rDNA Gene Sequence Research Articles

Biodegradation of Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by Newly Isolated Penicillium oxalicum SS2 in Soil Microcosms and Partial Characterization of Extracellular Depolymerase.

A fungus, designated as strain SS2 able to degrade aliphatic polyesters, poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), was isolated from soil. Strain SS2 was identified through rDNA gene sequencing and showed maximum closeness to Penicillium oxalicum. The newly isolated P. oxalicum strain SS2 had completely degraded PHB and PHBV both in emulsion and films form within 36-48h at 30°C. Furthermore, P. oxalicum SS2 degraded PHB and PHBV films in soil environment in lab-built soil microcosms within 1 week. The polymer films were evaluated for changes after degradation through scanning electron microscopy (SEM), nuclear magnetic resonance (NMR), differential scanning calorimetry (DSC) and Fourier Transform Infrared spectroscopy (FTIR). The PHBV depolymerase enzyme was purified to homogeneity through column chromatography and molecular mass was found approximately 36kDa. The depolymerase was stable over a wide range of temperature (15-60°C) and pH (3.0-8.0) with optimum 40°C and pH 5.0. The enzyme activity was significantly affected by various metal ions and surfactants. The enzyme activity was strongly enhanced in the presence of divalent cationic metal Cu2+ while inhibited by Zn2+ and non-polar detergents Tween 20 and Tween 60. Finally, it is concluded that P. oxalicum strain SS2 has profound degradation capabilities, and can be applied for the treatment of plastic-contaminated environments.

Short Communication: Sarocladium oryzae associated with sheath rot disease of rice in Indonesia

Abstract. Pramunadipta S, Widiastuti A, Wibowo A, Suga H, Priyatmojo A. 2020. Short Communication: Sarocladium oryzae associated with sheath rot disease of rice in Indonesia. Biodiversitas 21: 1243-1249. One of the obstacles in increasing rice production is the presence of sheath rot pathogen infection, which causes changes in color on the rice sheath to brown or reddish-brown, sometimes does not produce rice grain. The major fungal pathogens that cause sheath rot disease are Sarocladium oryzae and Fusarium spp. The loss of rice yields reaches 85%. The disease found in six provinces, some of which are the largest rice-producing centers in Indonesia. A total of twenty-four Sarocladium sp. were isolated from leaf sheath symptom on potato dextrose agar and water agar medium. Sheath rot pathogen identification based on molecular method was performed using internal transcribed spacer (ITS) rDNA gene sequencing. Necrosis occurs after artificial inoculation in Ciherang rice variety was observed and showed that all isolates were pathogenic. Morphological characterization of the isolates identified them as Sarocladium sp. Molecular identification showed that six representatives isolates belonging to S. oryzae. These findings are important information about the fungal pathogen that causes sheath rot disease in Indonesia, and in studies for formulating control measures of the pathogen in the future to prevent the disease epidemic on rice. This is the first report about the existence of sheath rot disease, morphological characterization and molecular identification of S. oryzae in various rice fields in Indonesia.

Bioremediation of arsenic by soil methylating fungi: Role of Humicola sp. strain 2WS1 in amelioration of arsenic phytotoxicity in Bacopa monnieri L.

Fungi mediated arsenic (As) stress modulation has emerged as an important strategy for the mitigation of As mediated stress management in plants for reducing As contamination to the food chain. In the present study, total of 45 fungal strains were isolated from the three As contaminated sites of West Bengal, India. These strains were morphologically different and inhibited variable As tolerance (10 to 5000mgl-1As). Total 21 fungal isolates, tolerant up to 5000mgl-1 AsV, were investigated for As removal (10mgl-1 As) after 21d of cultivation under laboratory conditions. The As bioaccumulation in fungal biomass ranged between 0.146 to 11.36gkg-1 biomass. Range of volatilized As was between 0.05 to 53.39mgkg-1 biomass. Most promising bioaccumulation and biovolatilization potential were observed in strains viz., 2WS1, 3WS1 and 2WS9. Strain 2WS1 showed highest As biovolatilization (53.39mgkg-1 biomass) and was identified as Humicola sp. using ITS/5.8S rDNA gene sequencing. This is the first report of Humicola sp. having As biomethylation property. Best first 8 As biomethylating fungal strains were further tested for their As remediation and PGP potential in Bacopa monnieri plant grown in As contaminated soil (20mgkg-1) in a pot experiment under greenhouse conditions. The highest leaf stem ratio and lowest As content in leaf tissues were observed in 2WS1 inoculated Bacopa monnieri plants. The presence of arsM gene in 2WS1 strain suggests As biovolatilization as possible bioremediation and As stress mitigation strategy of 2WS1. Therefore, application of this strain of Humicola sp. strain 2WS1 in As contaminated soils could be a potential and realistic mitigation strategy for reducing As contamination to cropping system coupled with enhanced productivity.

