China rose. Hibiscus rosa-sinensis. Leaf. 18. ... fermentation, sucrose fermentation, Voges-Proskauer (VP) test, methyl red (MR) reaction, hydrogen sulfide (H2S) ...
Advances in Bioscience and Bioengineering ISSN 2201-8336 Volume 1, Number 1, 2013, 1-24
Antimicrobial Activity of Some Medicinal Plants against Multi Drug Resistant Human Pathogens Md. Al Nayem Chowdhury, M. Ashrafuzzaman, Md. Hazrat Ali, Lutfun Nahar Liza, Kazi Mohammad Ali Zinnah Department of Genetic Engineering and Biotechnology, School of Life Sciences, Shahjalal University of Scienceand Technology, Sylhet-3114, Bangladesh
Corresponding author: M. Ashrafuzzam an, Departm ent of Genetic Engineering and Biotechnology, School of Life Sciences, Shahjalal University of Scienceand Technology, Sylhet-3114, Bangladesh
Abstract The present study was conducted with a view to evaluate the therapeutic potentials of twenty six plant extracts traditionally used in Bangladesh against human pathogenic bacteria Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Proteus mirabilis by disc diffusion method. Out of twenty six plant extracts eight crude plant extracts namely Allamanda cathartica (leaf), Allium sativum (bulb), Citrus limon (fruit), Tam arindus indica (fruit), Prunus domestica (Fruit), Averrhoa carambo la (fruit), Piper betle (leaf) and Terminalia arjuna (leaf) were found to exhibit potential antimicrobial properties against the isolated human clinical bacterial isolates whereas twelve plant extracts were failed to show any antibacterial activity against any of the isolates of Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa followed by ten plant species in case of Proteus mirabilis. The maximum antimicrobial activity was found up to 80% in Tam arindus indica in case of Klebsiella pneumoniae, Pseudomonas aeruginosa and Proteus mirabilis isolates meanwhile it is Averrhoa carambola which exhibited maximum activity against Escherichia coli isolates. Among the plant species tested, Tam arindus indica showed the most promising result. These results dem onstrate the antimicrobial potential of the plants and hence lend support for the use of them in traditional m edicine. Sensitivity of the bacterial isolates was also evaluated for eight comm ercial antibiotic discs where most of the isolates found to develop resistance against multiple comm ercial antibiotics. Among the antibiotics used, ciprofloxacin was the most effective one.
Key word s: Antimicrobial activity, plant extract, human pathogens, multi -drug resistant, Disc diffusion method.
© Copyright 2013 the authors.
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Introduction Diseases are the major cause of death in the developing countries and accounts to 50% of it. Antimicrobial agents are essentially important in reducing the global burden of infectious diseases. However, as resistant pathogens develop and spread, the effectiveness of the antibiotics is diminished. This type of bacterial resistance to the antimicrobial agents poses a very serious threat to public health, and for all kinds of antibiotics, including the major last-resort drugs, the frequencies of resistance are increasing worldwide (Levy et al., 2004; Mandal et al., 2009). Bacterial resistance to antibiotics increa ses mortality likelihood of hospitalization and length of stay in the hospital (Winstanley, 1997). In recent years, drug resistance to human pathogenic bacteria has been commonly reported from all over the world (Piddock and Wise, 1989; Singh et al., 1992; Mulligen et al., 1993; Davis, 1994; Robin et al., 1998). However, the situation is alarming in developing as well as developed countries due to indiscriminate use of antibiotics. Therefore, alternative antimicrobial strategies are urgently needed, and thus this situation has led to a re-evaluation of the therapeutic use of ancient remedies, such as plants (Mandal et al., 2010; Basualdo et al., 2007). Being new, such compounds may not have the problem of microbial resistance. Plant based antimicrobials represent a vast untapped source. The use of plant extract for medicinal treatment has become popular when people realized that the effective life span of antibiotic is limited and over prescription and misuse of traditional antibiotics are causing microbial resistance (Alam et al., 2009). Traditionally used medicinal plants produce a variety of compounds of known therapeutic properties (Iyengar, 1985; Chopra et al., 1992; Harborne and Baxter, 1995). In recent years, antimicrobial properties of medicinal plant s are being increasingly reported from different parts of the world (Grosvenor et al., 1995; Ratnakar and Murthy, 1995; Silva et al., 1996; David, 1997; Saxena, 1997; Nimri et al., 1999; Saxena and Sharma, 1999).At present, nearly 30% or more of the modern
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pharmacological drugs are derived directly or indirectly from plants and their extracts dominate in homeopathic or ayurvedic medicines (Murugesan et al., 2011; Jabeen et al., 2007; Banso, 2009; Ahamunthunisa and Hopper, 2010).Considering the vast potentiality of plants as sources for antimicrobial drugs, this study aimed to detect the antibacterial activities of some natural plant extracts and investigate the effect of some commercial antibiotics against multi-drug resistant human clinical bacterial isolates.
