Nutritional, physiological and immunological evaluation of cricket (Gryllus bimaculatus) and mealworm (Tenebrio molitor) meals as fish meal substitute in diets for olive flounder (Paralichthys olivaceus) and black porgy (Acanthopagrus schlegelii) juveniles
- 발행기관 강릉원주대학교 일반대학원
- 발행년도 2021
- 학위수여년월 2021. 8
- 학위명 박사
- 학과 및 전공 도움말 일반대학원 해양생물공학과
- 실제URI http://www.dcollection.net/handler/kangnung/000000010993
- UCI I804:42001-000000010993
- 본문언어 영어
초록/요약 도움말
국문요약 1. 넙치 치어 사료내 어분대체원으로서 쌍별귀뚜라미의 이용성 본 연구는 어분 대체원으로 쌍별귀뚜라미를 첨가한 배합 사료가 넙치 치어의 성장, 체조성 및 면역에 미치는 영향을 조사하기 위해 수행되었다. 실험사료의 주 단백질원으로 정어리어분 및 멸치어분을 사용하였으며, 지질원으로 어유 및 대두유를 사용하였다. 그리고 사료 내 쌍별귀뚜라미 첨가에 따른 넙치의 반응을 알아보기 위해 대조사료(Con)의 조성물 중 어분, 소맥분 및 대두유의 첨가비를 감소시키면서 쌍별귀뚜라미를 각각 13, 26, 39 및 52% 첨가하여 총 5종류의 실험사료를 설계하였다. 사육실험은 평균 33.5±0.01 g 전후의 넙치를 선별하여 150L 원형수조에 각각 20마리씩 3반복으로 수용하여 8주간 수행하였다. 쌍별귀뚜라미의 첨가 정도를 달리한 배합사료를 8주간 공급하여 사육한 결과 넙치의 생존율은 유의적인 차이가 없었다(P > 0.05). 그러나 넙치의 증체율 및 일간성장율은 CR20 실험구가 CR80 실험구보다 유의적으로 높았다(P < 0.05). 외관상 건물 소화율은 CR20 실험구가 CR40, CR60 및 R80 실험구보다 유의적으로 높았으며, 외관상 지질 소화율은 CR0 실험구가 다른 실험구보다 유의적으로 높았다(P < 0.05). 혈액의 total protein, triglyceride, glucose, aspartate aminotransferase activity, alanine aminotransferase activity, albumin, lactate dehydrogenase, alkaline phosphatase 및 total bilirubin의 경우 모든 실험구간에서 유의적인 차이가 없었다(P > 0.05). 하지만 total cholesterol 및 high-density lipoprotein cholesterol의 경우 쌍별귀뚜라미의 첨가량에 따라서 감소하는 경향을 나타내었다(P < 0.05). 전어체 및 근육 조지질 함량은 사료내 쌍별귀뚜라미의 첨가량에 따라서 유의적으로 감소하는 경향을 나타내었다. 근육 n-3 고도 불포화지방산의 함량은 쌍별귀뚜라미 첨가량에 따라서 감소하는 경향을 나타내었다. 반면에 항산화 효소 활성의 경우 쌍별귀뚜라미 첨가량에 따라서 증가하였지만 어분 60% 이상 대체시에는 감소하는 경향을 나타내었다. 따라서 넙치 사료내 어분대체원으로서 쌍별귀뚜라미는 성장 및 면역에 부정적인 영향 없이 사용이 가능할 것으로 판단된다. 2. 넙치 치어 사료내 어분대체원으로서 갈색거저리의 이용성 본 연구는 어분 대체원으로 갈색거저리를 첨가한 배합 사료가 넙치 치어의 성장, 체조성 및 면역에 미치는 영향을 조사하기 위해 수행되었다. 실험사료의 주 단백질원으로 정어리어분 및 멸치어분을 사용하였으며, 지질원으로 어유 및 대두유를 사용하였다. 그리고 사료 내 갈색거저리 첨가에 따른 넙치의 반응을 알아보기 위해 대조사료(MW0)의 조성물 중 어분, 소맥분 및 대두유의 첨가비를 감소시키면서 갈색거저리를 각각 13, 26, 39 및 52% 첨가하여 총 5종류의 실험사료를 설계하였다. 사육실험은 평균 33.5±0.01 g 전후의 넙치를 선별하여 150L 원형수조에 각각 20마리씩 3반복으로 수용하여 8주간 수행하였다. 넙치 치어의 일간성장율은 갈색거저리 첨가량에 따라서 감소하는 경향을 나타내었다(linear and quadratic, P < 0.05). 