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In vitro antigiardial and antirotaviral activity of Psidium guajava L. leaves

Authors:
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VOLUME 43 OCTOBER 2011 ISSUE 5
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616 Indian Journal of Pharmacology | October 2011 | Vol 43 | Issue 5
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In vitro
antigiardial and antirotaviral activity of
Psidium guajava
L. leaves
Sir,
Psidium guajava L., family Myrtaceae, is used widely in
traditional medicine throughout the world for a number of
gastrointestinal ailments.[1] The antidiarrhoeal activity of the
aqueous extract of the leaves has been studied and various
mechanisms of action, namely antimicrobial activity, antispasmodic
activity, inhibition of acetylcholine release, inhibition of bacterial
colonization, and production/action of bacterial enterotoxins
have been proposed.[1,2] The antigiardial and antirotaviral studies
with P. guajava leaves from different geographical locations have
been previously reported.[1] Because of known differences in
the biological efficacy of medicinal plants due to differences in
geographical locations we screened an Indian variety of P. guajava
leaves for its in vitro antigiardial and antirotaviral activity.
Leaves of P. guajava variety Sardar collected from Parinche
valley, Maharashtra, India were used. A hot aqueous extract
(decoction) was used for the assays based on its field use as
revealed during an ethnobotanical survey carried out by us.[3] The
decoction was prepared by boiling 1 g of shade dried leaves in
16 mL double distilled water until the volume became 4 mL.[4]
To replicate field conditions a fresh decoction was prepared
every time. The decoction was centrifuged and filtered through a
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Figure 1: Antigiardial and antirotaviral activity of a decoction of Psidium guajava L. leaves (a) Anti-giardial activity; C: Control, trophozoites in
medium alone; M: Trophozoites incubated with metronidazole (10 g/ml). Values represent mean standard error (n = 3) of percentage viable
trophozoites relative to control (100%); *P < 0.05. (b) Anti-rotaviral activity; C: Control, rotavirus infected MA-104 cells in medium alone. Values
represent mean standard error (n = 3) of percentage dead MA-104 cells relative to control (100%); *P < 0.05.
Letters to the Editor
617Indian Journal of Pharmacology | October 2011 | Vol 43 | Issue 5
membrane of 0.22 m pore size before use. The percent yield with
respect to the starting dried plant material was 10.8% 0.5%
(w/w); the dry weight was (27.0 1.25 mg/mL) (n = 25).
For the biological assays a 0.1% (0.027 0.001 mg/mL), 1%,
(0.270 0.013 mg/ml), 5% (1.350 0.063 mg/mL) and 10%
dilutions (2.7 0.125 mg/mL) prepared in appropriate media
were used for each experiment.
For the antigiardial assay, Giardia lamblia P1 trophozoites
(kindly provided by Dr. P. Das, National Institute for Cholera and
Enteric Diseases, Kolkata, India) were incubated in the absence
(control) and presence of different dilutions of the decoction in
Diamond’s TYI-SS medium. After 24 h, viability was estimated
using trypan blue stain. Three independent experiments were
carried out with duplicate tubes for control and each dilution
of the decoction. Metronidazole (10 g/mL) was used as the
antigiardial control.
The entry and subsequent survival of simian rotavirus SA-11
(kindly provided by Dr. S. Kelkar, National institute of Virology,
Pune, India) in MA-104 cells was assayed by the neutral red
uptake assay.[4] Three independent experiments were carried
out with triplicate wells for control as well as each dilution of
the decoction.
The results are expressed as the mean ± standard error
of the percent values from three independent experiments.
The percentage in each experiment was calculated using the
formula (C or T)/C × 100, where C is the mean value of the
duplicate/triplicate readings of the control group and T is mean
value of the duplicate/triplicate readings of the test (dilutions
of the decoction) groups. Hence, the values of test groups have
been represented as percentages relative to control (100%).
Data wer e analyzed by ANOVA and Dunnett’s post-test using
Prism 4.0 (GraphPad, Inc., San Diego, CA, USA). P 0.05 was
considered as statistically significant.
The decoction affected the viability of G. lamblia trophozoites
at 5% and 10% concentrations [Figure 1a]. The antigiardial
activity though statistically significant, was lesser than that of
metronidazole. Rotavirus, following entry, causes death of MA-104
cells. The decoction decreased cell death in virus infected cells at
5% and 10% concentrations indicating that it had antirotaviral
activity at these concentrations [Figure 1b]. The decoction at these
concentrations was not cytotoxic to MA-104 cells (data not shown).
