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Acanthamoeba

Q: What are the different types of Acanthamoeba?

Acanthamoeba is a genus of free-living amoebae that includes several species. The most common and clinically relevant species are Acanthamoeba castellanii, Acanthamoeba culbertsoni, Acanthamoeba polyphaga, and Acanthamoeba hatchetti. These species can cause serious infections like Acanthamoeba keratitis and granulomatous amoebic encephalitis.

Acanthamoeba: A genus of free-living amoebae found in soil, water, and air.
Certain species can cause serious infections, such as Acanthamoeba keratitis, a painful and potentially blinding eye infection, and granulomatous amoebic encephalitis, a rare but often fatal brain infection.
These amoebae are opportunistic pathogens that thrive in immunocompromised individuals.
Effective treatment and prevention strategies are crucial for managing Acanthamoeba-related diseases.
Resaerch in this area is vital to enhancing our understanding and improving patient outcomes.

Most cited protocols related to «Acanthamoeba»

Fungal Corneal Ulcer Treatment Protocol

Eligible patients had a smear-positive fungal corneal ulcer and baseline visual acuity of 20/40 (0.3 logMAR) to 20/400 (1.3 logMAR) (Table 1). Reasons for exclusion included impending perforation, evidence of bacterial, Acanthamoeba, or herpetic keratitis, being younger than 16 years, and bilateral ulcers or visual acuity worse than 20/200 (1.0 logMAR) in the nonaffected eye. Masked assignment to the treatment intervention was performed after determination of eligibility and consent to participate. Enrollment centers included the Aravind Eye Care System in India (Madurai, Pondicherry, and Coimbatore) and the Francis I. Proctor Foundation, University of California, San Francisco.

Prajna N.V., Krishnan T., Mascarenhas J., Rajaraman R., Prajna L., Srinivasan M., Raghavan A., Oldenburg C.E., Ray K.J., Zegans M.E., McLeod S.D., Porco T.C., Acharya N.R, & Lietman T.M. (2013). The Mycotic Ulcer Treatment Trial: A Randomized Trial Comparing Natamycin vs Voriconazole. JAMA ophthalmology, 131(4), 422-429.

Publication 2013

Acanthamoeba Bacteria BAD protein, human Corneal Ulcer Eligibility Determination Keratitis, Herpetic Patients Ulcer Visual Acuity Youth

Multifaceted Genome Assembly Approach

Genome assembly was carried out using a two-step process. Firstly, the Illumina reads were assembled using the Velvet [96 (link)] short read assembler to generate a series of contigs. These assembled contigs were used to generate a set of pseudo-reads 400 bp in length. These pseudo reads were then assembled in conjunction with the 454 FLX and Sanger sequences using version 2.3 of the GS De Novo Assembler using default parameters (Table S1.1.1 in Additional file 1). The assembly contained 45.1 Mb of scaffold sequence, of which 3.4 Mb (7.5%) represents gaps and 75% of the genome is contained in less than 100 scaffolds. For assembly statistics see Table S1.2.1 in Additional file 1. In order to determine the coverage of the transcriptome, we aligned our genome assembly to a publicly available EST dataset from GenBank (using the entrez query acanthamoeba EST) AND 'Acanthamoeba castellanii' [porgn:txid5755]). Of the 13,784 EST sequences downloaded, 12,975 (94%) map over 50% of their length with an average percent identity of 99.2% and 12,423 (90%) map over 70% of their length with an average percent identity of 99.26%.

Clarke M., Lohan A.J., Liu B., Lagkouvardos I., Roy S., Zafar N., Bertelli C., Schilde C., Kianianmomeni A., Bürglin T.R., Frech C., Turcotte B., Kopec K.O., Synnott J.M., Choo C., Paponov I., Finkler A., Heng Tan C.S., Hutchins A.P., Weinmeier T., Rattei T., Chu J.S., Gimenez G., Irimia M., Rigden D.J., Fitzpatrick D.A., Lorenzo-Morales J., Bateman A., Chiu C.H., Tang P., Hegemann P., Fromm H., Raoult D., Greub G., Miranda-Saavedra D., Chen N., Nash P., Ginger M.L., Horn M., Schaap P., Caler L, & Loftus B.J. (2013). Genome of Acanthamoeba castellanii highlights extensive lateral gene transfer and early evolution of tyrosine kinase signaling. Genome Biology, 14(2), R11.

