scholarly journals Survival of the Halophilic Archaeon Halovarius luteus after Desiccation, Simulated Martian UV Radiation and Vacuum in Comparison to Bacillus atrophaeus

2020 ◽  
Vol 50 (3-4) ◽  
pp. 157-173 ◽  
Author(s):  
Niloofar Feshangsaz ◽  
Farid Semsarha ◽  
Saeed Hesami Tackallou ◽  
Kamran Nazmi ◽  
Euan P. Monaghan ◽  
...  
Keyword(s):  
Uv Radiation ◽  
High Performance ◽  
Salt Lake ◽  
Protein Profile ◽  
Space Environment ◽  
Most Probable Number ◽  
Bacillus Atrophaeus ◽  
Probable Number ◽  
Phosphate Buffered Saline ◽  
Spore Forming Bacteria

AbstractExtraterrestrial environments influence the biochemistry of organisms through a variety of factors, including high levels of radiation and vacuum, temperature extremes and a lack of water and nutrients. A wide variety of terrestrial microorganisms, including those counted amongst the most ancient inhabitants of Earth, can cope with high levels of salinity, extreme temperatures, desiccation and high levels of radiation. Key among these are the haloarchaea, considered particularly relevant for astrobiological studies due to their ability to thrive in hypersaline environments. In this study, a novel haloarchaea isolated from Urmia Salt Lake, Iran, Halovarius luteus strain DA50T, was exposed to varying levels of simulated extraterrestrial conditions and compared to that of the bacteria Bacillus atrophaeus. Bacillus atrophaeus was selected for comparison due to its well-described resistance to extreme conditions and its ability to produce strong spore structures. Thin films were produced to investigate viability without the protective influence of cell multi-layers. Late exponential phase cultures of Hvr. luteus and B. atrophaeus were placed in brine and phosphate buffered saline media, respectively. The solutions were allowed to evaporate and cells were encapsulated and exposed to radiation, desiccation and vacuum conditions, and their post-exposure viability was studied by the Most Probable Number method. The protein profile using High Performance Liquid Chromatography and Matrix Assisted Laser Desorption/Ionization bench top reflector time-of-flight are explored after vacuum and UV-radiation exposure. Results showed that the change in viability of the spore-forming bacteria B. atrophaeus was only minor whereas Hvr. luteus demonstrated a range of viability under different conditions. At the peak radiation flux of 105 J/m2 under nitrogen flow and after two weeks of desiccation, Hvr. luteus demonstrated the greatest decrease in viability. This study further expands our understanding of the boundary conditions of astrobiologically relevant organisms in the harsh space environment.

scholarly journals The affect of the space environment on the survival ofHalorubrum chaoviatorandSynechococcus(Nägeli): data from the Space Experiment OSMO on EXPOSE-R

2014 ◽  
Vol 14 (1) ◽  
pp. 123-128 ◽  
Author(s):  
R. L. Mancinelli
Keyword(s):  
Survival Rate ◽  
Uv Radiation ◽  
Molecular Probes ◽  
Low Earth Orbit ◽  
Space Environment ◽  
Dust Particles ◽  
Most Probable Number ◽  
Earth Orbit ◽  
Probable Number ◽  
Space Vacuum

AbstractWe have shown using ESA's Biopan facility flown in Earth orbit that when exposed to the space environment for 2 weeks the survival rate ofSynechococcus(Nägeli), a halophilic cyanobacterium isolated from the evaporitic gypsum–halite crusts that form along the marine intertidal, andHalorubrum chaoviatora member of the Halobacteriaceae isolated from an evaporitic NaCl crystal obtained from a salt evaporation pond, were higher than all other test organisms exceptBacillusspores. These results led to the EXPOSE-R mission to extend and refine these experiments as part of the experimental package for the external platform space exposure facility on the ISS. The experiment was flown in February 2009 and the organisms were exposed to low-Earth orbit for nearly 2 years. Samples were either exposed to solar ultraviolet (UV)-radiation (λ > 110 nm or λ > 200 nm, cosmic radiation (dosage range 225–320 mGy), or kept in darkness shielded from solar UV-radiation. Half of each of the UV-radiation exposed samples and dark samples were exposed to space vacuum and half kept at 105pascals in argon. Duplicate samples were kept in the laboratory to serve as unexposed controls. Ground simulation control experiments were also performed. After retrieval, organism viability was tested using Molecular Probes Live–Dead Bac-Lite stain and by their reproduction capability. Samples kept in the dark, but exposed to space vacuum had a 90 ± 5% survival rate compared to the ground controls. Samples exposed to full UV-radiation for over a year were bleached and although results from Molecular Probes Live–Dead stain suggested ~10% survival, the data indicate that no survival was detected using cell growth and division using the most probable number method. Those samples exposed to attenuated UV-radiation exhibited limited survival. Results from of this study are relevant to understanding adaptation and evolution of life, the future of life beyond earth, the potential for interplanetary transfer of viable microbes via meteorites and dust particles as well as spacecraft, and the physiology of halophiles.

