Elsevier

Fish & Shellfish Immunology

Volume 63, April 2017, Pages 87-96
Fish & Shellfish Immunology

Full length article
Response of gut health and microbiota to sulfide exposure in Pacific white shrimp Litopenaeus vannamei

https://doi.org/10.1016/j.fsi.2017.02.008Get rights and content

Highlights

  • Chronic exposure to sub-lethal sulfide could lead to the damage of gut structure.

  • Sub-lethal sulfide exposure led to inflammatory response and decreased the immunity of white shrimp.

  • The microbiota structure in the gut of L. vannamei could be shaped by sulfide exposure.

Abstract

Sulfide is a natural and widely distributed toxicant. It can be commonly found on the interface between water and sediment in the aquatic environment. The Pacific white shrimp Litopenaeus vannamei starts life in the benthic zone soon after the mysis stage, an early stage of post larvae. Therefore, L. vannamei is inevitably affected by exposure to sulfide released from pond sediment. This study explored the toxicant effect of different concentrations of sulfide on the intestinal health and microbiota of Pacific white shrimp by monitoring the change of expression of inflammatory, immune related cytokines, and the structure of the intestinal microbiota. The gut histology, expressions of inflammatory and immune related cytokines (tumor necrosis factor-alpha, C-type lectin 3, myostatin and heat shock transcription factor 1), and the microbiota were determined in L. vannamei after exposure to 0 (control), 425.5 (1/10 LC 50–96 h), and 851 μg/L (1/5 LC 50–96 h) of sulfide for 21 days. With the increase of sulfide concentration, intestinal injury was aggravated and the inflammatory and immune related cytokines generated a range of reactions. The expression of myostatin (MSTN) was significantly down-regulated by the concentration of sulfide exposure. No difference in the expression of heat shock transcription factor 1 (HSF1) was found between the control and shrimp exposed to 425.5 μg/L, but significantly higher HSF1 expression was found in shrimp exposed to 851 μg/L of sulfide. Significantly higher values of tumor necrosis factor-alpha (TNF-α) and C-type lectin 3 (CTL3) were found in the shrimp exposed to 425.5 μg/L of sulfide compared to the control, but a lower value was found in the shrimp exposed to 851 μg/L (P < 0.05). Sulfide also changed the intestinal microbial communities. The abundance of pathogenic bacteria, such as Cyanobacteria, Vibrio and Photobacterium, increased significantly with exposure to the increasing concentration of sulfide. The abundance of some anti-stress bacteria, such as Chlorobi and Fusobacterium, increased. Nitrospirae which can alleviate nitrite toxicity decreased. Microbacterium, Parachlamydia, and Shewanella were all commonly found and down-regulated in both sulfide groups, which is associated with an adaptation to sulfide stimulation. This study indicates that chronic exposure to sub-lethal levels of sulfide could lead to damage of the gut structure, stimulate the response of the inflammatory and immune systems, and shape the structure of the gut microbiota in L. vannamei.

Introduction

The animal gut is a vital organ for food storage, digestion and nutrient absorption [1]. Additionally, the gut plays a role in immune function together with symbiotic bacteria and serves as a biochemical barrier to contain gastric acid and glycoprotein [2]. The integrity and the inflammatory status of the animal gut have been used to evaluate the gut health status of animals [3]. Usually, a normal gut has closely arranged epithelial cells, a clear intercellular space, an inner wall of columnar epithelial tissue, and villi with an integrated structure [4]. The intestinal epithelial cells separate the basement membrane and will disintegrate if the gut wall is damaged [5]. In addition, a functional and stable gut microbiota is important to the host's health, as the gut microbiota can perform beneficial functions to the host such as balancing the immune response, absorbing nutrients, and maintain homeostasis [6]. Previous studies have shown that manipulation of the microbial composition in the gut of farmed fish and crustaceans can have a marked effect on animal health, growth, and survival [7]. Previous studies have shown that the gut microbiota can contribute to the absorption of fatty acids and the formation of lipid droplets in the intestinal epithelium of zebrafish [8]. Additionally, feeding Epinephelus coioides a diet containing Bacillus subtilis E20 improved growth, innate immune response, and disease resistance [9]. It has been proposed that modulating the gut microbiota will be a clean, ecological and prospective technology in dealing with the challenge of shrimp disease [10]. Therefore, the maintenance of gut physiological function in an animal is of significance for the growth and development of aquatic animals.

