Isolation, functional evaluation of probiotic properties and molecular identification of strains isolated from Iranian poultry's gut.

Background and Objectives: The use of probiotics as an alternative to antibiotics in poultry diets has attracted a lot of interest recently. In this context, the determination of probiotic characteristics was evaluated on several isolates obtained from Iranian poultry's gut. Materials and Methods: Probiotic characteristics such as hemolysis activity, acid, bile, gastric juice tolerate, in vitro adhesion assay, cell surface properties (hydrophobicity, auto-aggregation and co-aggregation) and antibiotic susceptibility test were evaluated. Finally, selected isolates identified molecularly after temperature-salt tolerance and extracellular enzyme activity (amylase, protease and cellulose). Results: Out of 362 strains isolated from native poultry in three geographical regions of Iran, nine strains (belong to Bacillus sp., Enterococcus sp., Pediococcus sp., Lactobacillus sp., Kluyveromyces sp.) showed resistance against gastrointestinal physiological conditions, desirable surface properties, ability to adhere to epithelial intestine cell line and antibiotic susceptibility. Also, these strains were discovered to be temperature-salt tolerant but, only a small number of them were able to produce hydrolase enzymes. Conclusion: According to the results, the selected strains can be introduced as native probiotic candidates for utilization in novel poultry feeds.

Bacillales: From Taxonomy to Biotechnological and Industrial Perspectives.

For a long time, the genus Bacillus has been known and considered among the most applicable genera in several fields. Recent taxonomical developments resulted in the identification of more species in Bacillus-related genera, particularly in the order Bacillales (earlier heterotypic synonym: Caryophanales), with potential application for biotechnological and industrial purposes such as biofuels, bioactive agents, biopolymers, and enzymes. Therefore, a thorough understanding of the taxonomy, growth requirements and physiology, genomics, and metabolic pathways in the highly diverse bacterial order, Bacillales, will facilitate a more robust designing and sustainable production of strain lines relevant to a circular economy. This paper is focused principally on less-known genera and their potential in the order Bacillales for promising applications in the industry and addresses the taxonomical complexities of this order. Moreover, it emphasizes the biotechnological usage of some engineered strains of the order Bacillales. The elucidation of novel taxa, their metabolic pathways, and growth conditions would make it possible to drive industrial processes toward an upgraded functionality based on the microbial nature.

Potential utilization of dairy industries by-products and wastes through microbial processes: A critical review.

The dairy industry generates excessive amounts of waste and by-products while it gives a wide range of dairy products. Alternative biotechnological uses of these wastes need to be determined to aerobic and anaerobic treatment systems due to their high chemical oxygen demand (COD) levels and rich nutrient (lactose, protein and fat) contents. This work presents a critical review on the fermentation-engineering aspects based on defining the effective use of dairy effluents in the production of various microbial products such as biofuel, enzyme, organic acid, polymer, biomass production, etc. In addition to microbial processes, techno-economic analyses to the integration of some microbial products into the biorefinery and feasibility of the related processes have been presented. Overall, the inclusion of dairy wastes into the designed microbial processes seems also promising for commercial approaches. Especially the digestion of dairy wastes with cow manure and/or different substrates will provide a positive net present value (NPV) and a payback period (PBP) less than 10 years to the plant in terms of biogas production.

Halomonas lysinitropha sp. nov., a novel halophilic bacterium isolated from a hypersaline wetland.

We carried out a polyphasic taxonomic study on a new halophilic strain designated 3(2)T, isolated from Meighan wetland, Iran. Cells of the novel strain were Gram-stain-negative, non-hemolytic, catalase- and oxidase-positive, rod-shaped, non-endospore-forming and motile. Cell growth occurred at 3-15 % NaCl (w/v; optimum, 5 %), pH 7.0-9.0 (optimum, pH 7.5-8.0) and 15-35 °C (optimum, 30 °C). 16S rRNA gene sequence comparisons confirmed the affiliation of strain 3(2)T to the class Gammaproteobacteria and the genus Halomonas with highest similarity to Halomonas daqiaonensis YCSA28T (98.4 %) and Halomonas ventosae Al12T (97.9 %). Experimental and in silico DNA-DNA hybridization values were 42.7 and 35.1% with H. daqiaonensis IBRC-M 10931T and 48 and 35.2% with H. ventosae IBRC-M 10566T, respectively, and indicated that they are different members of the same genus. The genome of the type strain was characterized by a size of 3.83 Mbp with 63 scaffolds and a G+C content of 64.8 mol%. Moreover, the average nucleotide identity values against H. ventosae Al12T and H. daqiaonensis YCSA28T were 88.8 and 88.5 %, respectively. The predominant respiratory quinone was Q-9 (92 %) with Q-8 (8 %) as a minor component. Major fatty acids were C16 : 0 cyclo, C19 : 0 ω8c, C16 : 1 ω7c and/or iso-C15:0 2-OH, C12 : 0 3-OH and C18 : 1 ω7c. The polar lipid profile of the strain contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphoaminoglycolipid and four unidentified phospholipids. According to our results, strain 3(2)T could be classified as a novel species in the genus Halomonas for which the name Halomonas lysinitropha sp. nov. is proposed. The type strain is 3(2)T (=IBRC M 10929T=LMG 29450T=CIP 111708T).