Materials and Methods Collection of bacterial isolates Human clinical bacterial isolates were collected from Popular Diagnostic Center, Sylhet, Bangladesh and aseptically transferred to the USDA (United States Department of Agriculture) Project Laboratory of the Department of Genetic Engineering and Biotechnology, Shahjalal University of Science and Technology, Sylhet for further studies. A total of 40 clinical isolates of presumptive Klebsiella spp. Pseudomonas spp. Proteus spp. and Escherichia coli were collected during February 2012. After collection of all the isolates, were labeled, sub cultured and stored at -200c for further use. Collection of plant materials Plant materials (leaf, bark fruit, rhizome etc.) were collected from different parts of Sylhet district, during February 2012. Plants were authenticated by a taxonomist. Preparation of plant extracts A total of 26 plant extracts were used in this study to screen their antibacterial activity by disc diffusion method (Table 1). The fresh parts of plants such as young leaves, bark, bulb, root, flower, rhizome or petiole (Figure 1) were collected and washed several times with distilled water. The plant parts were cut into small
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pieces and paste was made by using mortar-pestle. The stored leaf extracts were impregnated on filter paper and air dried under laminar air flow cabinet for overnight. Dried filter papers were punched by a punching machine into a size of 6mm diameter. These filter paper discs were used as herbal discs. Table 1: List of medicinal plants used to evaluate antimicrobial activity: Sl. No.
Local name
English Name
Botanical name
Parts of plant used
01.
Allamanda
Allamanda
Allamanda cathartica
Leaf
02.
Arjun
Arjuna
Terminalia arjuna
Leaf and bark
03.
Lobongo
Clove
Syzygium aromaticum
Fruit
04.
Lebu
Lemon
Citrus limon
Fruit and leaf
05.
Daruchini
Cinnamon
Cinnamomus Zeylanicum
Bark
06
Kalojam
Jambul
Syzygium cumini
Fruit
07.
Kalijira
Black cumin
Nigella sativa
Seed
08.
Rosun
Garlic
Allium sativum
Bulb
09.
Peyaj
Onion
Allium cepa
Bulb
10.
Paan
Betel leaf
Piper betle
Leaf
11
Ada
Ginger
Zingiber officinale
Rhizome
12
tejpata
Malabar
Cinnamomum tamala
Leaf
13.
Pepe
Papaya
Carica papaya
Leaf
Chiese gooseberry
Averrhoa carambola
fruit
14.
Kamranga
15.
Thankuni
Asiatic Pennywort
Centella asiatica
Leaf
16.
Tulsi
Tulsi /HolyBasil
Ocimum sanctum
Leaf
17.
Joba
China rose
Hibiscus rosa-sinensis
Leaf
18.
Tetul
Tamarind
Tamarindus indica
fruit
19.
Cha
Tea
Camellia sinensis
Leaf
20.
Rakta chanda
Red sanders
Pterocarpus santalinus
Leaf
21.
Alubokhara
Common Plum
Prunus domestica
Fruit
22.
Stevia
Stevia
Stevia rebaudiana
Leaf
23.
Mahagni
Mahagoni
Swietenia mahagoni
Leaf
24.
Nim
Neem
Azadirachta indica
Leaf
25.
Kop pata
Air plant
Kalanchoe pinnata
Leaf
26.
Kamini
Orange jessamine
Murraya paniculata
Leaf
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Figure 1. Some of the plant parts that were used for herbal sensitivity.
