Effect of Nano-Se Particles Supplemented Sunflower Meal Based Diets on Mineral Absorption and Carcass Composition of Cirrhinus mrigala Fingerlings

Nisar Ahmad1, Syed Makhdoom Hussain2,*, Azhar Rasul2, M. Mudassar Shahzad3, Arshad Javid4, Hamda Azmat5, M. Zubair ul Hassan Arsalan2, Sadia Tabassum2 and Bilal Ahmad2

1Department of Zoology, DG Khan Campus, University of Education, Dera Ghazi Khan, Pakistan

2Department of Zoology, Government College University, Faisalabad

3Department of Zoology, Division of Science and Technology, University of Education, Township, Lahore

4Department of Wildlife and Ecology, Faculty of Fisheries and Wildlife, University of Veterinary and Animal Sciences, Lahore

5Department of Fisheries and Aquaculture, University of Veterinary and Animal Sciences, Lahore

ABSTRACT

The research was conducted to estimate the efficacy of Se nanoparticles on mineral absorption and carcass composition of C. mrigala fingerlings fed nano-Se particles supplemented sunflower meal based diets. The experiment was consisted on seven test diets on the basis of supplementation of nano Se graded levels (0, 0.5, 1, 1.5, 2, 2.5 and 3 mg/kg). Chromic oxide was added as an inert marker. Fingerlings were fed at the rate of 5% of their live wet weight. Maximum improvement in mineral absorption (Ca, Na, K, Cu, P and Al) was observed at test diet with 1.5 mg/kg supplementation of Se nanoparticles. Maximum Fe, Mn and Cr absorption was noted at 2 mg/kg supplementation of nano Se. The highest absorption of Mg and Zn was found in the fingerlings fed at test diet with 1 mg/kg supplementation of nano Se particles. The best results in regard of body composition (CP; 61% and EE; 14%) were noted in fingerlings when fed 1.5 mg/kg levels of Se-nano particles based diets. It was concluded from the results of current study that supplementation of Se NPs (1.5 mg/kg) in sunflower meal based diet improves the mineral absorption and body composition of C. mrigala fingerlings.

Article Information

Received 21 April 2019

Revised 15 July 2019

Accepted 02 February 2021

Available online 07 May 2021

(early access)

Published 14 February 2022

Authors’ Contribution

NA conducted the trial and collected data. SMH planned and supervised the research and provided all materials for the research. NA prepared manuscript while AR and MMS helped him. AJ helped in statistical analysis. HA proffread the manuscript. MZH and ST helped in feed analysis and compiling the results. BA helped in chemical analysis of fish.

Key words

Nano-Se particles, Mineral absorption, Carcass composition, C. mrigala.

DOI: https://dx.doi.org/10.17582/journal.pjz/20190421160425

* Corresponding author: drmakhdoom90@gmail.com

0030-9923/2022/0003-1103 $ 9.00/0

Copyright 2022 Zoological Society of Pakistan

Introduction

Nanomaterials show novelty in high surface activity leading to absorption efficiency (Pelyhe and Miklos, 2013). Selenium is an important micronutrient (Dare et al., 2001). Se improves animal health and productivity and its deficiency causes great losses in livestock (Xun et al., 2012). Selenium is necessary for functioning of body and metabolism (Hamilton, 2004). Se improves animal health and productivity (Xun et al., 2012). Growth is regularized by Se and it also plays an important role in immune system activities (Pelyhe and Miklos, 2013). se improves RBCs count and the immunity of fish (Sadeghian et al., 2012; Le et al., 2013). A number of studies suggest the efficiency of Se-NPs as a dietary source (Wang et al., 2007; Zhou et al., 2009). Animals consuming a Se deficient diet face pneumonia, infertility and oxidative stress (Pelyhe and Miklos, 2013).

Fish farming is one of the best industries to explore the nano products but unfortunately a very minute work has been done in this regard. The nanomaterials hold different properties from larger molecules regarding absorption (Albrecht et al., 2006; Wang et al., 2007). Nanoparticles can be used to improve feed quality. Nanoparticles improve the absorption of feed molecules (Bunglavan et al., 2014). Nanoparticles may increase the bioavailability of nutrients. Nanoform of supplementation also increases the absorption and utilization of minerals (Vijayakumar and Balakrishnan, 2014).