Planomicrobium iranicum sp. nov., a novel slightly halophilic bacterium isolated from a hypersaline wetland.

A novel strain, designated as MX6T was isolated from Meighan wetland, in the centre of Iran. The cells were Gram-stain-positive, motile, coccoid to rod-shaped, oxidase- and catalase-positive. The strain grew optimally at 35 °C, 3 % (w/v) NaCl and pH 7-7.5. A polyphasic taxonomic study was undertaken in order to characterize the strain in detail. The results of phylogenetic analysis based on 16S rRNA gene sequences indicated that MX6T represented a member of the phylum Firmicutes, family Planococcaceae, genus Planomicrobium, and showed the highest similarity with Planomicrobium flavidum ISL-41T (98.2 %) and Planomicrobium psychrophilum CMC 53orT (98.0 %). The main polar lipids of MX6T consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and seven unidentified phospholipids and its DNA G+C content was 45.5 mol%. Major cellular fatty acids were anteiso-C15 : 0, C16 : 1ω7c alcohol, iso-C14 : 0, iso-C15 : 0 and iso-C16 : 0 and the predominant respiratory quinone was Q-8 (62 %). Experimental DNA-DNA hybridization between MX6T and Planomicrobium flavidum IBRC-M 11047T was 20 %, supporting the differential taxonomic status of MX6T as representing a different taxon. All these data indicate that MX6T represents a novel species of the genus Planomicrobium, for which the name Planomicrobium iranicum sp. nov. is proposed. The type strain is MX6T (=IBRC M 10928T=LMG 28548T).

Salinifilum gen. nov., with description of Salinifilum proteinilyticum sp. nov., an extremely halophilic actinomycete isolated from Meighan wetland, Iran, and reclassification of Saccharopolyspora aidingensis as Salinifilum aidingensis comb. nov. and Saccharopolyspora ghardaiensis as Salinifilum ghardaiensis comb. nov.

A Gram-positive, halophilic actinobacterial strain Miq-12T was isolated from Meighan wetland in Iran. Strain Miq-12T was strictly aerobic, catalase positive and oxidase negative. The isolate grew at 12-25 % NaCl, at 30-50 °C and pH 5.5-10.5. The optimum NaCl, temperature and pH for growth were 15-20 %, 40 °C and 7.0-8.0, respectively. The cell wall of strain Miq-12T contained meso-diaminopimelic acid as diagnostic diamino acid and arabinose as whole-cell sugar. The polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine and phosphatidylinositol. It synthesized cellular fatty acids of anteiso and iso-branched types, anteiso-C17 : 0, iso-C17:0, iso-C15:0, iso-C16 : 0. The major respiratory quinone was MK-9(H4). The G+C content of its genomic DNA was 72.1 mol%. Phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that strain Miq-12T belongs to the family Pseudonocardiaceae, constituted a separate clade, and showed the closest phylogenetic similarity to Saccharopolyspora aidingensis TRM 46074T (96.99 %) and Saccharopolyspora ghardaiensis CCUG 63370T (96.92 %). On the basis of phylogenetic analysis, phenotypic and chemotaxonomic characteristics, a novel genus and species of the family Pseudonocardiaceae, Salinifilum proteinilyticum gen. nov., sp. nov., are proposed. The type strain is Miq-12T (=IBRCM 11033T=LMG 28390T). We also propose that S. aidingensis and S. ghardaiensis should be transferred to this new genus and be named Salinifilum aidingensis comb. nov. and Salinifilum ghardaiensis comb. nov., respectively. The type strain of Salinifilum aidingensis comb. nov. is TRM 46074T (=CCTCCAA 2012014T=JCM 30185T) and the type strain of Salinifilum ghardaiensis comb. nov. is CCUG 63370T (=DSM 45606T=CECT 8304T).