Results of the study thus showed that the decoction of
P. guajava leaves has antigiardial and antirotaviral activity.
Hence, combining this data with other reported studies,[1,2] it
can be suggested that P. guajava leaves have a broad spectrum
of antimicrobial action and thus could be effective in controlling
diarrhoea due to a wide range of pathogens.
Tannaz J. Birdi, Poonam G. Daswani, S. Brijesh,
Pundarikakshudu Tetali1
The Foundation for Medical Research, Mumbai,
1Naoroji Godrej Centre for Plant Research,
Lawkin Ltd. Campus, Satara, Maharashtra, India
Correspondence to:
Correspondence to:
Dr. Tannaz J Birdi,
E-mail: fmr@fmrindia.org
References
1. Gutiérrez RM, Mitchell S, Solis RV.
Psidium guajava
: A review of its traditional
uses, phytochemistry and pharmacology. J Ethnopharmacol 2008;117:1-27.
2. Birdi T, Daswani P, Brijesh S, Tetali P, Natu A, Antia N. Newer insights into the
mechanism of action of
Psidium guajava
L. leaves in infectious diarrhoea. BMC
Complement Altern Med 2010;10:33.
3. Tetali P, Waghchaure C, Daswani PG, Antia NH, Birdi TJ. Ethnobotanical survey
of antidiarrhoeal plants of Parinche valley, Pune district, Maharashtra, India.
J Ethnopharmacol 2009;123:229-36.
4. Brijesh S, Daswani P, Tetali P, Antia N, Birdi T. Studies on the antidiarrhoeal activity
of
Aegle marmelos
unripe fruit: Validating its traditional usage. BMC Complement
Altern Med 2009;9:47.
DRESSing up to phenytoin
Sir,
The term Drug Related Eosinophilia with Systemic Symptoms
(DRESS) describes a potentially life threatening complication
developing secondary to a drug reaction. We describe a case
of DRESS following phenytoin administration. An 86 year old
female, a known case of hypertension, Chronic kidney disease
(CKD), stroke, and seizure disorder (recently detected) presented
with polymorphic lesions, erythematous macules, widespread
superficial erosions and flaccid pustules on trunk and extremities,
with extensive erosions and hemorrhagic crusts on the lips
[Figure 1] and conjunctival xerosis with erosions on the eyelids
which developed in the last three to four days. Treatment
included phenytoin (100 mg thrice daily for the last three weeks),
aspirin, clopidogrel, amlodipine, prazosin and piracetam. She
also had generalized lymphadenopathy and fever (Temperature
100 degree. F). Her laboratory investigations showed Sodium
bicarbonate - 5.5 mmol/l, Haemoglobin - 7.7 g/dl, Total White
Blood Cell count - 20,200/L (polymorphs 80.6%, lymphocytes
3.4%, and eosinophils 10.4%), aspartate aminotransferase
140 IU/L, alanine aminotransferase – 176 IU/L, alkaline
phosphatase - 340 IU/L, Total Bilirubin - 1.6 mg/dl, direct
bilirubin - 0.98 mg/dl, serum creatinine - 3.4 mg/dl and
urea - 215 mg/dl. Supportive treatment with hydration, steroids
(methylprednisone 1 g per day for three days followed by
hydrocortisone 100 mg three times a day), antibiotics and
hemodialysis was started. She developed severe sepsis,
respiratory failure, cardiac arrest and succumbed to death.
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... [15][16][17] Our previous studies with guava leaves demonstrated that its decoction acts against different diarrhoeal pathogens including Shigella and exhibits a wide spectrum of antidiarrhoeal activity. 18,19 In the present study, a decoction prepared from guava leaves was tested in vitro for its ability to combat antibiotic-resistant clinical isolates of Shigella spp. Towards this, the decoction was studied for its bactericidal action and effect on the bacterial invasion of epithelial cells. ...
... 33,34 Our previous studies also demonstrated the anti-diarrhoeal activity of guava leaves. 18,19,[35][36][37] These included the in vitro effect of GLD on the viability of S. flexneri and a reduction in the invading ability of the bacterium to HEp-2 cells. 18 Based on the promising results with the reference strain used, the current study was undertaken to elucidate the effect of guava leaves against drug-resistant clinical Shigella isolates. ...
