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ISSN 1303-2712
Turkish Journal of Fisheries and Aquatic Sciences 13: 495-501 (2013) DOI: 10.4194/1303-2712-v13_3_12

Effects of Dietary Zeolite and Perlite Supplementations on Growth and

Nutrient Utilization Performance, and Some Serum Variables in Common
Carp, (Cyprinus carpio)
A. Khodanazary1,*, F. Boldaji2, A.Tatar2, B. Dastar2
1
Department of Fisheries, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran.
2
Department of Animal Sciences, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran.

* Corresponding Author: Tel.:+98.937 660284 ; Fax: +98.171 2245886; Received 23 July 2012
E-mail: khodanazary@yahoo.com Accepted 24 July 2013

Abstract

Zeolite and perlite are natural volcanic rocks with ion exchanging properties. This experiment was carried out to
determine the effects of different dietary levels of natural zeolite and perlite on growth performance, apparent digestibility
coefficients of dry matter and protein, ash contents of bone and scales as well as blood parameters in common carp Cyprinus
carpio. Five experimental diets tested were a control diet, and zeolite and perlite additions at levels 2.5 and 5%. Each
treatment was tested in 3 replicates tanks using fish with an initial average weight of 30±0.5 g. Fish were hand-fed, twice a
day for 8 weeks. The results showed that the dietary perlite and zeolite had no an influence on the fish condition factor
(P>0.05). There were improving effects of the dietary additions, particularly at 5 %, on growth performance of fish compared
to the control. The dry matter and protein apparent digestibility coefficients (ADC; means± S.D) for the experimental diets
were, respectively, Z 2.5 (Zeolite 2.5): 83.0±0.89 and 91.99±0.28; Z 5 (Zeolite 5): 87.48±0.76 and 95.14±0.92; P 2.5 (Perlite
2.5): 84.87±0.71 and 93.42±0.64; P 5 (Perlite 5): 87.92±1.27 and 95.80±0.57. Significantly higher ash contents were found in
bones and scales of fish fed with the experimental diets in comparison with the control diet (P<0.05). The increase of dietary
zeolite resulted in a significantly higher concentration of Ca in serum. While the zeolite and perlite diets did not affect serum
Mg, P and total protein contents of serum, a significantly enhanced glucose value was observed in fish fed Z 5. The Na and
cholesterol content of serum decreased with adding perlite and zeolite, although K content increased (P<0.05). In conclusion,
the results of this study suggest that dietary supplement of zeolite and perlite can be used as a new aquafeed ingredient for
commom carp.

Keywords: Cyprinus carpio, dietary zeolite, dietary perlite, digestibility, growth performance, blood parameters.

Introduction production (Pond and Mumpton, 1984).

Studies in Japan using zeolite as a dietary
Zeolites are crystalline, hydrated supplement for animals showed that the test animals
aluminosilicates of alkali (e.g. Na+, K+) and alkaline generally grew faster than control groups with a
(e.g. Mg+2, Ca+2) earth cations, consisting of three simultaneous decrease in the amount and cost of the
dimensional frameworks of SiO-4 and AlO-5 tetrahedra feed (Mumpton and Fishman, 1977). Dietary
linked through the shared oxygen atoms inclusion of zeolites in fish has also been reported to
(Papaioannou, et al., 2005). On the other hand, perlite improve growth and feed utilization (Mumpton and
is defined as a naturally occurring glassy volcanic Fishman, 1977; Papaioannou et al., 2005). However,
siliceous rock with a marked concentric onion-like it is difficult to make a general conclusion about the
structure of fractures, caused by concentration during performance improving effects of dietary zeolites in
cooling (Mathialagan and Viraraghavan, 2002). There aquaculture. Research data reported in the published
are various hypotheses concerning the function of literature provide evidence of a growth promoting
zeolite in diet of fish: as dietary supplements in effect when zeolites are used as additives in animal
animal diets, data in the published literature provide nutrition. The effect of dietary zeolites on feed intake
evidence of a growth promoting effect when zeolites varies in different researches. For instance, Leonard
are used as additives in animal nutrition, provide (1979) added 2% natural zeolite (clinoptilolite, the
oxygen-enriched air to aquacultural systems, have most abundant zeolite in the nature) to trout diets, and
beneficial effects in fish hatchery systems, and found a significant improvement in weight gain over a
allowing reductions in energy use and increasing 64-days feeding period. The use of clinoptilolite at 5
© Published by Central Fisheries Research Institute (CFRI) Trabzon, Turkey
in cooperation with Japan International Cooperation Agency (JICA), Japan
496 A. Khodanazary et al. / Turk. J. Fish. Aquat. Sci. 13: 495-501 (2013)
and 10% levels did not affect the growth of coho (7.5%) (Table 1). The control diet was either
salmon (Edsall and Smith, 1989). Furthermore, supplemented with 2.5 and 5 % of zeolite (named Z
Reinitz (1984) demonstrated that dietary inclusion of 2.5 and Z 5, respectively) or perlite (P 2.5 and P 5
zeolite (sodium bentonite), at 5, 10 and 15% respectively). Zeolite (clinoptilolite) used as raw
adversely affected weight gain in rainbow trout. material and expanded perlite added to experimental
Inclusion levels of zeolite into animal diets range diet. Expanded perlite is an economical insulation for
from 1 to 10 %, but the recommended level is 1 %for high and low temperature and cryogenic applications.
synthetic zeolite, and 10 % for nature zeolite Ingredients and composition of five diets are shown in
(Shariatmadari, 2008). Table 1. All ingredients were finely grinded, soybean
Therefore, the use of perlite and zeolite in fish oil, sunflower oil, molasses and deionized water
diets has not been studied extensively and the existing (30%) was added and mixed in a homemade mixer
results are inconclusive. The present experiment was (IKA® T25 digital ULTRA-TURRAX®) and then
conducted to evaluate the effects of zeolite and perlite pelleted (3.5 mm diameter) using a meat chopper.
inclusions in common carp diets on growth and Pellets were then air-dried and screened to remove
nutrient utilization performance, apparent digestibility fines before feeding. To all diets, 1% chromic oxide
coefficients of dry matter, protein as well as ash (Cr2O3) was added as an indigestible marker in order
contents of bone, scales and blood serum biochemical to measure dry matter and protein digestibility
properties. coefficients.

