicine, Huangjiahu West Road 16, Hongshan Disctrict, Wuhan, 430065, China
| | - Jun Ma
- College of Basic Medicine, Hubei University of Chinese Medicine, Huangjiahu West Road 16, Hongshan Disctrict, Wuhan, 430065, China
| | - Mengzhen Huang
- College of Basic Medicine, Hubei University of Chinese Medicine, Huangjiahu West Road 16, Hongshan Disctrict, Wuhan, 430065, China
| | - Gang Chen
- Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan, 430061, China.
| | - Hongtao Liu
- College of Basic Medicine, Hubei University of Chinese Medicine, Huangjiahu West Road 16, Hongshan Disctrict, Wuhan, 430065, China.
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8
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Nikiforov-Nikishin A, Smorodinskaya S, Kochetkov N, Nikiforov-Nikishin D, Danilenko V, Bugaev O, Vatlin A, Abrosimova N, Antipov S, Kudryavtsev A, Klimov V. Effects of Three Feed Additives on the Culturable Microbiota Composition and Histology of the Anterior and Posterior Intestines of Zebrafish ( Danio rerio). Animals (Basel) 2022; 12:2424. [PMID: 36139282 PMCID: PMC9495144 DOI: 10.3390/ani12182424] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/15/2022] [Revised: 09/06/2022] [Accepted: 09/14/2022] [Indexed: 12/03/2022] Open
Abstract
In this study, the effect of three promising feed additives (chelated compounds of trace elements, butyric acid, lycopene) on changes in the culturable microbiota and histological parameters of two sections of the intestines of Danio rerio (zebrafish) was studied. The use of these feed additives can help to eliminate the deficiency of trace elements, modulate the composition of the microbiota due to the postbiotic properties of butyric acid, and reduce oxidative stress when using lycopene. Incorporation of the investigated supplements in the feed resulted in a significant change in the relative abundance of certain groups of microorganisms. The taxonomic diversity of cultured microorganisms did not differ in the anterior and posterior intestines, while there were differences in the relative abundance of these microorganisms. The most sensitive groups of microorganisms were the genera Bacillus and Serratia. A significant effect on the composition of the cultured microbiota was caused by lycopene (in all studied concentrations), leading to a significant increase in the relative abundance of Firmicutes in the anterior gut. Studies of the histological structure of the anterior and posterior guts have shown the relationship between the barrier and secretory functions of the gut and the composition of the microbiota while using butyric acid (1 and 2 g kg-1) and trace element chelated compounds (2 mg kg-1). This culture-dependent method of studying the microbiome makes it possible to assess changes in some representatives of the main groups of microorganisms (Firmicutes and Proteobacteria). Despite the incompleteness of the data obtained by the culture-dependent method, its application makes it possible to assess the bioactive properties of feed and feed additives and their impact on the microbiota involved in digestive processes.
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Affiliation(s)
- Alexei Nikiforov-Nikishin
- Faculty of Biotechnology and Fisheries, Moscow State University of Technologies and Management (FCU), 73 Zemlyanoy Val Str., 109004 Moscow, Russia
| | - Svetlana Smorodinskaya
- Faculty of Biotechnology and Fisheries, Moscow State University of Technologies and Management (FCU), 73 Zemlyanoy Val Str., 109004 Moscow, Russia
| | - Nikita Kochetkov
- Faculty of Biotechnology and Fisheries, Moscow State University of Technologies and Management (FCU), 73 Zemlyanoy Val Str., 109004 Moscow, Russia
| | - Dmitry Nikiforov-Nikishin
- Faculty of Biotechnology and Fisheries, Moscow State University of Technologies and Management (FCU), 73 Zemlyanoy Val Str., 109004 Moscow, Russia
| | - Valery Danilenko
- Laboratory of Bacterial Genetics, Vavilov Institute of General Genetics, Russian Academy of Sciences, 119333 Moscow, Russia
| | - Oleg Bugaev
- Faculty of Biotechnology and Fisheries, Moscow State University of Technologies and Management (FCU), 73 Zemlyanoy Val Str., 109004 Moscow, Russia
| | - Aleksey Vatlin
- Laboratory of Bacterial Genetics, Vavilov Institute of General Genetics, Russian Academy of Sciences, 119333 Moscow, Russia
| | - Nina Abrosimova
- Department of Aquaculture Techniques, Don State Technical University, Gagarin Square 1, 344003 Rostov-on-Don, Russia
| | - Sergei Antipov
- Department of Biophysics and Biotechnology, Voronezh State University, University Square 1, 394063 Voronezh, Russia
| | - Alexander Kudryavtsev
- Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, Russia
| | - Viktor Klimov
- Faculty of Biotechnology and Fisheries, Moscow State University of Technologies and Management (FCU), 73 Zemlyanoy Val Str., 109004 Moscow, Russia
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9
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Hattab S, Dagher AM, Wheeler RT. Pseudomonas Synergizes with Fluconazole against Candida during Treatment of Polymicrobial Infection. Infect Immun 2022; 90:e0062621. [PMID: 35289633 PMCID: PMC9022521 DOI: 10.1128/iai.00626-21] [Citation(s) in RCA: 10] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/18/2021] [Accepted: 02/10/2022] [Indexed: 12/14/2022] Open
Abstract
Polymicrobial infections are challenging to treat because we don't fully understand how pathogens interact during infection and how these interactions affect drug efficacy. Candida albicans and Pseudomonas aeruginosa are opportunistic pathogens that can be found in similar sites of infection such as in burn wounds and most importantly in the lungs of CF and mechanically ventilated patients. C. albicans is particularly difficult to treat because of the paucity of antifungal agents, some of which lack fungicidal activity. In this study, we investigated the efficacy of anti-fungal treatment during C. albicans-P. aeruginosa coculture in vitro and co-infection in the mucosal zebrafish infection model analogous to the lung. We find that P. aeruginosa enhances the activity of fluconazole (FLC), an anti-fungal drug that is fungistatic in vitro, to promote both clearance of C. albicans during co-infection in vivo and fungal killing in vitro. This synergy between FLC treatment and bacterial antagonism is partly due to iron piracy, as it is reduced upon iron supplementation and knockout of bacterial siderophores. Our work demonstrates that FLC has enhanced activity in clinically relevant contexts and highlights the need to understand antimicrobial effectiveness in the complex environment of the host with its associated microbial communities.
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Affiliation(s)
- Siham Hattab
- Department of Molecular & Biomedical Sciences, University of Maine, Orono, Maine, USA
| | - Anna-Maria Dagher
- Department of Molecular & Biomedical Sciences, University of Maine, Orono, Maine, USA
| | - Robert T. Wheeler
- Department of Molecular & Biomedical Sciences, University of Maine, Orono, Maine, USA
- Graduate School of Biomedical Sciences and Engineering, University of Maine, Orono, Maine, USA
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10
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Das S, Pradhan C, Pillai D. β-Defensin: An adroit saviour in teleosts. FISH & SHELLFISH IMMUNOLOGY 2022; 123:417-430. [PMID: 35331882 DOI: 10.1016/j.fsi.2022.03.017] [Citation(s) in RCA: 16] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 09/10/2021] [Revised: 03/16/2022] [Accepted: 03/16/2022] [Indexed: 06/14/2023]
Abstract
β-Defensin (BD) is an important first line innate defense molecule with potent antimicrobial and immunomodulatory activities in fish. The signatures of β-defensins are the presence of a net cationic charge and three intramolecular disulfide bonds mediated by six conserved cysteines. It consists of three exons and two introns. The signal peptide is usually conserved and sequence divergence is mostly seen in mature peptide region. The diverse amino acid sequences of matured peptide contribute to a strong positive selection and broad-spectrum antimicrobial activity. It is constitutively expressed in both mucosal as well as systemic sites. Increased expression of β-defensin was mostly reported in bacterial and viral infections in fish. Its role during parasitic and fungal infections is yet to be investigated. β-Defensin isoforms such as BD-1, BD-2, BD-3, BD-4 and BD-5 can be witnessed even in early developmental days to different pathogenic exposure in fish. β-Defensins possess adjuvant properties to enhance antigen-specific immunity promoting both cellular and humoral immune response. It significantly reduces/increases bacterial colonization or viral copy numbers when overexpressed/knockdown. Based on its chemotactic and activating potentials, it can contribute to both innate and adaptive immune responses. With mediated expression, it can also control inflammation. It is potent governing resistance in early developmental days as well. Its expression in pituitary and testis suggests its participation in reproduction and endocrine regulation in fish. Overall, β-defensins is an important member of antimicrobial peptides (AMPs) with multifunctional role in general homeostasis and to pathogen exposure possessing tremendous therapeutic approaches.
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Affiliation(s)
- Sweta Das
- Department of Aquatic Animal Health & Management, Kerala University of Fisheries and Ocean Studies, Kochi, Kerala, India.
| | - Chiranjiv Pradhan
- Department of Aquaculture, Kerala University of Fisheries and Ocean Studies, Kochi, Kerala, India
| | - Devika Pillai
- Department of Aquatic Animal Health & Management, Kerala University of Fisheries and Ocean Studies, Kochi, Kerala, India
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11
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Laghi V, Rezelj V, Boucontet L, Frétaud M, Da Costa B, Boudinot P, Salinas I, Lutfalla G, Vignuzzi M, Levraud JP. Exploring Zebrafish Larvae as a COVID-19 Model: Probable Abortive SARS-CoV-2 Replication in the Swim Bladder. Front Cell Infect Microbiol 2022; 12:790851. [PMID: 35360100 PMCID: PMC8963489 DOI: 10.3389/fcimb.2022.790851] [Citation(s) in RCA: 9] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/07/2021] [Accepted: 02/04/2022] [Indexed: 12/16/2022] Open
Abstract
Animal models are essential to understanding COVID-19 pathophysiology and for preclinical assessment of drugs and other therapeutic or prophylactic interventions. We explored the small, cheap, and transparent zebrafish larva as a potential host for SARS-CoV-2. Bath exposure, as well as microinjection in the coelom, pericardium, brain ventricle, or bloodstream, resulted in a rapid decrease of SARS-CoV-2 RNA in wild-type larvae. However, when the virus was inoculated in the swim bladder, viral RNA stabilized after 24 h. By immunohistochemistry, epithelial cells containing SARS-CoV-2 nucleoprotein were observed in the swim bladder wall. Our data suggest an abortive infection of the swim bladder. In some animals, several variants of concern were also tested with no evidence of increased infectivity in our model. Low infectivity of SARS-CoV-2 in zebrafish larvae was not due to the host type I interferon response, as comparable viral loads were detected in type I interferon-deficient animals. A mosaic overexpression of human ACE2 was not sufficient to increase SARS-CoV-2 infectivity in zebrafish embryos or in fish cells in vitro. In conclusion, wild-type zebrafish larvae appear mostly non-permissive to SARS-CoV-2, except in the swim bladder, an aerial organ sharing similarities with the mammalian lung.