Many factors, such as water quality, infection with pathogenic bacteria or virus, and diet composition, can affect the gut health of aquatic animals [11], [12], [13], [14]. Among these factors, water quality can directly impact the gut health of aquatic animals [15], as some toxicants are generated from the intensive aquaculture system [16]. Sulfide is one of these toxicants. It has been associated with fish farming and can affect mitochondrial respiration through the inhibition of cytochrome C oxidase [17], [18] and lead to a condition favorable for bacterial proliferation [19]. Previous studies have found that sulfide can potentially threaten the gut health of aquatic animals [20] and shape the composition of the gut microbiota [21]. However, our understanding regarding the toxic effect of sulfide on the gut health of aquatic animals is still limited. There is a need for further systematic investigation.

Sulfide can be accumulated in an aquatic environment when organic material is decomposed under anaerobic conditions. It is commonly found in the interface between the water and sediment in the aquatic environment [22]. Therefore, the species dwelling in the benthic zone on the bottom of a pond are susceptible to sulfide exposure and toxicity. The Pacific white shrimp Litopenaeus vannamei starts benthic life soon after the mysis stage, or the early stage of post larvae. Therefore, L. vannamei is inevitably affected by exposure to sulfide released from the pond sediment. However, our understanding of the effect of sulfide toxicity on L. vannamei is limited to its immune response and its susceptibility to Vibrio alginolyticus under sulfide stress [13].

In this study, we aimed to examine the impact of sulfide on L. vannamei from the perspectives of gut histology, expression of inflammatory and immune related factors (TNF-α, CTL3, MSTN and HSF1), and the microbiota under chronic, sub-lethal concentrations of sulfide. The lethal concentration (LC50) of sulfide for L. vannamei was pre-determined for 96 h, which provided the basis for choosing the concentration of sulfide used in the present study. The results of this study will provide insight into our understanding of the mechanism of sulfide toxicity for L. vannamei.

Section snippets

Experimental shrimp

Juvenile Pacific white shrimp L. vannamei (3.2 ± 0.16 g) were obtained from South China Sea Fisheries Research Institute in Shenzhen, China. L. vannamei were acclimated at a salinity of 25 psu for one week prior to the start of the experiment. Seawater together with the tap water from Dayawan, Shenzhen was filtered through an activated carbon cartridge for at least three days before use. During the experiment, the water quality parameters in the experiment were maintained at a temperature of

Gut histology

The epithelial cells of the shrimp intestine in the control were arranged closely with a clear cell gap, and the intestinal walls were found to be composed of tissues with a tall, columnar epithelium (Fig. 1-a). There were spaces between the partial intestinal and epithelial cells in shrimp exposed to low sulfide at 425.5 μg/L, and part of the intestinal epithelial cells were disassociated from the basement membrane (Fig. 1-b). The intestinal epithelial cells of shrimp exposed to the high

Discussion

Field investigations have established that sulfide is often present in natural aquatic ecosystems at levels that are toxic to fish and invertebrates [11]. In this study, sulfide exposure led to the injury of the L. vannamei gut, and this intestinal damage intensified with increasing sulfide concentration. The damaged gut in L. vannamei can be related to the toxic mechanism of sulfide to animals. Sulfide inhibits the activity of cytochrome C oxidase, causing damage to aerobic respiration and

Acknowledgements

This study was supported by grants from the Agriculture Ministry Key Laboratory of Healthy Freshwater Aquaculture, the Key Laboratory of Freshwater Aquaculture Genetic and Breeding of Zhejiang Province of the Zhejiang Institute of Freshwater Fisheries (ZJK201503), and the National Natural Science Foundation of China (No. 31472291).

References (68)

  • M. Li et al.

    Identification of a C-type lectin with antiviral and antibacterial activity from pacific white shrimp Litopenaeus vannamei

    Dev. Comp. Immunol.

    (2014)
  • M. Zhang et al.