Article
Background: Although shigellosis is self-limiting, antibiotics are recommended to minimize the severity of symptoms and reduce mortality rates. However, due to the increasing reports of antibiotic resistance, alternative approaches are needed to combat shigellosis. Interest for research on medicinal plants has increased in recent years, and hence, they can be explored to treat this infectious diarrhoea. Objective: To study the effect of Psidium guajava L. (guava) leaf decoction (GLD) on the antibiotic-resistant clinical isolates of Shigella spp. Materials and Methods: A total of 43 isolated Shigella spp. from diarrhoeal patients were used in this study. The effect of GLD on the bacterial viability was initially assessed. The isolates were divided into two categories: sensitive and resistant to GLD. For sensitive isolates, antibacterial activity of GLD was evaluated while for resistant strains, the ability of GLD for reducing the bacterial invasion of the HEp-2 cell line underwent an investigation. Results: Among the 43 Shigella isolates, GLD affected the growth of 23 strains. The invasion of 9 strains from the 20 remaining resistant isolates was unaffected. Although the number of isolates was less, the data suggested that isolates belonging to S. flexneri serogroup were more sensitive to GLD in comparison with other spp (i.e., sonnei, boydii, and dysenteriae). Conclusion: The results of this study revealed the efficacy of GLD against drug-resistant Shigella spp. and thus could be considered for the treatment of diarrhoea. GLD can be a cost-effective alternative to antibiotics.
... Studies undertaken at the FMR have revealed that guava leaf decoction (GLD) shows anti-rotaviral, anti-giardial [24] and antibacterial activity against V. cholerae and Shigella sp. [25] Although the decoction did not show bactericidal activity against E. coli, it inhibited the colonization and production of labile toxin and also inhibited the IL-8 production [25,26]. ...
... Data from literature as well studies undertaken by FMR have indicated that guava leaves can be used as an alternative to treat infectious diarrhoea of varied etiologies; bacterial, viral and protozoal [24,25,49]. Studies have also documented its use in physiological diarrhoea [50,51]. ...
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Background Diarrhoea is amongst the first ten causes of death and its treatment faces an increased threat of drug resistance. Previous studies on the guava leaf decoction (GLD) revealed its suitability for use in infectious diarrhoea of unknown etiology. Objective The objective of this trial was to establish efficacy, dose and safety of GLD prepared from the Indian Sardar variety in adults with acute infectious diarrhoea. Methods The current trial was an open efficacy randomized 5-day, parallel group multi-arm interventional study. Amongst 137 adults (18–60 years) suffering with acute diarrhoea, 109 were included (57% females, 43% males). Three doses of GLD (6-leaf, 10-leaf and 14-leaf) were compared with controls receiving oral rehydration solution. Decrease in stool frequency and improvement in consistency were the outcomes measured. The data was analyzed using ANOVA, Tukey's post-hoc test, Kruscal-Wallis test and Chi-Square test where applicable. Results The trial showed that the 14-leaf (7.4 g) decoction was the most effective. Administration of the decoction, thrice daily helped the patients regain normalcy in 72 h as opposed to 120 h in controls. Safety of the intervention was reflected by normal levels of haemoglobin, liver and kidney parameters. No adverse events were reported. Conclusion The 14 leaves decoction was a safe treatment for adult acute uncomplicated diarrhoea of unknown etiology. Moreover due to component synergy and divergent mechanisms of action, it could possibly combat the generation of drug resistance and destruction of gut microbiota. Hence GLD has the potential for development as a first line treatment for diarrhoea. Trial registration Trial was registered with Clinical Trials Registry - India (CTRI registration number: CTRI/2016/07/007095). The trial was retrospectively registered.
... Ripened guava contains 2-5 times more vitamin C (150-200 mg/100 g of pulp) than citrus fruits and also good source of phosphorus, iron and calcium (Kaur et al. 2018;Kumari and Choudhary 2019). Different parts of guava tree, especially leaves are also beneficial for curing the diseases like diarrhoea, dysentery and gastroenteritis (Birdi et al. 2011;Singh and Marar 2011). ...
Guava (Psidium guajava L.) wilt is extremely severe and soil borne disease, i.e. difficult to control once the symptoms appear on the plant foliage. It is mainly caused by the fungi like Fusarium oxysporum f. sp. psidii, Fusarium solani, Gliocladium roseum, Cephlosporium sp., Nalanthamala psidii and Gliocladium roseum. Several biocontrol agents based on live bacteria and their enzymes and bioactive compounds released by various fungi reported as the antagonistic strategies against the wilt-causing organisms. Chemical control of guava wilt is also feasible to some extent but the disease may reoccur more aggressively due to the presence of fungal spores in the soil. On the other hand, some well-accepted physical methods, like removal and burning of whole plant are extreme steps to control the disease which cause major financial loss. This review is critically based on the different control measures of guava wilt that have been used in laboratory studies as well as in the guava fields.
