Fatty Acid Methyl Ester Analysis Research Articles

Fatty Acid Profiles and Fuel Properties of Oils from Castor Oil Plants Irrigated by Microalga-treated Wastewater

Microalgae can function as a bio-fertilizer and a bio-filter for wastewater treatment, thus allowing the re-use of wastewater for the irrigation of plants that produce abundant bio-oils and for the irrigation of low-quality soils. Chlorella vulgaris was grown in Om El-Resh drain water as growth medium, after enrichment with nitrogen and phosphorous. Castor seeds were cultivated in a greenhouse at the Algal Biotechnology Unit, National Research Centre, Egypt. Fifteen days later, transplants were irrigated by untreated wastewater (WW); wastewater treated with microalgae (WW+A); or wastewater treated with microalgae followed by removal of microalgae (WW-A). Oil extraction was performed by seed warming and grinding with n-hexane, followed by soaking, filtration, and passage through Silica gel 60. Esterification was performed, and then fatty acid methyl esters (FAMEs) were determined. The results indicated that the wastewater treatment markedly affected seed oil content. The WW+A treatment led to the highest seed oil yield (41.8%), followed WW-A treatment (28.14%), and the WW treatment (25.12%). FAME analysis indicated that the presented fatty acids of castor oil were C16 and C18 and ricinoleic acid (C18:1) was the most abundant (83.1 to 84.63%). In spite of the higher seed oil content when plants were grown in WW+A, there were differences in the fuel proprieties of seed oils in the different groups, based on American Society for Testing and Materials criteria. Our results suggest that wastewater can be successfully used for irrigation of soils that have poor fertility to produce bio-oils during land reclamation.

Tannerella serpentiformis sp. nov., isolated from the human mouth.

Three strains representing the previously uncultured human oral Tannerella taxon HMT-286 were recently isolated from the subgingival plaque of a patient with chronic periodontitis. The phenotypic and genetic features of strain SP18_26T were compared to those of the type species of Tannerella, Tannerella forsythia. A genome size of 2.97 Mbp (G+C content 56.5 mol%) was previously reported for SP18_26T, compared to a size of 3.28 Mbp (47.1 mol%) in T. forsythia ATCC 43037T. 16S rRNA gene sequence comparisons also revealed 94.3 % sequence identity with T. forsythia ATCC 43037T. Growth was stimulated by supplementation of media with N-acetyl muramic acid, as seen with T. forsythia, but the cells displayed a distinctive snake-like morphology. Fatty acid methyl ester analysis revealed a profile differing from T. forsythia, chiefly in the amount of 3-OH-16 : 0 (four-fold lower in SP18_26T). Overall, metabolic enzyme activity also differed from T. forsythia, with enzyme activity for indole present, but the complement of glycoside hydrolase enzyme activity was smaller than T. forsythia, for example, lacking sialidase and N-acetyl-β-glucosaminidase - evidence backed up by analysis of its gene content. On the basis of these results, a new species Tannerella serpentiformis is proposed for which the type strain is SP18_26T (=DSM 102894T=JCM 31303T).

Identification and validation of the mode of action of the chalcone anti-mycobacterial compounds

ObjectivesThe search for new TB drugs has become one of the great challenges for modern medicinal chemistry. An improvement in the outcomes of TB chemotherapy can be achieved by the development of new, shorter, cheap, safe and effective anti-TB regimens. MethodsChalcones (1a-1o) were synthesized and evaluated for their antimycobacterial activity against Mycobacterium bovis BCG using growth inhibition assays. Compound 1a was selected as a ‘hit’ compound. The mode of action of compound 1a, was identified by mycolic acid methyl esters (MAMEs) and fatty acid methyl esters (FAMEs) analysis using thin layer chromatography. Dose dependent experiments were conducted by over-expressing components of FAS-II in M. bovis BCG to confirm the target. Ligand binding using intrinsic tryptophan assay and molecular docking were used to further validate the target. ResultsMAMEs and FAMEs analysis showed dose-dependent reduction of MAMEs with the overall abundance of FAMEs suggesting that compound 1a targets mycolic acid biosynthesis. Direct binding of 1a to InhA was observed using an intrinsic tryptophan fluorescence binding assay, and a 2-fold IC50 shift was observed with an InhA overexpressing strain confirming InhA as the cellular target. ConclusionThe chalcone 1a exhibits potent antimycobacterial activity, displays a good safety profile and is a direct inhibitor of InhA, a key component in mycolic acid synthesis, validating this series for further anti-TB drug development.