외관상 단백질소화율은 MW0, MW20 및 MW40 실험구가 MW60 및 MW80 실험구보다 유의적으로 높았다(P < 0.05). 혈액 total choresterol 함량의 경우 갈색거저리 첨가량에 따라서 감소하는 경향을 나타내었다(linear, P < 0.05). 등근육 조지질 함량은 MW0 실험구가 갈색거저리 첨가 실험구보다 유의적으로 높았다(P < 0.05). 등근육 지방산 함량은 사료내 갈색거저리 첨가에 따른 영향을 받았으며, Σn3/Σn6의 경우 MW0 실험구가 MW60 및 MW80 실험구보다 유의적으로 높았다(P < 0.05). 혈청 GPx 함량은 갈색거저리 첨가량에 따라서 증가하는 경향을 나타내었다(linear and quadratic, P < 0.05). 또한 사료내 갈색거저리 첨가에 따른 장내 미생물 군집의 다양성 차이는 나타나지 않았다. 장내 세균 문 수준의 분포는 인정되지 않은 그룹을 포함하여 총 6개의 세균 문이 존재하였으며, 모든 실험구에서 proteobacteria가 가장 우점하였다. 3. 감성돔 치어 사료내 어분대체원으로서 쌍별귀뚜라미의 이용성 본 연구는 어분대체원으로서 쌍별귀뚜라미를 첨가한 배합 사료가 감성돔 치어의 성장, 체조성 및 면역에 미치는 영향을 조사하기 위해 수행되었다. 실험사료의 주 단백질원으로 정어리어분 및 멸치어분을 사용하였으며, 지질원으로 어유 및 대두유를 사용하였다. 그리고 사료 내 쌍별귀뚜라미 첨가에 따른 감성돔의 반응을 알아보기 위해 대조사료(CR0)의 조성물 중 어분, 소맥분, 감자전분, 어유 및 대두유의 첨가비를 감소시키면서 갈색거저리를 각각 8.4, 16.8, 25.2 및 33.6% 첨가하여 총 5종류의 실험사료를 설계하였다. 사육실험은 평균 6.43±0.00 g 전후의 감성돔을 선별하여 300L 원형수조에 각각 35마리씩 3반복으로 수용하여 12주간 수행하였다. 사육실험 종료 후 감성돔의 성장 및 생존율은 유의적인 차이가 없었다. 감성돔의 일일사료섭취율은 CR0 실험구가 CR60 실험구보다 유의적으로 높았다(P < 0.05). 혈장 조성의 경우 쌍별귀뚜라미 첨가에 따른 유의적인 차이가 없었다(P > 0.05). 근육 조지질 함량은 쌍별귀뚜라미 첨가량에 따라 감소하는 경향을 나타내었다(linear and quadratic, P < 0.05). 근육 Σn3 / Σn6 비율은 쌍별귀뚜라미 첨가에 따라 감소하는 경향을 나타내었다(linear and quadratic, P < 0.05). 혈청 lysozyme 활성의 경우 쌍별귀뚜라미 첨가에 따라서 증가하는 경향을 나타내었다(linear and quadratic, P < 0.05). 4. 감성돔 치어 사료내 어분대체원으로서 갈색거저리의 이용성 본 연구는 어분대체원으로서 갈색거저리를 첨가한 배합 사료가 감성돔 치어의 성장, 체조성 및 면역에 미치는 영향을 조사하기 위해 수행되었다. 실험사료의 주 단백질원으로 정어리어분 및 멸치어분을 사용하였으며, 지질원으로 어유 및 대두유를 사용하였다. 그리고 사료 내 쌍별귀뚜라미 첨가에 따른 감성돔의 반응을 알아보기 위해 대조사료(MW0)의 조성물 중 어분, 소맥분, 감자전분, 어유 및 대두유의 첨가비를 감소시키면서 갈색거저리를 각각 8.4, 16.8, 25.2 및 33.6% 첨가하여 총 5종류의 실험사료를 설계하였다. 사육실험은 평균 6.43±0.00 g 전후의 감성돔을 선별하여 300L 원형수조에 각각 35마리씩 3반복으로 수용하여 12주간 수행하였다. 사육실험 종료 후 감성돔의 성장 및 생존율은 유의적인 차이가 없었다. 감성돔의 일일사료섭취율은 갈색거저리 첨가량에 따라서 감소하는 경향을 나타내었다(Linear, P < 0.05). 혈장 조성의 경우 갈색거저리 첨가에 따른 유의적인 차이가 없었다(P > 0.05). 근육 조지질 함량은 갈색거저리 첨가량에 따라 감소하는 경향을 나타내었다(P < 0.05). 감성돔 근육 내 n-6 지방산 함량은 갈색거저리 첨가량에 따라 감소하는 경향을 나타내었다(linear and quadratic, P < 0.05). 혈청 lysozyme 활성의 경우 쌍별귀뚜라미 첨가에 따라서 증가하는 경향을 나타내었다(linear and quadratic, P < 0.05). 간 조직의 catalase 및 glutathione peroxidase 발현량은 갈색거저리 첨가에 따라서 유의적으로 증가하였다(P < 0.05).