... Guava leaves have been used globally in the treatment of gastrointestinal disorders [41,42]. Our work which included a clinical trial has confirmed that guava is a promising anti-diarrhoeal plant exhibiting a wide spectrum of activity [8,[43][44][45][46]. However, its functional components largely remained unknown, mainly due to chemical complexity and possible synergism. ...
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Background Herbal medicines are fast gaining popularity. However, their acceptability by modern practitioners is low which is often due to lack of standardization. Several approaches towards standardization of herbals have been employed. The current study attempted to recognize key peaks from ¹ H NMR spectra which together would comprise of a spectral fingerprint relating to efficacy of Psidium guajava (guava) leaf extract as an antidiarrhoeal when a number of unidentified active principles are involved. Methods Ninety samples of guava leaves were collected from three locations over three seasons. Hydroalcoholic (water and ethanol, 50:50) extracts of these samples were prepared and their ¹ H NMR spectra were acquired. Spectra were also obtained for quercetin, ferulic acid and gallic acid as standards. Eight bioassays reflecting different stages of diarrhoeal pathogenesis were undertaken and based on pre-decided cut-offs, the extracts were classified as ‘good’ or ‘poor’ extracts. The bioactivity data was then correlated with the ¹ H NMR profiles using Regression or Orthogonal Partial Least Square-Discriminant Analysis (OPLS-DA). Results OPLS-DA showed seasonal and regional segregation of extracts. Significant models were established for seven bioassays, namely those for anti-bacterial activity against Shigella flexneri and Vibrio cholerae , adherence of E. coli , invasion of E. coli and S. flexneri and production and binding of toxin produced by V. cholerae . It was observed that none of the extracts were good or bad across all the bioassays. The spectral analysis showed multiple peaks correlating with a particular activity. Based on NMR and LC-MS/MS, it was noted that the extracts contained quercetin, ferulic acid and gallic acid. However, they did not correlate with the peaks that segregated extracts with good and poor activity. Conclusions The current study identified key peaks in ¹ H NMR spectra contributing to the anti-diarrhoeal activity of guava leaf extracts. The approach of using spectral fingerprinting employed in the present study can thus be used as a prototype towards standardization of plant extracts with respect to efficacy.
... Hence, alternative safe therapies for rotavirus infections have become the subject of ongoing research (Alfajaro et al., 2014). Pinus koraiensis, Lomatium dissectum, Artocarpus integrifolia, Myristica fragrans, Spongias lutea, Tylosema esculentum, Byrsonima verbascifolia, Myracrodruon urundeuva Allemão, Eugenia dysenterica, Hymenaea courbaril, and Achillea kellalensis were reported to hinders rotaviral strains (Cecílio et al. 2012;Chingwaru et al. 2011;Gandhi et al., 2016;Goncalves et al., 2005;Taherkhani et (Birdi et al., 2011;Brijesh et al., 2009;Civra et al., 2017;El-Baz et al., 2015;Knipping et al., 2012;Mohamed et al., 2015;Pilau et al., 2011;Roner et al., 2010;Téllez, Téllez, Vélez, & Ulloa, 2015). Besides, Vaccinium macrocarpon, Vitis labrusca, and Myracrodruon urundeuva (Cecílio et al., 2016;Lipson et al. 2007Lipson et al. , 2011Lipson et al. , 2012 fever, malaise, rash, and common colds. ...
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The pandemic of viral diseases like novel coronavirus (2019-nCoV) prompted the scientific world to examine antiviral bioactive compounds rather than nucleic acid analogous , protease inhibitors, or other toxic synthetic molecules. The emerging viral infections significantly associated with 2019-nCoV have challenged humanity's survival. Further, there is a constant emergence of new resistant viral strains that demand novel antiviral agents with fewer side effects and cell toxicity. Despite significant progress made in immunization and regenerative medicine, numerous viruses still lack prophylactic vaccines and specific antiviral treatments that are so often influenced by the generation of viral escape mutants. Of importance, medicinal herbs offer a wide variety of therapeutic antiviral chemotypes that can inhibit viral replica-tion by preventing viral adsorption, adhering to cell receptors, inhibiting virus penetration in the host cell, and competing for pathways of activation of intracellular signals. The present review will comprehensively summarize the promising antiviral activities of medicinal plants and their bioactive molecules. Furthermore, it will elucidate their mechanism of action and possible implications in the treatment/prevention of viral diseases even when their mechanism of action is not fully understood, which could serve as the base for the future development of novel or complementary ant-iviral treatments. K E Y W O R D S 2019-nCoV, antiviral chemotypes, medicinal plants, phytomolecules, viral infections
... On calculating the trophozoite count in the treated groups, following the end of therapy, the intestinal parasite density was, noticeably, significantly reduced in GIV that received PGLE. Birdi et al., 2011 proved the in-vitro potential anti-giardia effect of PGL decoction. In the same context, Neiva et al., 2014 concluded that the hydroethanolic PGLE showed moderate in-vitro antigiardia activity at a concentration of 500 lg/mL. ...