    Characterization of the intestinal microbiota in Pacific white shrimp, Litopenaeus vannamei, fed diets with different lipid sources

    Aquaculture

    (2014)
  • M. Zhang et al.

    Response of gut microbiota to salinity change in two euryhaline aquatic animals with reverse salinity preference

    Aquaculture

    (2016)
  • M. Tedengren et al.

    Heat pretreatment increases cadmium resistance and HSP 70 levels in Baltic Sea mussels

    Aquat. Toxicol.

    (2000)
  • D. Xu et al.

    Molecular cloning of hsf1 and hsbp1 cDNAs, and the expression of hsf1, hsbp1 and hsp70 under heat stress in the sea cucumber Apostichopus japonicus

    Compa. Biochem. Phys. A

    (2016)
  • M. Zhang et al.

    Response of gut microbiota to salinity change in two euryhaline aquatic animals with reverse salinity preference

    Aquaculture

    (2016)
  • D.S. Dabadé et al.

    Bacterial concentration and diversity in fresh tropical shrimps (Penaeus notialis) and the surrounding brackish waters and sediment

    Int. J. Food Microbiol.

    (2016)
  • H.-S. Kang et al.

    Minutissamides E–L, antiproliferative cyclic lipodecapeptides from the cultured freshwater cyanobacterium cf. Anabaena sp

    Bioorgan Med. Chem.

    (2012)
  • S.Y. Cheng et al.

    Effect of sulfide on the immune response and susceptibility to Vibrio alginolyticus in the kuruma shrimp Marsupenaeus japonicus

    Fish. Shellfish Immunol.

    (2007)
  • S.W. Hsu et al.

    The immune response of white shrimp Penaeus vannamei and its susceptibility to Vibrio alginolyticus under sulfide stress

    Aquaculture

    (2007)
  • F. Liu et al.

    Characterization of two pathogenic Photobacterium strains isolated from Exopalaemon carinicauda causing mortality of shrimp

    Aquaculture

    (2016)
  • J.T. Tzuc et al.
    (2014)
  • Y. Chen et al.

    Digestive tract barrier and the immune function of dendritic cells: research progress

    Chin. J. Microecology

    (2013)
  • S.C. Bischoff

    “Gut health”: a new objective in medicine?

    BMC Med.

    (2011)
  • Q. Li

    The Study on Intestinal Histology and Developing Regulation of Nutrients of Alogynogenetic Crucian

    (2007)
  • X. Qing et al.

    Pathological changes and transcriptional response to immersion infection by Vibrio harveyi in shrimp (Litopenaeus vannamei) gut

    J. Fish. China

    (2015)
  • C. Almansa et al.

    Intestinal microbiota, pathophysiology and translation to probiotic use in patients with irritable bowel syndrome

    Expert Rev. gastroent. Hepatol.

    (2012)
  • D.L. Merrifield et al.

    Aquaculture Nutrition: Gut Health, Probiotics and Prebiotics

    (2014)
  • I. Semova et al.

    Microbiota regulate intestinal absorption and metabolism of fatty acids in the zebrafish

    Cell Host Microbe

    (2011)
  • J. Xiong et al.

    Advances, challenges, and directions in shrimp disease control: the guidelines from an ecological perspective

    Appl. Microbiol Bio

    (2016)
  • R. Greenway et al.

    Patterns of Macroinvertebrate and fish diversity in freshwater sulphide springs

    Diversity

    (2014)
  • M. Konishi et al.

    Effects of hydrogen sulfide on bacterial communities on the surface of galatheid crab, Shinkaia crosnieri, and in a bacterial mat cultured in rearing tanks

    Microbes Environ.

    (2013)
  • M.P. Bryant et al.

    Growth of desulfovibrio in lactate or ethanol media low in sulfate in association with H2-utilizing methanogenic bacteria

    Appl. Environ. Microb.

    (1977)
  • M.M. Dorgham

    Effects of Eutrophication, Eutrophication: Causes, Consequences and Control

    (2014)
  • Cited by (124)

    • Impaired intestinal immunity and microbial diversity in common carp exposed to cadmium

      2024, Comparative Biochemistry and Physiology Part - C: Toxicology and Pharmacology
    View all citing articles on Scopus
    View full text