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CHAPTER 3 Mealworm, Tenebrio molitor, as a feed ingredient for juvenile olive flounder, Paralichthys olivaceus Abstract An 8-week feeding trial was designed to evaluate the potential of mealworm (Tenebrio molitor) as a locally available unconventional source of protein in a practical diet for olive flounder (Paralichthys olivaceus). Three hundred juveniles (initial weight, 33.5 ± 0.01 g) were randomly distributed into five dietary groups in triplicate (20 fish per tank), and each group was hand-fed one of the experimental diets containing a graded level of a mealworm meal (MW) replacing 0, 20, 40, 60, and 80% of fish meal (FM) protein (designated as MW0, MW20, MW40, MW60, and MW80). The specific growth rates of juvenile flounders revealed a decreasing trend with increasing dietary MW levels. However, feed utilization efficiency tended to improve with the increasing percentage of dietary FM replacement up to 40% but gradually decreased in higher replacement levels. The results displayed a linear and quadratic negative effect of dietary MW inclusion level on nutrient digestibility. A significant negative linear trend was detected between dietary MW content and plasma total cholesterol. Except for lipid content, which displayed a significant decreasing trend as the dietary MW content increased, no other changes were evident in the fillet proximate composition. Significant positive linear and quadratic trends were observed between fillet C18:1n-9 (oleic acid; OA) and C18:2n-6 (linoleic acid; LA) levels and dietary MW content, where the OA and LA values increased in a dose-dependent manner. The fillet Σn3 / Σn6 ratio and atherogenic index demonstrated significant decreasing trends with dietary inclusion of MW while the fillet thrombogenic index and hypocholesterolaemic/hypercholesterolaemic fatty acid ratio exhibited significant positive trends with increasing values. Myeloperoxidase activity declined with replacement of up to 40% FM protein but thereafter gradually increased at higher replacement levels. Glutathione peroxidase activity generally was enhanced with increasing dietary MW inclusion. Dietary MW inclusion had no significant impact on the bacterial community of the juvenile flounder’s hindgut. Our findings could promote the utilization of MW in olive flounder feed, which may not only reduce dependence on high priced and finite feed ingredients, but also enhance farmed fish health status.
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CHAPTER 2 Nutritional evaluation of cricket, Gryllus bimaculatus, meal as fish meal substitute for olive flounder, Paralichthys olivaceus, juveniles Abstract An 8-week feeding trial was designed to evaluate the potential of cricket meal, Gryllus bimaculatus, as a locally available unconventional source of protein in a practical diet for olive flounder. Three hundred juveniles (Initial weight, 33.5 ± 0.01 g) were randomly distributed into five dietary groups in triplicates (20 fish per tank) and each group was hand-fed one of the experimental diets containing graded level of a cricket meal (CR) replacing 0, 20, 40, 60, and 80% of fish meal (FM) protein (designated as CR0, CR20, CR40, CR60, and CR80). Although replacing more than 40% of dietary FM protein resulted in lower growth rates and feed utilization efficiency, juvenile flounders fed the highest level of dietary CR (CR80) still performed as well as control group (CR0). Replacing more than 20% of FM with CR markedly reduced apparent nutrient digestibility in fish. Plasma total cholesterol concentration showed decreasing trend with increasing inclusion levels of CR in the diet. Whole body and fillet lipid content decreased with increasing level of dietary CR. There was a remarkable enhancement in the levels of C18 fatty acids and reduction in the levels of n-3 highly unsaturated fatty acids in the fillets of fish fed the CR diets, which became more prominent with greater levels of dietary FM replacement. Antioxidant enzyme activities tended to increase with increasing dietary CR levels but gradually decreased when the replacement level exceeded 60%. Overall, the efficiency of CR as a promising substitute for FM in the flounder diets has been confirmed not only in relation to growth rates, but also in terms of immunopotency.
