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In-vitro antibacterial activity screening of herb extracts against foodborne pathogenic bacteria from Thailand.

In recent years, food safety concerns have been focused on pathogens, such as Salmonella which is recognized as a primary cause of food poisoning worldwide and massive outbreaks have been occurred in several parts of the world (1). It was estimated that globally, around 86% of salmonellosis cases are food-borne (2). The S. enterica Typhimurium DT104b has a remarkable ability to survive and becomes the major cause of salmonellosis food-borne illness. This is reflected by the fact that 6.6% of food-borne outbreaks were attributable to this serovar in an international study (3). For S. enterica 4,5,12:i:-(human) US clone is a serotype that appears to be antigenically similar and genetically closely related to S. enterica Typhimurium (which has the antigenic formula 4,5,12:i: 1,2) but lacks expression of the second-phase flagella antigen, which is 1,2 in S. enterica Typhimurium. The S. enterica 4,5,12:i:-(human) US clone was the sixth most common Salmonella serovar among cases of human disease in the United States in 2006 and the fourth most common serovar among human isolates in Spain in 1998. Overall, the prevalence of S. enterica 4,5,12:i:-(human) US clone among human cases has increased considerably in many countries in the world over the last 10 year (4,5,6). While in the last 20 years or so, S. enterica Enteritidis has become the single most common cause of food poisoning. L. monocytogenes is a Gram-positive bacterium, motile by flagella. The 1-10% of population may be intestinal carriers of L. monocytogenes (7). It is quite hardy and resists the deleterious effects of freezing, drying, and heat (7). About 2500 cases of listeriosis occur each year. Most cases of listeriosis and most deaths occur in adults with weakened immune systems, the elderly, pregnant women, and newborns (7). Outbreaks of listeriosis have been linked to a variety of foods especially processed meats and dairy products made from unpasteurized (7). Bacillus cereus is a Gram-positive, may change to Gram-negative when get older, rod-shaped with endospore and aerobe (8). B. cereus themselves not tolerate to physical condition, basic or acidic, but their spores are heat resistant and active in wide range of pH (8). B Growth of Bacillus sp. will gives an enterotoxin. Even B. cereus outbreaks is only 2% of all foodborne illness (8). Escherichia coli bacteria normally live in the intestines of people and animals (9). Most E. coli are harmless and actually are an important part of a healthy human intestinal tract (9). The types of E. coli that can cause diarrhea can be transmitted through contaminated water or food, or through contact with animals or persons (9).

Local herbs become potential natural source of antibiotics and medicinal properties. They have active compound that effective as antimicrobial. This research was aimed to study in vitro the individual antibacterial activity of six Thai local herbs; Tradescantia spathacea (Oyster plant), Andrographis paniculata (Kariyat), Eleocharis acicularis (Needle-Spike Rush), Acacia concinna (Soap pod), Phyllanthus niruri (Stonebreaker), and Tinospora cordifolia (Gulancha), against six foodborne pathogenic bacteria; E. coli ATCC25822, S. enterica Typhimurium U302 (DT1046), S. enterica Enteritidis (human), S. enterica 4,5,12: i-(human) US clone, B.cereus, and L. monocytogenes 10403S under five extraction conditions; 95% ethanol, chloroform, hexane, sterile distilled water, and autoclaving at 121[degrees]C 15 PSI for 15 minutes.

MATERIALS AND METHODS

Plant sample preparation

Herb samples; T. spathacea (Oyster plant), A. paniculata (Kariyat), E. acicularis (Needle-Spike Rush), A. concinna (Soap pod), P niruri (Stonebreaker), and T cordifolia (Gulancha) were collected locally from Pakthongchai, Nakhonratchasima provience, Thailand. The whole part of fresh herb wwas used. Each herb was clean using tab water and cut into small pieces. Then, herbs were stored in refrigerator at 6[degrees]C until use.

Extraction

Each herb was maculated into each solvents; 95% ethanol, chloroform, sterile distilled water, hexane, using ration 1:3 for 48 hours at room temperature and were stirred every 12 hours. For autoclaving, each herb was added into distilled water using ration 1:3 then was autoclaved at 121[degrees]C, 15 PSI for 15 minutes. And then, each herb was filtered and centrifuged at 5000 rpm for 5 minutes. All crude extracts were concentrated using water bath at 45[degrees]C until became slurry. All crude extracts were kept in freezer at -20[degrees]C until use.