Publication 2013

Acanthamoeba Acanthamoeba castellanii BP 400 Genome Transcriptome

Construction of Fluorescent Plasmid Reporters

Protocol full text hidden due to copyright restrictions

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Publication 2009

Acanthamoeba Base Pairing Codon, Initiator GAPDH protein, human Genes Genome Methionine Plasmids Reading Frames

Isolation of Novel Amoeba-Associated Viruses

Acanthamoeba castellanii (Douglas) strain Neff (ATCC 30010) cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and cultured in peptone-yeast-glucose (PYG) medium at 26°C as described previously (14 (link), 81 (link)). An outflow water/soil sample (50 ml) was collected from a water sample spilled out from a hot spring in Japan. The water temperature was 43.4°C at the sampling site. After removal of floating bacteria and small viruses by filtration using a 20-μm filter (no. 43; Whatman International, Maidstone, UK), the collected mud and dead leaves were resuspended in 13 ml of sterile phosphate-buffered saline (PBS) and stirred gently for 1 day at room temperature. The sample was again filtered through another 20-µm filter. Then, the filtered sample (10 ml) was further filtered through a sterile 1.2-µm filter (Millex-AA; Merck Millipore, Darmstadt, Germany). The filtrate (9.5 ml) was then mixed with PYG medium (18 ml). Acanthamoeba cell suspension (0.5 ml) was added and incubated with gentle stirring for 1 h at room temperature, followed by incubation at 26°C in a total of 142 wells using two 96-well microplates. After 5 days, amoeba cells with delayed proliferation were screened. Culture supernatant from growth-retarded wells showing phenotypical difference was inoculated into fresh amoeba cells in an individual well of a 12-well microplate. After 3 days, supernatant of all three wells with cell encystment was inoculated into a fresh amoeba cell suspension in three 25-cm² culture flasks and then in three 75-cm² culture flasks. Supernatant from each 75-cm² culture flask was stored at 4°C as an isolated virus solution (named HS-1, HS-2, and HS-3).

Yoshikawa G., Blanc-Mathieu R., Song C., Kayama Y., Mochizuki T., Murata K., Ogata H, & Takemura M. (2019). Medusavirus, a Novel Large DNA Virus Discovered from Hot Spring Water. Journal of Virology, 93(8), e02130-18.

Publication 2019

Acanthamoeba Acanthamoeba castellanii Amoeba Bacteria Cell Proliferation Cells Filtration Glucose Peptones Phosphates Saline Solution Sterility, Reproductive Virus Yeast, Dried

Isolation and Preparation of hMDMs and U937 Cells

Isolation and preparation of the human monocyte-derived macrophages (hMDMs) from peripheral blood of volunteers and macrophage-like U937 cells was carried out as previously described (Doyle et al., 2001 (link); Santic et al., 2005 (link); Tilney et al., 2001 (link)). Cells were maintained in RPMI-1640 tissue culture medium (Mediatech) supplemented with 10% heat-inactivated fetal bovine serum FBS (Atlas Biologicals). Axenic Acanthamoeba polyphaga was cultured as adherent cells in PYG medium as previously described (Gao et al., 1997 (link)). All cells were grown at 37°C in presence of 5% CO₂.

Al-khodor S., Price C.T., Habyarimana F., Kalia A, & Kwaik Y.A. (2008). A Dot/Icm-translocated ankyrin protein of Legionella pneumophila is required for intracellular proliferation within human macrophages and protozoa. Molecular microbiology, 70(4), 908-923.