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CHAPTER 4 Nutritional evaluation of cricket, Gryllus bimaculatus, meal as fish meal substitute for black porgy, Acanthopagrus schlegelii, juveniles Abstract A 12-week feeding trial was designed to evaluate the potential use of cricket meal, Gryllus bimaculatus, as a locally available unconventional source of protein in a practical diet for black porgy. Five hundred twenty five healthy juveniles (Initial weight; 6.43 ± 0.00 g) were randomly distributed into five dietary groups in triplicate (35 fish per tank) and each group was hand-fed with one of the experimental diets containing graded level of a cricket meal (CR) replacing 0, 15, 30, 45, and 60% of fish meal (FM) protein (designated as the CR0, CR15, CR30, CR45, and CR60 diets, respectively). Although replacing 60% of dietary FM protein resulted in lower daily feed intake, juvenile porgies fed the CR60 performed as well as CR0 diet. A significantly negative linear trend was detected between fillet lipid content and increasing inclusion levels of CR in diets. There was a notable reduction in the n-3 polyunsaturated fatty acids in the fillets of fish fed the CR diets substituting CR for FM, which became more prominent with greater levels of dietary FM replacement. Serum myeloperoxidase activity tended to decrease with increasing dietary CR levels, but increased when the replacement level exceeded 45%. Serum lysozyme activity improved with increasing inclusion levels of CR in diets. Overall, the efficiency of CR as a promising substitute for FM in the black porgy diets has been confirmed not only in relation to growth rates, but also in terms of immunopotency. Our findings could promote the utilization of CR in black porgy feed, which may not only reduce dependence on high priced and finite feed ingredients, but also enhance farmed fish health status.
more초록/요약 도움말
CHAPTER 5 Mealworm, Tenebrio molitor, as a feed ingredient for black porgy, Acanthopagrus schlegelii, juveniles Abstract A 12-week feeding trial was designed to evaluate the potential use of mealworm, Tenebrio molitor, as a locally available unconventional alternative source of protein for fish meal (FM) in a practical diet for black porgy. Five hundred twenty five healthy juveniles (initial weight; 6.43 ± 0.00 g) were randomly distributed into five dietary groups in triplicate (35 fish per tank) and each group was hand-fed with one of the experimental diets containing graded level of a mealworm meal (MW) replacing 0, 15, 30, 45, and 60% of FM protein designated as MW0, MW15, MW30, MW45, and MW60 diets, respectively. Although replacing 60% of dietary FM protein resulted in lower daily feed intake, juvenile porgies fed the highest level of dietary MW (MW60) still performed as well as control group (MW0). A significant negative trend was detected between fillet lipid content and increasing inclusion levels of MW in diets. There was a notable reduction in the n-3 polyunsaturated fatty acids in the fillets of fish fed the MW diets, which became more prominent with greater levels of dietary FM replacement. Serum lysozyme activity improved with increasing inclusion levels of MW in the diet. Furthermore, expression of the antioxidant enzyme-related genes including glutathione peroxidase (GPx) and catalase (CAT) were markedly upregulated in liver of the fish fed diets replacing FM with MW when compared to fish fed the MW0 diet. Overall, the efficacy of MW as a promising alternative dietary protein source for juvenile black porgy has been proved not only in relation to weight gain and efficiency of feed utilization, but also the view point of health-promoting effects in fish.
more초록/요약 도움말
Summary The nutritive needs of monogastric species, particularly fish, include a high quality and quantity of protein in the diet. Fish meal (FM) is a rich source of protein; however, due to high cost and dependency on its import, there is an increased demand for the alternative and easily available protein sources. Recently, there is an increased interest in the utilization of insect meals as potential protein source in aquafeed. However, there is limited information on their nutritional value as potential dietary ingredients in diets for olive flounder and black porgy which are important marine finfish species cultured in South Korea. Therefore, a series of feeding trials were designed to provide insights into the feasibility of replacing FM by two different insect meals, including cricket and mealworm meals, in diets for both fish species. We believe that the findings of these reseachs will encourage feed manufactures to utilize insect meals more efficiently in generating low-cost and sustainable aquafeed, not only to reduce their dependence on high priced and finite feed ingredients, but also to enhance production performance and health status of farmed seafood. The short justification of each chapter is as follows: Chapter 2 is about the potential of cricket meal (CR) as a locally available unconventional source of protein in a practical diet for juvenile olive flounder. For this, flounders were fed with diets containing graded