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Effect of Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of freshwater prawn Macrobrachium rosenbergii post larvae
The Journal of Basic & Applied Zoology (2014) xxx, xxx–xxx
The Egyptian German Society for Zoology
The Journal of Basic & Applied Zoology www.egsz.org www.sciencedirect.com
Effect of Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of freshwater prawn Macrobrachium rosenbergii post larvae C. Seenivasan *, S. Radhakrishnan, R. Shanthi, T. Muralisankar, P. Saravana Bhavan Crustacean Biology Laboratory, Department of Zoology, Bharathiar University, Coimbatore 641046, Tamil Nadu, India Received 29 June 2013; revised 19 December 2013; accepted 22 December 2013
KEYWORDS Prawn; L. sporogenes; Growth; Protein; Carbohydrate; Lipid
Abstract The present study was conducted to investigate the optimization of probiotic, Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of the freshwater prawn Macrobrachium rosenbergii post larvae (PL). Experimental diets were the same in all, except for the variation in probiotic levels. The probiotic L. sporogenes was used at 0%, 1%, 2%, 3% and 4% inclusion in the experimental diets. These diets were fed to M. rosenbergii PL for a period of 90 days. The food index parameters, such as SR, WG, SGR, FCE and PER were significantly (P < 0.05) higher in 4% L. sporogenes incorporated diet fed PL, whereas the FCR was significantly (P < 0.05) lower in 4% L. sporogenes incorporated diet fed PL. This indicates the fact that this feed produced higher growth rate than that of other experimental diets. Similarly the proximate composition of the total protein, total free amino acid, total carbohydrate, and total lipid content was significantly (P < 0.05) higher in 4% L. sporogenes incorporated diet fed PL. However, insignificant differences were recorded in ash and moisture contents between control and experimental groups. Energy utilization parameters, such as feeding rate, absorption rate, conversion rate and excretory rate were significantly (P < 0.05) higher in 4% L. sporogenes incorporated diet fed PL. Statistically insignificant differences were recorded in metabolic rate between control and experimental groups. This indicates that there were no differences in energy loss between control and experimental groups. However, L. sporogenes incorporated diet fed PL produced better growth performance. ª 2014 The Egyptian German Society for Zoology. Production and hosting by Elsevier B.V. All rights reserved.
* Corresponding author. Mobile: +91 9488175470. E-mail address: [email protected] (C. Seenivasan). Peer review under responsibility of The Egyptian German Society for Zoology.
Production and hosting by Elsevier
Introduction Macrobrachium rosenbergii is a species of aquaculture importance owing to its high fecundity, rapid growth, wide range of salinity and temperature tolerance, disease resistance as well as its superior taste and high commercial value (Johnson, 1982; New, 1995; Roustaian et al., 2001). M. rosenbergii is highly
2090-9896 ª 2014 The Egyptian German Society for Zoology. Production and hosting by Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.jobaz.2013.12.002
Please cite this article in press as: Seenivasan, C. et al., Effect of Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of freshwater prawn Macrobrachium rosenbergii post larvae (2014), http://dx.doi.org/10.1016/ j.jobaz.2013.12.002
2 preferred for culturing due to its fast growth rate, tolerance to wide fluctuations of temperature, salinity and resistance to major diseases. The use of probiotics in the aquatic organisms is increasing with the demand for more environment-friendly aquaculture practices (Gatesoupe, 1999). A probiotic is generally defined as a live microbial food supplement, which improves the balance of the host animal’s intestinal flora (Fuller, 1989). However in aquaculture, probiotics can be administered either as a food supplement or as an additive to water (Moriarty, 1998). Probiotics in aquaculture have been shown to have several modes of action, competitive exclusion of pathogenic bacteria through the production of inhibitory compounds, improvement of water quality, enhancement of immune response of host species and enhancement of nutrition of host species through the production of supplemental digestive enzymes (Gomez Gill et al., 2000; Verschuere et al., 2000). Probiotics have been shown to be effective in a wide range of species for the promotion of growth enhanced nutrition, immunity and survival rate. Venkat et al. (2004) has worked out different modes of administration of probiotics to M. roesenbrgii PL either through feed or as bioencapsulated in Artemia. Shinde et al. (2008) have reported the use of different commercial probiotic supplements administered through feeds to PL of M. rosenbergii. Saad et al. (2009) have reported the use of Biogen a probiotic for M. rosenbergii PL. However, the nutritional effects of probiotic bacteria, especially the effects of the bacteria on energy utilization have not been evaluated in aquaculture. Therefore, this study attempted to investigate the effect of probiotic, L. sporogenes on the survival, growth, biochemical constituents and energy utilization performance of M. rosenbergii PL. Materials and methods The post larvae of freshwater prawn, M. rosenbergii (PL 15) were purchased from a Happy Bay Annexe, Kanchipuram, Tamil Nadu, India and were stocked in a cement tank (1000 L) filled with freshwater. The PL were acclimated at ambient laboratory conditions for 15 days (up to PL 30) and starved for 24 h before the commencement of the feeding experiment. The experimental water had these physicochemical parameters: pH 7; total dissolved solids 0.90 g/L1; dissolved oxygen 7.20 mg/L1; BOD 30.00 mg/L1; COD 125.00 mg/ L1; ammonia 0.028 mg/L1.
C. Seenivasan et al. Mumbai, India) at 40 C until the pellets reached a constant weight and then stored in airtight jars at room temperature. The biochemical constituents of the experimental diets were determined, total protein content (Lowry et al., 1951), total free amino acids (Moore and Stein, 1948), total lipids (Folch et al., 1957), total carbohydrate (Roe, 1955), ash and moisture contents (APHA, 2005). These diets were freshly produced after 30 days to ensure high probiotic viability throughout the duration of feeding trail. In the control feed, no L. sporogenes was found throughout the duration of the feeding trail. Feeding experiment M. rosenbergii (PL-30) with the length and weight range of 1.34 ± 0.20 cm and 0.18 ± 0.04 g, respectively, were used for feeding experiment. Forty PL for each feed in triplicate were maintained in plastic tanks with 20/L water. The PL was maintained at the stocking density of 2/L. One group served as control, which was devoid of probiotics (0%). The experimental groups were fed with the respective concentration of L. sporogenes incorporated diets. The feeding was adjusted daily at 6:00 am and 6:00 pm. The daily ration was given at the rate of 10% of the body weight of PL with two equal halves, throughout the experimental period. The unfed feed, faeces and moult, if any, were collected after the respective hours of feeding. The feeding experiment was prolonged for 90 days; mild aeration was given continuously in order to maintain the optimal oxygen level. Determination of food indices After the feeding trial, food index parameters such as survival rate (SR), weight gain (WG), specific growth rate (SGR), feed conversion rate (FCR), feed conversion efficiency (FCE) and protein efficiency rate (PER), were individually determined by the following equations. Survival rateð%Þ ¼ Total No: of live animals= Total No: of initial animals 100 Weight gain ðgÞ ¼ Final weight ðgÞ Initial weight ðgÞ SGRð%Þ ¼ log w2 log w1 =t 100ðwhere;w1 &w2 ¼ Initial and final weights; respectively
Diet preparation
ðgÞ; and t ¼ Total number of experimental daysÞ
The composition of the experimental diets is given in Table 1. The probiotic L. sporogenes (Uni-Sankyo Ltd., Maharashtra, India) was incorporated into the test diets at five different concentrations individually 0% (control), 1%, 2%, 3% and 4%, respectively. Feed formulation was done basically by ‘‘Spearson’s square-method’’ using determined values of 40% protein content (Table 1). The proportion of each ingredient required was calculated precisely providing allowance for the premix. The dough was steam cooked and cooled to room temperature. After that, different concentrations of L. sporogenes were mixed with the dough and the feeds were pelletized separately with a locally made (Kolkata, India) hand pelletizer. The pellets were dried in a thermostatic oven (M/s Modern Industrial,
Condition factor ðCFÞ ¼ Prawn weight ðgÞ= Prawn length ðcmÞ 100 Feed conversion rate ðFCRÞ ¼ Total Feed intakeðgÞ=Total weight gain of the prawn ðgÞ
Feed conversion efficiency ðFCEÞ ¼ Biomass ðgÞ=Total Feed intake ðgÞ 100 Protein efficiency rate ðPERÞ ¼ Total Weight gain of PL ðgÞ=Total Protein consumed ðgÞ
Please cite this article in press as: Seenivasan, C. et al., Effect of Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of freshwater prawn Macrobrachium rosenbergii post larvae (2014), http://dx.doi.org/10.1016/ j.jobaz.2013.12.002
L. sporogenes on growth promotion of prawn Table 1
3
Ingredients and proximate composition of prepared diets.
