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In vitro evaluation of hydro-ethanolic seed extract of Zanthoxyllum armatum DC to establish its mechanism of action for paralysis of Paramphistomum cervi in continuation with in vivo experimentation in naturally infected goat

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24 mag 2025
INFORMAZIONI SU QUESTO ARTICOLO

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Fig. 1.

Effect of different concentration of HESEZA on glucose uptake in adult P. cervi.
Effect of different concentration of HESEZA on glucose uptake in adult P. cervi.

Fig. 2.

Effect of different concentration of HESEZA on glycogen content in adult P. cervi.
Effect of different concentration of HESEZA on glycogen content in adult P. cervi.

Fig. 3.

Effect of different concentration of HESEZA on lactic acid production in adult P. cervi.
Effect of different concentration of HESEZA on lactic acid production in adult P. cervi.

Fig. 4.

Effect of different concentration of HESEZA on acetylcholinesterase activity in adult P. cervi.
Effect of different concentration of HESEZA on acetylcholinesterase activity in adult P. cervi.

In vivo study of HESEZA (150 and 250 mg/kg b_ wt) on hematological parameters in experimental groups at different days of treatment_

Group Parameter Days of observation
0th 7th 14th 21st 28th
A Hb (g/dl) 7±0.06a 6.97±0.06a 6.97±0.04a 6.9±0.07a 6.85±0.08a
B 7.03±0.11a 7.07±0.13a 7.77±0.12ab 8.02±0.12c 8.28±0.11c
C 7.1±0.1a 7.17±0.08a 7.32±0.08a 7.83±0.1ab 8.57±0.08c
D 6.92±0.06a 7.85±0.04ab 8.05±0.04c 8.6±0.04c 9.25±0.08d
A PCV (%) 20.55±0.34a 20.57±0.36a 20.6±0.34a 20.63±0.34a 20.68±0.3a
B 21.85±0.28a 21.88±0.29a 21.97±0.3ab 22.03±0.28b 22.13±0.29b
C 20.48±0.34a 21.53±0.32a 21.87±0.32a 22.08±0.32b 22.57±0.32b
D 20.32±0.31a 21.78±0.27a 22.05±0.26b 22.77±0.29b 22.93±0.29b
A TLC (103/mm3) 15.78±0.4a 15.86±0.43a 16.24±0.21a 16.17±0.2a 16.09±0.29a
B 15.47±0.25a 14.72±0.35ab 13.68±0.31b 12.33±0.31bc 11.35±0.33c
C 15.68±0.22a 14.1±0.3ab 12.63±0.35bc 11.27±0.37c 9.72±0.32d
D 15.29±0.3a 12.15±0.36bc 10.8±0.27cd 9.38±0.26d 8.9±0.24e
A TEC (106/mm3) 10.48±0.17a 10.28±0.27a 10.34±0.2a 10.41±0.22a 10.38±0.19a
B 9.62±0.18a 10.33±0.15a 11.05±0.19ab 11.8±0.19ab 12.55±0.16b
C 10.2±0.28a 11.23±0.3ab 12.07±0.29b 13.02±0.28bc 13.78±0.26c
D 10.31±0.32a 12.08±0.12b 13.29±0.21bc 13.7±0.16c 13.77±0.19c
A Neutrophil count (%) 51.4±0.36a 51.35±0.33a 51.68±0.38 51.79±0.45 51.58±0.45
B 52.63±1.03a 48.74±0.67b 44.33±0.83bc 40.57±0.77c 36.34±0.92d
C 50.72±1.03a 46.11±0.87b 41.84±0.93c 36.97±1.1d 31.95±1.2e
D 51.01±0.47a 41.28±0.86b 35.59±0.6c 30.76±0.31d 29.25±0.34e
A Eosinophil count (%) 5.62±0.21a 5.63±0.2a 5.66±0.2a 5.69±0.2a 5.7±0.22a
B 5.11±0.13a 4.53±0.14b 3.88±0.1c 3.19±0.11c 2.42±0.12d
C 5.6±0.28a 4.92±0.32b 4.18±0.33b 3.52±0.31c 2.65±0.26d
D 5.51±0.17a 4.6±0.17b 3.83±0.17c 3.05±0.17c 2.25±0.15d
A Monocyte count (%) 4.08±0.13a 4.08±0.12a 4.08±0.13a 4.13±0.14a 4.1±0.13a
B 4.17±0.09a 3.92±0.09a 3.55±0.14b 3.24±0.14b 2.9±0.11bc
C 3.98±0.12a 3.64±0.13ab 3.24±0.11b 2.88±0.1bc 2.36±0.11c
D 4.1±0.12a 3.47±0.14b 3.05±0.11bc 2.62±0.16c 2.29±0.14c
A Lymphocyte count (%) 35.17±1.35a 35.04±1.14a 35.41±1.29a 35.05±1.35a 35.21±1.28a
B 34.17±1.07a 38.21±1.06b 42.26±1.04c 46.8±0.82cd 51.37±0.63e
C 35.62±0.93a 40.11±0.93bc 45.04±0.89c 49.77±0.88d 54.91±0.8e
D 35.74±0.7a 42.72±1.11bc 48.92±1.16c 55.24±1.39d 60.8±1.27e

