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Aim: The aim of the present study was to simultaneously investigate parasitic contamination of treated wastewater and downstream vegetable farms that are irrigated with treated sewage, during a year
Background: [Oo] Cysts and eggs of parasites are resistant to most of routine wastewater treatment process. Irrigation of vegetables farms with either treated wastewater or illegally use of raw wastewaters enhances the risk of contamination with enteric pathogens
Methods: The treated wastewater samples were taken after chlorination from a wastewater treatment plant located at the south of Tehran. In addition, 60 vegetable samples [5 samples from each farm] were collected from the selected downstream farms that routinely used treated wastewater for irrigation of crops. Parasitological tests were performed using Ziehl-Neelsen, conventional lugol's iodine staining and direct microscopical examination
Results: Parasites including free living larvae, eggs of Toxoascaris leonina, egg of Toxocara sp. Trichuris sp, Trichostrongylus sp and amoeboid trophozoite were seen in 5/12 [41.7%] of vegetable samples gathered during a year. There was no statistically significant correlation between the season and parasitic contamination of the vegetables [P= 1]. Furthermore, parasitic contamination was observed in 7/12 [53.8%] of treated wastewater samples. The correlation between season and parasitic contamination of treated wastewater was evaluated that the results showed a higher contamination of treated wastewater in spring and autumn [P<0.05].Fisher's exact test also showed that there was no significant correlation between parasitic contaminations of vegetable samples and treated wastewater according to seasonal change
Conclusion: The results showed parasites in both treated wastewater plant and downstream crops farms that suggests the public health importance of the quality of water resources that routinely used for irrigation of vegetable farms
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OBJECTIVE@#To verify phylogeography and genetic structure of Acanthamoeba populations among the Iranian clinical isolates and natural/artificial environments distributed in various regions of the country.@*METHODS@#We searched electronic databases including Medline, PubMed, Science Direct, Scopus and Google Scholar from 2005 to 2016. To explore the genetic variability of Acanthamoeba sp, 205 sequences were retrieved from keratitis patients, immunosuppressed cases and environmental sources as of various geographies of Iran.@*RESULTS@#T4 genotype was the predominant strain in Iran, and the rare genotypes belonged to T2, T3, T5 (Acanthamoeba lenticulata), T6, T9, T11, T13 and T15 (Acanthamoeba jacobsi). A total of 47 unique haplotypes of T4 were identified. A parsimonious network of the sequence haplotypes demonstrated star-like feature containing haplogroups IR6 (34.1%) and IR7 (31.2%) as the most common haplotypes. In accordance with the analysis of molecular variance, the high value of haplotype diversity (0.612-0.848) of Acanthamoeba T4 represented genetic variability within populations. Neutrality indices of the 18S ribosomal RNA demonstrated negative values in all populations which represented a considerable divergence from neutrality. The majority of genetic diversity belonged to the infected contact lens and dust samples in immunodeficiency and ophthalmology wards, which indicated potential routes for exposure to a pathogenic Acanthamoeba sp. in at-risk individuals. A pairwise fixation index (F) was from low to high values (0.02433-0.41892). The statistically F points out that T4 is genetically differentiated between north-west, north-south and central-south metapopulations, but not differentiated between west-central, west-south, central-south, and north-central isolates.@*CONCLUSIONS@#An occurrence of IR6 and IR7 displays that possibly a gene flow of Acanthamoeba T4 occurred after the founder effect or bottleneck experience through ecological changes or host mobility. This is the first systematic review and meta-analysis providing new approaches into gene migration and transmission patterns of Acanthamoeba sp, and targeting at the high-risk individuals/sources among the various regions of Iran.
