Partial 16S rDNA Sequencing Research Articles

The antibacterial activity and silk dyeing of the crude pigment extract from J4 actinobacteria strain

Actinobacteria represent a vast variety of prokaryotic microbes that produce a diverse group of secondary metabolites used in medical, agricultural, and industrial applications. The objectives in this study were to investigate the antibacterial activities and potential for silk dyeing of the crude extract from the actinobacterial strain J4. Initially, the antibacterial activities of the crude pigment extracts obtained from the actinobacterial strain J4 isolated from the wasp’s nest were determined. To extract the water-insoluble pigments used for antibacterial activity testing via the agar well diffusion method, ethyl acetate was applied during the pigment extraction. These extracts showed an inhibitory effect against Salmonella Typhimurium TISTR1472, Bacillus subtilis PSRU-01, and Enterococcus faecalis TISTR459. Secondly, evaluation of silk dyeing was carried out. We used the extractant as 70 % (v/v) ethanol in the alternative extraction process when the strain J4 was cultivated by solid-state fermentation with broken-milled rice to prepare actinobacterial dye for silk dyeing. The pigment extract obtained was tested for color stability based on washing methods, and the sensitivity to light was determined according to ISO105-C06:2010(E) and ISO 105-B02:2014(E), respectively. The color stability and sensitivity of the pigment extract to washing showed scores between 3-5, and the sensitivity to light demonstrated scores between 3-4. Taxonomically, the strain J4 was dark-violet colonies, and its water-soluble pigment diffused on sodium caseinate agar (SCA). To identify the strain J4, the phylogenetic identification based on partial sequencing of 16S rDNA was applied. The identification result indicated that the strain J4 was closely related to Streptomyces coacervatus (99.8% sequence similarity). In future, antibacterial activities of the crude extracts obtained from actinobacteria strain J4 should be tested with other food-borne pathogens. The development of Streptomyces dyes should also focus on the stability of the dyes during the silk dyeing process.

A new species of slender flatworm in the genus Eucestoplana and a record of E. cf. cuneata (Platyhelminthes, Polycladida) from the Okinawa Islands, Japan, with an inference of their phylogenetic positions within Cestoplanidae

In this study, we describe a new species of elongated marine flatworm, Eucestoplana ittanmomensp. nov., collected from the intertidal zone of the Okinawa Islands, Japan. Eucestoplana ittanmomensp. nov. is distinguished from other congeners based on the following characteristics: i) its translucent body lacking coloration, ii) its dome-shaped penis sheath, iii) the absence of cilia on the inner wall of the male atrium except outside the penis sheath, and iv) the presence of an adhesive organ at the posterior end of the body. Additionally, we report the occurrence of E. cf. cuneata (Sopott-Ehlers & Schmidt, 1975) in Japan; E. cuneata has previously been documented in the Galapagos and Fiji Islands. We conducted phylogenetic analyses to infer the positions of the two Eucestoplana species within Cestoplanidae using a concatenated dataset comprising partial 18S and 28S rDNA sequences from E. cf. cuneata and E. ittanmomensp. nov. from Japan, as well as four known Cestoplana species with sequences available in public databases. Our phylogenetic analyses revealed that Cestoplana and Eucestoplana were reciprocally monophyletic. Furthermore, the genetic distance of the 16S rDNA sequences supported the genetic independence of the two sister species, E. cf. cuneata and E. ittanmomensp. nov.

Species of Ergasilus von Nordmann, 1832 (Copepoda: Ergasilidae) from cichlid fishes in Lake Tanganyika.

Ergasilus (von Nordmann, 1832) (Ergasilidae) is a species-rich group of parasitic copepods with a wide distribution in freshwater, marine and brackish environments. Up to now, 9 species of Ergasilus are known from cichlid fishes in Africa. In this study, 5 species, including 3 new, were collected from the gills of 12 cichlid species (11 genera: Bathybates, Ctenochromis, Eretmodus, Gnathochromis, Lamprologus, Neolamprologus, Ophthalmotilapia, Perissodus, Simochromis, Spathodus and Tanganicodus) of the northeastern shore of Lake Tanganyika in Burundi, namely E. macrodactylus (Sars, 1909), E. megacheir (Sars, 1909), E. caparti n. sp., E. parasarsi n. sp. and E. parvus n. sp. All species found were identified and described on the basis of adult female specimens using an integrative taxonomy approach mixing morphological characterization and molecular analyses of 2 ribosomal DNA markers (partial 18S and 28S rDNA sequences). An identification key for Ergasilus species from Lake Tanganyika is included. This study provides the first molecular data for Ergasilus species in Africa. The phylogenetic analyses suggest that the Ergasilus species parasitizing Lake Tanganyikan cichlids form a well-supported clade within the Ergasilidae. However, their phylogenetic relationships with other congeners still remain unclear due to a lack of molecular data for this diverse genus.

