Ảnh hưởng của một số chiết xuất thảo dược lên các chỉ tiêu sinh lý cá tra Pangasianodon hypophthalmus

MINISTRY OF EDUCATION AND TRAINING CAN THO UNIVERSITY PHAM NGOC NHU EFFECTS OF DIETARY SUPPLEMENTATION WITH SELECTED PLANT EXTRACTS ON THE PHYSIOLOGICAL PARAMETERS OF STRIPED CATFISH (Pangasianodon hypophthalmus) DOCTORAL DISSERTATION MAJOR: AQUACULTURE (9620301) 2023 TABLE OF CONTENTS DATA SHEET. iii LIST OF FIGURES .ix LIST OF TABLES .xi LIST OF ABBREVIATION .2 The objectives of the dissertation. Error! Bookmark not defined.3 The main contents of the dissertation .4 The hypotheses of the dissertation .5 New findings of the dissertation .6 Significant contributions of the dissertation .1 The status and importance of aquaculture and fisheries.2 Climate changes and impacts on aquaculture and fisheries .3 Mechanism of stress on fish .4 Effects of environmental factors on fish .1 Effect of temperature on fish .2 Effects of salinity on fish .3 Effects of nitrite on fish .5 Effects of plant extracts on fish .1 Effects of plant extracts on hematological parameters of fish .2 Effects of plant extracts on digestive enzymes activities and growth of fish .3 The effects of plant extracts on oxidative stress of fishes .4 Effects of plant extracts on striped catfish (P. 62 a) Effects of Mimosa pudica on aquaculture species.

67 b) Effects of Psidium guajava on aquaculture species. 67 c) Effect of Phyllanthus amarus on fish. 71 d) Effect of Euphorbia hirta on fish. 77 e) Effect of Azadirachta indica on fish.

81 EFFECTS OF PLANT EXTRACTS ON SELECTED HAEMATOLOGICAL PARAMETERS, DIGESTIVE ENZYMES, AND GROWTH PERFORMANCE v OF STRIPED CATFISH, Pangasianodon hypophthalmus (Sauvage, 1878) FINGERLINGS .2 Material and Method .1 Plant extract and feed preparation .2 Experimental fish acclimation, facilities, and feeding management .3 Haematological and biochemical parameters .4 Digestive enzyme activities .5 Growth performance and survival rate .1 Effects of plant extracts on haematological parameters .2 Effects of plant extract on digestive enzyme activities .3 Effects of plant extracts on growth performance and the survival rate. 97 EFFECTS OF Psidium guajava AND Phyllanthus amarus EXTRACTS ON HAEMATOLOGICAL PARAMETERS, DIGESTIVE ENZYMES ACTIVITIES OXIDATIVE STRESS, AND GROWTH OF STRIPED CATFISH, Pangasianodon hypophthalmus FINGERLINGS EXPOSED TO HIGH-TEMPERATURE STRESS. Materials and methods .1 Plant extracts and diet preparation .2 Experimental fish acclimation, facilities, and feeding trial .3 Haematological and biochemical parameters .4 Oxidative stress assays .1 Effect of plant extract on P. hypophthalmus haematological parameters .2 Effect of plant extract on digestive enzyme activities .3 Effect of plant extract on growth performance of P.4 Effect of plant extract on P.

hypophthalmus oxidative stress .123 vi EFFECTS OF GUAVA (Psidium guajava L.) AND BHUMI AMLA (Phyllanthus amarus Chum et Thonn.) EXTRACTS ON HAEMATOLOGICAL PARAMETERS, DIGESTIVE ENZYMES ACTIVITIES OXIDATIVE STRESS, AND GROWTH OF STRIPED CATFISH, Pangasianodon hypophthalmus FINGERLINGS EXPOSED TO SUBLETHAL SALINITIES .2 Material and Method .1 Plant extract and feed preparation .2 Experimental fish acclimation, facilities, and feeding management .3 Haematological and biochemical parameters .4 Digestive enzyme activities .5 Oxidative stress biomarkers .6 Growth performance and survival rate .1 Effects of plant extracts on haematological parameters .2 Effects of plant extracts on digestive enzyme activities.3 Effects of plant extracts on oxidative stress .4 Effects of plant extracts on growth performance and the survival rate .126 EFFECTS OF Psidium guajava AND Phyllanthus amarus EXTRACTS ON HAEMATOLOGICAL PARAMETERS, DIGESTIVE ENZYMES ACTIVITIES OXIDATIVE STRESS, AND GROWTH OF STRIPED CATFISH, Pangasianodon hypophthalmus FINGERLINGS EXPOSED TO NITRITE-INDUCED TOXICITY .2 Material and Method .1 Plant extract and feed preparation .2 Experimental fish acclimation, facilities, and feeding management .3 Haematological and biochemical parameters .4 Digestive enzyme activities .5 Oxidative stress biomarkers .6 Growth performance and survival rate .1 Effects of plant extracts on haematological parameters .2 Effects of plant extracts on digestive enzyme activities.3 Effects of plant extracts on oxidative stress .4 Effects of plant extracts on growth performance and the survival rate .1 Effects of five selected extracts on physiology parameters and stress mitigation of P.2 Effects of Psidium guajava and Phyllanthus amarus extracts on physiology parameters and stress mitigation of P. hypophthalmus fingerlings exposed to high- temperature stress .3 Effects of Psidium guajava and Phyllanthus amarus extracts on physiology parameters and stress mitigation of P. hypophthalmus fingerlings exposed to sublethal salinities .4 Effects of Psidium guajava and Phyllanthus amarus extracts on physiology parameters and stress mitigation of P. hypophthalmus fingerlings exposed to nitrite- induced toxicity .153 CONCLUSIONS AND RECOMMENDATIONS .2 Recommendations for further studies.