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Affiliation(s)
- Valerio Laghi
- Institut Pasteur, Centre National de la Recherche Scientifique (CNRS) Unité mixte de Recherche (UMR) 3637, Unité Macrophages et Développement de l’Immunité, Paris, France
| | - Veronica Rezelj
- Institut Pasteur, Unité Populations Virales et Pathogénèse, Institut Pasteur, Paris, France
| | - Laurent Boucontet
- Institut Pasteur, Centre National de la Recherche Scientifique (CNRS) Unité mixte de Recherche (UMR) 3637, Unité Macrophages et Développement de l’Immunité, Paris, France
| | - Maxence Frétaud
- Université Paris-Saclay, Institut National pour la Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université Versailles Saint-Quentin (UVSQ), Virologie et Immunologie Moléculaire (VIM), Jouy-en-Josas, France
| | - Bruno Da Costa
- Université Paris-Saclay, Institut National pour la Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université Versailles Saint-Quentin (UVSQ), Virologie et Immunologie Moléculaire (VIM), Jouy-en-Josas, France
| | - Pierre Boudinot
- Université Paris-Saclay, Institut National pour la Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université Versailles Saint-Quentin (UVSQ), Virologie et Immunologie Moléculaire (VIM), Jouy-en-Josas, France
| | - Irene Salinas
- Department of Biology, University of New Mexico, Albuquerque, NM, United States
| | - Georges Lutfalla
- Laboratory of Pathogen-Host Interactions (LPHI), Centre National de la Recherche Scientifique (CNRS), Université de Montpellier, Montpellier, France
| | - Marco Vignuzzi
- Institut Pasteur, Unité Populations Virales et Pathogénèse, Institut Pasteur, Paris, France
| | - Jean-Pierre Levraud
- Institut Pasteur, Centre National de la Recherche Scientifique (CNRS) Unité mixte de Recherche (UMR) 3637, Unité Macrophages et Développement de l’Immunité, Paris, France
- Université Paris-Saclay, Centre National de la Recherche Scientifique (CNRS), Institut Pasteur, Institut des Neurosciences Paris-Saclay, Gif-sur-Yvette, France
- *Correspondence: Jean-Pierre Levraud,
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12
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Laghi V, Rezelj V, Boucontet L, Frétaud M, Da Costa B, Boudinot P, Salinas I, Lutfalla G, Vignuzzi M, Levraud JP. Exploring Zebrafish Larvae as a COVID-19 Model: Probable Abortive SARS-CoV-2 Replication in the Swim Bladder. Front Cell Infect Microbiol 2022. [PMID: 35360100 DOI: 10.1101/2021.04.08.439059v1] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 05/07/2023] Open
Abstract
Animal models are essential to understanding COVID-19 pathophysiology and for preclinical assessment of drugs and other therapeutic or prophylactic interventions. We explored the small, cheap, and transparent zebrafish larva as a potential host for SARS-CoV-2. Bath exposure, as well as microinjection in the coelom, pericardium, brain ventricle, or bloodstream, resulted in a rapid decrease of SARS-CoV-2 RNA in wild-type larvae. However, when the virus was inoculated in the swim bladder, viral RNA stabilized after 24 h. By immunohistochemistry, epithelial cells containing SARS-CoV-2 nucleoprotein were observed in the swim bladder wall. Our data suggest an abortive infection of the swim bladder. In some animals, several variants of concern were also tested with no evidence of increased infectivity in our model. Low infectivity of SARS-CoV-2 in zebrafish larvae was not due to the host type I interferon response, as comparable viral loads were detected in type I interferon-deficient animals. A mosaic overexpression of human ACE2 was not sufficient to increase SARS-CoV-2 infectivity in zebrafish embryos or in fish cells in vitro. In conclusion, wild-type zebrafish larvae appear mostly non-permissive to SARS-CoV-2, except in the swim bladder, an aerial organ sharing similarities with the mammalian lung.
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Affiliation(s)
- Valerio Laghi
- Institut Pasteur, Centre National de la Recherche Scientifique (CNRS) Unité mixte de Recherche (UMR) 3637, Unité Macrophages et Développement de l'Immunité, Paris, France
| | - Veronica Rezelj
- Institut Pasteur, Unité Populations Virales et Pathogénèse, Institut Pasteur, Paris, France
| | - Laurent Boucontet
- Institut Pasteur, Centre National de la Recherche Scientifique (CNRS) Unité mixte de Recherche (UMR) 3637, Unité Macrophages et Développement de l'Immunité, Paris, France
| | - Maxence Frétaud
- Université Paris-Saclay, Institut National pour la Recherche pour l'Agriculture, l'Alimentation et l'Environnement (INRAE), Université Versailles Saint-Quentin (UVSQ), Virologie et Immunologie Moléculaire (VIM), Jouy-en-Josas, France
| | - Bruno Da Costa
- Université Paris-Saclay, Institut National pour la Recherche pour l'Agriculture, l'Alimentation et l'Environnement (INRAE), Université Versailles Saint-Quentin (UVSQ), Virologie et Immunologie Moléculaire (VIM), Jouy-en-Josas, France
| | - Pierre Boudinot
- Université Paris-Saclay, Institut National pour la Recherche pour l'Agriculture, l'Alimentation et l'Environnement (INRAE), Université Versailles Saint-Quentin (UVSQ), Virologie et Immunologie Moléculaire (VIM), Jouy-en-Josas, France
| | - Irene Salinas
- Department of Biology, University of New Mexico, Albuquerque, NM, United States
| | - Georges Lutfalla
- Laboratory of Pathogen-Host Interactions (LPHI), Centre National de la Recherche Scientifique (CNRS), Université de Montpellier, Montpellier, France
| | - Marco Vignuzzi
- Institut Pasteur, Unité Populations Virales et Pathogénèse, Institut Pasteur, Paris, France
| | - Jean-Pierre Levraud
- Institut Pasteur, Centre National de la Recherche Scientifique (CNRS) Unité mixte de Recherche (UMR) 3637, Unité Macrophages et Développement de l'Immunité, Paris, France
- Université Paris-Saclay, Centre National de la Recherche Scientifique (CNRS), Institut Pasteur, Institut des Neurosciences Paris-Saclay, Gif-sur-Yvette, France
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13
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Okamura Y, Miyanishi H, Kono T, Sakai M, Hikima JI. Identification and expression of phospholipase A2 genes related to transcriptional control in the interleukin-17A/F1 pathway in the intestines of Japanese medaka Oryzias latipes. FISH AND SHELLFISH IMMUNOLOGY REPORTS 2021; 2:100028. [PMID: 36420487 PMCID: PMC9680080 DOI: 10.1016/j.fsirep.2021.100028] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/10/2021] [Revised: 09/09/2021] [Accepted: 09/09/2021] [Indexed: 12/16/2022] Open
Abstract
Phospholipase A2 (PLA2), a phospholipid hydrolase, has recently attracted attention owing to its broad functionality. Immunological evidence has revealed increased susceptibility to infectious diseases and immunodeficiency in knockout (KO) mice of several pla2 genes. However, no progress has been made in terms of immunological research on any pla2 gene in fish. In this study, we focused on the intestinal immune responses of fish PLA2s. The full-length open reading frames of pla2g1b, pla2g3, pla2g10, pla2g12b1, pla2g12b2, and pla2g15 cDNAs were cloned in Japanese medaka (Orizias latipes), and their gene expressions were quantified by real-time PCR (qPCR) and in situ hybridization (ISH). Characterization of pla2 genes revealed a functional domain and three-dimensional structure similar to the mammalian counterparts. In addition, expression of pla2g1b, pla2g12b1, and pla2g12b2 was extremely high in Japanese medaka intestines. ISH detected strong expression of pla2g1b mRNAs in the basal muscle layer, and pla2g12b1 and pla2g12b2 mRNAs were detected in the epithelial cells. In the medaka exposed to Edwardsiella piscicida, pla2g12b1, pla2g12b2 and pla2g15 were significantly induced in the anterior and posterior intestines, and pla2g1b was upregulated in the anterior intestine. Furthermore, pla2g1b, pla2g3, pla2g10, and pla2g12b2 were significantly downregulated in the IL-17A/F1 KO medaka compared to those in wild-type medaka. These results suggest that these PLA2s are involved in intestinal immunity in teleosts.
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Affiliation(s)
- Yo Okamura
- Interdisciplinary Graduate School of Agriculture and Engineering, University of Miyazaki, Miyazaki, Japan
| | - Hiroshi Miyanishi
- Department of Marine Biology and Environmental Science, Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan
| | - Tomoya Kono
- Department of Biochemistry and Applied Biosciences, Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan
| | - Masahiro Sakai
- Department of Biochemistry and Applied Biosciences, Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan
| | - Jun-ichi Hikima
- Department of Biochemistry and Applied Biosciences, Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan
- Corresponding author.
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14
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Graves CL, Chen A, Kwon V, Shiau CE. Zebrafish harbor diverse intestinal macrophage populations including a subset intimately associated with enteric neural processes. iScience 2021; 24:102496. [PMID: 34142024 PMCID: PMC8185245 DOI: 10.1016/j.isci.2021.102496] [Citation(s) in RCA: 11] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/20/2020] [Revised: 03/17/2021] [Accepted: 04/28/2021] [Indexed: 02/07/2023] Open
Abstract
Intestinal macrophages are essential for gut health but remain understudied outside of human and mouse systems. Here, we establish zebrafish as a powerful model that provides superior imaging capabilities for whole-gut analysis along all dimensions (anterior-posterior and center-outer axes) for dissecting macrophage biology in gastrointestinal health and disease. We utilized high-resolution imaging to show that the zebrafish gut contains bona fide muscularis and mucosal macrophages, as well as surprisingly large subsets intimately associated with enteric neural processes. Interestingly, most muscularis macrophages span multiple gut layers in stark contrast to their mammalian counterparts typically restricted to a single layer. Using macrophage-deficient irf8 zebrafish, we found a depletion of muscularis but not mucosal macrophages, and that they may be dispensable for gross intestinal transit in adults but not during development. These characterizations provide first insights into intestinal macrophages and their association with the enteric nervous system from development to adulthood in teleosts.
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Affiliation(s)
- Christina L. Graves
- Department of Biology, University of North Carolina Chapel Hill, Chapel Hill, NC 27599, USA
| | - Angela Chen
- Department of Biology, University of North Carolina Chapel Hill, Chapel Hill, NC 27599, USA
| | - Victoria Kwon
- Department of Biology, University of North Carolina Chapel Hill, Chapel Hill, NC 27599, USA
| | - Celia E. Shiau
- Department of Biology, University of North Carolina Chapel Hill, Chapel Hill, NC 27599, USA
- Department of Microbiology and Immunology, University of North Carolina Chapel Hill, Chapel Hill, NC 27599, USA
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15
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Okamura Y, Miyanishi H, Kinoshita M, Kono T, Sakai M, Hikima JI. A defective interleukin-17 receptor A1 causes weight loss and intestinal metabolism-related gene downregulation in Japanese medaka, Oryzias latipes. Sci Rep 2021; 11:12099. [PMID: 34103614 PMCID: PMC8187396 DOI: 10.1038/s41598-021-91534-3] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/23/2020] [Accepted: 05/27/2021] [Indexed: 02/08/2023] Open
Abstract
In the intestine, the host must be able to control the gut microbiota and efficiently absorb transiently supplied metabolites, at the risk of enormous infection. In mammals, the inflammatory cytokine interleukin (IL)-17A/F is one of the key mediators in the intestinal immune system. However, many functions of IL-17 in vertebrate intestines remain unclarified. In this study, we established a gene-knockout (KO) model of IL-17 receptor A1 (IL-17RA1, an IL-17A/F receptor) in Japanese medaka (Oryzias latipes) using genome editing technique, and the phenotypes were compared to wild type (WT) based on transcriptome analyses. Upon hatching, homozygous IL-17RA1-KO medaka mutants showed no significant morphological abnormality. However, after 4 months, significant weight decreases and reduced survival rates were observed in IL-17RA1-KO medaka. Comparison of gene-expression patterns in WT and IL-17RA1-KO medaka revealed that various metabolism- and immune-related genes were significantly down-regulated in IL-17RA1-KO medaka intestine, particularly genes related to mevalonate metabolism (mvda, acat2, hmgcs1, and hmgcra) and genes related to IL-17 signaling (such as il17c, il17a/f1, and rorc) were found to be decreased. Conversely, expression of genes related to cardiovascular system development, including fli1a, sox7, and notch1b in the anterior intestine, and that of genes related to oxidation-reduction processes including ugp2a, aoc1, and nos1 in posterior intestine was up-regulated in IL-17RA1-KO medaka. These findings show that IL-17RA regulated immune- and various metabolism-related genes in the intestine for maintaining the health of Japanese medaka.
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Affiliation(s)
- Yo Okamura
- Interdisciplinary Graduate School of Agriculture and Engineering, University of Miyazaki, Miyazaki, Japan
| | - Hiroshi Miyanishi
- Department of Marine Biology and Environmental Science, Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan
| | - Masato Kinoshita
- Division of Applied Biosciences, Graduate School of Agriculture , Kyoto University, Kyoto, Japan
| | - Tomoya Kono
- Department of Biochemistry and Applied Biosciences, Faculty of Agriculture , University of Miyazaki, Miyazaki, Japan
| | - Masahiro Sakai
- Department of Biochemistry and Applied Biosciences, Faculty of Agriculture , University of Miyazaki, Miyazaki, Japan
| | - Jun-Ichi Hikima
- Department of Biochemistry and Applied Biosciences, Faculty of Agriculture , University of Miyazaki, Miyazaki, Japan.
