IDENTIFICATION AND CHARACTERIZATION OF PIGMENTED BACTERIA ISOLATED FROM MALAYSIAN SEAWATER

Purpose of study: Bacteria can naturally produce pigments that can be useful for various applications as they possess antimicrobial metabolites among other numerous benefits towards the human health. This study was carried out to identify the species of marine bacterial isolates PMA, PM3C1 and PM5C1 exhibiting yellow, orange and green colors respectively. Methodology: The current study is using Polymerase Chain Reaction (PCR) amplification and sequence analysis of their 16S rRNA gene. The stability of pigments extracted from the bacterial samples was also analyzed against different temperature and light conditions. Main Findings: Sequence alignment using BLAST revealed that the yellow, orange, and green-pigmented bacteria have 84% similarity with Staphylococcus aureus, 85% similarity with Exiguobacterium profundum and 95% similarity with Pseudomonas aeruginosa respectively. The green pigment showed major changes in color following exposure to sunlight and fluorescent light, and when incubated at 24°C and 50°C. Exposure to direct sunlight also results in the reduction of color for the yellow and orange extracts, while no effect was observed for both pigments under fluorescent light. Incubation at 50°C results in the reduction of the orange color, while the yellow pigment was observed to be unaffected suggesting its stability at high temperature. Implications: Natural pigments production can provide many advantages including reduction of pollution generation, ease of disposal and other benefits to the human health.

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Isolation and Characterization of Soil Myxobacteria from Nepal

Journal of Institute of Science and Technology ◽

10.3126/jist.v24i2.27246 ◽

2019 ◽

Vol 24 (2) ◽

pp. 7-16

Author(s):

Nabin Rana ◽

Saraswoti Khadka ◽

Bishnu Prasad Marasini ◽

Bishnu Joshi ◽

Pramod Poudel ◽

...

Keyword(s):

Sequence Similarity ◽

Antimicrobial Properties ◽

Antimicrobial Activities ◽

Rrna Gene ◽

Isolation And Characterization ◽

The Family ◽

Global Concern ◽

Bacteria And Fungi ◽

Antimicrobial Metabolites

Realizing myxobacteria as a potential source of antimicrobial metabolites, we pursued research to isolate myxobacteria showing antimicrobial properties. We have successfully isolated three strains (NR-1, NR-2, NR-3) using the Escherichia coli baiting technique. These isolates showed typical myxobacterial growth characteristics. Phylogenetic analysis showed that all the strains (NR-1, NR-2, NR-3) belong to the family Archangiaceae, suborder Cystobacterineae, and order Myxococcales. Furthermore, 16S rRNA gene sequence similarity searched through BLAST revealed that strain NR-1 showed the closest similarity (91.8 %) to the type strain Vitiosangium cumulatum (NR-156939), NR-2 showed (98.8 %) to the type of Cystobacter badius (NR-043940), and NR-3 showed the closest similarity (83.5 %) to the type of strain Cystobacter fuscus (KP-306730). All isolates showed better growth in 0.5-1 % NaCl and pH around 7.0, whereas no growth was observed at pH 9.0 and below 5.0. All strains showed better growth at 32° C and hydrolyzed starch, whereas casein was efficiently hydrolyzed by NR-1 and NR-2. Besides, preliminary antimicrobial tests from crude extracts showed activities against Gram-positive, Gram-negative bacteria, and fungi. Our findings suggest that the arcane soil habitats of Nepal harbor myxobacteria with the capability to produce diverse antimicrobial activities that may be explored to overcome the rapidly rising global concern about antibiotic resistance.

