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Research Article - (2024) Volume 15, Issue 1
Evaluation of Acid-Insoluble Ash and Crude Fibre as Internal Markers in In Vivo Digestibility Studies: A Comparison in Three Cyprinid Species
Gangadhar Barlaya1* and Perar Keshavanath22Department of Aquaculture, College of Fisheries, Karnataka Veterinary Animal and Fisheries Sciences University, Mangaluru, India
Received: 09-Nov-2023, Manuscript No. JARD-23-23862; Editor assigned: 13-Nov-2023, Pre QC No. JARD-23-23862 (PQ); Reviewed: 27-Nov-2023, QC No. JARD-23-23862; Revised: 04-Dec-2023, Manuscript No. JARD-23-23862 (R); Published: 11-Dec-2023, DOI: 10.35248/2155-9546.24.15.828
Abstract
Though there are several reports of estimation of apparent nutrient digestibility values using different markers in fish nutritional studies, no study is available wherein the same nutrient digestibility was estimated using two or more markers. The study aimed at comparing the apparent Digestibility Coefficient (ADC) of nutrients, estimated using the markers-Acid-Insoluble Ash (AIA) and Crude Fibre (CF), in order to assess the suitability of digestibility markers. The apparent crude protein and fat digestibility values of experimental diets estimated in three indoor in vivo digestibility studies with cyprinids (Labeo fimbriatus, L. rohita and Hypselobabus pulchellus) using AIA and CF are compared. In these studies, graded dietary levels of cottonseed meal, azolla meal and moringa leaf meal, respectively were evaluated. Apparent digestibility coefficients for protein and fat were calculated from protein and fat in feed and fecal matter, using analyzed AIA and CF contents as digestibility markers. In all the cases, ADC of both crude protein and fat were higher when estimated using AIA as compared to those estimated using CF. However, the general trend in the pattern of variation of ADC values among the different treatments remained almost the same, irrespective of the marker used. The results indicate that AIA as an internal marker is the most appropriate on the basis of the degree of precision that could be achieved in quantification of digestibility. The findings of this study are expected to help fish nutritionists in selection of suitable markers for in vivo digestibility studies.
Keywords
Acid-insoluble ash, Apparent digestibility coefficient, Crude fibre, Digestibility marker, Nutrient digestibility, Carp fish, Food quality, Nutritional food markers
Abbreviations
ADC: Apparent Digestibility Coefficient; AIA: Acid-Insoluble Ash; CF: Crude Fibre; HROM: Hydrolysis Resistant Organic Matter; CSM: Cotton Seed Meal; APD: Apparent Protein Digestibility; AFD: Apparent Fat Digestibility
Introduction
Determination of digestibility of different feed ingredients is significant for developing cost-effective diet formulas, assessing the quality of the ingredients, and preventing the release of nutrients into the environment that could harm the ecosystem [1,2]. Digestibility can be estimated both in vitro as well as in vivo. The former involves incubation of the test ingredient with the crude digestive enzyme extracted from the gut of the test fish [3]. Compared to in vivo procedures, in vitro techniques may be able to forecast changes in digestibility resulting from modifications in food processing more quickly and affordably. However, in vivo data on digestibility are needed to validate the data obtained through in vitro studies [4]. When using in vitro techniques, the responses are more extreme than when fish naturally digest food, and they release nutrients that wouldn’t otherwise be available [5]. When comparing in vitro methods to in vivo approaches, Marletta L, et al. found that there is a propensity to overestimate the digestibility of leguminous plants [6]. Though in vivo estimation of digestibility is expensive and time-consuming, it is the most popular method to determine the bioavailability of nutrients from a feed stuff for a given species.
Different types of markers have been used in in vivo digestibility studies with fish. The external (exogenous) markers include chromic oxide, rare earth metal oxides such as ytterbium oxide and yttrium oxide, and hydrocarbon markers such as cholestane [7-9]. Acid-insoluble ash (AIA)/hydrolysis resistant ash, cellulose, Hydrolysis Resistant Organic Matter (HROM) and Crude Fibre (CF) have been used as internal (endogenous/natural/indigenous) markers for estimating nutrient digestibility [10-15]. There is ongoing debate about whether internal or exterior indicators are more suited and/or reliable and which specific indicators should be used. The majority of the research suggests that indigenous markers-which are present in significant amounts in the diet-are preferable to exogenous markers [16].
The use of internal markers such as AIA and CF has been recommended for the estimation of apparent digestibility coefficient (ADC) [17]. AIA has been reported to be an effective internal marker in digestibility studies with rainbow trout, Oncorhynchus mykiss; Arctic charr, Salvelinus alpinus; channel catfish, Ictalurus punctatus; and tilapia, Oreochromis aureus [18-21]. However, results obtained are not always consistent. When used with rainbow trout, AIA has been shown to provide digestibility coefficients that are comparable to those produced using chromic oxide, greater than those obtained using chromic oxide, or even lower [7,18,22]. Some researchers have opined limited use of AIA through their studies with different fish species [7,14,22].