Identification of human clinical bacterial isolates A series of morphological, physiological and biochemical tests were performed to identify the suspected Klebsiella spp. Pseudomonas spp. Proteus spp. and E. coli isolates. The test included Gram staining, motility, oxidase activity, catalase production, oxidation-fermentation
(OF)
test, glucose
fermentation,
lactose
fermentation, sucrose fermentation, Voges-Proskauer (VP) test, methyl red (MR) reaction,
hydrogen sulfide (H2S) production, indole test. All the tests were
conducted according to The Bergey’s Manual of Determinative Bacteriology (Bergey and John, 1994). Antibiogram profiling of human clinical bacterial isolates In vitro susceptibility of the bacterial isolates to various antimicrobial agents was determined by the disc diffusion technique as described by Rahman and Hossain 2010 with slight modification. The nutrient broth culture containing desirable bacterial culture was incubated at 370C for 24-48 hours in a shaking incubator. After obtaining suitable growth of bacterial culture by observation of turbidity, 50μl of broth culture were dropped in the nutrient agar plate. Culture was spread by
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sterile ―L‖ shaped glass rod for the preparation of spread plate culture to set the antibiotic discs. The antibiotic discs were placed on the surface of the medium with sterile forceps and pressed gently to ensure good contact with the surface of the medium. The plates were inverted and placed in an incubator at 37 0C within 15 minutes after the discs were applied. After 16 to 18 hours of incubation, each plate was examined for the determination of the zone of inhibition of the antibiotics. The anti-microbial activities of discs were determined by measuring the zone of inhibition expressed in millimeter. The antibiotic discs used were chloramphenicol, azithromycin, erythromycin, streptomycin, co-trimoxazole, ciprofloxacin, cephradine and gentamicin (Table 2). Table2. Antimicrobial agents and their disc concentrations: Antibacterial agent
Disc concentration (µg) /disc
Chloramphenicol
25
Azithromycin
15
Erythromycin
15
Streptomycin
10
Co-Trimoxazole
25
Ciprofloxacin
5
Cephradine
30
Gentamycin
10
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Antimicrobial activity of plants extract to human clinical bacterial isolates The filter paper discs that were prepared from plant extracts were placed on spread plate culture of test organisms (Klebsiella spp. Pseudomonas spp. Proteus spp. and Escherichia coli) by sterilized forceps. The plates were then allowed to incubate at 37°C for overnight. After 12-24 h of incubation, the plant extract was noted for zone of inhibition for each Klebsiella spp. Pseudomonas spp. Proteus spp. and E. coli isolates. The diameter of the zone of inhibition was measured by measuring scale in millimeter (mm).
Results and Discussion Emergence of multi-drug resistance in human and animal pathogenic bacteria as well as undesirable side effects of certain antibiotics has triggered immense interest in the search for new antimicrobial drugs of plant origin. The study was conducted with a view to investigate the antimicrobial properties of some medicinal plant extracts
against
some
human
clinical
bacterial
isolates
(Klebsiella
spp.
Pseudomonas spp. Proteus spp. and Escherichia coli). Identification of the bacterial isolates was carried out using morphological and biochemical properties. Morphological studies were carried out by microscopic observation and studies on growth characteristics in Petri dishes. Visual observation of both morphological and microscopic characteristics using light microscopy and gram staining properties were performed. On the basis of morphological and biochemical characteristics isolates E1-E10, K1-K10, Ps1- Ps10 and P1- P10 were identified as Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Proteus mirabilis respectively (Table 3). In the present study it was found that isolates E1-E10 were motile, gram negative, catalase positive, oxidase negative, MR test positive,indole test positive and
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negative for VP test. The isolates (E1-E10) were fermentative and were able to ferment glucose, sucrose and lactose but did not produce H 2S in TSI (Table 3). Based on all of the above tests the isolates were identified as E. coli. isolates. The results of these biochemical tests were very similar to those observed by other authors who studied some of the serogroups of E.coli (Silva et al., 1980; Farasat al., 2012). During the course of study it was found that isolates K1-K10 were gram negative, fermentative, non-motile, catalase positive, oxidase negative, VP positive and negative for MR, indole production test. The isolates K1 -K10 were able to ferment glucose, sucrose and lactose and were as identified as Klebsiella pneumoniae (Table 3). Mittal et al., 2011 and Sharmeen et al., 2012 also found similar results in their study. Besides these, it was found that isolates Ps1-Ps10 were motile, gram negative, catalase positive, oxidase positive and negative for MR, VP, indole and H 2S production test. All the isolates (Ps1-Ps10) were oxidative and were able to ferment glucose and sucrose but did not ferment lactose (Table 3). Rest of the bacterial isolates (P1-P10) were motile, gram negative, oxidase negative, catalase positive, MR test positive and negative for VP and indole test. The isolates (P1-P10) were able to produce H2S in TSI but did not ferment glucose, lactose and sucrose (Table 3). Based on the biochemical tests the isolates Ps1-Ps10 and P1-P10 were identified as Pseudomonas aeruginosa and Proteus mirabilis respectively.