Antibacterial Assay

BSAC disc diffusion method for antimicrobial Susceptibility Testing version 8 (10) was used for antibacterial activity assay. The 100 [micro]L of bacteria (approximately 1.5 x [10.sup.8] CFU/ml) was swab on Mueller-Hinton agar (MHA) plate. The 15 [micro]l of 15 mg/ml each crude extracts were used to test antibacterial activity against E. coli ATCC25822, S. enterica Typhimurium U302 (DT1046), S. enterica Enteritidis (human), S. enterica 4,5,12:i-(human) US clone, B.cereus, and L. monocytogenes 10403S.

The 15 [micro]l of each solvents and 100 mg/ml Penicillin-G were used as negative and positive control, respectively. The inhibition zones were measured expressed as cm of inhibition zone to determine the effectiveness of the each crude extracts against each bacterium. The experiment was done in duplicate and three replications independently

Determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC)

The MIC (Minimum Inhibitory Concentration) method was modified from BSAC disc diffusion method for Antimicrobial Susceptibility Testing version 8 (10). Only herb, which showed positive antibacterial activity, was tested for MIC. For MIC test, the herb extracted as following concentration; 0, 32, 64, 128, and 256 [micro]l/ ml, was added into 1 ml fresh NB. Then 100 [micro]l/ml of culture with OD600 reach 0.1 (early log phase) was inoculated, and incubated at 37[degrees]C for 24 hours.

The negative MIC result tubes were chosen for MBC determination by taking 1 loop of negative MIC was streaked on NA then incubated at 37[degrees]C 24 hours. The growth of culture in each plate was observed. The experiment was done in duplicate and three replications independently

RESULTS AND DISCUSSION

The results showed that the antibacterial activity was mostly in 95% ethanol and chloroform extraction conditions in all six crude extracts; T. spathacea, A. paniculata, E. acicularis, A. concinna, P. niruri, and T. cordifolia as in Table 1-6.

T. spathacea or Hawaiian Dwarf, or Rhoeo spathacea is a succulent herb in the Family Commelinaceae T spathacea have been recognized as a functional food particularly in South America u. In Thai folkmedicine, it is used to relieve fever, cough and bronchitis 12. Only 95% ethanol and chloroform T. spathacea crude extract showed antibacterial activity against both positive; B. cereus, and L. monocytogenes 10403S, and negative bacterial; S. enterica Typhimurium U302 (DT1046), S. enterica Enteritidis (human), S. enterica 4,5,12: i-(human) US clone, excepted for E. coli ATCC25822. The highest antibacterial activity was chloroform extract against S. enterica Typhimurium U302 (DT1046) which diameter of clear zone is 10.50 [+ or -] 1.29 mm.

Only 95% ethanol and chloroform T. spathacea crude extract, which had the antibacterial activity, were assayed for minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) as shown in table 2. The MIC of T. spathacea crude extracts was between 32-128 [micro]l/ml against six bacteria while the MBC was between 32-256 [micro]l/ml. T. spathacea extract contained phenolic compounds, tannin and flavonoid (11). It was reported to possess antimicrobial, insecticidal, anti-inflammatory, anticancer and anti-fertility activities (13). T. spathacea have stimulating activity on human lymphocytes (12). The T. spathacea aqueous leaf extracts in the forms of decoction and infusion, were found to have flavonoid and antioxidant activity (11). The decoction and infusion also exhibited antibacterial activity against six species of Gram positive and four species of Gram negative bacteria, notably methicillin-resistant Staphylococcus aureus and Neisseria gonorrhoeae u. The primary anthocyanin in T. spathacea leaves was isolated and identified by NMR to be rhoeonin which contribute as color (14). For T. spathacea, only 95% ethanol and chloroform extracts showed the antibacterial activity against all bacteria, while other three extraction conditions had no antibacterial activity. These reports (11,14) confirm the results in this experiment that photochemical compounds in T. spathacea are the active compound which have antibacterial activity.

