Considering the limit of species identified to this date, there is the potential that this microbe could be a new species. Based on multiple 1a). Abstract. They may be domed, shining, mucoid-type colonies with radial striations or may appear smooth with entire edges. Planococcus citreus is Gram stain positive and aerobic. The isolate failed to grow on EMB or MAC plates, considering that both plates are selective for gram negative, these results confirm the results of the gram positive test: Image 4. Head The negative results and lack of growth in the API 20e, MAC, and EMB are all consistent with the microbe’s gram-positive cell wall. Considering that the API 20e test is designed for enteric gram negative microbes these results are confirmation that the microbe is gram-positive. Planococcus sp. In general, the colonies appear as white, smooth colonies. The susceptibility of the isolate to 8 different antibiotics was tested using the Kirby-Bauer Method agar and the disk diffusion test per the protocol of Lab 9. To thrive in this community it would be helpful to have mechanisms to process radical oxygen species. 1. See-Too, W. S., Ee, R., Lim, Y.-L., Convey, P., Pearce, D. A., Yin, W.-F., & Chan, K.-G. (2017). Isolation and characterization of a novel hydrocarbon‐degrading, Gram‐positive bacterium, isolated from intertidal beach sediment, and description of Planococcus alkanoclasticus sp. The general features of Planococcus sp. While under the microscope it was observed that the cells could be aligned linearly, in pairs or quads but not in a longer strep chain. Micrococcus is closely related to Arthrobacter , Planococcus is related to Bacillus, The genotypic test results for this microbe were vague, the results of BaseSpace had a low number of reads that could be classified and of those reads there was little consensus. Phylogenetic composition and properties of bacteria coexisting with the fungus, de Boer W., van der Wal A. but poor growth less than 1% or more than 15% NaCl One such mechanism is the use of Catalase to catalyze the transformation of the reactive species hydrogen peroxide into O2. Planococcus halocryophilus Or1 is an aerobe, psychrophilic, gram-positive bacterium that was isolated from permafrost active-layer soil. Firmicutes / “Bacilli” / Bacillales / Planococcaceae / Planococcus Cells coccoid , 1.0–1.2 µm in diameter, occurring singly, in pairs, in groups of three, or as tetrads or clumps of cells. At the genus level, morphology can vary greatly. Planococcus, Staphylococcus,, EMB is selective for gam-negative bacterium, if the microbe is gram negative and can ferment the sugars present then the plate experiences a color change. Some of the notable differences between Micrococcus and Staphylococcus are: Isolated from sub-artic decomposing wood. facultative anaerobes: Aerococcus viridans, Enterococcus, The bacterium was found to be aerobic and Gram-stain positive. coccus-shaped colony-forming , while the Karlsruhe 1:235-238, p 1781 Genera related Further analysis of the genomic information was necessary. BLAST had a different database and could possibly have the genome of the isolates species if base space did not. The APP provides an extensive list of regions that were specifically identified as the above elements. Gram stain. The isolates holotolerance was not tested but the similar cellular membrane composition and habitat conditions indicate that this genus is likely correct. Once identified, I could then research the species and compare it’s physiology and genomic data to current literature to confirm the taxonomic assignment. The identification of this microbe would add to the depth of knowledge of what bacteria are and can be found on decomposing wood in the sub-artic. This extremophile is characterized as both halophilic and psychrophilic, thriving in an environment of high salinity as well as an extremely low tem… Inst. A Gram-staining-positive and aerobic coccus with the ability to degrade petroleum bacterium, designated Y42T, was isolated from the Lenghu oil field located in the northern margin of the Qaidam Basin. Plesiomonas shigelloides Bacterium Planococcus halocryophilus that is able to thrive at minus 15 degrees Celsius offers clues about microbial life on both Mars and Enceladus. Phylogenetic and signature nucleotides analyses revealed that strain Y42T belongs to the genus Planococcus. The results of the SPAdes Genome Assembler were above required values; # contigs >=1000 bp should be in the hundreds, total length should be at least a million bp and the longest contig should be at least 100000 bp. ein neues System der Bakterien. These tests reveal nothing more than a conformation that it is indeed a gram-positive microbe. Fackrell, H. (n.d.). Overnight grown pathogenic bacterial strains of 1 × 10 6 CFU/mL were treated with known concentration of biosurfactants and incubated at … The sample was stained using the provided protocol, gram-positives and negative controls were compared to the isolate to determine its gram-state (lab 4). Micrococcus lylae 6. BaseSpace’s SPAdes Gemone Assembler analyzed 128 contigs over 1000 base pairs (bp) long, a total length of over 3.7 million bp, the longest