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Table 3. Biochemical characterization of human clinical bacterial isolates: Tests
Isolate No. ( E1-
Isolate No. (K1-
Isolate No. (Ps1-
Isolate No. (P1-
performed
E10)
K10)
Ps10)
P10)
Gram’s test
-
-
-
-
Motility
+
_
+
+
Glucose
+
+
+
-
+
+
+
-
+
+
-
-
O-F test
F
F
O
ND
Oxidase test
-
-
+
-
Catalase test
+
+
+
+
MR test
+
-
-
+
VP test
-
+
-
-
Indole test
+
-
-
-
H2S production
-
-
-
+
fermentation Sucrose fermentation Lactose fermentation
Legends: (-) = Negative; (+) = Positive; ND = Not done; F = Fermentative; O = Oxidative. H2S= Hydrogen sulfide; MR = Methyl Red; VP = Voges-Proskauer; O-F = OxidativeFermentative.
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The results of antibiotic susceptibility test of the bacterial isolates indicated that high proportion of test organisms were resistant to streptomycin and gentamicin. 70% of the E. coli, Klebsiella pneumonia and Proteus mirabilis isolates were resistant to streptomycin (Figure 2). Almost 50-60% of all the four groups of bacterial isolates were resistant to gentamycin. Most of the human clinical bacterial isolates were sensitive to ciprofloxacin (Figure 2).
All the E. coli isolates showed resistance to at least one of the eight antibiotics used. Of the 10 isolates, 70% were resistant to 2 antibiotics namely streptomycin and azithromycin, 60% to gentamycin, 50% to erythromycin, 40% to chloramphenicol and cephradine and 20% to ciprofloxacin and co-trimoxazole (Figure 2). The Klebshiella pneumoniae isolates showed variable result in their antibiotic sensitivity pattern against eight commercial antibiotic discs tested. The highest rate of resistance was against streptomycin (70%) followed by 50% to chloramphenicol, Erythromycin, cephradine and gentamicin. Almost 30-40% of the Klebshiella pneumoniae
isolates
were
resistant
to
ciprofloxacin,
co-trimoxazole
and
azithromycin (Figure 2). Most of the isolates exhibited resistant to multiple commercial antibiotics and refereed as "Multidrug-resistant organisms" (MDROs). In reviewed study it is available that the frequency of extended spectrum betalactamase (ESBL) producing Klebshiella spp. were found to be resistant against ciprofloxacin (74.8%) cephalosporin (23%) (Podschun et al., 1998; Alipour-fard and Yeasmin, 2010). In addition, all the bacterial isolates belong to Pseudomonas aeruginosa and Proteus mirabilis demonstrated variable resistance pattern against the antibiotics tested. In case of Pseudomonas aeruginosa isolates, the highest rate of resistance was against erythromycin 80% whereas 70% of Proteus mirabilis isolates were resistant to streptomycin (Figure 2). Among the antibiotics used, ciprofloxacin is the most effective one against almost all of the bacterial isolates.
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parcentage of antibiotic resistance
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Chlm 90 80 70 60 50 40 30 20 10 0
Azi Ery Strp Cip Co E
K
Ps
Cep
P
Gen
Name of bacterial isolates
Figure 2: Percentage of Resistance of human clinical bacterial isolates to commercial antibiotics. Legends: E = E. coli; K = Klebshiella pneumoniae; Ps = Pseudomonas aeruginosa; P = Proteus mirabilis; Chlm = Chloramphenicol; Azi = Azithromycin; Ery = Erythromycin; Strp = Streptomycin; Co = Co-trimoxazole; Cip = Ciprofloxacin; Cep = Cephradine; Gen = Gentamicin. The prevalence of resistant bacteria is significant and deserves more consideration. To overcome this constrain now we are taking shelter to our ancestor’s medicinal practice. According to former customs we are taking interest about herbs. Reviewed studies stated that enormous work has done to screen the antibacterial activity of medicinal plants against human pathogen (Jahan et al., 2007; Khan et al., 2007; Rahman et al., 2008; Misra et al., 2009). The antibacterial efficacy of various plant extracts showed varied level of inhibition against the human pathogenic bacteria. Among the 26 samples tested in the present study, 8 crude plants extracts namely Allamanda cathartica
(leaf), Allium sativum (bulb), Citrus limon
(fruit),
Tamarindus indica (fruit), Prunus domestica (Fruit), Averrhoa carambola (fruit),
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Piper betle (leaf) and Terminalia arjuna (leaf) were found to exhibit potential antimicrobial properties against the isolated human clinical bacterial isolates (Table 4, 5, 6 & 7).