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16
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Gu H, Feng Y, Zhang Y, Yin D, Yang Z, Tang W. Differential study of the Parabramis pekinensis intestinal microbiota according to different habitats and different parts of the intestine. ANN MICROBIOL 2021. [DOI: 10.1186/s13213-020-01614-4] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/11/2022] Open
Abstract
Abstract
Purpose
To identify the differences in gut bacterial community of Parabramis pekinensis under different growth conditions, and the effect of the diet in a controlled habitat on the community structure, aiming to provide a comprehensive survey of how the gut microbiota in P. pekinensis varies depending on habitat.
Methods
A total of 73 P. pekinensis from Yangtze River (W), rivers in the outskirts of Jingjiang (Jiangsu province, China, R), and farms (C) were collected to analyze the intestinal microbiota using high-throughput sequencing of the V3-V4 16S ribosomal RNA gene. We also subdivided the gut into the foregut (F), midgut (M), and hindgut (B) to analyze the differences between them.
Results
The dominant bacterial phyla in P. pekinensis were Fusobacteria, Firmicutes, Proteobacteria, and Actinobacteria; meanwhile, Cyanobacteria, Bacteroidetes, Chloroflexi, and Verrucomicrobia were also highly abundant. It is worth noting that the abundance of Fusobacteria Cetobacterium was also very high. The abundance and diversity of the intestinal microbiota structure of fish taken from breeding farm were significantly lower than those taken from Yangtze river and Suburban river, and the abundance of Aeromonas in the gut of fish taken from Yangtze river was much higher than that of fish taken from Suburban river. Compared to midgut, foregut and hindgut have similar microbiota structures, but did not differ significantly in them.
Conclusions
The core intestinal microbiota of P. pekinensis is the same to other herbivorous and partially omnivorous fish. There were significant differences in the intestinal microbiota structure of P. pekinensis from different habitats, but no significant differences in the microbiota abundance and diversity between the different parts of the intestine.
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17
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Picchietti S, Miccoli A, Fausto AM. Gut immunity in European sea bass (Dicentrarchus labrax): a review. FISH & SHELLFISH IMMUNOLOGY 2021; 108:94-108. [PMID: 33285171 DOI: 10.1016/j.fsi.2020.12.001] [Citation(s) in RCA: 14] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 07/23/2020] [Revised: 12/01/2020] [Accepted: 12/02/2020] [Indexed: 06/12/2023]
Abstract
In this review, we summarize and discuss the trends and supporting findings in scientific literature on the gut mucosa immune role in European sea bass (Dicentrarchus labrax L.). Overall, the purpose is to provide an updated overview of the gastrointestinal tract functional regionalization and defence barriers. A description of the available information regarding immune cells found in two immunologically-relevant intestinal compartments, namely epithelium and lamina propria, is provided. Attention has been also paid to mucosal immunoglobulins and to the latest research investigating gut microbiota and dietary manipulation impacts. Finally, we review oral vaccination strategies, as a safe method for sea bass vaccine delivery.
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Affiliation(s)
- S Picchietti
- Department for Innovation in Biological, Agro-food and Forest Systems (DIBAF), University of Tuscia, Viterbo, Italy.
| | - A Miccoli
- Department for Innovation in Biological, Agro-food and Forest Systems (DIBAF), University of Tuscia, Viterbo, Italy
| | - A M Fausto
- Department for Innovation in Biological, Agro-food and Forest Systems (DIBAF), University of Tuscia, Viterbo, Italy
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18
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Cholan PM, Han A, Woodie BR, Watchon M, Kurz AR, Laird AS, Britton WJ, Ye L, Holmes ZC, McCann JR, David LA, Rawls JF, Oehlers SH. Conserved anti-inflammatory effects and sensing of butyrate in zebrafish. Gut Microbes 2020; 12:1-11. [PMID: 33064972 PMCID: PMC7575005 DOI: 10.1080/19490976.2020.1824563] [Citation(s) in RCA: 50] [Impact Index Per Article: 10.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/03/2023] Open
Abstract
Short-chain fatty acids (SCFAs) are produced by microbial fermentation of dietary fiber in the gut. Butyrate is a particularly important SCFA with anti-inflammatory properties and is generally present at lower levels in inflammatory diseases associated with gut microbiota dysbiosis in mammals. We aimed to determine if SCFAs are produced by the zebrafish microbiome and if SCFAs exert conserved effects on zebrafish immunity as an example of the non-mammalian vertebrate immune system. We demonstrate that bacterial communities from adult zebrafish intestines synthesize all three main SCFA in vitro, although SCFA were below our detectable limits in zebrafish intestines in vivo. Immersion in butyrate, but not acetate or propionate, reduced the recruitment of neutrophils and M1-type pro-inflammatory macrophages to wounds. We found conservation of butyrate sensing by neutrophils via orthologs of the hydroxycarboxylic acid receptor 1 (hcar1) gene. Neutrophils from Hcar1-depleted embryos were no longer responsive to the anti-inflammatory effects of butyrate, while macrophage sensitivity to butyrate was independent of Hcar1. Our data demonstrate conservation of anti-inflammatory butyrate effects and identify the presence of a conserved molecular receptor in fish.
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Affiliation(s)
- Pradeep Manuneedhi Cholan
- Tuberculosis Research Program at the Centenary Institute, The University of Sydney , Camperdown, Australia
| | - Alvin Han
- Department of Molecular Genetics and Microbiology, Duke Microbiome Center, Duke University School of Medicine , Durham, NC, USA
| | - Brad R Woodie
- Tuberculosis Research Program at the Centenary Institute, The University of Sydney , Camperdown, Australia.,Department of Molecular Genetics and Microbiology, Duke Microbiome Center, Duke University School of Medicine , Durham, NC, USA
| | - Maxinne Watchon
- Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University , Macquarie Park, Australia.,Sydney Medical School, The University of Sydney , Camperdown, Australia
| | - Angela Rm Kurz
- Centenary Imaging and Sydney Cytometry at the Centenary Institute, The University of Sydney , Camperdown, Australia
| | - Angela S Laird
- Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University , Macquarie Park, Australia
| | - Warwick J Britton
- Tuberculosis Research Program at the Centenary Institute, The University of Sydney , Camperdown, Australia.,The University of Sydney, Discipline of Infectious Diseases and Immunology, Faculty of Medicine and Health, And Marie Bashir Institute , Camperdown, Australia.,Department of Clinical Immunology, Royal Prince Alfred Hospital , Camperdown, Australia
| | - Lihua Ye
- Department of Molecular Genetics and Microbiology, Duke Microbiome Center, Duke University School of Medicine , Durham, NC, USA
| | - Zachary C Holmes
- Department of Molecular Genetics and Microbiology, Duke Microbiome Center, Duke University School of Medicine , Durham, NC, USA
| | - Jessica R McCann
- Department of Molecular Genetics and Microbiology, Duke Microbiome Center, Duke University School of Medicine , Durham, NC, USA
| | - Lawrence A David
- Department of Molecular Genetics and Microbiology, Duke Microbiome Center, Duke University School of Medicine , Durham, NC, USA
| | - John F Rawls
- Department of Molecular Genetics and Microbiology, Duke Microbiome Center, Duke University School of Medicine , Durham, NC, USA
| | - Stefan H Oehlers
- Tuberculosis Research Program at the Centenary Institute, The University of Sydney , Camperdown, Australia.,The University of Sydney, Discipline of Infectious Diseases and Immunology, Faculty of Medicine and Health, And Marie Bashir Institute , Camperdown, Australia
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19
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Kamareddine L, Najjar H, Sohail MU, Abdulkader H, Al-Asmakh M. The Microbiota and Gut-Related Disorders: Insights from Animal Models. Cells 2020; 9:cells9112401. [PMID: 33147801 PMCID: PMC7693214 DOI: 10.3390/cells9112401] [Citation(s) in RCA: 24] [Impact Index Per Article: 4.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/03/2020] [Revised: 10/23/2020] [Accepted: 10/29/2020] [Indexed: 02/06/2023] Open
Abstract
Over the past decade, the scientific committee has called for broadening our horizons in understanding host–microbe interactions and infectious disease progression. Owing to the fact that the human gut harbors trillions of microbes that exhibit various roles including the production of vitamins, absorption of nutrients, pathogen displacement, and development of the host immune system, particular attention has been given to the use of germ-free (GF) animal models in unraveling the effect of the gut microbiota on the physiology and pathophysiology of the host. In this review, we discuss common methods used to generate GF fruit fly, zebrafish, and mice model systems and highlight the use of these GF model organisms in addressing the role of gut-microbiota in gut-related disorders (metabolic diseases, inflammatory bowel disease, and cancer), and in activating host defense mechanisms and amending pathogenic virulence.
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Affiliation(s)
- Layla Kamareddine
- Department of Biomedical Science, College of Health Sciences, QU Health, Qatar University, P.O. Box 2713 Doha, Qatar; (L.K.); (H.N.); (M.U.S.); (H.A.)
| | - Hoda Najjar
- Department of Biomedical Science, College of Health Sciences, QU Health, Qatar University, P.O. Box 2713 Doha, Qatar; (L.K.); (H.N.); (M.U.S.); (H.A.)
| | - Muhammad Umar Sohail
- Department of Biomedical Science, College of Health Sciences, QU Health, Qatar University, P.O. Box 2713 Doha, Qatar; (L.K.); (H.N.); (M.U.S.); (H.A.)
- Biomedical Research Center, QU Health, Qatar University, P.O. Box 2713 Doha, Qatar
| | - Hadil Abdulkader
- Department of Biomedical Science, College of Health Sciences, QU Health, Qatar University, P.O. Box 2713 Doha, Qatar; (L.K.); (H.N.); (M.U.S.); (H.A.)
| | - Maha Al-Asmakh
- Department of Biomedical Science, College of Health Sciences, QU Health, Qatar University, P.O. Box 2713 Doha, Qatar; (L.K.); (H.N.); (M.U.S.); (H.A.)
- Biomedical Research Center, QU Health, Qatar University, P.O. Box 2713 Doha, Qatar
- Correspondence: ; Tel.: +974-4403-4789
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20
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Okamura Y, Morimoto N, Sawada S, Kono T, Hikima JI, Sakai M. Molecular characterization and expression of two interleukin-17 receptor A genes on different chromosomes in Japanese medaka, Oryzias latipes. Comp Biochem Physiol B Biochem Mol Biol 2020; 240:110386. [DOI: 10.1016/j.cbpb.2019.110386] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/13/2019] [Revised: 11/03/2019] [Accepted: 11/08/2019] [Indexed: 12/18/2022]
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21
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Flores EM, Nguyen AT, Odem MA, Eisenhoffer GT, Krachler AM. The zebrafish as a model for gastrointestinal tract-microbe interactions. Cell Microbiol 2020; 22:e13152. [PMID: 31872937 DOI: 10.1111/cmi.13152] [Citation(s) in RCA: 68] [Impact Index Per Article: 13.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/30/2019] [Revised: 11/07/2019] [Accepted: 12/19/2019] [Indexed: 02/06/2023]
Abstract
The zebrafish (Danio rerio) has become a widely used vertebrate model for bacterial, fungal, viral, and protozoan infections. Due to its genetic tractability, large clutch sizes, ease of manipulation, and optical transparency during early life stages, it is a particularly useful model to address questions about the cellular microbiology of host-microbe interactions. Although its use as a model for systemic infections, as well as infections localised to the hindbrain and swimbladder having been thoroughly reviewed, studies focusing on host-microbe interactions in the zebrafish gastrointestinal tract have been neglected. Here, we summarise recent findings regarding the developmental and immune biology of the gastrointestinal tract, drawing parallels to mammalian systems. We discuss the use of adult and larval zebrafish as models for gastrointestinal infections, and more generally, for studies of host-microbe interactions in the gut.