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Objective To verify phylogeography and genetic structure of Acanthamoeba populations among the Iranian clinical isolates and natural/artificial environments distributed in various regions of the country. Methods We searched electronic databases including Medline, PubMed, Science Direct, Scopus and Google Scholar from 2005 to 2016. To explore the genetic variability of Acanthamoeba sp, 205 sequences were retrieved from keratitis patients, immunosuppressed cases and environmental sources as of various geographies of Iran. Results T4 genotype was the predominant strain in Iran, and the rare genotypes belonged to T2, T3, T5 (Acanthamoeba lenticulata), T6, T9, T11, T13 and T15 (Acanthamoeba jacobsi). A total of 47 unique haplotypes of T4 were identified. A parsimonious network of the sequence haplotypes demonstrated star-like feature containing haplogroups IR6 (34.1%) and IR7 (31.2%) as the most common haplotypes. In accordance with the analysis of molecular variance, the high value of haplotype diversity (0.612–0.848) of Acanthamoeba T4 represented genetic variability within populations. Neutrality indices of the 18S ribosomal RNA demonstrated negative values in all populations which represented a considerable divergence from neutrality. The majority of genetic diversity belonged to the infected contact lens and dust samples in immunodeficiency and ophthalmology wards, which indicated potential routes for exposure to a pathogenic Acanthamoeba sp. in at-risk individuals. A pairwise fixation index (F
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Background: Free-living amoebae have various genera that are found in several environmental niches such as soil, freshwater, dust, seawater and hotsprings. Most of Free-living amoebae are normally harmless to humans. However, some ameoba such as Acanthamoeba and also Naegleria fowleri, Balamuthia manderillaris and Sappinia are identified as opportunistic and pathogenic amoebae that can cause eye diseases, encephalitis, and meningoencephalitis in human. Vannellidae are a family of free-living amoebae and exist mainly in soil, freshwater, and marine habitats. This amoeba is nonpathogenic for human, but can act as a Trojan horse for other pathogens such as Microsporidia. The present study reports the occurrence of Vannella spp. in a hotspring of Amol city
Materials and Methods: 22 samples were taken from hotsprings of Mazandaran province during our previous study. The plates were checked for the presence of Vannella spp. according to the specific morphological criteria. DNA extraction, PCR and sequencing was performed on the positive isolate
Results: The result showed that one plate contained fan-shaped amoebae suspected to Vannella spp. PCR analysis and sequencing was confirmed the occurrence of Vannella spp. in one sample of a hot spring of Amol, northern Iran
Conclusion: The result confirmed the presence of Vannella amoebae in the hotspring of Amol city. More studies are needed to clarify the real distribution of Vannella spp. in environmental niches and its pathogenic potential in Iran and worldwide
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Background and Objectives: Free-living amoebae [FLA] include various genera habitat in water sources; some FLA can lead to severe complications in high risk people. The present study aimed to isolate free living amoebae using morphological methods in recreational water sources of Tehran
Materials and Methods: This cross sectional study was performed during 8 months in 2014. Seventy five samples from Tehran were collected and filtered. Samples were cultured in 1.5% non-nutrient bacto-agar. Plates were then monitored for the presence of amoebae daily and positive plates were cloned. In the present study, identification was based on morphological criteria and page key. The page key is based on morphological character of free living amoebae such as trophozoites shape, pseudopodia shape, amoebae nucleus, and endo and ecto-cysts in the cystic form, These criteria could lead to identification of amoeba at the family and genus level
Results: Out of 75 water samples, 18 [24%] were positive for free living amoebae. Of 40 pond waters, 13 [32,5%] were positive including Acanthamoeba, Hartmcmnella and Vahlkampfiids [Naegleria] and out of 35 samples 5 [14.2%] strain belonging io Acanthamoeba were identified based on morphological criteria
Conclusion: According to the presence of free living amoebae in recreational water sources, it is necessary to alert swimming pools authorities and high risk people. Additionally, posting alarming signs and educating the high risk people is of utmost importance to prevent free living amoebae-related infections
Subject(s)
Swimming Pools , Parks, Recreational , Ponds/microbiology , Cross-Sectional StudiesABSTRACT
Background: Free-living amoebae [FLA] are group of protozoa distributed worldwide in many habitats mainly water and soil related sources. Some members of FLA are able to act as opportunistic pathogens and are environmental carriers of other pathogenic agents such as bacteria and viruses. Vermamoeba vermiformis is a highly abundant FLA species in water bodies and has recently gained environmental importance as it acts as a vehicle of many pathogenic bacteria such as Legionella pneumophila