Prunus domestica
Nigella sativa Tamarindus indica Allamanda cathartica
Figure 3.Antibacterial activity of plant extract against isolate no. P4.
Allamanda cathartica
Tamarindus indica Prunus domestica
Averrhoa carambola
Figure 4. Antibacterial activity of plant extract against isolate no.K3. Twelve plants extract viz. Terminalia arjuna (bark), Syzygium cumini, Nigella sativa, Cinnamomum tamala, Carica papaya, Centella asiatica, Ocimum sanctum, Hibiscus
rosa-sinensis,
Camellia
sinensis,
Pterocarpus
santalinus,
Stevia
rebaudiana and Kalanchoe pinnata were failed to exhibit any antimicrobial activity against any of the isolates of Escherichia coli, Klebsiella pneumoniae (Table 4, 5 & 6)
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and Pseudomonas aeruginosa followed by ten plant extracts in case of Proteus mirabilis (Table 7). In case of Escherichia coli and Klebsiella pneumonia isolates, the highest zone of inhibition was 21 mm (Table 4 & 5) which was exhibited by Citrus limon (fruit part). Fruit extract of Tamarindus indica exhibited highest activity against Proteus mirabilis (24 mm) isolates (Table 7) as well as against Pseudomonas aeruginosa (19mm) isolates (Table 6). Fruit extract of Averrhoa carambola showed potential antimicrobial activity against 80% of Escherichia coli bacterial isolates whereas Tamarindus indica, Allium sativum and Citrus limon exhibited against 70% of the Escherichia coli isolates. The maximum antimicrobial activity was found up to 80% in Tamarindus indica in case of Klebsiella pneumoniae, Pseudomonas aeruginosa and Proteus mirabilis isolates (Table 5, 6 & 7). In addition, Tamarindus indica, Allamanda cathartica and Allium sativum were able to show antibacterial activity against 70% of the isolates of Pseudomonas aeruginosa and Proteus mirabilis respectively. The results of present investigation clearly indicate that the antibacterial activity vary with the species of the plants and plant material used. Thus, the study ascertains the value of plants used in ayurveda, which could be of considerable interest to the development of new drugs.
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Table 4. Antimicrobial activities of selected plants extract against E. coli isolates. Name of plants
Name of E.coli isolates with their sensitivity pattern (E1-E10) E1
E2
E3
E4
E5
E6
E7
E8
E9
E10
Allamanda cathartica
13
13
-
-
12
14
-
12
-
10
Terminalia arjuna (leaf)
-
8
15
-
12
-
-
-
10
10
Terminalia arjuna
-
-
-
-
-
-
-
-
-
-
Syzygium aromaticum
8
8
8
-
-
-
-
-
-
11
Citrus limon (fruit)
18
21
15
-
-
-
15
21
15
13
Citrus limon (leaf)
-
-
9
-
9
11
-
-
-
9
Cinnamomus
-
-
9
-
-
-
-
-
-
-
Syzygium cumini
-
-
-
-
-
-
-
-
-
-
Nigella sativa
-
-
-
-
-
-
-
-
-
-
Allium sativum
19
14
-
9
11
-
16
-
16
14
Allium cepa
11
9
-
-
-
-
14
-
9
-
Piper betle.
-
-
11
9
-
8
13
11
11
-
Zingiber officinale
9
-
-
-
-
-
-
9
-
-
Cinnamomum tamala
-
-
-
-
-
-
-
-
-
-
(bark)
Zeylanicum
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Carica papaya
-
-
-
-
-
-
-
-
-
-
Averrhoa carambola
13
13
14
-
11
9
17
-
14
17
Centella asiatica
-
-
-
-
-
-
-
-
-
-
Ocimum sanctum
-
-
-
-
-
-
-
-
-
-
Hibiscus rosa-sinensis.
