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Occurrence of Potentially Pathogenic Bacterial-Endosymbionts in Acanthamoeba Spp.

Niyyati M, Mafi M, Haghighi A, Hakemi Vala M - Iran J Parasitol (2015 Apr-Jun)

Bottom Line: Sequencing of PCR products confirmed the presence of Pseudomonas aeruginosa and Agrobacterium tumefasiens.The presence of Acanthamoeba bearing pathogenic endosymbionts in water sources leads us to public health issues including improved sanitation and decontamination measures in recreational water sources in order to prevent amoebae-related infection.To the best of our knowledge this is the first report regarding the isolation of A. tumefasiens from Acanthamoeba in Iran and worldwide.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Medical Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ; Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

ABSTRACT

Background: Acanthamoeba- bacteria interactions enable pathogenic bacteria to tolerate harsh conditions and lead to transmission to the susceptible host. The present study was aimed to address the presence of bacterial endosymbionts of Acanthamoeba isolated from recreational water sources of Tehran, Iran. To the best of our knowledge this is the first study regarding occurrence of bacteria in environmental Acanthamoeba spp. in Iran.

Methods: A total of 75 samples of recreational water sources were collected. Samples were cultured on non- nutrient agar 1.5% plates. Positive Acanthamoeba spp. were axenically grown. DNA extraction and PCR reaction was performed using JDP1-2 primers. All positive samples of Acanthamoeba were examined for the presence of endosymbionts using staining and molecular methods. The PCR products were then sequenced in order to determine the genotypes of Acanthamoeba and bacteria genera.

Results: Out of 75 samples, 16 (21.3%) plates were positive for Acanthamoeba according to the morphological criteria. Molecular analysis revealed that Acanthamoeba belonged to T4 and T5 genotypes. Five isolates (35.7%) were positive for bacterial endosymbionts using staining method and PCR test. Sequencing of PCR products confirmed the presence of Pseudomonas aeruginosa and Agrobacterium tumefasiens.

Conclusion: The presence of Acanthamoeba bearing pathogenic endosymbionts in water sources leads us to public health issues including improved sanitation and decontamination measures in recreational water sources in order to prevent amoebae-related infection. To the best of our knowledge this is the first report regarding the isolation of A. tumefasiens from Acanthamoeba in Iran and worldwide.

No MeSH data available.


Related in: MedlinePlus

Cloned Acanthamoeba spp. cysts and trophozoites containing rod shaped-bacteria (magnification x400)
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Figure 2: Cloned Acanthamoeba spp. cysts and trophozoites containing rod shaped-bacteria (magnification x400)

Mentions: Of 16 Acanthamoeba isolate, 14 were cloned successfully. Sequences analysis revealed that Acanthamoeba belonged to T4 and T5 genotypes (Fig. 1) (Table 1). T5 genotype has been isolated from swimming pools. Five isolates (35.7%) of Acanthamoeba were positive for bacterial endosymbionts in their cytoplasm using microscopic observation and gram staining method. Gram staining revealed the presence of bacteria readily in the host amoebae cytoplasm (Fig. 2). Many rod shaped bacteria were localized in the cytoplasm. It should be mention that 4 isolated bacteria showed an approximately 1100 bp band (Fig. 3). However, one strain were failed in sequencing even with repeated PCR reaction. Sequencing of PCR products verified the presence of three Pseudomonas aeruginosa and one Agrobacterium tumefasiens in Acanthamoeba T4 genotypes (Table 1). The result of multialign is shown in Fig. 4 which it shows >5% dissimilarity in 18S rRNA gene. Isolation of Agrobacterium tumefasiens from Acanthamoeba is for the first time worldwide.


Occurrence of Potentially Pathogenic Bacterial-Endosymbionts in Acanthamoeba Spp.

Niyyati M, Mafi M, Haghighi A, Hakemi Vala M - Iran J Parasitol (2015 Apr-Jun)

Cloned Acanthamoeba spp. cysts and trophozoites containing rod shaped-bacteria (magnification x400)
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4522293&req=5

Figure 2: Cloned Acanthamoeba spp. cysts and trophozoites containing rod shaped-bacteria (magnification x400)
Mentions: Of 16 Acanthamoeba isolate, 14 were cloned successfully. Sequences analysis revealed that Acanthamoeba belonged to T4 and T5 genotypes (Fig. 1) (Table 1). T5 genotype has been isolated from swimming pools. Five isolates (35.7%) of Acanthamoeba were positive for bacterial endosymbionts in their cytoplasm using microscopic observation and gram staining method. Gram staining revealed the presence of bacteria readily in the host amoebae cytoplasm (Fig. 2). Many rod shaped bacteria were localized in the cytoplasm. It should be mention that 4 isolated bacteria showed an approximately 1100 bp band (Fig. 3). However, one strain were failed in sequencing even with repeated PCR reaction. Sequencing of PCR products verified the presence of three Pseudomonas aeruginosa and one Agrobacterium tumefasiens in Acanthamoeba T4 genotypes (Table 1). The result of multialign is shown in Fig. 4 which it shows >5% dissimilarity in 18S rRNA gene. Isolation of Agrobacterium tumefasiens from Acanthamoeba is for the first time worldwide.

Bottom Line: Sequencing of PCR products confirmed the presence of Pseudomonas aeruginosa and Agrobacterium tumefasiens.The presence of Acanthamoeba bearing pathogenic endosymbionts in water sources leads us to public health issues including improved sanitation and decontamination measures in recreational water sources in order to prevent amoebae-related infection.To the best of our knowledge this is the first report regarding the isolation of A. tumefasiens from Acanthamoeba in Iran and worldwide.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Medical Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ; Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

ABSTRACT

Background: Acanthamoeba- bacteria interactions enable pathogenic bacteria to tolerate harsh conditions and lead to transmission to the susceptible host. The present study was aimed to address the presence of bacterial endosymbionts of Acanthamoeba isolated from recreational water sources of Tehran, Iran. To the best of our knowledge this is the first study regarding occurrence of bacteria in environmental Acanthamoeba spp. in Iran.

Methods: A total of 75 samples of recreational water sources were collected. Samples were cultured on non- nutrient agar 1.5% plates. Positive Acanthamoeba spp. were axenically grown. DNA extraction and PCR reaction was performed using JDP1-2 primers. All positive samples of Acanthamoeba were examined for the presence of endosymbionts using staining and molecular methods. The PCR products were then sequenced in order to determine the genotypes of Acanthamoeba and bacteria genera.

Results: Out of 75 samples, 16 (21.3%) plates were positive for Acanthamoeba according to the morphological criteria. Molecular analysis revealed that Acanthamoeba belonged to T4 and T5 genotypes. Five isolates (35.7%) were positive for bacterial endosymbionts using staining method and PCR test. Sequencing of PCR products confirmed the presence of Pseudomonas aeruginosa and Agrobacterium tumefasiens.

Conclusion: The presence of Acanthamoeba bearing pathogenic endosymbionts in water sources leads us to public health issues including improved sanitation and decontamination measures in recreational water sources in order to prevent amoebae-related infection. To the best of our knowledge this is the first report regarding the isolation of A. tumefasiens from Acanthamoeba in Iran and worldwide.

No MeSH data available.


Related in: MedlinePlus