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Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (5): 538-545.doi: 10.3736/jcim20120509

• Original Experimental Research • Previous Articles     Next Articles

In vitro and in vivo evaluation of the inhibitory effect of Limnophila indica (Linn.) Druce on shigellosis

Sandhya Subhadra(), Gouthami Gade, Vinod Kombath Ravindran, Vidya Sravanthi Emani, Saikumar Parre, David Banji#br#   

  1. Department of Pharmacognosy, Nalanda College of Pharmacy, Cherlapally, Nalgonda 508001, Andhra Pradesh, IndiaDepartment of Pharmacognosy, Nalanda College of Pharmacy, Cherlapally, Nalgonda 508001, Andhra Pradesh, India
  • Received:2011-11-11 Accepted:2012-01-06 Online:2012-05-20 Published:2018-06-15
  • Contact: Subhadra Sandhya

Objective: The research aimed to evaluate the anti-Shigella and antacid activities of the methanolic extract of Limnophila indica.

Methods: The whole plant of L. indica was extracted using methanol and then subjected to preliminary chemical screening. The in vitro antibacterial screening on two Gram-positive and two Gram-negative bacteria as well as threeShigella species of which two bacteria were antibiotic-resistant were evaluated by disc diffusion method. Castor oil-induced diarrhoea on Wistar albino rats was performed by using loperamide as a standard control. The in vitroantacid activity was tested by artificial stomach model. The neutralization efficiency, capacity, volume and hydrogen ions consumed were also evaluated.

Results: The preliminary chemical screening on methanolic extract of L.indica showed the presence of phenolic compounds, flavonoids, alkaloids, fats and oils. It was proved to be a potent antibacterial agent against the four bacterial strains. Screening against Shigella species revealed that it was a powerful antibacterial agent towards antibiotic-resistant Shigella species. In the case of in vivo antidiarrheal activity, the plant has shown a dose-dependent activity and the lowest dose at 100 mg/kg gave a much better result than loperamide (P<0.01). The in vitro antacid study showed a mild activity.

Conclusion: As the plant L. indica has been proved to be a competent antibacterial as well as a compelling antidiarrheal agent with mild antacid activity, this plant can be very well suggested to be an eminent substitute for the various synthetic anti-dysentery and antidiarrheal agents available in the market.

Key words: Limnophila, Shigella dysenteriae, antidiarrheals, antacids, dyspepsia, rats


Group n Zone of inhibition (mm)
Bacillus subtilis Staphylococcus
Escherichia coli
Normal control 3 0.54±1.00 1.20±2.00 1.50±1.53 0.85±0.58
Gentamycin (1 mg/mL) 3 15.67±1.16** 14.67±0.58** 14.67±0.58** 15.00±0.00**
Limnophila indica methonal extract (25 mg/mL) 3 1.67±1.16** 11.67±1.53** 11.67±1.53** 9.33±1.53**
Limnophila indica methonal extract (50 mg/mL) 3 7.33±1.16** 12.00±0.00** 11.67±0.58** 11.67±0.58**
Limnophila indica mehonal extract (75 mg/mL ) 3 10.67±1.16** 13.00±1.00** 12.67±1.16** 14.33±0.58**
Limnophila indica methonal extract (100 mg/mL) 3 12.00±2.00** 15.00±1.00** 14.67±1.16** 20.00±1.00**


Group n Zone of inhibition (mm)
Shigella flexneri Shigella dysentery Shigella boydii
Normal control 3 0.68±0.58 1.00±0.58 0.62±1.00
Ceftazidime (1 mg/mL) 3 15.00±0.00** 28.00±1.00** 20.00±0.00**
Limnophila indica methonal extract (25 mg/mL) 3 11.33±0.58** 11.00±0.00** 10.33±0.58**
Limnophila indica methonal extract (50 mg/mL) 3 12.33±0.58** 12.33±0.58** 12.33±0.58**
Limnophila indica mehonal extract (75 mg/mL) 3 14.33±0.58** 14.66±0.58** 14.33±0.58**
Limnophila indica methonal extract (100 mg/mL) 3 18.33±0.58** 18.67±0.58** 19.00±0.00**


Minimum inhibitory concentration (μg/mL) Grade
≤20 More active
20 to 100 Active
100 to 250 Moderately active
250 to 500 Weakly active
>500 Inactive


Microorganism MIC (μg/mL)
Bacillus subtilis 335
Staphylococcus aureus 180
Pseudomonas aerugenosa 190
Escherichia coli 225
Shigella flexneri 230
Shigella dysentery 233
Shigella boydii 235


Group n Total number of defecation Number of wet defecation
Control (1 mL/kg saline) 6 11.40±0.24 8.00±0.32
Standard (3 mg/kg loperamide) 6 2.20±0.58△△ 0.60±0.24△△
Limnophila indica methanol extract (100 mg/kg) 6 1.60±0.40△△ 0.20±0.20△△
Limnophila indica methanol extract (200 mg/kg) 6 0.80±0.37△△ 0

Figure 1

Neutralization effect of methanol extract of Linmophila indica on artificial gastric juice C: Control group; S1: 100 mg/kg sodium bicarbonate group; S2: 200 mg/kg sodium bicarbonate group; E1: 100 mg/kg Limnophila indica methanol extract group; E2: 200 mg/kg Limnophila indica methanol extract group. Data are expressed as mean±standard deviation; n=6; ▲▲P<0.01, vs control group."

Figure 2

Duration of neutralizing effect tested by modified Vatier‘s artificial stomach model C: Control group; S1: 100 mg/kg sodium bicarbonate group; S2: 200 mg/kg sodium bicarbonate group; E1: 100 mg/kg Limnophila indica methanol extract group; E2: 200 mg/kg Limnophila indica methanol extract group. Data are expressed as mean±standard deviation; n=6; ▲▲P<0.01, vs control group."

Figure 3

Volume of artificial gastric juice consumed by methanol extract of Limnophila indica (Fordtran’s in vitro titration model) C: Control group; S1: 100 mg/kg sodium bicarbonate group; S2: 200 mg/kg sodium bicarbonate group; E1: 100 mg/kg Limnophila indica methanol extract group; E2: 200 mg/kg Limnophila indica methanol extract group. Data are expressed as mean±standard deviation; n=6; ▲▲P<0.01, vs control group; □□P<0.01, vs S1 group."

Figure 4

Amount of hydrogen ions consumed by methanol extract of Limnophila indica C: Control group; S1: 100 mg/kg sodium bicarbonate group; S2: 200 mg/kg sodium bicarbonate group; E1: 100 mg/kg Limnophila indica methanol extract group; E2: 200 mg/kg Limnophila indica methanol extract group. Data are expressed as mean±standard deviation; n=6; ▲▲P<0.01, vs control group."

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