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Effect of Commercially Available Probiotics Lactobacillus rhamnosus GG on the Body Composition of Grass Carp Fingerlings

PJZ_56_5_2189-2194

Effect of Commercially Available Probiotics Lactobacillus rhamnosus GG on the Body Composition of Grass Carp Fingerlings

Maqsood Jan1, Muhammad Zubair Anjum1*, Zahir Muhammad1, Misbah Farooq1, Muhammad Qayash Khan2 and Shamim Akhter1

1Department of Zoology, Wildlife and Fisheries, PMAS-Arid Agriculture University Rawalpindi, 46300, Pakistan

2Department of Zoology, Abdul Wali Khan University, Mardan, Khyber Pakhtunkhwa, Pakistan

ABSTRACT

The objective of current study was to evaluate the effect of commercially prepared probiotics (PREPRO) containing Lactobacillus rhamnosus GG on body composition of grass carp (fingerlings). Grass carp is a fresh water fish of edible test and having multiple feed sources. The study was aimed to enhance the production and improve the meat quality of a grass carp in controlled environment. A total of 120 grass carp (fingerlings) were selected and stocked in 12 glass aquaria (n=10 fish in each) in a triplicate manner with four different diet groups D1 (control), D2 (2g of L. rhamnosus/kg), D3 (4g of L. rhamnosus/kg), D4 (6g of L. rhamnosus/kg) as feed additive. Statistical analysis was done by using one-way analysis of variance and was applied the Duncan’s multiple range test for the identification of significant differences within the treated groups by using statistical package for the social sciences (SPSS) computer software. After the 90 days of experimental trial the results revealed that probiotics treated groups had significantly (P < 0.05) better meat quality as compared to control. The best meat quality was observed in D4 in terms of crude protein, crude fat, ash content and total moisture (45.86±0.44, 33.59±0.28, 8.80±0.34, 63.98±0.01) compared to control and other tasted groups. The L. rhamnosus improves the meat quality of grass carp.


Article Information

Received 24 January 2023

Revised 05 May 2023

Accepted 25 May 2023

Available online 26 July 2023

(early access)

Published 19 July 2024

Authors’ Contribution

SA and MZA designed the study. MJ and MF performed the research work and compile the data. MZA supervised the experiment. ZM and MQK analyzed the data. MJ processed the data and wrote manuscript.

Key words

Grass carp, Probiotics, Lactobacillus rhamnosus GG

DOI: https://dx.doi.org/10.17582/journal.pjz/20230124170136

* Corresponding author: [email protected]

0030-9923/2024/0005-2189 $ 9.00/0

Copyright 2024 by the authors. Licensee Zoological Society of Pakistan.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).



INTRODUCTION

Grass carp (Ctenopharyngodon idella) is a herbivore fresh water fish species belonging to the family Cyprinidae. It is considered to be one of the largest members of the family. It is native to East Asian lakes and large rivers from temperate to sub-tropical region in the world and its native countries are Vietnam, China and southern Russia (Shireman and Smith, 1983). In Pakistan it has been introduced from china to control over the aquatic weeds in lakes and other aquatic bodies (Khan et al., 2004).

C. idella is considered to be an important fish because of rapid growth, taste, nutritional value and aquatic weeds control (Cai and Curtis, 1989). C. idella in 2019 contributed 5.7 million tons which is 10.5% of the total freshwater aquaculture global production and was the highest fish yield among all economical fish species (FAO, 2020).

Aquatic diseases, and low fish production were the profound challenges faced by the aquaculture industry (Kuebutornye et al., 2020). The percentage infections in fishes are 54.9% bacterial, 22.6% viral, 3.1% fungal and 19.4% are from other parasitic agents (Dhar et al., 2014). To overtake this problem vaccines and antibiotics were used but commercially available vaccines were expensive and were not effective against multiple pathogenic diseases. Besides that, by using antibiotics more harmful mutated strains of bacteria were produced (Dadar et al., 2016; Kawser et al., 2019). Therefore, antibiotics and vaccines were replaced by the probiotics (Hoseinifar et al., 2017). Probiotics are living microorganisms that when administered in an adequate amount improve host health (Kuebutornye et al., 2019). Probiotics can also be defined as live microorganism that are added to food or water to be ingested in the gastrointestinal tract (GIT), where they work to balance out the body’s microbial population and promote health (Gatesoupe, 1999). Probiotics can be provided as supplement even during early stages of fish (Muroga et al., 1987). Major taxa of probiotics that have been used as a probiotics included Pediococcus, Lactobacillus, Bacillus, Enterococcus, Micrococcus, Lactococcus and yeasts (such as Aspergillus oryzae and Saccharomyces cerevisiae) (Kuebutornye et al., 2019). Probiotics improve growth rate, survival rate, gut-microbiota, and inhibit pathogen growth (Grimoud et al., 2010). Probiotics also enhance innate immunity, maintain water quality, improve meat quality, and broadly utilize in dairy food products (Duan et al., 2019; Buffie and Pamer, 2013; Perales-Puchalt et al., 2018; Yi et al., 2019; Yih et al., 2019; Xia et al., 2020; Grande et al., 2017). Probiotics also increasing villus length promoting feed utilization by increasing digestibility through production of digestive enzymes and also absorption (Thiam et al., 2015; Xia et al., 2018).