Isolation and characterization of Priestia megaterium KD7 for the biological control of pear fire blight.

Erwinia amylovora is a plant pathogen that causes fire blight disease in Rosaceous plants, such as pear and apple. To develop an effective biocontrol method to suppress E. amylovora, a total of 16 bacteria were isolated from pear orchard soil in China and screened for antagonistic activity in vitro. Among them, 9 isolates that exhibited antagonistic activity against E. amylovora were identified, including Bacillus atrophaeus, Priestia megaterium (previously known as Bacillus megaterium) and Serratia marcescens based on the partial 16S rDNA sequence analysis and similarity search. The plate confrontation experiments showed that strain 8 (P. megaterium strain KD7) had strong antagonistic activity against E. amylovora. The methanolic extract from cell-free supernatant of strain KD7 displayed high antibacterial activities against E. amylovora. Furthermore, the active compounds of strain KD7 were separated by thin layer chromatography (TLC) and the amino acids were detected by the presence of a spot with retention factor (Rf) of 0.71. Next, three lipopeptides were identified with high-resolution mass spectrometry (HRMS), including C13-surfactin [M+H]+ at m/z 1008.14, C15-surfactin [M+H]+ at m/z 1036.50, and C14-iturin A [M+H]+ at m/z 1043.17. Strain KD7 showed multiple antibiotic resistance, such as ampicillin, erythromycin, penicillin and tetracycline. The detached pear leaves, twigs and fruits assay showed that both protective and curative action with strain KD7 had the ability to decrease the development of fire blight. Taken together, P. megaterium strain KD7 is a potential effective biocontrol agent against fire blight.

Morphology and molecular phylogeny of Chauhanellus Bychowsky & Nagibina, 1969 (Monogenoidea) parasitizing marine catfish (Ariidae) from the Atlantic coast of South America: a new species, supplementary taxonomic information and new insights.

In this study, a new dactylogyrid species is described from the gills of two ariid fish species of the Brazilian coast, Genidens barbus and Genidens genidens, by combining morphological characters with partial 18S rDNA sequences. The new species can be distinguished from its congeners by the following morphological characteristics: a male copulatory organ (MCO), a coiled tubular shaft of around one counterclockwise ring, base with a sclerotized cap; MCO with tapered distal region; a T-shaped accessory piece; a vagina, a cup-shaped vaginal vestibule, slightly sclerotized; and a pharynx comprising a muscular, glandular bulb. Supplementary taxonomic data for Chauhanellus velum from Sciades couma (type-host) are also presented and new 18S rDNA sequences of Chauhanellus spp. from other ariid fish from South America are provided. Phylogenetic analyses based on partial 18S rDNA gene sequences placed Chauhanellus riograndinensis n. sp. and Chauhanellus velum as two early divergent lineages within Chauhanellus from South America. Finally, a way to test the monophyly of Chauhanellus and Hamatopeduncularia is also discussed, which may be useful for future studies.

Characterization of a yellow pigmented, alkali and heavy metal tolerant Glutamicibacter sp. isolated from red mud

Red mud ponds are waste dumping sites of bauxite mines where highly alkaline and polluted red mud or bauxite residues are deposited. They harbour many microbial strains of environmental importance. In the present study, a yellow coloured bacterial colony was isolated which showed tolerance to high pH, and metals like lead, chromium and arsenic. Genomic DNA extraction followed by partial 16S rDNA amplification and sequencing study revealed the isolate to be Glutamicibacter sp. The bacterial strain showed luxuriant growth in pH ranging from pH5 to pH 11, and salt concentration up to 8% NaCl. The pigment was isolated by methanol extraction and identified to be beta-carotene through FTIR spectroscopy. The pigment showed absorption peaks at 436 nm and 466 nm. The pigment could not show antibacterial activity significantly. The pigments showed a good free radical scavenging activity which increased significantly under stress conditions. No such studies were done from the red mud ponds of Utkal Alumina earlier. Tolerance of the bacterial strain to various heavy metals indicates presence of heavy metals in the red mud. Identification of molecular mechanisms underlying the heavy metal resistance and use of the pigments as cosmetics can be carried out in future.