Ingredients (%)
Control
Experiment (L. sporogenes incorporated) 1%
2%
3%
4%
Fish meal Ground nut oil cake Soybean meal Corn flour Egg albumin Topica flour Cod liver oil Vitamin B-complex mix L. sporogenes Total
33.84 25.00 24.00 4.00 5.06 5.10 2.00 1.00 0 100
33.84 25.00 24.00 3.00 5.06 5.10 2.00 1.00 1.00 100
33.84 25.00 23.00 3.00 5.06 5.10 2.00 1.00 2.00 100
34.84 25.00 21.00 3.00 5.06 5.10 2.00 1.00 3.00 100
35.84 24.00 20.00 3.00 5.06 5.10 2.00 1.00 4.00 100
Proximate composition Protein (%) Carbohydrate (%) Lipid (%) Ash (%) Moisture (%) Digestible energy (k cal/kg)
40.10 21.76 9.28 14.00 9.50 3296.86
40.00 21.10 9.24 13.00 9.90 3262.52
39.63 20.71 9.17 12.00 9.40 3228.17
39.52 20.01 9.08 13.00 9.10 3193.83
39.40 19.50 8.90 14.00 9.10 3159.49
Bioenergetics
Biochemical constituents of the experimental animals
The energy content of the whole prawn, feeds, moult and faeces was measured using Parr 1281 Oxygen Bomb Calorimeter. The energy budget was calculated using the equation (C = (P + E) + R + F + U) derived by Petrusewicz and Macfadyen (1970); where, C is the energy consumed in food; P is the growth; R is the material lost as heat due to metabolism; F is the energy lost in faeces; U is the energy lost in excretion and; E is the energy lost in exuviated.
In the initial and final days of the experiment, the biochemical constituents of the experimental animals were determined. The biochemical constituents, such as total protein content (Lowry et al., 1951), total free amino acid (Moore and Stein, 1948), total lipids (Folch et al., 1957), total carbohydrate (Roe, 1955), ash and moisture contents (APHA, 2005) of individual diet fed prawns were measured.
Mean Food Consumption ðk cal=ðdayÞ
Statistical analyses
Feeding Rate ðFRÞ ¼ Initial live weight of the prawn ðgÞ
A one-way analysis of variance (ANOVA; SPSS, 13.0) was used to determine whether significant variation between the treatments existed. Differences between means were determined and compared by the post hoc multiple comparison test (DMRT). All tests used a significance level of P < 0.05. Data are reported as mean ± standard deviation.
Mean Absorption ¼ Mean Food Consumptionðk cal=dayÞ Mean Food discharged as Faecesðk cal=dayÞ
Mean Absorptionðk cal=dayÞ Absorption Rate ðARÞ ¼ Initial live weight of the prawnðgÞ Mean Conversion ¼ Mean weight gainðk cal=dayÞ þ Mean exuvial weightðk cal=dayÞ
Results and discussion Biochemical constituents of experimental diet
MeanNH3 Excretionðk cal=dayÞNH3
Biochemical constituents of L. sporogenes incorporated diets are presented in Table 1. The level of total protein and total lipids was found to be 40.10–39.40% and 9.28–8.90%, respectively. The levels of ash and moisture contents were 12.00– 14.00% and 9.10–9.90%, respectively. The digestible energy of these diets ranged from 3228.17 to 3296.86 k cal/kg1 (Table 1).
NH3 Excretion RateðERÞ ¼ Initial live weight of the prawnðgÞ
Biochemical constituents of experimental animals
Mean Conversionðk cal=dayÞ Conversion rateðCRÞ ¼ Initial live weight of the prawnðgÞ
Metabolic RateðMRÞ ¼ Absorption Rateðk cal=g=dayÞ Conversion Rateðk cal=g=dayÞ þ NH3 Excretion Rateðk cal=g=dayÞ
The initial body concentration of biochemical constituents, such as total protein content, free amino acids, total carbohydrate, total lipids, ash and moisture contents of PL was recorded to be 28%, 11%, 6.5%, 3%, 8.80% and 80%,
Please cite this article in press as: Seenivasan, C. et al., Effect of Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of freshwater prawn Macrobrachium rosenbergii post larvae (2014), http://dx.doi.org/10.1016/ j.jobaz.2013.12.002
4
C. Seenivasan et al. Table 2
Proximate composition of M. rosenbergii PL fed with L. sporogenes supplemented diet.