1 List of protocols of analysis used in the studies. 1 viii LIST OF FIGURES Figure 2.1 Total fisheries and aquaculture production 2020 (FAO, 2022).2 World fisheries and aquaculture production, utilization and trade .3 World production of striped catfish (thousand tons) (FAO, 2022).4 Farming area and production of striped catfish in Mekong Delta (2015-2021; VASEP, 2022) .5 Export value of striped catfish in the period 2015-2021 (VASEP, 2022). Stress response in response to temperature elevation. (Igwe et al., 2016); Phyllanthus amarus (Abeng, 2017); Mimosa pudica (Goyal, 2014); Psidium guajava (Deepa et al., 2017); Azadirachta indica (Neelakantan et al.8 Antioxidant activities of different A.

indica extracts and their bioactive compounds.1 Plasma glucose concentrations (mg/100 mL) of P. hypophthalmus fed extract-based diets in a 60-day experiment.2 The pepsin (A) and gastric amylase (B) activities (U min/mg protein) of P. hypophthalmus fed extract-based diets in a 60-day experiment.3 The intestinal amylase (A), trypsin (B) and chymotrypsin (C) activities (U min/mg protein) of P. hypophthalmus fed extract-based diets in a 60-day experiment.1 Pepsin (A) and gastric amylase (B) activities of P.

hypophthalmus fingerlings under various temperatures for 42 days.2 Intestinal amylase (A), trypsin (B), and chymotrypsin (C) activities of P. hypophthalmus fingerlings under various temperatures for 42 days.4 LPO-gill (A) and LPO-liver (B) of P. hypophthalmus fingerlings under various temperatures in 42 days.3 Growth performance (WG, DWG, and SGR) and survival rates of P. hypophthalmus fingerlings under various temperatures for 42 days.5 LPO-brain (A) and LPO-muscle (B) of P.

hypophthalmus fingerlings under various temperatures in 42 days.7 CAT-gill (A) and CAT-liver (B) of P. hypophthalmus fingerlings under various temperatures in 42 days. hypophthalmus fingerlings under various salinity in 14 days. Asterisk (*) present significant differences (p<0.05) among salinity levels whereas different lowercase letters (a, b) indicate significant differences among feed groups (p<0.

hypophthalmus fingerlings under various. 3 Glucose concentration (A), osmolality (B) of P. hypophthalmus fingerlings under various salinity in 14 days. Asterisk (*) presents significant differences (p<0.05) among salinity levels .4 LPO- liver (A), LPO-gill (B), LPO-muscle (C) and LPO-brain (D) of P.

hypophthalmus fingerlings under various salinity in 42 days. Asterisk (*) present significant differences (p<0.05) among salinity levels whereas different lowercase letters (a, b) indicate significant differences among feed groups (p<0. 5 CAT-liver (A), CAT-gill (B), CAT- brain (C) and CAT- muscle (D) of P. hypophthalmus fingerlings under various salinity in 42 days.

Asterisk (*) present significant differences (p<0.05) among salinity levels whereas different lowercase letters (a, b) indicate significant differences among feed groups (p<0.1 RBCs, Hb, and Hct of P. hypophthalmus fingerlings exposed to different nitrite concentrations for 14 days. hypophthalmus fingerlings exposed to different nitrite concentrations for 14 days.3 Pepsin and gastric amylase activities of P. hypophthalmus fingerlings exposed to different nitrite concentrations for 42 days.4 Intestinal amylase, trypsin, and chymotrypsin activities of P.

hypophthalmus fingerlings exposed to different nitrite concentrations for 42 days.5 DWG, WG, and survival rate of P. hypophthalmus fingerlings exposed to different nitrite concentrations for 42 days. 142 x LIST OF TABLES Table 2.1 Effects of salinity on fish’s physiology under culture .2 Effects of some herbal extracts on hematological parameters of fish under culture .3 Effects of some herbal extracts on digestive enzyme activities of fish under culture .4 Main oxygen reactive species and their performance Garcez et al.5 Antioxidant defense system of cell enzymes and biological mechanism (Shi and Noguchi, (2001).6 Main groups of plant compounds with antimicrobial activity (Cowan, 1999) .7 Effects of some herbal extracts on oxidative stress indices of fish under culture .8 Lists of plant extract (from World Flora Online, https://wfoplantlist.org/plant-list/) .9 Identification information of five plant extracts used in the experiment (Dao et al.10 Phytochemical screening of Mimosa pudica .11 Bioactive metabolites of Mimosa pudica (Rizwan et al.12 Phytochemical screening of Psidium guajava. 13 Phytochemical screening of different species of Psidium guajava .14 Bioactive metabolites of Psidium guajava .15 Bioactive metabolites of P.

