CC BY
25
RUDN Journal of Agronomy and Animal Industries
Вестник РУДН. Серия: АГРОНОМИЯ И ЖИВОТНОВОДСТВО
2019; 14(3):266—278
http://agrojournal.rudn.ru
Research article
DOI 10.22363/2312-797X-2019-14-3-266-278
Galactooligosaccharide effects as prebiotic on intestinal microbiota of different fish species
Seyed H. Hoseinifar1,2*, Hien V. Doan2, Ghasem Ashouri1
1Polytechnic University of Marche, Ancona, Italy 2Chiang Mai University, Chiang Mai, Thailand * Corresponding author: hossein.hoseinifar@gmail.com
Abstract. Manipulation of the gut microbiota toward potentially beneficial bacteria (probiotics) has beneficial effects on fish physiology and health. The effects of prebiotics on gut microbiota are species specific. The present study aimed at investigation of the effects of galactooligosaccharide (GOS) as prebi-otic on intestinal microbiota of Caspian roach and Caspian white fish fingerlings. which are among the most economically valuable species in the Caspian Sea. The study was conducted in a completely randomized design with two set of experiment each of them include three treatments in triplicates in which 0 (control), 1 and 2% GOS were used in diet for 6 weeks. At the end of the period, changes in the intestinal microbiota, including total bacterial count, lactic acid count and lactic acid bacteria (LAB) levels and dominance of LAB in the intestinal microbiota, were measured by culture-based method. Dietary GOS had no significant effect on total bacterial count in both species (P < 0.05). The LAB levels in the intestinal microbiota in the treatments fed with prebiotics was significantly higher than the control group (P < 0.05). LAB bacteria showed the highest increase and dominance in treatments fed with 2% GOS. Also, the highest ratio of lactic acid bacteria to the total number of viable bacteria was observed in the treatment with 2% GOS treatment (P < 0.05). The results of this study indicated the possibility of alterations in the bacterial communities of Caspian roach and Caspian white fish fingerlings gut toward beneficial bacterial communities using GOS as prebiotic.
Key words: prebiotic, Caspian white fish, Caspian roach, galactooligosaccharide, gut microbiota ACKNOWLEDGMENTS
The author wishes to thank Friesland Foods Domo managers (Zwolle, The Netherlands) which kindly provide prebiotic. In addition, we would like to thank the staff of Ghareshsoo research station for their help during the experiment.
Article history:
Received: 15 July 2019. Accepted: 22 August 2019
For citation:
Hoseinifar SH, Doan HV, Ashouri G. Galactooligosaccharide Effects as prebiotic on intestinal microbiota of different fish species. RUDN Journal of Agronomy and Animal Industries, 2019; 14(3):266—278. doi: 10.22363/2312-797X-2019-14-3-266-278
© Hoseinifar S.H., Doan H.V., Ashouri G., 2019.
This work is licensed under a Creative Commons Attribution 4.0 International License https://creativecommons.org/licenses/by/4.0/1
©
Introduction
The study of gut microbiota is of high importance not only regarding disease but also regarding the status of fish physiology and immunity [1]. Establishment of a healthy microbiome in intestine has direct immune-physiological effects on host. It is now well-demonstrated that there is a direct cross talk between gut microbiota and immune response of fish [2, 3]. With the identification of lactic acid bacteria (LAB) in the intestinal microbiota of different fish and shrimp shellfish species in the last decade and determination of their role in the health, welfare and growth performance of the host, the importance of this group of bacteria has become increasingly clear [4, 5]. Although the presence of LAB in the intestinal microbiota of many fish, including Atlantic cod (Gadus morhua), rainbow trout (Oncorhynchus mykiss), Beluga (Huso huso), Persian sturgeon (Acipenserpersicus) and Arctic charr (Salvelinus alpinus) has been proven, they are not the dominant microbiota and constitute a very limited portion of gut microbiota of these species [6]. In addition, it was not possible to isolate LAB bacteria from several fish species [5, 6]. Given this fact, it has been attempted to increase the number of these bacteria through dietary approaches [7, 8]. One of the most important compounds suggested in this regard are prebiotics, which are compounds that are not absorbed by host organism and consumed by potentially beneficial intestinal bacteria (such as LAB) and increase their numbers [9, 10].
Despite recent studies on the effects of prebiotics on fish growth, immunity and physiological indices, many aspects of their potential for alteration of gut microbiota in aquatic and increasing dominance of beneficial bacteria remained unknown [11]. The previous studies revealed that different prebiotics had different effects on LAB levels and also a single prebiotic had different effects on different fish species. Even in some cases, using high levels of more complex prebiotics (higher degree of polymerization) resulted in adverse effects on total bacterial counts and LAB levels [12]. The contradictory of a prebiotic on different host can be due to difference in intestinal microbiota, physiological condition of digestive tract, etc. [13]. Therefore, determination of a prebiotic effect on intestinal microbiota of different species based on comparative studies will help to identify the best prebiotic to change the gut microbiota for that species.
Galactooligosaccharide (GOS) is one of the most promising prebiotic which previous studies revealed that it could exerts positive effects in different fish species [14—16]. In spite of extensive researches on administration of GOS in fish [17—21], to the best of our knowledge there was no published study the effects of GOS on gut microbiota of different fish species using comparative study. Therefore, in the present study we decided to determine the possible effects of GOS on intestinal microbiota of Caspian roach and Caspian white fish.