Oleaginous yeasts isolated from traditional fermented foods and beverages of Manipur and Mizoram, India, as a potent source of microbial lipids for biodiesel production

PurposeOleaginous yeasts can accumulate intracellular lipid bodies or triacylglycerides (TAGs) under nutrient limiting conditions. TAGs derived from those yeast strains are considered as an alternative to conventional plant-based oils for biodiesel production. In this study, we attempt to isolate and characterize yeast strains from selected traditional fermented foods of Manipur and Mizoram, India, and study their oleaginous attributes for biodiesel production.MethodFourteen potential oleaginous yeasts were isolated from fermented food samples of Manipur and Mizoram, India. The isolates were identified by 5.8S internal transcribed spacer (ITS) rRNA gene sequencing. Intracellular TAG accumulation by yeast cells were confirmed by Nile red fluorescence microscopy and spectrometry technique. The most promising isolates were evaluated for lipid accumulation having different initial carbon to nitrogen (C/N) ratios and also the full kinetic studies (depicting the glucose consumption, biomass, and lipid production) using optimum C/N ratio were estimated. Fatty acid methyl esters (FAME) profile of the transesterified lipids were analyzed by GC-MS.ResultsThe identified yeast isolates belonged to seven different genera viz. Rhodotorula, Pichia, Candida, Saturnispora, Wickerhamomyces, Zygoascus, and Saccharomyces. Under nitrogen-limiting conditions, maximum biomass concentration of 5.66 ± 0.03 g/L and 4.713 ± 0.03 g/L was produced by Wickerhamomyces anomalus FK09 and Pichia kudriavzevii FK02, respectively. The highest lipid concentration (g lipid/L fermentation broth) of 0.58 g/L was attained by Rhodotorula mucilaginosa R2, followed by Wickerhamomyces anomalus FK09 (0.51 g/L), and Zygoascus hellenicus FC10 (0.41 g/L). Rhodotorula mucilaginosa R2 exhibited the maximum lipid content (% lipid/g dry cell weight) of (21.63 ± 0.1%) after 96 h of growth. The C/N ratio of 40 and 20 was found to be optimum for R. mucilaginosa R2 and W. anomalus FK09 with a lipid content of 22.21 ± 0.4% and 12.83 ± 0.08% respectively.ConclusionNewly isolated yeast strains were obtained from traditional fermented food samples of Manipur and Mizoram, India. FAME analysis of the transesterified lipid extracts suggested the potential use of yeast-derived oil as an alternative to vegetable oil for biodiesel production.

Highly accurate classification of biological spores by culture medium for forensic attribution using multiple chemical signature types and machine learning.

Future proliferation of biological expertise and new technology may increasingly lower the difficulty to produce biological organisms for misuse. Rapid attribution of a biological attack is needed to quickly identify the person or lab responsible and prevent additional attacks by enabling the apprehension of suspects. Here, triplicate batches of Bacillus anthracis Sterne strain (BaSt) spores were grown in a total of seven amateur and professional media. Multiple orthogonal analytical signatures (peptides, metabolites, lipids by fatty acid methyl ester (FAME) analysis, bulk organic profile, and trace elements) were collected from the BaSt spores. The proteomics and metabolomics analyses identified promising attribution signature compounds that are unique to each of the seven production methods. In addition, while each of the signature types showed varying degrees of value individually for attributing BaSt spores to the culture medium used to prepare them, fusing results from all five signatures types to increase sourcing robustness and using a random forest sourcing algorithm yielded 100% hold-one-batch-out cross-validation classification accuracy and an average relative source probability for the correct source 5.5× higher than the most probable incorrect source. These preliminary results provide a proof-of-concept for the development of forensic examinations that can attribute biological agents to production methods for use in future investigations.