level of a cricket meal (CR) replacing 0, 20, 40, 60, and 80% of FM protein (designated as CR0, CR20, CR40, CR60, and CR80). In this study, we demonstrated that although flounder juveniles are able to tolerate up to 80% substitution of FM protein with CR, highly beneficial n-3 long-chain polyunsaturated fatty acid (LC-PUFA) content of their fillet remarkably reduced at the highest levels of CR inclusion (80%) which is not an ideal trait from the consumer’s perspective. In addition, chapter 2 documents that dietary inclusion of the CR could notably enhance fish antioxidant enzyme activities when compared to those groups offered the CR free diets (CR0). Chapter 3 examines a localy available mealworm meal (MW) as a nutritious and sustainable source of protein in a practical diet for olive flounder juveniles. For this, flounders were fed with diets containing graded level of MW replacing 0, 20, 40, 60, and 80% of FM protein (designated as MW0, MW20, MW40, MW60, and MW80). While flounder’s growth negatively correlated with dietary MW inclusion, their feed utilization efficiency tended to improve up to 40% of FM replacement, and then gradually declined when dietary FM replacement further increased to 80%. Dietary MW inclusion was also found to negatively affecting the fish nutrient digestibility and highly beneficial n-3 LC-PUFA in their fillet. Additionally, FM replacement with MW was found to promote an immunostimulatory effect in juvenile flounders. The results in the current chapter may offer convincing evidence that FM protein could be substituted by MW without any adverse impact on juvenile olive flounder gut microbial community, when compared to the fish fed the MW-free control diet. The aim of the study presented in chapter 4 was to describe the supplemental effects of the CR in a practical diet, and to compare its efficiency with that of the CR-free diet for juvenile black porgy. For this, fish were fed with diets containing graded level of CR replacing 0, 15, 30, 45, and 60% of FM protein (designated as CR0, CR15, CR30, CR45, and CR60). This chapter indicated that up to 60% of the FM protein could be replaced by dietary inclusion of the tested CR without any significant negative effects on fish performance. There was a notable reduction in the n-3 polyunsaturated fatty acids in the fillets of fish fed the CR diets, which became more prominent with greater levels of dietary FM replacement. It was also demonstrated that CR inclusion can positively influence fish non-specific immune response, encouraging its dietary utilization as an efficient substitute for FM not only in relation to growth rates, but also in terms of immunopotency. In chapter 5, we discuss the feasibility of dietary utilization of the MW as an alternative source of protein in a practical diet for black porgy juveniles. This chapter indicates that up to 60% of the dietary FM protein could be replaced by the MW without causing any remarkable negative impact on fish performance. Perhaps the only drawback of using MW in black porgy diet was that fillet of fish fed diets containing MW, at the expense of FM, had lower levels of n-3 PUFAs which is not an ideal trait, from the nutritional point of view, for human consumption. It was also indicated that the dietary inclusion of the MW in diets for black porgy could notably enhance their hepatic glutathione peroxidase (GPx) and catalase (CAT) mRNA expression, which are fundamentally involved in cellular redox homeostasis. These findings suggested that MW might prove to be a promising alternative to FM in practical diets for juvenile black porgy without having any adverse consequences for the health and performance of the fish.
more목차 도움말
CONTENTS
page
LISTS OF TABLES i
LISTS OF FIGURES ii
Summary 1
1. Introduction 4
1.1. Fish meal as an excellent feed ingredient in aquafeed 4
1.2. Insect meals as an alternative protein source in aquafeed 6
1.3. Cricket (Gryllus bimaculatus) meal as a protein source in aquafeed 7
1.4. Mealworm (Tenebrio molitor) meal as a protein source in aquafeed 8
1.5. Potential applications of insect meals in aquafeed for olive flounder (Paralichthys olivaceus) 9
1.6. Potential applications of insect meals in aquafeed for black porgy (Acanthopagrus schlegelii) 9
2. Nutritional evaluation of cricket (Gryllus bimaculatus) meal as fish meal substitute for olive flounder (Paralichthys olivaceus) juveniles 11
2.1. Abstract 11
2.2. Material and Methods 12
2.3. Results 23
2.4. Discussion 38
3. Mealworm (Tenebrio molitor) as a feed ingredient for juvenile olive flounder (Paralichthys olivaceus) 43
3.1. Abstract 43
3.2. Material and Methods 45
3.3. Results 56
3.4. Discussion 74
4. Nutritional evaluation of cricket (Gryllus bimaculatus) meal as fish meal substitute for black porgy (Acanthopagrus schlegelii) juveniles 80
4.1. Abstract 80
4.2. Material and Methods 81
4.3. Results 85
4.4. Discussion 97
5. Mealworm (Tenebrio molitor) as a feed ingredient for black porgy (Acanthopagrus schlegelii) juveniles 102
5.1. Abstract 102
5.2. Material and Methods 103
5.3. Results 109
5.4. Discussion 122
Conclusion 128
국문요약 129
Acknowledgement 133
List of References 134