CF has been found to be a more reliable marker than AIA for the cichlid Etroplus suratensis [11]. Several studies have shown CF as an effective marker [7,23].
Though AIA and CF have been used by several researchers as internal markers for digestibility estimations, no literature comparing the results obtained using these markers is available. The study aimed at comparing the ADC of nutrients, estimated using the marker-acid-insoluble ash and crude fibre, in order to assess the suitability of markers for in vivo digestibility studies. The present study compares the ADCs obtained for dietary protein and fat using AIA and CF as markers in three digestibility studies conducted with Labeo fimbriatus, L. rohita and Hypselobabus pulchellus, to find out the more appropriate one among them on the basis of the degree of precision that could be achieved in quantification. An effort has also been made to compare the ADC values obtained by several authors using these 2 markers.
Materials and Methods
Applicable institutional guidelines for the care and use of animals were followed by the authors. In the present study, the digestibility values from three studies were taken for comparison. The digestibility values obtained using AIA as the marker were taken from two published papers and a communicated paper [24,25]. A brief explanation of preparation of experimental diets and conducting the digestibility trials is given for better understanding.
Preparation of experimental diets
The basal diet (control) for experiment 1 with L. fimbriatus was prepared with rice bran (45%) and groundnut cake (45%) (Table 1). The binder used was finger millet (9%). The diets had Cotton Seed Meal (CSM) at 10%, 20%, 30% and 40% levels replacing groundnut oilcake and rice bran in the basal diet [26]. For preparing the diets, the following steps were followed. All the ingredients other than rice bran were dried and powdered. After sieving the ingredients (0.5 mm), required quantity of the ingredients was mixed with hot water. A dough was made and passed through a pelletiser with 2 mm die. The pellets thus obtained were sun dried. The diets for the experiments 2 (L. rohita) and 3 (H. pulchellus) were also prepared in the same manner excepting that dried Azolla powder or Moringa leaf meal was used in place of CSM. The percentage incorporation of other ingredients in the 3rd experiment is not given since the whole experimental data is communicated to a journal.
| Ingredients | Control | 10% CSM/Azolla | 20% CSM/Azolla | 30% CSM/Azolla | 40% CSM/Azolla |
|---|---|---|---|---|---|
| Groundnut cake | 45 | 40 | 35 | 30 | 25 |
| Rice bran | 45 | 40 | 35 | 30 | 25 |
| Finger millet | 8 | 8 | 8 | 8 | 8 |
| CSM | 0 | 10 | 20 | 30 | 40 |
| Vitamin and mineral mixture | 2 | 2 | 2 | 2 | 2 |
Note: CSM: Cotton Seed Meal.
Table 1: Ingredient proportion (%) of Cotton Seed Meal (CSM) incorporated diets for evaluation in L. fimbriatus.
Experimental design
Digestibility studies were conducted in 50 l, plastic tanks provided with aeration [24]. In the 1st study, 15 tanks were stocked with 10 advanced fingerlings of L. fimbriatus (fimbriatus) having body weight ranging from 7.39 g-10.88 g. After the initial acclimation for 10 days with control diet, fish were fed the 5 diets at 5% of body weight, every morning at 10:00 h, in triplicate tanks. The remaining pellets were siphoned out after 3 hours. The faecal matter from each tank was collected on the following day morning, by filtering the tank water with a nylon cloth (15 µm), dried and stored for proximate analysis. After faecal matter removal, nearly 50% of water from each tank was replaced with freshwater. This process was continued for a period of 45 days to obtain adequate quantity of dried faecal matter. Similar protocol was followed in the other 2 experiments. The fish used in the 2nd and 3rd experiment were L. rohita (rohu) and H. pulchellus (pulchellus) respectively, with stocking weight ranging from 8.50 g to 11.00 g and 3.25 g to 3.75 g, respectively [25].
Crude protein and fat content of the pelleted feed and faecal matter were analyzed [27]. Both AIA and CF were used as the reference markers [21,23,28,29]. AIA in diet and fecal matter was determined gravimetrically after drying, ashing, boiling of ash in hydrochloric acid (1:2), filtering and washing of the hot hydrolysate and re-ashing [18]. CF was estimated gravimetrically after chemical digestion and solubilization of other materials present. The fiber residue weight was then corrected for ash content after ignition [27].
Maynard et al, formula was used to calculate apparent nutrient digestibility as per the following equations [30].
Total dry matter digestibility (%)=100–[100 × % marker in feed/% marker in feces]
Nutrient digestibility (%)=100–[100 × %marker in feed/% marker in feces] × [%Nutrient in feces/% Nutrient in feed]
Statistical analyses
Analysis of variance was used for testing the data for statistical difference and Duncan’s multiple range test (P=0.05) was applied to rank the treatment means tested for significance [31].
