Positive effects of Andrographis paniculata extract on growth performance, haematology, serum biochemistry, organosomatic indices and resistance against Aeromonas hydrophila in hybrid catfish (Clarias macrocephalus × Clarias gariepinus)

Figures & data

Table 1. Preliminary phytochemical screening methods for Andrographis paniculata.

Phytochemicals	Tests	Observations	References
Flavonoids	Shinoda	Appearance of crimson red colour	Shaikh and Patil 2020
Alkaloids	Dragendroff	Formation of reddish-brown precipitate	Pant et al. 2017
	Mayer	Formation of cream precipitate
Anthraquinones	Modified Borntrager	Appearance of pink-red colour	Shaikh and Patil 2020
Coumarins	Sodium hydroxide paper	Yellow fluorescence under UV light	Shaikh and Patil 2020
Tannins/Phenolics	Ferric chloride	Appearance of green to dark blue or black colour	Pant et al. 2017
Saponins	Foam test	The persistence of honeycomb froth lasting for longer than half an hour, and still existing after the addition of acid solution	Shaikh and Patil 2020
Diterpenes	Copper acetate test	Appearance of emerald-green colour	Shaikh and Patil 2020
Steroids	Liebermann-Burchard	Change of colour from violet to blue or green	Shaikh and Patil 2020
Cardiac glycosides	Liebermann-Burchard	Steroid nuclei; Change of colour from violet to a blue-green colour	Shaikh and Patil 2020
	Kedde	Five-membered unsaturated lactone rings; Formation of reddish-brown junction between two liquid layers with a greenish appearance in the upper layer
	Keller-Kiliani	Deoxy sugar; Appearance of purplish-pink colour
Cyanogenic glycosides	Grignard	Appearance of red-brown colour	Hartanti and Cahyani 2020
Sugars/Carbohydrates	Molisch	Appearance of purple ring	Shaikh and Patil 2020

Table 2. Preliminary phytochemical constituents of 95% ethanol extract from Andrographis paniculata extract.

Phytochemicals results
Flavonoids	−
Alkaloids	−
Anthraquinones	−
Coumarins	+
Tannins/Phenolics	+
Saponins	−
Terpenoids	+
Lactones (5-membered unsaturated)	+
Steroids	+
Diterpenes	+
Cardiac glycosides	−
Cyanogenic glycosides	−
Sugars/Carbohydrates	+

+ Note: present, − absent.

Figure 1. Growth indices such as weight gain: WG (a), average daily gain: ADG (b), specific growth rate: SGR (c), survival rate: SR (d), feed conversion rate: FCR (e) and protein efficiency ratio: PER (f) of hybrid catfish, Clarias macrocephalus × C. gariepinus, which were fed different levels of Andrographis paniculata. Different letters indicate significant differences (p < 0.05). T1 (0.00 g/kg), T2 (0.2 g/kg), T3 (0.4 g/kg) and T4 (0.6 g/kg).

Figure 2. The hematological indices of hybrid catfish, Clarias macrocephalus × C. gariepinus, which were fed different levels of Andrographis paniculata extract for 30, 60, and 90 days: total white blood cells, WBC (a); total red blood cells, RBC (b); haematocrit, Hct (c); Significant differences (p < 0.05) are indicated by different letters.

Figure 3. Effect of different levels of Andrographis paniculata extract on activities of for analysis. Serum indices, activities of the alanine transaminase ALT (a), aspartate transaminase AST (b), creatinine (c), Alkaline phosphatase (d), glucose (e), total protein (f), albumin (g), globulin (h), Triglycerides (i), cholesterol (j), low density Lipoprotein, LDL (k) and high-density Lipoprotein, HDL (l) in hybrid catfish, Clarias macrocephalus × C. gariepinus. Values are mean ± SD and different letters indicate difference (p < 0.05). T1 (0.00 g/kg), T2 (0.2 g/kg), T3 (0.4 g/kg) and T4 (0.6 g/kg).

Figure 4. Effect of different levels of Andrographis paniculata extract on activities of for analysis lysozyme activity (m) in hybrid catfish, Clarias macrocephalus × C. gariepinus. Values are mean ± SD and different letters indicate difference (p < 0.05). T1 (0.00 g/kg), T2 (0.2 g k), T3 (0.4 g /kg) and T4 (0.6 g/kg).

Table 3. Organosomatic indices of hybrid catfish, Clarias macrocephalus × C. gariepinus were fed with different concentrations of Andrographis paniculata extract for 90 days.

Items	T1	T2	T3	T4
SSI	1.80 ± 0.01	2.20 ± 0.003	2.12 ± 0.01	1.96 ± 0.005
HIS	1.74 ± 0.12	1.77 ± 0.001	1.77 ± 0.02	1.93 ± 0.09
KI	0.47 ± 0.04	0.53 ± 0.02	0.47 ± 0.02	0.55 ± 0.02
ISI	1.80 ± 1.21	2.20 ± 0.09	2.12 ± 1.01	1.96 ± 1.22

Note: HIS, hepatosomatic index; SSI, spleen somatic index; KI, kidney index; ISI, intestinal somatic index. Data represent mean ± SD. Treatment groups: T1 (0.00 g/kg), T2 (0.2 g/kg), T3 (0.4 g/kg) and T4 (0.6 g/kg).

Figure 5. Cumulative mortality (%) of hybrid catfish, Clarias macrocephalus × C. gariepinus, fed with the four different concentrations of Andrographis paniculata extract against Aeromonas hydrophila for 14 days. T1 (0.00 g /kg), T2 (0.2 g/ kg), T3 (0.4 g/ kg) and T4 (0.6 g /kg).

Table 4. Blood parameters and lysozyme activity of hybrid catfish, Clarias macrocephalus × C. gariepinus after Aeromonas hydrophila injections at 10⁹ CFU/ml for 14 days.

Items	T1	T2	T3	T4
RBC (10^9 cell/mm^3)	2.58 ± 0.15^b	2.79 ± 0.16^ab	2.68 ± 0.11^ab	3.19 ± 0.09^a
WBC (10^7 cell/mm^3)	1.86 ± 0.02^b	2.31 ± 0.03^a	2.21 ± 0.2^a	2.40 ± 0.01^a
HCT (%)	20.75 ± 0.5^b	24.67 ± 0.19^a	23.34 ± 0.25^ab	25.5 ± 0.33^a
Lysozyme activity (Unit/ml)	20.75 ± 1.2^b	24.67 ± 0.4^a	23.34 ± 0.33^a	25.5 ± 0.88^a

Note: RBC, total red blood cell; WBC, Total white blood cell; HCT, haematocrit percentage. Data represent mean ± SD. Different letters (a–b) in the same row indicate significant differences (p < 0.05). Treatment groups: T1 (0.00 g/kg), T2 (0.2 g/kg), T3 (0.4 g/kg) and T4 (0.6 g/kg).

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