A. paniculata or Kariyat or Creat, or Chuanxinlian is a seasonal plant in the Family of Acanthaceae (15). The whole plant has bitter taste. A. paniculata also contained many of flavanoids and polyphenol as their active compounds (15). In this experiment, only 95% ethanol and chloroform A. paniculata crude extracts showed the antibacterial activity against both gram positive and negative bacteria, while other three extraction conditions had the antibacterial activity. The highest antibacterial activity was chloroform extract against S. enterica 4,5,12:i-(human) US clone which diameter of clear zone is 10.75 [+ or -] 0.96 mm as showed in table 3.

Only 95% ethanol and chloroform A. paniculata crude extract, which had the antibacterial activity, were assayed for minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) as shown in table 4. The MIC of A. paniculata crude extracts was between 32-128 [micro]l/ml against six bacteria while the MBC was between 32-256 [micro]l/ml. The major active compound of A. paniculata are lactone group; andrographolide, deoxy-andrographolide, neoandrographolide, dehydroandrographolide (16). Diterpenoids, flavanoids and polyphenols are also major active compound in A. Paniculata (15). The other study reported that the water extract of boiling roots have effective against S. aureus (17). For methanol extract of stem have antibacterial activity against Proteus vulgaris, stem and leaves powder extract have antibacterial activity against Shigella bacteria but it is not effective against Cholera (17). The ethanol extract of leaves have antibacterial activity against S. aureus and E. coli (17). The study of ethanol extract of A. paniculata powder extracted is effective against B. cereus and L. monocytogenes under both normal and osmotic stress (18,19). However, it may be that individual antibacterial flavonoids have multiple cellular targets, rather than one specific site of action; inhibition of nucleic acid synthesis, cytoplasmic membrane function, and energy metabolism (20). There was a study indicated that 2',4'-or 2',6' dihydroxylation of the B ring and 5,7-dihydroxylation of the A ring in the flavanone structure was important for anti-methicillin-resistant S. aureus activity (21). The former study investigated that no antibacterial activity of andrographolide against any of E. coli, Shigella sonnei, S. aureus, P. aeroginosa, S. pneumoniae, S. pyogenes, Legionella pneumophila, and Bordetella pertusis (22). These reports (15,16,17,18,19,20,21) confirm the results in this experiment that photochemical compounds in A. paniculata are the active compound which have antibacterial activity.

E. acicularis or needle spikerush or dwarf hairgrass is in family Cyperaceae. In this experiment, only 95% ethanol and chloroform E. acicularis crude extracts showed the antibacterial activity against both gram positive and negative bacteria, while other three extraction conditions had the antibacterial activity. The highest antibacterial activity was chloroform extract against S. enterica Typhimurium U302 (DT1046) which diameter of clear zone is 10.50 [+ or -] 1.91 mm as showed in table 5. E. acicularis crude extract showed antimicrobial activity against S. aureus with inhibition zone 10 [+ or -] 0.7 mm. and also had antioxidant activity (DPPH-IC50) more than 20 ig/mL (23). E. acicularis's seed crude extract also had antioxidant activity (24).

Only 95% ethanol and chloroform E. acicularis' crude extract, which had the antibacterial activity, were assayed for minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) as shown in table 6. The MIC of E. acicularis' crude extracts was between 32-256 [micro]l/ml against six bacteria while the MBC was between 32-256 [micro]l/ml. Eleocharis sp. contained "a high concentration of a-sitosterol and lupeol but the most dominant component was a-sitostanol (24-ethylcholestan-3a-ol) (25). This plant have no report on antibacterial activity. It means that a-sitosterol and lupeol have antibacterial capacity to against most of bacteria that used in this experiment.

A. concinna or som-poi is important medicinal plant belonging to family Acaciaceae (26). The bark contains high levels of saponins, which are foaming agents that are found in several other plant species (27). In this experiment, A. concinna's leaf was used. It was found out that only 95% ethanol and chloroform A. concinna crude extracts showed the antibacterial activity against both gram positive and negative bacteria, while other three extraction conditions had the antibacterial activity. The highest antibacterial activity was 95% ethanol extract against S. enterica 4,5,12: i-(human) US clone which diameter of clear zone is 10.50 [+ or -] 1.29 mm as showed in table 7.