Results
In all the digestibility trials, fish readily accepted the prepared diets and no mortality was observed. Table 2 gives the ADCs of crude protein and fat of CSM incorporated diets fed to fimbriatus.
| Acid-Insoluble Ash (AIA) as the marker | Crude Fibre (CF) as the marker | |||
|---|---|---|---|---|
| Diets | Protein | Fat | Protein | Fat |
| Control | 83.01 ± 2.03a | 91.60 ± 0.26ab | 68.93 ± 0.79a | 86.92 ± 0.23d |
| 10% CSM | 85.37 ± 1.49ab | 93.15 ± 0.45c | 72.38 ± 1.95ab | 87.06 ± 0.44d |
| 20% CSM | 86.03 ± 0.78b | 92.79 ± 0.79c | 71.57 ± 0.70bc | 82.96 ± 2.83bc |
| 30% CSM | 86.69 ± 0.28b | 92.42 ± 1.40bc | 73.75 ± 2.19c | 83.35 ± 1.32dc |
| 40% CSM | 86.15 ± 1.73b | 89.68 ± 0.80a | 69.58 ± 1.13bc | 77.28 ± 0.24a |
Note: CSM: Cotton Seed Meal, a,b,c: Figures in the same column with different superscripts differ significantly (P<0.05).
Table 2: Apparent digestibility coefficient (%, Mean ± SD, n = 3) of crude protein and fat calculated using Acid-Insoluble Ash (AIA) and Crude Fibre (CF) in Cotton Seed Meal (CSM) incorporated diets fed to L. fimbriatus.
The Apparent Protein Digestibility (APD) estimated using AIA varied between 83.01% and 86.69%, while that estimated using CF varied from 68.93% to 73.75%. The apparent fat digestibility (AFD) estimated using AIA ranged between 89.68% and 93.15%, while that estimated using CF fluctuated from 77.28% to 87.06%. It is evident that values were lower when CF was used as the marker compared to those obtained using AIA as the marker. However, the general response pattern in ADCs remained almost the same with both the markers. In the 2nd study with Azolla, the APD estimated in rohu using AIA varied between 73.45% and 87.97%, while that estimated using CF ranged between 67.28% and 82.31%. The AFD estimated using AIA fluctuated from 93.60% to 99.48%, while that estimated using CF varied between 92.13% and 99.28% (Table 3).
| AIA as the marker | CF as the marker | |||
|---|---|---|---|---|
| Diets | Protein | Fat | Protein | Fat |
| Control | 87.97 ± 0.40c | 99.39 ± 0.12c | 82.31 ± 0.52c | 99.11 ± 0.19cd |
| 10% Azolla | 85.72 ± 0.47c | 99.48 ± 0.03c | 80.10 ± 2.06c | 99.28 ± 0.06d |
| 20% Azolla | 85.17 ± 0.74c | 96.76 ± 0.79c | 80.13 ± 1.68c | 95.71 ± 2.78bc |
| 30% Azolla | 77.59 ± 1.43b | 93.60 ± 1.02b | 74.14 ± 1.50b | 92.63 ± 1.08ab |
| 40% Azolla | 73.45 ± 1.68a | 93.61 ± 0.65a | 67.28 ± 2.96a | 92.13 ± 0.77a |
Note: CF: Crude Fibre; AIA: Acid-Insoluble Ash,a,b,c,d: Figures in the same column with different superscripts differ significantly (P<0.05).
Table 3: Apparent digestibility coefficient (%, Mean ± SD, n = 3) of crude protein and fat calculated using Acid-Insoluble Ash (AIA) and crude fibre (CF) in azolla incorporated diets fed to L. rohita.
In this study also, the APD values were lower when CF was used as the marker compared to those obtained using AIA as the marker. However, the AFD values obtained with CF were almost equal to those obtained with AIA. In the 3rd experiment, the APD and AFD obtained with AIA in pulchellus varied between 87.23% and 94.75% and 91.55% and 98.16%, while the APD and AFD values calculated using CF ranged from 64.49% to 68.78% and from 86.91% to 91.01%, respectively (Table 4). Lower digestibility values with CF compared to AIA are apparent here also.
| AIA as the marker | CF as the marker | |||
|---|---|---|---|---|
| Diets | Protein | Fat | Protein | Fat |
| Control | 92.34 ± 2.18a | 97.83 ± 1.75a | 65.43 ± 1.67ab | 89.95 ± 0.30a |
| 5% MLM | 87.23 ± 1.74a | 91.55 ± 1.39a | 68.78 ± 1.24c | 91.01 ± 0.33a |
| 10% MLM | 88.59 ± 1.71a | 92.50 ± 1.94a | 67.00 ± 1.16bc | 90.12 ± 2.12a |
| 15% MLM | 94.75 ± 2.82b | 98.16 ± 0.93a | 67.57 ± 0.41bc | 89.72 ± 1.56a |
| 20% MLM | 93.09 ± 1.69a | 97.73 ± 1.71a | 64.49 ± 0.89a | 89.47 ± 1.59a |
| 25% MLM | 92.35 ± 1.01a | 97.08 ± 1.85a | 65.13 ± 1.81a | 86.91 ± 0.66a |
| 30% MLM | 92.91 ± 2.67a | 97.49 ± 1.84a | 64.98 ± 0.41a | 87.51 ± 0.40a |
Note: MLM: Moringa Leaf Meal; a,b,c: Apparent digestibility coefficients; AIA: Acid-Insoluble Ash; CF: Crude Fibre.