Evaluation of the Compact Dry VP Method for Screening Raw Seafood for Total Vibrio parahaemolyticus

2009 ◽  
Vol 72 (1) ◽  
pp. 169-173 ◽  
Author(s):  
HIDEMASA KODAKA ◽  
HAJIME TERAMURA ◽  
SHINGO MIZUOCHI ◽  
MIKAKO SAITO ◽  
HIDEAKI MATSUOKA
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Cold Water ◽  
Correlation Coefficients ◽  
Water Soluble ◽  
Coliform Bacteria ◽  
Most Probable Number ◽  
Probable Number ◽  
Different Types ◽  
Phosphate Buffered Saline ◽  
Vibrio Spp

Compact Dry VP (CDVP) is a ready-to-use method for enumerating Vibrio parahaemolyticus in food. The presterilized plates contain a culture medium comprising peptone, NaCl, bile salts, antibiotics, chromogenic substrates, and polysaccharide gum as a cold water–soluble gelling. After diluting raw seafood samples in a phosphate-buffered saline solution, a 1-ml aliquot was inoculated onto the center of the plate and allowed to diffuse by capillary action. Blue-green colonies forming on the plates were counted after 18 to 20 h of incubation at 35°C. A total of 85 V. parahaemolyticus strains (62 tdh+ strains and 23 tdh− strains) were studied for inclusivity, 81 (95.3 %) of which produced blue-green colonies. When 97 strains (14 strains of Vibrio spp., 33 strains of coliform bacteria, and 50 strains of noncoliform bacteria) were assessed for exclusivity, 10 strains of Vibrio spp. produced non–blue-green colonies, and 87 strains failed to grow. The CDVP and U.S. Food and Drug Administration Bacteriological Analytical Manual (FDA-BAM) methods were compared with the use of four different types of raw seafood that were inoculated with four different V. parahaemolyticus strains. For raw tuna and oysters, the FDA-BAM colony lift method was used, whereas the FDA-BAM most-probable-number method was used for salmon and scallop. The linear correlation coefficients between the CDVP and FDA-BAM methods were 0.99 for fresh raw tuna, 0.95 for fresh raw oysters, 0.95 for frozen raw salmon, and 0.95 for frozen raw scallops. These results suggest that the CDVP method is useful for screening raw seafood for V. parahaemolyticus.

scholarly journals Farm Silage Facilities and Their Management for the Prevention of Anaerobic Bacteria Spore Contamination in Raw Milk

Dairy ◽  
2021 ◽  
Vol 2 (3) ◽  
pp. 500-514
Author(s):  
Fabio Abeni ◽  
Rosanna Marino ◽  
Francesca Petrera ◽  
Giulia Segati ◽  
Andrea Galli ◽  
...  
Keyword(s):  
Anaerobic Bacteria ◽  
Raw Milk ◽  
Most Probable Number ◽  
Facilities Management ◽  
Probable Number ◽  
Factors Affecting ◽  
Po Valley ◽  
Face Width ◽  
Bulk Milk ◽  
Spore Forming Bacteria