Oswaldocruzia franciscoensis n. sp. (Nematoda: Molineidae) in Leptodactylus macrosternum Miranda-Ribeiro, 1926 (Anura: Leptodactylidae) from Caatinga morphoclimatic domain, Brazil: morphological and molecular characterisation.

A new species of Oswaldocruzia Travassos, 1917 (Nematoda, Molineidae), parasite of Leptodactylus macrosternum Miranda-Ribeiro, 1926 (Anura: Leptodactylidae), from Caatinga morphoclimatic domain, Brazil, is described based on morphological and molecular data. Oswaldocruzia franciscoensis n. sp. is characterised by an anterior extremity with a cephalic vesicle divided into two portions, a body covered by cuticular longitudinal ridges, and cervical alae. Males of the new species have caudal bursa of type I with a 2-1-2 pattern, spicules divided into a shoe, bifurcated fork, and blade with two unequal branches, in which the longer branch bifurcates at its distal portion end and the smaller branch with three distal processes, each with distal bifurcations. Females have didelphic and amphidelphic uteri, an ovijector divided into vestibule, anterior and posterior sphincters, and anterior and posterior infundibula. The new species differs from its Neotropical congeners that have caudal bursa of type I, based on the presence of cervical alae and by having a spicular blade distally divided into two unequal branches, with the longer branch bifurcating at its distal portion and smaller branch with three distal processes, each distally bifurcated. The partial 18S rDNA sequence generated for Oswaldocruzia franciscoensis n. sp. is the first of a representative belonging to this genus in the Neotropical region.

Characterization of Five Psychrotolerant Alcanivorax spp. Strains Isolated from Antarctica.

Five psychrotolerant Alcanivorax spp. strains were isolated from Antarctic coastal waters. Strains were screened for molecular and physiological properties and analyzed regarding their growth capacity. Partial 16S rDNA, alk-B1, and P450 gene sequencing was performed. Biolog EcoPlates and the API 20E test were used to evaluate metabolic and biochemical profiles. Bacterial growth in sodium acetate was determined at 4, 15, 20, and 25 °C to evaluate the optimal temperature. Furthermore, the ability of each strain to grow in a hydrocarbon mixture at 4 and 25 °C was assayed. Biosurfactant production tests (drop-collapse and oil spreading) and emulsification activity tests (E24) were also performed. Concerning results of partial gene sequencing (16S rDNA, alk-B1, and P450), a high similarity of the isolates with the same genes isolated from other Alcanivorax spp. strains was observed. The metabolic profiles obtained by Biolog assays showed no significant differences in the isolates compared to the Alcanivorax borkumensis wild type. The results of biodegradative tests showed their capability to grow at different temperatures. All strains showed biosurfactant production and emulsification activity. Our findings underline the importance to proceed in the isolation and characterization of Antarctic hydrocarbon-degrading bacterial strains since their biotechnological and environmental applications could be useful even for pollution remediation in polar areas.

Optimization of Degenerate PCR Conditions for Reducing Error Rates in Detection of PKS and NRPS Gene groups in Actinomycetes.

The screen of Polyketide Synthase (PKS) and Nonribosomal Peptide Synthetase (NRPS) gene groups is a quick way to discover new therapeutic agents. However, errors in laboratory techniques cause a loss of touch with reality. This study aimed to evaluate the presence of PKS and NRPS gene groups in previously isolated strains by optimizing their specialized amplification by degenerate primers and indicating the evolutionary relationships with reference strains. PKS-I, II, and NRPS genes PCR amplification was performed using three degenerate primer sets for 22 actinomycete strains with antibacterial activity. Annealing temperature and the amount of template DNA and primers were optimized. PCR products of PKS-I, II, and NRPS from three strains were sequenced after TA cloning. Besides, strains with high antibacterial activity were identified by biochemical features and partial 16S rDNA sequencing and hypothetically classified by a phylogenetic tree. High frequencies of PKS-I (86.4%), PKS-II (81.8%), and NRPS (95.4%) genes were found among the strains after optimization. Fourteen strains (64%) contained all of the genes, and 100% of strains had at least two genes. These numbers are pretty distinct in comparison with the previous researches. All of the sequenced strains were members of Streptomyces genus. Our research showed that degenerate PCR strongly depends on the variation of annealing temperature and primer concentration, resulting in an unexpected shift in PCR outputs. The sequencing results confirmed the optimized conditions for specialized PCR of PKS-I, PKS-II, and NRPS gene groups.