Proposal of Lactobacillus kosoi Chiou et al. 2018 as a later heterotypic synonym of Lactobacillus micheneri McFrederick et al. 2018, elevation of Lactobacillus plantarum subsp. argentoratensis to the species level as Lactobacillus argentoratensis sp. nov., and Lactobacillus zhaodongensis sp. nov., isolated from traditional Chinese pickle and the intestinal tract of a honey bee (Apis mellifera).

Lactobacillus kosoi Chiou et al. 2018 and Lactobacillus micheneri McFrederick et al. 2018 are closely related, and they share 100 % 16S rRNA gene sequence similarity, 99.6 % pheS gene sequence similarity, 100 % rpoA gene sequence similarity, 97.3 % average nucleotide identity (ANI) value and 76.6 % in silico DNA-DNA hybridization (isDDH) value, indicating that they represent the same species. Fatty acid methyl esters (FAME) analysis and phenotypic characterization also indicated that L. kosoi and L. micheneri are very similar. We propose L. kosoi Chiou et al. 2018 as a later heterotypic synonym of L. micheneri McFrederick et al. 2018. The taxonomic position of Lactobacillus plantarum subsp. argentoratensis in the L. plantarum group was re-examined using a polyphasic approach, including sequence analyses of 16S rRNA, pheS, rpoA and recA genes, average nucleotide identity analysis, in silico DNA-DNA hybridization, fatty acid methyl ester analysis and phenotypic characterization. Results of 16S rRNA gene sequence analysis indicated that L. plantarum subsp. argentoratensis was closely related to L. plantarum subsp. plantarum, L. pentosus and L. paraplantarum in the L. plantarum group, sharing 99.6-99.7 % 16S rRNA gene sequence similarities. Results of pheS, rpoA and recA gene sequence analyses indicated that L. plantarum subsp. argentoratensis was most closely related to L. plantarum subsp. plantarum, having 91.8 % pheS gene sequence similarity, 98.9 % rpoA gene sequence similarity and 93.1 % recA gene sequence similarity. L. plantarum subsp. argentoratensis DSM 16365T shared 95.6 % ANI value and 62.9 % isDDH value with L. plantarum subsp. plantarum ATCC 14917T. The low isDDH value confirmed that L. plantarum subsp. argentoratensis and L. plantarum subsp. plantarum represent two different species, rather than two different subspecies in the L. plantarum group. On the basis of the data from polyphasic characterization obtained in the present study and in previous studies, L. plantarum subsp. argentoratensis is elevated to the species level and represents a novel species of the genus Lactobacillus, for which the name Lactobacillus argentoratensis sp. nov. is proposed and the type strain is DKO 22T (=CIP 108320T=DSM 16365T=JCM 16169T). Two novel Gram-stain-positive bacterial strains, designated 1206-1T and F027-1-2, were isolated from traditional pickle in Heilongjiang Province, PR China, and from the intestinal tract of a honey bee (Apis mellifera) in Hubei Province, PR China, respectively. The two bacteria were characterized by a polyphasic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, fatty acid methyl ester analysis, average nucleotide identity analysis, in silico DNA-DNA hybridization analysis and an analysis of phenotypic features. The results of 16S rRNA gene sequence analysis indicated that strains 1206-1T and F027-1-2 were distantly related to Lactobacillus sharpeae, Lactobacillus hulanensis, Lactobacillus songhuajiangensis, Lactobacillus pantheris, Lactobacillus thailandensis, Lactobacillus camelliae, Lactobacillus jixianensis, Lactobacillus nasuensis, Lactobacillus baoqingensis, Lactobacillus manihotivorans and Lactobacillus porcinae. Strain 1206-1T exhibited 94.2-96.4 % 16S rRNA gene sequence similarities, 69.5-83.3 % pheS gene sequence similarities and 73.1-90.3 % rpoA gene sequence similarities to type strains of phylogenetically related species. ANI and isDDH values between strain 1206-1T and the type strains of phylogenetically related species were 52.7-73.7 % and 21.1-30.1 %, respectively. On the basis of the data obtained in the present study, a novel species, Lactobacillus zhaodongensis sp. nov. is proposed and the type strain is 1206-1T (=CCM 8981T=CCTCC AB 2019200T=LMG 31620T).