C. mrigala is one of the most important fish with a maximum market demand and it contributes along with Catla catla and Labeo rohita about 67% of total freshwater fish production in South India (Krishnaveni et al., 2013). Sunflower meal contains about 45-48% crude protein (Mushtaq et al., 2006). It is used in feed formulations as it contains endogenous proteolytic enzymes to digest proteins for fish (Kocher et al., 2000). In Pakistan it has lowest cost among protein sources (Khan et al., 2006). It is important to formulate nutritionally balanced and highly palatable feed which results in maximum growth of fish (Afzal et al., 2004; Tahir et al., 2008).

In under developed countries human population is being increased day by day resulting in an alarming increase in demand of food and quality nutrition. Aquaculture is one of the best industries to fulfil such huge demand but it also requires proper feeds for fish in tanks and ponds. Though aquaculture is a very advance industry but a lot is required to produce cost effective and environment friendly feeds for aquatic organisms as a challenge. About 50-60% of the total cost of a fish form is subjected to feed (Essa et al., 2004). Fish meal is an excellent protein source for fish feed because it contains required amino acids profile and all other nutrients but it is very costly (Zhou et al., 2004). Due to high cost, uneven supply and increasing demand of fish meal it has become necessary to search for alternative protein sources (Pham et al., 2008). This is the reason we search alternative sources of protein for fish feed and sunflower is one of the best protein source from plant by-products as they are easily available round the year. These plant by-products are being used by scientists from last two decades in fish feed (Wang et al., 2015). Due to all these above said reasons we selected sunflower meal as alternative of fish meal for current study.

The major goal of the current study was to estimate the effects of Se nano-particles (optimum level) on mineral absorption and carcass composition of C. mrigala fingerlings.

Materials and methods

C. mrigala fingerlings were purchased from Government Fish Seed Hatchery Faisalabad. Fingerlings were acclimatized for 15 days in Fish Nutrition Laboratory, GCUF and were fed on the basal diet once in a day (Allan and Rowland, 1992).

Analysis of feed ingredients and Cr-nanoparticles

The feed ingredients were analysed by following standard methods (AOAC, 1995). Se nenoparticles (NPs) were purchased from market (sigma-Aldrich), to confirm their pure crystalline structure and size, they were analysed by XRD and TEM (TEM-JEOL2100-20171206), respectively (Ramamurthy et al., 2013; Iqbal et al., 2014).

Formation of pellets

All the feed ingredients were grinded until they passed through 0.5 mm sized sieve. All ingredients were mixed for 5 min, fish oil was gradually added. Finally water was added slowly to make suitable dough and pellets were formulated thereafter with the help of pelleting machine by following Lovell (1989).

Preparation of NP stock solution

Preparation and confirmation of stock solutions of NPs was carried out according to Federici et al. (2007). Stock solution of 100% pure NPs dry powder was made by sonication method (for 6-8 h) and from this stock solution further dilutions were made to ensure our required levels (0, 0.5, 1, 1.5, 2, 2.5 and 3 mg/kg) of Se NPs.

Addition of NPs to basal diets

The diluted Se solutions were sonicated further for 15 min just before spraying on basal diets according to Ramsden et al. (2009). One Kg feed was placed in a commercial food mixer and gradually sprayed with the appropriate dilution. The seven test diets were formulated by mixing graded levels (0, 0.5, 1, 1.5, 2, 2.5 and 3 mg/kg) of nano Se. NPs immediately coated the feed pellets. The feed pellets were allowed to dry then ultimately were stored in air tight containers for further use.

Feeding protocol and sample collection

C. mrigala fingerlings were fed for 90 days on prescribed diets mentioned above at the rate of 5% of their live wet weight. Three replicates were used for each diet and 15 fingerlings were stocked in each replicate. Their feces were collected by alternative opening and closing of valve-I and valve-II following Hussain et al. (2018).

Chemical analysis of feed and feces

The samples of experimental diets and feces were homogenized using mortar and pestle and analysed following standard methods (AOAC, 1995). Diets and feces samples were digested in boiling nitric acid and perchloric acid mixture (2:1) by following standard methods (AOAC, 1995). After appropriate dilution, mineral contents such as calcium (Ca), magnesium (Mg), iron (Fe), copper (Cu), zinc (Zn) and manganese (Mn) were estimated using Atomic Absorption Spectrophotometer (Hitachi Polarized Atomic Absorption Spectrometer, Z-8200). Calibrated standards for mineral estimation were prepared from commercially available standards (AppliChem® Gmbh Ottoweg4, DE-64291 Darmstadt, Germany). The estimation of Na and K was done through flame photometer (Jenway PFP-7, UK). Phosphorus (P) was analysed colorimetrically (UV/VIS spectrophotometer) using ammonium molybdate as reagent at 720 nm absorbance through standard methods (AOAC, 1995). Chromic oxide contents in diets and feces were estimated after oxidation with molybdate reagent using a UV-VIS 2001 Spectrophotometer at 370 nm absorbance (Divakaran et al., 2002).