Probiotic Potentials of Lactic Acid Bacteria and Yeasts from Raw Goat Milk in Nigeria.

Probiotic microorganisms are incorporated in foods due to their numerous health benefits. We investigated lactic acid bacteria (LAB) and yeasts isolated from goat milk in Nigeria for novel probiotic strains. In this study, a total of 27 LAB and 23 yeast strains were assessed for their probiotic potentials. Only six LAB strains (Weissella cibaria GM 93m3, Weissella confusa GM 92m1, Pediococcus acidilactici GM 18a, Pediococcus pentosaceus GM 23d, Lactiplantibacillus pentosus GM 102s4, Limosilactobacillus fermentum GM 30m1) and four yeast strains (Candida tropicalis 12a, C. tropicalis 33d, Diutina rugosa 53b, and D. rugosa 77a) identified using partial 16S and 26S rDNA sequencing, respectively, showed survival at pH 2.5, 0.3% bile salt, and simulated gastrointestinal conditions and possessed auto-aggregative and hydrophobic properties, thus satisfying key in vitro criteria as probiotics. All LAB strains showed coaggregation properties and antimicrobial activities against pathogens. Pediococcus pentosaceus GM 23d recorded the strongest coaggregation percentage (34-94%) against 14 pathogens, while W. cibaria GM 93m3 showed the least (6-57%) against eight of the 14 pathogens. The whole cell and extracellular extracts of LAB and yeast strains, with the exception of D. rugosa 77a, had either 2,2-diphenyl-1-picryl-hydrazyl and/or hydroxyl radical scavenging activity. In conclusion, all six LAB and four yeast strains are important probiotic candidates that can be further investigated for use as functional starter cultures.

Antagonistic potential of an Egyptian entomopathogenic nematode, compost and two native endophytic bacteria isolates against the root-knot nematode (Meloidogyne incognita) infecting potato under field conditions

BackgroundThe root-knot nematode, Meloidogyne spp., are one of the most dominant and dangerous group of pests. The deformations and discolorations make tubers unmarketable and/or of less quality. Therefore, management of Meloidogyne spp. becomes an obligatory challenge that warrants intervention. Biological control agents are the best alternative tools for controlling plant-parasitic nematodes that comply with the requirements of the development of the green agriculture and that reduce the reliance on these harmful chemicals. Therefore, this study aimed to evaluate the effectiveness of compost singly, and in combinations with the bio-agents Heterorhabditis bacteriophora, and two bacterial isolates Nem 212 and Nem 213 against the root-knot nematode Meloidogyne incognita infecting potato plants under field conditions.ResultsAmong 15 bacterial isolates (Nem205-Nem219) obtained from the rhizosphere of tomato and eggplant from Giza, Egypt, the two isolates (Nem 212 and Nem 213) were molecularly characterized based on the partial 16S rDNA sequencing analysis. These two bacterial isolates were deposited in the GenBank as Bacillus cereus Nem 212 and B. cereus Nem 213 and were tested against M. incognita J2s in vitro. Results showed that the cell filtrates of B. cereus Nem 212 and B. cereus Nem 213 gave the highest percentage of M. incognita J2s mortality (100%), after 48 h of the in vitro application. Moreover, all the applied treatments significantly suppressed the reproductive of M. incognita on potato plants and enhanced the potato crop yield under the field conditions. Compost enriched with B. cereus Nem 212 cell suspension was the most effective treatment. The combination between the bacterial cell suspension and the compost offered an increase in the disease curing and the potato plant growth and yield production, compared to the treatment with compost alone. The entomopathogenic nematode, Heterorhabditis bacteriophora, was relatively less effective in controlling M. incognita on potato, compared to B. cereus Nem 212 and/or B. cereus Nem 213 treatments. However, when compost was enriched with H. bacteriophora, it increased its capability to control the nematodes.ConclusionsThis study provides insights into the practical usage of EPNs H. bacteriophora, and the endophytic bacteria (B. cereus Nem 212 or B. cereus Nem 213) as biocontrol agents against M. incognita on potato plants. The application of compost enriched with the bacterial cell suspensions of either B. cereus Nem 212 or B. cereus Nem 213 and H. bacteriophora within Galleria mellonella cadaver proved efficient control of M. incognita infecting potato plants and improved the growth and yield of potato plants under field conditions.