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Affiliation(s)
- Erika M Flores
- Department of Microbiology and Molecular Genetics, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, Texas.,M.D. Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences, Houston, Texas
| | - Anh T Nguyen
- Department of Microbiology and Molecular Genetics, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, Texas.,M.D. Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences, Houston, Texas
| | - Max A Odem
- Department of Microbiology and Molecular Genetics, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, Texas
| | - George T Eisenhoffer
- M.D. Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences, Houston, Texas.,Department of Genetics, University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Anne Marie Krachler
- Department of Microbiology and Molecular Genetics, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, Texas.,M.D. Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences, Houston, Texas
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22
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Espenschied ST, Cronan MR, Matty MA, Mueller O, Redinbo MR, Tobin DM, Rawls JF. Epithelial delamination is protective during pharmaceutical-induced enteropathy. Proc Natl Acad Sci U S A 2019; 116:16961-16970. [PMID: 31391308 PMCID: PMC6708343 DOI: 10.1073/pnas.1902596116] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022] Open
Abstract
Intestinal epithelial cell (IEC) shedding is a fundamental response to intestinal damage, yet underlying mechanisms and functions have been difficult to define. Here we model chronic intestinal damage in zebrafish larvae using the nonsteroidal antiinflammatory drug (NSAID) Glafenine. Glafenine induced the unfolded protein response (UPR) and inflammatory pathways in IECs, leading to delamination. Glafenine-induced inflammation was augmented by microbial colonization and associated with changes in intestinal and environmental microbiotas. IEC shedding was a UPR-dependent protective response to Glafenine that restricts inflammation and promotes animal survival. Other NSAIDs did not induce IEC delamination; however, Glafenine also displays off-target inhibition of multidrug resistance (MDR) efflux pumps. We found a subset of MDR inhibitors also induced IEC delamination, implicating MDR efflux pumps as cellular targets underlying Glafenine-induced enteropathy. These results implicate IEC delamination as a protective UPR-mediated response to chemical injury, and uncover an essential role for MDR efflux pumps in intestinal homeostasis.
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Affiliation(s)
- Scott T Espenschied
- Department of Molecular Genetics and Microbiology, Duke University School of Medicine, Durham, NC 27710
| | - Mark R Cronan
- Department of Molecular Genetics and Microbiology, Duke University School of Medicine, Durham, NC 27710
| | - Molly A Matty
- Department of Molecular Genetics and Microbiology, Duke University School of Medicine, Durham, NC 27710
| | - Olaf Mueller
- Department of Molecular Genetics and Microbiology, Duke University School of Medicine, Durham, NC 27710
| | - Matthew R Redinbo
- Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599
- Department of Biochemistry, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC 27599
- Department of Microbiology and Immunology, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC 27599
| | - David M Tobin
- Department of Molecular Genetics and Microbiology, Duke University School of Medicine, Durham, NC 27710
- Department of Medicine, Duke University School of Medicine, Durham, NC 27710
- Department of Immunology, Duke University School of Medicine, Durham, NC 27710
| | - John F Rawls
- Department of Molecular Genetics and Microbiology, Duke University School of Medicine, Durham, NC 27710;
- Department of Medicine, Duke University School of Medicine, Durham, NC 27710
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23
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Intravital Imaging Reveals Divergent Cytokine and Cellular Immune Responses to Candida albicans and Candida parapsilosis. mBio 2019; 10:mBio.00266-19. [PMID: 31088918 PMCID: PMC6520444 DOI: 10.1128/mbio.00266-19] [Citation(s) in RCA: 15] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
In modern medicine, physicians are frequently forced to balance immune suppression against immune stimulation to treat patients such as those undergoing transplants and chemotherapy. More-targeted therapies designed to preserve immunity and prevent opportunistic fungal infection in these patients could be informed by an understanding of how fungi interact with professional and nonprofessional immune cells in mucosal candidiasis. In this study, we intravitally imaged these host-pathogen dynamics during Candida infection in a transparent vertebrate model host, the zebrafish. Single-cell imaging revealed an unexpected partitioning of the inflammatory response between phagocytes and epithelial cells. Surprisingly, we found that in vivo cytokine profiles more closely match in vitro responses of epithelial cells rather than phagocytes. Furthermore, we identified a disconnect between canonical inflammatory cytokine production and phagocyte recruitment to the site of infection, implicating noncytokine chemoattractants. Our study contributes to a new appreciation for the specialization and cross talk among cell types during mucosal infection. Candida yeasts are common commensals that can cause mucosal disease and life-threatening systemic infections. While many of the components required for defense against Candida albicans infection are well established, questions remain about how various host cells at mucosal sites assess threats and coordinate defenses to prevent normally commensal organisms from becoming pathogenic. Using two Candida species, C. albicans and C. parapsilosis, which differ in their abilities to damage epithelial tissues, we used traditional methods (pathogen CFU, host survival, and host cytokine expression) combined with high-resolution intravital imaging of transparent zebrafish larvae to illuminate host-pathogen interactions at the cellular level in the complex environment of a mucosal infection. In zebrafish, C. albicans grows as both yeast and epithelium-damaging filaments, activates the NF-κB pathway, evokes proinflammatory cytokines, and causes the recruitment of phagocytic immune cells. On the other hand, C. parapsilosis remains in yeast morphology and elicits the recruitment of phagocytes without inducing inflammation. High-resolution mapping of phagocyte-Candida interactions at the infection site revealed that neutrophils and macrophages attack both Candida species, regardless of the cytokine environment. Time-lapse monitoring of single-cell gene expression in transgenic reporter zebrafish revealed a partitioning of the immune response during C. albicans infection: the transcription factor NF-κB is activated largely in cells of the swimbladder epithelium, while the proinflammatory cytokine tumor necrosis factor alpha (TNF-α) is expressed in motile cells, mainly macrophages. Our results point to different host strategies for combatting pathogenic Candida species and separate signaling roles for host cell types.
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24
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Mohammadian T, Jangaran-Nejad A, Mesbah M, Shirali T, Malekpouri P, Tabandeh MR. Effect of Lactobacillus casei on Innate Immunity Responses and Aeromonas hydrophila Resistance in Shabot, Tor grypus. Probiotics Antimicrob Proteins 2019; 12:224-235. [PMID: 30637563 DOI: 10.1007/s12602-018-9510-z] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/08/2023]
Abstract
For investigating the possible effect of Lactobacillus casei on the immune status of Shabot, Tor grypus, 480 fish weighing 40 ± 10 g were randomly divided into four groups in triplicate. The first group (control) was fed on a commercial diet without probiotic supplementation. Other groups (A, B, and C) were received diet supplemented by different levels of L. casei, i.e., 5 × 106, 5 × 107, and 5 × 108 CFU g-1, respectively. The feeding trial was performed for 60 continuous days followed by 15 days as withdraw period, i.e., day 75. Serum immune responses were determined on days 0, 30, 60, and 75 of feeding trial. Gene expression in the kidneys was analyzed on days 0, 60, and 75 as well as following bacterial challenge. The results showed that dietary L. casei supplementation significantly (P < 0.05) increased serum lysozyme, serum bactericidal, complement, and respiratory burst activities as compared with control group, while it was observed that treated fish did not show any significant differences in immune responses on day 75 (P < 0.05). Results of gene expression (TNF-α, IL-1β, and IL-8) showed significant increases among different experimental times (P < 0.05). According to obtained results, it might be concluded that the feeding by L. casei could likely enhance the immune responses and gene expression in T. grypus. Treatments A and C were better than B, but the lower concentration is better as an economic viewpoints and biosecurity. Therefore, concentration A is suggested.
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Affiliation(s)
- Takavar Mohammadian
- Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, 61357-831351, Iran.
| | - Abdolhossein Jangaran-Nejad
- Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, 61357-831351, Iran
| | - Mehrzad Mesbah
- Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, 61357-831351, Iran
| | - Tahereh Shirali
- Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, 61357-831351, Iran
| | - Pedram Malekpouri
- Young Researchers and Elites Club, Science and Research Branch, Islamic Azad University, Tehran, Iran
| | - Mohammad-Reza Tabandeh
- Department of Biochemistry and Molecular Biology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran
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25
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Mowbray CA, Niranji SS, Cadwell K, Bailey R, Watson KA, Hall J. Gene expression of AvBD6-10 in broiler chickens is independent of AvBD6, 9, and 10 peptide potency. Vet Immunol Immunopathol 2018; 202:31-40. [PMID: 30078596 DOI: 10.1016/j.vetimm.2018.06.007] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/09/2018] [Revised: 05/23/2018] [Accepted: 06/07/2018] [Indexed: 02/07/2023]
Abstract
The Avian β-defensin (AvBD) gene cluster contains fourteen genes; within this, two groups (AvBD6/7 and AvBD8 -10) encode charged peptides of >+5 (AvBD6/7), indicative of potent microbial killing activities, and ≤+4 (AvBD8-10), suggestive of reduced antimicrobial activities. Chicken broiler gut tissues are constantly exposed to microbes in the form of commensal bacteria. This study examined whether tissue expression patterns of AvBD6-10 reflected microbial exposure and the encoded peptides a functional antimicrobial hierarchy. Gut AvBD6-10 gene expression was observed in hatch to day 21 birds, although the AvBD8-10 profiles were eclipsed by those detected in the liver and kidney tissues. In vitro challenges of chicken CHCC-OU2 cells using the gut commensal Lactobacillus johnsonii (104 CFU) did not significantly affect AvBD8-10 gene expression patterns, although upregulation (P < 0.05) of IL-Iβ gene expression was observed. Similarly, in response to Bacteriodes doreii, IL-Iβ and IL-6 gene upregulation were detected (P < 0.05), but AvBD10 gene expression remained unaffected. These data suggested that AvBD8-10 gene expression was not induced by commensal gut bacteria. Bacterial time-kill assays employing recombinant (r)AvBD6, 9 and 10 peptides (0.5μM - 12μM), indicated an antimicrobial hierarchy, linked to charge, of AvBD6 > AvBD9 > AvBD10 against Escherichia coli, but AvBD10 > AvBD9 > AvBD6 using Enterococcus faecalis. rAvBD10, selected due to its reduced cationic charge was, using CHCC-OU2 cells, investigated for cell proliferation and wound healing properties, but none were observed. These data suggest that in healthy broiler chicken tissues AvBD6/7 and AvBD8-10 gene expression profiles are independent of the in vitro antimicrobial hierarchies of the encoded AvBD6, 9 and 10 peptides.
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Affiliation(s)
- Catherine A Mowbray
- Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle Upon Tyne, NE2 4HH, UK
| | - Sherko S Niranji
- Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle Upon Tyne, NE2 4HH, UK
| | - Kevin Cadwell
- Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle Upon Tyne, NE2 4HH, UK
| | | | | | - Judith Hall
- Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle Upon Tyne, NE2 4HH, UK.
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26
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de Abreu MS, Giacomini ACVV, Zanandrea R, Dos Santos BE, Genario R, de Oliveira GG, Friend AJ, Amstislavskaya TG, Kalueff AV. Psychoneuroimmunology and immunopsychiatry of zebrafish. Psychoneuroendocrinology 2018; 92:1-12. [PMID: 29609110 DOI: 10.1016/j.psyneuen.2018.03.014] [Citation(s) in RCA: 15] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/23/2018] [Revised: 03/14/2018] [Accepted: 03/21/2018] [Indexed: 12/11/2022]
Abstract
Despite the high prevalence of neural and immune disorders, their etiology and molecular mechanisms remain poorly understood. As the zebrafish (Danio rerio) is increasingly utilized as a powerful model organism in biomedical research, mounting evidence suggests these fish as a useful tool to study neural and immune mechanisms and their interplay. Here, we discuss zebrafish neuro-immune mechanisms and their pharmacological and genetic modulation, the effect of stress on cytokines, as well as relevant models of microbiota-brain interplay. As many human brain diseases are based on complex interplay between the neural and the immune system, here we discuss zebrafish models, as well as recent successes and challenges, in this rapidly expanding field. We particularly emphasize the growing utility of zebrafish models in translational immunopsychiatry research, as they improve our understanding of pathogenetic neuro-immune interactions, thereby fostering future discovery of potential therapeutic agents.