Identification and culturing of cyanobacteria isolated from freshwater bodies of Sri Lanka for biodiesel production.

The present study was carried out to investigate cyanobacteria as a potential source for biodiesel production isolated from fresh water bodies of Sri Lanka. Semi mass culturing and mass culturing were carried out to obtain biomass for extracting total lipids. Fatty acid methyl ester (FAME) or biodiesel was produced from extracted lipid by trans-esterification reaction. FAME component was identified using gas chromatography (GC). Atotal of 74 uni-algal cultures were obtained from Biofuel and Bioenergy laboratory of the National Institute of Fundamental Studies (NIFS), Kandy, Sri Lanka. The total lipid content was recorded highest in Oscillatoria sp. (31.9 ± 2.01% of dry biomass) followed by Synechococcus sp. (30.6 ± 2.87%), Croococcidiopsis sp. (22.7 ± 1.36%), Leptolyngbya sp. (21.15 ± 1.99%), Limnothrixsp. (20.73 ± 3.26%), Calothrix sp. (18.15 ± 4.11%) and Nostoc sp. (15.43 ± 3.89%), Cephalothrixsp. (13.95 ± 4.27%), Cephalothrix Komarekiana (13.8 ± 3.56%) and Westiellopsisprolifica (12.80 ± 1.97%). FAME analysis showed cyanobacteria contain Methyl palmitoleate, Linolelaidic acid methyl ester, Cis-8,11,14-eicosatrienoic acid methyl ester, Cis-10-heptadecanoic acid methyl ester, Methyl myristate, Methyl pentadecanoate, Methyl octanoate, Methyl decanoate, Methyl laurate, Methyl tridecanoate, Methyl palmitoleate, Methyl pentadeconoate, Methyl heptadeconoate, Linolaidic acid methyl ester, Methyl erucate, Methyl myristate, Myristoloeic acid, Methyl palmitate, Cis-9-oleic acid methyl ester, Methyl arachidate and Cis-8,11,14-ecosatrieconoic acid methyl ester. The present study revealed that cyanobacteria isolated from Sri Lanka are potential source for biodiesel industry because of their high fatty acid content. Further studies are required to optimize the mass culture conditions to increase thelipid content from cyanobacterial biomass along with the research in the value addition to the remaining biomass.

Lactobacillus hegangensis sp. nov., Lactobacillus suibinensis sp. nov., Lactobacillus daqingensis sp. nov., Lactobacillus yichunensis sp. nov., Lactobacillus mulanensis sp. nov., Lactobacillus achengensis sp. nov., Lactobacillus wuchangensis sp. nov., Lactobacillus gannanensis sp. nov., Lactobacillus binensis sp. nov. and Lactobacillus angrenensis sp. nov., isolated from Chinese traditional pickle and yogurt.