At feed-out, aerobic spoilage of silage enables an increase in anaerobic spore-forming bacteria (ANSB) that may enter the total mixed ration (TMR). The aim of our study was to understand whether in hot summers the silage structures and management may affect the level of ANSB in milk for long-ripening cheese production. A survey of silage facilities, management, and their relationships with silage, TMR, feces, and milk ANSB most probable number (MPN) content was conducted in the Po Valley during summer months. Silo type did not affect the mean ANSB, but only the wideness of their value distributions, with a narrow range for bags and a wider range for bunkers. The unloading equipment affected the ANSB count; the front-end loader with cutter was associated with a lower ANSB count—probably as a result of the reduced surface left after daily silage removal. Silo length and daily removed face width were the main factors affecting contamination of silage by spore-forming bacteria during summer, with longer silos and wider surface removal reducing ANSB contamination—probably as a consequence of reduced aerobic spoilage at the silage surface. The silage contamination by spore-forming bacteria within a log10 2 MPN g−1 allowed a low concentration of spore-forming bacteria at the farm bulk milk tank level. Fecal ANSB levels did not factor into the regression that explains the ANSB in farm milk. It has been found that silage facilities’ features and their management are an important first step to reduce the extent of ANSB contamination at the farm level.

Glucose and plant exudate enhanced enumeration of bacteria capable of degrading polycyclic aromatic hydrocarbons

2011 ◽  
Vol 57 (12) ◽  
pp. 1067-1072 ◽  
Author(s):  
John C. Thomas ◽  
Robert T. Dabkowski
Keyword(s):  
Root Exudates ◽  
Contaminated Soils ◽  
High Performance ◽  
Most Probable Number ◽  
Plant Interactions ◽  
Probable Number ◽  
Chromatography Analysis ◽  
Pah Degradation ◽  
Polycyclic Aromatic ◽  
Low Levels

Enumerating environmental microbial isolates capable of polycyclic aromatic hydrocarbon (PAH) degradation can provide insight into the microbe–plant interactions that facilitate PAH removal. We examined a known PAH degrader ( Pseudomonas putida G7), a nondegrader ( Agrobacterium tumefaciens LBA4404), and several microorganisms isolated from the environment by using a PAH cocktail in an enumeration medium with or without 0.025% (m/v) glucose and (or) root exudates. Compared with the standard most probable number (MPN), the addition of glucose and root exudates in a modified MPN method resulted in a 3- to 11-fold enhancement of PAH degraders being enumerated among microorganisms found in PAH-contaminated soils. High-performance liquid chromatography analysis verified that PAH levels were reduced using this modified enumeration method. Low levels of glucose, perhaps in concert with other materials in exudates, may promote microbial metabolism, thereby enhancing PAH degradation.

Estimation of Methanogen Biomass by Quantitation of Coenzyme M

1999 ◽  
Vol 65 (12) ◽  
pp. 5541-5545 ◽  
Author(s):  
Dwayne A. Elias ◽  
Lee R. Krumholz ◽  
Ralph S. Tanner ◽  
Joseph M. Suflita
Keyword(s):  
Liquid Chromatography ◽  
Environmental Samples ◽  
High Performance ◽  
Methanogenic Bacteria ◽  
Most Probable Number ◽  
Environmental Matrices ◽  
Coenzyme M ◽  
Simple Method ◽  
Probable Number ◽  
Pure Cultures

ABSTRACT Determination of the role of methanogenic bacteria in an anaerobic ecosystem often requires quantitation of the organisms. Because of the extreme oxygen sensitivity of these organisms and the inherent limitations of cultural techniques, an accurate biomass value is very difficult to obtain. We standardized a simple method for estimating methanogen biomass in a variety of environmental matrices. In this procedure we used the thiol biomarker coenzyme M (CoM) (2-mercaptoethanesulfonic acid), which is known to be present in all methanogenic bacteria. A high-performance liquid chromatography-based method for detecting thiols in pore water (A. Vairavamurthy and M. Mopper, Anal. Chim. Acta 78:363–370, 1990) was modified in order to quantify CoM in pure cultures, sediments, and sewage water samples. The identity of the CoM derivative was verified by using liquid chromatography-mass spectroscopy. The assay was linear for CoM amounts ranging from 2 to 2,000 pmol, and the detection limit was 2 pmol of CoM/ml of sample. CoM was not adsorbed to sediments. The methanogens tested contained an average of 19.5 nmol of CoM/mg of protein and 0.39 ± 0.07 fmol of CoM/cell. Environmental samples contained an average of 0.41 ± 0.17 fmol/cell based on most-probable-number estimates. CoM was extracted by using 1% tri-(N)-butylphosphine in isopropanol. More than 90% of the CoM was recovered from pure cultures and environmental samples. We observed no interference from sediments in the CoM recovery process, and the method could be completed aerobically within 3 h. Freezing sediment samples resulted in 46 to 83% decreases in the amounts of detectable CoM, whereas freezing had no effect on the amounts of CoM determined in pure cultures. The method described here provides a quick and relatively simple way to estimate methanogenic biomass.