Materials and methods
Experimental diets
A commercial feed (Dansu, Iran) was used as a control diet (non-supplemented diet). To prepare experimental diet the basal diet was supplemented with two levels of GOS as prebiotic (1 and 2 %). The ingredients were blended thoroughly in a mixer. Then, water was added and made into pellets. The pellets were air-dried, ground and
sieved to produce a suitable crumble (ca. 500p,m). The experimental diets were stored in plastic bags at -2 °C for further use.
Fish husbandry
The present study was conducted at the Gharasu Fisheries Research Station. The Caspian white (Rutilus kutum) fish and Caspian roach (Rutilus caspicus) fingerlings were supplied by Sijowal Caspian Sea Teleost Fish Propagation & Cultivation Centre (Golestan province, Iran). Fish with mean weight of 1.3 g were stocked in nine separate tanks for each species (totally 18 tanks) at density of 30 fish per tank. The fish were acclimated to lab condition for 2 weeks and then feeding with experimental diets were started. During acclimation, fish were fed with control diet. The culture system water was closed with constant aeration. To maintain water quality every 2 days 50 % of water was exchanged. The water quality parameters were controlled and maintained at optimum levels.
Prebiotic
The prebiotic used in the presents study was GOS that was kindly supplied by Friesland Foods Domo Company (Zwolle, The Netherlands). The commercial product name was Vivinal-GOS® and obtained through the enzymatic conversion of lactose and mainly consists of galactose and glucose molecules.
Evaluation of gut microbiota
Total viable autochthonous heterotrophic aerobic bacteria and LAB levels were determined at the start of trial from 15 specimens from the initial pool of fish. Also, at the end of the feeding trial (week 8) microbiological studies were performed. Fish were starved for 24 h to study the autochthonous microbiota. Three specimen were randomly selected from each tank (i.e. n = 9 per treatment).The intestine of fish were assessed and prepared for bacteria culture as we described on previous study [12]. Briefly, the surface bacteria were killed before dissection using 0.1 % benzalkonium chloride. With utmost care to be aseptic, the intestine of samples obtained, washed with sterile saline and homogenized using tissue homogenizer (Potter-Elvehjem, USA). The homogenized intestine was serially diluted to 10-7 by using sterile saline (0.85 % NaCl). Then, to dermine the level of total bacteria and LAB a portion of the diluates (100 ^L) was spread onto plate count agar (PCA) (Merck, Germany) and de Man, Rogosa and Sharpe (MRS) agar (Merck, Germany), respectively. The seeded plates were incubated at room temperature (25°C) for 5 days [22]. Thereafter, the colony forming units (CFU) g-1 were counted from statistically viable plates (i.e. plates containing 30-300 colonies)[23].
Statistical analysis
Prior to statistical analysis, the normality of data and homogeneity of variance were checked and confirmed. Then, the statistically significant difference (at P < 0.05) between treatments was checked using One-way analysis of variance (ANOVA) followed by Duncan's multiple range tests (36). All statistical analysis were performed using SPSS 16.0 (SPSS Inc., Chicago, IL, USA). The figures were drawn using Excel software (Micorsoft Office ver. 2016).
Results and discussion
The total viable autochthonous heterotrophic aerobic bacteria (THAB) level (Log CFU/g) in the intestine of Caspian roach and Caspian white fish fingerlings fed with different levels of galactaligosaccharide (GOS) as prebiotics is shown in Figure 1. At the beginning of the feeding trial, the THAB of intestinal microbiota was 5.10 ± 0.24 log CFU/g. As shown in Figure 1 A, dietary administration of 1 or 2% GOS in diet had no significant effect on THAB counts in the gut microbiota of Caspian roach (P > 0.05). Similar result was noticed in case of the gut microbiota of Caspian white fish (P > 0.05) (Figure 1 B).
9 8 7 6
'co 5 O 4
M
° 3 J 3
2 1 0
A
a
a
Control
1% GOS
2% GOS
9 8 7 6
'co 5
|Jh 4
O 4
M
° 3 J 3
2 1 0
B
a
Control
1% GOS
2% GOS
a
a
Fig. 1. The effects of different levels of galactoaligosaccharide (GOS) as prebiotic on total bacterial counts (log CFU/g) in Caspian roach (A) and Caspian white fish (B) fingerlings. The bars (mean ±SD) assigned similar letters indicate no significant difference (P > 0.05)
The effects of different levels of GOS prebiotics on the level of LAB of lactic acid bacteria (Log CFU/g) in the gut microbiota of Caspian roach and Caspian white fish fingerling sare summarized in Figure 2. At the beginning of the period, no lactic acid bacteria were isolated from the gut microbiota of both fish species. Indeed, the number of LAB in the intestinal microbiota were statistically too few to count (TFTC; lower than 30 colonies in the first dilution). Similarly, at the end of trial in case of both fish species the LAB levels were TFTC in the control treatment. While, feeding with GOS caused significant increase of LAB level in gut microbiota of the Caspian roach and Caspian white fish fingerlings. In both species, the highest LAB level was noticed in gut microbiota of fish fed with 2% GOS. There were significant difference between 1% GOS and 2% GOS treatment in case of gut microbiota LAB level in Caspian roach (P < 0.05). However, no significant difference was noticed in case of 1 and 2% GOS in Caspian white fish (P > 0.05).
