Unusual Vermamoeba Vermiformis Strain Isolated from Snow in Mount Teide, Tenerife, Canary Islands, Spain

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

First Report of Vermamoeba vermiformis in the Island of El Hierro, Canary Islands, Spain

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 ºC. 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

Naphthyridine Derivatives Induce Programmed Cell Death in Naegleria fowleri

Primary amoebic encephalitis (PAM) caused by the opportunistic pathogen Naegleria fowleri is characterized as a rapid and lethal infection of the brain which ends in the death of the patient in more than 90% of the reported cases. This amoeba thrives in warm water bodies and causes infection after individuals perform risky activities such as splashing or diving, mostly in non-treated water bodies such as lakes and ponds. Moreover, the infection progresses very fast and no fully effective molecules have currently been found to treat PAM. In this study, naphthyridines fused with chromenes or chromenones previously synthetized by the group were tested in vitro against the trophozoite stage of two strains of N. fowleri. In addition, the most active molecule was evaluated in order to check the induction of programmed cell death (PCD) in the treated amoebae. Compound 3 showed good anti-Naegleria activity (61.45 ± 5.27 and 76.61 ± 10.84 µM, respectively) against the two different strains (ATCC® 30808 and ATCC® 30215) and a good selectivity compared to the cytotoxicity values (>300 µM). In addition, it was able to induce PCD, causing DNA condensation, damage at the cellular membrane, reduction in mitochondrial membrane potential and ATP levels, and ROS generation. Hence, naphthyridines fused with chromenes or chromenones could be potential therapeutic agents against PAM in the near future.This work was funded by PI18/01380 from the Instituto de Salud Carlos III, Spain; RICET (project RD16/0027/0001) from the Programa Redes Temáticas de Investigación Cooperativa, FIS (Ministerio Español de Salud, Madrid, Spain); and CB21/13/00100 Consorcio Centro De Investigacion Biomedica En Red M.P. (CIBER) de Enfermedades Infecciosas, Inst. de Salud Carlos III, Madrid, Spain. A.R.L. and I.A.J. were funded by the Agencia Canaria de Investigación, Innovación y Sociedad de la Información (ACIISI). Additionally, financial support from the Ministerio de Ciencia, Innovación y Universidades (MCIU), Agencia Estatal de Investigación (AEI), and Fondo Europeo de Desarrollo Regional (FEDER; RTI2018-101818-B-I00, UE) and from Gobierno Vasco, Universidad del País Vasco (GV, IT 992-16; UPV) is gratefully acknowledged. Technical and human support provided by IZO-SGI, SGIker (UPV/EHU, MICINN, GV/EJ, ERDF, and ESF) is gratefully acknowledged

Ozone Eliminates SARS-CoV-2 from Difficult-to-Clean Office Supplies and Clinical Equipment.

Background: Severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) continues to cause profound health, economic, and social problems worldwide. The management and disinfection of materials used daily in health centers and common working environments have prompted concerns about the control of coronavirus disease 2019 (COVID-19) infection risk. Ozone is a powerful oxidizing agent that has been widely used in disinfection processes for decades. The aim of this study was to assess the optimal conditions of ozone treatment for the elimination of heat-inactivated SARS-CoV-2 from office supplies (personal computer monitors, keyboards, and computer mice) and clinical equipment (continuous positive airway pressure tubes and personal protective equipment) that are difficult to clean. (2) Methods: The office supplies and clinical equipment were contaminated in an area of 1 cm2 with 1 × 104 viral units of a heat-inactivated SARS-CoV-2 strain, then treated with ozone using two different ozone devices: a specifically designed ozonation chamber (for low–medium ozone concentrations over large volumes) and a clinical ozone generator (for high ozone concentrations over small volumes). SARS-CoV-2 gene detection was carried out using quantitative real-time polymerase chain reaction (RT-qPCR). (3) Results: At high ozone concentrations over small surfaces, the ozone eliminated SARS-CoV-2 RNA in short time periods—i.e., 10 min (at 4000 ppm) or less. The optimum ozone concentration over large volumes was 90 ppm for 120 min in ambient conditions (24 °C and 60–75% relative humidity). (4) Conclusions: This study showed that the appropriate ozone concentration and exposure time eliminated heat-inactivated SARS-CoV-2 RNA from the surfaces of different widely used clinical and office supplies, decreasing their risk of transmission, and improving their reutilization. Ozone may provide an additional tool to control the spread of the COVID-19 pandemic.TRUEInstituto de Salud Carlos III, Madrid, Spain, and by the European Regional Development Funds (FEDER)Fundación Canaria del Instituto de Investigación Sanitaria de Canarias (FIISC), Las PalmasFundación Mapfre Guanarteme, Las PalmasGobierno de Canarias, Las Palmaspu