Calculation of apparent digestibility coefficient (ADC)

Apparent mineral digestibility coefficients (ADC) of test diets were calculated by the standard formula (NRC, 1993).

 

Table I.- Chemical composition (%) of feed ingredients.

Ingredients	Fish meal	Rice polish	Wheat flour	Sunflower
Dry matter (%)	91.67	94.06	92.4	93.80
Crude protein (%)	49.03	11.87	09.73	40.81
Crude fat (%)	6.93	12.69	2.24	3.69
Crude fiber (%)	1.23	11.91	2.73	1.94
Ash (%)	23.15	11.32	1.99	09.96
Gross energy (kcal/g)	2.49	3.41	3.06	3.64
Carbohydrates	19.66	52.21	82.21	43.6

 

Chemical analysis of fish whole body

At the end of 90 days trial four fish from each tank were sacrificed randomly and dried at room temperature. Moisture contents of carcass were calculated after oven-drying of homogenised samples at 105oC for 12 h. Micro kjeldahl apparatus (InKjel Mbehr Labor Technik GmbH D-40599 Dusseldorf) was used to determine the crude protein (CP) (N × 6.25) whereas soxhlet system (Soxhlet Extraction Heating Mantels, 250 ml 53868601) was used to check the amount of crude fat by the help of petroleum ether extraction (EE) method. Crude fiber contents were calculated as loss on ignition of dried lipid-free residues after digestion with 1.25% H2SO4 and 1.25% NaOH whereas ash was determined by ignition at 650oC for 12 h in electric furnace (Naberthern B170) to constant weight. Total carbohydrates (N-free extract) were determined by difference, i.e., total carbohydrates % =100- (EE % + CP % + Ash % + CF %). Oxygen bomb calorimeter was used to estimate the gross energy.

Statistical analysis

One-way analysis of variance was applied to data of mineral absorption and carcass composition of fish (Steel et al., 1996). Tukey’s Honesty Significant Difference Test (p<0.05) was used to compare the differences among various levels (Snedecor and Cochran, 1991). For statistical analysis CoStat Computer Package (Version 6.303, PMB 320, Monterey, CA, 93940 USA) was used.

 

Results

Results of morphological analysis of the Se-NPs by using transmission electron microscope (TEM) are shown in Figure 1. TEM confirms the shape and size of Se NPs, as magnified form depicted in Figure 1A and normal TEM form in Figure 1B. In these samples, the scale bar was set to 10 nm in case of magnified TEM image and 50 nm of scale bar in terms of normal form TEM image. TEM image justify the spherics form of Se NPs with about 8–10 nm diameter almost homogenous structure format. The above results of TEM confirm that Se NPs used in experimental diets of our study contain size less than 100nm about 10nm. It confirms that they are pure nano particles in their nature.

The crystal structure and the phase composition of selenium nanoparticles were confirmed by using X-Ray Diffraction (XRD) techniques shown in Figure 2. The XRD pattern confirms very clearly that the sample is nano-crystalline in nature as it matches very well with that of the standard selenium powder of selenium nano-particles.

Analysis showed that there was balanced amount of all the minerals in the control and test diets formulated by sunflower meal based diet supplemented with Se NPs at 0, 0.5, 1, 1.5, 2, 2.5 and 3 mg/kg level. Amount of minerals such as Ca, Na, K, Fe, Cu, Zn, Mn, P, Mg, Al and Cr were statistically (p<0.05) similar in control and Se nano supplemented test diets whereas Ba, Cd, Co and Ni were found lowest from the range (<0.0001) in diets.