5 4,5 4 3,5
тЧ '
WI 3 &
to 2,5 U 6/j 2 о z
J 1,5 1 0,5 0
B
Control
1% GOS
2% GOS
Fig. 2. The effects of different levels of galactoaligosaccharide (GOS) as prebiotic on Lactic acid bacteria levels (log CFU/g) in Caspian roach (A) and Caspian white fish (B) fingerlings. The bars (mean ±SD) assigned with different letters indicate significant difference (P < 0.05)
a
b
In addition, we calculated the ratio of LAB (potentially useful probiotic bacteria) to THAB in the gut microbiota of both species to see the alteration in the dominance of LAB in gut microbiota (Table 1). The obtained results showed that the ratio of LAB to THAB in all prebiotic treatments was significantly higher than the control treatment (P < 0.05). The highest increase in the ratio of lactic acid bacteria to the total number of viable bacteria was observed in the 2% GOS treatment (P < 0.05). Although the addition of GOS to Caspian white fish diet significantly increased the ratio of lactic acid bacteria, this increment was not dose dependent; there was no significant difference between 1 and 2 % levels (P < 0.05).
Table 1
The ratio (%) of lactic acid bacteria to the total viable bacteria in the gut microbiota of Caspian roach and Caspian white fish fingerlings fed with different levels of GOS as prebiotic. The data in a row (mean ± SD) assigned with different letters indicate significant difference (P< 0.05)
Fish species Treatments
Control 1% GOS 2% GOS
Caspian roach TFTCc 1.70±0.34b 4.84 ± 0.81a
Caspian white fish TFTCb 4.45±0.16a 4.12 ± 0.41a
The intestinal microbiota of the fish includes a complex and diverse community of aerobic and anaerobic bacteria. One of the group of bacteria in the gut microbiota are LAB that are of great importance nowadays as probiotics [24]. Although isolation of lactic acid bacteria from the gut microbiota of various species of fish has been reported, these bacteria are not among the predominant bacterial communities in the gut and are present in low abundance [5]. Lactic acid bacteria are capable of inhibiting the growth of pathogenic bacteria through excretion of bacteriocins and thereby can pose positive effects on the health status and disease resistance of fish [25]. Although identification of the gut microbiota of fish and its manipulating is complex and is not fully understood, providing knowledge regarding possible alternative for modulation of gut microbiota toward beneficial populations is of high importance and can be a promising strategy for enhancing immunity and disease resistance [26—28]. This strategy can help to reduce utilization of antibiotics in aquaculture which per results in sustainable aquaculture [11]. One of the proposed methods for modulation of the intestinal microbiota composition is the use of dietary supplements such as prebiotics [2, 7—9, 29]. To date, many studies have been conducted on the beneficial effects of prebiotics on humans and pets, and in recent years, the use of these supplements in the diet of fish and other aquatic animals has been considered. The efficacy and efficacy of prebiotics have been shown to be influenced by the degree of polymerization, fermentabulity, host species, resident gut microbiota [8]. Therefore, considering the inter-species variation, in order to ensure the beneficial effect of the prebiotic used in the diet and the optimal prebiotic selection, comparative studies should be done.
The results of the present study revealed no significant alteration in THAB in the gut microbiota of both Caspian roach and Caspian white fish (Figure 1). In line with the findings of the present study, dietary administration of inactive yeast (Saccharomyces cerevisiae)
and prebiotic fructoaligosaccharide (FOS) had no significant effects on THAB in the intestinal microbiota of Beluga sturgeon (Huso huso) [12, 30]. Similarly, feeding turbot with FOS supplemented diet exerts no significant effect on THAB [31]. On the other hand, negative effects on THAB level was reported in beluga fed with inulin [32]. The inability of dietary prebiotic to alter the THAB seems to be due to the limited binding sites in the gut [1, 33]. Indeed, the previous studies revealed that prebiotics seems to change the balance of gut microbiota by providing energy source for the beneficial bacteria rather than increasing the THAB. Therefore, the THAB cannot be altered very much due to the limited binding sites.
Concerning the effects of the tested prebiotic (GOS) on potentially useful intestinal bacteria, the results indicated a significant increase in the number of LAB in the intestinal microbiota of both Caspian white fish and roach compared to the control treatment. The highest increase was observed in fish fed with 2% GOS. Previous studies on the aquatic gut microbiota revealed despite the limited number of LAB in the gut microbiota, these potentially useful (probiotic) bacteria can be increased through administration of optimum prebiotics and become dominant bacterial communities [8]. Although there is no comparative study regarding the effects of prebiotics on the composition of the gut microbiota of Caspian white fish and roach, the results of this study are consistent with those of Hoseinifar, Mirvaghefi, Amoozegar, Merrifield, Ring0 [34] that showed the use of GOS (as a prebiotic) in the diet significantly increased the number of LAB in the gut microbiota of rainbow trout. In addition, the use of FOS and yeast prebiotics significantly increased the number of LAB in the gut microbiota of Beluga [12, 30]. Similar results have been observed regarding the effects of FOS on the levels of probiotic bacteria in the intestinal microbiota of Turbot [31]. However, in contrast with these finding, inulin had no significant effect on LAB levels in the intestinal microbiota of the Beluga [35]. Despite the several reports on the prebiotic effects of GOS on physiological and health indices of fish, there are limited reports on the prebiotic effect on the intestinal microbiota composition of fish. According to the results of the present study, GOS is an effective prebiotic for modulation of gut microbiota of both species. The observed differences regarding the dose can be due to differences in the physiological characteristics of the gut, the prebiotic type and the microbiota composition of the gut of these species.
In conclusion, the results of this study showed that the use of GOS can be taken into account as an effective prebiotic in Caspian roach and Caspian white fish diet, aimed at modulation of the balance of gut microbiota toward beneficial bacteria. However, determining the possible effects on physiological parameters as well as mode of action needs further investigation.