In vivo study of HESEZA (150 and 250 mg/kg b_ wt) on biochemical parameters in experimental groups at different days of treatment_

Group Parameter Days of observation
0th 7th 14th 21st 28th
A Total serum protein (g/dl) 4.28±0.11a 4.24±0.09a 4.26±0.1a 4.25±0.1a 4.23±0.09a
B 4.27±0.09a 4.67±0.11a 5.03±0.17ab 5.37±0.17ab 5.79±0.16b
C 4.31±0.09a 4.77±0.07a 5.21±0.13ab 5.62±0.13b 6.03±0.14bc
D 4.38±0.08a 4.84±0.13a 5.23±0.14ab 5.65±0.14b 6.19±0.1bc
A Serum albumin (g/dl) 1.82±0.04a 1.81±0.04a 1.81±0.03a 1.81±0.03a 1.82±0.04a
B 1.75±0.03a 1.96±0.05a 2.26±0.08ab 2.64±0.08b 2.94±0.06bc
C 1.83±0.03a 2.21±0.08ab 2.64±0.11b 2.97±0.09bc 3.24±0.09c
D 1.8±0.04a 2.19±0.09ab 2.64±0.08b 2.99±0.08bc 3.26±0.08c
A Serum globulin (g/dl) 2.27±0.05a 2.24±0.05a 2.25±0.05a 2.25±0.06a 2.26±0.06a
B 2.16±0.08a 2.35±0.06ab 2.56±0.07b 2.77±0.06c 2.94±0.05d
C 2.24±0.09a 2.45±0.08ab 2.66±0.07b 2.84±0.06c 3.03±0.06d
D 2.15±0.07a 2.39±0.07ab 2.59±0.08b 2.85±0.06c 3.06±0.06d
A Aspartate aminotransferase level (U/L) 102.65±2.04a 102.95±1.82a 103.32±1.7a 103.32±1.69a 103.7±1.51a
B 100.89±1.51a 93.28±1.69ab 85.82±1.56b 78.27±1.64bc 70.29±1.43c
C 102.99±1.46a 92.93±1.24ab 84.92±1.37b 76.84±1.04bc 67.62±1.14c
D 103.75±2.16a 90.85±1.82ab 82.07±1.95b 71.54±1.22bc 62.49±1.11cd
A Alanine aminotransferase level (U/L) 27.81±0.6a 27.8±0.61a 28.21±0.57a 28.22±0.53a 28.39±0.57a
B 26±0.7a 24.85±0.62b 23.86±0.55c 21.9±0.39d 20.54±0.44d
C 27.23±0.38a 25.5±0.54b 23.71±0.4c 22.12±0.36cd 19.83±0.6de
D 27.38±0.55a 25.47±0.62b 23.02±0.43c 20.57±0.55d 18.96±0.36e
A Alkaline phosphatase level (U/L) 189.72±1.17a 190.44±1.36a 191.76±1.5a 192.05±1.19a 192.5±1.2a
B 189.38±1.42a 177.65±1.31ab 167.06±1.69bc 156.64±1.74cd 145.1±1.47d
C 188.25±1.43a 176.9±1.77ab 164.18±2.28c 152.66±1.84d 140.79±2.2e
D 189.1±2.08a 174.96±1.76b 161.76±1.48c 148.04±1.03d 136.47±1.22e
A Serum iron (µg/dl) 150.26±1.42a 150.32±1.42a 150.29±1.43a 150.26±1.44a 150.25±1.43a
B 152.07±1.54a 155.1±1.5ab 159.68±1.63b 163.59±1.6c 167.52±1.37cd
C 152.52±1.3a 158.22±1.67b 163.05±1.63c 168.7±1.75cd 173.5±1.73d
D 152.2±1.1a 157.53±1.15b 162.54±1.17c 167.55±0.86cd 172.64±0.81d
A Serum copper (µg/dl) 61.99±0.88a 61.83±0.82a 61.93±0.83a 61.99±0.84a 62.03±0.84a
B 62.39±1.48a 65.17±1.51ab 68.11±1.46b 71.48±1.16bc 75.38±1.28c
C 62.63±1.5a 66.94±1.2ab 71.95±1.29bc 76.16±1.31c 80.61±1.05cd
D 60.88±1.38a 66.86±1.13ab 72.33±1.15bc 77.77±0.85c 83.54±0.8cd
A Serum sodium level (mmol/L) 124.06±1.4a 123.67±1.31a 123.5±1.21a 123.72±1.16a 123.84±1.13a
B 125.93±0.9a 127.79±0.94ab 129.5±0.95b 131.5±0.97bc 133.18±0.8bc
C 123.51±1.18a 125.91±0.98a 128.53±1.01ab 131.09±0.94bc 134.1±1.05c
D 124.01±1.31a 128.83±1.38ab 132.05±1.53bc 135.62±1.58c 139.14±1.52d
A Serum potassium level (mmol/L) 3.86±0.15a 3.84±0.15a 3.85±0.15a 3.86±0.15a 3.87±0.15a
B 3.22±0.17a 4.37±0.15bc 4.5±0.15c 4.58±0.15cd 4.69±0.15d
C 3.59±0.15a 4.01±0.08ab 4.24±0.07b 4.52±0.09cd 4.75±0.07d
D 3.92±0.09a 4.29±0.13b 4.5±0.11cd 4.69±0.09d 4.93±0.09e
A Serum chloride level (mmol/L) 69.79±0.95a 69.68±0.97a 69.77±0.99a 69.87±0.98a 70.02±0.93a
B 71.27±1.89a 74.41±1.92b 76.75±2.01bc 80.27±2.24c 84.92±2.34d
C 71.09±2.09a 74.76±1.94b 79.02±2.07c 84.16±1.51d 88.91±1.12e
D 69.37±1.64a 74.95±1.69b 81±1.78cd 88.67±2.24e 96.55±2.08f