Only 95% ethanol and chloroform A. concinna crude extract, which had the antibacterial activity, were assayed for minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) as shown in table 8. The MIC of E. acicularis' crude extracts was between 32-256 [micro]l/ml against six bacteria while the MBC was between 32-256 [micro]l/ml. In previous study, the crude extracted of A. concinna pod can inhibit B. subtilis, E.coli, S. aureus, P aeruginosa, and K. pneumonia (28). The other study of bark extract is effective against S. typhi, P. nirabilis, S. aureus, Yersinia, S. epidermis, K. pnemonia, P. aerogenosa, E. coli, and B. subtilis under methanol, ethanol, and chloroform extraction condition. These extraction can extracted active compounds included Phenol, Tannin, fat and fixed oil, Flavanoids, Saponin, and quinine (29). The benzene, methanol and aqueous extracts of fresh pods of A. concinna showed maximum activity against K. pneumoniae, B. subtilis, E. coli, followed by P. aeruginosa and S. aureus (30). Phytochemical study showed that the crude extract of A. concinna pod consisted of alkaloids, flavonoids, saponin and tannin but none of antraquinone and cyanotic glycosides (31). It means that all these compounds have antibacterial activity against most bacteria used in this experiment.

P. niruri or dukong anak or stonebreaker's leaf crude extract only 95% ethanol, chloroform, and sterile distilled water crude extracts showed the antibacterial activity against both gram positive and negative bacteria, while other three extraction conditions had the antibacterial activity. The highest antibacterial activity was chloroform extract against S. enterica Typhimurium U302 (DT1046) which diameter of clear zone is 10.25 [+ or -] 1.50 mm as showed in table 9. The antimicrobial mechanism showed that the bacterial cells, after exposure to the P. niruri extract, showed complete alteration in their morphology, followed by collapse of the cells beyond repair (32). The study revealed that the P. niruri methanolic extract may be an effective antibacterial agent to treat bacterial infections since the extract exhibited significant antimicrobial potency, comparable with that of the standard antibiotic chloramphenicol (32).

95% ethanol, chloroform and sterile distilled water P niruri crude extract, which had the antibacterial activity, were assayed for minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) as shown in table 8. The MIC of P niruri crude extracts was between 32-128 [micro]l/ml against six bacteria while the MBC was between 32-256 [micro]l/ml. P niruri contained flavonoids, alkaloids, terpenoids, lignans, polyphenols, tannins, coumarins and saponins, have been identified from various parts have effect to against the Hepatitis B and other viral infections (33). The other study report that this plant extract is effective against Staphylococcus, Micrococcus, and Pasteurella bacteria under methanol, DCM with methanol (1:1), and aqueous extraction (34). These are some active compounds that can extract from P. niruri, Alkaloids, astragalin, brevifolin, carboxylic acids, corilagin, cymene, ellagic acid, ellagitannins, gallocatechins, geraniin, hypophyllanthin, lignans, lintetralins, and lupeols (34). The P. niruni extracte had antibacterial activity against Candida albicans, B. pumilus, Micrococcus luteus, K. pneumonia, S. aureus, B. subtilis, and E. coli under mathanol extraction condition. These are the active compounds that can extract from this plant flavonoids, alkaloids, terpenoids, lignans, polyphenols, tannins, coumarins and saponins (35). These active compounds had been report as for their antimicrobial activity.

T. cordifolia or guduchi crude extract only 95% ethanol, chloroform, and sterile distilled water crude extracts showed the antibacterial activity against both gram positive and negative bacteria, while other three extraction conditions had the antibacterial activity. The highest antibacterial activity was chloroform extract against S. enterica Typhimurium U302 (DT1046) which diameter of clear zone is 10.50 [+ or -] 2.08 mm as showed in table 11. T. cordifolia composed of difference active compounds, for example, alkaloids, diterpenoid lactones, glycosides, steroids, sesquiterpenoid, aliphatic (36,37). These active compounds had been report as for their antimicrobial activity.