Table 4: Apparent digestibility coefficient (%, Mean ± Standard Deviation (SD), n = 3) of crude protein and fat calculated using Acid-Insoluble Ash (AIA) and Crude Fibre (CF) in Moringa Leaf Meal (MLM) incorporated diets fed to H. pulchellus.
Discussion
From the results of this study it is clear that ADC values obtained with AIA are higher compared to those obtained with CF which is evident from other studies as well. Tables 5 gives the ADC of protein and fat reported in earlier in vivo digestibility studies using AIA and CF as markers. Table 6 summarizes the APD and AFD values listed in Table 5. Among the 7 studies which used AIA as marker, 71% reported APD in the range of 90%-100%, whereas the rest 29% recorded APD of 80%-90%. Among the listed studies that used CF as marker, none reported APD ranging from 90%-100%. Moreover, a small percentage even reported APD in the lowest range of 50%-60%. With regard to AFD, studies with AIA recorded values ranging from 80-100%, while those with CF reported values varying from 70%-100%.
| Marker | Apparent Digestibility Coefficient (ADC) of protein (%, range) | Apparent Digestibility Coefficient (ADC) of fat (%, range) | Major ingredients | Species | Treatment | Reference |
|---|---|---|---|---|---|---|
| Acid-Insoluble Ash (AIA) | 68.61-92.62 | 74.00-86.65 | Fishmeal, blood meal, soybean meal, corn | Clarias gaiepinus | Dietary incorporation of brewer’s spent grain replacing corn at 0 to 100% | [41] |
| Acid-Insoluble Ash (AIA) | 82.96-89.98 | 87.97-91.13 | Fishmeal, soybean meal, corn, cassava starch | Oreochromis niloticus | Dietary incorporation of Jackbean meal replacing SBM at 10 and 40% | [42] |
| Acid-Insoluble Ash (AIA) | 78.04-92.22 | 74.76-91.24 | Fishmeal, soybean meal, corn, starch | Oreochromis niloticus | Dietary incorporation of Jatropha seed meal replacing SBM at 20% and 40% | [43] |
| Acid-Insoluble Ash (AIA) | 87.58-91.36 | 84.04-89.05 | Fishmeal, soybean meal, maize, cassava starch | Clarias gaiepinus | Dietary incorporation of cooked sunflower seed meal replacing SBM at 15%, 30% and 45% | [44] |
| Acid-Insoluble Ash (AIA) | 86.80-92.09 | 84.97-88.54 | Fishmeal, soybean meal, corn meal, starch | Clarias gaiepinus | Dietary incorporation of toasted sunflower seed meal replacing SBM at 15%, 30% and 45% | [45] |
| Acid-Insoluble Ash (AIA) | 82.7-92.5 | 82.7-92.5 | Fishmeal, soybean meal, Corn, wheat middlings | Oreochromis mykiss | Dietary incorporation of sheep skin and alpaca skins hydrolysate at 30% | [46] |
| Acid-Insoluble Ash (AIA) | 81.02-83.00 | 85.62-87.50 | Wheat bran, rice bran, wheat middling, maize, sunflower meal, maize gluten meal, poultry meal, soybean meal | Oreochromis niloticus | Feeding balanced lysine diet and low lysine diet | [47] |
| Crude fibre | 81.73-88.94 | 77.12-88.82 | Fishmeal, groundnut cake, rice bran, tapioca | Cyprinus carpio | Dietary incorporation of Spirulina powder replacing FM at 25%-100% | [48] |
| Crude fibre | 73.18-86.59 | 48.00-76.14 | Fishmeal, groundnut cake, rice bran, tapioca | Cyprinus carpio | Dietary incorporation of 19 Norethindrone at 0.25 ppm-1.00 ppm | [49] |
| Crude fibre | 50.14-58.32 | 54.07-70.47 | Fishmeal, groundnut cake, rice bran, tapioca | Cyprinus carpio | Dietary incorporation of NaCl at 0.5%-2.0% | [50] |
| Crude fibre | 66.09-78.39 | 50.29-74.98 | Fishmeal, groundnut cake, rice bran, tapioca | Cirrhinus mrigala | Dietary incorporation of NaCl at 0.5%-2.0% | [50] |
| Crude fibre | 74.22-83.29 | 66.70-79.34 | Fishmeal, groundnut cake, rice bran, tapioca | Labeo rohita | Dietary incorporation of Spirulina powder replacing FM at 25%-100% | [51] |
| Crude fibre | 82.76-88.28 | 78.39-83.66 | Fishmeal, groundnut cake, rice bran, tapioca | Catla catla | Dietary incorporation of Spirulina powder replacing FM at 25%-100% | [51] |
| Crude fibre | 59.57-74.84 | 59.97-90.63 | Groundnut cake, rice bran, finger millet | Labeo calbasu | Dietary incorporation of azolla powder replacing basal diet at 10%-40% | [52] |