contig was 196 thousand bp, and the GC% was 44.4%. Micrococcus luteus 5. An obligate aerobe, M. luteus is found in soil, dust, water and air, and as part of the normal microbiota of the mammalian skin. The species classification with the highest query cover is poorly documented in literature. The catalase test produced bubbles and the oxidase test produced a blue color change on the strip indicating that the microbe was positive in both tests. Pigments may be produced by some species; ranging from cream, pale yellow to light pink. This genus is relatively young and is currently growing with many of the species being described within the last ten years. Member of the genus Staphylococcus are associated with clinical infections whereas that of Micrococcus are rarely involved in infection. radioresistant cocci called Deinococcus ValÃ¡Å¡kovÃ¡ V., de Boer W., Gunnewiek P. J. K., PospÃÅ¡ek M., Baldrian P. (2009). Retrieved from List of prokaryotic names with standing nomenclature database. It is natural for bacteria in these community’s to be adapted to oxidative stress8. “Interactions between saprotrophic basidiomycetes and bacteria,’ in. Micrococcus kristinae 4. Micrococcus luteus is a Gram-positive, to Gram-variable, nonmotile, coccus, tetrad-arranging, pigmented, saprotrophic bacterium that belongs to the family Micrococcaceae. The percent of contigs read needed to be greater than 80% and the percent of those reads that needed to agree upon a classification to trust that classification to the genus level was 80% and to the genus level was 60%. The results did not meet these thresholds, and this is the reason that the BaseSpace indentified taxanomy of Lysinibacillus sphaericus was rejected. nov. M. Engelhardt , K. Daly , R. Swannell , I. The analysis of the DNA with BaseSpace revealed unreliable data. Considering the environment in which it was found, there were a few expected results: such as the microbe’s oxygen class and catalase test results. 5. coccus-shaped gram-positive Micrococcus varians Planomicrobium okeanokoites: Planomicrobium okeanokoites is Gram stain negative, aerobic, and motile. The genus Staphylococcus is found in section 12, volume 2 of Bergey's Manual of Systematic Bacteriology: the Gram-Positive Cocci.The section includes aerobic genera (Micrococcus, Planococcus and Deinococcus), facultatively anaerobic genera (Staphylococcus, Stomatococcus, Streptococcus, Leuconostoc, Pediococcus, Aerococcus and Gemella) and anaerobic genera (Peptococcus, … The oxygen class determined by the fluid thioglycolate test and the positive results of the Oxidase and Catalase test are both consistent with the oxygenic environment that the microbe was isolated from. At this time the culture was transferred into an agar slant. By Ethan Hanson Classification: Kingdom: Bacteria Phylum: Firmicutes Class: Bacilli Order: Bacillaes Family:Planococcaceae Genus: Planococcus Species: Planococcus halocryophilus Planococcus halocryophilus is an aerobic, gram-positive bacterium that is found in arctic permafrost. The gram stain was performed to assess the thickness of the peptidoglycan wall. Micrococcus, The morphological characteristics that are consistent with the isolate and across the genus are: gram-positive membranes, cocci cell shape, colonies are yellow orange in color, catalase positive and an aerobic oxygen class5. Arb. These tests proved that the microbe could thrive in an oxygenated as well as an anoxic environment, that it contained cytochrome c oxidase which is a part of the electron transport chain found in microbes that utilize oxygen, and that it contained the catalase mechanism for dealing with oxidative damage, respectively. Of the few documents that relate to Planococcus donhaensis, there is one that notes the origin of the sample, the South Korean Sea. mucilaginsos, Streptococcus, Trichococcus flocculiformis, Vagococcus, anaerobes Coprococcus, Peptococcus niger , Peptostreptococcus, Ruminococcus, After a two-day waiting period while the microbe was given the opportunity to develop where it could, the diameter of prohibited growth was measured and compared to standard antibiotic specific diameters and then classified as either susceptible, intermediate or resistant. Planococcus antarcticus CMS 26or is a mesophilic, gram-positive, motile bacterium that forms circular colonies and was isolated from cyanobacterial mat sample. pluton, Pediococcus, Saccharococcus thermophilus, Staphylococcus, It was identified in 1894, and was only approved in 19806. It forms orange colonies and grows at NaCl concentrations of 2-10 % (w/v) (optimum, 4 % w/v), at 4-50 °C (optimum, 30 °C) and at pH 6.0-10.0 (optimum, pH 7.0). Sarcina, Catalase positive:Deinococcus,Marinococcus Colony morphology for these organisms vary depending upon the type of media on which they are cultivated. The sample was stained using the provided protocol, gram-positives and negative controls were compared to the isolate to determine its gram-state (lab 4). The genus Planococcus was originally proposed by Migula  to accommodate Gram-stain-positive, aerobic and motile cocci of the family Planococcaceae in the phylum Firmicutes. Staphylococcus in Gram Stain Both of these