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Characterization of Diazotrophs Containing Mo-Independent Nitrogenases, Isolated from Diverse Natural Environments

Applied and Environmental Microbiology ◽

10.1128/aem.02694-07 ◽

2008 ◽

Vol 74 (11) ◽

pp. 3471-3480 ◽

Cited By ~ 47

Author(s):

Doris A. Betancourt ◽

Telisa M. Loveless ◽

James W. Brown ◽

Paul E. Bishop

Keyword(s):

Azotobacter Vinelandii ◽

Wood Chip ◽

Pcr Amplification ◽

Rrna Gene ◽

Natural Environments ◽

Gene Sequence Analysis ◽

Nitrogenase Genes ◽

Free Media ◽

Aerobic And Anaerobic

ABSTRACT Molybdenum-independent nitrogenases were first described in the nitrogen-fixing bacterium Azotobacter vinelandii and have since been described in other diazotrophic bacteria. Previously, we reported the isolation of seven diazotrophs with Mo-independent nitrogenases from aquatic environments. In the present study, we extend these results to include diazotrophs isolated from wood chip mulch, soil, “paraffin dirt,” and sediments from mangrove swamps. Mo-deficient, N-free media under both aerobic and anaerobic conditions were used for the isolations. A total of 26 isolates were genetically and physiologically characterized. Their phylogenetic placement was determined using 16S rRNA gene sequence analysis. Most of the isolates are members of the gamma subdivision of the class Proteobacteria and appear to be specifically related to fluorescent pseudomonads and azotobacteria. Two other isolates, AN1 and LPF4, are closely related to Enterobacter spp. and Paenibacillus spp., respectively. PCR and/or Southern hybridization were used to detect the presence of nitrogenase genes in the isolates. PCR amplification of vnfG and anfG was used to detect the genetic potential for the expression of the vanadium-containing nitrogenase and the iron-only nitrogenase in the isolates. This study demonstrates that diazotrophs with Mo-independent nitrogenases can be readily isolated from diverse natural environments.

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PROBIOTIC CHARACTERIZATION OF BACILLUS SUBTILIS STRAIN ISOLATED FROM INFANT FECAL MATTER REVEALED BY 16S rRNA GENE AND PHYLOGENETIC ANALYSIS

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2021.v14i12.43204 ◽

2021 ◽

pp. 77-85

Author(s):

DEVARANJAN DAS ◽

CHANDI CHARAN RATH ◽

NAKULANANDA MOHANTY ◽

SMITA HASINI PANDA

Keyword(s):

Antimicrobial Activity ◽

Bacillus Subtilis ◽

16S Rrna ◽

16S Rrna Gene ◽

Bile Salt ◽

Rrna Gene ◽

Probiotic Properties ◽

Fecal Matter ◽

Antimicrobial Metabolites

Objective: The rationale of our study was to isolate and identify the putative probiotic strain from infant fecal matter exhibiting a broad range of antimicrobial activity and to analyze the effect of different culturing conditions on its probiotic properties and the production of antimicrobial metabolites. Methods: In the present study, bacterial strains were screened for probiotic properties and antimicrobial activity from infant fecal matter (6 months–2 years). The effect of varying culture conditions such as tolerance to acid, bile salt, phenol, NaCl, pH, incubation period, and temperature along with autoaggregation assay, hydrophobicity, and hemolysis was studied. The characterization of the potent strain was studied by morphological, biochemical, and 16S rRNA gene sequencing along the phylogenetic affiliation of the strain was studied. Results: Two putative probiotic bacteria (DAM and IFM) were isolated, identified, characterized, and predicted at pH 2.0, 3.0, and 4.0, the isolate IFM had 50%, 60%, and 70% survivability, while isolate DAM had 55%, 63%, and 75% survivability, respectively. At a bile salt concentration of 0.5%, both isolates had a 75% survival rate. The isolates exhibited a high percentage of hydrophobicity and autoaggregation. The isolates also had non-hemolytic activity and were susceptible to many clinical tested antibiotics (tetracycline, erythromycin, ampicillin, gentamycin, penicillin, etc.). The isolate showed antimicrobial activity against enteric pathogens such as Staphylococcus aureus, Escherichia coli, and Shigella dysenteriae. The accession number of Bacillus subtilis MT279753 and MK453362 was submitted to NCBI. Conclusion: The result revealed that isolates have potent probiotic properties and possess a direct influence on the production of antimicrobial metabolites. These parameters can be modified for the improvement of the potentiality of the isolates.