Treatments
Protein content (%)
Free amino acid (%)
Total carbohydrate (%)
Total lipids (%)
Ash (%)
Moisture (%)
Initial Control 1% LS 2% LS 3% LS 4% LS F-value
28.00 ± 2.33 57.60 ± 2.60d 59.00 ± 2.30cd 60.00 ± 2.14bc 62.00 ± 2.34ab 63.40 ± 2.02a 12.80
11.00 ± 1.08 24.00 ± 1.33d 26.40 ± 1.28cd 28.00 ± 1.64bc 30.40 ± 1.88ab 32.10 ± 1.54a 12.80
6.50 ± 1.01 10.30 ± 1.06c 12.90 ± 1.05b 14.50 ± 1.09b 17.56 ± 1.12a 19.18 ± 1.03a 33.19
3.12 ± 0.88 6.80 ± 1.04c 7.60 ± 1.00bc 8.20 ± 1.07bc 9.00 ± 1.03ab 11.00 ± 1.01a 7.06
8.80 ± 0.50 17.02 ± 1.66a 18.20 ± 1.78a 18.60 ± 2.00a 18.40 ± 1.71a 19.00 ± 1.05a <1
80.00 ± 4.00 76.10 ± 3.20a 75.60 ± 2.90a 75.30 ± 2.50a 75.10 ± 3.00a 75.00 ± 2.70a <1
Each value is a mean ± SD of three replicate analyses, within each row means with different superscript letters are statistically significant P < 0.05 (one way ANOVA and subsequently post hoc multiple comparison with DMRT).
respectively (Table 2). The proximate composition of M. rosenbergii offered diets with various levels of L. sporogenes is given in Table 2. There was a decrease (P < 0.05) in the moisture content while the protein, amino acid, carbohydrate, lipid and ash contents increased (P < 0.05) in all the experimental groups. The highest increase (P < 0.05) in total protein, free amino acid, total carbohydrate, total lipids and ash contents were recorded in the 4% L. sporogenes incorporated diet, followed by 3%, 2% and 1% and the lowest increase was recorded in the control. These differences in proximate composition were statistically significant (P < 0.05), except the ash. It has been reported that in M. rosenbergii PL fed with B. subtilis incorporated diets, there was significant improvement in the proximate composition of body tissues (Seenivasan et al., 2012b). Seenivasan et al., 2011 have reported that with the commercial probiotic, Binifit supplemented diets; there was significant improvement in the proximate composition of body tissues in M. rosenbergii PL. The beneficial effects of probiotics in M. rosenbergii PL culture have also been supported by Venkat et al. (2004), Suralikar and Sahu (2001) and Shinde et al. (2008). Survival performance Under the four levels of L. sporogenes supplement, among the experimental groups, prawns exhibited normal behaviour and no cannibalism was observed. Therefore, L. sporogenes incorporated feed was accepted by the prawns at all levels and showed a significantly higher survival rate. The survival rate was significantly (P < 0.05) higher in M. rosenbergii PL fed with 4% L. sporogenes incorporated diets when compared with other experimental groups (Table 3). Similar improved survival has been reported in M. rosenbergii PL fed with Biogen supplemented diets (Saad et al., 2009) and diets supplemented
Table 3
with different probiotics (Shinde et al., 2008) and in the Indian white shrimp, Fenneropenaus indicus after feeding with Bacillus (Ziaei-Nejad et al., 2006). In general, L. sporogenes has the ability to attach with the gut epithelium and establish there. In the present study, the higher survival rate observed in L. sporogenes incorporated diet fed PL may be due to the administration of significant changes in the proportion of the population of the gut micro flora. This may be exerted by the elimination of harmful bacteria due to establishment of the beneficial probionts, L. sporogenes in the intestine of M. rosenbergii. Or, colonization of L. sporogenes may be dominant over harmful bacteria. By their large presence, saturate the adhesion receptors and prevent the pathogenic bacteria from attachment and colonization (Vine et al., 2004; Seenivasan et al., 2012a). Growth performance L. sporogenes supplement diet appeared to provide beneficial effects on the growth of M. rosenbergii PL. The results indicated that the prawn which had access to diet containing L. sporogenes, exhibited greater growth than that of the control diet (Table 3). There were significant differences in growth increment (P < 0.05) between different experimental and control groups. Final body weight gain, specific growth rate and food conversion ratio in PL offered with L. sporogenes incorporated diets, particularly at 4%, were superior to those of other experimental groups as well as control. Thus, inclusion of L. sporogenes >1% is recommended. Similar results on significant improvement in growth, weight gain and SGR in M. rosenbergii fed with bio-encapsulated L. sporogenes were reported by Seenivasan et al. (2012a). It has been reported that the growth performance, SGR and weight gain were higher in L. sporogenes, B. subtilis and S. cerevisiae supplemented diet
Food indices of M. rosenbergii PL fed with L. sporogenes supplemented diet.
Parameters Survival rate (%) Weight gain (g) Specific growth rate (%) Condition factor (%) Food conversion ratio (g) Food conversion efficiency (%) Protein efficiency rate (g)
Control
1% LS c
77.50 ± 2.50 0.82 ± 0.06c 0.827 ± 0.30c 1.94 ± 0.20a 3.40 ± 0.28a 0.859 ± 0.06b 0.642 ± 0.13b
2% LS c
80.00 ± 2.50 0.85 ± 0.02c 0.841 ± 0.028c 1.73 ± 0.14ab 2.93 ± 0.22ab 0.850 ± 0.08b 0.755 ± 0.11ab
3% LS b
85.00 ± 2.50 0.90 ± 0.04c 0.864 ± 0.029bc 1.63 ± 0.18ab 2.77 ± 0.33ab 0.877 ± 0.10b 0.801 ± 0.09ab
4% LS ab
87.50 ± 2.50 1.01 ± 0.07b 0.911 ± 0.036b 1.45 ± 0.10b 2.55 ± 0.48ab 0.959 ± 0.05b 0.868 ± 0.07a
F-value a
90.00 ± 2.50 1.22 ± 0.09a 0.989 ± 0.039a 1.57 ± 0.22b 2.36 ± 0.80a 1.120 ± 0.12a 0.939 ± 0.10a
12.90 38.65 12.09 3.40 2.15 5.20 3.67
Each value is a mean ± SD of three replicate analyses, within each row means with different superscript letters are statistically significant P < 0.05 (one way ANOVA and subsequently post hoc multiple comparison with DMRT).
Please cite this article in press as: Seenivasan, C. et al., Effect of Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of freshwater prawn Macrobrachium rosenbergii post larvae (2014), http://dx.doi.org/10.1016/ j.jobaz.2013.12.002
L. sporogenes on growth promotion of prawn Table 4
5
Energy budget of M. rosenbergii PL fed with L. sporogenes supplemented diet (values are expressed as k cal/g/day).
Parameters
Control
1% LS
2% LS
3% LS
4% LS
F-value
Feeding rate Absorption rate Conversion rate NH3 Excretory rate Metabolic rate
0.466 ± 0.029c 0.402 ± 0.033c 0.220 ± 0.022b 0.009 ± 0.002b 0.194 ± 0.014b
0.480 ± 0.0320c 0.429 ± 0.030c 0.232 ± 0.027b 0.010 ± 0.003b 0.210 ± 0.017ab
0.511 ± 0.027bc 0.465 ± 0.028bc 0.255 ± 0.031b 0.012 ± 0.002b 0.224 ± 0.019ab
0.549 ± 0.026b 0.508 ± 0.048ab 0.286 ± 0.026ab 0.015 ± 0.003ab 0.237 ± 0.026a
0.605 ± 0.031a 0.568 ± 0.0.41a 0.345 ± 0.034a 0.016 ± 0.001a 0.239 ± 0.021a
11.51 9.59 5.56 4.63 2.75
Each value is a mean ± SD of three replicate analyses, within each row means with different superscript letters are statistically significant P < 0.05 (one way ANOVA and subsequently post hoc multiple comparison with DMRT).