| Crude fibre | 59.17-74.55 | 87.25-90.15 | Groundnut cake, rice bran, finger millet | Labeo fimbriatus | Dietary incorporation of azolla powder replacing basal diet at 10%-40% | [53] |
| Crude fibre | 47.63-69.56 | 77.07-81.42 | Groundnut cake, rice bran, finger millet | Cyprinus carpio | Dietary incorporation of azolla powder replacing basal diet at 10%-40% | [53] |
| Crude fibre | 72.27-83.35 | 84.31-94.73 | Groundnut cake, rice bran, finger millet | Labeo fimbriatus | Dietary incorporation of soybean powder replacing basal diet at 10%-40% | [53] |
| Crude fibre | 71.84-83.00 | 79.31-85.54 | Groundnut cake, rice bran, finger millet | Cyprinus carpio | Dietary incorporation of soybean powder replacing basal diet at 10%-40% | [53] |
| Crude fibre | 72.49-78.67 | 75.78-83.71 | Groundnut cake, rice bran, finger millet | Labeo fimbriatus | Dietary incorporation of silkworm pupa powder replacing basal diet at 10-40% | [53] |
| Crude fibre | 69.71-77.64 | 85.52-95.28 | Groundnut cake, rice bran, finger millet | Cyprinus carpio | Dietary incorporation of silkworm pupa powder replacing basal diet at 10%-40% | [53] |
| Crude fibre | 44.91-75.45 | 82.79-96.83 | Groundnut cake, rice bran, finger millet | Catla catla | Dietary incorporation of azolla powder replacing basal diet at 10%-40% | [54] |
| Crude fibre | 68.76-84.96 | 80.93-88.32 | Groundnut cake, rice bran, finger millet | Catla catla | Dietary incorporation of soybean powder replacing basal diet at 10%-40% | [54] |
| Crude fibre | 67.17-70.87 | 84.18-94.66 | Groundnut cake, rice bran, finger millet | Catla catla | Dietary incorporation of silkworm pupa powder replacing basal diet at 10%-40% | [54] |
Table 5: Apparent Digestibility Coefficient (ADC) of protein and fat reported in earlier in vivo digestibility studies using Acid-Insoluble Ash (AIA) and Crude Fibre (CF) as markers.
| 90%-100% | 80%-90% | 70%-80% | 60%-70% | 50%-60% | |
|---|---|---|---|---|---|
| ADC of protein estimated with CF | 0 | 43.75% | 43.75% | 6.25% | 6.25% |
| ADC of fat estimated with CF | 37.5% | 37.5% | 25% | 0 | 0 |
| ADC of protein estimated with AIA | 71% | 29% | 0 | 0 | 0 |
| ADC of fat estimated with AIA | 43% | 57% | 0 | 0 | 0 |
Note: ADC: Apparent Digestibility Coefficient; CF: Crude Fibre; AIA: Acid-Insoluble Ash.
Table 6: Range (%) of protein and fat Apparent Digestibility Coefficient (ADC) values obtained with Crude fibre (CF) and Acid-Insoluble Ash (AIA) as listed in Table 5.
CF has been shown to be assimilated to a very small extent at least in certain fish species [32,33]. Although fish are unable to synthesize cellulase internally, their digestive tracts include microbial communities that aid in the digestion of plant materials [34,35]. The lower ADC values obtained in the present study with CF as the marker are attributable to the exogenous cellulase in fish gut.
Another important observation is the higher difference between the ADC values obtained through AIA and CF in the 3rd study with pulchellus compared to those obtained with the other two species. In nature, pulchellus is mainly a herbivorous, marginal submerged vegetative feeder subsisting on Chara, Hydrilla, Vallisneria and Ceratophyllum sp. In the gut analysis study by David A et al, decayed tissues of higher aquatic plants as well as grass blades were also detected [36]. Basavaraja N et al, observed that the captive stock of pulchellus very well accepts Napier grass, apart from artificial feed [37]. Hence, the species is expected to utilize plant material efficiently. Our study on the same species with dietary incorporation of aquatic plants-Azolla sp. and Vallisneria sp. has corroborated this hypothesis [38]. Being able to consume aquatic vegetation, pulchellus may be able to break down some quantity of fibre in the gut, leading to lower ADC values estimated through CF.