Cases Report: In this study, water samples were collected from the island of El Hierro, Canary Islands, Spain during 2015. El Hierro island was designated by UNESCO as a biosphere reserve and it is currently the less populated of the Canary Islands. The water samples were culture on 2 % Non-Nutrient Agar [NNA] plates covered with a thin layer of heat killed E. coli and checked daily for the presence of FLA. After a week, V. vermiformis amoebae were observed in the plates incubated at room temperature and 37 [degree]C. Molecular characterization was carried out by amplifying the 18S rDNA gene and DNA sequencing, confirming that the isolated strain belonged to Vermamoeba vermiformis species
Conclusion: To the best of our knowledge, this is the first report of Vermamoeba vermiformis isolation in the island of El Hierro and the second report of this species in the Canary Islands
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In the latest articles which were published during 2013-2014, Trichomonas vaginalis [T. vaginalis] was mentioned as a neglected sexual transmission disease [STD], while the exact mechanism of its pathogenesis has not been cleared yet. Although trichomonasiasis is easy curable, there is concern that resistance to drug are increasing. This common infection as concerning the important public health implications needs more research to be done for understanding the diagnosis, treatment, immunology and pathogenesis. In this review we searched all valuable and relevant information considering the pathogenesis of T. vaginalis. We referred to the information databases of Medline, PubMed, Scopus and Google scholar. The used keywords were the combinations of T. vaginalis and words associated with pathogenicity. This review discusses the host-parasite interaction and pathogenicity of this parasite
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Acanthamoeba spp. is the causative agent of blindness keratitis and fatal encephalaitis. Presence of Acanthamoeba spp. in a wide variety of niches such as different water types can lead to exposure of high risk people such as contact lens wearers. The main aim of the present study was to explore the occurrence of Acanthamoeba genotypes in the recreational water sources using both morphological and molecular approaches in Gilan province, Iran. Overall, 50 samples were collected from recreational water sources including man- made and natural waters in Gilan province. Filtration and cultivation of samples was performed using non-nutrient agar. Cloning of Acanthamoeba spp. was done to eliminate bacterial and fungi contamination. PCR amplification and sequencing were performed using genus-specific primer pair. Genotype identification was based on homology analysis of 18S rRNA gene [DF3] of the obtained sequences with the available genes in the gene bank data base. Out of 50 water samples, 15 [30%] were positive for Acanthamoeba trophozoites and cysts according to morphological criteria. Cloning of 13 isolates [26%] was done successfully. Molecular analysis of 13 Acanthamoeba strain revealed that all isolates were belonged to potentially pathogenic T4 genotype. T4 genotype is the main cause of Acanthamoeba-related infections. Presence of Acanthamoeba belonged to T4 genotype in recreational water sources is of concern for high risk people. Alarming sign and education to high risk people is of utmost importance to prevent such infections
Subject(s)
Genotype , WaterABSTRACT
Background: Acanthamoeba keratitis [AK] is potentially a sight-threatening infection and its treatment is challenging. This is mainly due to presence of resistant cyst form. Indeed, cysts are highly resistant to current available drugs. Chemical drugs are toxic to human keratocytes. It should also be mentioned that most available anti-Acanthamoeba drugs are poorly cysticidal, In Iran and worldwide, AK cases continue to rise and therefore, novel effective drugs are urgently needed for the treatment of AK.
Materials and Methods: In the present study, the in vitro activity of serial dilutions [10, 15, 20 and 25 mg/mL] of chloroformic fractions including primary chloroformic fraction [minimum amount of chloroform], middle chloroformic fraction and remaining chloroformic fraction [most amount of chloroform] of Trigonella foenum graecum seed were evaluated against Acanthamoeba trophozoites and cysts. Cytotoxic assay of fractions at different concentrations [25, 50, 100, 200, 300, 400, 500 mg/ml] of test material was identified on mice Macrophage cells using MTT method.
Results: The obtained results revealed that the tested fractions presented anti-amoebic activities in a time and dose dependent cycle. Anti-Acanthamoeba activity of remaining chloroformic fraction was more than other fractions. Trophozoites/cysts were eliminated when incubated with 15 and 20 mg/ml concentrations of remaining chloroformic fraction after 24 hours. Viability of macrophage cells was noted 100 % with 25 and 50 mg/ml concentration of remaining chloroformic fraction. Our results indicate that the plant fractions are safe for mammalian cells.