Validity and reliability of the international fItness scale (IFIS) in preschool children

Objectives Examine the validity and reliability of parent-reported International FItness Scale (IFIS) in preschoolers. Method A cross-sectional study of 3051 Spanish preschoolers (3-5 years). Fitness was measured by PREFIT battery and reported by parents using an adapted version of the IFIS. Waist circumference was evaluated, and the waist-to-height ratio (WHtR) was calculated. Seventy-six parents of randomly selected schoolchildren completed the IFIS twice for a reliability assessment. Results ANCOVA, adjusted for sex, age and WHtR, showed that preschoolers who were scored by their parents as having average-to-very good fitness had better levels of measured physical fitness than those preschoolers who were classified as having very poor/poor fitness levels (18.1laps to 22.1laps vs 15.6laps for cardiorespiratory fitness; 6.6 kg to 7.5 kg vs 5.3 kg for muscular fitness-handgrip-; 71.7 cm to 76.4 cm vs 62.0 cm for muscular fitness-standing long jump-; 17.2s to 16.2s vs 18.2s for speed/agility; and 11.2s to 15.6s vs 8.7s for balance; p < 0.001). The weighted kappa for concordance between parent-reported fitness levels and objective assessment was poor (kappa <= 0.18 for all fitness measures). Overall, the mean values of the abdominal adiposity indicators were significantly lower in high-level fitness categories reported by parents than in low-level fitness categories (p < 0.05). The test-retest reliability ranged from 0.46 to 0.62. Conclusions The reliability of the parent-reported IFIS are acceptable, but the concordance between parents reported and objectively measures fitness levels is poor, suggesting that parents' responses may not be able to correctly classify preschoolers according to their fitness level.The PREFIT project takes place thanks to the funding linked to the Ramon y Cajal grant held by Ortega FB (RYC-2011-09011)

Sesquiterpene lactones as potential therapeutic agents against Naegleria fowleri

Naegleria fowleri is the causative agent the primary amoebic meningoencephalitis (PAM), a fatal disease in more than the 90% of the reported cases that affects the central nervous system. The amoeba infects the nasal cavity of mostly children and young adults who report previous aquatic exposure in warm water sources. The rapid progression of the disease and the lack of effective and safety therapeutic options make the search of new anti-amoebic compounds an urgent issue. In this study, twelve sesquiterpene lactones isolated from the zoanthid Palythoa aff. clavata were tested against the trophozoite stage of Naegleria fowleri. Anhydroartemorin (2) and 1(10)Z,4E,14-acetoxy-costunolide (3) showed the best anti-amoeboid activity values with IC50 23.02 ± 1.26 and 28.34 ± 6.27, respectively. In addition, the mechanisms of programmed cell death induction of these two molecules were evaluated with positive results for both compounds. Finally, a structure-activity relationship was analyzed to reveal the dependence of reactivity and lipophilicity on the biological activity. The log P values of the compounds were calculated to postulate them as good candidates to cross the blood-brain barrier, a limiting factor in the development of new anti-Naegleria treatments. Therefore, the mentioned sesquiterpene lactones could be considered as potential PAM therapeutic options in the future.This work was funded by projects PI18/01380 from Instituto de Salud Carlos III, Spain and RICET (RD16/0027/0001 project) and PID2019–109476RB-C21 (BIOALGRI) (Spanish Ministry of Science, Madrid, Spain; from Programa Redes Temáticas de Investigación Cooperativa, FIS (Ministerio Español de Salud, Madrid, Spain) and FEDER. Consorcio Centro de Investigación Biomédica En Red M.P. (CIBER) de Enfermedades Infecciosas (CIBERINFEC), Instituto de Salud Carlos III, 28006 Madrid, Spain and Cabildo de Tenerife 21/0587 cofunded by MEDI and FDCAN. ARL and IAJ were funded by Agencia Canaria de Investigación, Innovación y Sociedad de la Información (ACIISI). N.N. were funded by the Agustín de Betancourt Programme (Cabildo de Tenerife, TFinnova Programme supported by MEDI and FDCAN funds). Authors acknowledge to Drs. Alberto Brito and Adriana Rodríguez Hernández from Universidad de La Laguna (ULL) the taxonomic classification of biological material, and Dr. Ezequiel Quintana Morales from Instituto de Productos Naturales y Agrobiología, Consejo Superior de Investigaciones Científicas (IPNA-CSIC) for assistance in molecular calculations.Peer reviewe