Mineral absorption was lowest at control diet (0 mg/kg Se NPs level based diet) after that it started to increase from 0.5 mg/kg level and reached its maximum at 1.5 or 2 mg/kg level. It was also noted that further increase in Se NPs supplementation decreased absorption of minerals (Table III). Maximum mineral absorption i.e. Ca (70%), Na (74%), K (70%), Cu (66%), P (75%) and Al (61%) were found at 1.5 mg/kg level diet. Whereas highest absorption value of Fe (67%), Mn (71%) and Cr (57%) was noted at 2 mg/kg level diet. On the other side Mg (64%) and Zn (62%) were absorbed at 1 mg/kg level diet. These values were statistically higher (p<0.05) when compared with control and other Se-nano supplemented test diets. It was also observed that some of the minor minerals such as Ba, Cd, Co and Ni were very low (<0.0001) in diet and could not be analysed when absorption was calculated.

It was concluded that supplementation of Se-nano particles in sunflower meal based diets played a significant role in improving mineral absorption. These reduced minerals in water will be helpful to control water pollution. Furthermore, Se NPs supplementation in fish feed may also decrease feed cost because there will be no need of extra mineral supplementation.

Similarly maximum values of CP (61%) and EE (14%) were observed in fish fed at 1.5 mg/kg Se-nano diet. The least CP (54%) contents in fish body was observed when fed test diet T7, 3 mg/kg Se-nano and the least EE (9%) contents in fish body was observed when fed test diet T2, 0.5 mg/kg Se-nano. From results (Table IV) it was found that improvement in protein and fat contents for C. mrigala fingerlings was started from 0.5 mg/kg Se-nano diet and reached its maximum in fingerlings fed 1.5 mg/kg level of Se-nano supplemented sunflower meal based diet. Further increase in Se-nano supplementation could not enhance nutrient contents in fish body.

Analysis showed minimum amount of carbohydrates (11%) and crude fiber (1%) contents in fish fed at 1 and 1.5 mg/kg Se-nano level based diets. However maximum values of carbohydrates (18%) and crude fibre (1%) were observed in fish fed at 0 mg/kg Se-nano level based diet (control diet). Minimum ash contents (8%) and moisture contents (5%) were recorded in fish fed at 1.5 mg/kg Se-nano level based diet that were significantly (p<0.05) different from the fish which fed at other Se-nano supplemented diets and without supplemented sunflower meal based diet (control diet). However maximum moisture (7%) and ash (10%) contents were found in the fish fed at control diet and 3 mg/kg Se-nano level based diet, respectively. It was found that 1.5 mg/kg Se-nano supplementation in diet is the best level for the maximum deposition of protein and lipids in fish; these nutrients are essential for fish performance. Moreover, almost lowest carbohydrates, ash, crude fiber as well as moisture contents were observed in C. mrigala fed 1.5 mg/kg Se-nano diet.

 

Table II.- Ingredients composition (%) oilseed meal based test diets.

Ingredients	Test diet-I	Test diet-II	Test diet-III	Test diet-IV	Test diet-V	Test diet-VI	Test diet-VII
Nanoparticles (mg/kg)	0	0.5	1	1.5	2	2.5	3.0
Sunflower meal	50	50	50	50	50	50	50
Fish meal	14.5	14.5	14.5	14.5	14.5	14.5	14.5
Wheat flour*	13	13	13	13	13	13	13
Rice polish	11	11	11	11	11	11	11
Fish oil	7.5	7.5	7.5	7.5	7.5	7.5	7.5
Vitamin premix**	1	1	1	1	1	1	1
Minerals premix***	1	1	1	1	1	1	1
Ascorbic acid	1	1	1	1	1	1	1
Chromic oxide	1	1	1	1	1	1	1

 

*Nano-particles will be added on the cost of wheat flour. ** Vitamin premix/kg: Vitamin D3, 3,000,000 IU; Vitamin A, 15,000,000 IU; Vitamin E, 30000 IU; Vitamin B1, 3000 mg; Vitamin B6, 4000 mg; Vitamin B12, 40mg; Vitamin B2, 7000 mg; Vitamin C, 15,000 mg; Vitamin K3, 8000 mg; Folic acid, 1500 mg; Calcium pantothenate, 12,000mg; Nicotinic acid, 60,000 mg. ***Mineral premix/kg: Mn (manganese), 2000 mg; Ca (calcium), 155 gm; Zn (zinc), 3000 mg; Cu (copper), 600 mg; Co (cobalt), 40 mg; I (iodine), 40 mg; P (phosphorous), 135 gm; Fe (iron), 1000 mg; Mg (magnesium), 55 gm; Se (selenium), 3 mg; Na (sodium), 45 g.