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Genetic characterization of Blastocystis from wild animals in Sichuan Wolong National Natural Reserve, southwestern of China-Zoonotic potential

10.21203/rs.3.rs-471433/v2 ◽

2021 ◽

Author(s):

Shanyu Chen ◽

Wangyu Meng ◽

Ziyao Zhou ◽

Lei Deng ◽

Xiaogang Shi ◽

...

Keyword(s):

Pcr Amplification ◽

Human Infection ◽

Rrna Gene ◽

Zoonotic Potential ◽

Wild Animals ◽

Genetic Characteristics ◽

Natural Reserve ◽

Faecal Samples ◽

Wide Range

Abstract Background Blastocystis, a highly prevalent eukaryotic parasite, has been identified in a wide range of hosts, including humans, domestic and wild animals. Many animals are potential sources of Blastocystis infection for humans, while few information about the prevalence of Blastocystis in wild animals have being documented. Therefore, the present study was designed to investigate the prevalence and subtypes of Blastocystis in wild animals of Sichuan Wolong National Natural Reserve, southwestern of China, so as to assess the zoonotic potential of these animals. Methods A total of 300 faecal samples were collected from 27 wildlife species in three areas of Sichuan Wolong National Natural Reserve in southwestern China. The subtype (ST) genetic characteristics and prevalence of Blastocystis were determined by PCR amplification of the barcode region (a fragment of ∼600 bp) of the SSU rRNA gene, and phylogenetic analysis were further performed to determine the genetic characteristics of Blastocystis subtypes. Results 30 of 300 faecal samples (10.0%) were Blastocystis-positive. The highest prevalence of Blastocystis was found in Yinchanggou (18.3%), which was significantly higher than that in Niutoushan (7.5%), and Genda (5.5%) (P < 0.05). Specifically, the highest prevalence of Blastocystis was found in primates (20.0%, 1/5), followed by rodentia 14.3% (1/7), artiodactyla 13.1% (26/198), carnivora 2.3% (2/87), galliformes 0% (0/3). Sequence analysis showed 5 subtypes (ST1, ST3, ST5, ST13, and ST14), with ST13 and ST14 as the predominant subtype (33.3%, 10/30), followed by ST1 (20.0%, 6/30). Conclusions To the best of our knowledge, this is the first molecular investigation on Blastocystis infection in wild animals in southwestern of China. ST1, ST3, and ST5 were identified in both humans and wild animals, suggesting that these wild animals may be potential reservoirs of Blastocystis for human infection.

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Identification and molecular characterization of Wolbachia strains in natural populations of Aedes albopictus in China

Parasites & Vectors ◽

10.1186/s13071-020-3899-4 ◽

2020 ◽

Vol 13 (1) ◽

Cited By ~ 3

Author(s):

Yaping Hu ◽

Zhiyong Xi ◽

Xiaobo Liu ◽

Jun Wang ◽

Yuhong Guo ◽

...

Keyword(s):