scholarly journals The Reaction of Cellulolytic and Potentially Cellulolytic Spore-Forming Bacteria to Various Types of Crop Management and Farmyard Manure Fertilization in Bulk Soil

Agronomy ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 772
Author(s):  
Jakub Dobrzyński ◽  
Paweł Stanisław Wierzchowski ◽  
Wojciech Stępień ◽  
Ewa Beata Górska
Keyword(s):  
Farmyard Manure ◽  
Crop Management ◽  
Total Carbon ◽  
Bulk Soil ◽  
Most Probable Number ◽  
Cellulolytic Bacteria ◽  
Probable Number ◽  
Microbiological Methods ◽  
The Impact ◽  
Spore Forming Bacteria

The ecology of cellulolytic bacteria in bulk soil is still relatively unknown. There is still only a handful of papers on the abundance and diversity of this group of bacteria. Our study aimed to determine the impact of various crop management systems and farmyard manure (FYM) fertilization on the abundance of cellulolytic and potentially cellulolytic spore-forming bacteria (SCB). The study site was a nearly 100-year-old fertilization experiment, one of the oldest still active field trials in Europe. The highest contents of total carbon (TC) and total nitrogen (TN) were recorded in both five-year rotations. The abundances of SCB and potential SCB were evaluated using classical microbiological methods, the most probable number (MPN), and 16S rRNA Illumina MiSeq sequencing. The highest MPN of SCB was recorded in soil with arbitrary rotation without legumes (ARP) fertilized with FYM (382 colony-forming units (CFU) mL−1). As a result of the bioinformatic analysis, the highest values of the Shannon–Wiener index and the largest number of operational taxonomic units (OTUs) were found in ARP-FYM, while the lowest in ARP treatment without FYM fertilization. In all treatments, those dominant at the order level were: Brevibacillales (13.1–43.4%), Paenibacillales (5.3–36.9%), Bacillales (4.0–0.9%). Brevibacillaceae (13.1–43.4%), Paenibacillaceae (8.2–36.9%), and Clostridiaceae (5.4–11.9%) dominated at the family level in all tested samples. Aneurinibacillaceae and Hungateiclostridiaceae families increased their overall share in FYM fertilization treatments. The results of our research show that the impact of crop management types on SCB was negligible while the actual factor shaping SCB community was the use of FYM fertilization.

scholarly journals Disinfection of an advanced primary effluent using peracetic acid or ultraviolet radiation for its reuse in public services

2014 ◽  
Vol 13 (1) ◽  
pp. 118-124 ◽  
Author(s):  
Flores R. Julio ◽  
Terres-Peña Hilario ◽  
Vaca M. Mabel ◽  
López C. Raymundo ◽  
Lizardi-Ramos Arturo ◽  
...  
Keyword(s):  
Uv Radiation ◽  
Peracetic Acid ◽  
Wastewater Reuse ◽  
Primary Treatment ◽  
Fecal Coliforms ◽  
Treated Wastewater ◽  
Most Probable Number ◽  
Probable Number ◽  
Bacteriological Quality ◽  
Treated Wastewater Reuse

