Microbial biodiversity of Tang and Pirgal mud volcanoes and evaluation of bio-emulsifier and bio-demulsifier activities of Capnophile bacteria

The data presented in this article is related to the Master thesis; entitled “Survey Aerobic Microbial Diversity Mud Volcanoes in Chabahar and Khash Ports in Southern Iran” by the first author of this article, year 2011, Islamic Azad University, Iran (reference number (Parsia, 2011) [1] of this article). This article shows microbial biodiversity and evaluates bio-emulsifier and bio-demulsifier abilities of capnophile isolates, in order to introduce a superior isolate for the Microbial Enhanced Oil Recovery (MEOR) process in the petrochemical industry.


a b s t r a c t
The data presented in this article is related to the Master thesis; entitled "Survey Aerobic Microbial Diversity Mud Volcanoes in Chabahar and Khash Ports in Southern Iran" by the first author of this article, year 2011, Islamic Azad University, Iran (reference number (Parsia, 2011) [1] of this article). This article shows microbial biodiversity and evaluates bio-emulsifier and biodemulsifier abilities of capnophile isolates, in order to introduce a superior isolate for the Microbial Enhanced Oil Recovery (MEOR) process in the petrochemical industry.
& 2017 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). This data would be valuable for further studies to optimize the bio-emulsifier and bio-demulsifier activities of recognized isolates.

Specifications
Used direct molecular identification methods to recognize species and compare with currently culture and biochemical methods.

Data
The dataset used in this article provides information on the microbial biodiversity of both mud volcanoes as well as the bio-emulsifier and bio-demulsifier activities of capnophile isolates, in order to use them in the Microbial Enhanced Oil Recovery (MEOR) process of the petrochemical industry. Presentation of data in this article is described in Table 1. Table 1 Presentation of data.

Presented data Tables
Name of group and number of microbial isolates from Tang and Pirgal mud volcanoes Table 2 Biochemical identification of gram-negative bacteria Table 3 Biochemical identification of spore forming gram-positive rods Table 4 Biochemical identification of irregular colony, non-sporing, gram-positive rod strains with different catalase tests ( þ or -)

Tables 5 and 6
Biochemical identification of regular colony, non-sporing, gram-positive rod strains with different catalase tests ( þ or -)

Tables 7 and 8
Biochemical identification of non-sporing gram-positive coccus strains with different catalase tests ( þ or -)

Tables 9 and 10
Identification of superior bio-demulsifier capnophile isolates based on degree of demulsification, followed by surface tension measurement and biochemical and molecular identification Tables 11, 12, and 13 Identification of superior bio-emulsifier capnophile isolates based on degree of emulsification, followed by surface tension measurement and biochemical and molecular identification Tables 14, 15 and 16    Table 4 Biochemical tests for the identification of spore forming gram positive rods.

Table 8
Biochemical tests for the identification of regular colony, non-sporing, gram positive rod strains, catalase negative.

Isolate Test
Oxygen Motility Growth at 35°C LV reaction Citrate utilization B: Mesophilic facultative anaerobic; C: Capnophile; A/A: Acid/acid. All isolates showed 98% o similarity to Erysipelothrix.sp.

Table 9
Biochemical tests for the identification of non-sporing, gram positive coccus strains, catalase positive.

Isolate Test
Oxygen Motility Acid fast staining CAMP OF LV reaction VP reaction Citrate utilization Oxidase

Table 10
Biochemical tests for the identification of non-sporing, gram positive coccus strains, catalase negative.

Isolate Test
Oxygen Motility Acid fast staining LV reaction Citrate utilization Oxidase Vancomycin sensitive Growth at 10°C

Experimental design, materials and methods
In the summer of 2011, sampling was performed at Tang and Pirgal mud volcano craters, in aseptic conditions, using sterile plastic pipes (in sizes of 5, 10, 15 and 30 cm) [1].
Each sample was diluted in 9cc strilled Ringer's solution. Next, 1 cc of the solution was added to 9 cc of strilled nutrient broth medium and incubated at 30°C for 48 h. Each microbial group used specific conditions, such as medium culture (MC), temperature (tem) and time (T) of incubation [1]. For biochemical identification, isolates were classified based on their colony shape, morphology and gram-stain. They were then identified using tests for gram negative bacteria, gram positive nonsporing and spore-forming bacilli (A colour Atlas of Bacillus species) and cocci bacteria based on table and diagram references [1][2][3][4].
The bio-emulsifier test used the Francy method (year 1991) and assessed their stabilizing emulsification capacity (degree 0-4) [5,6]. In the bio-demulsifier test, 1 ml from Erlenmeyer flasks was added to tubes containing stable emulsions of water/diesel and diesel/water. They were then properly vortexed and incubated at 30°C for the assessment of demulsification degree (0 to 5). The surface tensions of superior isolates were measured by Tensiometer (TD1C LAUDA) [7,8]. Superior isolates were identified with molecular tests. Their genomes were extracted by kit. The universal primers used to amplify 16S rDNA, were 27 F(5′ AGA GTT TGA TCC TGG CTC AG 3′) and 1492 R(5′ CGG TTA CCT TGT TAC GAC TT 3′). These amplified a 1500-base pair region of the 16S rDNA gene. The amplified DNA was visualized by gel electrophoresis and sequenced. A 16S rDNA sequence was analysed using Chromas LITE. The most similar bacterial species was found in the GenBank using BLAST search. Neighbours joining phylogenetic trees were constructed based on 16S rDNA sequences using Clus-talW [1].