Publication 2008

Acanthamoeba Biological Factors BLOOD Cells Cultured Cells Homo sapiens isolation Macrophage Tissues U937 Cells Voluntary Workers

Most recents protocols related to «Acanthamoeba»

Imaging Acanthamoeba Disinfection with Contact Lens Solutions

Similar to our previous description [8 (link)], sterile aluminum transmission flow cells (Biosurface Technologies Corporation, Bozeman, MT, USA) were assembled with glass coverslips (Figure 1). Acanthamoeba suspensions (3.0 × 10⁴ cells/mL) in one-quarter Ringer’s solution were added through the ports of the flow cells. Each flow cell was placed on the microscope stage and kept there for the duration of the experiment, to reduce physical interference with Acanthamoeba behavior. The flow cell ports were clamped shut to stop the flow of the Acanthamoeba suspension, and Acanthamoeba trophozoites were allowed to adhere to glass coverslips for one hour. Following this, flow cell solutions were either maintained as one-quarter Ringer’s solution, or changed to one of the four contact lens care solutions (CLCs). To exchange fluids, 4 mL of a solution was slowly added through the ports of the flow cell, taking care not to disturb the adhered amoebae. The flow cell ports were again clamped to prevent fluidic movement, and amoebae were imaged for 6 h (6 h was used to create identical testing conditions, as the majority of the solutions used specify a 6-h disinfection time, although PAPB/PQ requires only 4 h). After 6 h, all solutions were exchanged for axenic culture media (AC6) to determine the ability of Acanthamoebae to recover following disinfection. The AC6 condition was imaged for a further 12 h, for a total experimental imaging time of 18 h. Images were taken at 4× magnification using a Nikon Eclipse Ti-U Microscope (Nikon, Tokyo, Japan), every 24 s. The images were subsequently combined into a video format using NIS Elements AR 3.2.

Campolo A., Patterson B., Lara E., Shannon P, & Crary M. (2023). Complete Recovery of Acanthamoeba Motility among Surviving Organisms after Contact Lens Care Disinfection. Microorganisms, 11(2), 299.

Publication 2023

Acanthamoeba Aluminum Amoeba Cells CLC protein, human Culture Media Disinfection Microscopy Movement Physical Examination polyaminopropyl biguanide Ringer's Solution Sterility, Reproductive Transmission, Communicable Disease Trophozoite

Culturing and Characterizing Acanthamoeba Strains

As previously described [8 (link),10 (link)], trophozoites were axenically cultured in AC6 medium (axenic culture medium; containing 20 g biosate peptone, 5 g glucose, 0.3 g KH₂PO₄, 10 µg vitamin B12, and 5 mg L-methionine per liter of distilled deionized water). AC6 was adjusted to pH 6.6–6.95 with 1M NaOH and autoclaved at 121 °C for 20 min, before being stored at room temperature for use within 3 months. Organisms were harvested using one-quarter Ringer’s solution. Acanthamoeba strains were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). Acanthamoeba polyphaga (ATCC 30461), Group T4, isolated from a human eye infection (Namibia or South Africa, 1973) and Acanthamoeba castellanii (ATCC 50370), also Group T4, isolated from a human eye infection (New York, NY, 1978) were the two clinical strains used in this study. Importantly, these two commonly used clinically relevant strains belong to the T4 genotype, which is the genotype most frequently associated with Acanthamoeba keratitis [11 (link),12 (link),13 (link)]. To create a homogenous population of Acanthamoeba trophozoites, amoebae were scaled up in fresh AC6 media 24 h prior to testing. The multi-purpose solutions tested were chosen for their representation of popular multi-purpose solutions, are identified by biocide throughout the manuscript, and can be found in Table 1.
In order to simulate the common practice of patients of re-using old solutions, cycled PHMB was created by filling the manufacturer-provided contact lens cases with PHMB. Lenses were placed in cases overnight for 12 to 16 h and then removed. This was repeated for 7 days without replacing or refreshing the CLC, and the resulting CLC was collected.