The disinfection of a continuous flow of an effluent from an advanced primary treatment (coagulation-flocculation-sedimentation) with or without posterior filtration, using either peracetic acid (PAA) or ultraviolet (UV) radiation was studied. We aimed to obtain bacteriological quality to comply with the microbiological standard established in the Mexican regulations for treated wastewater reuse (NOM-003-SEMARNAT-1997), i.e., less than 240 MPN (most probable number) FC/100 mL. The concentrations of PAA were 10, 15, and 20 mg/L, with contact times of 10, and 15 min. Fecal coliforms (FC) inactivation ranged from 0.93 up to 6.4 log units, and in all cases it reached the limits set by the mentioned regulation. Water quality influenced the PAA disinfection effectiveness. An efficiency of 91% was achieved for the unfiltered effluent, as compared to 99% when wastewater was filtered. UV radiation was applied to wastewater flows of 21, 30 and 39 L/min, with dosages from 1 to 6 mJ/cm2. This treatment did not achieve the bacteriological quality required for treated wastewater reuse, since the best inactivation of FC was 1.62 log units, for a flow of 21 L/min of filtered wastewater and a UV dosage of 5.6 mJ/cm2.

Analysis of Coliform and Colifecal Total Pollution Test on Various Types of Drinking Water Using the MPN (Most Probable Number) Method

2020 ◽  
Vol 2 (2) ◽  
Author(s):  
Wanda Aulya ◽  
Fadhliani Fadhliani ◽  
Vivi Mardina
Keyword(s):  
Drinking Water ◽  
Fecal Coliform ◽  
Pathogenic Bacteria ◽  
Microbiological Quality ◽  
Coliform Bacteria ◽  
Most Probable Number ◽  
Inspection Method ◽  
Sample Number ◽  
Probable Number ◽  
Fecal Coliform Bacteria

Water is the main source for life and also the most severe substance caused by pollution. The mandatory parameters for determining microbiological quality of drinking water are total non-fecal Coliform bacteria and Coliform fecal (Escherichia coli). Coliform bacteria are a group of microorganisms commonly used as indicators, where these bacteria can be a signal to determine whether a water source has been contaminated by bacteria or not, while fecal Coliform bacteria are indicator bacteria polluting pathogenic bacteria originating from human feces and warm-blooded animals (mammals) . The water inspection method in this study uses the MPN (Most Probable Number) method which consists of 3 tests, namely, the presumption test, the affirmation test, and the reinforcement test. The results showed that of 15 drinking water samples 8 samples were tested positive for Coliform bacteria with the highest total bacterial value of sample number 1, 15 (210/100 ml), while 7 other samples were negative. From 8 positive Coliform samples only 1 sample was stated to be negative fecal Coliform bacteria and 7 other samples were positive for Coliform fecal bacteria with the highest total bacterial value of sample number 1 (210/100 ml).

Cemaran Eshericia coli dalam makanan laut yang beredar di pasar tradisional Kota Pontian

2015 ◽  
Vol 1 (1) ◽  
pp. 44
Author(s):  
Rafika Sari ◽  
Pratiwi Apridamayanti
Keyword(s):  
Most Probable Number ◽  
Probable Number

Latar Belakang: Makanan laut merupakan salah satu jenis makanan yang banyak dikonsumsi oleh masyarakat selain sebagai komoditi ekspor. Mengkonsumsi makanan laut yang telah terkontaminasi bakteri hidup atau toksin yang dihasilkannya dapat menyebabkan keracunan makanan. Tujuan penelitian ini adalah untuk mengetahui adanya kontaminasi bakteri koliform E.coli sebagai indikator pencemaran pada makanan laut dan memberikan informasi kelayakan dan keamanan konsumsi dari makanan laut di dua pasar tradisional terbesar di daerah Pontianak. Metode: Sampel yang digunakan adalah ikan, sotong dan udang. Penelitian terhadap sampel dilakukan menggunakan uji Most Probable Number (MPN) yang dilengkapi dengan uji biokimia untuk mengidentifikasi jenis bakteri pada sampel melalui penanaman bakteri pada media agar Lactose Broth (LB) dan Briliant Green Lactose Bile Broth (BGLB). Hasil: Hasil penelitian menunjukkan bakteri koliform E.coli terdeteksi pada 100% sampel dengan nilai MPN yang tidak memenuhi kriteria kelayakan konsumsi, yakni >3/g. Kesimpulan: Makanan yang ada tidak memenuhi kriteria kelayakan konsumsi.

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