 

Table III.- Apparent mineral digestibility (%) of C. mrigala fingerlings fed graded levels of Se-nano supplemented sunflower meal based diets.

Diets	Se-nano (mg/kg)	Concentrations (%)
		Diet	Feces	Digestibility
Digestibility of Ca
Test diet – I (control diet)	0	0.87	0.48 a	49.27 e
Test diet – II	0.5	0.88	0.47 a	50.47 e
Test diet – III	1	0.89	0.38 bc	59.95 c
Test diet – IV	1.5	0.87	0.29 c	69.52 a
Test diet – V	2	0.88	0.31 c	68.00 b
Test diet – VI	2.5	0.88	0.44 ab	55.05 d
Test diet – VII	3	0.88	0.50 a	49.61 e
PSE		0.0385	0.018	0.309
P-value		0.99 NS	0.00***	0.00***
Digestibility of Na
Test diet – I (control diet)	0	0.008	0.004a	50.26 c
Test diet – II	0.5	0.008	0.004ab	52.05 c
Test diet – III	1	0.008	0.003bc	59.40 b
Test diet – IV	1.5	0.008	0.002d	73.95 a
Test diet – V	2	0.008	0.002d	72.11 a
Test diet – VI	2.5	0.008	0.003c	60.59 b
Test diet – VII	3	0.008	0.003bc	60.74 b
PSE		0.0002	0.0001	0.406
P-value		0.99 NS	0.00***	0.00***
Digestibility of K
Test diet – I (control diet)	0	1.403	0.822a	46.52e
Test diet – II	0.5	1.390	0.753ab	50.14d
Test diet – III	1	1.400	0.500c	66.80b
Test diet – IV	1.5	1.386	0.457c	70.24a
Test diet – V	2	1.396	0.505c	67.57b
Test diet – VI	2.5	1.393	0.717b	53.13c
Test diet – VII	3	1.393	0.759ab	51.45d
PSE		0.026	0.015	0.329
P-value		0.99 NS	0.00***	0.00***
Digestibility of P
Test diet – I (control diet)	0	2.023	1.044a	52.89d
Test diet – II	0.5	2.026	0.870b	60.51c
Test diet – III	1	2.014	0.660c	69.52b
Test diet – IV	1.5	2.017	0.565d	74.67a
Test diet – V	2	2.010	0.650c	70.97b
Test diet – VI	2.5	2.020	0.867b	60.92c
Test diet – VII	3	2.020	0.921b	59.33c
PSE		0.026	0.011	0.492
P-value		0.99 NS	0.00***	0.00***

Diets	Se-nano (mg/kg)	Concentrations (%)
		Diet	Feces	Digestibility
Digestibility of Fe
Test diet – I (control diet)	0	0.046	0.027a	46.03f
Test diet – II	0.5	0.047	0.026a	48.74e
Test diet – III	1	0.046	0.021bcd	58.24c
Test diet – IV	1.5	0.048	0.018cd	64.66b
Test diet – V	2	0.047	0.017d	67.13a
Test diet – VI	2.5	0.047	0.023abc	56.13cd
Test diet – VII	3	0.047	0.023ab	55.44d
PSE		0.002	0.0009	0.439
P-value		0.99 NS	0.00***	0.00***
Digestibility of Cu
Test diet – I (control diet)	0	0.005	0.0031 a	47.40 c
Test diet – II	0.5	0.005	0.0023 b	62.79 b
Test diet – III	1	0.005	0.0021 b	64.92 a
Test diet – IV	1.5	0.005	0.0021 b	66.02 a
Test diet – V	2	0.005	0.0023 b	62.10 b
Test diet – VI	2.5	0.005	0.0035 a	42.71 d
Test diet – VII	3	0.005	0.0037 a	41.45 d
PSE		0.0002	0.0001	0.375
P-value		0.99 NS	0.00***	0.00***
Digestibility of Zn
Test diet – I (control diet)	0	0.042	0.023ab	49.10d
Test diet – II	0.5	0.041	0.021abc	51.17c
Test diet – III	1	0.041	0.017c	61.87a
Test diet – IV	1.5	0.042	0.018bc	60.72a
Test diet – V	2	0.042	0.020abc	56.98b
Test diet – VI	2.5	0.043	0.024a	47.77de
Test diet – VII	3	0.043	0.025a	46.45e
PSE		0.002	0.001	0.364
P-value		0.99 NS	0.0008***	0.00***
Digestibility of Mn
Test diet – I (control diet)	0	0.024	0.014a	45.09e
Test diet – II	0.5	0.023	0.013ab	47.41d
Test diet – III	1	0.024	0.009abc	62.83c
Test diet – IV	1.5	0.025	0.008bc	69.82a
Test diet – V	2	0.024	0.007c	70.94a
Test diet – VI	2.5	0.025	0.009abc	64.99b
Test diet – VII	3	0.025	0.010abc	62.29c
PSE		0.002	0.001	0.255
P-value		0.99 NS	0.006**	0.00***
Digestibility of Mg
Test diet – I (control diet)	0	0.009	0.004bc	56.30c
Test diet – II	0.5	0.009	0.004bcd	57.16 c
Test diet – III	1	0.009	0.003d	64.37 a
Test diet – IV	1.5	0.009	0.003cd	62.16 b
Test diet – V	2	0.009	0.004bc	57.38 c
Test diet – VI	2.5	0.009	0.005ab	50.97 d
Test diet – VII	3	0.009	0.005a	48.31 e
PSE		0.0002	0.0001	0.374
P-value		0.99 NS	0.00***	0.00***