Polyphasic study of phytopathogenic bacterial strains associated with deep bark canker of walnut in Serbia revealed a new species, Brenneria izbisi sp. nov.

Serious outbreaks of walnut deep bark canker were observed on young walnut trees (Juglans regia L.) in two localities in the northern part of Serbia during 2020. From the symptomatic walnut tissues, two types of bacterial colonies were isolated, predominantly, light cream, circular and smooth colonies, as well as small, yellowish, mucoid and convex ones. PCR analysis and phenotypic assays suggested that the former group belongs to Brenneria spp., while the latter isolates were identified as Xanthomonas arboricola pv. juglandis. Within the Brenneria group, two strains were identified as Brenneria nigrifluens, while other 15 strains did not belong to any Brenneria species described so far. Therefore, we selected four representative strains of the unknown Brenneria sp. and subjected them to polyphasic analysis. As expected, in a phylogenetic tree based on partial 16S rDNA sequences, four novel strains grouped with other Brenneria representatives, and showed close phylogenetic relationship to Brenneria salicis. Furthermore, multilocus sequence analysis (MLSA) based on the partial sequences of atpD, gyrB, infB and rpoB housekeeping genes and core-genome phylogeny indicated that the studied strains form a novel and a clearly separate Brenneria lineage. Overall genome relatedness indices showed that they represent a new Brenneria species. The new species can be differentiated from the other Brenneria spp. infecting walnut and closely related B. salicis strains based on phenotypic characteristics, as well. Moreover, the pathogenicity tests on two-year-old walnut plants proved the ability of strains to cause necrosis and longitudinal black lesions and cracks on the trunk and branches of walnut trees. Overall, polyphasic characterization showed that the studied strains isolated from walnut with symptoms of deep bark canker represent a novel species of the genus Brenneria for which the name Brenneria izbisi sp. nov. is proposed. The type strain of B. izbisi is KBI 423T (= CFBP 9035T = LMG 32479T). To facilitate rapid identification of newly described species, a conventional PCR protocol and primers targeting the putative gene hrpP, were developed. Further study should reveal the potential role of each pathogen isolated from symptomatic walnut in disease development as well as possible interaction between them.

Reversible shifts between interstitial and epibenthic habitats in evolutionary history: Molecular phylogeny of the marine flatworm family Boniniidae (Platyhelminthes: Polycladida: Cotylea) with descriptions of two new species.

Tiny animals in various metazoan phyla inhabit the interstices between sand and/or gravel grains, and adaptive traits in their body plan, such as simplification and size reduction, have attracted research attention. Several possible explanations of how such animals colonized interstitial habitats have been proposed, but their adaptation to this environment has generally been regarded as irreversible. However, the actual evolutionary transitions are not well understood in almost all taxa. In the present study, we show reversible evolutionary shifts from interstitial to epibenthic habitats in the lineage of the polyclad flatworm genus Boninia. In addition, we establish two new species of this genus found from different microhabitats on a single beach in Okinawa Island, Japan: (i) the interstitial species Boninia uru sp. nov. from gravelly sediments and (ii) the epibenthic species Boninia yambarensis sp. nov. from rock undersurfaces. Our observations suggest that rigid microhabitat segregation exists between these two species. Molecular phylogenetic analyses based on the partial 18S and 28S rDNA sequences of the new Boninia species and four other congeners, for which molecular sequences were available in public databases [Boninia antillara (epibenthic), Boninia divae (epibenthic), Boninia neotethydis (interstitial), and an unidentified Boninia sp. (habitat indeterminate)], revealed that the two interstitial species (B. neotethydis and B. uru sp. nov.) were not monophyletic among the three epibenthic species. According to ancestral state reconstruction analysis, the last common ancestor of the analyzed Boninia species inhabited interstitial realms, and a shift to the epibenthic environment occurred at least once. Such an "interstitial to noninterstitial" evolutionary route seems to be rare among Animalia; to date, it has been reported only in acochlidian slugs in the clade Hedylopsacea. Our phylogenetic tree also showed that the sympatric B. uru sp. nov. and B. yambarensis sp. nov. were not in a sister relationship, indicating that they colonized the same beach independently rather than descended in situ from a common ancestor that migrated and settled at the beach.