Aedes Albopictus ◽

Dengue Infection ◽

Natural Populations ◽

Pcr Amplification ◽

Surface Protein ◽

Multiple Infection ◽

Rrna Gene ◽

Dengue Outbreaks ◽

Surface Protein Gene

Abstract Background Aedes albopictus is naturally infected with Wolbachia spp., maternally transmitted bacteria that influence the reproduction of hosts. However, little is known regarding the prevalence of infection, multiple infection status, and the relationship between Wolbachia density and dengue outbreaks in different regions. Here, we assessed Wolbachia infection in natural populations of Ae. albopictus in China and compared Wolbachia density between regions with similar climates, without dengue and with either imported or local dengue. Results To explore the prevalence of Wolbachia infection, Wolbachia DNA was detected in mosquito samples via PCR amplification of the 16S rRNA gene and the surface protein gene wsp. We found that 93.36% of Ae. albopictus in China were positive for Wolbachia. After sequencing gatB, coxA, hcpA, ftsZ, fbpA and wsp genes of Wolbachia strains, we identified a new sequence type (ST) of wAlbB (464/465). Phylogenetic analysis indicated that wAlbA and wAlbB strains formed a cluster with strains from other mosquitoes in a wsp-based maximum likelihood (ML) tree. However, in a ML tree based on multilocus sequence typing (MLST), wAlbB STs (464/465) did not form a cluster with Wolbachia strains from other mosquitoes. To better understand the association between Wolbachia spp. and dengue infection, the prevalence of Wolbachia in Ae. albopictus from different regions (containing local dengue cases, imported dengue cases and no dengue cases) was determined. We found that the prevalence of Wolbachia was lower in regions with only imported dengue cases. Conclusions The natural prevalence of Wolbachia infections in China was much lower than in other countries or regions. The phylogenetic relationships among Wolbachia spp. isolated from field-collected Ae. albopictus reflected the presence of dominant and stable strains. However, wAlbB (464/465) and Wolbachia strains did not form a clade with Wolbachia strains from other mosquitoes. Moreover, lower densities of Wolbachia in regions with only imported dengue cases suggest a relationship between fluctuations in Wolbachia density in field-collected Ae. albopictus and the potential for dengue invasion into these regions.

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Prevalence and Genetic Characterization of Cryptosporidium Infection in Java Sparrows (Lonchura oryzivora) in Northern China

BioMed Research International ◽

10.1155/2017/2318476 ◽

2017 ◽

Vol 2017 ◽

pp. 1-4 ◽

Cited By ~ 2

Author(s):

Qiu-Xia Yao ◽

Xiao-Xuan Zhang ◽

Kai Chen ◽

Jian-Gang Ma ◽

Wen-Bin Zheng ◽

...

Keyword(s):

Pcr Amplification ◽

Northern China ◽

Small Subunit ◽

Rrna Gene ◽

Ssu Rrna ◽

Ssu Rrna Gene ◽

Wide Range ◽

Baseline Information ◽

And Control

Cryptosporidiosis is a cosmopolitan parasitosis that affects a wide range of hosts including birds. As information concerning Cryptosporidium in birds is limited, the present study examined the prevalence and genotypes of Cryptosporidium in Java sparrows in Beijing and Shangqiu, northern China. Three hundred and fifty fecal samples were collected from Java sparrows (Lonchura oryzivora, 225 white Java sparrows and 125 gray Java sparrows) in Beijing and Shangqiu in October 2015, and the samples were examined by PCR amplification of the small subunit ribosomal RNA (SSU rRNA) gene. The overall Cryptosporidium prevalence is 13.42% (47/350), with 16.44% (37/225) in white Java sparrows and 8.00% (10/125) in gray Java sparrows. Cryptosporidium prevalence was 9.82% (16/163) in Java sparrows from Beijing and 16.58% (31/187) in Java sparrows from Shangqiu. The prevalence of Cryptosporidium in females and males was 40.63% (26/64) and 7.34% (21/286), respectively. The Cryptosporidium prevalence in Java sparrows of different ages varied from 10.47% to 16.33%. Sequence analysis of the SSU rRNA gene revealed that all the samples represented C. baileyi. This is the first report of Cryptosporidium in gray Java sparrows in China, which extend the host range for C. baileyi. These results provide baseline information for further studies of molecular epidemiology and control of Cryptosporidium infection in poultry in China.

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Histopathology and Molecular Characterization of Microsporidian Parasites in Cultured Shrimp Penaeus Monodon Fabricius 1798 of Bangladesh

Journal of the Asiatic Society of Bangladesh Science ◽

10.3329/jasbs.v45i2.46592 ◽

2019 ◽

Vol 45 (2) ◽

pp. 187-196 ◽

Cited By ~ 1

Author(s):

Md Munjur Hossain ◽

Nusrat Jahan Punom ◽

Md Mostavi Enan Eshik ◽

Mst Khadiza Begum ◽

Md Aminul Islam Bhuiyan ◽

...