Gongolarones as antiamoeboid chemical scaffold

Free Living Amoeba (FLA) infections caused by Acanthamoeba genus include chronic nervous system diseases such as Granulomatous Amoebic Encephalitis (GAE), or a severe eye infection known as Acanthamoeba keratitis (AK). Current studies focused on therapy against these diseases are aiming to find novel compounds with amoebicidal activity and low toxicity to human tissues. Brown algae, such as Gongolaria abies-marina (previously known as Cystoseira abies-marina, S.G. Gmelin), presents bioactive molecules of interest, including some with antiprotozoal activity. In this study, six meroterpenoids were isolated and purified from the species Gongolaria abies-marina. Gongolarones A (1), B (2) and C (3) were identified as new compounds. Additionally, cystomexicone B (4), 1′-methoxyamentadione (5) and 6Z-1′-methoxyamentadione (6) were isolated. All compounds exhibited amoebicidal activity against Acanthamoeba castellanii Neff, A. polyphaga and A. griffini strains. Gongolarones A (1) and C (3) showed the lowest IC50 values against the two stages of these amoebae (trophozoite and cyst). Structure-activity relationship revealed that the cyclization by ether formation from C-12 to C-15 of 1, and the isomerization Δ2 t to Δ3 t of 3, increased the antiamoeboid activity of both compounds. Furthermore, gongolarones A (1) and C (3) triggered chromatin condensation, mitochondrial malfunction, oxidative stress, and disorganization of the tubulin-actin cytoskeleton in treated trophozoites. Moreover, transmission electron microscopy (TEM) images analysis revealed that compounds 1 and 3 induced autophagy process and inhibited the encystation process. All those results suggest that both compounds could induce programmed cell death (PCD) in Acanthamoeba.This study was supported funded by projects PID2019-109476RB-C21 (BIOALGRI) (Spanish Ministry of Science), Madrid, Spain; Fundación CajaCanarias–Fundación Bancaria “La Caixa” (2019SP52). Red de Investigación Cooperativa en Enfermedades Tropicales (RICET), Spain (project no. RD16/0027/0001 of the programe of Redes Temáticas de Investigación Cooperativa, FIS). Consorcio Centro de Investigación Biomédica en Red (CIBER), Área de Enfermedades Infecciosas (CIBERINFEC), Instituto de Salud Carlos III, 28029, Madrid, Spain (CB21/13/00100). Project No. 21/0587 funded by the Cabildo de Tenerife, Tenerife innova, Marco Estratégico de Desarrollo Insular (MEDI) and Fondo de Desarrollo de Canarias (FDCAN). Project number ProID2021010118 funded by Agencia Canaria de Investigación, Innovación y Sociedad de la Información (ACIISI). RLRE was funded by a grant from ACIISI cofunded by Fondo Social Europeo (FSE) and FEDER, (TESIS2020010117). MOM was supported by the Programa de Apoyos para la Superación del Personal Académico de la UNAM (PASPA 2021) for carrying out the research stay between the Universidad de La Laguna and la Facultad de Estudios Superiores Iztacala.Peer reviewe

First Report of a Case of Prostatitis Due to Acanthamoeba in a Dog

The first case of prostatitis in a ten year old mixed breed dog due to Acanthamoeba genotype T4 is reported. The dog was suffering from kidney dysfunction and was admitted for exploration of its organs by echography. All organs were in normal conditions with the exception of the prostate which showed signs of inflammation. An ultrasound-guided puncture was thus performed for further cytological and microbiological study. When the obtained fluid was observed under the microscope, Acanthamoeba trophozoites were detected in a high number. No other pathogens were isolated. Both culture and PCR were positive for Acanthamoeba genus and the isolate was later identified as genotype T4. Unfortunately at this stage, the dog’s owner decided to reject any kind of treatment or therapy. To the best of our knowledge, this is the first report of prostatitis in a dog due to Acanthamoeba genus