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Affiliation(s)
- Murilo S de Abreu
- Bioscience Institute, University of Passo Fundo (UPF), Passo Fundo, RS, Brazil; Postgraduate Program in Pharmacology, Federal University of Santa Maria (UFSM), Santa Maria, Brazil; The International Zebrafish Neuroscience Research Consortium (ZNRC), Slidell, LA, USA
| | - Ana C V V Giacomini
- Bioscience Institute, University of Passo Fundo (UPF), Passo Fundo, RS, Brazil; Postgraduate Program in Pharmacology, Federal University of Santa Maria (UFSM), Santa Maria, Brazil; Postgraduate Program in Environmental Sciences, University of Passo Fundo (UPF), Passo Fundo, Brazil
| | - Rodrigo Zanandrea
- Bioscience Institute, University of Passo Fundo (UPF), Passo Fundo, RS, Brazil
| | - Bruna E Dos Santos
- Bioscience Institute, University of Passo Fundo (UPF), Passo Fundo, RS, Brazil
| | - Rafael Genario
- Bioscience Institute, University of Passo Fundo (UPF), Passo Fundo, RS, Brazil
| | | | - Ashton J Friend
- Tulane University School of Science and Engineering, New Orleans, LA, USA
| | - Tamara G Amstislavskaya
- Research Institute of Physiology and Basic Medicine SB RAS, and Department of Neuroscience, Novosibirsk State University, Novosibirsk, Russia
| | - Allan V Kalueff
- School of Pharmacy, Southwest University, Chongqing, China; Ural Federal University, Ekaterinburg, Russia; ZENEREI Research Center, Slidell, LA, USA; Institute of Translational Biomedicine, St. Petersburg State University, St. Petersburg, Russia; Institute of Experimental Medicine, Almazov National Medical Research Center, St. Petersburg, Russia; Russian Research Center for Radiology and Surgical Technologies, Pesochny, Russia; Laboratory of Translational Biopsychiatry, Research Institute of Physiology and Basic Medicine SB RAS, Novosibirsk, Russia.
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27
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Hanyang L, Xuanzhe L, Xuyang C, Yujia Q, Jiarong F, Jun S, Zhihua R. Application of Zebrafish Models in Inflammatory Bowel Disease. Front Immunol 2017; 8:501. [PMID: 28515725 PMCID: PMC5413514 DOI: 10.3389/fimmu.2017.00501] [Citation(s) in RCA: 25] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/10/2017] [Accepted: 04/11/2017] [Indexed: 12/12/2022] Open
Abstract
Inflammatory bowel disease (IBD) is a chronic, recurrent, and remitting inflammatory disease with unclear etiology. As a clinically frequent disease, it can affect individuals throughout their lives, with multiple complications. Unfortunately, traditional murine models are not efficient for the further study of IBD. Thus, effective and convenient animal models are needed. Zebrafish have been used as model organisms to investigate IBD because of their suggested highly genetic similarity to humans and their superiority as laboratory models. The zebrafish model has been used to study the composition of intestinal microbiota, novel genes, and therapeutic approaches. The pathogenesis of IBD is still unclear and many risk factors remain unidentified. In this review, we compare traditional murine models and zebrafish models in terms of advantages, pathogenesis, and drug discovery screening for IBD. We also review the progress and deficiencies of the zebrafish model for scientific applications.
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Affiliation(s)
- Li Hanyang
- Division of Gastroenterology and Hepatology, Key Laboratory of Gastroenterology and Hepatology, Ministry of Health, Inflammatory Bowel Disease Research Center, Shanghai, China.,Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.,Shanghai Institute of Digestive Disease, Shanghai, China
| | - Liu Xuanzhe
- Division of Gastroenterology and Hepatology, Key Laboratory of Gastroenterology and Hepatology, Ministry of Health, Inflammatory Bowel Disease Research Center, Shanghai, China.,Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.,Shanghai Institute of Digestive Disease, Shanghai, China
| | - Chen Xuyang
- Division of Gastroenterology and Hepatology, Key Laboratory of Gastroenterology and Hepatology, Ministry of Health, Inflammatory Bowel Disease Research Center, Shanghai, China.,Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.,Shanghai Institute of Digestive Disease, Shanghai, China
| | - Qiu Yujia
- Division of Gastroenterology and Hepatology, Key Laboratory of Gastroenterology and Hepatology, Ministry of Health, Inflammatory Bowel Disease Research Center, Shanghai, China.,Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.,Shanghai Institute of Digestive Disease, Shanghai, China
| | - Fu Jiarong
- Division of Gastroenterology and Hepatology, Key Laboratory of Gastroenterology and Hepatology, Ministry of Health, Inflammatory Bowel Disease Research Center, Shanghai, China.,Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.,Shanghai Institute of Digestive Disease, Shanghai, China
| | - Shen Jun
- Division of Gastroenterology and Hepatology, Key Laboratory of Gastroenterology and Hepatology, Ministry of Health, Inflammatory Bowel Disease Research Center, Shanghai, China.,Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.,Shanghai Institute of Digestive Disease, Shanghai, China
| | - Ran Zhihua
- Division of Gastroenterology and Hepatology, Key Laboratory of Gastroenterology and Hepatology, Ministry of Health, Inflammatory Bowel Disease Research Center, Shanghai, China.,Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.,Shanghai Institute of Digestive Disease, Shanghai, China
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28
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Løvmo SD, Speth MT, Repnik U, Koppang EO, Griffiths GW, Hildahl JP. Translocation of nanoparticles and Mycobacterium marinum across the intestinal epithelium in zebrafish and the role of the mucosal immune system. DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY 2017; 67:508-518. [PMID: 27343826 DOI: 10.1016/j.dci.2016.06.016] [Citation(s) in RCA: 18] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/14/2016] [Revised: 06/20/2016] [Accepted: 06/20/2016] [Indexed: 06/06/2023]
Abstract
Nano- and microparticles are promising carrier systems for oral delivery of drugs or vaccines, particularly in fish aquaculture. However, the mechanisms of uptake, trans-epithelial transport and immune response to nano/micrometer sized particles, or microorganisms such as bacteria are poorly understood in fish. Here, adult zebrafish were used to study the uptake of different nano- and microparticles and the pathogenic bacteria Mycobacterium marinum in the intestine, and their interactions with epithelial cells and the mucosal immune system. Fluorescent particles or bacteria were delivered directly into the adult zebrafish intestine by oral intubation and their localization was imaged in intestine, liver and spleen sections. Zebrafish do not appear to have M-cells, but both nanoparticles and bacteria were rapidly taken up in the intestine and transported to the liver and spleen. In each tissue, both bacteria and particles largely localized to leukocytes, presumably macrophages.
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Affiliation(s)
- Signe Dille Løvmo
- Department of Biosciences, University of Oslo, Blindernveien 31, 0371 Oslo, Norway
| | - Martin Tobias Speth
- Department of Biosciences, University of Oslo, Blindernveien 31, 0371 Oslo, Norway
| | - Urska Repnik
- Department of Biosciences, University of Oslo, Blindernveien 31, 0371 Oslo, Norway
| | - Erling Olaf Koppang
- School of Veterinary Medicine, Norwegian University of Life Sciences, Ullevålsveien 72, 0454 Oslo, Norway
| | - Gareth Wyn Griffiths
- Department of Biosciences, University of Oslo, Blindernveien 31, 0371 Oslo, Norway.
| | - Jon Paul Hildahl
- Department of Biosciences, University of Oslo, Blindernveien 31, 0371 Oslo, Norway
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29
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Abstract
Although the zebrafish was initially developed as a model system to study embryonic development, it has gained increasing attention as an advantageous system to investigate human diseases, including intestinal disorders. Zebrafish embryos develop rapidly, and their digestive system is fully functional and visible by 5days post fertilization. There is a large degree of homology between the intestine of zebrafish and higher vertebrate organisms in terms of its cellular composition and function as both a digestive and immune organ. Furthermore, molecular pathways regulating injury and immune responses are highly conserved. In this chapter, we provide an overview of studies addressing developmental and physiological processes relevant to human intestinal disease. These studies include those related to congenital disorders, host-microbiota interactions, inflammatory diseases, motility disorders, and intestinal cancer. We also highlight the utility of zebrafish to functionally validate candidate genes identified through mutational analyses and genome-wide association studies, and discuss methodologies to investigate the intestinal biology that are unique to zebrafish.
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Affiliation(s)
- X Zhao
- University of Pennsylvania, Philadelphia, PA, United States
| | - M Pack
- University of Pennsylvania, Philadelphia, PA, United States
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30
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Abstract
Zebrafish are an excellent model for the study of intestinal immunity. The availability of several transgenic reporter fish for different innate and adaptive immune cells and the high homology in terms of gut function and morphology enables in depth analysis of the process of intestinal inflammation. Here, we describe a method to induce intestinal inflammation by intra-rectal injection of the hapten oxazolone in adult zebrafish.
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Affiliation(s)
- Sylvia Brugman
- Animal Sciences Group, Cell Biology and Immunology, Wageningen University, De Elst 1, 6708, WD, Wageningen, The Netherlands.
| | - Edward E S Nieuwenhuis
- Department of Pediatrics, Wilhelmina Children's Hospital Utrecht, University Medical Center Utrecht, Utrecht, The Netherlands
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31
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Brugman S. The zebrafish as a model to study intestinal inflammation. DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY 2016; 64:82-92. [PMID: 26902932 DOI: 10.1016/j.dci.2016.02.020] [Citation(s) in RCA: 177] [Impact Index Per Article: 19.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 09/23/2015] [Revised: 02/16/2016] [Accepted: 02/17/2016] [Indexed: 06/05/2023]
Abstract
Starting out as a model for developmental biology, during the last decade, zebrafish have also gained the attention of the immunologists and oncologists. Due to its small size, high fecundity and full annotation of its genome, the zebrafish is an attractive model system. The fact that fish are transparent early in life combined with the growing list of immune cell reporter fish, enables in vivo tracking of immune responses in a complete organism. Since zebrafish develop ex utero from a fertilized egg, immune development can be monitored from the start of life. Given that several gut functions and immune genes are conserved between zebrafish and mammals, the zebrafish is an interesting model organism to investigate fundamental processes underlying intestinal inflammation and injury. This review will first provide some background on zebrafish intestinal development, bacterial colonization and immunity, showing the similarities and differences compared to mammals. This will be followed by an overview of the existing models for intestinal disease, and concluded by future perspectives in light of the newest technologies and insights.
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Affiliation(s)
- Sylvia Brugman
- Animal Sciences Group, Cell Biology and Immunology, Wageningen University, De Elst 1, room Ee1253, 6708 WD Wageningen, Netherlands.
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32
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Calduch-Giner JA, Sitjà-Bobadilla A, Pérez-Sánchez J. Gene Expression Profiling Reveals Functional Specialization along the Intestinal Tract of a Carnivorous Teleostean Fish (Dicentrarchus labrax). Front Physiol 2016; 7:359. [PMID: 27610085 PMCID: PMC4997091 DOI: 10.3389/fphys.2016.00359] [Citation(s) in RCA: 33] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/14/2016] [Accepted: 08/05/2016] [Indexed: 01/23/2023] Open
Abstract
High-quality sequencing reads from the intestine of European sea bass were assembled, annotated by similarity against protein reference databases and combined with nucleotide sequences from public and private databases. After redundancy filtering, 24,906 non-redundant annotated sequences encoding 15,367 different gene descriptions were obtained. These annotated sequences were used to design a custom, high-density oligo-microarray (8 × 15 K) for the transcriptomic profiling of anterior (AI), middle (MI), and posterior (PI) intestinal segments. Similar molecular signatures were found for AI and MI segments, which were combined in a single group (AI-MI) whereas the PI outstood separately, with more than 1900 differentially expressed genes with a fold-change cutoff of 2. Functional analysis revealed that molecular and cellular functions related to feed digestion and nutrient absorption and transport were over-represented in AI-MI segments. By contrast, the initiation and establishment of immune defense mechanisms became especially relevant in PI, although the microarray expression profiling validated by qPCR indicated that these functional changes are gradual from anterior to posterior intestinal segments. This functional divergence occurred in association with spatial transcriptional changes in nutrient transporters and the mucosal chemosensing system via G protein-coupled receptors. These findings contribute to identify key indicators of gut functions and to compare different fish feeding strategies and immune defense mechanisms acquired along the evolution of teleosts.