amarus (Patel et al.16 Phytochemical screening of Euphorbia hirta (Asha et al.1 Experimental feed ingredients and formulation .2 Haematological parameters of P. hypophthalmus fed extract-based diets in a 60-day experiment .3 Growth performance of P. hypophthalmus fed plant extract-based diets in a 60-day experiment .1 Experimental feed ingredients and formulation. hypophthalmus fingerlings under various temperatures in.

hypophthalmus fingerlings under various temperatures in 14 days. hypophthalmus fingerlings under various temperatures in 14 days. hypophthalmus fingerlings under various temperatures in 14 days. hypophthalmus fingerlings under various temperatures in 14 days.

hypophthalmus fingerlings under various temperatures in 14 days .8 Glucose concentration (mg/100 mL) of P. hypophthalmus fingerlings under various temperatures in 14 days. hypophthalmus fingerlings exposed to different salinities for 42 days.2 Gastric amylase of P. hypophthalmus fingerlings exposed to different salinities for 42 days.

hypophthalmus fingerlings exposed to different salinities for 42 days.1 Glucose concentration (mg/100 mL) of P. hypophthalmus fingerlings exposed to different nitrite concentrations for 14 days ……………… 133 Table 6.2 LPO-gill (nmol MDA/g) of P. hypophthalmus fingerlings exposed to different nitrite concentrations for 42 days .3 LPO-brain (nmol MDA/g) of P. hypophthalmus exposed to different nitrite concentrations for 42 days .4 LPO-muscle (nmol MDA/g) of P.

hypophthalmus exposed to different nitrite concentrations for 42 days .5 CAT-liver (U/min/mg protein) of P. hypophthalmus exposed to different nitrite concentrations for 42 days .6 CAT-brain (U/min/mg protein) of P. hypophthalmus exposed to different nitrite concentrations for 42 days .7 CAT-muscle (U/min/mg protein) of P. hypophthalmus exposed to different nitrite concentrations for 42 days .1 Physiological parameters of striped catfish fed extract-based diets at 31°C and 35°C compared to 27°C .2 Physiological parameters of striped catfish fed extract-based diets at 10‰ and 20‰ compared to 0‰ .3 Physiological parameters of striped catfish fed extract-based diets at 0.8 mM compared 0 mM .154 xii LIST OF ABBREVIATION CAT Catalase DWG Daily weight gain FAO Food and Agriculture Organization FCR Feed conversion ratio Fig.

Figure GW Gained weight Hb Haemoglobin Hct Haematocrit LC50 96 h Lethal concentration in 96 hours LPO Lipid peroxidation MCH Mean corpuscular haemoglobin MCHC Mean corpuscular haemoglobin concentration MCV Mean corpuscular volume MRD Mekong River Delta Mt Metric ton MIC Minimum inhibitory concentration NO2- Nitrite RBCs Red blood cells SEM Standard error of mean SGR Specific growth rate SR Survival rate WG Weight gain WBCs Number of white blood cells (leukocytes) Ai Azadirachta indica Eh Euphorbia hirta Mp Mimosa pudica Pa Phyllanthus amarus Pg Psidium guajava xiii ABSTRACT Striped catfish (Pangasianodon hypophthalmus) is an important species of aquaculture in the Mekong River Delta (MRD) of Vietnam. However, it has been constrained by several obstacles, among them which are climate change and diseases. Plant extracts as a dietary supplement is regarded as the easiest and most efficient strategy to improve antioxidant activity while contributing to the stress mitigation. The study aimed to evaluate the effect of selected plant extracts medicated in feed on physiological haematological parameters, digestive enzyme activities and growth and stress responses of striped catfish exposed to environmental stressors (salinity, temperature and NO2-).

The final aim would be a selection of plant extracts that have a positive effect on fish to reduce the use of antibiotics and avoid water environmental pollution. This doctoral dissertation was, therefore structured into four separate experiments. First, five plant extracts, 0.4% or 2% Euphorbia hirta (Eh), 0.2% or 1% Phyllanthus amarus (Pa), 0.4% or 2% Mimosa pudica (Mp), 0.2% or 1% Psidium guajava (Pg), and 0.4% or 2% Azadirachta indica (Ai), were investigated on haematology, digestive enzyme activities and growth throughout the duration of 60 days. These extracts were identified based on the promising and applicable findings regarding the immunity and antioxidant capacity of striped catfish reported (Nhu et al., 2019; Dao et al.