Materials and Methods Experimental Fish and Feeding Trial

Experimental Diets A total of 150 juvenile carp purchased from a

private fish farm were transported to the fishery
A control diet was formulated to contain 29.35% research center, Gorgan University of Agricultural
crude protein and 7.68% crude fat using wheat (10%), Sciences and Natural Resources, Iran. The carp were
corn (20%), soybean meal (60%), fish meal (26.5%), acclimatized for one month in 8 tanks (300 L
plant oil (6%) and other supplemental ingredients capacity) and then randomly distributed into 15

Table 1. Ingredients and nutrient composition of the experimental diets

Ingredients of control diet (g/kg)

Fish meal (clupea meal) 265.0
Soybean meal 300.0
Wheat meal 100.0
Corn meal 200.0
Soy oil 30.0
Sunflower oil 30.0
Vitamin premix1 7.5
Mineral premix2 7.5
Lysine 10.0
Methionine 14.0
Lysetin 2.0
CaCo3 4.0
Molasses 10.0
Salt 10.0
Cr2O3 10.0
Zeolite 0.00
Perlite 0.00
Experimental diets Control Z 2.5 Z5 P 2.5 P5
Control diet 100 97.5 95 97.5 95
Zeolite - 2.5 5 - -
Perlite - - - 2.5 5
Calculated analysis (% dry matter)
Ash 8.75 10.96 13.41 11.05 13.75
Crude protein (N× 6.25) 29.35 28.61 27.88 28.61 27.88
fat 7.68 7.48 7.29 7.48 7.29
1
Each kg of vitamin premix containing: vitamin A, 120000 IU; vitamin D3, 80000 IU; vitamin E, 8000 mg; vitamin K3, 200 mg; vitamin B1,
600 mg; vitamin B2, 1000 mg; vitamin B6, 600 mg; vitamin B12, 1600 mg, vitamin C, 10400 mg; vitamin Nicotinic acid, 6000 mg; vitamin
Calciumed pantothenate, 1800 mg; vitamin Folic acid, 320 mg; vitamin d- Biotin, 32 mg; vitamin Inositol, 4800 mg; vitamin Antioxidant,
1000 mg.
2
Each kg of mineral premix containing: Manganese, 520 mg; Copper, 120 mg; Zinc, 1200 mg; iron, 800 mg; Selenium, 10 mg; I, 40 mg; Co,
10 mg; Colin chloride, 24000 mg.
 A. Khodanazary et al. / Turk. J. Fish. Aquat. Sci. 13: 495-501 (2013) 497
experimental tanks (100 L capacity). The mean initial 10000 rpm for 10 min, and the serum was separated
weight of fish was 30±0.5 g. Health status of the fish and stored at -20 ºC before analysis. The serum was
during the acclimation and experimental period was analyzed for P, Ca, Fe, Mg, K, and Na using
good, there was no mortality during the experiment. spectrophotometric (). cholesterol and glucose
Each of the five experiment groups were tried in three concentration (Roch Germany, 1489348). Scales and
replicate tanks. The water was aerated using air-stone the central bones were also collected from the fish in
diffusers and conducted recirculating system. each tank after blood sampling. The scales were
Dissolved oxygen, temperature and pH were 6-7 mgL- sampled from the ventral muscle section. The bones
1
, 25± 0.3 ºC and 7.5, respectively. were separated from soaked fish warm (40 ºC)
During the 8 weeks experimental period, fish deionized water for 5 min and then the bone and
were hand-fed twice daily for 5 days a week (9:30 and scales were dried in an oven (105 ºC) and ashed at
14:30 h) at a rate of 3% body weight in the calculation 480 ºCfor 48 h.(Nwanna et al., 2007). Chromic oxide
of daily feeding amounts of each treatment the diet the diets and faeces samples were determined
dilution rates, because these are inert materials (Dias according to Williams et al. (1962) and chemical
et al., 1998). The fish were weighted collectively with composition analysis of diets and faeces were made