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Affiliation(s)
- Josep A Calduch-Giner
- Nutrigenomics and Fish Growth Endocrinology Group, Biology, Culture and Pathology of Marine Species, Institute of Aquaculture Torre de la Sal (IATS-CSIC) Castellón, Spain
| | - Ariadna Sitjà-Bobadilla
- Fish Pathology Group, Biology, Culture and Pathology of Marine Species, Institute of Aquaculture Torre de la Sal (IATS-CSIC) Castellón, Spain
| | - Jaume Pérez-Sánchez
- Nutrigenomics and Fish Growth Endocrinology Group, Biology, Culture and Pathology of Marine Species, Institute of Aquaculture Torre de la Sal (IATS-CSIC) Castellón, Spain
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33
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Morales Fénero CI, Colombo Flores AA, Câmara NOS. Inflammatory diseases modelling in zebrafish. World J Exp Med 2016; 6:9-20. [PMID: 26929916 PMCID: PMC4759353 DOI: 10.5493/wjem.v6.i1.9] [Citation(s) in RCA: 17] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/29/2015] [Revised: 10/20/2015] [Accepted: 12/18/2015] [Indexed: 02/06/2023] Open
Abstract
The ingest of diets with high content of fats and carbohydrates, low or no physical exercise and a stressful routine are part of the everyday lifestyle of most people in the western world. These conditions are triggers for different diseases with complex interactions between the host genetics, the metabolism, the immune system and the microbiota, including inflammatory bowel diseases (IBD), obesity and diabetes. The incidence of these disorders is growing worldwide; therefore, new strategies for its study are needed. Nowadays, the majority of researches are in use of murine models for understand the genetics, physiopathology and interaction between cells and signaling pathways to find therapeutic solutions to these diseases. The zebrafish, a little tropical water fish, shares 70% of our genes and conserves anatomic and physiological characteristics, as well as metabolical pathways, with mammals, and is rising as a new complementary model for the study of metabolic and inflammatory diseases. Its high fecundity, fast development, transparency, versatility and low cost of maintenance makes the zebrafish an interesting option for new researches. In this review, we offer a discussion of the existing genetic and induced zebrafish models of two important Western diseases that have a strong inflammatory component, the IBD and the obesity.
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34
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Katzenback BA. Antimicrobial Peptides as Mediators of Innate Immunity in Teleosts. BIOLOGY 2015; 4:607-39. [PMID: 26426065 PMCID: PMC4690011 DOI: 10.3390/biology4040607] [Citation(s) in RCA: 78] [Impact Index Per Article: 7.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 08/31/2015] [Revised: 09/16/2015] [Accepted: 09/17/2015] [Indexed: 12/16/2022]
Abstract
Antimicrobial peptides (AMPs) have been identified throughout the metazoa suggesting their evolutionarily conserved nature and their presence in teleosts is no exception. AMPs are short (18–46 amino acids), usually cationic, amphipathic peptides. While AMPs are diverse in amino acid sequence, with no two AMPs being identical, they collectively appear to have conserved functions in the innate immunity of animals towards the pathogens they encounter in their environment. Fish AMPs are upregulated in response to pathogens and appear to have direct broad-spectrum antimicrobial activity towards both human and fish pathogens. However, an emerging role for AMPs as immunomodulatory molecules has become apparent—the ability of AMPs to activate the innate immune system sheds light onto the multifaceted capacity of these small peptides to combat pathogens through direct and indirect means. Herein, this review focuses on the role of teleost AMPs as modulators of the innate immune system and their regulation in response to pathogens or other exogenous molecules. The capacity to regulate AMP expression by exogenous factors may prove useful in modulating AMP expression in fish to prevent disease, particularly in aquaculture settings where crowded conditions and environmental stress pre-dispose these fish to infection.
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Affiliation(s)
- Barbara A Katzenback
- Department of Biology, University of Waterloo, 200 University Ave West, Waterloo, ON N2L 3G1, Canada.
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35
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Chew TS, O'Shea NR, Sewell GW, Oehlers SH, Mulvey CM, Crosier PS, Godovac-Zimmermann J, Bloom SL, Smith AM, Segal AW. Optineurin deficiency in mice contributes to impaired cytokine secretion and neutrophil recruitment in bacteria-driven colitis. Dis Model Mech 2015; 8:817-29. [PMID: 26044960 PMCID: PMC4527293 DOI: 10.1242/dmm.020362] [Citation(s) in RCA: 42] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/18/2015] [Accepted: 05/11/2015] [Indexed: 12/16/2022] Open
Abstract
Crohn's disease (CD) is associated with delayed neutrophil recruitment and bacterial clearance at sites of acute inflammation as a result of impaired secretion of proinflammatory cytokines by macrophages. To investigate the impaired cytokine secretion and confirm our previous findings, we performed transcriptomic analysis in macrophages and identified a subgroup of individuals with CD who had low expression of the autophagy receptor optineurin (OPTN). We then clarified the role of OPTN deficiency in: macrophage cytokine secretion; mouse models of bacteria-driven colitis and peritonitis; and zebrafish Salmonella infection. OPTN-deficient bone-marrow-derived macrophages (BMDMs) stimulated with heat-killed Escherichia coli secreted less proinflammatory TNFα and IL6 cytokines despite similar gene transcription, which normalised with lysosomal and autophagy inhibitors, suggesting that TNFα is mis-trafficked to lysosomes via bafilomycin-A-dependent pathways in the absence of OPTN. OPTN-deficient mice were more susceptible to Citrobacter colitis and E. coli peritonitis, and showed reduced levels of proinflammatory TNFα in serum, diminished neutrophil recruitment to sites of acute inflammation and greater mortality, compared with wild-type mice. Optn-knockdown zebrafish infected with Salmonella also had higher mortality. OPTN plays a role in acute inflammation and neutrophil recruitment, potentially via defective macrophage proinflammatory cytokine secretion, which suggests that diminished OPTN expression in humans might increase the risk of developing CD.
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Affiliation(s)
- Thean S Chew
- Division of Medicine, University College London, London, WC1E 6JF, UK
| | - Nuala R O'Shea
- Division of Medicine, University College London, London, WC1E 6JF, UK
| | - Gavin W Sewell
- Division of Medicine, University College London, London, WC1E 6JF, UK
| | - Stefan H Oehlers
- Department of Molecular Medicine and Pathology, University of Auckland, Auckland 1001, New Zealand
| | - Claire M Mulvey
- Division of Medicine, University College London, London, WC1E 6JF, UK Department of Genetics, University of Cambridge, Cambridge, CB2 3EH, UK
| | - Philip S Crosier
- Department of Molecular Medicine and Pathology, University of Auckland, Auckland 1001, New Zealand
| | | | - Stuart L Bloom
- Department of Gastroenterology, University College London Hospital, London, NW1 2BU, UK
| | - Andrew M Smith
- Division of Medicine, University College London, London, WC1E 6JF, UK Microbial Diseases, Eastman Dental Institute, University College London, London, WC1X 8LD, UK
| | - Anthony W Segal
- Division of Medicine, University College London, London, WC1E 6JF, UK
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Jheng YH, Lee LH, Ting CH, Pan CY, Hui CF, Chen JY. Zebrafish fed on recombinant Artemia expressing epinecidin-1 exhibit increased survival and altered expression of immunomodulatory genes upon Vibrio vulnificus infection. FISH & SHELLFISH IMMUNOLOGY 2015; 42:1-15. [PMID: 25462461 DOI: 10.1016/j.fsi.2014.10.019] [Citation(s) in RCA: 21] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 07/19/2014] [Revised: 10/13/2014] [Accepted: 10/15/2014] [Indexed: 06/04/2023]
Abstract
Artemia has been used extensively in aquaculture as fodder for larval fish, shrimp, and shellfish. Epinecidin-1, an antimicrobial peptide, was isolated from grouper (Epinephelus coioides) in 2005. Epinecidin-1 has been previously reported to possess antimicrobial activity against several Gram-positive and Gram-negative bacterial species, including Staphylococcus coagulase, Pseudomonas aeruginosa, Streptococcus pyogenes, and Vibrio vulnificus. In this study, we used electroporation to introduce plasmid DNA encoding a green fluorescent protein (EGFP)-epinecidin-1 fusion protein under the control of the cytomegalovirus (CMV) promoter into decapsulated Artemia cysts. Optimization of various properties (including cyst weight (0.2 g), plasmid concentration (50 μg/100 μl), and pulse voltage (150 V), length (10 ms), and number (2)) resulted in a hatching rate of 41.15%, a transfection efficiency of 49.81%, and a fluorescence intensity (A.U.) of 47.46. The expression of EGFP-epinecidin-1 was first detected by quantitative RT-PCR at 120 h post-electroporation, and protein was identified by Western blot at the same time. Furthermore, the EGFP-epinecidin-1 protein inhibited V. vulnificus (204) growth, as demonstrated by zone of inhibition studies. Zebrafish fed on transgenic Artemia expressing CMV-gfp-epi combined with commercial fodder were more resistant to infection by V. vulnificus (204): survival rate was enhanced by over 70% at 7, 14, and 21 days post-infection, and bacterial numbers in the liver and intestine were reduced. In addition, feeding of transgenic Artemia to zebrafish affected the immunomodulatory response to V. vulnificus (204) infection; expression of immune-responsive genes, including hepcidin and defbl2, was altered, as shown by qPCR. These findings suggest that feeding transgenic Artemia expressing CMV-gfp-epi to larval fish has antimicrobial effects, without the drawbacks of introducing drug residues or inducing bacterial drug resistance.
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Affiliation(s)
- Yu-Hsuan Jheng
- Institute of Bioscience and Biotechnology, National Taiwan Ocean University, 2, Pei Ning Road, Keelung 20224, Taiwan
| | - Lin-Han Lee
- Marine Research Station, Institute of Cellular and Organismic Biology, Academia Sinica, 23-10 Dahuen Road, Jiaushi, Ilan 262, Taiwan
| | - Chen-Hung Ting
- Marine Research Station, Institute of Cellular and Organismic Biology, Academia Sinica, 23-10 Dahuen Road, Jiaushi, Ilan 262, Taiwan
| | - Chieh-Yu Pan
- Department and Graduate Institute of Aquaculture, National Kaohsiung Marine University, Kaohsiung, Taiwan 811, Taiwan
| | - Cho-Fat Hui
- Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan
| | - Jyh-Yih Chen
- Institute of Bioscience and Biotechnology, National Taiwan Ocean University, 2, Pei Ning Road, Keelung 20224, Taiwan; Marine Research Station, Institute of Cellular and Organismic Biology, Academia Sinica, 23-10 Dahuen Road, Jiaushi, Ilan 262, Taiwan.
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37
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Gratacap RL, Bergeron AC, Wheeler RT. Modeling mucosal candidiasis in larval zebrafish by swimbladder injection. J Vis Exp 2014:e52182. [PMID: 25490695 DOI: 10.3791/52182] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/22/2022] Open
Abstract
Early defense against mucosal pathogens consists of both an epithelial barrier and innate immune cells. The immunocompetency of both, and their intercommunication, are paramount for the protection against infections. The interactions of epithelial and innate immune cells with a pathogen are best investigated in vivo, where complex behavior unfolds over time and space. However, existing models do not allow for easy spatio-temporal imaging of the battle with pathogens at the mucosal level. The model developed here creates a mucosal infection by direct injection of the fungal pathogen, Candida albicans, into the swimbladder of juvenile zebrafish. The resulting infection enables high-resolution imaging of epithelial and innate immune cell behavior throughout the development of mucosal disease. The versatility of this method allows for interrogation of the host to probe the detailed sequence of immune events leading to phagocyte recruitment and to examine the roles of particular cell types and molecular pathways in protection. In addition, the behavior of the pathogen as a function of immune attack can be imaged simultaneously by using fluorescent protein-expressing C. albicans. Increased spatial resolution of the host-pathogen interaction is also possible using the described rapid swimbladder dissection technique. The mucosal infection model described here is straightforward and highly reproducible, making it a valuable tool for the study of mucosal candidiasis. This system may also be broadly translatable to other mucosal pathogens such as mycobacterial, bacterial or viral microbes that normally infect through epithelial surfaces.