Exposure to bacterial and fungal bioaerosols in facilities processing biodegradable waste.

The aim of the study was to determine the exposure of workers within biodegradable waste processing facilities to bacteria and fungi to identify any exposures of potential concern to health. Occupational measurements were performed in six composting and three bioenergy (bioethanol or methane/biogas) producing facilities. Bioaerosols were measured from breathing zones with Button aerosol or open face cassette filter samplers, and swab specimens were taken from the nasal mucous membranes of the workers. Aspergillus fumigatus, Bacillus cereus group, Campylobacter spp., Salmonella spp., Streptomyces spp., and Yersinia spp. were determined by real-time polymerase chain reaction (qPCR). A. fumigatus, and mesophilic and thermophilic actinobacteria were also cultivated from filters. Bacterial airborne endotoxins collected by IOM samplers were analyzed using a Limulus assay. Bioaerosol levels were high, especially in composting compared to bioenergy producing facilities. Endotoxin concentrations in composting often exceeded the occupational exposure value of 90 EU/m3, which may be harmful to the health. In addition to endotoxins, the concentrations of A. fumigatus (up to 2.4 × 105 copies/m3) and actinobacteria/Streptomyces spp. (up to 1.6 × 106 copies/m3) in the air of composting facilities were often high. Microbial and endotoxin concentrations were typically highest in waste reception and pre-treatment, equal or decreased during processing and handling of treated waste, and lowest in wheel loader cabins and control rooms/outdoors. Still, the parameters measured in wheel loader cabins were often higher than in the control sites, which suggests that the use of preventive measures could be improved. B. cereus group, Salmonella spp., and Yersinia spp. were rarely detected in bioaerosols or nasal swabs. Although Campylobacter spp. DNA was rarely detected in air, as a new finding, Campylobacter ureolyticus DNA was frequently detected in the nasal mucous membranes of workers, based on partial 16S rDNA sequencing. Moreover, especially A. fumigatus and C. ureolyticus spp. DNA concentrations in swabs after the work shift were significantly higher than before the shift, which indicates their inhalation or growth during the work shift. Microbial qPCR analysis of bioaerosols and swab samples of nasal mucosa allowed measuring exposure in various work operations and during the work shift, identifying problems for health risk assessment to improve working conditions, and evaluating the effectiveness of preventive measures and personal protection of workers.

Efficient role of rice associated Plant growth promoting bacteria

Rice-associated plant growth-promoting bacteria (PGPB) have a major role in promoting growth anddevelopment using a variety of mechanisms. In this study, we isolated bacteria from drought stressedfields to check their effects on rice plants. Bacterial strains were selectively isolated based on their growthon selective media and different biochemical tests. These bacteria were identified by partial sequencing of16s rDNA. We have found that two isolates, Brevibacterium sp. (B4) and Priestiaaryabhattai (B16) (MZ081422.1),have a magnificent role in the growth of rice plants compared to mock-treated plants after drought stresstreatment.

Characterization of halo-tolerant plant growth promoting endophytic Bacillus licheniformis MHN 12

BackgroundEndophytic bacteria overlay significant role in plant growth promotion, eliminating phyto-pathogens and combating stress-conditions. In the present study, we aimed to screen high salt tolerant bacteria and study their adaptive response to elevated salt concentrations. A total of 46 endophytic bacterial isolates from Vigna radiata were screened for salt tolerance. The high salt tolerant endophytic isolate was characterized for alteration in morphology, growth rate, protein profiling, and compatible solute concentrations. ResultsThe isolate MHN12, based upon biochemical characterization and partial 16S rDNA sequencing identified as B. licheniformis (accession number MG273753) was able to tolerate up to 15% NaCl (Sodium Chloride) (2.6 M) concentration. The isolate possessed 1-aminocyclopropane-1-carboxylic acid deaminase (ACCD) activity along with indole acetic acid (IAA), siderophore, ammonia, organic acid and hydrogen cyanide (HCN) production. Accumulation of proline was apparent up to 7.5% NaCl concentration and declined afterwards. Ultrastructure analysis using TEM (transmission electron microscopy) revealed the morphological alteration from rods to filaments. ConclusionAcclimatization to salt stress and plant growth promoting activities could contribute to utilization of this bacterium as bioinoculant to enhance the crop yield and discourage the application of chemical fertilizers.