bi-weekly intervals and feeding rates were corrected for dry matter, ash, crude protein and fat, using
accordingly. No feeding was done on the sampling AOAC (1990) procedures.
days. Before weighting, fish in each tank were
anaesthetized cite clove oil (100 pm) (Anderson et al., Statistical Analysis
1997).
Data are presented as mean± standard deviation.
Fecal Collection To test differences between dietary treatments, all
data were subjected to one-way analysis of variance
At the end of experiment, all fish in each tank (ANOVA) using SPSS software (version 11).
were dissected and faeces were collected from Duncan's multiple range test was used to separate the
intestine. During the dissection the intestine was means among treatments with a probability level of
clamped immediately proximal to the anus and at the P<0.05 (Duncan, 1955).
level of pelvic fins using surgical clamps. This section
of the intestine was removed and the contents were Results and Discussion
gently squeezed into a plastic bottle. Fecal samples
were lyophilized (ALPHA 1-2 LD plus) and stored at Growth Performance
-20 °C until further analysis (Percival et al., 2001).
Growth performance of fish fed with different
Chemical Analysis experimental diets is presented in Table 2. Dietary
inclusions of zeolite and perlite had a significant
Before the termination of the experiment, blood positive effect on the growth performance of fish
samples collected after 24 h fasting. Four fish were (P<0.05), being more marked in fish fed Z 5. When
randomly captured from each tank and anesthetized the inclusion levels were considered, it was seen that
with clove oil (100 ppm). The blood samples were 5 % better than 2.5 %. The influence of the dietary
collected from caudal vein using the syringe, and then incorporations was not clear in terms of FCR and
transferred to syringed into 1.5 ml tubes (Nwanna et FCE, but the best value was observed in fish diet Z 5
al., 2007). Tubes were centrifuged (Eppendorf AG without differing from the control.
22331 Hamburg, Centrifuge 4515D, Germany) at These results are partly in agreement with the

Table 2. The effects of perlite and zeolite on growth performance of common carp fish grown over 56 days

Control Z 2.5 Z5 P 2.5 P5

Initial weight (g) 30.25±2.76 29.62±2.56 30.35±3.02 28.95±2.49 30.14±2.48
Final weight (g) 58.01±4.16c 61.62±8.16abc 66.41±10.89a 59.48±10.08bc 64.51±9.54ab
WG 27.52±2.10b 32.00±4.60ab 36.06±4.29a 34.53±3.83ab 34.37±4.24ab
WG (%) 90.42±9.74b 108.51±18.87ab 118.74±12.59a 105.71±15.70ab 113.83±6.70ab
FCR1 1.84±0.13b 2.03±0.33ab 1.77±0.19b 2.52±0.31a 2.24±0.27ab
FCE2 54.16±3.81ab 50.00±8.00abc 56.66±6.42a 40.00±5.00c 44.99±5.77bc
SGR (%/day)3 1.14±0.09b 1.30±0.15ab 1.40±0.09a 1.28±0.13ab 1.35±0.05ab
CF4 1.60±0.05a 1.64±0.09a 1.66±0.03a 1.69±0.09a 1.66±0.04a
The initial mean body weight: 30 ±0.5 g. Values are means ±S.D. of triplicate and values within the same row with different superscript
letters are significantly different (P<0.05).
1
FCR = Feed conversion ratio (g dry feed consumed/g body weight gained) (Bailey and Aianärä, 2006).
2
FCE= Feed conversion efficiency (g wet weight gain/g dry diet fed) (De silva and Anderson, 1995).
3
SGR = Specific growth ratio (100[ln(final weight)×ln(initial weight)/day]) (López et la. 2006).
4
CF= Condition factor ((weight×105)/length3) (Tacon and Rodrigues, 1984).
498 A. Khodanazary et al. / Turk. J. Fish. Aquat. Sci. 13: 495-501 (2013)