fed M. rosenbergii PL (Seenivasan et al., 2012c). It has been also reported that the growth, SGR and weight gain were increased in bioencapsulated Lactobacillus spp. fed fish, Carassius auratus and Xiphophorus helleri (Ashim et al., 2009). According to Shinde et al. (2008), different probiotics supplemented diets had improved growth rate, weight gain, FCR and FER of M. rosenbergii PL. Therefore, in the present study, improvement of growth and food conversion may be due to L. sporogenes. In addition to growth, the feed conversion efficiency, protein efficiency rate and condition factor were also significantly better (P < 0.05) in PL offered diets with L. sporogenes when compared with control. A possible explanation for the increased performance as a result of the addition of the probiotic is the improvement in digestibility, which may in turn explain the observed better growth rate and the efficiency of feed conversion. Otherwise, probiotics influence digestive processes by enhancing microbial enzyme activity, and consequently improving the digestion, absorption and utilization of the feed (Bomba et al., 2002). Similar observations have been reported by De Schrijver and Ollevier (2000) in turbot, S. maximus fed with the bacteria, Vibrio proteolyticus incorporated diet and by Lara-Flores et al. (2003) in tilapia fry fed with 0.1% probiotics incorporated diets, which improved animal growth and apparent protein digestibility. The higher PER recorded in the present study indicates that the diets with probiotics have significantly improved dietary protein and energy utilization by M. rosenbergii. Venkat et al. (2004) showed a similar improvement in the PER in M. rosenbergii fed probiotics supplemented diets; Bagheri et al. (2008) observed significant improvements of PER in rainbow trout fry (1 g) fed with administration of B. licheniformis + B. subtilis at levels P log 9 cells g1. In the present study, the results indicated that the L. sporogenes incorporation showed positive effect of growth promotion in M. rosenbergii PL. Therefore, the overall growth was higher particularly in 4% L. sporogenes incorporated diet fed M. rosenbergii. Energy utilization performance In the present study, the energy budget of M. rosenbergii offered diets with various levels of L. sporogenes inclusions is given in Table 4. There were significant increases in the feeding rate, absorption rate, conversion rate and NH3 execratory rate in the experimental groups, when compared with the control (P < 0.05). The observed increment was higher at 4% L. sporogenes incorporated diet followed by 3%, 2%, and 1%, and the lowest value was recorded in the control group. It has been reported that the growth rate and feed energy utilization were increased in Saccharomyces cerevisiae supplemented diet fed M. rosenbergii PL (Seenivasan et al., 2013). It has also been
reported that the growth rate performance and nutrient energy utilization were higher in L. sporogenes, B. subtilis and S. cerevisiae inclusion diet fed M. rosenbergii PL (Seenivasan et al., 2012b–e). It has also been reported that the growth rate performance and nutrient energy utilization were higher in Binifit inclusion pellet fed M. rosenbergii PL (Seenivasan et al., 2011). Dhanaraja et al. (2010) reported that probiotics L. acidophilus and yeast S. cerevisiae supplemented diets had improved the energy budget of Koi Carp, C. carpio. It has been reported that the growth rate and feed energy utilization were increased in S. cerevisiae supplemented diet fed Nile tilapia, O. niloticus (Abdel-Tawwab et al., 2008). It has also been reported that the growth rate performance and nutrient energy utilization were higher in Biogen inclusion diet fed Nile tilapia (Haroun et al., 2006). Immanuel et al. (2003) reported that the probiotics Lactobacillus and yeast supplemented diets had improved the energy utilization of pearl spot Etroplus suratensis. From the above discussion, it may be concluded that L. sporogenes incorporation has significantly improved survival, growth, SGR, FCR, PER, FCE, biochemical constituents and energy utilization in post larvae of M. rosenbergii. Therefore, L. sporogenes can be supplemented in formulated diets for the maintenance of healthy M. rosenbergii at nursery and grows out in ponds. Also, M. rosenbergii can be used in farm management at a small scale level, and consequently, inland aquaculture of Macrobrachium may be promoted in a sustainable manner.
Acknowledgement The Bharathiar University, Coimbatore, Tamil Nadu, India is gratefully acknowledged for the financial support provided in the form of University Research Fellowship to Mr. C. Seenivasan. We thank Dr. G. Immanuel, Assistant Professor, Centre for Marine Sciences and Technology, Manonmaniam Sundaranar University, Tamil Nadu, India for providing his subject expertise to conduct the experiment. References Abdel-Tawwab, M., Abdel-Rahman, A.M., Ismael, N.E.M., 2008. Evaluation of commercial live baker’s yeast, Saccharomyces cerevisiae as a growth and immunity promoter for fry Nile tilapia, Oreochromis niloticus (L.) challenged in situ with Aeromonas hydrophila. Aquaculture 280, 185–189. APHA, 2005. Standard Methods for the Examination of Water and Wastewater, 20th ed. American Public Health Association, New York.
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C. Seenivasan et al. Roustaian, P., Kamarudin, M.S., Omar, H.B., Saad, C.R., Ahmad, M.H., 2001. Biochemical changes in freshwater prawn Macrobrachium rosenbergii during larval development. J. World Soc. Aquacult. 32, 52–59. Saad, S.A., Habashy, M.M., Sharshar, M.K., 2009. Growth response of the freshwater prawn, Macrobrachium rosenbergii (De Man), to diets having different levels of Biogen. World Appl. Sci. J. 6, 550–556. Seenivasan, C., Saravana Bhavan, P., Radhakrishnan, S., 2011. Effect of probiotics (BinifitTM) on survival, growth, biochemical constituents and energy budget of the freshwater prawn Macrobrachium rosenbergii post larvae. Elixir Aquacult. 41, 5919–5927. Seenivasan, C., Saravana Bhavan, P., Radhakrishnan, S., Shanthi, R., 2012a. Enrichment of Artemia nauplii with Lactobacillus sporogenes for enhancing the survival, growth and levels of biochemical of constituents in the post-larvae of freshwater prawn Macrobrachium rosenbergii. Turkish J. Fish. Aquatic Sci. 12, 23–31. Seenivasan, C., Radhakrishnan, S., Muralisankar, T., Saravana Bhavan, P., 2012b. Influence of combined probiotics Lactobacillus sporogenes and Bacillus subtilis on survival, growth, biochemical changes and energy utilization performance of Macrobrachium rosenbergrii (De Man 1879) post larvae. J. Ecobiotechnol. 4, 29–34. Seenivasan, C., Saravana Bhavan, P., Radhakrishnan, S., Muralisankar, T., 2012c. Effects of probiotics on survival, growth and biochemical constituents of freshwater prawn Macrobrachium rosenbergii post larvae. Turkish J. Fish. Aquatic Sci. 12, 331–338. Seenivasan, C., Radhakrishnan, S., Muralisankar, T., Saravana Bhavan, P., 2012d. Efficacy of probiotics on survival, growth, biochemical changes and energy utilization performance of Macrobrachium rosenbergii (De Man 1879) post larvae. J. Sci. Res. 4, 729–740. Seenivasan, C., Radhakrishnan, S., Muralisankar, T., Saravana Bhavan, P., 2012e. Bacillus subtilis on survival, growth, biochemical constituents and energy utilization of the freshwater prawn Macrobrachium rosenbergii post larvae. Egyptian J. Aquatic Res. 38, 195–203. Seenivasan, C., Saravana Bhavan, P., Radhakrishnan, S., Muralisankar, T., Srinivasan, V., Manickam, N., 2013. Effect of Saccharomyces cerevisiae on survival, growth, biochemical constituents and energy utilization in the prawn Macrobrachium rosenbergii. Int. J. Appl. Biol. Pharmaceutical Technol. 4, 39–47. Shinde, A.N., Mulye, V.B., Chogale, N.D., Bhatkar, V.R., Bondre, R.D., Mohite, A.S., 2008. Effect of different probiotics Macrobrachium Rosenbergii (De-Man) post larvae. Aquaculture 9, 7–12. Suralikar, V., Sahu, N.P., 2001. Effect of feeding probiotic (Lactobacillus cremoris) on growth and survival of Macrobrachium rosenbergii postlarvae. J. Appl. Animal Res. 20, 117–124. Venkat, H.K., Narottam, P.Sahu., Kamal, K.Jain., 2004. Effect of feeding Lactobacillus based probiotics on the gut microflora, growth and survival of postlarvae of Macrobrachium rosenbergii (De Man). Aquac. Res. 35, 501–507. Verschuere, L., Rombaut, G., Sorgeloos, P., Verstraete, W., 2000. Probiotic bacteria as biological control agents in aquaculture. Microbiol. Mol. Biol. 64, 655–671. Vine, N.G., Leukes, W.D., Kaiser, H., 2004. In vitro growth characteristics of five candidate aquaculture probiotics and two fish pathogens grown in fish intestinal mucus. FEMS Microbiol. Lett. 231, 145–152. Ziaei-Nejad, S., Rezaei, M.H., Takami, G.A., Lovett, D.L., Mirvaghefi, A.R., Shakouri, M., 2006. The effect of Bacillus spp. bacteria used as probiotics on digestive enzyme activity, survival and growth in the Indian white shrimp Fenneropenaeus indicus. Aquaculture 252, 516–524.