Phylogenetic relationships of Ochoterenatrema Caballero, 1943 (Digenea: Lecithodendriidae) with descriptions of two new species.

Ochoterenatrema is a small genus of lecithodendriid digeneans that includes six species parasitic in Neotropical bats in North and South America. Members of this genus are characterized by the presence of a pseudogonotyl formed by thickened tegument on the left side of the ventral sucker. We examined morphology of specimens belonging to five species of Ochoterenatrema from Brazil, Ecuador and the USA and describe two new Ochoterenatrema spp. (Ochoterenatrema sphaerula n. sp. and Ochoterenatrema gracilis n. sp.). These new species are readily differentiated from their congeners by a combination of morphological characters, including the distribution of vitelline follicles and the body shape. We have provided the first DNA sequences for Ochoterenatrema. Newly generated partial nuclear 28S rDNA and mitochondrial cox1 gene sequences were used to differentiate among species and study the phylogenetic affinities of Ochoterenatrema spp., including one of the new species, O. gracilis n. sp., as well as Ochoterenatrema diminutum, Ochoterenatrema fraternum and Ochoterenatrema cf. labda. The phylogeny of the Microphalloidea based on 28S had well-supported topology, particularly at the family level. The Ochoterenatrema clade was strongly supported; however, the internal topology of the clade was weakly supported. Comparison of sequences revealed 0.4-1.3% interspecific divergence in 28S and 9.1-19.7% interspecific divergence in cox1 among Ochoterenatrema spp. We hypothesize that extremely diverse fauna of bats in South and Central America harbors multiple undescribed species of Ochoterenatrema. Several lecithodendriid genera from bats and other hosts are yet to be included in future molecular phylogenetic analyses to test the monophyly of this extremely diverse digenean family and analyze evolutionary affinities of its constituent taxa.

Genetic distance and phylogenetic relationships of some Echinostoma species (E. malayanum, E. revolutum, E. miyagawai) and Hypoderaeum conoideum (family Echinostomatidae) inferred from partial 28S rDNA sequence analysis

Echinostomiasis is a neglected disease caused by the intestinal flukes (family Echinostomatidae, suborder Echinostomata) and is common in communities in Asian countries, such as India, Indonesia, the Philippines, China, Malaysia, Singapore, Korea, Japan, Thailand, Laos, Cambodia and Vietnam. The genetic markers from the nuclear ribosomal transcription units are commonly used in genetic studies and phylogenetic analyses. A portion of the 28S rDNA sequence (domains D1–D3, of 1062–1067 bp for the final use) was obtained from the zoonotic Echinostoma malayanum (strain E/Amala-EMI3-TH), E. revolutum (strain Erevo-MSD15-TH), E. miyagawai (Emiya-RED11-TH), and Hypoderaeum conoideum (Hcono-RED42-TH) species; and used to perform an alignment for genetic distance estimation and phylogenetic analysis. The alignment was performed using 62 strains of 42 species from 19 genera of the family Echinostomatidae, including Echinoparyphium, Echinostoma, Artyfechinostomum, Patagifer, Neoacanthoparyphium, Hypoderaeum, Echinoparyphium, Drepanocephalus, Euparyphium, Chaunocephalus, Neopetasiger, Ribeiroia, Cathaemasia, Rhopalias, Isthmiophora, Petasiger, Moliniella, Pegosomum, and Schistosoma (family Schistosomatidae). The genetic distance estimation among 16 strains/10 species has shown a low intra-specific divergence level between strains within the same species, such as E. miyagawai (0–0.10%), E. revolutum (0.10–0.50%), and H. conoideum (0–0.10%), while between strains within the same genus it was higher (normally over 1.0%) and among strains/species between genera it was the highest (3.06–4.12%). The 28S rDNA sequence analysis and phylogenetic relationship well supported the Echinostoma/ Artyfechinostomum malayanum intergeneric taxonomy and the topology indicated clear, well-supported positions of member species in different genera in the family Echinostomatidae of the suborder Echinostomata. More sensu lato samples of the genera, are required for sequencing, particularly those of zoonotic species in the five genera: Artyfechinostomum, Echinostoma, Hypoderaeum, Echinoparyphium, and Isthmiophora. The resultant mitochondrial and nuclear data obtained from these species will be a good source to use to clearly assess the taxonomic and generic relationships.

