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Allicin in vitro killing effect of Acanthamoeba_444

By Henry Watson,2014-10-30 09:24
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Allicin in vitro killing effect of Acanthamoeba_444

Allicin in vitro killing effect of Acanthamoeba

     Abstract Objective To observe the allicin in vitro

    Acanthamoeba in vitro. Methods of Acanthamoeba with different concentrations of allicin 30 ? role in 3,24 h after

    Acanthamoeba was observed under inverted microscope and their morphological changes in cell proliferation rate. The results of the concentration of allicin, respectively 0.01,0.1,1,10 g / L treatment 3 h when Acanthamoeba trophozoites into cysts from, 1,10 g / L observed after treatment 3,24 h a large

    number of cell division debris; cell growth curve compared with the control group, 0.1 g / L group the proliferation rate of normal, while the concentration is higher than 1 g / L above cultured 4 d still can not see cell proliferation.

    Conclusion of allicin in vitro growth of Acanthamoeba significant inhibition and in vitro.

     Key words Acanthamoeba; allicin; in vitro

     The Effect of Allitridium against Acanthamoeba in vitro

     Abstract: ObjectiveTo detect the effect of allitridium on

    Acanthamoeba spp.MethodsAcanthamoeba spp. Was cultured at 30 ? with different concentration of allitridium, the influence of allitridium on the morphological feature, proliferation speed was detected. ResultsAt concentrations of

    0.01, 0.1, 1.0, 10 g / L Allitridium cultured for 3h, the Acanthamoeba trophozoites changed to cysts and in the 10 g / L

    allitridium group cell destroying could be detected. Cell proliferation could not detected.ConclusionAllitridium is effective to destroy Acanthamoeba in vitro.

     Key words: Acanthamoeba; Allitridium; in vitro

     Acanthamoeba (Acanthamoeba) is a widely distributed in the natural environment of the opportunity to pathogenic free-

    living protozoa. Pathogenic Acanthamoeba can cause the body

    granulomatous amoebic encephalitis (Granulomatous Amoebic Encephalitis, GAE), and Acanthamoeba keratitis (Acanhtamoeba Keratitis, AK). Jones, etc., etc. and since the Nagington first reported caused by Acanthamoeba eye infections, along

    with the continuous increase in AK patients, the study of Acanthamoeba caused extensive concern about the scholars. At present reported that at least 26 species have been discovered Acanthamoeba, which often cause human infection is mainly the card's Acanthamoeba (A.caslellanii) and multi-Acanthamoeba

    polyphaga (A.polyphaga).

     Garlic Liliaceae Allium plants, bulbs, has long been considered anti-bacterial, anti-fungal and anti-parasitic

    effects [1], its main bioactive substances are sulfur compounds. Modern medicinal chemistry studies have shown that garlic in the sulfur content of up to 30 kinds, in which diallyl trisulfide generally agreed that the strongest effect, often called allicin. It has been reported to the organization of allicin Entamoeba, Trypanosoma, Giardia Langenbeck,

    Pneumocystis carinii, Cryptosporidium and Toxoplasma gondii to play a certain role in suppression and anti-[1 ~ 3]. The

    experimental observation of allicin in vitro effects of Acanthamoeba protozoa.

     1 Materials and methods

     1.1 Materials Cartesian Acanthamoeba Kyungpook National University in Korea Parasitology Professor Zheng Dongyi available. Allicin Hefong of Shanghai Pharmaceutical Co., Ltd. product, lot number 5E38001.

     1.2 Methods

     1.2.1 reference to the cultivation of Acanthamoeba [4] method of training.

     1.2.2 Morphology of Acanthamoeba logarithmic phase growth of Acanthamoeba cell suspension to adjust the concentration of 2.5 × 105/ml, join the 24-hole cell culture plate in each

    well 1 ml. Experimental groups were each hole to join 0.01,0.1,1,10 g / L allicin 1 ml; the control group received the same amount of PYG medium (proteose peptone-yeast eatract-

    glucose, PYG), 30 ? cultured 3 h, in an inverted observed

    under the microscope of the morphological changes of

    Acanthamoeba.

     1.2.3 the impact on cell growth rate observed with quantitative culture method. Phases will be to adjust the growth of Acanthamoeba cell suspension concentration of 0.5 ×

    105/ml, join the 24-hole cell culture plate, each hole 1 ml. Experimental group were added 0.01,0.1,1,10 g / L allicin 1ml; the control group received the same amount of PYG, each with 4 parallel holes. 30 ? cultured 24 h, the production of

    Acanthamoeba growth curve, and inoculated in solid medium to

    observe the growth of. Reposted elsewhere in the paper for free download http://

     2 Results

     2.1 Morphology of Acanthamoeba Acanthamoeba trophozoites are mostly irregular oval in diameter, mostly 15 ~ 45 μm,

    surface with multiple small thorn-like protuberances of the

    spine, cytoplasmic vacuoles seen in a number; encapsulation type round, diameter 9 ~ 27 μm, there are two layers of cyst

    wall, inner wall was polyhedral type. When the concentration of allicin is equal to or higher than 0.1 g / L culture 3 h,

    the spike-like pseudopodia disappear, forming cysts; 10 g / L group of cells in a small number of cysts observed in a complete, showing that cell debris; and 0.01 g / L group compared with the control group, no significant morphological

    changes.