Please cite this article in press as: Seenivasan, C. et al., Effect of Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of freshwater prawn Macrobrachium rosenbergii post larvae (2014), http://dx.doi.org/10.1016/ j.jobaz.2013.12.002
The Egyptian German Society for Zoology
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Effect of Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of freshwater prawn Macrobrachium rosenbergii post larvae C. Seenivasan *, S. Radhakrishnan, R. Shanthi, T. Muralisankar, P. Saravana Bhavan Crustacean Biology Laboratory, Department of Zoology, Bharathiar University, Coimbatore 641046, Tamil Nadu, India Received 29 June 2013; revised 19 December 2013; accepted 22 December 2013
KEYWORDS Prawn; L. sporogenes; Growth; Protein; Carbohydrate; Lipid
Abstract The present study was conducted to investigate the optimization of probiotic, Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of the freshwater prawn Macrobrachium rosenbergii post larvae (PL). Experimental diets were the same in all, except for the variation in probiotic levels. The probiotic L. sporogenes was used at 0%, 1%, 2%, 3% and 4% inclusion in the experimental diets. These diets were fed to M. rosenbergii PL for a period of 90 days. The food index parameters, such as SR, WG, SGR, FCE and PER were significantly (P < 0.05) higher in 4% L. sporogenes incorporated diet fed PL, whereas the FCR was significantly (P < 0.05) lower in 4% L. sporogenes incorporated diet fed PL. This indicates the fact that this feed produced higher growth rate than that of other experimental diets. Similarly the proximate composition of the total protein, total free amino acid, total carbohydrate, and total lipid content was significantly (P < 0.05) higher in 4% L. sporogenes incorporated diet fed PL. However, insignificant differences were recorded in ash and moisture contents between control and experimental groups. Energy utilization parameters, such as feeding rate, absorption rate, conversion rate and excretory rate were significantly (P < 0.05) higher in 4% L. sporogenes incorporated diet fed PL. Statistically insignificant differences were recorded in metabolic rate between control and experimental groups. This indicates that there were no differences in energy loss between control and experimental groups. However, L. sporogenes incorporated diet fed PL produced better growth performance. ª 2014 The Egyptian German Society for Zoology. Production and hosting by Elsevier B.V. All rights reserved.
* Corresponding author. Mobile: +91 9488175470. E-mail address: [email protected] (C. Seenivasan). Peer review under responsibility of The Egyptian German Society for Zoology.
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Introduction Macrobrachium rosenbergii is a species of aquaculture importance owing to its high fecundity, rapid growth, wide range of salinity and temperature tolerance, disease resistance as well as its superior taste and high commercial value (Johnson, 1982; New, 1995; Roustaian et al., 2001). M. rosenbergii is highly
2090-9896 ª 2014 The Egyptian German Society for Zoology. Production and hosting by Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.jobaz.2013.12.002
Please cite this article in press as: Seenivasan, C. et al., Effect of Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of freshwater prawn Macrobrachium rosenbergii post larvae (2014), http://dx.doi.org/10.1016/ j.jobaz.2013.12.002
2 preferred for culturing due to its fast growth rate, tolerance to wide fluctuations of temperature, salinity and resistance to major diseases. The use of probiotics in the aquatic organisms is increasing with the demand for more environment-friendly aquaculture practices (Gatesoupe, 1999). A probiotic is generally defined as a live microbial food supplement, which improves the balance of the host animal’s intestinal flora (Fuller, 1989). However in aquaculture, probiotics can be administered either as a food supplement or as an additive to water (Moriarty, 1998). Probiotics in aquaculture have been shown to have several modes of action, competitive exclusion of pathogenic bacteria through the production of inhibitory compounds, improvement of water quality, enhancement of immune response of host species and enhancement of nutrition of host species through the production of supplemental digestive enzymes (Gomez Gill et al., 2000; Verschuere et al., 2000). Probiotics have been shown to be effective in a wide range of species for the promotion of growth enhanced nutrition, immunity and survival rate. Venkat et al. (2004) has worked out different modes of administration of probiotics to M. roesenbrgii PL either through feed or as bioencapsulated in Artemia. Shinde et al. (2008) have reported the use of different commercial probiotic supplements administered through feeds to PL of M. rosenbergii. Saad et al. (2009) have reported the use of Biogen a probiotic for M. rosenbergii PL. However, the nutritional effects of probiotic bacteria, especially the effects of the bacteria on energy utilization have not been evaluated in aquaculture. Therefore, this study attempted to investigate the effect of probiotic, L. sporogenes on the survival, growth, biochemical constituents and energy utilization performance of M. rosenbergii PL. Materials and methods The post larvae of freshwater prawn, M. rosenbergii (PL 15) were purchased from a Happy Bay Annexe, Kanchipuram, Tamil Nadu, India and were stocked in a cement tank (1000 L) filled with freshwater. The PL were acclimated at ambient laboratory conditions for 15 days (up to PL 30) and starved for 24 h before the commencement of the feeding experiment. The experimental water had these physicochemical parameters: pH 7; total dissolved solids 0.90 g/L1; dissolved oxygen 7.20 mg/L1; BOD 30.00 mg/L1; COD 125.00 mg/ L1; ammonia 0.028 mg/L1.