Diets	Se-nano (mg/kg)	Concentrations (%)
		Diet	Feces	Digestibility
Digestibility of Cr
Test diet – I (control diet)	0	0.028	0.016a	46.38d
Test diet – II	0.5	0.027	0.017a	42.36e
Test diet – III	1	0.027	0.015a	45.57d
Test diet – IV	1.5	0.028	0.015a	51.55c
Test diet – V	2	0.027	0.013a	57.23a
Test diet – VI	2.5	0.026	0.013a	55.41b
Test diet – VII	3	0.026	0.013a	54.69b
PSE		0.002	0.001	0.327
P-value		0.99 NS	0.153 NS	0.00***
Digestibility of Al
Test diet – I (control diet)	0	0.0006	0.0004a	36.30 e
Test diet – II	0.5	0.0006	0.0003bcd	51.66 c
Test diet – III	1	0.0006	0.0002d	60.43 a
Test diet – IV	1.5	0.0006	0.0002cd	61.13 a
Test diet – V	2	0.0006	0.0003bcd	56.09 b
Test diet – VI	2.5	0.0006	0.0003bc	47.43 d
Test diet – VII	3	0.0006	0.0003ab	46.54 d
PSE		2.211	1.756	0.242
P-value		0.98 NS	0.0001***	0.00***
Digestibility of Cd, Co, Ni and Ba
Test diet – I (control diet)	0	<0.0001	<0.0001	<0.0001
Test diet – II	0.5	<0.0001	<0.0001	<0.0001
Test diet – III	1	<0.0001	<0.0001	<0.0001
Test diet – IV	1.5	<0.0001	<0.0001	<0.0001
Test diet – V	2	<0.0001	<0.0001	<0.0001
Test diet – VI	2.5	<0.0001	<0.0001	<0.0001
Test diet – VII	3	<0.0001	<0.0001	<0.0001
PSE		<0.0001	<0.0001	<0.0001
P-value		<0.0001	<0.0001	<0.0001

 

PSE, pooled SE = √MSE/n (where MSE is mean-squared error).

 

Table IV.- Proximate composition (%) of C. mrigala carcass fed graded levels of Se-nano supplemented Sunflower meal based diets.

Diets	Se-nano (mg/kg)	Protein	Fat	Ash	Moisture	Crude fibre	Carbohydrates
Test diet – I (control diet)	0	54.93 de	9.27 d	9.37 a	7.16 a	1.26 a	18.02 a
Test diet – II	0.5	55.41 d	9.23 d	9.34 a	7.10 a	1.26 a	17.66 a
Test diet – III	1	60.74 a	13.01 ab	8.26 bc	6.14 b	1.06 a	10.78 c
Test diet – IV	1.5	61.37 a	13.72 a	7.76 c	5.17 c	1.02 a	10.97 c
Test diet – V	2	58.28 b	12.82 b	8.42 b	5.50 c	1.21 a	13.77 b
Test diet – VI	2.5	57.29 c	11.73 c	9.44 a	6.40 b	1.24 a	13.90 b
Test diet – VII	3	54.29 e	11.73 c	9.60 a	6.40 b	1.24 a	16.73 a
PSE		0.193	0.150	0.125	0.118	0.058	0.287
P Value		0.00***	0.00***	0.00***	0.00***	0.046*	0.00***

 

Means within columns having different superscripts are significantly different at p< 0.05. Data are means of three replicates. PSE, pooled SE = √MSE/n (where MSE is mean-squared error).