An assessment of the aggregation and probiotic characteristics of Lactobacillus species isolated from native (desi) chicken gut

SUMMARY The microflora within a chicken's gastrointestinal tract (GIT) has extensive bacterial diversity and may be a good source for the isolation of effective probiotic candidates. The resistance of desi chickens to various biotic and abiotic stresses is related to the presence of lactic acid bacteria (LAB) in their digestive system. In this study, LAB were isolated and cultured from the caecum contents of desi chickens and identified based on their phenotypic characters and rDNA gene sequences. The aggregation capability of the isolated Lactobacillus species was tested and those strains showing strong aggregation were further tested to see if they were better able to withstand the stress of GIT digestive conditions. The results demonstrated significant variation in aggregation ability and GIT stress tolerance. The autoaggregation potential of the strains was not correlated with their cell surface hydrophobicity but was found to be linked to coaggregation ability with pathogens. No potentially harmful enzymatic activities were observed in the poultry-derived LAB strains and the ones with good aggregation potential were susceptible to most common antibiotics. This study proved that native chickens can be a good source for isolating potentially effective probiotic LAB species. Furthermore, the aggregation potential among Lactobacillus species was highly strain specific and did not correlate with their ability to survive the stresses of digestion in the GIT.

Characterization of Cellobiohydrolases from Schizophyllum commune KMJ820.

A novel cellobiohydrolase (CBH)-generating fungi have been isolated and categorized as Schizophyllum commune KMJ820 based on morphology and rDNA gene sequence. Cellulose powder was used as carbon source, the total enzyme activity was 11.51U/ml is noted; which is among the highest amounts of CBH-generating microbes studied. CBH have been purified to homogenize, with pursual of serial chromatography using S. commune supernatants and two different CBHs were found; CBH 1 and 2. The filtered CBHs showed greater activity (V max = 51.4 and 20.8U/mg) in contrast to CBHs from earlier studies. The MW (molecular weights) of S. commune CBH 1 and 2 were verified to be approximately 50kDa and 150kDa, respectively, by size exclusion chromatography. Even though CBHs have been evaluated from other sources, but S. commune CBH is prominent in comparison to other CBHs by its high enzyme activity.

Isolation and identification of the spoilage microorganisms in Sichuan homemade Paocai and their impact on quality and safety.

The spoilage microbiology in Paocai (fermented vegetables) affects not only the quality of this popular traditional Chinese food but also its safety. This study aimed to isolate and identify the microorganisms commonly leading to spoilage in homemade Paocai from the Sichuan region and, further, to scientifically assess the impact of these microorganisms on product quality and safety. Seven putrid Paocai samples were collected from 7 families in different Sichuan cities. From these, 45 strains were isolated by means of a nutrient agar medium and rose bengal agar. All of the 22 strains (5 fungi and 17 bacteria) with different colonial morphologies and corruption phenomenon in Paocai were determined 16S rDNA/18S rDNA gene sequences. Bacteria were identified as Bacillus spp. (16 strains) and Staphylococcus saprophyticus (1 strain), while the 5 fungi were identified as Candida spp. (3 strains), Kodamaea ohmeri (1 strain), and Geotrichum candidum (1 strain). Based on the results of identification, 7 representative strains of different species were determined as the spoilage characteristics in paocai. All the representative strains metabolized to produce nitrite. Strain SPF21 (K. ohmeri) has a particularly serious impact on the crunchiness of Paocai; however, strains SPF19 (Bacillus subtilis) and SPF21 have the greatest influence on its color. All five of the fungi were seen to form a film on the surface of the Paocai, with SPF5 (G. candidum) exerting the most extreme influence. The growth characteristics in the broth showed that all the representative strains investigated metabolized most of the carbohydrates and were able to tolerate the salinity and acidity of the ordinary homemade Paocai. Moreover, these strains were found to have obvious impacts on the volatile components of Paocai, especially SPF2 and SPF8, with higher dimethyl disulfide and dimethyl trisulfide, which were found to have a pungent odor when highly concentrated.