C. Seenivasan et al. Mumbai, India) at 40 C until the pellets reached a constant weight and then stored in airtight jars at room temperature. The biochemical constituents of the experimental diets were determined, total protein content (Lowry et al., 1951), total free amino acids (Moore and Stein, 1948), total lipids (Folch et al., 1957), total carbohydrate (Roe, 1955), ash and moisture contents (APHA, 2005). These diets were freshly produced after 30 days to ensure high probiotic viability throughout the duration of feeding trail. In the control feed, no L. sporogenes was found throughout the duration of the feeding trail. Feeding experiment M. rosenbergii (PL-30) with the length and weight range of 1.34 ± 0.20 cm and 0.18 ± 0.04 g, respectively, were used for feeding experiment. Forty PL for each feed in triplicate were maintained in plastic tanks with 20/L water. The PL was maintained at the stocking density of 2/L. One group served as control, which was devoid of probiotics (0%). The experimental groups were fed with the respective concentration of L. sporogenes incorporated diets. The feeding was adjusted daily at 6:00 am and 6:00 pm. The daily ration was given at the rate of 10% of the body weight of PL with two equal halves, throughout the experimental period. The unfed feed, faeces and moult, if any, were collected after the respective hours of feeding. The feeding experiment was prolonged for 90 days; mild aeration was given continuously in order to maintain the optimal oxygen level. Determination of food indices After the feeding trial, food index parameters such as survival rate (SR), weight gain (WG), specific growth rate (SGR), feed conversion rate (FCR), feed conversion efficiency (FCE) and protein efficiency rate (PER), were individually determined by the following equations. Survival rateð%Þ ¼ Total No: of live animals= Total No: of initial animals 100 Weight gain ðgÞ ¼ Final weight ðgÞ Initial weight ðgÞ SGRð%Þ ¼ log w2 log w1 =t 100ðwhere;w1 &w2 ¼ Initial and final weights; respectively
Diet preparation
ðgÞ; and t ¼ Total number of experimental daysÞ
The composition of the experimental diets is given in Table 1. The probiotic L. sporogenes (Uni-Sankyo Ltd., Maharashtra, India) was incorporated into the test diets at five different concentrations individually 0% (control), 1%, 2%, 3% and 4%, respectively. Feed formulation was done basically by ‘‘Spearson’s square-method’’ using determined values of 40% protein content (Table 1). The proportion of each ingredient required was calculated precisely providing allowance for the premix. The dough was steam cooked and cooled to room temperature. After that, different concentrations of L. sporogenes were mixed with the dough and the feeds were pelletized separately with a locally made (Kolkata, India) hand pelletizer. The pellets were dried in a thermostatic oven (M/s Modern Industrial,
Condition factor ðCFÞ ¼ Prawn weight ðgÞ= Prawn length ðcmÞ 100 Feed conversion rate ðFCRÞ ¼ Total Feed intakeðgÞ=Total weight gain of the prawn ðgÞ
Feed conversion efficiency ðFCEÞ ¼ Biomass ðgÞ=Total Feed intake ðgÞ 100 Protein efficiency rate ðPERÞ ¼ Total Weight gain of PL ðgÞ=Total Protein consumed ðgÞ
Please cite this article in press as: Seenivasan, C. et al., Effect of Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of freshwater prawn Macrobrachium rosenbergii post larvae (2014), http://dx.doi.org/10.1016/ j.jobaz.2013.12.002
L. sporogenes on growth promotion of prawn Table 1
3
Ingredients and proximate composition of prepared diets.
Ingredients (%)
Control
Experiment (L. sporogenes incorporated) 1%
2%
3%
4%
Fish meal Ground nut oil cake Soybean meal Corn flour Egg albumin Topica flour Cod liver oil Vitamin B-complex mix L. sporogenes Total
33.84 25.00 24.00 4.00 5.06 5.10 2.00 1.00 0 100
33.84 25.00 24.00 3.00 5.06 5.10 2.00 1.00 1.00 100
33.84 25.00 23.00 3.00 5.06 5.10 2.00 1.00 2.00 100
34.84 25.00 21.00 3.00 5.06 5.10 2.00 1.00 3.00 100
35.84 24.00 20.00 3.00 5.06 5.10 2.00 1.00 4.00 100
Proximate composition Protein (%) Carbohydrate (%) Lipid (%) Ash (%) Moisture (%) Digestible energy (k cal/kg)
40.10 21.76 9.28 14.00 9.50 3296.86
40.00 21.10 9.24 13.00 9.90 3262.52
39.63 20.71 9.17 12.00 9.40 3228.17
39.52 20.01 9.08 13.00 9.10 3193.83
39.40 19.50 8.90 14.00 9.10 3159.49
Bioenergetics
Biochemical constituents of the experimental animals
The energy content of the whole prawn, feeds, moult and faeces was measured using Parr 1281 Oxygen Bomb Calorimeter. The energy budget was calculated using the equation (C = (P + E) + R + F + U) derived by Petrusewicz and Macfadyen (1970); where, C is the energy consumed in food; P is the growth; R is the material lost as heat due to metabolism; F is the energy lost in faeces; U is the energy lost in excretion and; E is the energy lost in exuviated.
In the initial and final days of the experiment, the biochemical constituents of the experimental animals were determined. The biochemical constituents, such as total protein content (Lowry et al., 1951), total free amino acid (Moore and Stein, 1948), total lipids (Folch et al., 1957), total carbohydrate (Roe, 1955), ash and moisture contents (APHA, 2005) of individual diet fed prawns were measured.
Mean Food Consumption ðk cal=ðdayÞ
Statistical analyses
Feeding Rate ðFRÞ ¼ Initial live weight of the prawn ðgÞ
A one-way analysis of variance (ANOVA; SPSS, 13.0) was used to determine whether significant variation between the treatments existed. Differences between means were determined and compared by the post hoc multiple comparison test (DMRT). All tests used a significance level of P < 0.05. Data are reported as mean ± standard deviation.
Mean Absorption ¼ Mean Food Consumptionðk cal=dayÞ Mean Food discharged as Faecesðk cal=dayÞ
Mean Absorptionðk cal=dayÞ Absorption Rate ðARÞ ¼ Initial live weight of the prawnðgÞ Mean Conversion ¼ Mean weight gainðk cal=dayÞ þ Mean exuvial weightðk cal=dayÞ
Results and discussion Biochemical constituents of experimental diet
MeanNH3 Excretionðk cal=dayÞNH3
Biochemical constituents of L. sporogenes incorporated diets are presented in Table 1. The level of total protein and total lipids was found to be 40.10–39.40% and 9.28–8.90%, respectively. The levels of ash and moisture contents were 12.00– 14.00% and 9.10–9.90%, respectively. The digestible energy of these diets ranged from 3228.17 to 3296.86 k cal/kg1 (Table 1).