References
1. Llewellyn MS, Boutin S, Hoseinifar SH, Derome N. Teleost microbiomes: the state of the art in their characterization, manipulation and importance in aquaculture and fisheries. Frontiers in Microbiology. 2014; 5:207. doi: 10.3389/fmicb.2014.00207
2. Nawaz A, Bakhsh javaid A, Irshad S, Hoseinifar SH, Xiong H. The functionality of prebiotics as immunostimulant: Evidences from trials on terrestrial and aquatic animals. Fish & Shellfish Immunology. 2018; 76:272—278. doi: 10.1016/j.fsi.2018.03.004
3. Lazado CC, Caipang CMA. Mucosal immunity and probiotics in fish. Fish & Shellfish Immunology. 2014; 39(1):78—89. doi: 10.1016/j.fsi.2014.04.015
4. Van Doan H, Hoseinifar SH, Ringo E, Angeles Esteban M, Dadar M, Dawood MA, et al. Host-associated probiotics: A key factor in sustainable aquaculture. Reviews in Fisheries Science & Aquaculture. 2019; 1—27. doi: 10.1080/23308249.2019.1643288
5. Ringo E, Hoseinifar SH, Ghosh K, Doan HV, Beck BR, Song SK. Lactic Acid bacteria in finfish—an update. Frontiers in Microbiology. 2018; 9:1818. doi: 10.3389/fmicb.2018.01818
6. Merrifield DL, Balcazar JL, Daniels C, Zhou Z, Carnevali O, Sun YZ et al. Indigenous lactic acid bacteria in fish and crustaceans. In: Merrifield DL, Ringo E. (eds.) Aquaculture Nutrition: gut health, probiotics and prebiotics. Oxford: John Wiley & Sons; 2014. p.128—168.
7. Doan HV, Hoseinifar SH, Esteban MÂ, Dadar M, Thu TTN. Mushrooms, seaweed, and their derivatives as functional feed additives for aquaculture: an updated view. Studies in Natural Products Chemistry. 2019; 62:41—90. doi: 10.1016/B978-0-444-64185-4.00002-2
8. Ringo E, Zhou Z, Vecino JLG, Wadsworth S, Romero J, Krogdahl A, et al. Effect of dietary components on the gut microbiota of aquatic animals. A never-ending story? Aquaculture Nutrition. 2016; 22(2):219—282. doi: 10.1111/anu.12346
9. Dawood MAO, Koshio S. Recent advances in the role of probiotics and prebiotics in carp aquaculture: A review. Aquaculture. 2016; 454:243—251. doi: 10.1016/j.aquaculture.2015.12.033
10. 1Hoseinifar SH, Esteban MÂ, Cuesta A, Sun Y-Z. Prebiotics and fish immune response: A review of current knowledge and future perspectives. Reviews in Fisheries Science & Aquaculture. 2015; 23(4):315—328. doi: 10.1080/23308249.2015.1052365
11. Lieke T, Meinelt T, Hoseinifar SH, Pan B, Straus DL, Steinberg CEW. Sustainable aquaculture requires environmental-friendly treatment strategies for fish diseases. Reviews in Aquaculture. 2019. doi: 10.1111/raq.12365
12. Hoseinifar SH, Mirvaghefi A, Mojazi Amiri B, Rostami HK, Merrifield DL. The effects of oligofructose on growth performance, survival and autochthonous intestinal microbiota of beluga (Huso huso) juveniles. Aquaculture Nutrition. 2011; 17(5):498—504. doi: 10.1111/j.1365-2095.2010.00828.x
13. Daniels C, Hoseinifar SH. Prebiotic Applications in shellfish. In: Merrifield LD, Ringo E (eds.) Aquaculture Nutrition: gut health, probiotics and prebiotics. Oxford: John Wiley & Sons; 2014. p.401—418.
14. Zhou Q-C, Buentello JA, Gatlin Iii DM. Effects of dietary prebiotics on growth performance, immune response and intestinal morphology of red drum (Sciaenops ocellatus). Aquaculture. 2010; 309(1—4):253—257. doi: 10.1016/j.aquaculture.2010.09.003
15. Burr G, Hume M, Ricke S, Nisbet D, Gatlin D. In Vitro and in vivo evaluation of the prebiotics GroBiotic®-A, inulin, mannanoligosaccharide, and galactooligosaccharide on the digestive microbiota and performance of hybrid striped bass (Morone chrysops x Morone saxatilis). Microbial ecology. 2010; 59(1):187—198. doi: 10.1007/s00248-009-9597-6
16. Grisdale-Helland B, Helland SJ, Gatlin III DM. The effects of dietary supplementation with mannanoligosaccharide, fructooligosaccharide or galactooligosaccharide on the growth and feed utilization of Atlantic salmon (Salmo salar). Aquaculture. 2008; 283(1—4):163—167. doi: 10.1016/j.aquaculture.2008.07.012
17. Yousefi S, Hoseinifar SH, Paknejad H, Hajimoradloo A. The effects of dietary supplement of galactooligosaccharide on innate immunity, immune related genes expression and growth performance in zebrafish (Danio rerio). Fish & Shellfish Immunology. 2018; 73:192—196. doi: 10.1016/j.fsi.2017.12.022