Publication 2023

Acanthamoeba Acanthamoeba castellanii Acanthamoeba Keratitis Amoeba Axenic Culture Biocides Cobalamins Contact Lenses Eye Infection Genotype Glucose Homo sapiens Homozygote Lens, Crystalline Methionine Patients Peptones Ringer's Solution Strains Trophozoite

Evaluating PCR Assay Specificity for Abortion Pathogens

Specificity of the PCR assays was tested using DNA extracted from different non-target bacteria often associated with products of abortion. Live intracellular control strains C. abortus (ATCC VR-165), P. acanthamoeba (ATCC VR-1476) and W. chondrophila (ATCC VR-1470) were procured from the American Type Culture Collection, Manassas, VA, USA. These bacteria were cultivated in five millilitres serum-casein-glucose-yeast extract medium (SCGYEM) at 30 °C for 6 days with Acanthamoeba castellani (ATCC 50739) as host according to ATCC instructions. Cultures were then harvested and centrifuged at 5000× g for 5 min. The supernatant was discarded and the pellet was resuspended in one millilitre of sterile PCR grade water.
Live bacterial control strains that grow on acellular media (Table 1) were streaked on 5% blood agar plates and incubated in 5% CO₂ at 37 °C until growth was observed. Bacterial suspensions were made in 5% saline to a turbidity equal to a 0.5 McFarland standard. Fifty microlitres of Acholeplasma laidlawlii (NCTC 10116) suspension was inoculated in Mycoplasma broth (Oxoid CM0403 & SR0059, Thermoscientific, Gauteng, South Africa) and incubated in normal air at 37 °C until the broth was turbid. The bacterial suspensions and Acholeplasma laidlawlii broth culture were then diluted 1:100 in sterile PCR grade water.

Jonker A, & Michel A.L. (2023). Optimization and Application of Real-Time qPCR Assays in Detection and Identification of Chlamydiales in Products of Domestic Ruminant Abortion. Pathogens, 12(2), 290.

Publication 2023

Acanthamoeba Acanthamoeba castellanii Acholeplasma Agar Bacteria Biological Assay Blood Caseins Fetuses, Aborted Glucose Induced Abortions Mycoplasma Protoplasm Saline Solution Serum Sterility, Reproductive Strains Yeast, Dried

Chlamydial Pathogen Detection Limits

Samples of stomach content, placenta and lung from cases where no placentitis or pneumonia was reported were analysed with the qPCRs in this study. DNA from samples where there was no amplification of target DNA was spiked with 10, 5, 2 and 1 DNA copies (C. abortus/C. pecorum & P. acanthamoeba/W. chondrophila) or 1000, 500, 250 and 62 DNA copies (Pan-Chlamydiales) of the synthetic controls (Integrated DNA Technologies, Coralville, IA, USA). Six repetitions of the spiked samples were analysed to determine detection limits of the singleplex and duplex qPCR assays.

Publication 2023

Acanthamoeba Biological Assay Chlamydiales Fetuses, Aborted Lung Placenta Pneumonia Stomach Contents

Chlamydiales Detection Multiplex qPCR

Primers and probes were selected from published studies according to base pair size and annealing temperature [27 (link),32 (link),34 (link),35 (link),36 (link)]. The probes for Pan-Chlamydiales, W. chondrophila and P. acanthamoeba were slightly modified by replacing locked nucleic acids by a Minor Groove Binder (MGB) probe. Primers were synthesised by Integrated DNA Technologies, Coralville, IA, USA (www.idtdna.com, accessed on 20 November 2017) and probes by Thermo Fisher Scientific, Waltham, MA, USA (www.thermofischer.com, accessed on 13 November 2017). Sequences and targets for primers and probes are as in Table 2.
The Pan-Chlamydiales qPCR was used as a singleplex assay. The C. abortus and C. pecorum qPCRs were combined in a duplex qPCR assay, while the P. acanthamoeba and W. chondrophila qPCRs were combined in a second duplex qPCR assay.

Publication 2023

Acanthamoeba Base Pairing Biological Assay Chlamydiales Fetuses, Aborted locked nucleic acid Oligonucleotide Primers

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What are the different types of Acanthamoeba?