95% ethanol, chloroform and sterile distilled water T. cordifolia crude extract, which had the antibacterial activity, were assayed for minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) as shown in table 8. The MIC of T cordifolia crude extracts was between 32-128 [micro]l/ml against six bacteria while the MBC was between 32-256 [micro]l/ml. T. cordifolia ether extract of stem (aerial part) is effective against Mycobacterium tuberculosis and aqueous extract is effective against E. coli and S. aureus with active compounds included alkaloids, diterpenoid lactones, glycosides, steroids, sesquiterpenoids, phenolics, aliphatic compounds, and polysaccharides (38).

aqueous, ethanol, methanol, and acetone T cordifolia crude extracts had antibacterial activity against E. coli, S. aureus, K. pneumoniae, P. vulgaris, S. typhi, S. flexneri, S. paratyphi, S. typhimurium, P. aeruginosa, E. aerogene, and Serratia marcesensesi with active compounds included a-sitosterol, hydroxy ecdysone, ecdysterone, and giloinsterol (39,40).

CONCLUSION

The six Thai local herbs; T. spathacea, A. paniculata, E. acicularis, A. concinna, P. niruri, and T. cordifolia) showed the antibacterial activity against foodborne pathogenic bacteria. This is the stepping stone for further application in food safety, cosmetic industry, and pharmaceutical industry.

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Patchanee Yasurin * and Sasiwan Piya-Isarakul

Faculty of Biotechnology; Assumption University; Hua-mak, Bangkok, Thailand.

(Received: 10 February 2015; accepted: 20 April 2015)

* To whom all correspondence should be addressed. E-mail:
Table 1. The antibacterial activity (clear zone in mm) of T.
spathacea crude extracts under five different extractions
condition against six foodborne pathogenic bacteria

Microorganism         95% Ethanol           Chloroform        Hexane

E. coli            7.75 [+ or -] 0.96           --              --
  ATCC25822
S. enterica        9.25 [+ or -] 0.50   10.50 [+ or -] 1.29     --
  Typhimurium
  U302 (DT1046)
S. enterica        8.75 [+ or -] 0.96   8.75 [+ or -] 1.71      --
  Enteritidis
  (human)
S. enterica        9.25 [+ or -] 1.50   9.25 [+ or -] 0.96      --
  4,5,12:
  i-(human)
  US clone
B. cereus          8.00 [+ or -] 0.82   8.00 [+ or -] 0.82      --
L. monocytogenes   6.25 [+ or -] 0.50   8.75 [+ or -] 0.96      --
  10403S

Microorganism       Sterile    Autoclave at
                   distilled   121[degrees]C
                     water     15 PSI for 15
                                  minutes

E. coli               --            --
  ATCC25822
S. enterica           --            --
  Typhimurium
  U302 (DT1046)
S. enterica           --            --
  Enteritidis
  (human)
S. enterica           --            --
  4,5,12:
  i-(human)
  US clone
B. cereus             --            --
L. monocytogenes      --            --
  10403S

Table 2. The Minimum Inhibitory Concentration (MIC) and
Minimum Bactericidal Concentration (MBC) of T. spathacea
crude extracts under five different extractions condition
against six foodborne pathogenic bacteria

Microorganism                             MIC ([micro]l/ml)

                                          95% Ethanol   Chloroform

E. coli ATCC25822                             128
S. enterica Typhimurium U302 (DT1046)         128           64
S. enterica Enteritidis (human)               128           32
S. enterica 4,5,12: i-(human) US clone        128           64
B. cereus                                     128           32
L. monocytogenes 10403S                       128           32

Microorganism                             MBC ([micro]l/ml)

                                          95% Ethanol   Chloroform

E. coli ATCC25822                             128
S. enterica Typhimurium U302 (DT1046)         128           64
S. enterica Enteritidis (human)               128           32
S. enterica 4,5,12: i-(human) US clone        128           64
B. cereus                                     256           32
L. monocytogenes 10403S                       128           64

Table 3. The antibacterial activity (clear zone in mm) of A.
paniculata crude extracts under five different extractions
condition against six foodborne pathogenic bacteria