18. Hoseinifar SH, Ahmadi A, Khalili M, Raeisi M, Van Doan H, Caipang CM. The study of antioxidant enzymes and immune-related genes expression in common carp (Cyprinus carpio) fingerlings fed different prebiotics. Aquaculture Research. 2017; 48(11):5447—5454. doi: 10.1111/are.13359
19. Modanloo M, Soltanian S, Akhlaghi M, Hoseinifar SH. The effects of single or combined administration of galactooligosaccharide and Pediococcus acidilactici on cutaneous mucus immune parameters, humoral immune responses and immune related genes expression in
common carp (Cyprinus carpio) fingerlings. Fish & Shellfish Immunology. 2017; 70:391— 397. doi: 10.1016/j.fsi.2017.09.032
20. Guerreiro I, Couto A, Machado M, Castro C, Pousäo-Ferreira P, Oliva-Teles A, et al. Prebiotics effect on immune and hepatic oxidative status and gut morphology of white sea bream (Diplodus sargus). Fish & Shellfish Immunology. 2016; 50:168—174. doi: 10.1016/j.fsi.2016.01.023
21. Miandare HK, Farvardin S, Shabani A, Hoseinifar SH, Ramezanpour SS. The effects of galactooligosaccharide on systemic and mucosal immune response, growth performance and appetite related gene transcript in goldfish (Carassius auratus gibelio). Fish & Shellfish Immunology. 2016; 55:479—483. doi: 10.1016/j.fsi.2016.06.020
22. Hoseinifar SH, Khalili M, Sun YZ. Intestinal histomorphology, autochthonous microbiota and growth performance of the oscar (Astronotus ocellatus Agassiz, 1831) following dietary administration of xylooligosaccharide. Journal of Applied Ichthyology. 2016; 32(6):1137— 1141. doi: 10.1111/jai.13118
23. Rawling M, Merrifield D, Kühlwein H, Snellgrove D, Gioacchini G, Carnevali O, et al. Dietary modulation of immune response and related gene expression profiles in mirror carp (Cyprinus carpio) using selected exotic feed ingredients. Aquaculture. 2014; 418:177—184. doi: 10.1016/j. aquaculture .2013.10.002
24. Van Doan H, Hoseinifar SH, Ringo E, Angeles Esteban M, Dadar M, Dawood MA, et al. Host-associated probiotics: a key factor in sustainable aquaculture. Reviews in Fisheries Science & Aquaculture. 2019; 1—27. doi: 10.1080/23308249.2019.1643288
25. Hoseinifar SH, Sun Y, Wang A, Zhou Z. Probiotics as means of diseases control in aquaculture, A Review of current knowledge and future perspectives. Frontiers in Microbiology. 2018; 9:2429. doi: 10.3389/fmicb.2018.02429
26. Van Doan H, Hoseinifar SH, Tapingkae W, Seel-audom M, Jaturasitha S, Dawood MA, et al. Boosted growth performance, mucosal and serum immunity, and disease resistance nile tilapia (Oreochromis niloticus) fingerlings using corncob-derived xylooligosaccharide and lactobacillus plantarum CR1T5. Probiotics and antimicrobial proteins. 2019; 1—12. doi: 10.1007/s12602-019-09554-5
27. Dawood MAO, Koshio S, Esteban MA. Beneficial roles of feed additives as immunostimulants in aquaculture: a review. Reviews in Aquaculture. 2018; 10(4):950—974. doi: 10.1111/raq.12209
28. Abdel-Tawwab M, Abdel-Rahman AM, Ismael NEM. Evaluation of commercial live bakers' yeast, Saccharomyces cerevisiae as a growth and immunity promoter for Fry Nile tilapia, Oreochromis niloticus (L.) challenged in situ with Aeromonas hydrophila. Aquaculture. 2008; 280(1—4):185—189. doi: 10.1016/j.aquaculture.2008.03.055
29. Ringo E, Dimitroglou A, Hoseinifar SH, Davies SJ. Prebiotics in Finfish: an update. In: Merrifield DL, Ringo E. (eds.) Aquaculture nutrition: gut health, probiotics and prebiotics. Oxford: John Wiley & Sons; 2014. p.360—400. doi: 10.1002/9781118897263.ch14
30. Hoseinifar SH, Mirvaghefi A, Merrifield DL. The effects of dietary inactive brewer's yeast Saccharomyces cerevisiae var. ellipsoideus on the growth, physiological responses and gut microbiota of juvenile beluga (Huso huso). Aquaculture. 2011; 318(1—2):90—94. doi: 10.1016/j.aquaculture.2011.04.043
31. Mahious AS, Gatesoupe FJ, Hervi M, Metailler R, Ollevier F. Effect of dietary inulin and oligosaccharides as prebiotics for weaning turbot, Psetta maxima (Linnaeus, C. 1758). Aquaculture International. 2006; 14(3):219—229. doi: 10.1007/s10499-005-9003-4
32. Akrami R, Ghelichi A, Manuchehri H. Effect of dietary inulin as prebiotic on growth performance and survival of juvenile rainbow trout (Oncorhynchus mykiss). Journal of marine science and technology research. 2009; 4(3):1—9.