Screening and identification of Aspergillus activity against Xanthomonas oryzae pv. oryzae and analysis of antimicrobial components.

To screen for Aspergillus activity against Xanthomonas oryzae pv. oryzae and analyse the antimicrobial components involved, 60 Aspergillus spp. were isolated and purified from fruits, soil and other habitats. As-75, an Aspergillus strain that can antagonize Xanthomonas oryzae pv. oryzae, was identified based on the zone of inhibition formed during co-culture. According to morphological, ITS rDNA gene sequencing and phylogenetic tree results, the strain showed close homology to Aspergillus sclerotiorum. The biochemical characterization tests showed that the fermentation broth of strain As-75 exhibited a high capacity for environmental adaptation. The results of the antimicrobial spectrum experiments demonstrated that As-75 exhibited fairly strong antagonistic activity against five plant pathogenic fungi and six plant pathogenic bacteria in vitro. The fermentation broth of strain As-75 displayed maximum stability under fluorescent illumination at temperatures below 60°C at pH 6.5. A substance with antagonistic activity was obtained from strain As-75 via fractional extraction, silica gel column chromatography and thin-layer chromatography. Through mass spectrometry, nuclear magnetic resonance and electrospray ionization mass spectrometry (ESI-MS) analyses, the target compound was identified as (2Z)-2-butenedioic acid-2-(1-methylethenyl)-4-methyl ester; its molecular weight of 170.06 daltons and formula of C8H10O4 identify it as a novel compound. Trials of the preventative and curative effects demonstrated that compound S1 exhibited a better control efficiency than the control against rice bacterial blight. Additionally, the M1 processing method was better, and the efficiency of compound S1 in preventing rice bacterial blight in six rice varieties, TN1, IR24, ZF802, Zhonghua 11, Wuyunjing 21, and Nipponbare, was 78.3%, 77.5%, 74.2%, 75.3%, 70.9%, and 72.1%, respectively.

First report of 16SrII-D phytoplasma associated with eggplant phyllody in China

In April 2018, symptoms of phyllody, witches’ broom and little leaf were observed on eggplant on a farm located in Huiyang district, Guangdong province, China. Total DNA was extracted from the leaves of symptomatic plants and subjected to molecular detection using the phytoplasma universal primers R16mF2/mR1, P1/P7 and SecAfor1/SecArev3, which showed PCR products of the expected 1.4 kb, 1.8 kb and 0.8 kb fragments, respectively. The 16S rDNA sequence showed the highest similarity (100%) with ‘Celosia argentea’ phytoplasma strain Ca1 (GenBank accession KX426374), Crotalaria witches’ broom phytoplasma strain CrWB-Hnsy1 (GenBank accession EU650181) and pear decline phytoplasma (Taiwan II) isolate Heping (GenBank accession EF193157) from the 16SrII group. Phylogenetic analyses based on 16S rDNA and secA gene sequences confirmed the closer relationship of the eggplant phyllody phytoplasma (EPP) strain with the 16SrII-D subgroup. Virtual restriction fragment length polymorphism (RFLP) analysis of the 16S rDNA sequence indicated the eggplant phyllody phytoplasma belongs to subgroup D within the 16SrII group (16SrII-D). This is the first report of eggplant phyllody phytoplasma in China.

Assessment of Plant Growth Promoting and Abiotic Stress Tolerance Properties of Wheat Endophytic Fungi.

The aims of the present work were to isolate and characterize fungal endophytic communities associated with healthy wheat (Triticum aestivum L.) plants, collected from the North China. Segregated endophytes were screened for their PGP traits, abiotic stresses (heavy metals, salinity, drought, and temperature), and antibiotic sensitivity. A total of 16 endophytic fungi were isolated using the culture-dependent approach from different tissue parts of wheat plants. Based upon their internal transcribed spacer (ITS) rDNA gene sequencing, 15 out of 16 isolates were selected for further analysis. In the contemporary investigation, a number of the tested endophytes exhibited fairly good 1-aminocyclopropane-1-carboxylic acid deaminase (ACCD) (0.03±0.011 to 1.43±0.01 µmol α-KB mg−1 protein hr−1), indole acetic acid (IAA) (1.125±0.04 to36.12±0.004µgml−1), and phosphate solubilizing index (PSI) (2.08±0.03to5.16±0.36) activities. More than 30% isolates gave positive result for siderophore and ammonia tests, whereas all exhibited catalase activity but only 2 (582PDA1 and 582PDA11) produced hydrogen cyanide. Trichoderma strains showed salt, heavy metals, and drought tolerance at high levels and also exhibited resistance to all the tested antibiotics. Strain 582PDA4 was found to be the most temperature (55°C) tolerant isolate. The findings of this study indicated that the microbial endophytes isolated from wheat plants possessing a crucial function to improve plant growth could be utilized as biofertilizers or bioagents to establish a sustainable crop production system.