     2.2 The proliferation rate of Acanthamoeba with different concentrations of allicin role of the 24 h after the culture, production growth curve. The results shown in Figure 1. 6 h after inoculation in control group and 0.01 g / L group can be

    seen the growth of Acanthamoeba, while the other experimental groups were no Acanthamoeba growth; control group, the number of Acanthamoeba within 24 h increased by about 5-fold, while

    the 0.01 g / L group within 24 h of cell growth inhibition, 24

    h later the number increased significantly with the growth trend is almost similar to the control group; 0.01 g / L group after inoculation 24 h, can be seen the proliferation of Acanthamoeba; 1 g / L and 10 g / L Unit 4 d after inoculation still can not see the growth of Acanthamoeba. 30 ? in 24 h

    cultured control group, 0.1 g / ml, 1 g / ml groups, each 50 ml cell suspension centrifuged after fixation, but also to the blank control comparative study found that precipitation cells were significantly different.

     3 Discussion

     Acanthamoeba can cause eye and central nervous system, sub-acute or chronic infection. Acanthamoeba caused GAE often occur in elderly or frail physique because of cancer, chronic wasting disease, or AIDS and other causes of immune function

    in the crowd the opportunity to cause the disease, while the AK often occurs in healthy people, is a serious threat to vision blinding keratopathy.

     Figure 1 Effect of Allicin on Acanthamoeba growth curve (omitted)

     Protozoan Acanthamoeba trophozoites form including the active phase and resting cysts of the two stage, when the drugs act on rapidly when Acanthamoeba trophozoites into cysts. Reported in the literature only in the majority of drugs are killing effect of Acanthamoeba trophozoites, while cysts are less effective. In in vitro experiments have proved PHMB, chlorhexidine, amphotericin, paromomycin, Luo Natamycin, hydroxyethyl sulfonate benzene Amidine propoxyphene, miconazole, as well as to microphone type, etc. [5 ~ 6] Drugs

    Drugs pairs of Acanthamoeba effective, but its effect is not ideal. PHMB and chlorhexidine currently considered the role of the strongest, its most effective role in Acanthamoeba trophozoite concentration of 0.5 ~ 1.0 mg / L, the

    concentration of the effective role of the cyst 1 ~ 5 mg / L [6].

     Garlic Liliaceae Allium plants, bulbs, the main

    components of biologically active substances are sulfur-

    containing compounds learn Diallyl a sulfide, diallyl disulfide, diallyl trisulfide, etc., in which allicin -

    diallyl trisulfide the best. Allicin inhibited the role of some enzymes have an impact on the pathogenic microorganisms, but also through the inhibition of cell proliferation and induce apoptosis in vitro inhibitory effect on a variety of parasites have a good. Lun, etc. [1] reported that allicin on melting histolytica and Giardia inhibition rate of 50% of the

    concentrations of 59 mg / ml and 14 mg / ml. This study shows, 0.1 g / L allicin treatment 24 h, so that Acanthamoeba trophozoites into cysts, and inhibit their proliferation; 1 g / L the role of 3 h can Acanthamoeba trophozoites into a

    package capsule culture 4 d still no recovery, 10 g / L effects can be observed 3 h trophozoites cracking, indicating

    that the concentration is higher than 0.1 g / L when there is an apparent inhibition of cysts, 1 g / L the role of 3 h had a clear in vitro. In this study, different concentrations and time to observe the role of allicin right after Acanthamoeba cell morphology, proliferation rate of change, proved that different concentrations of allicin on the growth of Acanthamoeba protozoa significant inhibition and anti-

    personnel role, and its the role and the role of dose and time-dependent. An effective role in low concentrations of allicin, and has a wide variety of sources, and cheap, less side effects and other characteristics, is promising treatments.

     References

     [1] Lun ZR, Burri C, Menzinger M, et al. Antiparasitic activity of diallyl trisulfide (Dasuansu) on human and animal pathogenic protozoa (Trypanosoma sp., Entamoeba histolytica and Giardia lamblia) in vitro [J]. Ann Soc Belg Med Trop,

    1994,74 (1): 51.

     [2] Rivasi F, Longanesi L, Casolari C, et al. Cytologic diagnosis of Acanthamoeba keratitis. Report of a case with correlative study with indirect immunofluorescence and scanning electron microscopy Acta [J]. Cytol, 1995, 39 (4):

    821 .

     [3] Turner ML, Cockerell EJ. Antigens of selected Acanthamoeba species detected with monoclonal antibodies [J]. Int J Parasitol, 2005, 35 (9): 981.

     [4] Jung Sun Son, Shin Sung-hua, Yuen Ying Hua, et al.

    Acanthamoeba isolation and laboratory culture [J]. Yanbian University Medical Journal, 2003,26 (3): 168.

     [5] Wilhclmus KR. Antiparasitic drugs in Ophthalmology [J]. Int Ophthalmol Clin, 1996, 36 (2): 117.

     [6] Kbunkitti W, Lloyd D, Fure JR, et al.The lethal

    effects of biguanides on cysts and trophozoites of Acanthamoeba castellanii [J]. J Appl Bacteriol .1996, 81 (1): 73. Reposted elsewhere in the paper for free download Center http://

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