33. Li X, Ringo E, Hoseinifar SH, Lauzon HL, Birkbeck H, Yang D. The adherence and colonization of microorganisms in fish gastrointestinal tract. Reviews in Aquaculture. 2019; 11(3):603—618. doi: 10.1111/raq.12248
34. Hoseinifar SH, Mirvaghefi A, Amoozegar MA, Merrifield D, Ringe E. In vitro selection of a synbiotic and in vivo evaluation on intestinal microbiota, performance and physiological response of rainbow trout (Oncorhynchus mykiss) fingerlings. Aquaculture Nutrition. 2015; 23(1):111—118. doi: 10.1111/anu.12373
35. Reza A, Abdolmajid H, Abbas M, Abdolmohammad AK. Effect of dietary prebiotic inulin on growth performance, intestinal microflora, body composition and hematological parameters of juvenile beluga, Huso huso (Linnaeus, 1758). Journal of the World Aquaculture Society. 2009; 40(6):771—779. Available from: doi: 10.1111/j. 1749-7345.2009.00297.x
About authors:
Hoseinifar Seyed Hossein — Phd, Department of Life and Environmental Sciences, Polytechnic University of Marche, 22 Piazza Roma, Ancona, Italy, 60100; Department of Animal and Aquatic Sciences, Faculty of Agriculture, Chiang Mai University, Su Thep, Mueang Chiang Mai District, Chiang Mai, Thailand, 50200; e-mail: hossein.hoseinifar@gmail.com; ORCID: 0000-0002-0210-9013 Doan Hien Van — Phd, Department of Animal and Aquatic Sciences, Faculty of Agriculture, Chiang Mai University, 239 HuayKaew Road, Muang District, Chiang Mai, Thailand, 50200; WOS - N-9579-2019
Ashouri Ghasem — Department of Life and Environmental Sciences, Polytechnic University of Marche, 22 Piazza Roma, Ancona, Italy, 60100
Научная статья
DOI 10.22363/2312-797X-2019-14-3-266-278
Влияние галактоолигосахаридов в виде пребиотика на микрофлору кишечника различных видов рыб
С.Х. Хосейнифар*1'2, Х.В. Доан2, Г. Ашоури1
1 Политехнический Университет Марке, Италия, Анкона 2 Чиангмайский университет, Чиангмай, Таиланд * hossein.hoseinifar@gmail.com
Аннотация. Обогащение кишечной микробиоты потенциально полезными бактериями (про-биотиками) оказывает благотворное влияние на физиологические процессы и здоровье рыб. Однако, воздействие пребиотиков на микрофлору кишечника является видоспецифичным. Настоящее исследование направлено на изучение влияния галактоолигосахаридов в качестве пребиотика на кишечную микробиоту каспийской плотвы и мальков каспийского кутума, являющихся одними из наиболее экономически ценных видов рыб, обитающих в Каспийском море. Исследование проводилось в течение 6 недель по полной рандомизированной схеме, в двух повторениях, каждое из которых включало три варианта обработки — 0 (контроль), 1 и 2 % ГОС, в трехкратной повторности. После этого с помощью культурального метода были изучены изменения в микробиоте кишечника рыб, включая общее количество бактерий, количество молочной кислоты и молочнокислых бактерий, а также влияние молочнокислых бактерий на микрофлору кишечника. Диетические галактоолигос-ахариды не оказали значительного влияния на общее количество бактерий у обоих видов (P < 0.05). Уровень молочнокислых бактерий в кишечнике был значительно выше при лечении пребиотиками, чем в контрольной группе (P < 0.05). Значительное увеличение количества молочнокислых бактерий и их преобладание было отмечено в варианте с использованием 2 % галактоолигосахаридов. Кроме
того, самое высокое количество молочнокислых бактерий по отношению к общему количеству жизнеспособных бактерий наблюдалось в варианте с использованием 2 % галактоолигосахаридов (P < 0.05). Результаты данного исследования доказывают возможность и эффективность использования галактоолигосахаридов в качестве пребиотика для обогащения кишечной бактериальной микрофлоры каспийской плотвы и мальков каспийского кутума.
Ключевые слова: пребиотик, каспийский кутум, каспийская плотва, галактоолигосахарид, кишечная микробиота
БЛАГОДАРНОСТИ
Авторы выражают благодарность сотрудникам Friesland Foods Domo (Зволле, Нидерланды), любезно предоставившим препараты-пребиотики. Также мы хотели бы поблагодарить сотрудников исследовательской станции Ghareshsoo за помощь в проведении научных экспериментов.
История статьи:
Поступила в редакцию: 15 июля 2019 г. Принята к публикации: 22 августа 2019 г. Для цитирования:
Hoseinifar S.H., Doan H.V., Ashouri G. Galactooligosaccharide effects as prebiotic on intestinal microbiota of different fish species // Вестник Российского университета дружбы народов. Серия: Агрономия и животноводство. 2019. Т. 14. № 3. С. 266—278. doi: 10.22363/2312-797X-2019-14-3-266-278
Библиографический список
1. Llewellyn M.S., Boutin S., Hoseinifar S.H., Derome N. Teleost microbiomes: the state of the art in their characterization, manipulation and importance in aquaculture and fisheries // Frontiers in Microbiology. 2014. № 5. P. 207. doi: 10.3389/fmicb.2014.00207
2. Nawaz A., Bakhsh javaid A., Irshad S., Hoseinifar S.H., Xiong H. The functionality of prebiotics as immunostimulant: Evidences from trials on terrestrial and aquatic animals // Fish & Shellfish Immunology. 2018. № 76. P. 272—278. doi: 10.1016/j.fsi.2018.03.004
3. Lazado C.C., Caipang C.M.A. Mucosal immunity and probiotics in fish // Fish & Shellfish Immunology. 2014. Vol. 39. № 1. P. 78—89. doi: 10.1016/j.fsi.2014.04.015
4. Van Doan H., Hoseinifar S.H., Ringo E., Angeles Esteban M., Dadar M., Dawood M.A., et al. Host-associated probiotics: A key factor in sustainable aquaculture // Reviews in Fisheries Science & Aquaculture. 2019. P. 1—27. doi: 10.1080/23308249.2019.1643288
5. Ringo E., Hoseinifar S.H., Ghosh K., Doan H.V,. Beck B.R., Song S.K. Lactic Acid bacteria in finfish — an update // Frontiers in Microbiology. 2018. № 9. Р. 1818. doi: 10.3389/fmicb.2018.01818
6. Merrifield D.L., Balcâzar J.L., Daniels C., Zhou Z., Carnevali O., Sun Y.Z. et al. Indigenous lactic acid bacteria in fish and crustaceans // Aquaculture Nutrition: gut health, probiotics and prebiotics / Merrifield D.L., Ringo E., eds. Oxford : John Wiley & Sons, 2014. P. 128—168.