-
-
-
-
-
-
-
-
-
-
Tamarindus indica
12
17
15
-
-
15
12
12
-
15
Camellia sinensis
-
-
-
-
-
-
-
-
-
-
Pterocarpus santalinus
-
-
-
-
-
-
-
-
-
-
Prunus domestica
11
11
-
-
11
-
-
9
-
-
Stevia rebaudiana
-
-
-
-
-
-
-
-
-
-
Swietenia mahagoni
12
-
9
-
9
-
-
10
-
-
Azadirachta indica
-
-
-
9
-
-
-
-
-
-
Kalanchoe pinnata
-
-
-
-
-
-
-
-
-
-
Murraya paniculata
-
12
-
9
-
-
-
-
-
-
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Table 5. Antimicrobial activities of selected plants extract against Klebsiell pneumoniae isolates. Name of plants
Name of Klebsiell pneumoniae isolates with their sensitivity pattern (K1-K10) K1
K2
K3
K4
K5
K6
K7
K8
K9
K10
Allamanda cathartica
12
12
-
-
-
10
-
-
12
10
Terminalia arjuna (leaf)
14
-
9
-
-
-
-
-
-
12
Terminalia arjuna (bark)
8
8
-
-
-
-
-
8
-
-
Syzygium aromaticum
-
-
-
-
-
-
-
-
-
-
Citrus limon (fruit)
17
14
9
21
9
-
-
16
16
-
Citrus limon (leaf)
-
-
-
-
-
-
-
-
-
-
Cinnamomus Zeylanicum
-
-
-
-
-
11
-
-
11
-
Syzygium cumini
-
-
-
-
-
-
-
-
-
-
Nigella sativa
8
8
-
-
-
-
-
10
-
8
Allium sativum
12
12
-
-
13
-
15
10
13
17
Allium cepa
-
11
13
-
-
9
-
9
-
-
Piper betle
-
9
10
-
9
11
8
-
-
9
Zingiber officinale
-
-
9
11
11
-
-
-
-
-
Cinnamomum tamala
-
-
-
-
-
-
-
-
-
-
Carica papaya
-
-
-
-
-
-
-
-
-
-
Averrhoa carambola
-
14
12
-
15
15
-
18
-
11
Centella asiatica
-
-
-
-
-
-
-
-
-
-
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Ocimum sanctum
-
-
-
-
-
-
-
-
-
-
Hibiscus rosa-sinensis
-
-
-
-
-
-
-
-
-
-
Tamarindus indica
-
13
16
9
-
9
16
11
16
13
Camellia sinensis
9
-
-
9
9
-
-
-
-
-
Pterocarpus santalinus
-
-
-
-
-
-
-
-
-
-
Prunus domestica
-
9
12
-
-
-
12
-
14
11
Stevia rebaudiana
-
-
-
-
-
-
-
-
-
-
Swietenia mahagoni
9
-
-
-
-
-
-
-
10
-
Azadirachta indica
-
-
11
-
-
-
-
-
-
-
Kalanchoe pinnata
-
-
-
-
-
-
-
-
-
-
Murraya paniculata
-
-
-
-
-
-
-
-
-
-
Table 6. Antimicrobial activities of selected plants extract against Pseudomonas aeruginosa isolates. Name of plants
Name
of
Pseudomona s
aeruginosa
isolates
with
their
sensitivity pattern (Ps1-ps10) Ps1
Ps2
Ps3
Ps4
Ps5
Ps6
Ps7
Ps8
Ps9
Ps10
Allamanda cathartica
12
9
13
9
8
12
-
12
9
-
Terminalia arjuna (leaf)
12
12
-
-
-
14
-
9
9
-
Terminalia arjun (bark)
-
9
-
-
-
-
9
-
-
9
Syzygium aromaticum
8
8
-
9
-
-
-
-
-
-
Citrus limon (fruit)
12
13
17
-
11
11
-
17
14
-
Citrus limon (leaf)
-
-
-
-
-
-
-
8
-
-
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Cinnamomus Zeylanicum
-
-
-
9
9
-
-
-
8
-
Syzygium cumini
-
-
-
-
-
-
-
-
-
-
Nigella sativa
-
-
-
-
-
-
-
-
-
-
Allium sativum
16
8
8
-