It is very unlikely that the AIA fraction can pass through the gut wall and hence, the higher ADC values. The advantages of using AIA as a marker includes the low cost and ease of measurement using basic laboratory equipment [10]. When utilizing AIA as a marker, it was suggested that it is preferable for the diet to contain AIA in excess of 0.75% [39]. In the present study, the AIA values ranged from 0.96% to 1.27 in the 1st study with CSM, 1.78% to 4.68% in the 2nd one with Azolla and 0.70% to 0.82% in the 3rd experiment with Moringa leaf meal. Sales J et al and Atkinson et al, opined that AIA is a trustworthy marker and could be used to accurately assess the digestibility of nutrients [18,40]. However, when pure ingredients are used for making experimental diets, AIA being primarily silica may not be present in sufficient quantity for accurate analysis. In such cases CF can be an alternative [41-54].
Conclusion
Though there are several reports of estimation of apparent nutrient digestibility values using different markers in fish nutritional studies, no study is available wherein the same nutrient digestibility was estimated using two or more markers. In the present study, the apparent crude protein and fat digestibility values of experimental diets estimated in three indoor in vivo digestibility studies with cyprinids (Labeo fimbriatus, L. rohita and Hypselobabus pulchellus) using AIA and CF are compared. The results of the present study indicate that usage of AIA as an internal marker is the most appropriate on the basis of the degree of precision that could be achieved in quantification of digestibility in carps. The findings of this study are expected to help fish nutritionists in selection of suitable markers for in vivo digestibility studies.
Acknowledgment
The authors are grateful to the Director, ICAR-Central Institute of Freshwater Aquaculture, Bhubaneswar for the facilities.
Conflicts of Interest
There are no conflicts of interest.
References
- Gatlin D, Hardy R. Manipulations of diets and feeding to reduce losses of nutrients in intensive aquaculture. Aquaculture and the Environment in the United States. 2002:155-165.
- Montano-Vargas J, Shimada A, Vásquez C, Viana MT. Methods of measuring feed digestibility in the green abalone (Haliotis fulgens). Aquaculture. 2002;213(1-4):339-346.
- Lemos D, Del Toro AN, Córdova-Murueta JH, Garcia-Carreño F. Testing feeds and feed ingredients for juvenile pink shrimp Farfantepenaeus paulensis: In vitro determination of protein digestibility and proteinase inhibition. Aquaculture. 2004;239(1-4):307-321.
- Ezquerra JM, García-Carreño FL, Civera R, Haard NF. pH-stat method to predict protein digestibility in white shrimp (Penaeus vannamei). Aquaculture. 1997;157(3-4):251-262.
- Anderson JS, Lall SP, Anderson DM, McNiven MA. Evaluation of protein quality in fish meals by chemical and biological assays. Aquaculture. 1993;115(3-4):305-325.
- Marletta L, Carbonaro M, Carnovale E. Invitro protein and sulphur amino acid availability as a measure of bean protein quality. J Sci Food Agric. 1992;59(4):497-504.
- Morales AE, Cardenete G, Sanz A, De la Higuera M. Re-evaluation of crude fibre and acid-insoluble ash as inert markers, alternative to chromic oxide, in digestibility studies with rainbow trout (Oncorhynchus mykiss). Aquaculture. 1999;179(1-4):71-79.
- Austreng E, Storebakken T, Thomassen MS, Refstie S, Thomassen Y. Evaluation of selected trivalent metal oxides as inert markers used to estimate apparent digestibility in salmonids. Aquaculture. 2000;188(1-2):65-78.
- Carter CG, Lewis TE, Nichols PD. Comparison of cholestane and yttrium oxide as digestibility markers for lipid components in Atlantic salmon (Salmo salar L.) diets. Aquaculture. 2003;225(1-4):341-351.
- Bowen SH. Digestion and assimilation of periphytic detrital aggregate by Tilapia mossambica. Trans Am Fish Soc. 1981;110(2):239-245.
- De Silva SS, Perera MK. Digestibility of an aquatic macrophyte by the cichlid Etroplus suratensis (Bloch) with observations on the relative merits of three indigenous components as markers and daily changes in protein digestibility. J Fish Biol. 1983;23(6):675-684.
- Buddington RK. Digestion of an aquatic macrophyte by Tilapia zillii (Gervais). J Fish Biol. 1979;15(4):449-555.
- Buddington RK. Hydrolysis-resistant organic matter as a reference for measurement of fish digestive efficiency. Trans Am Fish Soc. 1980;109(6):653-656.
- Tacon AG, Rodrigues AM. Comparison of chromic oxide, crude fibre, polyethylene and acid-insoluble ash as dietary markers for the estimation of apparent digestibility coefficients in rainbow trout. Aquaculture. 1984;43(4):391-399.
- Bob-Manuel FG, Alfred-Ockiya JF. Evaluation of yeast single cell protein (SCP) diets on growth performance, feed conversion and carcass composition of Tilapia Oreochromis niloticus (L.) fingerlings. Afr J Biotechnol. 2011;10(46):9473-7478.