7. Doan H.V., Hoseinifar S.H., Esteban M.A., Dadar M., Thu T.T.N. Mushrooms, seaweed, and their derivatives as functional feed additives for aquaculture: an updated view // Studies in Natural Products Chemistry. 2019. № 62. P. 41—90. doi: 10.1016/B978-0-444-64185-4.00002-2
8. Ringo E., Zhou Z., Vecino J.L.G., Wadsworth S., Romero J., Krogdahl A., et al. Effect of dietary components on the gut microbiota of aquatic animals. A never-ending story? // Aquaculture Nutrition. 2016. Vol. 22. № 2. P. 219—282. doi: 10.1111/anu.12346 "
9. DawoodM.A. O., Koshio S. Recent advances in the role of probiotics and prebiotics in carp aquaculture: A review // Aquaculture. 2016. № 454. P. 243—251. doi: 10.1016/ j. aquaculture .2015.12.033
10. Hoseinifar S.H., Esteban M.A., Cuesta A., Sun Y.-Z. Prebiotics and fish immune response: A review of current knowledge and future perspectives // Reviews in Fisheries Science & Aquaculture. 2015. Vol. 23. № 4. P. 315—328. doi: 10.1080/23308249.2015.1052365
11. Lieke T., Meinelt T., Hoseinifar S.H., Pan B., Straus D.L., Steinberg C.E.W. Sustainable aquaculture requires environmental-friendly treatment strategies for fish diseases // Reviews in Aquaculture. 2019. doi: 10.1111/raq.12365
12. Hoseinifar S.H., Mirvaghefi A., Mojazi Amiri B., Rostami H.K., Merrifield D.L. The effects of oligofructose on growth performance, survival and autochthonous intestinal microbiota of beluga (Huso huso) juveniles // Aquaculture Nutrition. 2011. Vol. 17. № 5. P. 498—504. doi: 10.1111/j.1365-2095.2010.00828.x
13. Daniels C., Hoseinifar S.H. Prebiotic Applications in shellfish // Aquaculture Nutrition: gut health, probiotics and prebiotics / Merrifield L.D., Ringo E., eds. Oxford : John Wiley & Sons, 2014. P. 401—418. "
14. Zhou Q.-C., Buentello J.A., Gatlin Iii D.M. Effects of dietary prebiotics on growth performance, immune response and intestinal morphology of red drum (Sciaenops ocellatus). Aquaculture. 2010. Vol. 309. № 1—4. Р. 253—257. doi: 10.1016/j.aquaculture.2010.09.003
15. Burr G., Hume M., Ricke S., Nisbet D., Gatlin D. In Vitro and in vivo evaluation of the prebiotics GroBiotic®-A, inulin, mannanoligosaccharide, and galactooligosaccharide on the digestive microbiota and performance of hybrid striped bass (Morone chrysops x Morone saxatilis) // Microbial ecology. 2010. Vol. 59. № 1. P. 187—198. doi: 10.1007/s00248-009-9597-6
16. Grisdale-Helland B., Helland S.J., Gatlin III D.M. The effects of dietary supplementation with mannanoligosaccharide, fructooligosaccharide or galactooligosaccharide on the growth and feed utilization of Atlantic salmon (Salmo salar) // Aquaculture. 2008. Vol. 283. № 1— 4. Р. 163—167. doi: 10.1016/j.aquaculture.2008.07.012
17. Yousefi S., Hoseinifar S.H., PaknejadH., Hajimoradloo A. The effects of dietary supplement of galactooligosaccharide on innate immunity, immune related genes expression and growth performance in zebrafish (Danio rerio) // Fish & Shellfish Immunology. 2018. № 73. Р. 192—196. doi: 10.1016/j.fsi.2017.12.022
18. Hoseinifar S.H., Ahmadi A., Khalili M., Raeisi M., Van Doan H., Caipang C.M. The study of antioxidant enzymes and immune-related genes expression in common carp (Cyprinus carpio) fingerlings fed different prebiotics // Aquaculture Research. 2017. Vol. 48. № 11. Р. 5447—5454. doi: 10.1111/are.13359
19. Modanloo M., Soltanian S., Akhlaghi M., Hoseinifar S.H. The effects of single or combined administration of galactooligosaccharide and Pediococcus acidilactici on cutaneous mucus immune parameters, humoral immune responses and immune related genes expression in common carp (Cyprinus carpio) fingerlings // Fish & Shellfish Immunology. 2017. № 70. Р. 391—397. doi: 10.1016/j.fsi.2017.09.032
20. Guerreiro I., Couto A., Machado M., Castro C., Pousäo-Ferreira P., Oliva-Teles A., et al. Prebiotics effect on immune and hepatic oxidative status and gut morphology of white sea bream (Diplodus sargus) // Fish & Shellfish Immunology. 2016. № 50. Р. 168—174. doi: 10.1016/j.fsi.2016.01.023
21. Miandare H.K., Farvardin S., Shabani A., Hoseinifar S.H., Ramezanpour S.S. The effects of galactooligosaccharide on systemic and mucosal immune response, growth performance and appetite related gene transcript in goldfish (Carassius auratus gibelio) // Fish & Shellfish Immunology. 2016. № 55. Р. 479—483. doi: 10.1016/j.fsi.2016.06.020
22. Hoseinifar S.H., Khalili M., Sun Y.Z. Intestinal histomorphology, autochthonous microbiota and growth performance of the oscar (Astronotus ocellatus Agassiz, 1831) following dietary administration of xylooligosaccharide // Journal of Applied Ichthyology. 2016. Vol. 32 № 6. Р. 1137—1141. doi: 10.1111/jai.13118