Prauserella oleivorans sp. nov., a halophilic and thermotolerant crude-oil-degrading actinobacterium isolated from an oil-contaminated mud pit.

A crude-oil-degrading, Gram-stain-positive actinobacterial strain, RIPIT, was isolated from a soil sample collected from an oil-contaminated mud pit in Khangiran oil and gas field, in the north-east of Iran. RIPIT was strictly aerobic, catalase- and oxidase-positive. The strain grew with 0-12.5 % (w/v) NaCl (optimum 3-5 %), at 25-55 °C (optimum 45 °C) and at pH 6.0-9.5 (optimum pH 7.0). The results of 16S rRNA gene sequence comparative analysis indicated that RIPIT represents a member of the genus Prauserella, with high phylogenetic similarity to Prauserella coralliicola SCSIO 11529T (97.5 %), Prauserella endophytica SP28S-3T (97.5 %) and Prauserella marina MS498T (97.2 %). DNA-DNA relatedness values between the novel strain and P. coralliicola DSM 45821T, P. endophytica DSM 46655T and P. marina DSM 45268T were 28 , 19 and 23 %, respectively. The cell wall peptidoglycan of RIPIT contained meso-diaminopimelic acid as the diamino acid and the whole-cell sugars are galactose and arabinose. The polar lipids pattern contained phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol and two unknown phospholipids. Its cellular fatty acids pattern consisted of C17 : 1ω6c, iso-C16 : 0 and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH), and the major respiratory quinone was MK-9(H4). The G+C content of the genomic DNA was 69 mol%. On the basis of polyphasic taxonomic data we propose that RIPIT represents a novel species of the genus Prauserella, for which the name Prauserella oleivorans sp. nov. is proposed. The type strain of Prauserellaoleivorans is RIPIT (=IBRC-M 10906T=LMG 28389T).

Soortia roseihalophila gen. nov., sp. nov., a new taxon in the order Balneolales isolated from a travertine spring, and description of Soortiaceae fam. nov.

A novel Gram-stain-negative, slightly halophilic, motile, curved rod with a horseshoe shape, designated strain Bsw-2bT, was isolated from Badab-Soort travertine spring in Iran. Phylogenetic analysis of the 16S rRNA gene sequence revealed that strain Bsw-2bT belongs to the order Balneolales, showing 84.6 % sequence similarity to Gracilimonastropica DSM 19535T and 84.4 % and 83.9 % sequence similarity to Gracilimonas rosea CL-KR2T and Balneola vulgaris DSM 17893T, respectively. In addition, phenotypic and physiological features could clearly differentiate strain Bsw-2bT from species of the most closely related genera, Gracilimonas, Balneola, Aliifodinibius and Fodinibius. The strain was able to grow with 1-3 % (w/v) (optimum at 2 %) NaCl, at temperatures of 28-34 °C (optimum at 30 °C) and between pH 6.0 and 8.0 (optimum at pH 7.0). The major cellular fatty acids of strain Bsw-2bT were iso-C15 : 0, iso-C13 : 0 and iso-C14 : 0. The polar lipid profile of strain Bsw-2bT was composed predominantly of phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, an unknown glycolipid and four unknown phospholipids. The DNA G+C content was 40.5 mol%. Based on the evidence from the polyphasic study, strain Bsw-2bT represents a novel species in a novel genus within a new family, for which the name Soortia roseihalophila gen. nov., sp. nov. is proposed, within the new family Soortiaceae fam. nov. The type strain is strain Bsw-2bT (=IBRC-M 10915T=LMG 28547T).

Nocardia halotolerans sp. nov., a halotolerant actinomycete isolated from saline soil.

