title: Occurrence and genetic characterisation of Acanthamoeba spp. from environmental and domestic water sources in Queen Elizabeth Protected Area, Uganda.
abstract: BACKGROUND
  Acanthamoeba is an emerging potentially pathogenic amoeba that has been receiving increasing attention worldwide as a reservoir and potential vector for the transmission of pathogenic bacteria. It is also associated with brain cell damage, keratitis and skin irritation in humans. Its effects are more severe in immunocompromised individuals. This study provides for the first time in Uganda, information on the prevalence and genotypes of Acanthamoeba in environmental and domestic (tap) water.
  
  
  METHODS
  A total of 324 environmental and 84 tap water samples were collected between November 2013 and September 2014. The samples were centrifuged, cultured (Non-Nutrient agar seeded with gram-negative bacteria) and observed under a microscope. After confirmation of Acanthamoeba, genomic DNA was extracted for PCR assays by chemical lysis and purification with phenol/chloroform/isoamyl alcohol. Samples that showed the strongest positive bands (400-600 bp) were subjected to cycle sequencing.
  
  
  RESULTS
  Among environmental and tap water samples, 107 (33 %) and 36 (42.9 %) tested positive for Acanthamoeba spp., respectively. Prevalence of Acanthamoeba from specific environmental locations was as follows; Kazinga channel banks (60.7 %), Fish landing sites (50 %), River Kyambura (39.6 %) and Kazinga mid channel (5.3 %). There was a significant difference (p = 0.001) in the prevalence of Acanthamoeba between sampling sites. The mean (Mean ± SE) occurrence of the organism was higher in Kazinga channel banks (3.44 ± 0.49) and Fish landing sites (3.08 ± 0.53). Correlation between in situ parameters and Acanthamoeba was insignificant except for the Dissolved Oxygen (mg/ML) which was negatively correlated (r = -0.231, p = 0.001) to Acanthamoeba. Six distinct partial Acanthamoeba T-genotype groups T1, T2, T4, T5, T6 and T11 were obtained. Ultimately, Acanthamoeba spp., Acanthamoeba hatchetti and Acanthamoeba polyphaga were isolated in the current study.
  
  
  CONCLUSIONS
  There was a high prevalence of Acanthamoeba in communal piped tap and environmental water used by communities, indicating poor environmental and domestic water quality.
authors.affiliation: Department of Wildlife and Aquatic Animal Resources (WAAR), School of Veterinary Medicine and Animal Resources (SVAR), College of Veterinary Medicine, Animal Resources and Biosecurity (COVAB), Makerere University, P.O. Box 7062, Kampala, Uganda. csente37@gmail.com.
authors.forename: Celsus
authors.initials: C
authors.lastname: Sente
authors.affiliation: Department of Biomolecular Resources and Biolab Sciences, School of Biosecurity, Biotechnical and Laboratory Sciences (SBLS), College of Veterinary Medicine, Animal Resources and Biosecurity (COVAB), Makerere University, P.O. Box 7062, Kampala, Uganda. erujoseph@gmail.com.
authors.forename: Joseph
authors.initials: J
authors.lastname: Erume
authors.affiliation: Department of Wildlife and Aquatic Animal Resources (WAAR), School of Veterinary Medicine and Animal Resources (SVAR), College of Veterinary Medicine, Animal Resources and Biosecurity (COVAB), Makerere University, P.O. Box 7062, Kampala, Uganda. i.naigaga@gmail.com.
authors.forename: Irene
authors.initials: I
authors.lastname: Naigaga
authors.affiliation: Department of Biomolecular Resources and Biolab Sciences, School of Biosecurity, Biotechnical and Laboratory Sciences (SBLS), College of Veterinary Medicine, Animal Resources and Biosecurity (COVAB), Makerere University, P.O. Box 7062, Kampala, Uganda. pjkanywa@gmail.com.
authors.forename: Phillip Kimuda
authors.initials: PK
authors.lastname: Magambo
authors.affiliation: Department of Biomolecular Resources and Biolab Sciences, School of Biosecurity, Biotechnical and Laboratory Sciences (SBLS), College of Veterinary Medicine, Animal Resources and Biosecurity (COVAB), Makerere University, P.O. Box 7062, Kampala, Uganda. ochwosylver@gmail.com.
authors.forename: Sylvester
authors.initials: S
authors.lastname: Ochwo
authors.affiliation: Department of Biomolecular Resources and Biolab Sciences, School of Biosecurity, Biotechnical and Laboratory Sciences (SBLS), College of Veterinary Medicine, Animal Resources and Biosecurity (COVAB), Makerere University, P.O. Box 7062, Kampala, Uganda. mulindwajul@yahoo.com.|Department of Biochemistry and Sports Science, College of Natural Sciences (CONAS), Kampala, Uganda. mulindwajul@yahoo.com.
authors.forename: Julius
authors.initials: J
authors.lastname: Mulindwa
authors.affiliation: Medical Research Council (MRC)/Uganda Virus Research Institute (UVRI), Research Unit on AIDS, P.O. Box 49, Entebbe, Uganda. benignamara@gmail.com.
authors.forename: Benigna Gabriella
authors.initials: BG
authors.lastname: Namara
authors.affiliation: Department of Biomolecular Resources and Biolab Sciences, School of Biosecurity, Biotechnical and Laboratory Sciences (SBLS), College of Veterinary Medicine, Animal Resources and Biosecurity (COVAB), Makerere University, P.O. Box 7062, Kampala, Uganda. katodrago@yahoo.com.
authors.forename: Charles Drago
authors.initials: CD
authors.lastname: Kato
authors.affiliation: Department of Biomolecular Resources and Biolab Sciences, School of Biosecurity, Biotechnical and Laboratory Sciences (SBLS), College of Veterinary Medicine, Animal Resources and Biosecurity (COVAB), Makerere University, P.O. Box 7062, Kampala, Uganda. gsebyatika@gmail.com.
authors.forename: George
authors.initials: G
authors.lastname: Sebyatika
authors.affiliation: Department of Biomolecular Resources and Biolab Sciences, School of Biosecurity, Biotechnical and Laboratory Sciences (SBLS), College of Veterinary Medicine, Animal Resources and Biosecurity (COVAB), Makerere University, P.O. Box 7062, Kampala, Uganda. kevmuwonge@gmail.com.
authors.forename: Kevin
authors.initials: K
authors.lastname: Muwonge
authors.affiliation: Department of Wildlife and Aquatic Animal Resources (WAAR), School of Veterinary Medicine and Animal Resources (SVAR), College of Veterinary Medicine, Animal Resources and Biosecurity (COVAB), Makerere University, P.O. Box 7062, Kampala, Uganda. mocaido@yahoo.com.
authors.forename: Michael
authors.initials: M
authors.lastname: Ocaido
mesh_terms: D000048:Acanthamoeba / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification; D014644:Genetic Variation; D005838:Genotype; D008853:Microscopy; D016133:Polymerase Chain Reaction; D015995:Prevalence; D017422:Sequence Analysis, DNA; D014454:Uganda; D014867:Water / Q000469:parasitology*
chemical_list: D016054:DNA, Protozoan; D014867:Water