- De Silva SS. Digestibility evaluations of natural and artificial diets. Spec Publ Asian Fish Soc. 1989;4:36-45.
- De Silva SS, Anderson TA. Fish nutrition in aquaculture. Springer Science & Business Media; 1994.
- Atkinson JL, Hilton JW, Slinger SJ. Evaluation of acid-insoluble ash as an indicator of feed digestibility in rainbow trout (Salmo gairdneri). Can J Fish Aquat Sci. 1984;41(9):1384-1386.
- Gurure RM, Moccia RD, Atkinson JL. Apparent digestibility and amino acid availability of six feed ingredients by Arctic charr (Salvelinus alpinus). Bulletin-Aquaculture Association of Canada. 1996:32-34.
- TM S. Digestibility determination in Nile catfish fingerlings using internal and external markers. 1933.
- Goddard JS, McLean E. Acid-insoluble ash as an inert reference material for digestibility studies in tilapia, Oreochromis aureus. Aquaculture. 2001;194(1-2):93-98.
- Perera WM, Carter CG, Houlihan DF. Apparent absorption efficiencies of amino acids in rainbow trout, Oncorhynchus mykiss (Walbaum), fed diets containing bacterial single‐cell protein. Aquac. Nutr. 1995;1(2):95-103.
[Crossref] [Google Scholar] [PubMed]
- Krontveit RI, Bendiksen EÅ, Aunsmo A. Field monitoring of feed digestibility in Atlantic salmon farming using crude fiber as an inert marker. Aquaculture. 2014;426:249-255.
- Barlaya G, Bs Ak, Hegde G, Kannur H. Nutrient digestibility and digestive enzyme activity in fringe lipped carp, Labeo fimbriatus (Bloch, 1795), Fed Diets Containing Cottonseed Meal. Asian Fish Sci. 2021;34(3).
- Gangadhar B, Umalatha H, Hemaprasanth K. Apparent digestibility of dried Azolla powder incorporated diets in rohu Labeo rohita (Hamilton, 1822). Indian Journal of Fisheries. 2021;68(4):153-156.
- Umesh NR, Dathatri K, Nandeesha MC, Gangadhara B, Varghese TJ. Digestibility of dry matter and protein from Spirulina platensis by common carp, Cyprinus carpio, with a note on time of faeces collection in digestibility estimations. In Fish nutrition Research Asia, Proceedings of the Fifth Asian Fish Nutrition Workshop, Asian Fisheries Society, Manilla 1994.81-84.
- Horwitz W. Official methods of analysis. Washington, DC: Association of Official Analytical Chemists; 1975.
- Bob-Manuel FG. Methods used in digestibility evaluation of fish diets: A review and challenges. Continental Journal of Fisheries and Aquatic Science. 2013;7(2):25.
- De Silva SS, Shim KF, Ong AK. An evaluation of the method used in digestibility estimations of a dietary ingredient and comparisons on external and internal markers, and time of faeces collection in digestibility studies in the fish Oreochromis aureus (Steindachner). Reprod Nutr Dev. 1990;30(2):215-226.
- Maynard, L. A., Loosli, J. K. Animal nutrition, 6th edn. McGraw-Hill, New York, USA. 1972
- Duncan DB. Multiple range and multiple F tests. Biometrics. 1955;11(1):1-42.
- Van Dyke JM, Sutton DL. Digestion of duckweed (Lemna spp.) by the grass carp (Ctenopharyngodon idella). J Fish Biol. 1977;11(3):273-8.
- Niederholzer R, Hofer R. The adaptation of digestive enzymes to temperature, season and diet in roach Rutilus rutilus L. and rudd Scardinius erythrophthalmus L. Cellulase. J Fish Biol. 1979;15(4):411-416.
- Saha S, Roy RN, Sen SK, Ray AK. Characterization of cellulase‐producing bacteria from the digestive tract of tilapia, Oreochromis mossambica (Peters) and grass carp, Ctenopharyngodon idella (Valenciennes). Aquaculture Research. 2006;37(4):380-388.
- Li H, Zheng Z, Cong-xin X, Bo H, Chao-yuan W, Gang H. Isolation of cellulose—producing microbes from the intestine of grass carp (Ctenopharyngodon idellus). Environ Biol Fishes. 2010:131-135.
- David A, Rahman MF. Studies on some aspects of feeding and breeding of Puntius pulchellus (Day) and its utility in culturable waters. Journal of the Inland Fisheries Society of India. 1976.
- Basavaraja N. Comments on Hypselobarbus pulchellus part of the articles by Knight et al. (2013a, b) published in JoTT. J Threat Taxa. 2014;6(1):5417-5418.
- Barlaya G, Basumatary P, Raghavendra CH, Kumar BS, Kannur H. Utilization of Azolla and Vallisneria as ingredients in the diet for endangered peninsular carp Hypselobarbus pulchellus (Day, 1870). J Appl Ichthyol. 2022;38(4):448-454.