Lactobacillus are non sporolating, anaerobic, fermentative, rod shaped bacteria (El-Shall et al., 2020; Alagawany et al., 2018). They are found in variety of habitat but prefer carbohydrate rich substrate such as spoiled food, fermented milk, human mucosal membrane, plants and their derived materials (Verse and Schrezenmeir, 2008). Most of lactobacillus species produce antimicrobial substances such as (hydrogen peroxides, antimicrobial peptides, and organic acids) which constrain pathogen growth and enhanced the performance of beneficial gastrointestinal microbiota of the host (Fooks and Gibson, 2002; Vieira et al., 2008; Abdel-Latif et al., 2020).

Lactobacillus rhamnosus GG has been reported to improve body weight, meat quality, RBCs count, hemoglobin level, WBCs and the significant number of neutrophils in rabbit (Kadja et al., 2021; Simonova et al., 2008). In Pakistan varieties of probiotics have been used in aquaculture with different combinations and concentrations but there is lack of data on the effect of probiotics on grass carp. Thus the present study was undertaken to evaluate the effect of commercially available probiotics L. rhamnosus GG on meat quality of grass carp.

MATERIALS AND METHODS

Study site and fingerlings collection

The experimental trial was done at the Aquaculture and Fisheries laboratory, Department of Zoology, Wildlife and Fisheries, Pir Mehr Ali Shah Arid Agriculture University Rawalpindi. The grass carp fingerlings were obtained from the Mahseer Fish Hatchery, Garyala Attock.

Acclimatization

Before the start of experiment, fish were acclimatized in glass aquaria of (1×1×1.5 ft3) size for 7 days. The aquaria were well equipped with continuous aeration system and commercially available floating fish feed was used of (2mm) (Marine Grow Fish Feed; Hi-Tech Feeds Private Limited, Pakistan) containing 30% of crude proteins. The commercially prepared probiotic (PREPRO having ≥ 5×109 CFU of Lactobacillus rhamnosus) in powder form was purchased from the local pharmacy.

Experimental setup and diet

Fish were divided in twelve glass aquaria with 10 fingerlings per aquarium in a triplicate manner. The initial mean body weight 3.48±0.51 g. Fish were fed with four different experimental diet i.e., D1 (only supplementary fish feed), D2 (L. rhamnosus GG 2g/kg as feed additive), D3 (L. rhamnosus GG 4g/kg as feed additive) and D4 (L. rhamnosus GG 6g/kg as feed additive). Fish were fed @ 5% body weight for 90 days. On daily basis aquaria were siphoned to remove waste material of fish and the remaining feed material and was replaced with 50% of fresh water.

Proximate composition

At the end of feeding trial, fish were randomly selected from each treatment group for meat quality assessment. The viscera, fins, scales and heads were removed. Meat quality of fish were observed by proximate analysis. Crude protein, crude fat, crude ash and total moisture percentage were determined according to the standard procedures of AOAC. The moisture content was determined by drying the samples in oven at 60 ºC for one hour in dry dishes.

The moisture content were calculated as follows.

For determining crude protein Kjeldahl technique was used. For the determination of crude fat Soxhlet technique was used for 8 h in petroleum ether while total ash was done at 550 oC in a muffle furnace from the 5 g of sample for 12 h.

Statistical analysis

The obtained data was analyzed by using one-way analysis of variance (ANOVA) and the Duncan’s multiple range test (DMRT) was applied for the identification of significant differences within the treated groups by using statistical package for the social sciences (SPSS) software. The proposed results were presented as means ± standard deviation and the level of significant was set at p ≤ 0.05.

 

Table I. Effect of commercial probiotics on proximate composition (mean±standard deviation) of grass carp.