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... These animals had inclined levels of HDL-c. Birdi et al., [2,3] recorded that leaves of guava have a broad range of antimicrobial accomplishment that able to prevent diarrhoea. The antimicrobial action of leaves can be accompanying to the presence of flavonoids [10]. ...
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... Birdi et al. [176], reported that the decoction of P. guajava leaves possess anti-rotaviral activity. Since in presence of the extract there was decrease in cell death in infected cultures it was concluded that entry and subsequent survival of simian rotavirus was prevented by the decoction of guava leaves. ...
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Psidium guajava L., Myrtaceae, is used widely in traditional medicine for the treatment of diarrhoea, dysentery, gastroenteritis, stomachaches, and indigestion. However, the effect of the leaf extract of P. guajava on the pathogenesis of infectious diarrhoea has not been studied. The present study evaluates the effect of a hot aqueous extract (decoction) of dried leaves of P. guajava on parameters associated with pathogenicity of infectious diarrhoea. The aim was to understand its possible mechanism(s) of action in controlling infectious diarrhoea and compare it with quercetin, one of the most reported active constituents of P. guajava with antidiarrhoeal activity. The crude decoction and quercetin were studied for their antibacterial activity and effect on virulence features of common diarrhoeal pathogens viz. colonization of epithelial cells and production and action of enterotoxins. Colonization as measured by adherence of enteropathogenic Escherichia coli (EPEC) and invasion of enteroinvasive E. coli (EIEC) and Shigella flexneri was assessed using HEp-2 cell line. The production of E. coli heat labile toxin (LT) and cholera toxin (CT) and their binding to ganglioside monosialic acid (GM1) were studied by GM1-ELISA whereas the production and action of E. coli heat stable toxin (ST) was assessed by suckling mouse assay. The decoction of P. guajava showed antibacterial activity towards S. flexneri and Vibrio cholerae. It decreased production of both LT and CT and their binding to GM1. However, it had no effect on production and action of ST. The decoction also inhibited the adherence of EPEC and invasion by both EIEC and S. flexneri to HEp-2 cells. Quercetin, on the other hand, had no antibacterial activity at the concentrations used nor did it affect any of the enterotoxins. Although it did not affect adherence of EPEC, it inhibited the invasion of both EIEC and S. flexneri to HEp-2 cells. Collectively, the results indicate that the decoction of P. guajava leaves is an effective antidiarrhoeal agent and that the entire spectrum of its antidiarrhoeal activity is not due to quercetin alone.
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Aegle marmelos (L.) Correa has been widely used in indigenous systems of Indian medicine due to its various medicinal properties. However, despite its traditional usage as an anti-diarrhoeal there is limited information regarding its mode of action in infectious forms of diarrhoea. Hence, we evaluated the hot aqueous extract (decoction) of dried unripe fruit pulp of A. marmelos for its antimicrobial activity and effect on various aspects of pathogenicity of infectious diarrhoea. The decoction was assessed for its antibacterial, antigiardial and antirotaviral activities. The effect of the decoction on adherence of enteropathogenic Escherichia coli and invasion of enteroinvasive E. coli and Shigella flexneri to HEp-2 cells were assessed as a measure of its effect on colonization. The effect of the decoction on production of E. coli heat labile toxin (LT) and cholera toxin (CT) and their binding to ganglioside monosialic acid receptor (GM1) were assessed by GM1-enzyme linked immuno sorbent assay whereas its effect on production and action of E. coli heat stable toxin (ST) was assessed by suckling mouse assay. The decoction showed cidal activity against Giardia and rotavirus whereas viability of none of the six bacterial strains tested was affected. It significantly reduced bacterial adherence to and invasion of HEp-2 cells. The extract also affected production of CT and binding of both LT and CT to GM1. However, it had no effect on ST. The decoction of the unripe fruit pulp of A. marmelos, despite having limited antimicrobial activity, affected the bacterial colonization to gut epithelium and production and action of certain enterotoxins. These observations suggest the varied possible modes of action of A. marmelos in infectious forms of diarrhoea thereby validating its mention in the ancient Indian texts and continued use by local communities for the treatment of diarrhoeal diseases.
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