A new record of Trichocladium griseum in Korea: morphological and molecular characterization

A unrecorded species of Trichocladium, Trichocladium griseum, was isolated in 2017 during a survey of fungal diversity in Ulsan province, South Korea. This species was identified based on morphological characteristics and phylogenetic analysis of the internal transcribed spacer (ITS) rDNA and β-tubulin gene sequences. T. griseum has not yet been reported in South Korea. Thus, we report for the first time a new record of Trichocladium griseum in Korea, and we include the descriptions and morphological illustrations of this fungus.

Biodegradation of 17\u03b2-estradiol by Bacterial Co-culture Isolated from Manure

Animal wastes are potential sources of natural and steroidal estrogen hormones into the environment. These hormones can be removed by microorganisms with induced enzymes. Two strains of 17β-estradiol-degrading bacteria (LM1 and LY1) were isolated from animal wastes. Based on biochemical characteristics and 16 S rDNA gene sequences, we identified strains LM1 and LY1 as belonging to the genus of Acinetobacter and Pseudomonas, respectively. Bacterial co-culture containing LM1 and LY1 bacterial strains could rapidly remove approximately 98% of E2 (5 mg L−1) within 7 days. However, strains LM1 and LY1 degraded 77% and 68% of E2 when they were incubated alone, respectively. More than 90% of 17β-estradiol (E2, ≤ 20 mg L−1) could be removed by bacterial co-culture. Low C/N ratio (1:35) was more suitable for bacterial growth and E2 degradation. The optimal pH for bacterial co-culture to degrade E2 ranged from 7.00 to 9.00. Coexisting sodium acetate, glucose and sodium citrate decreased E2 degradation in the first 4 days, but more E2 was removed when they were depleted. The growth of the bacterial co-culture was not significantly decreased by Ni, Pb, Cd or Cu at or below 0.8, 1.2, 1.6 or 0.8 mg L−1, respectively. These data highlight the usefulness of bacterial co-culture in the bioremediation of estrogen-contaminated environments.

Optimization of chromium and tannic acid bioremediation by Aspergillus niveus using Plackett\u2013Burman design and response surface methodology

A chromium and tannic acid resistance fungal strain was isolated from tannery effluent, and identified as Aspergillus niveus MCC 1318 based on its rDNA gene sequence. The MIC (minimum inhibitory concentration) of the isolate against chromium and tannic acid was found to be 200 ppm and 5% respectively. Optimization of physiochemical parameters for biosorption of chromium and tannic acid degradation was carried out by Plackett–Burman design followed by response surface methodology (RSM). The maximum chromium removal and tannic acid degradation was found to be 92 and 68% respectively by A. niveus. Chromium removal and tannic acid degradation was increased up to 11 and 6% respectively after optimization. Scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) was used to investigate biosorption phenomena.

Biogenic amines content and assessment of bacterial and fungal diversity in stinky tofu – A traditional fermented soy curd

Microbial profiles of eighteen soy curd (stinky tofu) samples as well as their brines, collected from six different regions, were assessed. In addition, biogenic amines were also quantified using HPLC to determine their levels in Chinese fermented soy curd. A total of 444 bacteria, comprising 80 species, and 116 fungal isolates, comprising 26 species, were identified using 16 S rDNA gene and internal transcribed spacer (ITS) rDNA gene sequencing, respectively. In all samples, the average number of bacteria in tofu and brine was 7.00 log cfu/g and 6.99 log cfu/mL, respectively, while the average value of fungal cells in tofu and brine were 2.58 log cfu/g and 3.55 log cfu/mL, respectively. The results revealed the predominance of Streptococcus lutetiensis, Lactococcus lactis and Yarrowia lipolytica in the samples tested. In case of biogenic amines, tyramine was detected in all samples while spermine could not be detected. β-phenethylamine was only detected in the tofu of Xi'an. The microbial composition and biogenic amines content are the main factors affecting the safety of stinky tofu products. This study indicates that a large microbial diversity is associated with the fermentation of stinky tofu which may have a key role in the production of biogenic amines.