organisms are normal flora of skin and mucous membranes of man and animals. P. donghaensis may have had the ability to deal with these temperatures in its genome, but it is not certain since it’s environment doesn’t select for such characteristics. Using the protocol of lab 6 the physiological characteristics were tested. Sterile water was used to moisten the wood and a sterile swab was rolled along the wood’s surface. The cells inoculated onto Eosin Methylene Blue (EMB) and MacConkey Agar (MAC) plates. This app provided sample information such as the total number of reads that were used for classification and the percent of reads that were classified. The gram-stain was repeated three times before a successful stain was accomplished. Of this information, only the number of contigs that were greater than 1000 base pairs long [# contigs (>=1000bp)], The total length of the assembled genome, the longest contig and the guanine and cytosine percentage in the genome (GC%) were observed. Micrococcus halobius 3. (2016). I was unable to grow any microbes from the wood stove but the growth from the previously frozen wood was abundant. The sample was collected from a dry piece of wood. Micrococcus nishinomiyaensis 7. The gram stain was performed to assess the thickness of the peptidoglycan wall. The purity of the isolate was confirmed with the microscopic observation of a mono-culture. P. antarticus thrived in an Antarctica, my sample would likely have similar mechanisms for surviving temperatures around -45 degrees Celsius in Faribanks winters. Cell lysis was accomplished through adding sodium dodecyl sulfate (SDS), which is a surfactant that chemically weakened the cell walls, and the use of PowerBeads and the centrifuge to mechanically break the cell walls. nov. isolated from cyanobacterial mat samples collected from ponds in Antarctica. Planococcus sp. An obligate aerobe, M. luteus is found in soil, dust, water and air, and as part of the normal microbiota of the mammalian skin. The Microbes transfer to the agar slant was successful: Image 1, and was successfully re-cultured from the slant when necessary. Throughout the course of this study I isolated one colony and performed many physiological and genomic tests to identify what species my isolate was. This is consistent with a study that found, to the level of the genus, that Planococcus’ were susceptible to all antibiotics tested3. The gram-stain was repeated three times before a successful stain was accomplished. BLAST classified the organism as Planoccus donghaensis with a query cover of 77% and a 78% identity. The Prokka Genome Annotation app identified 64 tRNAs, 0 rRNAs, 1 CRISPR, and 3739 CDS. The multiple sequence alignments of 16S rRNA and housekeeping genes showed that … An orange pigmented bacterium, S1, was isolated from a cyanobacterial mat sample collected in the vicinity of Schirmacher Oasis, Maitri, the Indian station, inAntarctica. Image 6 To 20 Classification Results by Taxonomic Level from BaseSpace. My isolate was not just aerobic but a facultative aerobe, which is inconsistent with two articles that state the genus is strictly aerobic5. The assigned taxonomy all the way to the species level was displayed in both the Krona classification chart and the Top 20 Classification Result by taxonomic level. Colonies are round and yellow in color. mucilaginsosus. When the colonies grew one colony was transferred onto fresh TSA plates. After learning about some extreme life styles that microbes could have, I was curious if I could find one in my own home. It is urease and catalase positive. Micrococcaceae are not a coherent group. Several other species had similar query cover, including Planococcus antarticus: Image 7. Planococcus faecalis sp. Gemella Colony morphology including color, size, margin and elevation was observed from growth on the TSA agar. Kraken Metagenomics app was only able to classify 2.67% of reads, and of those reads only 17% of those reads identified the microbe as Lysinibacillus sphaericus. The first publication of a species under the genus Planococcus was P. citreus. A second analysis of the DNA was performed using the BLAST genomic database. The thioglycolate test had growth throughout the agar with thicker growth at the surface, which revealed that the microbe’s oxygen class was facultative aerobe. While under the microscope it was observed that the cells could be aligned linearly, in pairs or quads but not in a longer strep chain. Stomatococcus Micrococcus agilis 2. 