Full length articleEffect of Shilajit enriched diet on immunity, antioxidants, and disease resistance in Macrobrachium rosenbergii (de Man) against Aeromonas hydrophila
Introduction
Freshwater prawn culture has been recognized as an eco-friendly alternative for sustainable prawn production. In many countries the freshwater prawn, Macrobrachium rosenbergii (de Man) is an economically important venture because of its high commercial value [1]. It is a highly priced elite food product in both domestic and export market; M. rosenbergii is euryhaline and survives in a wide range of salinities between 0 and 18‰ [2]. In many countries its farming has been expanding significantly during the last decade. However the juveniles and adults of M. rosenbergii suffer high mortalities especially in hatcheries [3], [4], [5] due to several disease outbreaks such as appendage deformity syndrome (ADS) [6] and bacterial pathogens such as Vibrio spp., Aeromonas spp., and Pseudomonas spp., and Lactococcus garviae [7], [8]. Among, these, Aeromonas spp. is considered to be the major threat in the commercial cultivation of M. rosenbergii in Taiwan [9] and Brazil [10] including India [11], [12].
In the prevention and control of prawn diseases large quantities of antibiotics and chemicals are applied; vaccines are another eco-friendly measure. However the traditional measures build up drug-resistance in pathogens; vaccines are pathogen specific and hence novel strategies to control bacterial diseases in aquaculture are needed. The effect of plant products on innate and adaptive immune response in fish and shellfish diseases was reviewed [13]. It is encouraging to note that in Penaeus monodon dietary administration of polysaccharide gel obtained from the fruit-rind of Durio zibethinus could significantly increase the immune response and disease resistance against Vibrio harveyi and white spot syndrome virus (WSSV) [14]. In M. rosenbergii administration of Withania somnifera and Eichhornia crassipes supplemented diets positively enhance the immunity and survival rate against A. hydrophila and L. garvieae [15], [16].
Shilajit is a blackish-brown exudates of variable consistence obtained from the steep rocks of specific mountain regions of the world at altitudes between 0.6 and 5 km [17]. In India it is found in Himalayas region at altitudes between 1 and 5 km on the cave walls embedded in rocks or as rock exudates from Arunachal Pradesh in the East to Kashmir in the West [17], [18]. Though shilajit has similar physical properties and qualitative chemical composition, there is a regional variation in the ratio of individual components. Shilajit/humus consists of humic (80–85% of total organic mass) and non-humic (15–20%) substances [18], organic matter (60–80%), mineral matter (20–40%), and ∼5% trace elements. Shilajit contain more than 85 minerals in ionic form and humic substances (mainly fulvic and humic acid) [19]. It contains 14–20% humidity; 18–20% minerals; 13–17% proteins (with marked- α-amylase activity); 4–4.5% lipids; 3.3–6.5% steroids; 18–20% nitrogen-free compounds; 1.5–2% carbohydrates; and 0.05–0.08% alkaloids, and a number of amino acids [20]. Shilajit comprise 65 organic compounds namely, albumins, coumarins, free fatty acids, organic acids including adipic, succinic, citric, oxalic and tartaric, waxes, resins, polyphenols, essential oils and vitamins like B1, B12, etc. [21], [22]. Shilajit also contain a number of active constituents such as dibenzo-α-pyrones and related metabolites, such as tirucallane triterpenes, small peptides consisting of non-protein amino acids, some phenolic lipids, small tannoids, and FA [21].
In many countries Shilajit has been used for centuries as a traditional medicine [17] in treatment of genitourinary diseases, diabetes, digestive disorders, nervous diseases, tuberculosis, chronic bronchitis, asthma, jaundice, anemia, eczema, bone fractures, osteoporosis [23], [24], [25] kidney stones, edema, spondylitis, hemorrhoids, injured muscles, bone fractures, and diseases such as osteoporosis and other diseases; it is also used as a rejuvenator and an internal antiseptic [17], [26], [27]. Shilajit has potential use such as anti-inflammatory, anti-fungal, anti-ulcerogenic, anxiolytic activity, anti-allergic, analgesic, anti-diabetic, memory enhancer, and an antioxidant [18], [23], [28], [29], [30], [31]. For therapeutic applications it is administered in the form of an aqueous extract to activate phagocytosis and cytokine release by murine peritoneal macrophages [32], stimulate osteoblastic differentiation of mesenchymal stem cells [33], induce the proliferation of lymphocytes in the cortical thymus layer and increase migration of these cells into thymus-dependent zones of the lymph nodes and spleen [34]. In the aqueous extract of Shilajit humus comprise fulvic acid (FA) as the primary organic substance endowed with for many biological and medicinal properties [25], [29] effective in the treatment of disorders including gastritis, diarrhea, stomach ulcers, dysentery, colitis and diabetes mellitus [17], [25] and stimulate neutrophil and lymphocyte immune function [35], [36]. FA has broad spectrum antimicrobial property on a variety of bacteria, including P. gingivalis, F. nucleatum, S. mitis, A. actinomycetemcomitans, E. faecalis, S. mutans and also create a cytotoxic environment for cancer cells [37]. The complement system plays an essential role in innate immunity, contributing to inflammatory responses and the destruction and removal of pathogens. Likewise, the removal of complement by fixation has also been proposed to be a potential therapeutic strategy for treating inflammatory diseases [38]. It is also used in the form of an aqueous extract for therapeutic applications as an immuno stimulant and anabolic food additive [36]. To our knowledge this is the first study on the protective efficacy of dietary supplementation with Shilajit on antioxidant activities, innate immune function, and disease resistance in M. rosenbergii against A. hydrophila.