NH3 Excretion RateðERÞ ¼ Initial live weight of the prawnðgÞ
Biochemical constituents of experimental animals
Mean Conversionðk cal=dayÞ Conversion rateðCRÞ ¼ Initial live weight of the prawnðgÞ
Metabolic RateðMRÞ ¼ Absorption Rateðk cal=g=dayÞ Conversion Rateðk cal=g=dayÞ þ NH3 Excretion Rateðk cal=g=dayÞ
The initial body concentration of biochemical constituents, such as total protein content, free amino acids, total carbohydrate, total lipids, ash and moisture contents of PL was recorded to be 28%, 11%, 6.5%, 3%, 8.80% and 80%,
Please cite this article in press as: Seenivasan, C. et al., Effect of Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of freshwater prawn Macrobrachium rosenbergii post larvae (2014), http://dx.doi.org/10.1016/ j.jobaz.2013.12.002
4
C. Seenivasan et al. Table 2
Proximate composition of M. rosenbergii PL fed with L. sporogenes supplemented diet.
Treatments
Protein content (%)
Free amino acid (%)
Total carbohydrate (%)
Total lipids (%)
Ash (%)
Moisture (%)
Initial Control 1% LS 2% LS 3% LS 4% LS F-value
28.00 ± 2.33 57.60 ± 2.60d 59.00 ± 2.30cd 60.00 ± 2.14bc 62.00 ± 2.34ab 63.40 ± 2.02a 12.80
11.00 ± 1.08 24.00 ± 1.33d 26.40 ± 1.28cd 28.00 ± 1.64bc 30.40 ± 1.88ab 32.10 ± 1.54a 12.80
6.50 ± 1.01 10.30 ± 1.06c 12.90 ± 1.05b 14.50 ± 1.09b 17.56 ± 1.12a 19.18 ± 1.03a 33.19
3.12 ± 0.88 6.80 ± 1.04c 7.60 ± 1.00bc 8.20 ± 1.07bc 9.00 ± 1.03ab 11.00 ± 1.01a 7.06
8.80 ± 0.50 17.02 ± 1.66a 18.20 ± 1.78a 18.60 ± 2.00a 18.40 ± 1.71a 19.00 ± 1.05a <1
80.00 ± 4.00 76.10 ± 3.20a 75.60 ± 2.90a 75.30 ± 2.50a 75.10 ± 3.00a 75.00 ± 2.70a <1
Each value is a mean ± SD of three replicate analyses, within each row means with different superscript letters are statistically significant P < 0.05 (one way ANOVA and subsequently post hoc multiple comparison with DMRT).
respectively (Table 2). The proximate composition of M. rosenbergii offered diets with various levels of L. sporogenes is given in Table 2. There was a decrease (P < 0.05) in the moisture content while the protein, amino acid, carbohydrate, lipid and ash contents increased (P < 0.05) in all the experimental groups. The highest increase (P < 0.05) in total protein, free amino acid, total carbohydrate, total lipids and ash contents were recorded in the 4% L. sporogenes incorporated diet, followed by 3%, 2% and 1% and the lowest increase was recorded in the control. These differences in proximate composition were statistically significant (P < 0.05), except the ash. It has been reported that in M. rosenbergii PL fed with B. subtilis incorporated diets, there was significant improvement in the proximate composition of body tissues (Seenivasan et al., 2012b). Seenivasan et al., 2011 have reported that with the commercial probiotic, Binifit supplemented diets; there was significant improvement in the proximate composition of body tissues in M. rosenbergii PL. The beneficial effects of probiotics in M. rosenbergii PL culture have also been supported by Venkat et al. (2004), Suralikar and Sahu (2001) and Shinde et al. (2008). Survival performance Under the four levels of L. sporogenes supplement, among the experimental groups, prawns exhibited normal behaviour and no cannibalism was observed. Therefore, L. sporogenes incorporated feed was accepted by the prawns at all levels and showed a significantly higher survival rate. The survival rate was significantly (P < 0.05) higher in M. rosenbergii PL fed with 4% L. sporogenes incorporated diets when compared with other experimental groups (Table 3). Similar improved survival has been reported in M. rosenbergii PL fed with Biogen supplemented diets (Saad et al., 2009) and diets supplemented
Table 3
with different probiotics (Shinde et al., 2008) and in the Indian white shrimp, Fenneropenaus indicus after feeding with Bacillus (Ziaei-Nejad et al., 2006). In general, L. sporogenes has the ability to attach with the gut epithelium and establish there. In the present study, the higher survival rate observed in L. sporogenes incorporated diet fed PL may be due to the administration of significant changes in the proportion of the population of the gut micro flora. This may be exerted by the elimination of harmful bacteria due to establishment of the beneficial probionts, L. sporogenes in the intestine of M. rosenbergii. Or, colonization of L. sporogenes may be dominant over harmful bacteria. By their large presence, saturate the adhesion receptors and prevent the pathogenic bacteria from attachment and colonization (Vine et al., 2004; Seenivasan et al., 2012a). Growth performance L. sporogenes supplement diet appeared to provide beneficial effects on the growth of M. rosenbergii PL. The results indicated that the prawn which had access to diet containing L. sporogenes, exhibited greater growth than that of the control diet (Table 3). There were significant differences in growth increment (P < 0.05) between different experimental and control groups. Final body weight gain, specific growth rate and food conversion ratio in PL offered with L. sporogenes incorporated diets, particularly at 4%, were superior to those of other experimental groups as well as control. Thus, inclusion of L. sporogenes >1% is recommended. Similar results on significant improvement in growth, weight gain and SGR in M. rosenbergii fed with bio-encapsulated L. sporogenes were reported by Seenivasan et al. (2012a). It has been reported that the growth performance, SGR and weight gain were higher in L. sporogenes, B. subtilis and S. cerevisiae supplemented diet
Food indices of M. rosenbergii PL fed with L. sporogenes supplemented diet.
Parameters Survival rate (%) Weight gain (g) Specific growth rate (%) Condition factor (%) Food conversion ratio (g) Food conversion efficiency (%) Protein efficiency rate (g)
Control
1% LS c
77.50 ± 2.50 0.82 ± 0.06c 0.827 ± 0.30c 1.94 ± 0.20a 3.40 ± 0.28a 0.859 ± 0.06b 0.642 ± 0.13b
2% LS c
80.00 ± 2.50 0.85 ± 0.02c 0.841 ± 0.028c 1.73 ± 0.14ab 2.93 ± 0.22ab 0.850 ± 0.08b 0.755 ± 0.11ab
3% LS b
85.00 ± 2.50 0.90 ± 0.04c 0.864 ± 0.029bc 1.63 ± 0.18ab 2.77 ± 0.33ab 0.877 ± 0.10b 0.801 ± 0.09ab
4% LS ab
87.50 ± 2.50 1.01 ± 0.07b 0.911 ± 0.036b 1.45 ± 0.10b 2.55 ± 0.48ab 0.959 ± 0.05b 0.868 ± 0.07a
F-value a
90.00 ± 2.50 1.22 ± 0.09a 0.989 ± 0.039a 1.57 ± 0.22b 2.36 ± 0.80a 1.120 ± 0.12a 0.939 ± 0.10a
12.90 38.65 12.09 3.40 2.15 5.20 3.67
Each value is a mean ± SD of three replicate analyses, within each row means with different superscript letters are statistically significant P < 0.05 (one way ANOVA and subsequently post hoc multiple comparison with DMRT).