Microorganism             95%               Chloroform        Hexane
                        Ethanol

E. coli            7.25 [+ or -] 0.50           --              --
  ATCC25822
S. enterica        8.25 [+ or -] 0.96   9.50 [+ or -] 1.73      --
  Typhimurium
  U302 (DT1046)
S. enterica        7.25 [+ or -] 0.50   8.50 [+ or -] 1.29      --
  Enteritidis
  (human)
S. enterica        9.25 [+ or -] 1.26   10.75 [+ or -] 0.96     --
  4,5,12:
  i-(human)
  US clone
B. cereus          8.00 [+ or -] 0.82   8.25 [+ or -] 0.50      --
L. monocytogenes   6.50 [+ or -] 0.58   8.25 [+ or -] 2.06      --
  10403S

Microorganism       Sterile     Autoclave at
                   distilled   121[degrees]C
                     water         15 PSI
                               for 15 minutes

E. coli               --             --
  ATCC25822
S. enterica           --             --
  Typhimurium
  U302 (DT1046)
S. enterica           --             --
  Enteritidis
  (human)
S. enterica           --             --
  4,5,12:
  i-(human)
  US clone
B. cereus             --             --
L. monocytogenes      --             --
  10403S

Table 4. The Minimum Inhibitory Concentration (MIC)
and Minimum Bactericidal Concentration (MBC) of A.
paniculata crude extracts under five different
extractions condition against six foodborne
pathogenic bacteria

Microorganism                  MIC ([micro]l/ml)

                           95% Ethanol   Chloroform

E. coli ATCC25822              128           --
S. enterica Typhimurium        128           64
  U302 (DT1046)
S. enterica Enteritidis        128           64
  (human)
S. enterica 4,5,12:            128           32
  i-(human) US clone
B. cereus                      128           32
L. monocytogenes 10403S        128           32

Microorganism                  MBC ([micro]l/ml)

                           95% Ethanol   Chloroform

E. coli ATCC25822              128           --
S. enterica Typhimurium        128           64
  U302 (DT1046)
S. enterica Enteritidis        128           64
  (human)
S. enterica 4,5,12:            128           32
  i-(human) US clone
B. cereus                      256           32
L. monocytogenes 10403S        128           32

Table 5. The antibacterial activity (clear zone in mm)
of E. acicularis crude extracts under five different
extractions condition against six foodborne pathogenic
bacteria

Microorganism                    95%                Chloroform
                               Ethanol

E. coli ATCC25822         8.00 [+ or -] 1.15            --
S. enterica Typhimurium   9.75 [+ or -] 2.22   10.5 0 [+ or -] 1.91
  U302 (DT1046)
S. enterica Enteritidis           --           10.00 [+ or -] 1.63
  (human)
S. enterica 4,5,12:       7.75 [+ or -] 0.96    8.50 [+ or -] 0.58
  i-(human) US clone
B. cereus                 8.50 [+ or -] 0.58    8.00 [+ or -] 0.82
L. monocytogenes 10403S   8.50 [+ or -] 1.00    9.25 [+ or -] 0.96

Microorganism             Hexane    Sterile
                                   distilled
                                     water

E. coli ATCC25822           --        --
S. enterica Typhimurium     --        --
  U302 (DT1046)
S. enterica Enteritidis     --        --
  (human)
S. enterica 4,5,12:         --        --
  i-(human) US clone
B. cereus                   --        --
L. monocytogenes 10403S     --        --

Microorganism             Autoclave at
                          121[degrees]C
                          15 PSI for 15
                             minutes

E. coli ATCC25822              --
S. enterica Typhimurium        --
  U302 (DT1046)
S. enterica Enteritidis        --
  (human)
S. enterica 4,5,12:            --
  i-(human) US clone
B. cereus                      --
L. monocytogenes 10403S        --

Table 6. The Minimum Inhibitory Concentration (MIC)
and Minimum Bactericidal Concentration (MBC) of E.
acicularis crude extracts under five different
extractions condition against six foodborne
pathogenic bacteria

Microorganism                 MIC ([micro]l/ml)