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Affiliation(s)
- Remi L Gratacap
- Department of Molecular and Biomedical Sciences, University of Maine
| | - Audrey C Bergeron
- Department of Molecular and Biomedical Sciences, University of Maine
| | - Robert T Wheeler
- Department of Molecular and Biomedical Sciences, University of Maine; Graduate School of Biomedical Sciences and Engineering, University of Maine;
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38
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Liu X, Wu H, Chang X, Tang Y, Liu Q, Zhang Y. Notable mucosal immune responses induced in the intestine of zebrafish (Danio rerio) bath-vaccinated with a live attenuated Vibrio anguillarum vaccine. FISH & SHELLFISH IMMUNOLOGY 2014; 40:99-108. [PMID: 24997435 DOI: 10.1016/j.fsi.2014.06.030] [Citation(s) in RCA: 47] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/12/2014] [Revised: 06/10/2014] [Accepted: 06/13/2014] [Indexed: 06/03/2023]
Abstract
Live attenuated vaccine is one of the efficient vaccine candidates in aquaculture, which can be easily delivered to fish via bath-vaccination. An outstanding advantage of bath-vaccination is that vaccine delivery is through the same route as that utilized by many fish pathogens, generating specific mucosal immune responses. In this work, we investigated the mucosal immune responses induced by a live attenuated Vibrio anguillarum vaccine in zebrafish via bath-vaccination. Bacteria proliferated rapidly in 3 h after vaccination and maintained at a high level until 6 h in the intestine. Besides, bacteria persisted in the intestine for a longer time whereas decreased rapidly in the skin and gills. Moreover, a significant up-regulation of TLR5 triggering a MyD88-dependent signaling pathway was observed in the intestine, which implied that flagella were the crucial antigenic component of the live attenuated vaccine. And macrophages and neutrophils showed active responses participating in antigen recognition and sampling after vaccination. Furthermore, an inflammation was observed with plenty of lymphocytes in the intestine at 24 h post vaccination but eliminated within 7 days. In conclusion, the live attenuated V. anguillarum vaccine induced notable mucosal immune responses in the intestine which could be used as a mucosal vaccine vector in the future.
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Affiliation(s)
- Xiaohong Liu
- State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China
| | - Haizhen Wu
- State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China.
| | - Xinyue Chang
- State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China
| | - Yufei Tang
- State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China
| | - Qin Liu
- State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China
| | - Yuanxing Zhang
- State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China
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39
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De Deken X, Corvilain B, Dumont JE, Miot F. Roles of DUOX-mediated hydrogen peroxide in metabolism, host defense, and signaling. Antioxid Redox Signal 2014; 20:2776-93. [PMID: 24161126 DOI: 10.1089/ars.2013.5602] [Citation(s) in RCA: 87] [Impact Index Per Article: 7.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
SIGNIFICANCE Among the NADPH oxidases, the dual oxidases, DUOX1 and DUOX2, constitute a distinct subfamily initially called thyroid oxidases, based on their high level of expression in thyroid tissue. Genetic alterations causing inherited hypothyroidism clearly demonstrate their physiological implication in thyroid hormonogenesis. However, a growing list of biological functions triggered by DUOX-dependent reactive oxygen species (ROS) in highly differentiated mucosae have recently emerged. RECENT ADVANCES A role of DUOX enzymes as ROS providers for lactoperoxidase-mediated killing of invading pathogens has been well established and a role in bacteria chemorepulsion has been proposed. Control of DUOX expression and activity by inflammatory molecules and immune receptor activation consolidates their contributions to innate immune defense of mucosal surfaces. Recent studies conducted in ancestral organisms have identified effectors of DUOX redox signaling involved in wound healing including epithelium regeneration and leukocyte recruitment. Moreover, local generation of hydrogen peroxide (H2O2) by DUOX has also been suggested to constitute a positive feedback loop to promote receptor signaling activation. CRITICAL ISSUES A correct balance between H2O2 generation and detoxification mechanisms must be properly maintained to avoid oxidative damages. Overexpression of DUOX genes has been associated with an increasing number of chronic inflammatory diseases. Furthermore, H2O2-mediated DNA damage supports a mutagenic function promoting tumor development. FUTURE DIRECTIONS Despite the high sequence similarity shared between DUOX1 and DUOX2, the two isoforms present distinct regulations, tissue expression and catalytic functions. The phenotypic characterization of novel DUOX/DUOXA invalidated animal models will be very useful for defining their medical importance in pathological conditions.
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Affiliation(s)
- Xavier De Deken
- Faculté de Médecine, Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire (IRIBHM), Université Libre de Bruxelles (ULB) , Brussels, Belgium
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40
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Abstract
Understanding a complex pathology such as inflammatory bowel disease, where host genetics (innate and adaptive immunity, barrier function) and environmental factors (microbes, diet, and stress) interact together to influence disease onset and severity, requires multipronged approaches to model these numerous variables. Researchers have typically relied on preclinical models of mouse and rat origin to push the boundary of knowledge further. However, incorporation of novel vertebrate models may contribute to new knowledge on specific aspects of intestinal homeostasis. An emerging literature has seen the use of zebrafish as a novel animal system to study key aspects of host-microbe interactions in the intestine. In this review, we briefly introduce components of host-microbiota interplay in the developing zebrafish intestine and summarize key lessons learned from this animal system; review important chemically induced and genetically engineered zebrafish models of intestinal immune disorders; and discuss perspectives and limitations of the zebrafish model system.
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Affiliation(s)
- Ye Yang
- Department of Medicine, University of Florida, Gainesville, Florida
| | - Sarah Tomkovich
- Department of Medicine, University of Florida, Gainesville, Florida,Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina
| | - Christian Jobin
- Department of Medicine, University of Florida, Gainesville, Florida,Department of Infectious Diseases and Pathology, University of Florida, Gainesville, Florida
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41
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Zhang H, Shen B, Wu H, Gao L, Liu Q, Wang Q, Xiao J, Zhang Y. Th17-like immune response in fish mucosal tissues after administration of live attenuated Vibrio anguillarum via different vaccination routes. FISH & SHELLFISH IMMUNOLOGY 2014; 37:229-238. [PMID: 24561130 DOI: 10.1016/j.fsi.2014.02.007] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/24/2013] [Revised: 02/06/2014] [Accepted: 02/12/2014] [Indexed: 06/03/2023]
Abstract
This study aimed to investigate the protective mucosal immunity elicited by live attenuated Vibrio anguillarum in fish. Zebrafish were immunized by bath or injection way, and undertook bath challenge at 28 days post vaccination. The results implied that bath vaccination was the better delivery route for inducing the protective immunity against bath challenge in zebrafish. The expressions of genes related to Th1, Th2 and Th17 cells were measured in the mucosal tissues of vaccinated and challenged zebrafish. Gene expression profiles showed that Th17-like responses were induced in mucosal immune system by vaccination via bath and injection routes while Th1 and Th2-like responses were not remarkable. Compared to injection vaccination, bath vaccination elicited the intense Th17-like immune responses in the gut tissue of zebrafish. Additionally, in gills and skin, Th17-like mucosal immunity elicited by injection vaccination occurred later than that by bath vaccination. Our results proved the immunological importance of gut in bath vaccination and the presence of two-compartmental model for immune response in zebrafish. In conclusion, bath vaccination more efficiently elicited protective Th17-like immunity than injection vaccination in mucosal tissues of vaccinated zebrafish. In turbot, effective immune protection against wild-type V. anguillarum was obtained by bath-vaccinated and the Th17-like responses were found in mucosal and systemic tissues.
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Affiliation(s)
- Hua Zhang
- State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
| | - Binbing Shen
- State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
| | - Haizhen Wu
- State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
| | - Liang Gao
- State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
| | - Qin Liu
- State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
| | - Qiyao Wang
- State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
| | - Jingfan Xiao
- State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
| | - Yuanxing Zhang
- State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
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42
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Thakur PC, Davison JM, Stuckenholz C, Lu L, Bahary N. Dysregulated phosphatidylinositol signaling promotes endoplasmic-reticulum-stress-mediated intestinal mucosal injury and inflammation in zebrafish. Dis Model Mech 2013; 7:93-106. [PMID: 24135483 PMCID: PMC3882052 DOI: 10.1242/dmm.012864] [Citation(s) in RCA: 33] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/11/2022] Open
Abstract
Dysregulated phosphatidylinositol (PI) signaling has been implicated in human gastrointestinal (GI) malignancies and inflammatory states, underlining the need to study pathophysiological roles of PI in an in vivo genetic model. Here, we study the significance of PI in GI pathophysiology using the zebrafish mutant cdipthi559, which lacks PI synthesis, and unravel a crucial role of PI in intestinal mucosal integrity and inflammation. The cdipthi559 mutants exhibit abnormal villous architecture and disorganized proliferation of intestinal epithelial cells (IECs), with pathologies reminiscent of inflammatory bowel disease (IBD), including apoptosis of goblet cells, abnormal mucosecretion, bacterial overgrowth and leukocyte infiltration. The mutant IECs exhibit vacuolation, microvillus atrophy and impaired proliferation. The cdipthi559 gene expression profile shows enrichment of acute phase response signaling, and the endoplasmic reticulum (ER) stress factors hspa5 and xbp1 are robustly activated in the mutant GI tissue. Temporal electron micrographic analyses reveal that PI-deficient IECs undergo sequential ER-Golgi disruption, mitochondrial depletion, macroautophagy and cell death, consistent with chronic ER-stress-mediated cytopathology. Furthermore, pharmacological induction of ER stress by inhibiting protein glycosylation or PI synthase inhibition in leukocyte-specific reporter lines replicates the cdipthi559 inflammatory phenotype, suggesting a fundamental role of PI metabolism and ER stress in mucosal inflammation. Antibiotics and anti-inflammatory drugs resolved the inflammation, but not the autophagic necroapoptosis of IECs, suggesting that bacterial overgrowth can exacerbate ER stress pathology, whereas persistent ER stress is sufficient to trigger inflammation. Interestingly, the intestinal phenotype was partially alleviated by chemical chaperones, suggesting their therapeutic potential. Using zebrafish genetic and pharmacological models, this study demonstrates a newly identified link between intracellular PI signaling and ER-stress-mediated mucosal inflammation. The zebrafish cdipt mutants provide a powerful tool for dissecting the fundamental mechanisms of ER-stress-mediated human GI diseases and a platform to develop molecularly targeted therapies.
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Affiliation(s)
- Prakash C Thakur
- Department of Medicine, Division of Hematology/Oncology, University of Pittsburgh Cancer Institute, Pittsburgh, PA 15232, USA
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43
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Gratacap RL, Rawls JF, Wheeler RT. Mucosal candidiasis elicits NF-κB activation, proinflammatory gene expression and localized neutrophilia in zebrafish. Dis Model Mech 2013; 6:1260-70. [PMID: 23720235 PMCID: PMC3759345 DOI: 10.1242/dmm.012039] [Citation(s) in RCA: 51] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022] Open
Abstract
The epithelium performs a balancing act at the interface between an animal and its environment to enable both pathogen killing and tolerance of commensal microorganisms. Candida albicans is a clinically important human commensal that colonizes all human mucosal surfaces, yet is largely prevented from causing mucosal infections in immunocompetent individuals. Despite the importance of understanding host-pathogen interactions at the epithelium, no immunocompetent vertebrate model has been used to visualize these dynamics non-invasively. Here we demonstrate important similarities between swimbladder candidiasis in the transparent zebrafish and mucosal infection at the mammalian epithelium. Specifically, in the zebrafish swimmbladder infection model, we show dimorphic fungal growth, both localized and tissue-wide epithelial NF-κB activation, induction of NF-κB -dependent proinflammatory genes, and strong neutrophilia. Consistent with density-dependence models of host response based primarily on tissue culture experiments, we show that only high-level infection provokes widespread activation of NF-κB in epithelial cells and induction of proinflammatory genes. Similar to what has been found using in vitro mammalian models, we find that epithelial NF-κB activation can occur at a distance from the immediate site of contact with epithelial cells. Taking advantage of the ability to non-invasively image infection and host signaling at high resolution, we also report that epithelial NF-κB activation is diminished when phagocytes control the infection. This is the first system to model host response to mucosal infection in the juvenile zebrafish, and offers unique opportunities to investigate the tripartite interactions of C. albicans, epithelium and immune cells in an intact host.