findings reported by Lanari et al (1996), who found Another possible explanation for the effect of perlite
that additions of zeolite at 2.5 and 5% to rainbow on body weight gain is due to its effects on nutrient
trout diets improved weight gain and feed efficiency. absorption and as a carrier for aluminum silicate
Likewise, Yildirim et al (2009) indicated that fish fed (Scheideler, 1993).
on diets supplemented with zeolite at 1 and 2 % had
higher WG, SGR, protein efficiency rate and FCR Dry Matter and Protein Digestibility
than those fed diet without zeolite.
However, Dias et al (1998) reported that feeding The effects of zeolite and perlite addition on dry
European sea bass with diets containing 10 and 20% matter and protein digestibility coefficients are shown
natural zeolite as a bulk agent did not result in a in Table 3. Supplementing zeolite and perlite
remarkable effect on growth and feed utilization. increased digestibility coefficients of dry matter and
Absence of any significant effect of dietary zeolite on protein compared with the control. The highest
growth performance has also been reported for coho apparent digestibility coefficients of dietary dry
salmon and by Edsall and Smith (1989), red tilapia by matter and protein were achieved in the fish fed diets
Rafiee and Saad (2005) and Astacus leptodactylus by containing 5 % zeolite and perlite which are not
Zamani Kyasajmahaleh et al. (2007). different from each other (Table 3). Dias et al (1998)
Mumpton and Fishman (1977) noted that the observed that the dietary use of zeolite at 10 and 20%
growth enhancing effect of dietary zeolite may be had no significant effect on apparent digestibility
related to the type, properties and their incorporation coefficient of dietary protein. Also, Lanari et al
levels. The improved performance is likely associated (1996) investigated the effect of dietary zeolite in
with an improved nutrient utilization (Olver, 1989) rainbow trout, and found that dry matter and protein
and detoxifying effects of zeolite (Harvey et al. digestibility were not influenced by the incorporations
1993). The better nutrient utilization can be explained at 2.5 and 5%.
by a slower passage of pre-digested food through the
intestine, leading to the improved nutrients Bones and Scales Ash Content
absorption, particularly nitrogen (Dias et al., 1998;
Mumpton and Fishman, 1977). Ammonia is also For carp, significant (P<0.05) changes in ash
considered to act as a cell toxicant in higher animals, content of bones and scale were observed at the levels
and the prevention of its accumulation to toxic levels of 2.5 and 5% of either perlite or zeolite, compared to
in the intestinal tract could lead to a reduction of the control diet (Table 4). Similarly, Leach et al.
epithelial turnover, a sparing of energy and a better (1990) reported that using zeolite in the diet, had
nutrient utilization (Papaioannou et al., 2005). Ergün beneficial effects on bone ash and strength. Zolite and
et al. (2008) reported that the addition of clinoptilolite perlite have some ions which can be absorb by scales
(2.5 %) to the diet of young rainbow trout and bones.
significantly improved ammonia excretion rate.
The results of the present study reveal that Blood Serum Properties
supplementing perlite to fish diet could bring some
beneficial effects on growth. These results are in The mineral contents of serum from carps are
consistent with Glodek (1980), who stated that perlite shown in Table 5. Mg and P did not differ
used as a carrier in animal feed led to faster growth. significantly between the dietary treatments (P>0.05).

Table 3. Effect of zeolite and perlite on the apparent digestibility coefficients of dry matter and protein

Control Z 2.5 Z5 P 2.5 P5

ADC of dry matter (%)1 80.58±0.55d 83.00±0.89c 87.48±0.76a 84.87±0.71b 87.92±1.27a
ADC of protein (%)2 90.59±0.49d 91.99±0.28c 95.14±0.92a 93.42±0.64b 95.80±0.57a
Values are means ±S.D. of three replicates and values within the same row with different superscript letters are significantly different
(P<0.05).
1
ADC of dry matter (apparent digestibility coefficient of dry matter) = 100- 100 (%Cr in feed/%Cr in faeces) (Tibbetts et al., 2006).
2
ADC of protein (apparent digestibility coefficient of nutrient) = 100- 100 (%Cr in feed/%Cr in feces) (%nutrient in faeces/%nutrient in feed)
(Tibbetts et al., 2006).