Bacillus aryabhattai SMNCH17-07 Strain: First Isolation and Characterization from Textile Waste Water with Evaluation of Its Decolorization Ability against Azo Dyes

Azo dyes are largely entered into various industrial products. Conventional treatment techniques are insufficient to eliminate azo dyes entirely from effluents due to the ability of these dyes to maintain color, steadiness and being difficult to transformation. Bioremediation procedures have -been proved as a promising method to remove azo dyes effectively, due to their advantages such as the low cost, being environmentally safe and the large scale usage capability. 
 In this study, SA3 bacterial isolate, has been collected from textile waste water area in industrial zone located in Usak province, Turkey. Phenotypic identification and phylogenetic determination on the basis of partial 16S rDNA sequence comparisons indicated that this strain is 100 % Bacillus aryabhattai SMNCH17-07 strain. SA3 isolate was Gram positive, central spore-forming and rod-shape bacteria .The isolate gave positive results for catalase, NaCl, Voges-Proskauer, starch hydrolysis assays and negative results for the citrate reaction.
 The decolorizing ability of this strain was evaluated against CI Acid Blue 193 CI 15707 and CI Acid Red 88 CI 15620 dyes. The results showed that absorbance rate of Acid Blue by this isolate was (33, 17%) after 216 hours. While Absorbance rate of Acid red by this isolate was (62,68 %) after120 hours. According to FTIR spectrometer results of dyes adsorption it was found that bacterial retention sites were possibly aromatic and aliphatic (C = C, C = N, N = N) as well C‐O groups. As the first report on the isolation of Bacillus aryabhattai SMNCH17-07 strain from textile waste water with the evaluation of its ability to remove azo dyes, we suggest testing this bacterium as a low cost and ecofriendly bioremediator agent against further harmful dyes and pollutants.

Plant growth-promoting bacteria isolated from sugarcane improve the survival of micropropagated plants during acclimatisation

The plant microbiome plays an important role in nutrient acquisition and buffering plant hosts against abiotic and biotic stress. During in vitro propagation of sugarcane, pathogenic microorganisms are eliminated and most of the beneficial endophytic microorganisms. The objective of this study was to isolate and characterise potential plant growth-promoting bacteria (PGPB) from sugarcane and to analyse their ability to improve the survival of micropropagated sugarcane plantlets during the acclimatisation stage. First, bacterial isolates from sugarcane were identified by partial 16S rDNA sequencing and tested for plant growth-promoting (PGP) features, such as inorganic and organic phosphate solubilisation nitrogen fixation, siderophore synthesis, indole-3-acetic acid production, tolerance to abiotic stress and antibiotics production. Then three bacterial strains with multiple PGP traits were independently applied to micropropagated seedlings of the sugarcane variety TUC 03-12 when the plants were transferred to a nursery for ex vitro acclimatisation. The effect of selected PGPB on survival rates of micropropagated plantlets was evaluated in three independent assays, using different batches of seedlings. Thirty days after inoculation, 182-Bacillus and 336-Pseudomonas isolates significantly improved the transferred plants survival rate. High variability in plant survival among independent experiments was observed, but treatments with the 336-Pseudomonas strain showed a low mortality rate (20%) in all assays. This procedure constitutes a biological tool to improve the survival of micropropagated plants during greenhouse acclimatisation. Furthermore, it provides an initial tool for selecting bacteria with possible PGP effects in the field. Highlights - A total of 162 isolates obtained from the rhizosphere, rhizoplane, roots, and stems of sugarcane were characterised for plant growthpromoting features and identified by partial 16S rDNA sequencing. - Two PGPBs strains isolated from sugarcane (182-Bacillus and 336-Pseudomonas) significantly improved survival rates of micropropagated seedlings during the acclimatisation stage. - Under different stress conditions, the 336-Pseudomonas strain improved the survival of micropropagated plants during the acclimatisation stage.