Section snippets
Diet
The basal diet (control) comprised mackerel meal, dehulled soybean meal, and corn gluten meal as the protein source; wheat flour, α-potato starch, and wheat gluten as carbohydrate and fish oil as lipid source in addition with vitamin and mineral premix (Table 1). The dietary Shilajit (Aravind Annai Herbals, Chennai, India) was incorporated with the basal diet at doses of 2, 4, and 6 g kg−1 by evenly mixing with the basal diet thoroughly. The enriched feeds were dried in a vacuum freeze drier
Hematology
The THC did not significantly increase in the infected prawn when fed with Shilajit-enriched diets on first week except with 2 g kg−1 diet as compared to control. However, THC significantly increased when fed with each enriched diets on weeks 2 and 4 (Fig. 1).
Respiratory bursts
The respiratory bursts activity did not significantly enhance in any Shilajit-enriched diet except with 2 g kg−1 diet on weeks 1 and 2 when compared to control; while it was significantly enhanced with 2 and 4 g kg−1 diets on fourth week (
Discussion
Globally especially in South East Asia, the freshwater giant prawn, M. rosenbergii is considered as a potential candidate species for sustainable aquaculture. However the production of prawns in countries including India has been vulnerable by many factors including diseases leading to significant economic loss in nursery and grow-out ponds. Among the bacterial pathogens, motile Aeromonas cause high mortality in prawn culture. In aquaculture, application of traditional medicine is
Acknowledgments
The authors express their sincere gratitude to the Dean of Scientific Research, the King Saud University, Riyadh, Saudi Arabia for funding this Research Group project no. RGP-1437 - 005.
References (76)
- et al.
The effect of temperature and salinity on the metabolic rate of juvenile Macrobrachium rosenbergii (Crustacea: Palaemonidae)
Comp. Biochem. Physiol.
(1977) - et al.
Immunostimulation of tiger shrimp (Penaeus monodon) haemocytes for generation of microbiocidal substances: analysis of reactive oxygen species
Dev. Comp. Immunol.
(1994) - et al.
Bacterial flora associated with larval rearing of the giant freshwater prawn, Macrobrachium rosenbergii
Aquaculture
(2002) - et al.
Responses of giant freshwater prawn (Macrobrachium rosenbergii) to challenge by two strains of Aeromonas spp
J. Invertebr. Pathol.
(2000) - et al.
Impact of plant products on innate and adaptive immune system of cultured finfish and shellfish
Aquaculture
(2011) - et al.
Studies on the immunomodulatory effect of polysaccharide gel extracted from Durio zibethinus in Penaeus monodon shrimp against Vibrio harveyi and WSSV
Fish. Shellfish Immunol.
(2010) - et al.
Immunomodulatory effect of Withania somnifera supplementation diet in the giant freshwater prawn Macrobrachium rosenbergii (de Man) against Aeromonas hydrophila
Fish. Shellfish Immunol.
(2012) - et al.
Effects of dietary administration of water hyacinth (Eichhornia crassipes) extracts on the immune responses and disease resistance of giant freshwater prawn, Macrobrachium rosenbergii
Fish. Shellfish Immunol.
(2013) - et al.
Thermal analysis of mumiyo, the legendary folk remedy from the Himalaya region
Thermochim. Acta
(2004) - et al.
Antiulcerogenic and antiinflammatory studies with shilajit
J. Ethnopharmacol.
(1990)
Isolation, partial characterisation and immunomodulating activities of polysaccharides from Vernonia kotschyana Sch. Bip. Ex Walp
J. Ethnopharmacol.
Glutathione peroxidase activity in selenium deficient rat liver
Biochem. Biophys. Res. Commun.
Purification and characterization of flavoenzyme glutathione reductase from rat liver
Biol. Chem.
Protective efficacy of azomite enriched diet in Oreochromis mossambicus against Aeromonas hydrophila
Aquaculture
Effect of hypoxia stress on the immune response and resistance to vibriosis of the shrimp Penaeus stylirostris
Fish. Shellfish Immunol.
Dietary-1,3-glucan effectively improves immunity and survival of Penaeus monodon challenged with white spot syndrome virus
Fish. Shellfish Immunol.
Knowledge and research prospects in marine mollusk and crustacean immunology
Aquaculture
The role of reactive oxygen species in the killing of the bacterial fish pathogen, Aeromonas salmonicida by rainbow trout macrophages
Fish. Shellfish Immunol.
Effects of intrinsic and extrinsic factors on the haemocyte profile of the prawn, Macrobrachium rosenbergii
Fish. Shellfish Immunol.