Table 4. The effect of zeolite and perlite on ash contents of bones and scales

Control Z 2.5 Z5 P 2.5 P5

Ash of bones (%) 12.30±0.26e 13.73±0.22a 13.48±0.16b 12.95±0.58c 12.36±0.56d

Ash of scales (%) 4.05±0.46b 4.25±0.54a 3.72±0.18d 3.60±0.26e 3.97±0.58c
Values are means ±S.D. of three replicates and values within the same row with different superscript letters are significantly different
(P<0.05).
 A. Khodanazary et al. / Turk. J. Fish. Aquat. Sci. 13: 495-501 (2013) 499
Table 5. Serum mineral content of fish fed on perlite and zeolite

Control Z 2.5 Z5 P 2.5 P5

Na (mg/l) 159.33±2.88a 151.66±2.30a 140.33±9.07b 153.33±4.04a 156.00±8.88a
K (mg/l) 4.76±0.66bc 4.61±0.16c 5.70±0.41a 5.79±0.3a 5.39±0.3ab
Fe (mg/l) 109.66±12.66ab 131.66±10.78a 127.00±11.53a 80.33±13.01c 101.33±11.93bc
P (mg/l) 4.53±1.02a 4.00±0.10a 4.20±0.50a 4.00±0.98a 4.00±1.32a
Ca (mg/l) 9.10±0.10d 9.26±0.25d 10.90±0.00a 10.47±0.03b 10.03±0.05c
Mg (mg/l) 3.13±0.37a 2.93±0.45a 2.90±0.75a 2.26±0.30a 2.80±0.65a
Values are means ±S.D. of three replicates and values within the same row with different superscript letters are significantly different
(P<0.05).

Table 6. Serum protein, cholesterol and glucose content of fish fed on perlite and zeolite
Control Z 2.5 Z5 P 2.5 P5
Total protein (g/l) 2.76±0.66a 3.20±0.20a 3.13±0.05a 3.20±0.20a 2.93±0.20a
Cholesterol (mg/dl) 188.33±22.03a 169.66±27.15ab 156.66±10.11ab 131.33±34.93b 162.66±22.03ab
Glucose (mg/dl) 82.00±10.81b 101.00±26.21b 164.33±21.45a 100.66±18.87b 116.00±20.29b
Values are means ±S.D. of three replicates and values within the same row with different superscript letters are significantly different
(P<0.05).

Nevertheless, it was reported that the aluminum zeolite, except 20% increase in K level.
content of zeolite may from indigestible complexes Zeolite and perlite supplementation had not
with P, which could eventually reduce the availability significant effect on the serum total protein
of P to laying hens (Öztürk et al, 1998). Feeding low concentrations after 56 days of rearing (Table 6).
levels of dietary P is thought to reduce serum P that Cholesterol content of P 2.5 and glucose content of Z
this study which was found in the present study, 5 were significantly different from other treatments
where a decrease In P serum was noticed among (P<0.05). Wherease, Kanyilmaz, 2012 reported that
fishes fed on diets containing supplemental materials, blood glucose decreased with increasing of the zeolite
however, the differences was insignificant. According level in giltheah sea bream (Sparus aurata). The exact
to Table 5, Ca concentrations were no significantly mechanism of the effects of zeolite and perlite on the
different between control diet and Z 2.5. Ca cations in serum biochemical properties of fish is presently
zeolite are exchangeable with other cations such as unknown and we have no explanation for this
NH4+, Mg++, Na+, potassium (K+). Beneficial effects response.
may also be related to Al, Si or Na contents of zeolite
because these minerals were shown to influence Ca Conclusion
metabolism (Öztürk et al., 1998). However,
Shriatmadari (2008) reported that the decrease of P The results of the present work revealed that the
availability may be due to the increase of dietary Ca incorporations of zeolite or perlite into the diets of
utilization in zeolite. However, Enemark et al. (2003) common carp had positive effects on dry matter,
recorded initial slight decline in Ca concentrations in protein apparent digestibility coefficients, growth
serum which was likely caused by decreased performance. The optimum inclusion level of zeolite
availability of Ca from the zeolite containing diet. The and perlite appears to be 5 %. This research needs to
decrease in animal serum content of P and Mg was more study for different levels of zeolite and perlite
partly caused by interference of zeolite with intestinal and digestive enzymes in experimental fih
absorption and partly by a marginal dietary .
supplementation of these minerals (Trckova et al., Acknowledgements
2004).
Fish fed the Z 5 diet had the lowest Na value We wish to thank the staff of the Fishery
compared with the rest (P<0.05). Fish fed the control, Research Center, Gorgan University.
Z 2.5 and P 5 diet had significantly lower serum K
than those fed Z 5 and P 2.5. The serum Fe levels References
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