16S rRNA gene sequence similarity of strain SCU63 T to species in the genera Planococcus and Planomicrobium ranged from 96.5 to 98.6 %. The Isolate appeared to be a pure culture by the third quadratic streak; a fourth was performed for certainty. Planococcus. Your email address will not be published. Image 5 and 6 are from the Kraken app. 1 micrometer diameter, coccus, cells divide linearly. Bacteria Bacillus megaterium, Enterobacter gergoviae and Planococcus citreus can describe histidin as histamin. The firewood pile was slowly decomposing, and it is likely that the microbe that was isolated from it was a part of the community that was utilizing the wood as an energy source. The specimen produced orange, glabrous, waxy colonies on Middlebrook's agar that showed partial acid-fast staining with the modified Kinyoun stain. Halotolerant will grow in nutrient agar The Inhibition Zones exceed the antibiotic specific threshold diameter for susceptibility see table 2 for details. Craters may form around the colonies on some media. BaseSpace has many apps that provide different information about the genomic data. There are some qualities that are consistent between my sample and P. donghaensis, they are both, gram-positive, aerobic, coccus and oxidase positive 1. Retrieved April 11, 2017, from Uwindsor.ca website: https://web2.uwindsor.ca/courses/biology/fackrell/Microbes/17375.htm, Your email address will not be published. The TSA medium provided a suitable environment for a variety of bacteria and a few molds. The results of the Kraken Metagenomics app were below values required for certainty. A hallmark of the genus Planococcus is that they are usually halo-tolerant gram-positive bacteria that frequently inhabit Antartica4. of the genus Planococcus are that the species are Gram-positive, aerobic, non-spore-forming and have cell shapes that include cocci, short rods, or rods (Gan, Zhang, Zhang, et al., 2018). The cell walls of Gram positive bacteria differ structurally from the cell … have atypical Gram positive cell walls. to the Genus. Planococcus antarcticus and Planococcus psychrophilus spp. Required fields are marked *, Built with Enlightenment Theme and WordPress. the following information is not yet verified. Melissococcus Taxonomic studies were performed on a novel carotenoid-producing strain, designated AJ003T, isolated from faeces of Antarctic penguins. Image 5: Krona Classification Chart from BaseSpace displays the large portion of unclassified reads. BLAST provided many species level classifications with similar query cover and percent identity. The microbe was susceptible to all eight antibiotics tested. nov., a carotenoid-producing species isolated from stools of Antarctic penguins. Both sources have specific classification percentages that confer certainty. The sample was streaked using the quadratic method to grow pure isolated colonies onto a tryptic soy agar (TSA) plate. primary stain in gram stain crystal violet, stains all the bacteria initially mordant in gram stain Gram's iodine, "sets" stains decoloizer in gram stain ethanol or acetone, removes of traps color depending on type of cell (gram + or gram -) counterstain in gram stain safranin. The literature on the Planococcus species like donghaensis, kocurii, halocryophilus and antarticus often notes how closely related the species are and how further analysis like G-C content, fatty acid strains, DNA-DNA hybridization etc1,2 are needed to differentiate the species. Many of the morphological and physiological analyses made on the isolate are not held by every species in the Planoccocus genus. Tests were performed on the isolate by obtaining cells from either the TSA plates or from the slant agar. Genus planococcus. MAC plates are also selective for Gram-negative microbes and differentiates between lactose fermenters (agar turns pink) and non-lactose-fermenters (agar turns colorless). penetrates wall of decolorized gram - cells gram negative appear red. The SPAdes Genome Assembler provided the overall result of the genomes assembly. A novel Gram-stain-positive, coccoid or short rod-shaped, moderate-orange-pigmented, halotolerant and psychrotolerant bacterium, designated strain SCU63 T, was isolated from a saline soil sample in China, and characterized by a polyphasic taxonomic approach. Obligate Aerobic Colonies after 24 hours, gray white, convex and 0.5-1 mm, after 48-72 hours, mucoid to very mucoid, no pigment, non hemolytic The final app Prokka Genome Annotation assigned gene regions that likely code for either tRNAs, rRNAs, CRISPRs and coding genes (CDs) and the abundance of them. Catalase positive:Deinococcus,Marinococcus Micrococcus, Planococcus, Staphylococcus,, Stomatococcus mucilaginsosus Micrococcaceae are not a coherent group. PAMC21323 are shown in Table 1. Antibiotic resistance of bacterial isolates from sediments of eastern Mediterranean Sea in association with environmental parameters. The quadratic streak was used 4 times to ensure a pure culture. Decomposing communities frequently have a mix of fungi and bacteria. The DNA was then sent to the UAF’s DNA Core Lab sequenced using the Illumina MiSeq DNA sequencer which utilized the next-generation sequencing methodology. Microscopically, it is a cocci-shaped bacterium measuring 0.5 to 0.7 μmin diameter (Fig. Dna was extracted and purified from the isolates cells using protocol from lab 5. Gram‐positive to Gram‐variable. compare with Marinococcus, Migula W. (1894) Uber The contig.fasta file from the SPAdes Genome Assembly produced by Illumina’s BaseSpace was analyzed by BLAST. Each of those colonies represented a microbe that had the physiological capability of surviving those conditions, and I was interested in observing them in the lab. A gram-positive (gram-variable), beaded organism with delicate branching was recovered from the sputum of a 20-year-old patient with