How can PubCompare.ai help with Acanthamoeba research?

PubCompare.ai allows you to screen protocol literature more efficiently and leverage AI to pinpoint critical insights. The platform can help researchers identify the most effective protocols related to Acanthamoeba for their specific research goals. PubCompare.ai's AI-driven analysis can highlight key differences in protocol effectiveness, enabling you to choose the best option for reproducibility and accuracy in your Acanthamoeba studies.

What are the common challenges in working with Acanthamoeba?

Working with Acanthamoeba can present several challenges, such as maintaining optimal growth conditions, accurately identifying different species, and developing effective treatment strategies. Acanthamoeba can be difficult to culture and may require specialized media and techniques. Additionally, Acanthamoeba infections can be tricky to diagnose, as the symptoms can be similar to other eye or brain infections. Leveraging PubCompare.ai can help researchers overcome these challenges by identifying the most reliable and effective protocols from the literature.

What are some specific applications of Acanthamoeba research?

Acanthamoeba research has a range of applications, including: 1) Studying the pathogenesis and virulence mechanisms of Acanthamoeba infections, which can lead to improved diagnostic and treatment methods. 2) Investigating the ecology and environmental distribution of Acanthamoeba, which can inform prevention strategies. 3) Exploring the potential use of Acanthamoeba in bioremediation and wastewater treatmetn. 4) Utilizing Acanthamoeba as a model organism for cell biology and immunology reseacrh. PubCompare.ai can help researchers navigate the vast literature to identify the most relevant protocols for their specific Acanthamoeba applications.

How can researchers ensure the accuracy and reproducibility of their Acanthamoeba experiments?

Ensureing the accuracy and reproducibility of Acanthamoeba experiments is crucial. Some key strategies include: 1) Carefully following standardized protocols for Acanthamoeba culture and identification. 2) Utilizing robust quality control measures, such as regular microscopic examinations and viability assays. 3) Comparing results across multiple independent experiments and replicates. 4) Benchmarking findings against established Acanthamoeba research. PubComapre.ai can assist in this process by helpnig researchers identify the most reliable and effective protocols from the literature, enabling them to optimize their experimental design and improve the overall quality of their Acanthamoeba research.

More about "Acanthamoeba"

Acanthamoeba, a genus of ubiquitous free-living amoebae, has emerged as a significant public health concern.
These opportunistic pathogens can cause a range of serious infections, including the painful and potentially blinding Acanthamoeba keratitis and the rare but often fatal granulomatous amoebic encephalitis.
Acanthamoeba species thrive in soil, water, and even air, posing a threat to both immunocompromised individuals and the general population.
Effective treatment and prevention strategies are crucial for managing Acanthamoeba-related diseases, which is why research in this area is vital.
Leveraging advanced tools and techniques, researchers can optimize their Acanthamoeba studies.
PubCompare.ai, an AI-driven platform, helps locate the most effective protocols from literature, pre-prints, and patents.
By facilitating protocol comparisons, the platform can enhance reproducibility and accuracy, taking Acanthamoeba research to new heights.
Key aspects of Acanthamoeba biology and pathogenesis include its ability to form resilient cysts, its interactions with host cells and immune responses, and its adaptations to various environmental conditions.
Researchers may employ a range of techniques, such as gentamicin exposure, AlamarBlue viability assays, and molecular methods like the QIAamp DNA Mini Kit, to study these amoebae.
The use of specialized equipment, such as the EnSpire Multimode Plate Reader, EnSpire microplate reader, and EZ Read 400 Microplate Reader, can facilitate high-throughput analyses and optimize experimental workflows.
Additionally, cell culture models, like the CellCarrier system, may be utilized to investigate Acanthamoeba-host cell interactions.
By leveraging the insights gained from the MeSH term description and the power of data-driven decision making, researchers can enhance their understanding of Acanthamoeba and develop more effective strategies for managing the associated diseases.
With a focus on reproducibility, accuracy, and innovation, the field of Acanthamoeba research continues to evolve, paving the way for improved patient outcomes.