Role of the prophenoloxidase activating system in invertebrate immunity
Curr. Opin. Immunol.
Comparative study on immune response of Fenneropenaeus indicus to Vibrio alginolyticus and white spot syndrome virus
Aquaculture
Dietary supplement of banana (Musa acuminata) peels hot-water extract to enhance the growth, anti-hypothermal stress, immunity and disease resistance of the giant freshwater prawn, Macrobrachium rosenbergii
Fish. Shellfish Immunol.
Comparison of the effects of four anaesthetics on blood biochemical profiles and oxidative stress biomarkers in rainbow trout
Aquaculture
Antioxidant and membrane bound enzymes activity in WSSV- infected Penaeus monodon Fabricius
Aquaculture
Changes in white spot syndrome virus (WSSV) infected Penaeus monodon
Comp. Biochem. Physiol.
The cytosolic manganese superoxide dismutase from the shrimp Litopenaeus vannamei: molecular cloning and expression
Dev. Comp. Immunol.
Identification and cloning of the antioxidant enzyme, glutathione peroxidase, of white shrimp, Litopenaeus vannamei, and its expression following Vibrio alginolyticus infection
Fish. Shellfish Immunol.
Diseases of the freshwater prawn, Macrobrachium rosenbergii
Aquat. Anim. Health Res. Inst. Newsl.
Disease problems of prawns in recirculation systems in U.K
Aquaculture
Is appendage deformity syndrome caused by Macrobrachium rosenbergii nodavirus?
Curr. Sci.
Lactococcus garvieae infection in the giant freshwater prawn Macrobrachium rosenbergii confirmed by polymerase chain reaction and 6S rDNA sequencing
Dis. Aquat. Org.
Dietary administration of bovine lactoferrin influences the immune ability of the giant freshwater prawn Macrobrachium rosenbergii (de Man) and its resistance against Aeromonas hydrophila infection and nitrite stress
Fish. Shellfish Immunol.
Effect of nucleotide on growth, immune responses and resistance of Macrobrachium rosenbergii (De Man) to Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) and Aeromonas hydrophila infection
Aquacult. Int.
Shilajit: a review
Phytother. Res.
Chemistry of Shilajit, an immunomodulatory ayurvedic rasayan
Pure Appl. Chem.
Effects of shilajit on rat brain monamines
Phytother. Res.
Cited by (24)
A review on the application of herbal medicines in the disease control of aquatic animals
2020, AquacultureCitation Excerpt :Additives from Z. officinale promoted immunological indicators, such as skin and mucosa, in Indian major carp (Labeo rohita), and improved resistance against A. hydrophila infection (Sukumaran et al., 2016). Adding Mumiyo to feed increased the activity of phenoloxidase and superoxide dismutase in Macrobrachium rosenbergii, resulting in reduced mortality following A. hydrophila infection (Musthafa et al., 2016). Myrciaria dubia extracts could also significantly improve the immune response of O. niloticus to A. hydrophila, including lysozyme activity, respiratory activity, serum bactericidal activity, total white blood cells, and direct agglutination (Yunis-Aguinaga et al., 2016).
Dietary montmorillonite as growth promoter and immunomodulator in rainbow trout (Oncorhynchus mykiss)
2020, ChemosphereCitation Excerpt :In this regard, it has been proved that application of various immunostimulant classes or traditional medicines such as natural minerals in aquaculture is an environmentally friendly and sustainable alternative for antibiotics (Song et al., 2014; Ringo and Song, 2016; Wang et al., 2017; Ringo et al., 2018). It has been revealed that natural clay minerals (e.g. zeolite, azomite, bentonite, mordenite, montmorillonite and shilajit) not only can apply as promising approach for avoiding overuse of antibiotics, but also they can use as carriers of antibacterial agents (e.g. Cu2+) in aquaculture (Hu et al., 2007; Tan et al., 2014; Kanyılmaz et al., 2015; Musthafa et al., 2016a, 2016b; Jawahar et al., 2016, 2018). Several characteristics such as great capacity for exchanging ions, extensive surface area and consistency of chemical and mechanical composition, high availability and eco-friendly features make clay minerals as a valuable feed additive in aquaculture (Ismadji et al., 2015).
Insight into the diversity of antibiotic resistance genes in the intestinal bacteria of shrimp Penaeus vannamei by culture-dependent and independent approaches
2019, Ecotoxicology and Environmental SafetyProtective efficacy of Shilajit enriched diet on growth performance and immune resistance against Aeromonas hydrophila in Oreochromis mossambicus
2018, Fish and Shellfish ImmunologyCitation Excerpt :It plays an essential role as a rejuvenator and a potential immuno-stimulant. Shilajit is also able to enhance antioxidant properties [10] and eliminates free radicals due to the presence of bioactive constituents, dibenzo- -pyrones, carotenoids, indigoids, metallo-humus substance like fulvic acids and fusims [11–13]. Furthermore, it comprises numerous organic substances and vitamins (like B1 and B12) [14,15].
Kaolin incorporated diet on growth and immune response in Ctenopharyngodon idellus against Aeromonas hydrophila
2018, Fish and Shellfish Immunology