Mycobacterium goodii endocarditis following mitral valve ring annuloplasty

BackgroundMycobacterium goodii is an infrequent human pathogen which has been implicated in prosthesis related infections and penetrating injuries. It is often initially misidentified as a gram-positive rod by clinical microbiologic laboratories and should be considered in the differential diagnosis.Case presentationWe describe here the second reported case of M. goodii endocarditis. Species level identification was performed by 16S rDNA (ribosomal deoxyribonucleic acid) gene sequencing. The patient was successfully treated with mitral valve replacement and a prolonged combination of ciprofloxacin and trimethoprim/sulfamethoxazole.ConclusionConfirmation of the diagnosis utilizing molecular techniques and drug susceptibility testing allowed for successful treatment of this prosthetic infection.

APPLE PROLIFERATION DISEASE IN CROATIAN ORCHARDS: A MOLECULAR CHARACTERIZATION OF ‘CANDIDATUS PHYTOPLASMA MALI’

Apple proliferation (AP) causes major economic losses to the European apple industry. The causal agent associated with AP is ‘Candidatus Phytoplasma mali’ which belongs to the same phylogenetic cluster as two other fruit tree phytoplasmas: ‘Ca. P. pyri’ and ‘Ca. P. prunorum’. ‘Ca. P. pyri’ and ‘Ca. P. solani’ have sporadically been associated with symptomatic apples in Croatia in previous surveys. There was no molecular evidence of ‘Ca. P. mali’ presence until the start of this study, when it was confirmed for the first time. In this four-year survey (2011-2014), the detection and characterization of ‘Ca. P. mali’ from apples and psyllid vectors was carried out. Detection was based on PCR-RFLP analysis of the 16S rRNA gene. Real-time PCR was also used for amplifying ‘Ca. P. mali’ 16S rDNA and Malus domestica chloroplast tRNA leucine gene. Phylogenetic analysis of 16S rDNA and aceF gene sequences confirmed the identification of selected phytoplasma strains. Considerable aceF variability among the strains was detected, and a new, yet unrecorded aceF genotype was found. ‘Ca. P. mali’ has consistently been identified with high incidence in apple samples from Croatian continental region orchards, whereas it occurred sporadically in the coastal areas. This is the first molecular detection and characterization of ‘Ca. P. mali’ strains from both apple (Malus domestica Burkh.) and psyllid vector Cacopsylla picta (Forster) in Croatia.

Optimization of Process Conditions for the Production of Holocellulase by a <i>Bacillus</i> Species Isolated from Nahoon Beach Sediments

Production of holocellulase by a newly isolated marine Bacillus species via submerged fermentation technique at varying cultivation conditions was investigated. Enzyme production was optimized by altering one variable per time. Moreover effect of lignocellulosic saccharides in inducing cellulase and xylanase production was also investigated. The 16S ribosomal Deoxyribonucleic Acid (16S rDNA) gene sequence analysis exhibited 98% sequence similarity of the isolate with other Bacillus species in the gene bank and was deposited with the accession number KX524510. Optimal xylanase and cellulase production was attained at pH 7, temperature 30°C and agitation speed of 50 and 150 rpm. Furthermore, maximum xylanase and cellulase production were both achieved at 60 h corresponding to the late stationary growth stage, with activity of 16.6 and 0.061 U mL-1 respectively. Xylanase production was maximally induced by beechwood xylan, xylose and arabinose with activities of 13.59, 8.78 and 1.90 (U mL-1) respectively; while cellulase production was induced by carboxymethyl cellulose only and no cellulase activity was detected in the culture supernatant of the other carbon sources tested. Optimization increased cellulase and xylanase yields being 0.006-0.061 and 0.23-16.6 (U mL-1) from unoptimized to optimized respectively. The results of the study suggest the bacterial strain to be a proficient producer of cellulase and xylanase with potentials in biotechnological application.