16
12
-
-
-
16
Allium cepa
8
-
-
-
-
-
11
-
-
11
Piper betle
-
9
12
-
12
-
9
13
-
17
Zingiber officinale
-
-
-
-
-
-
9
-
-
8
Cinnamomum tamala
-
-
-
-
-
-
-
-
-
-
Carica papaya
-
-
-
-
-
-
-
-
-
-
Averrhoa carambola
-
14
20
-
15
-
18
15
-
11
Centella asiatica
-
-
-
-
-
-
-
-
-
-
Ocimum sanctum
-
-
-
-
-
-
-
-
-
-
Hibiscus rosa-sinensis
-
-
-
-
-
-
-
-
-
-
Tamarindus indica
19
19
19
11
-
17
14
19
-
19
Camellia sinensis
-
-
-
-
-
-
-
-
-
-
Pterocarpus santalinus
-
-
-
-
-
-
-
-
-
-
Prunus domestica
-
-
17
14
9
-
-
-
-
11
Stevia rebaudiana
-
-
-
-
-
-
-
-
-
-
Swietenia mahagoni
-
9
-
-
-
8
8
-
-
-
Azadirachta indica
-
-
-
-
-
-
-
-
-
-
Kalanchoe pinnata
-
-
-
-
-
-
-
-
-
-
Murraya paniculata
-
-
11
-
-
-
-
11
-
8
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Advances in Bioscience and Bioengineering
Table 7. Antimicrobial activities of selected plants extract against Proteus mirabilis isolates. Name of plants
Name of Proteus mirabilis isolates with their sensitivity pattern (P1-P10) P1
P2
P3
P4
P5
P6
P7
P8
P9
P10
Allamanda cathartica
14
17
-
9
-
-
-
-
9
11
Terminalia arjuna (leaf)
-
12
12
9
9
-
-
12
-
-
Terminalia arjuna (bark)
-
8
-
-
11
-
-
-
-
-
Syzygium aromaticum
-
-
-
-
-
9
9
-
-
-
Citrus limon (fruit)
-
15
-
17
14
-
16
16
18
-
Citrus limon (leaf)
-
-
-
9
-
-
-
9
-
-
Cinnamomus Zeylanicum
9
9
-
-
-
-
-
-
11
9
Syzygium cumini
-
-
-
-
-
-
-
-
-
-
Nigella sativa
-
-
-
-
-
-
-
-
-
-
Allium sativum
14
10
10
-
10
13
-
16
11
13
Allium cepa
9
-
-
-
-
-
-
9
-
-
Piper betle
-
8
8
-
13
-
11
11
-
11
Zingiber officinale
-
-
9
-
-
-
-
8
8
-
Cinnamomum tamala
-
-
-
-
-
-
-
-
-
-
Carica papaya
-
-
-
-
-
-
-
-
-
-
Advances in Bioscience and Bioengineering
20
Averrhoa carambola
-
9
15
-
12
12
14
9
18
-
Centella asiatica
-
-
-
-
-
-
-
-
-
-
Ocimum sanctum
-
-
-
-
-
-
-
-
-
-
Hibiscus rosa-sinensis
-
-
-
-
-
-
-
-
-
-
Tamarindus indica
15
21
-
24
13
13
21
-
24
17
Camellia sinensis
-
-
-
8
-
-
-
-
-
-
Pterocarpus santalinus
-
-
-
-
-
-
-
-
-
-
Prunus domestica
-
9
11
17
-
9
-
-
-
14
Stevia rebaudiana
-
-
-
-
-
-
-
-
-
-
Swietenia mahagoni
-
-
8
-
8
-
-
11
-
-
Azadirachta indica
-
-
-
-
-
-
-
-
-
-
Kalanchoe pinnata
-
-
-
-
-
-
-
-
-
-
Murraya paniculata
-
8
-
-
-
-
-
8
-
-
Acknowledgements The authors gratefully acknowledged to the authority of USDA Project Laboratory of the department of Genetic Engineering and Biotechnology, Shahjalal University of Science and Technology, Sylhet, Bangladesh for providing facilities to carry out this research work.
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