Keyword(s):

Penaeus Monodon ◽

Pcr Amplification ◽

Histological Section ◽

Rrna Gene ◽

Microsporidian Parasite ◽

Specific Primers ◽

West Region ◽

Field Samples ◽

Enterocytozoon Hepatopenaei

Black tiger shrimp (Penaeus monodon Fabricius 1798) cultured in Bangladesh was investigated for the presence of microsporidian parasite. Histological section of hepatopancreas showed a large number of microsporidian spores under light microscopy. Spores under scanning electron microscope appeared oval shapes. Histology of infected shrimps showed severe degeneration of hepatopancreatic tubules. Early and late stage of microsporidian parasites in hepatopancreatic tubules were also observed. DNA extracted from the hepatopancreas of shrimps were subjected to PCR amplification using primers targeting microsporidian SSU rRNA gene. The PCR amplified an expected product of ~328 bp and the sequences showed 81 - 82% identity with the Paranucleospora theridion reported from western Norway in 2008. Further screening of field samples was carried out using EHP-specific primers. DNA extracted from ten hepatopancreas samples of P. monodon were tested and none found to be positive for EHP (Enterocytozoon hepatopenaei). This is the first report for the identification of microsporidian parasites in cultured shrimp along the south-west region of Bangladesh. Asiat. Soc. Bangladesh, Sci. 45(2): 187-196, December 2019

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Comparative genome characterization of Echinicola marina sp. nov., isolated from deep-sea sediment provide insight into carotenoid biosynthetic gene cluster evolution

Scientific Reports ◽

10.1038/s41598-021-03683-0 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Yu Pang ◽

Mengru Chen ◽

Wei Lu ◽

Ming Chen ◽

Yongliang Yan ◽

...

Keyword(s):

Deep Sea ◽

Genomic Analysis ◽

Biosynthetic Gene Cluster ◽

Rrna Gene ◽

Deep Sea Sediment ◽

Cluster Evolution ◽

Gene Sequence Analysis ◽

Nacl Concentration ◽

Pigmented Bacteria

AbstractEchinicola, carotenoid-pigmented bacteria, are isolated from various hypersaline environments. Carotenoid accumulation in response to salt stress can stabilize the cell membrane in order to survive. A pink-colored strain SCS 3–6 was isolated from the deep-sea sediment of the South China Sea. Growth was found to occur at 10–45 °C. The strain could tolerate 10% (w/v) NaCl concentration and grow at pH 5–9. The complete genome of SCS 3–6 comprises 5053 putative genes with a total 5,693,670 bp and an average G + C content of 40.11 mol%. The 16S rRNA gene sequence analysis indicated that strain SCS 3–6 was affiliated with the genus Echinicola, with the closely strains were Echinicola arenosa CAU 1574T (98.29%)and Echinicola shivajiensis AK12T (97.98%). For Echinicola species with available genome sequences, pairwise comparisons for average nucleotide identity (ANI) and in silico DNA-DNA hybridization (DDH) revealed ANIb values from 70.77 to 74.71%, ANIm values from 82.72 to 88.88%, and DDH values from 18.00 to 23.40%. To identify their genomic features, we compared their genomes with those of other Echinicola species. Phylogenetic analysis showed that strain SCS 3–6 formed a monophyletic clade. Genomic analysis revealed that strain SCS 3–6 possessed a complete synthetic pathway of carotenoid and speculated that the production was astaxanthin. Based on phenotypic and genotypic analyses in this study, strain SCS 3–6 is considered to represent a novel species of the genus Echinicola for which the name Echinicola marina sp. nov. is proposed. The type strain is SCS 3-6T (= GDMCC 1.2220T = JCM 34403T).