In vivo study of HESEZA (150 and 250 mg/kg b_ wt) on EPG in experimental groups at different days of treatment_

Days of observation
Group 0th 7th 14th 21st 28th
A 585±59.15a 591.67±63.21a 601.67±64a 586.67±59.48a 573.33±45.07a
B 639±33.64a 470±18.26ab 326.33±12.33bc 151.5±13.58c 20.33±3.87d
C 603.33±32.52a 389.67±47.14b 154.83±40.63c 46.5±19.19d 0±0
D 516.67±31.16a 121.33±9.61b 8±1.46c 0±0 0±0

In vitro anthelmintic activity of different concentration of HESEZA against P_ cervi_

Drug/Extract Conc (μg/ml) No of parasite showing motility (in minutes)

0 30 60 90 120 150 180 210 240 270 300
Control - 6 6 6 6 6 6 6 6 6 6 6
Z. armatum (Hydro-ethanolic seed extract) 50 6 6 6 6 6 6 6 6 5 4 3
150 6 6 6 6 6 5 5 4 3 2 1
300 6 6 6 5 4 3 1 0 0 0 0
500 6 5 4 3 1 0 0 0 0 0 0
1000 6 4 3 1 1 0 0 0 0 0 0
3000 6 3 2 1 0 0 0 0 0 0 0
Oxyclozanide 10−5 M 6 2 0 0 0 0 0 0 0 0 0

In vivo study of HESEZA (150 and 250 mg/kg b_ wt) on body weight in experimental groups at different days of treatment_

Days of observation
Group 0th 7th 14th 21st 28th
A 7.77±0.17a 7.78±0.17a 7.81±0.17a 7.82±0.18a 7.84±0.18a
B 7.56±0.25a 8.04±0.21ab 8.61±0.23ab 9.25±0.23b 9.84±0.24b
C 7.93±0.15a 8.45±0.17ab 9.11±0.2b 9.85±0.21b 10.88±0.26bc
D 7.79±0.2a 8.36±0.2ab 9.32±0.3b 10.26±0.19bc 11.37±0.18c

Qualitative phytochemical analysis of HESEZA_

SI. No Active principle Test applied Result
1. Alkaloid Hager's test Positive
2. Terpenoids Salkowski test Positive
3. Diterpenes Copper acetate test Positive
4. Flavonoids Ferric chloride test Positive
5. Steroid Salkowski test Positive
6. Tannins Ferric chloride test Positive
7. Glycosides Sodium hydroxide test Positive
8. Saponins Frothing test Negative
9. Phenolic compound Ferric chloride test Positive

In vitro effect of different concentration of HESEZA on various biochemical parameters following 4 hours of incubation of adult P_ cervi_

Biochemical parameters Control Standard (Oxyclozanide 10−3 M) Concentrations (μg/ml) of seed extract
100 300 1000 3000
Glucose uptake (mg/100mg of tissue) 1.52±0.05a 1.08±0.03e 1.45±0.07a 1.34±0.07b 1.19±0.09c 1.09±0.08d
Glycogen content (mg/100mg of tissue) 2.54±0.03a 1.84±0.09e 2.47±0.04a 2.30±0.08b 2.10±0.03c 1.87±0.07d
Lactic acid production (μg/g) of tissue 80.47±5.18a 116.28±4.34e 85.27±4.87a 97.45±6.40b 108.16±4.18c 116.45±3.27e
Acetylcholinesterase activity (mM of Ach hydrolyzed/g weight/h) 92.09±5.70a 59.17±4.48e 91.00±4.18a 88.55±4.89ab 83.00±4.19ab 70.64±3.66c