A novel halotolerant actinomycete, strain Chem15(T), was isolated from soil around Inche-Broun hypersaline wetland; its taxonomic position was determined based on a polyphasic approach. Strain Chem15(T) was strictly aerobic and tolerated NaCl up to 12.5%. The optimum temperature and pH for growth were 28-30 °C and pH 7.0-7.5, respectively. The cell wall of strain Chem15(T) contained meso-diaminopimelic acid as diamino acid and galactose, arabinose and ribose as whole-cell sugars. The major phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannosides. The cellular fatty acids profile consisted of C16 : 0, iso-C18 : 0, C18 : 0 10-methyl and C18 : 1ω9c, and the major respiratory quinone was MK-8(H4cycl). The G+C content of the genomic DNA was 68.0 mol%. The novel strain constituted a distinct phyletic line within the genus Nocardia, based on 16S rRNA gene sequence analysis, and was closely associated with Nocardia sungurluensis DSM 45714(T) and Nocardia alba DSM 44684(T) (98.2 and 98.1% 16S rRNA gene sequence similarity, respectively). However DNA-DNA relatedness and phenotypic data demonstrated that strain Chem15(T) was clearly different from closely related species of the genus Nocardia. It is concluded that the organism should be classified as a representative of a novel species of the genus Nocardia, for which the name Nocardia halotolerans sp. nov. is proposed. The type strain is Chem15(T) ( = IBRC-M 10490(T) = LMG 28544(T)).

Salininema proteolyticum gen. nov., sp. nov., a halophilic rare actinomycete isolated from wetland soil, and emended description of the family Glycomycetaceae.

A Gram-stain-positive actinobacterial strain, Miq-4T, was isolated from soil around Meighan wetland in the centre of Iran. Strain Miq-4T was strictly aerobic, catalase- and oxidase-positive. The isolate grew in the presence of 3­15 % (w/v) NaCl, at 20­40 °C and pH 6.0­11.0. The optimum NaCl, temperature and pH for growth were 7.0 %, 30 °C and 7.0­8.5, respectively. The cell wall of strain Miq-4T contained meso-diaminopimelic acid as the diamino acid and glucose and ribose as the whole-cell sugars. The polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. Strain Miq-4T synthesized cellular fatty acids of anteiso- and iso-branched types, including anteiso-C17 : 0, anteiso- C15 : 0 and iso-C16 : 0, and the major respiratory quinone was MK-9(H4). The G+C content of the genomic DNA was 68.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences and characteristic patterns of 16S rRNA gene signature nucleotides revealed that strain Miq-4T belongs to the family Glycomycetaceae and showed the closest phylogenetic similarity with Haloglycomyces albus YIM 92370T (94.1 % 16S rRNA gene sequence similarity). On the basis of phylogenetic analysis and phenotypic and chemotaxonomic characteristics, strain Miq-4T represents a novel species of a new genus in the family Glycomycetaceae, for which the name Salininema proteoliyticum gen. nov., sp. nov. is proposed. The type strain of the type species is Miq-4T ( = IBRC-M 10908T = LMG 28391T). An emended description of the family Glycomycetaceae is also proposed in order to include features of the new genus.

Alloactinosynnema iranicum sp. nov., a rare actinomycete isolated from a hypersaline wetland, and emended description of the genus Alloactinosynnema.

A Gram-staining-positive actinobacterial strain, Chem10(T), was isolated from soil around Inche-Broun hypersaline wetland in the north of Iran. Strain Chem10(T) was strictly aerobic, and catalase- and oxidase-positive. The isolate grew with 0-3 % NaCl, at 20-40 °C and at pH 6.0-8.0. The optimum temperature and pH for growth were 30 °C and pH 7.0, respectively. The cell wall of strain Chem10(T) contained meso-diaminopimelic acid as diamino acid and galactose, ribose and arabinose as whole-cell sugars. The polar lipid pattern contained diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Strain Chem10(T) synthesized cellular fatty acids of the straight-chain saturated and mono-unsaturated, and iso- and anteiso-branched types C14 : 0, C16 : 0, iso-C16 : 1, anteiso-C17 : 0, iso-C16 : 0, iso-C14 : 0 and iso-C15 : 0, and the major respiratory quinone was MK-9(H4). The G+C content of the genomic DNA was 70.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain Chem10(T) belonged to the family Pseudonocardiaceae and showed the closest phylogenetic similarity to Alloactinosynnema album KCTC 19294(T) (98.3 %) and Actinokineospora cibodasensis DSM 45658(T) (97.9 %). DNA-DNA relatedness values between the novel strain and strains Alloactinosynnema album KCTC 19294(T) and Actinokineospora cibodasensis DSM 45658(T) were only 52 % and 23 %, respectively. On the basis of phylogenetic analysis, phenotypic characteristics and DNA-DNA hybridization data, a novel species of the genus Alloactinosynnema is proposed, Alloactinosynnema iranicum sp. nov. The type strain is Chem10(T) ( = IBRC-M 10403(T) = CECT 8209(T)). In addition, an emended description of the genus Alloactinosynnema is proposed.