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Affiliation(s)
- Remi L Gratacap
- Department of Molecular and Biomedical Sciences, University of Maine, Orono, ME 04469, USA
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44
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A novel beta-defensin antimicrobial peptide in Atlantic cod with stimulatory effect on phagocytic activity. PLoS One 2013; 8:e62302. [PMID: 23638029 PMCID: PMC3636224 DOI: 10.1371/journal.pone.0062302] [Citation(s) in RCA: 44] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/26/2012] [Accepted: 03/22/2013] [Indexed: 11/19/2022] Open
Abstract
A novel defensin antimicrobial peptide gene was identified in Atlantic cod, Gadus morhua. This three exon/two intron defensin gene codes for a peptide precursor consisting of two domains: a signal peptide of 26 amino acids and a mature peptide of 40 residues. The mature cod defensin has six conserved cysteine residues that form 1–5, 2–4 and 3–6 disulphide bridges. This pattern is typical of beta-defensins and this gene was therefore named cod beta-defensin (defb). The tertiary structure of Defb exhibits an α/β fold with one α helix and β1β2β3 sheets. RT-PCR analysis indicated that defb transcripts were present mainly in the swim bladder and peritoneum wall but could also be detected at moderate to low levels in skin, head- and excretory kidneys. In situ hybridisation revealed that defb was specifically expressed by cells located in the swim bladder submucosa and the oocytes. During embryonic development, defb gene transcripts were detectable from the golden eye stage onwards and their expression was restricted to the swim bladder and retina. Defb was differentially expressed in several tissues following antigenic challenge with Vibrio anguillarum, being up-regulated up to 25-fold in head kidney. Recombinant Defb displayed antibacterial activity, with a minimal inhibitory concentration of 0.4–0.8 µM and 25–50 µM against the Gram-(+) bacteria Planococcus citreus and Micrococcus luteus, respectively. In addition, Defb stimulated phagocytic activity of cod head kidney leucocytes invitro. These findings imply that beta-defensins may play an important role in the innate immune response of Atlantic cod.
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45
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Oehlers SH, Flores MV, Hall CJ, Okuda KS, Sison JO, Crosier KE, Crosier PS. Chemically induced intestinal damage models in zebrafish larvae. Zebrafish 2013; 10:184-93. [PMID: 23448252 DOI: 10.1089/zeb.2012.0824] [Citation(s) in RCA: 69] [Impact Index Per Article: 5.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/30/2022] Open
Abstract
Several intestinal damage models have been developed using zebrafish, with the aim of recapitulating aspects of human inflammatory bowel disease (IBD). These experimentally induced inflammation models have utilized immersion exposure to an array of colitogenic agents (including live bacteria, bacterial products, and chemicals) to induce varying severity of inflammation. This technical report describes methods used to generate two chemically induced intestinal damage models using either dextran sodium sulfate (DSS) or trinitrobenzene sulfonic acid (TNBS). Methods to monitor intestinal damage and inflammatory processes, and chemical-genetic methods to manipulate the host response to injury are also described.
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Affiliation(s)
- Stefan H Oehlers
- Department of Molecular Medicine and Pathology, School of Medical Sciences, The University of Auckland, Auckland, New Zealand
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46
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Casadei E, Bird S, Vecino JLG, Wadsworth S, Secombes CJ. The effect of peptidoglycan enriched diets on antimicrobial peptide gene expression in rainbow trout (Oncorhynchus mykiss). FISH & SHELLFISH IMMUNOLOGY 2013; 34:529-537. [PMID: 23220715 DOI: 10.1016/j.fsi.2012.11.027] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/25/2012] [Revised: 11/26/2012] [Accepted: 11/27/2012] [Indexed: 06/01/2023]
Abstract
The aim of this study was to investigate the effect of feeding rainbow trout (Oncorhynchus mykiss) peptidoglycan (PG) enriched diets on antimicrobial peptide (AMP) gene expression. Fish were divided into 5 groups and fed diets containing 0, 5, 10, 50 and 100 mg PG/Kg, and sampled 1, 7 and 14 days later. The expression of eight AMP genes (four defensins, two cathelicidins and two liver expressed AMPs) was determined in skin, gill, gut and liver, tissues important for first lines of defence or production of acute phase proteins. Up-regulation of many AMPs was found after feeding the PG enriched diets, with sequential expression seen over the time course studied, where defensins were typically expressed early and cathelicidins and LEAPs later on. A number of clear differences in AMP responsiveness between the tissues examined were also apparent. Of the four PG concentrations used, 5 mg PG/Kg did not always elicit AMP gene induction or to the same degree as seen with the other diets. The three higher dose groups generally showed similar trends although differences in fold change were more pronounced in the 50 and 100 mg PG/Kg groups. Curiously several AMPs were down-regulated after 14 days of feeding in gills, gut and liver. Nevertheless, overall the PG enriched diets had a positive effect on AMP expression. Further investigations now need to be undertaken to confirm whether this higher AMP gene expression correlates with protection against common bacterial diseases and if PG enriched diets have value as a means to temporarily boost the piscine immune system.
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Affiliation(s)
- Elisa Casadei
- Scottish Fish Immunology Research Centre, Institute of Biological and Environmental Sciences, University of Aberdeen, Aberdeen, UK
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47
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Cantas L, Sørby JRT, Aleström P, Sørum H. Culturable gut microbiota diversity in zebrafish. Zebrafish 2012; 9:26-37. [PMID: 22428747 DOI: 10.1089/zeb.2011.0712] [Citation(s) in RCA: 62] [Impact Index Per Article: 4.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
Abstract
The zebrafish (Danio rerio) is an increasingly used laboratory animal model in basic biology and biomedicine, novel drug development, and toxicology. The wide use has increased the demand for optimized husbandry protocols to ensure animal health care and welfare. The knowledge about the correlation between culturable zebrafish intestinal microbiota and health in relation to environmental factors and management procedures is very limited. A semi-quantitative level of growth of individual types of bacteria was determined and associated with sampling points. A total of 72 TAB line zebrafish from four laboratories (Labs A-D) in the Zebrafish Network Norway were used. Diagnostic was based on traditional bacterial culture methods and biochemical characterization using commercial kits, followed by 16S rDNA gene sequencing from pure subcultures. Also selected Gram-negative isolates were analyzed for antibiotic susceptibility to 8 different antibiotics. A total of 13 morphologically different bacterial species were the most prevalent: Aeromonas hydrophila, Aeromonas sobria, Vibrio parahaemolyticus, Photobacterium damselae, Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas luteola, Comamonas testosteroni, Ochrobactrum anthropi, Staphylococcus cohnii, Staphylococcus epidermidis, Staphylococcus capitis, and Staphylococcus warneri. Only Lab B had significantly higher levels of total bacterial growth (OR=2.03), whereas numbers from Lab C (OR=1.01) and Lab D (OR=1.12) were found to be similar to the baseline Lab A. Sexually immature individuals had a significantly higher level of harvested total bacterial growth than mature fish (OR=0.82), no statistically significant differences were found between male and female fish (OR=1.01), and the posterior intestinal segment demonstrated a higher degree of culturable bacteria than the anterior segment (OR=4.1). Multiple antibiotic (>3) resistance was observed in 17% of the strains. We propose that a rapid conventional diagnostic bacteriological assay on the culturable microbiota profiles can be designed and used as quality measure of the husbandry routines of a zebrafish facility to ensure a bacterial standard safeguarding the zebrafish health and welfare.
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Affiliation(s)
- Leon Cantas
- Department of Food Safety and Infection Biology, Norwegian School of Veterinary Science, Oslo, Norway.
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48
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Ruangsri J, Fernandes JMO, Rombout JHWM, Brinchmann MF, Kiron V. Ubiquitous presence of piscidin-1 in Atlantic cod as evidenced by immunolocalisation. BMC Vet Res 2012; 8:46. [PMID: 22538079 PMCID: PMC3395576 DOI: 10.1186/1746-6148-8-46] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/08/2011] [Accepted: 04/26/2012] [Indexed: 11/25/2022] Open
Abstract
Background Antimicrobial peptides (AMPs), the natural antibiotics bestowed upon all forms of life, consist of small molecular weight proteins with a broad spectrum antimicrobial activity against a variety of pathogenic microorganisms. Piscidins are one of the AMP families that are imperative for the innate defence mechanisms of teleosts. Atlantic cod, a basal fish belonging to the superorder Paracanthopterygii also possesses multiple piscidin peptides. Two piscidin paralogues (pis1 and pis2) and a novel alternative splice variant of pis2 of this fish were previously described by us. To shed light on other potent roles of these molecules, now we have mapped the distribution of piscidin 1 (Pis1), in different tissues and organs of cod through immunohistochemistry (IHC) employing an affinity purified polyclonal antibody specific to Pis1. Results Various cell types and tissues of Atlantic cod including those from the immune organs of naïve fish are armed with Pis1 peptide. Different types of the blood leucocytes and phagocytic cells among the leucocytes examined gave a relatively strong indication of Pis1 immunopositivity. In addition, other cell types such as hematopoietic cells, epithelial cells and multi-granular cells located in the mucosal and hematopoietic tissues were also Pis1-immunoreactive. More interestingly, chondrocytes appear to produce Pis1 and this is the first report on the presence of an AMP in cartilage tissue of fish. Furthermore, Pis1 immunopositivity was detected in other tissues and organs of naïve fish including neural tissues, exocrine and endocrine glands, compound gland cells, excretory kidney, intestinal and respiratory epithelial cells, swim bladder, skin and hypodermis layer, myosepta, liver, heart, eye and oocytes. Conclusions Pis1 peptide is produced by various cell types located in different tissues and organs of Atlantic cod. It is present in all immune-related organs of naïve fish and the elevated peptide expression following phagocytosis strongly suggest their involvement in innate defence. Further, its widespread occurrence in non-immune tissues and organs of apparently healthy fish implies that piscidin may have other functions in addition to its role as an immune effector molecule.
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Affiliation(s)
- Jareeporn Ruangsri
- Faculty of Biosciences and Aquaculture, University of Nordland, 8049, Bodø, Norway
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Oehlers SH, Flores MV, Hall CJ, Crosier KE, Crosier PS. Retinoic acid suppresses intestinal mucus production and exacerbates experimental enterocolitis. Dis Model Mech 2012; 5:457-67. [PMID: 22563081 PMCID: PMC3380709 DOI: 10.1242/dmm.009365] [Citation(s) in RCA: 65] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/11/2023] Open
Abstract
Exposure to retinoids for the treatment of acne has been linked to the etiology of inflammatory bowel disease (IBD). The intestinal mucus layer is an important structural barrier that is disrupted in IBD. Retinoid-induced alteration of mucus physiology has been postulated as a mechanism linking retinoid treatment to IBD; however, there is little direct evidence for this interaction. The zebrafish larva is an emerging model system for investigating the pathogenesis of IBD. Importantly, this system allows components of the innate immune system, including mucus physiology, to be studied in isolation from the adaptive immune system. This study reports the characterization of a novel zebrafish larval model of IBD-like enterocolitis induced by exposure to dextran sodium sulfate (DSS). The DSS-induced enterocolitis model was found to recapitulate several aspects of the zebrafish trinitrobenzene-sulfonic-acid (TNBS)-induced enterocolitis model, including neutrophilic inflammation that was microbiota-dependent and responsive to pharmacological intervention. Furthermore, the DSS-induced enterocolitis model was found to be a tractable model of stress-induced mucus production and was subsequently used to identify a role for retinoic acid (RA) in suppressing both physiological and pathological intestinal mucin production. Suppression of mucin production by RA increased the susceptibility of zebrafish larvae to enterocolitis when challenged with enterocolitic agents. This study illustrates a direct effect of retinoid administration on intestinal mucus physiology and, subsequently, on the progression of intestinal inflammation.
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Affiliation(s)
- Stefan H Oehlers
- Department of Molecular Medicine and Pathology, School of Medical Sciences, The University of Auckland, Auckland 1142, New Zealand
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Semple F, Dorin JR. β-Defensins: multifunctional modulators of infection, inflammation and more? J Innate Immun 2012; 4:337-48. [PMID: 22441423 DOI: 10.1159/000336619] [Citation(s) in RCA: 251] [Impact Index Per Article: 19.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/21/2011] [Accepted: 01/17/2012] [Indexed: 12/13/2022] Open
Abstract
Defensins comprise one of the largest groups of host defence peptides, present throughout evolution, in fungi and flowering plants as well as in invertebrates and vertebrates. These cysteine-rich, cationic peptides have a common ability to kill a broad range of microorganisms including bacteria, yeast and viruses. As such, they are a strong component of the arsenal that is an organism's innate immunity. It is becoming increasingly clear, however, that antimicrobial action is only one of the numerous roles of these multifunctional peptides. In recent years, the functions of defensins in immunomodulation have been widely investigated, and their involvement in other processes (such as fertility) is becoming evident. This review addresses recent advances in the immunomodulatory activity of β-defensins as well as the involvement of β-defensins in fertility, development, wound healing and cancer.
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Affiliation(s)
- Fiona Semple
- MRC Human Genetics Unit, Medical Research Council Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK
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