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Molecular and biochemical characterization of Pseudomonas putida isolated from bottled uncarbonated mineral drinking water

Archives of Biological Sciences ◽

10.2298/abs1401023t ◽

2014 ◽

Vol 66 (1) ◽

pp. 23-28 ◽

Cited By ~ 1

Author(s):

S. Tasic ◽

M. Kojic ◽

D. Obradovic ◽

Irena Tasic

Keyword(s):

Drinking Water ◽

16S Rrna ◽

Pseudomonas Putida ◽

Molecular Characterization ◽

Biochemical Characterization ◽

Pcr Amplification ◽

Identification System ◽

Rrna Gene ◽

Identification Software

Pseudomonas putida belongs to a group of opportunistic pathogens that can cause disease in people with weakened or damaged immune systems. Some strains have medical significance, and for most ingestion is not the primary route of infection. If water used by predisposed subjects is contaminated by P. putida, they may become ill. The aim of this work was the biochemical and molecular characterization of strain ST3 of P. putida isolated from non-carbonated bottled drinking water from Jakov Do 4 on Mt. Vlasina. Characterization of P. putida was performed to assess the risk to human health of the indigenous strains present in the water. Biochemical characterization of strains was performed using the manual identification system ID 32 GN (BioM?rieux). Identification was obtained using the database identification software ATB System (Bio-M?rieux). Molecular characterization was performed by PCR amplification and 16S rDNA ?thermal cycling sequencing?. Biochemical identification of the strain ST3 was accurate (Id = 99.8%). Comparing the sequences obtained for strain ST3 with NCBI gene bank sequences for 16S rRNA, the highest similarity of our strain (96% identity) with a strain of P. putida, designated as biotype A (gi|18076625|emb|AJ308311.1|.PPU308311) isolated in New Zealand, was obtained. While comparison with the NCBI collection of all deposited sequences showed that the 16S rRNA gene sequence of strain ST3 has very high homology, it is not identical, indicating indirectly that strain ST3 is an indigenous strain. This article has been corrected. Link to the correction <a href="http://dx.doi.org/10.2298/ABS151023125E">10.2298/ABS151023125E</a>

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Genetic characterization of Blastocystis from wild animals in Sichuan Wolong National Natural Reserve, southwestern of China-Zoonotic potential

10.21203/rs.3.rs-471433/v1 ◽

2021 ◽

Author(s):

Shanyu Chen ◽

Wangyu Meng ◽

Ziyao Zhou ◽

Lei Deng ◽

Xiaogang Shi ◽

...

Keyword(s):

Pcr Amplification ◽

Human Infection ◽

Rrna Gene ◽

Zoonotic Potential ◽

Wild Animals ◽

Genetic Characteristics ◽

Natural Reserve ◽

Faecal Samples ◽

Wide Range

Abstract Background: Blastocystis, a highly prevalent eukaryotic parasite, has been identified in a wide range of hosts, including humans, domestic and wild animals. Many animals are potential sources of Blastocystis infection for humans, while few information about the prevalence of Blastocystis in wild animals have being documented. Therefore, the present study was designed to investigate the prevalence and subtypes of Blastocystis in wild animals of Sichuan Wolong National Natural Reserve, southwestern of China, so as to assess the zoonotic potential of these animals.Methods: A total of 300 faecal samples were collected from 27 wildlife species in three areas of Sichuan Wolong National Natural Reserve in southwestern China. The subtype (ST) genetic characteristics and prevalence of Blastocystis were determined by PCR amplification of the barcode region (a fragment of ∼600 bp) of the SSU rRNA gene, and phylogenetic analysis were further performed to determine the genetic characteristics of Blastocystis subtypes.Results: 30 of 300 faecal samples (10.0%) were Blastocystis-positive. The highest prevalence of Blastocystis was found in Yinchanggou (18.3%), which was significantly higher than that in Niutoushan (7.5%), and Genda (5.5%) (P < 0.05). Specifically, the highest prevalence of Blastocystis was found in primates (20.0%, 1/5), followed by rodentia 14.3% (1/7), artiodactyla 13.1% (26/198), carnivora 2.3% (2/87), galliformes 0% (0/3). Sequence analysis showed 5 subtypes (ST1, ST3, ST5, ST13, and ST14), with ST13 and ST14 as the predominant subtype (33.3%, 10/30), followed by ST1 (20.0%, 6/30).Conclusions: To the best of our knowledge, this is the first molecular investigation on Blastocystis infection in wild animals in southwestern of China. ST1, ST3, and ST5 were identified in both humans and wild animals, suggesting that these wild animals may be potential reservoirs of Blastocystis for human infection.

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