Characterization and treatment of colored textile wastewaters from the typical Tunisian hat Chechia dyeing using newly isolated Aspergillus niger

This study aimed to characterize and investigate, for the first time, the treatment of real colored wastewaters from the artisanal dyeing of the typical Tunisian hat Chechia, using a newly isolated fungal strain. This textile effluent was a mixture called Mix of colored wastewaters from the three main types of C hechia . The major pollutant of the Mix was the toxic Azo dye Amaranth Acid or Acid Red 27. The fungal strain that made the cleanup was discovered in a C hechia dyeing wastewater’s container and identified by ITS rDNA gene sequencing. This isolated Aspergillus niger showed interesting performances on the demonstration of Chechia wastewater’s biodegradation in batch cultures. In order to understand the effect of agitation, Mix dilution and inoculum size on decolourisation and pollution removal, a full factorial experimental design 2 3 was set up. At the optimal conditions which were 20% inoculum size, 25% Chechia Mix dilution and an agitation of 100 rpm, Aspergillus niger was able to remove color as high as 70.18±2.84% at an initial dye concentration of 1346.6±0.01 mg/L, and to reduce COD to 74.17±14.52% at an initial COD of 4157±422 mg/L. FT-IR spectra analysis confirmed the decolourisation by biodegradation and transformation of the dyes. The treatment by the isolated Aspergillus niger could be successfully applied as a sustainable method to solve one of handicraft dyeing plants environmental management issues.

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A novel organic solvent- and detergent-stable serine alkaline protease from Trametes cingulata strain CTM10101

A protease-producing fungus was isolated from an alkaline wastewater of chemical industries and identified as Trametes cingulata strain CTM10101 on the basis of the ITS rDNA gene-sequencing. It was observed that the fungus strongly produce extracellular protease grown at 30°C in potato-dextrose-broth (PDB) optimized media (13500U/ml). The pure serine protease isolated by Trametes cingulata (designated SPTC) was purified by ammonium sulfate precipitation-dialysis followed by heat-treatment and UNO S-1 FPLC cation-exchange chromatography. The chemical characterization carried on include phisico-chemical determination and spectroscopie analysis. The MALDI-TOF/MS analysis revealed that the purified enzyme was a monomer with a molecular mass of 31405.16-Da. The enzyme had an NH2-terminal sequence of ALTTQTEAPWALGTVSHKGQAST, thus sharing high homology with those of fungal-proteases. The optimum pH and temperature values of its proteolytic activity were pH 9 and 60°C, respectively, and its half-life times at 60 and 70°C were 9 and 5-h, respectively. It was completely inhibited by PMSF and DFP, which strongly suggested its belonging to the serine protease family. Compared to Flavourzyme®500L from Aspergillus oryzae and Thermolysin typeX from Geobacillus stearothermophilus, SPTC displayed higher levels of hydrolysis, substrate specificity, and catalytic efficiency as well as elevated organic solvent tolerance and considerable detergent stability. Finally, SPTC could potentially be used in peptide synthesis and detergent formulations.

Isolation and Molecular Identification of Vermamoeba vermiformis Strains from Soil Sources in El Hierro Island, Canary Islands, Spain

Free-living amoebae (FLA) are widely distributed protozoa in the environment and have been isolated from many sources such as dust, soil and water. Furthermore, some genera/species of FLA such as Naegleria fowleri, Balamuthia mandrillaris and Acanthamoeba spp. are also able to cause opportunistic infections in humans and other animals. More recently, FLA have been reported to be environmental carriers of pathogenic bacteria, fungi and viruses, and thus have gained further importance from the public health point of view. Among them, Acanthamoeba spp. and Vermamoeba vermiformis have been described in many occasions as the most common carriers of pathogens of high medical relevance such as Legionella pneumophila and Mycobacterium spp. In this study, 24 soil samples were collected from the island of El Hierro, Canary Islands, Spain, in order to check for the presence of V. vermiformis strains in these samples. Soil samples were cultured on 2% non-nutrient agar plates covered with a thin layer of heat-killed E. coli and checked daily for the presence of Vermamoeba. After a week, V. vermiformis amoebae were observed in 5 of the 24 processed samples (20.8%) incubated at room temperature and 37°C. Molecular characterization was carried out by amplifying the 18S rDNA gene and DNA sequencing, confirming that the isolated strains belonged to Vermamoeba vermiformis species. The high percentage of V. vermiformis in the studied soil sources should raise awareness in the region since these amoebae are potential environmental carriers of pathogens of high medical relevance.