- Thonney ML, Palhof BA, DeCarlo MR, Ross DA, Firth NL, Quaas RL, et al. Sources of variation of dry matter digestibility measured by the acid insoluble ash marker. J Dairy Sci. 1985;68(3):661-668.
- Sales J, Janssens GP. Acid-insoluble ash as a marker in digestibility studies: A review. J Anim Feed Sci. 2003;12(3):383-401.
- Ugbor ON, Ochang SN, Ikwuemesi JC, Alum-Udensi O, Agbakwuo CA. Apparent nutrient digestibility coefficient of brewer’s spent grain meal based diets in Clarias gariepinus juveniles. Int J Fish Aquac.2018;4(1):28-36
- Jimoh WA, Fagbenro OA, Adeparusi EO. Digestibility coefficients of processed jackbean meal Cannavalia ensiformis (L.) DC for Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) diets. Int j fish aquat 2010;2(4):102-107.
- Jimoh WA, Ayeloja AA, Abubakar MI, Yusuf YO, Shittu MO, Abdulsalami SA. Toasted Jatropha curcas seed meal in Nile tilapia (Oreochromis niloticus) diet: Effect on growth, economic performance, haematology, serum biochemistry and liver histology. Int J Aquat Biol. 2020;8(2):98-108.
- Jimoh WA. Growth, nutrient utilization, body composition, hematology and histopathology of the liver of Clarias gariepinus fed cooked sunflower based diets. Ege Journal of Fisheries and Aquatic Sciences. 2020;37(4):343-351.
- Jimoh WA. Dietary replacement of soybean meal by toasted sunflower seedmeal in the diet of Clarias gariepinus: Effect on growth, body composition, digestibility, haematology and histopathology of the liver. Iraqi J Agric Sci. 2020;51(4):1088-1103.
- Araníbar-Araníbar MJ, Roque-Huanca B, Portocarrero-Prado HS, Rodríguez-Huanca FH, Del Pilar Blanco-Espezúa M, Araníbar-Béjar HK, et al. Nutritive value and digestibility of macronutrients from sheep and alpaca skin hydrolysates as a new alternative in juvenile rainbow trout (Oncorhynchus mykiss) feeding. Fish Aquatic Sci. 2020;23(1):1-7.
- Khalil HS, Momoh T, Al-Kenawy D, Yossa R, Badreldin AM, Roem A, et al. Nitrogen retention, nutrient digestibility and growth efficiency of Nile tilapia (Oreochromis niloticus) fed dietary lysine and reared in fertilized ponds. Aquac Nutr. 2021;27(6): 2320-2332.
- Nandeesha MC, Gangadhar B, Varghese TJ, Keshavanath P. Effect of feeding Spirulina platensis on the growth, proximate composition and organoleptic quality of common carp, Cyprinus carpio L. Aquac Res. 1998;29(5):305-312.
- Gangadhar B, Nandeesha MC, Varghese TJ, Keshavanath P. Effect of 19-norethisterone on growth, biochemical composition and gut digestive enzyme activity of common carp, Cyprinus carpio L. J. Aquacult Trop. 2000;15(1):1-10.
- Nandeesha MC, Gangadhar B, Keshavanath P, Varghese TJ. Effect of dietary sodium chloride supplementation on growth, biochemical composition and digestive enzyme activity of young Cyprinus carpio (Linn.) and Cirrhinus mrigala (Ham.). J Aquacult Trop. 2000;15(2):135-144.
- Nandeesha MC, Gangadhara B, Manissery JK, Venkataraman LV. Growth performance of two Indian major carps, catla (Catla catla) and rohu (Labeo rohita) fed diets containing different levels of Spirulina platensis. Bioresour Technol. 2001;80(2):117-120.
- Gangadhar B, Umalatha H, Hegde G, Sridhar N. Digestibility of dry matter and nutrients from Azolla pinnata by Labeo calbasu (Hamilton, 1822) with a note on digestive enzyme activity. Fishery Technology. 2017;54(2):94-99.
- Gangadhar B, Umalatha H, Ganesh H, Saurabh S, Sridhar N. Digestibility of dry matter and nutrients from three ingredients by the carps, Labeo fimbriatus (Bloch, 1795) and Cyprinus carpio Linnaeus, 1758 with a note on digestive enzyme activity. Indian J Fish. 2017;64(3):75-84.
- Umalatha H, Gangadhar B, Hegde G, Sridhar N. Digestibility of three feed ingredients by Catla catla (Hamilton, 1822). Oceanogr Fish Open Access J. 2018;5(5):99-104.
Citation: Barlaya G, Keshavanath P (2023) Evaluation of Acid-Insoluble Ash and Crude Fibre as Internal Markers in In Vivo Digestibility Studies: A Comparison in Three Cyprinid Species. J Aquac Res Dev. 15:829.
Copyright: © 2023 Barlaya G, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.