                          95% Ethanol   Chloroform

E. coli ATCC25822             64            --
S. enterica Typhimurium       64            64
  U302 (DT1046)
S. enterica Enteritidis       --            --
  (human)
S. enterica 4,5,12:           128          128
  i-(human) US clone
B. cereus                     128          256
L. monocytogenes 10403S       128          128

Microorganism                 MBC ([micro]l/ml)

                          95% Ethanol   Chloroform

E. coli ATCC25822             64            --
S. enterica Typhimurium       32            32
  U302 (DT1046)
S. enterica Enteritidis       32            32
  (human)
S. enterica 4,5,12:           64            64
  i-(human) US clone
B. cereus                     32           256
L. monocytogenes 10403S       32           256

Table 7. The antibacterial activity (clear zone in mm)
of A. concinna crude extracts under five different
extractions condition against six foodborne pathogenic bacteria

Microorganism         95% Ethanol           Chloroform        Hexane

E. coli           7.25 [+ or -] 0.50    7.25 [+ or -] 0.50      --
  ATCC25822
S. enterica       8.75 [+ or -] 2.06    10.00 [+ or -] 0.82     --
  Typhimurium
  U302 (DT1046)
S. enterica               --            8.25 [+ or -] 0.50      --
  Enteritidis
  (human)
S. enterica       10.50 [+ or -] 1.29   9.25 [+ or -] 0.96      --
  4,5,12:
  i-(human)
  US clone
B. cereus         7.25 [+ or -] 0.50    7.50 [+ or -] 0.58      --
L.                7.25 [+ or -] 0.96    9.25 [+ or -] 0.96      --
  monocytogenes
  10403S

Microorganism      Sterile        Autoclave at
                  distilled   121[degrees]C 15 PSI
                    water        for 15 minutes

E. coli              --                --
  ATCC25822
S. enterica          --                --
  Typhimurium
  U302 (DT1046)
S. enterica          --                --
  Enteritidis
  (human)
S. enterica          --                --
  4,5,12:
  i-(human)
  US clone
B. cereus            --                --
L.                   --                --
  monocytogenes
  10403S

Table 8. The Minimum Inhibitory Concentration (MIC)
and Minimum Bactericidal Concentration (MBC) of A.
concinna crude extracts under five different extractions
condition against six foodborne pathogenic bacteria

Microorganism                             MIC ([micro]l/ml)

                                          95% Ethanol   Chloroform

E. coli ATCC25822                             64            32
S. enterica Typhimurium U302 (DT1046)         256           64
S. enterica Enteritidis (human)               --            --
S. enterica 4,5,12: i-(human) US clone        128           64
B. cereus                                     128           32
L. monocytogenes 10403S                       128           32

Microorganism                             MBC ([micro]l/ml)

                                          95% Ethanol   Chloroform

E. coli ATCC25822                             64            32

S. enterica Typhimurium U302 (DT1046)         256           32
S. enterica Enteritidis (human)               32            32
S. enterica 4,5,12: i-(human) US clone        128           64
B. cereus                                     256          256
L. monocytogenes 10403S                       128          256

Table 9. The antibacterial activity (clear zone in mm)
of P. niruri crude extracts under five different extractions
condition against six foodborne pathogenic bacteria

Microorganism          95% Ethanol           Chloroform        Hexane

E. coli ATCC25822   8.25 [+ or -] 0.50           --              --
S. enterica         7.50 [+ or -] 0.58   10.25 [+ or -] 1.50     --
  Typhimurium
  U302 (DT1046)
S. enterica                 --           9.25 [+ or -] 1.71      --
  Enteritidis
  (human)
S. enterica         8.75 [+ or -] 0.96   9.75 [+ or -] 0.96      --
  4,5,12:
  i-(human)
  US clone
B. cereus           7.50 [+ or -] 0.58   8.25 [+ or -] 0.50      --
L. monocytogenes    8.25 [+ or -] 1.50   10.00 [+ or -] 1.41     --
  10403S

Microorganism            Sterile             Autoclave at
                        distilled        121[degrees]C 15 PSI
                          water             for 15 minutes

