Department of Medicine, The Aga Khan University, Stadium Road, Karachi 74800, Pakistan.
Parasitol Res. 2011 Aug;109(2):379-85. doi: 10.1007/s00436-011-2265-z. Epub 2011 Mar 23.
To determine the growth pattern and in vitro susceptibility of Blastocystis hominis to metronidazole (MTZ), garlic, ginger, white cumin, and black pepper. Stool specimens were collected from 16 irritable bowel syndrome (IBS) and 10 controls between July-November 2010. Stool microscopy and culture for B. hominis was performed. Drug susceptibility assays was done using 0.01 and 0.1 mg/ml of MTZ, garlic, ginger, white cumin, and black pepper. Effect was assessed on B. hominis culture after 48 h. Stool DNA was extracted using stool DNA extraction kit (Qiagen) and polymerase chain reaction (PCR) done using subtype-specific sequence-tagged-site primers. B. hominis genotype 3 and coinfection of 1 and 3 tended to grow well in culture compared to isolated type 1 infection. Exposed to MTZ at a concentration of 0.01 mg/ml, 38% (6/16) B. hominis from IBS did not grow in culture compared to 100% (10/10) of B. hominis from control (p = 0.001). When they were exposed to MTZ at 0.1 mg/ml, 56% (9/16) B. hominis from IBS did not grow in cultures compared to 100% (10/10) from control (p = 0.01). Forty-four percent (7/16) B. hominis from IBS did not grow in culture compared to 100% (10/10) B. hominis from control when exposed to garlic at a concentration of 0.01 mg/ml (p = 0.003) and following exposure to garlic at 0.1 mg/ml, 38% (6/16) B. hominis from IBS did not grow in cultures compared to 100% (10/10) from control (p = 0.001). B. hominis isolates from IBS had a cell count of 6,625 at a MTZ concentration of 0.01 mg/ml that reduced to 1,250 as MTZ concentration was increased to 0.1 mg/ml (p = 0.08). B. hominis from IBS with a mean cell count of 3 × 10(5) at baseline decreased to 1 × 10(4) when exposed to garlic at 0.01 mg/ml (p < 0.001) and to 1 × 10(3) (p < 0.001) when garlic was 0.1 mg/ml. B. hominis from IBS cell count decreased to 1 × 10(5) when exposed to white cumin at 0.01 mg/ml (p = 0.01) and to 1 × 10(5) (p < 0.001) when white cumin was 0.1 mg/ml. Exposed to black pepper at 0.1 mg/ml, cell count of B. hominis from IBS decreased to 1 × 10(5) (p = 0.01). B. hominis from IBS decreased to 1.3 × 10(5) exposed to ginger at 0.01 mg/ml (p = 0.001). B. hominis isolates were mostly genotypes 3, type 1 and 3 coinfection, and non-typeable B. hominis isolates. B. hominis isolates from IBS mostly genotype 1 demonstrated an increased sensitivity to garlic at 0.01 mg/ml with a B. hominis cell count of 3,714 compared to 6,142 when exposed to 0.01 mg/ml of MTZ. However, this sensitivity did not increase as garlic concentration was increased to 0.1 mg/ml, for B. hominis cell count was 6,000 compared to 1,428 as MTZ was increased to 0.1 mg/ml.
为了确定人芽囊原虫对甲硝唑(MTZ)、大蒜、生姜、白孜然和黑胡椒的生长模式和体外敏感性,我们于 2010 年 7 月至 11 月期间收集了 16 例肠易激综合征(IBS)和 10 例对照患者的粪便标本。进行粪便显微镜检查和人芽囊原虫培养。使用 0.01 和 0.1mg/ml 的 MTZ、大蒜、生姜、白孜然和黑胡椒进行药物敏感性检测。48 小时后评估对人芽囊原虫培养的影响。使用粪便 DNA 提取试剂盒(Qiagen)提取粪便 DNA,并使用亚类特异性序列标记位点引物进行聚合酶链反应(PCR)。人芽囊原虫基因型 3 和 1 型和 3 型混合感染的菌株比孤立的 1 型感染菌株更容易在培养中生长。在浓度为 0.01mg/ml 的 MTZ 下,与对照中的 100%(10/10)相比,16 例 IBS 患者中的 6 例(6/16)人芽囊原虫在培养中未生长(p=0.001)。当它们暴露在浓度为 0.1mg/ml 的 MTZ 下时,与对照中的 100%(10/10)相比,16 例 IBS 患者中的 56%(9/16)人芽囊原虫在培养中未生长(p=0.01)。当暴露在浓度为 0.01mg/ml 的大蒜下时,与对照中的 100%(10/10)相比,16 例 IBS 患者中的 44%(7/16)人芽囊原虫在培养中未生长(p=0.003),而当暴露在浓度为 0.1mg/ml 的大蒜下时,16 例 IBS 患者中的 38%(6/16)人芽囊原虫在培养中未生长(p=0.001)。与对照中的 100%(10/10)相比,IBS 患者中的人芽囊原虫分离株在 MTZ 浓度为 0.01mg/ml 时的细胞计数为 6625,当 MTZ 浓度增加到 0.1mg/ml 时,细胞计数减少到 1250(p=0.08)。IBS 患者的人芽囊原虫分离株在基线时的平均细胞计数为 3×10(5),当暴露在浓度为 0.01mg/ml 的大蒜下时,细胞计数减少到 1×10(4)(p<0.001),当暴露在浓度为 0.1mg/ml 的大蒜下时,细胞计数减少到 1×10(3)(p<0.001)。IBS 患者中的人芽囊原虫分离株当暴露在浓度为 0.01mg/ml 的白孜然下时,细胞计数减少到 1×10(5)(p=0.01),当暴露在浓度为 0.1mg/ml 的白孜然下时,细胞计数减少到 1×10(3)(p<0.001)。当暴露在浓度为 0.1mg/ml 的黑胡椒下时,IBS 患者中的人芽囊原虫分离株的细胞计数减少到 1×10(5)(p=0.01)。当暴露在浓度为 0.01mg/ml 的生姜下时,IBS 患者中的人芽囊原虫分离株的细胞计数减少到 1.3×10(5)(p=0.001)。IBS 患者中的人芽囊原虫分离株大多为基因型 3、1 型和 3 型混合感染以及非定型人芽囊原虫分离株。IBS 患者中的人芽囊原虫分离株大多为基因型 1,在 0.01mg/ml 的大蒜下显示出对大蒜的敏感性增加,细胞计数为 3714,而在 0.01mg/ml 的 MTZ 下,细胞计数为 6142。然而,当大蒜浓度增加到 0.1mg/ml 时,这种敏感性并没有增加,细胞计数为 6000,而在 MTZ 浓度增加到 0.1mg/ml 时,细胞计数为 1428。