Please cite this article in press as: Seenivasan, C. et al., Effect of Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of freshwater prawn Macrobrachium rosenbergii post larvae (2014), http://dx.doi.org/10.1016/ j.jobaz.2013.12.002
L. sporogenes on growth promotion of prawn Table 4
5
Energy budget of M. rosenbergii PL fed with L. sporogenes supplemented diet (values are expressed as k cal/g/day).
Parameters
Control
1% LS
2% LS
3% LS
4% LS
F-value
Feeding rate Absorption rate Conversion rate NH3 Excretory rate Metabolic rate
0.466 ± 0.029c 0.402 ± 0.033c 0.220 ± 0.022b 0.009 ± 0.002b 0.194 ± 0.014b
0.480 ± 0.0320c 0.429 ± 0.030c 0.232 ± 0.027b 0.010 ± 0.003b 0.210 ± 0.017ab
0.511 ± 0.027bc 0.465 ± 0.028bc 0.255 ± 0.031b 0.012 ± 0.002b 0.224 ± 0.019ab
0.549 ± 0.026b 0.508 ± 0.048ab 0.286 ± 0.026ab 0.015 ± 0.003ab 0.237 ± 0.026a
0.605 ± 0.031a 0.568 ± 0.0.41a 0.345 ± 0.034a 0.016 ± 0.001a 0.239 ± 0.021a
11.51 9.59 5.56 4.63 2.75
Each value is a mean ± SD of three replicate analyses, within each row means with different superscript letters are statistically significant P < 0.05 (one way ANOVA and subsequently post hoc multiple comparison with DMRT).
fed M. rosenbergii PL (Seenivasan et al., 2012c). It has been also reported that the growth, SGR and weight gain were increased in bioencapsulated Lactobacillus spp. fed fish, Carassius auratus and Xiphophorus helleri (Ashim et al., 2009). According to Shinde et al. (2008), different probiotics supplemented diets had improved growth rate, weight gain, FCR and FER of M. rosenbergii PL. Therefore, in the present study, improvement of growth and food conversion may be due to L. sporogenes. In addition to growth, the feed conversion efficiency, protein efficiency rate and condition factor were also significantly better (P < 0.05) in PL offered diets with L. sporogenes when compared with control. A possible explanation for the increased performance as a result of the addition of the probiotic is the improvement in digestibility, which may in turn explain the observed better growth rate and the efficiency of feed conversion. Otherwise, probiotics influence digestive processes by enhancing microbial enzyme activity, and consequently improving the digestion, absorption and utilization of the feed (Bomba et al., 2002). Similar observations have been reported by De Schrijver and Ollevier (2000) in turbot, S. maximus fed with the bacteria, Vibrio proteolyticus incorporated diet and by Lara-Flores et al. (2003) in tilapia fry fed with 0.1% probiotics incorporated diets, which improved animal growth and apparent protein digestibility. The higher PER recorded in the present study indicates that the diets with probiotics have significantly improved dietary protein and energy utilization by M. rosenbergii. Venkat et al. (2004) showed a similar improvement in the PER in M. rosenbergii fed probiotics supplemented diets; Bagheri et al. (2008) observed significant improvements of PER in rainbow trout fry (1 g) fed with administration of B. licheniformis + B. subtilis at levels P log 9 cells g1. In the present study, the results indicated that the L. sporogenes incorporation showed positive effect of growth promotion in M. rosenbergii PL. Therefore, the overall growth was higher particularly in 4% L. sporogenes incorporated diet fed M. rosenbergii. Energy utilization performance In the present study, the energy budget of M. rosenbergii offered diets with various levels of L. sporogenes inclusions is given in Table 4. There were significant increases in the feeding rate, absorption rate, conversion rate and NH3 execratory rate in the experimental groups, when compared with the control (P < 0.05). The observed increment was higher at 4% L. sporogenes incorporated diet followed by 3%, 2%, and 1%, and the lowest value was recorded in the control group. It has been reported that the growth rate and feed energy utilization were increased in Saccharomyces cerevisiae supplemented diet fed M. rosenbergii PL (Seenivasan et al., 2013). It has also been
reported that the growth rate performance and nutrient energy utilization were higher in L. sporogenes, B. subtilis and S. cerevisiae inclusion diet fed M. rosenbergii PL (Seenivasan et al., 2012b–e). It has also been reported that the growth rate performance and nutrient energy utilization were higher in Binifit inclusion pellet fed M. rosenbergii PL (Seenivasan et al., 2011). Dhanaraja et al. (2010) reported that probiotics L. acidophilus and yeast S. cerevisiae supplemented diets had improved the energy budget of Koi Carp, C. carpio. It has been reported that the growth rate and feed energy utilization were increased in S. cerevisiae supplemented diet fed Nile tilapia, O. niloticus (Abdel-Tawwab et al., 2008). It has also been reported that the growth rate performance and nutrient energy utilization were higher in Biogen inclusion diet fed Nile tilapia (Haroun et al., 2006). Immanuel et al. (2003) reported that the probiotics Lactobacillus and yeast supplemented diets had improved the energy utilization of pearl spot Etroplus suratensis. From the above discussion, it may be concluded that L. sporogenes incorporation has significantly improved survival, growth, SGR, FCR, PER, FCE, biochemical constituents and energy utilization in post larvae of M. rosenbergii. Therefore, L. sporogenes can be supplemented in formulated diets for the maintenance of healthy M. rosenbergii at nursery and grows out in ponds. Also, M. rosenbergii can be used in farm management at a small scale level, and consequently, inland aquaculture of Macrobrachium may be promoted in a sustainable manner.
Acknowledgement The Bharathiar University, Coimbatore, Tamil Nadu, India is gratefully acknowledged for the financial support provided in the form of University Research Fellowship to Mr. C. Seenivasan. We thank Dr. G. Immanuel, Assistant Professor, Centre for Marine Sciences and Technology, Manonmaniam Sundaranar University, Tamil Nadu, India for providing his subject expertise to conduct the experiment. References Abdel-Tawwab, M., Abdel-Rahman, A.M., Ismael, N.E.M., 2008. Evaluation of commercial live baker’s yeast, Saccharomyces cerevisiae as a growth and immunity promoter for fry Nile tilapia, Oreochromis niloticus (L.) challenged in situ with Aeromonas hydrophila. Aquaculture 280, 185–189. APHA, 2005. Standard Methods for the Examination of Water and Wastewater, 20th ed. American Public Health Association, New York.
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Please cite this article in press as: Seenivasan, C. et al., Effect of Lactobacillus sporogenes on survival, growth, biochemical constituents and energy utilization of freshwater prawn Macrobrachium rosenbergii post larvae (2014), http://dx.doi.org/10.1016/ j.jobaz.2013.12.002