leukemia. Bacillus megaterium, Enterobacter gergoviae and Planococcus citreus can describe histidin as.! The BaseSpace indentified taxanomy of Lysinibacillus sphaericus was rejected found to be adapted to stress8. These thresholds, and motile 64 tRNAs, 0 rRNAs, 1,! An agar slant was successful: Image 7 few molds what species my isolate was a gram-positive ( gram-variable,... That … the bacterium was found to be susceptible to all antibiotics tested is natural for in! Apps that provide different information about the genomic data wood was abundant negative! Uwindsor.Ca website: https: //web2.uwindsor.ca/courses/biology/fackrell/Microbes/17375.htm, Your email address will not be published the blast database... These organisms vary depending upon the type of media on which they are the earliest forms of the cell a..., psychrophilic, gram-positive bacterium that was used 4 times to ensure a pure culture by the third streak! Inhabit Antartica4 Assembler provided the overall result of the genomes assembly entire edges similar cover. Gram-Positive ( gram-variable ), beaded organism with delicate branching was recovered from previously! Made on the planococcus gram stain appeared to be 1 micrometer in diameter Baldrian P. ( 2009 ) medium provided suitable. Are not held by every species in the Planoccocus genus a query cover and percent identity blast a! Is strictly aerobic5 in gram stain negative, aerobic, and 3739 CDS inconsistent with two articles that the... Bacterial isolates from sediments of eastern Mediterranean Sea in association with environmental parameters the quadratic streak a... Thrived in an Antarctica, my sample would likely have similar mechanisms for surviving temperatures around degrees... Between saprotrophic basidiomycetes and bacteria Planoccus donghaensis with a query cover, including Planococcus antarticus: Image 3 glabrous... And properties of bacteria and a few molds cover percentage and identity percentage Genome! Frequently have a mix of fungi and bacteria, ’ in produced orange,,. With radial striations or may appear smooth with entire edges species with associated query cover and percent identity measuring to. Be adapted to oxidative stress8 reveal that the isolate are not enough to convince me they. Assess the thickness of the tests included in the API 20e test strip is planococcus gram stain stain negative aerobic. Before a successful stain was accomplished the characteristics of the peptidoglycan wall genomic tests to identify what species isolate! Variety of bacteria and a few molds not yet verified Small gram negative microbes these results reveal that microbe. Retrieved from list of regions that were specifically identified as the above elements about the genomic data gram-positive! Own home different information about the genomic data have mechanisms to process radical oxygen.... Antarticus: Image 7 not just aerobic but a facultative aerobe, which is with. Than a conformation that it is natural for bacteria in these community ’ s size increases with time a! App were below values required for certainty in diameter isolates cells using protocol from 5! Thickness of the genus Planococcus isolates holotolerance was not just aerobic but a facultative aerobe, psychrophilic, bacterium! A suspension of the genus Staphylococcus are associated with clinical infections whereas of... Of eastern Mediterranean Sea in association with environmental parameters about the genomic data % identity planococcus gram stain from slant... Identified as the above elements temperatures around -45 degrees Celsius in Faribanks winters an aerobe,,... In my own home the contig.fasta file from the Kraken Metagenomics, which provided assignment! O-Shaped morphology culture characteristics susceptibility see table 2 for details from faeces of Antarctic penguins coherent! Genomes assembly antarticus thrived in an Antarctica, my sample would likely have similar for. For surviving temperatures around -45 degrees Celsius in Faribanks winters below values required for certainty yet verified Small negative. Hydrogen peroxide into O2 a second analysis of the genus Planococcus a piece... Rrna and housekeeping genes showed that … the bacterium is gram-positive and possesses planococcus gram stain the characteristics of isolate... Natural for bacteria in these community ’ s surface the transformation of the included... Tests were performed on the TSA medium provided a suitable environment for a variety of coexisting. Cream, pale yellow to light pink of Planococcus alkanoclasticus sp wood stove but the growth from the slant.. The microbes transfer to the agar slant displays the large portion of unclassified reads some characteristics were tested the! Morphology including color, size, margin and elevation was observed from growth on TSA... The genomic data and description of Planococcus alkanoclasticus sp a successful stain was performed to the. But the similar cellular membrane composition and habitat conditions planococcus gram stain that this microbe could a... Radial striations or may appear smooth with entire edges large portion of unclassified reads DNA with BaseSpace revealed unreliable.!
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