Treatments

D1

D2

D3

D4

Crude protein

40.50±0.52d

41.86±0.57c

43.33±0.70b

45.86±0.44a

Crude fats

37.08±0.71a

35.93±0.05b

35.33±0.30b

33.59±0.28c

Total ash

11.10±0.10a

10.10±0.10b

9.46±0.05c

8.80±0.34d

Moisture

75.81±0.05a

65.39±0.05c

65.99±0.00b

63.98±0.01d

 

Means ± standard deviation in the same row with different superscripts are significantly different (P < 0.05). D1, control group; D2, 2g L. rhamnosus/kg; D3, 4g L. rhamnosus/kg; D4, 6g L. rhamnosus/kg.

 

RESULTS and Discussion

Proximate composition

After 90 days of experimental trial proximate composition of crude proteins, crude fat, total ash and moisture of C. idella was analyzed (Table I). Significantly higher moisture content was found in D1 (75.81±0.05) followed by D2, D3 and the lowest was found in D4. Crude protein was found significantly increased (P<0.05) in D4 (45.86±0.44) followed by D3, D2 and D1. The significantly higher (P<0.05) percentage of crude fat was found in control group D1 (37.08±0.71) followed by D2, D3 and the lowest crud fat was found in D4. Ash content was found as highest in control group D1 (11.10±0.10) followed by D2, D3 and significantly lower (P<0.05) recorded in D4.

Probiotics are live microorganisms that can be given to fish as a food supplement to improve their health and growth performance. Probiotics provide various advantages to fish, including increased feed efficiency, improved immunity, and flourishing intestinal microbiota (Akhter et al., 2015; Huyssnh et al., 2017; Wang et al., 2018). Probiotics (L. rhamnosus GG, Bifidobacterium animalis subsp. Lactis BB-12) used as a feed additive have been found to have significantly positive effect on the proximate composition of rabbit just as we noted in this study (Kadja et al., 2021). Other studies have also reported significantly higher (17.51%) protein content and lower (4.82%) fat content after supplementing rainbow trout feed with probiotics Lactobacillus rhamnosus GG (Hooshyar et al., 2020). Vitamin A has also been used as feed additive for grass carp. The results indicated that vitamins A supplemented at 1798 IU/kg showed significant increase in crude proteins free amino acids (methionine, glutamic acids, lysine, threonine and arginine) in the muscles. Crude fat, ash and total moisture of grass carp were significantly decreased compare to that of control group (Wu et al., 2022). In our study the same fish showed significant increase in crude protein and decrease in the other parameters when its feed was supplemented with Lactobacillus rhamnosus GG.

The current result of protein and crude fat were likewise with the previously reported by Hassaan et al. (2014). The crude protein value showed significantly higher values is probably due to (1) increased intake of fish feed, (2) improved digestion and (3) higher nutritional absorption in the body (Abdel-Tawwab et al., 2008). Along with this, synthesis of protein and its deposition in muscle tissues is also increased. This could help researchers figure out the best conditions for raising Nile tilapia fingerlings (Oreochromis niloticus), having best chemical composition of fish meat in terms of higher protein contents (Jim et al., 2017).

Bisht et al. (2012) have reported that C. carpio fed at 4×108 cells 100/g B. subtilis supplemented rice bran-based diet had lower moisture content (6.75%), while protein and lipid contents (30% and 8%) were higher as compared to control group. Similarly, Suprayudi et al. (2017) recorded higher protein and lipids absorption in the body of Nile tilapia upon feeding of diet supplemented with 0.25 and 0.5 g kg-1 of dietary probiotics.

Hoyoux et al. (2009) reported that the diet containing 1×107 CFUg-1 probiotics for fish showed highest protein, fat and gross energy contents in Nile tilapia. Sahandi et al. (2019) observed improved apparent digestibility coefficient of crude protein (68.33%), crude fat (9.55%) while using 1 × 107 of two probiotics strains of Bifidobacterium. In contrast Ayoola et al. (2013) found that the control diet has the highest moisture (8.20%), fat (12.7%), and ash (4.34%) values, whereas the Clarias gariepinus group fed 1g commercially manufactured probiotics (Lactobacillus and Bifidobacterium) had the highest crude protein (66%).

CONCLUSION

In the current study it has been concluded that the use of commercial probiotics (L. rhamnosus GG) in high amount/concentration having better meat quality (high level of crude protein, low level of ash, moisture and crude fat) results compared to that of control and other groups.

Funding

No funds were provided for this project.

IRB approval

The study was approved by the institutional review board of PMAS Arid Agriculture University, Rawalpindi.

Ethical statement

We have adopted all the rules and regulations approved by the ethical committee of Pir Mehr Ali Shah Arid Agricultural University Rawalpindi in the research work.

Statement of conflict of interest

The authors have declared no conflict of interest.

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