 

Discussion

Our study demonstrates that Se NPs increase intestinal uptake of minerals. These findings are coinciding with the results of Srinivasan et al. (2016) who found that optimum concentration of nano-particles increased the absorption of Cu, Zn, Ca, Mg, Na and K minerals in test prawns (Macrobrachium rosenbergii post-larvae). The above said results were on supplementation of Fe2O3 nano particles at the rate of 20 mg/kg while the others levels were 10, 30, 40 and 50 mg/kg. Similarly an increase in iron has been reported in L. rohita fed with ferrous oxide NPs supplemented diets (Behera et al., 2014) and in M. rosenbergii fed with Zn and Cu nano sized forms (Muralisankar et al., 2014, 2016).

Differing from our results Zaboli et al. (2013) found no effects on composition of blood minerals (Ca, P, Fe, Cu and Zn) of Markhoz goat kids when fed to zinc oxide (ZnO) and nano zinc oxide (nZnO) supplemented diets. Zinc was administered at daily doses of zero, 20 and 40 ppm in both ZnO and nZnO groups by adding to their basal diets. Our results are antagonistic to the findings of Sirirat et al. (2012) who found that supplementation of Nano Cr Pic did not affect the absorption of Cu, Zn, Fe and Mn. Our findings are also opposed by the results of different studies who concluded that dietary Zn supplementation showed no affects on the Zn levels of blood (Droke et al., 1998; Eryavuz et al., 2002; Salama et al., 2003; Spears et al., 2004). The result may be differing due to difference in basal diet, experimental species, nature, level and shape of nanoparticles.

Our results are also parallel to the findings of Bunglavan et al. (2014), that nanoparticles improved the minerals bioavailability. Similarly, lien et al. (2009) concluded that nano Cr pic group significantly enhanced the Cr digestibility in rats.

The results of current study proved that nano particles supplementation is very important to improve the carcass composition of C. mrigala fingerlings. These results are similar to the findings of Srinivasan et al. (2016) who found that supplementation of Fe2O3 NPs significantly improved carcass parameters as compared to control diets in giant fresh water prawn. Wang and Xu (2004) reported that supplementation of Cr-Nanoparticles has beneficial effects on carcass composition, pork quality and individual skeletal muscle weight. Dietary Cr-Nano supplementation increased (p<0.05) the lean ratio of carcass of pigs by 14.06%. This improvement is due to the special metabolism pathway and deposition mechanism of NPs in carps due to which soluble proteins can interact with nanoparticles to form halo (crona). Nano-protein cronas can interfere with protein folding and can enhance protein cross linking (Zhou et al., 2009; Onuegbu et al., 2018). When concentration of NPs crosses the optimum levels then feed starts to lose palatability which may be the possible reason of decrease of carcass parameters on higher levels of supplementation (Onuegbu et al., 2018).

On the other hand, Ashouria et al. (2015) reported that proximate composition of fish was not affected by the dietary treatment after 8-weeks of culture, indicating that the carp muscle proximate composition is not sensitive to dietary Se treatments. Similar observation was also reported by Le et al. (2013) for juvenile yellowtail king fish. Wang et al. (2015) studied the effects of Cu NPs and soluble Cu on carcass composition of juvenile Epinephelus coioides. The fish were exposed in triplicate to control, 20 or 100μg CuL-1 as either copper sulphate (CuSO4) or Cu-NPs for 25 days. With an increase in CuSO4 and Cu-NPs dose, crude protein and crude lipid decreased while ash and moisture increased ultimately causing decrease in growth performance of fish.

Conclusion

This study provides sufficient evidence that supplementation of the Se nano particles are helpful for the improvement of mineral absorption and carcass composition C. mrigala fingerlings fed sunflower meal based diets. It was also concluded that 1.5 mg/kg supplementation levels of Se-nanoparticles is the optimum level for the improvement of all above said factors and higher supplementations could not cause further improvement.

Acknowledgements

The authors would like to acknowledge HEC Pakistan for providing funds for NRPU Projects # 20-4892/NRPU/R&D/HEC/14/1145 and 5649/Punjab/NRPU/R&D/HEC/2016 to conduct this research work.

Statement of conflict of interest

The authors have declared no conflict of interests.

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