Conclusion: Further studies should be performed in order to detect the active chemical compounds which could be used for the development of novel therapeutic approaches against Acanthamoeba infections. To the best of our knowledge, this is the first study on the activity of chloroformic fractions of Trigonella foenum graecum [seed] against Acanthamoeba spp.
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Background: Free-living amoebae [FLA] are protozoa that are widely distributed in the environment mainly in water and soil related habitats. Thermophilic amoebae are among the most common FLA in water bodies, being Vermamoeba vermiformis one of the most common species reported worldwide from these sources. Interestingly, V. vermiformis has often been reported to survive at high temperatures and osmotic pressure worldwide.
Materials and Methods: In this study, snow samples were collected from Mount Teide, Tenerife, Canary Islands during the winter season of 2014. The samples were culture on non nutrient agar plates and checked daily for the presence of FLA. After a week, V. vermiformis amoebae were observed in the plates incubated at room temperature and 37ºC.
Results: Molecular characterization was carried out by amplifying the 18S rDNA gene and DNA sequencing, confirmed that the isolated strain belonged to Vermamoeba vermiformis species.
Conclusion: To the best of our knowledge, this is the first report of Vermamoeba vermiformis isolation from such an inusual habitat [low temperatures and high altitude] and the first report of these species in the Canary islands.
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Acanthamoeba-related disease have a poor prognosis according to many previous studies. Thus researches regarding biochemical and molecular aspects of this organism are a high priority. To this end achieving high amount of amoebae in culture is the first step for such studies. The main aim of the present research was to address the usage of TYI-S-33 [Tripticase, Yeast extract, iron-serum] medium as an enrichment component for achieving high and fast growth of trophozoites in agar culture medium within 24 hours. Overall, 10 Acanthamoeba strains were cultured, cloned and genotyped and the cultures were then enriched with addition of TYI-S-33 medium. Amoebae growth was then monitored daily. Ten plates also were used without addition of TYI-S-33 medium. The result of the present research revealed that addition of TYI-S-33 medium is a promising approach for obtaining 100% trophozoites within 24 hours of culture. To the best of our knowledge this is the first report of successful achieving high amount of trophozoites within short time that able researchers to arrange molecular and biochemical assays
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Free-living amoebae [FLA] including Acanthamoeba spp. and Hartmannella spp. are the causative agents of serious corneal infection especially within contact lens wearers. Thus contact lenses and their storage case could be a suitable niche for potentially pathogenic amoebae. The main objective of the present study was to evaluate the contamination of contact lenses to free living amoebae using morphological and sequencing based methods. Overall, 90 volunteers provided their contact lenses. All volunteers wore soft contact lenses. Both lenses were cultured in the same plate. Forty-eight of the volunteers were medical and dentistry student and 42 were ophthalmology attendees of hospitals in Tehran, Iran. All of the samples were inoculated to non-nutrient medium and monitored daily for the outgrowth of the amoebae. PCR and sequencing were performed using various primer pairs. Of the 90 volunteers, 9 [10%] were positive for free-living amoebae outgrowth. Morphological analysis revealed that 3 isolates were belonged to Hartmannella genus according to small round cysts and 6 isolates were belonged to Acanthamoeba genus based on the star shape of endocysts. Sequencing revealed that Acanthamoeba belonged to T4, T3 and T5 genotype. Hartmannella were also belonged to vermiformis species. The presence of potentially pathogenic free living amoebae including Acanthamoeba and Hartmannella could be a high risk for people using soft contact lenses. These results revealed that improved clarification and professional recommendations for contact lens wearers is of utmost importance
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Poor hygiene will provide good condition for corneal infections by opportunistic free-living amoebae [FLA] in soft contact lens wearers. In the present study an amoebic keratitis due to Hartmannella has been recognized in a 22-year-old girl with a history of improper soft contact lens use. She had unilateral keratitis on her left eye. Her clinical signs were eye pain, redness, blurred vision and photophobia. The round cysts of free-living amoebae were identified in non-nutrient agar medium by light microscopy. These cysts were suspected to be Hartmannella using morphological criteria. A PCR assay has been confirmed that the round cysts were belonged to H. vermiformis