E. coli ATCC25822           --                    --
S. enterica         7.75 [+ or -] 0.50            --
  Typhimurium
  U302 (DT1046)
S. enterica                 --                    --
  Enteritidis
  (human)
S. enterica                 --                    --
  4,5,12:
  i-(human)
  US clone
B. cereus                   --                    --
L. monocytogenes            --                    --
  10403S

Table 10. The Minimum Inhibitory Concentration (MIC) and
Minimum Bactericidal Concentration (MBC) of P. niruri crude
extracts under five different extractions condition against
six foodborne pathogenic bacteria

Microorganism               95%          MIC         Sterile
                          Ethanol   ([micro]l/ml)   distilled
                                     Chloroform       water

E. coli ATCC25822           64           --
S. enterica Typhimurium     64           64            256
  U302 (DT1046)
S. enterica Enteritidis     --           32            --
  (human)
S. enterica 4,5,12:         128          32            --
  i-(human) US clone
B. cereus                   128          32            --
L. monocytogenes 10403S     128          32            --

Microorganism               95%          MBC         Sterile
                          Ethanol   ([micro]l/ml)   distilled
                                     Chloroform       water

E. coli ATCC25822           64           --            --
S. enterica Typhimurium     64           64           >256
  U302 (DT1046)
S. enterica Enteritidis     --           32            --
  (human)
S. enterica 4,5,12:         128          32            --
  i-(human) US clone
B. cereus                   256          256           --
L. monocytogenes 10403S     128          256           --

Table 11. The antibacterial activity (clear zone in mm) of T.
cordifolia crude extracts under five different extractions
condition against six foodborne pathogenic bacteria

Microorganism                95% Ethanol           Chloroform

E. coli ATCC25822         7.75 [+ or -] 0.96   7.25 [+ or -] 0.50
S. enterica Typhimurium   9.00 [+ or -] 1.41   10.50 [+ or -] 2.08
  U302 (DT1046)
S. enterica Enteritidis           --           9.25 [+ or -] 2.06
  (human)
S. enterica 4,5,12:       8.25 [+ or -] 0.50   9.25 [+ or -] 1.26
  i-(human) US clone
B. cereus                 7.25 [+ or -] 0.50   8.75 [+ or -] 1.26
L. monocytogenes 10403S   7.00 [+ or -] 0.82   8.75 [+ or -] 0.96

Microorganism             Hexane        Sterile         Autoclave at
                                       distilled        121[degrees]C
                                         water          15 PSI for 15
                                                           minutes

E. coli ATCC25822           --             --                --
S. enterica Typhimurium     --     7.75 [+ or -] 0.96        --
  U302 (DT1046)
S. enterica Enteritidis     --             --                --
  (human)
S. enterica 4,5,12:         --             --                --
  i-(human) US clone
B. cereus                   --             --                --
L. monocytogenes 10403S     --             --                --

Table 12. The Minimum Inhibitory Concentration (MIC) and Minimum
Bactericidal Concentration (MBC) of T. cordifolia crude extracts
under five different extractions condition against six foodborne
pathogenic bacteria

Microorganism               95%          MIC         Sterile
                          Ethanol   ([micro]l/ml)   distilled
                                     Chloroform       water

E. coli ATCC25822           128          32            --
S. enterica Typhimurium     128          64            256
  U302 (DT1046)
S. enterica Enteritidis     --           64            --
  (human)
S. enterica 4,5,12:         128          64            --
  i-(human) US clone
B. cereus                   128          32            --
L. monocytogenes 10403S     128          32            --

Microorganism               95%          MBC         Sterile
                          Ethanol   ([micro]l/ml)   distilled
                                     Chloroform       water

E. coli ATCC25822           128          32            --
S. enterica Typhimurium     128          64           >256
  U302 (DT1046)
S. enterica Enteritidis     --           64            --
  (human)
S. enterica 4,5,12:         128          64            --
  i-(human) US clone
B. cereus                   256          256           --
L. monocytogenes 10403S     128          128           --
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Article Details
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Author:Yasurin, Patchanee; Piya-Isarakul, Sasiwan
Publication:Journal of Pure and Applied Microbiology
Article Type:Report
Geographic Code:9THAI
Date:Sep 1, 2015
Words:5771
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