23. RawlingM., MerrifieldD., Kühlwein H., Snellgrove D., Gioacchini G., Carnevali O., et al. Dietary modulation of immune response and related gene expression profiles in mirror carp (Cyprinus carpio) using selected exotic feed ingredients // Aquaculture. 2014. № 418. Р. 177—184. doi: 10.1016/j.aquaculture.2013.10.002
24. Van Doan H., Hoseinifar S.H., Ringo E., Angeles Esteban M., Dadar M., Dawood M.A., et al. Host-associated probiotics: a key factor in sustainable aquaculture // Reviews in Fisheries Science & Aquaculture. 2019. Р. 1—27. doi: 10.1080/23308249.2019.1643288
25. Hoseinifar S.H., Sun Y., Wang A., Zhou Z. Probiotics as means of diseases control in aquaculture, A Review of current knowledge and future perspectives // Frontiers in Microbiology. 2018. № 9. P. 2429. doi: 10.3389/fmicb.2018.02429
26. Van Doan H., Hoseinifar S.H., Tapingkae W., Seel-audom M., Jaturasitha S., DawoodM.A., et al. Boosted growth performance, mucosal and serum immunity, and disease resistance nile tilapia (Oreochromis niloticus) fingerlings using corncob-derived xylooligosaccharide and lactobacillus plantarum CR1T5 // Probiotics and antimicrobial proteins. 2019. P. 1—12. doi: 10.1007/s12602-019-09554-5
27. Dawood M.A.O., Koshio S., Esteban M.A. Beneficial roles of feed additives as immunostimulants in aquaculture: a review // Reviews in Aquaculture. 2018. Vol. 10. № 4. Р. 950—974. doi: 10.1111/raq.12209
28. Abdel-Tawwab M., Abdel-Rahman A.M., Ismael N.E.M. Evaluation of commercial live bakers' yeast, Saccharomyces cerevisiae as a growth and immunity promoter for Fry Nile tilapia, Oreochromis niloticus (L.) challenged in situ with Aeromonas hydrophila // Aquaculture. 2008. Vol. 280. № 1—4. Р. 185—189. doi: 10.1016/j.aquaculture.2008.03.055
29. Ringo E., Dimitroglou A., Hoseinifar S.H., Davies S.J. Prebiotics in Finfish: an update // Aquaculture nutrition: gut health, probiotics and prebiotics / Merrifield D.L, Ringo E., eds. Oxford : John Wiley & Sons, 2014. Р. 360—400. doi: 10.1002/9781118897263.ch14
30. Hoseinifar S.H., Mirvaghefi A., Merrifield D.L. The effects of dietary inactive brewer's yeast Saccharomyces cerevisiae var. ellipsoideus on the growth, physiological responses and gut microbiota of juvenile beluga (Huso huso) // Aquaculture. 2011. Vol. 318. № 1—2. P. 90— 94. doi: 10.1016/j.aquaculture.2011.04.043
31. Mahious A.S., Gatesoupe F.J., Hervi M., Metailler R., Ollevier F. Effect of dietary inulin and oligosaccharides as prebiotics for weaning turbot, Psetta maxima (Linnaeus, C. 1758) // Aquaculture International. 2006. Vol. 14. № 3. Р. 219—229. doi: 10.1007/s10499-005-9003-4
32. Akrami R., Ghelichi A., Manuchehri H. Effect of dietary inulin as prebiotic on growth performance and survival of juvenile rainbow trout (Oncorhynchus mykiss) // Journal of marine science and technology research. 2009. Vol. 4. № 3. Р. 1—9.
33. Li X., Ringo E., Hoseinifar S.H., Lauzon H.L., Birkbeck H., Yang D. The adherence and colonization of microorganisms in fish gastrointestinal tract // Reviews in Aquaculture. 2019. Vol. 11. № 3. P. 603—618. doi: 10.1111/raq.12248
34. Hoseinifar S.H., Mirvaghefi A., Amoozegar M.A., Merrifield D., Ringe E. In vitro selection of a synbiotic and in vivo evaluation on intestinal microbiota, performance and physiological response of rainbow trout (Oncorhynchus mykiss) fingerlings // Aquaculture Nutrition. 2015. Vol. 23. № 1. Р. 111—118. doi: 10.1111/anu.12373
35. Reza A., Abdolmajid H., Abbas M., Abdolmohammad A.K. Effect of dietary prebiotic inulin on growth performance, intestinal microflora, body composition and hematological parameters of juvenile beluga, Huso huso (Linnaeus, 1758) // Journal of the World Aquaculture Society. 2009. Vol. 40. № 6. Р. 771—779. doi: 10.1111/j.1749-7345.2009.00297.x "
Об авторах:
Хосейнифар Сейед Хоссейн — факультет наук о жизни и окружающей среде, Политехнический университет Марке, Италия, 60100, Анкона, Пьяцца Рома, 22; отделение животных и водных наук, факультет сельского хозяйства, Университет Чиангмая, Таиланд, 50200, Чиангмай, Су Тхеп; e-mail: hossein.hoseinifar@gmail.com; ORCID: 0000-0002-0210-9013
Доан Хиен Ван — факультет наук о животных и водных животных, сельскохозяйственный факультет, Университет Чиангмая, Таиланд, 50200, Чиангмай, Су Тхеп А шури Гасем — факультет наук о жизни и окружающей среде, Политехнический университет Марке, Италия, 60100, Анкона, Пьяцца Рома, 22