Fourteen Gram-stain-positive bacterial strains were isolated from Chinese traditional pickle and yogurt. The strains were characterized using a polyphasic taxonomic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, fatty acid methyl ester analysis, determination of DNA G+C content, determination of average nucleotide identity (ANI), in silico DNA-DNA hybridization (isDDH) and an analysis of phenotypic features. The data demonstrated that the 14 strains represented ten novel species belonging to the genus Lactobacillus, strains 73-4T, 247-3T, 143-4(a)T, 33-1T, 143-6T, 247-4T, 17-4T, 143-1T, 735-2T and M1530-1T were designated as the type strains. Strains 73-4T and 247-3T were phylogenetically related to the type strains of Lactobacillus camelliae and Lactobacillus jixianensis, having 97.0-98.9 % 16S rRNA gene sequence similarities, 83.9-87.2 % pheS gene sequence similarities and 86.8-93.3 % rpoA gene sequence similarities. Strains 143-4(a)T and 33-1T were phylogenetically related to the type strains of Lactobacillus rhamnosus, Lactobacillus paracasei and Lactobacillus casei, having 93.6-96.5 % 16S rRNA gene sequence similarities, 73.9-77.2 % pheS gene sequence similarities and 76.1-77.6 % rpoA gene sequence similarities. Strains 143-6T, 247-4T, 17-4T and 143-1T were phylogenetically related to the type strains of Lactobacillus concavus, Lactobacillus dextrinicus and Lactobacillus bayanensis, exhibiting 95.5-99.9 % 16S rRNA gene sequence similarities, 76.5-83.1 % pheS gene sequence similarities and 83.6-98.3 % rpoA gene sequence similarities. Strain 735-2T was phylogenetically related to the type strains of Lactobacillus zhaoyuanensis, Lactobacillus jiayinensis and Lactobacillus coryniformis, having 98.2-99.1 % 16S rRNA gene sequence similarities, 82.8-84.1 % pheS gene sequence similarities and 93.0-93.9 % rpoA gene sequence similarities. Strain M1530-1T was phylogenetically related to the type strains of Lactobacillus suantsaiihabitans and Lactobacillus brevis, having 99.5 and 99.0 % 16S rRNA gene sequence similarities, 90.3 and 81.7 % pheS gene sequence similarities and 97.7 and 91.1 % rpoA gene sequence similarities. The ANI and isDDH values between strains 73-4T, 247-3T, 143-4(a)T, 33-1T, 143-6T, 247-4T, 17-4T, 143-1T, 735-2T, M1530-1T and type strains of phylogenetically related species were less than 86.8 % and 33.9 % respectively, confirming that they represent ten novel species within the genus Lactobacillus. Based upon the data of polyphasic characterization obtained in the present study, ten novel species, Lactobacillus hegangensis sp. nov., Lactobacillus suibinensis sp. nov., Lactobacillus daqingensis sp. nov., Lactobacillus yichunensis sp. nov., Lactobacillus mulanensis sp. nov., Lactobacillus achengensis sp. nov., Lactobacillus wuchangensis sp. nov., Lactobacillus gannanensis sp. nov., Lactobacillus binensis sp. nov. and Lactobacillus angrenensis sp. nov., are proposed and the type strains are 73-4T (=NCIMB 15177T=CCM 8912T=CCTCC AB 2018407T), 247-3T (=NCIMB 15176T=JCM 33275T), 143-4(a)T (=NCIMB 15173T=CCM 8948T=JCM 33273T=CCTCC AB 2018390T), 33-1T (=NCIMB 15169T=CCM 8947T=JCM 33272T=CCTCC AB 2018405T), 143-6T (=NCIMB 15162T=CCM 8951T=JCM 33274T=CCTCC AB 2018411T), 247-4T (=NCIMB 15155T=CCM 8897T=LMG 31059T=CCTCC AB 2018410T), 17-4T (=NCIMB 15161T=CCM 8946T=JCM 33271T=CCTCC AB 2018406T), 143-1T (=NCIMB 15157T=CCM 8937T=CCTCC AB 2018409T), 735-2T (=NCIMB 15190T=CCM 8925T=LMG 31186T) and M1530-1T (=NCIMB 15150T=CCM 8893T=LMG 31046T=CCTCC AB 2018402T), respectively.

Weissella sagaensis sp. nov., isolated from traditional Chinese yogurt.

Three Gram-stain-positive bacterial strains, designated X0750T, X0278 and X0401, isolated from traditional yogurt in Tibet Autonomous Region, PR China, were characterized by a polyphasic approach, including sequence analyses of the 16S rRNA gene and three housekeeping genes (pheS, rpoA and recA), determination of average nucleotide identity (ANI) and average amino acid identity (AAI), in silico DNA-DNA hybridization (isDDH), fatty acid methyl ester (FAME) analysis and phenotypic characterization. Strain X0750T was phylogenetically related to the type strains of Weissella hellenica, Weissella bombi, Weissella paramesenteroides, Weissella jogaejeotgali, Weissella thailandensis, Weissella oryzae, Weissella cibaria and Weissella confusa, having 94.4-100 % 16S rRNA gene sequence similarities, 76.7-90.0 % pheS gene sequence similarities, 88.9-99.4 % rpoA gene sequence similarities and 77.6-92.8 % recA gene sequence similarities, respectively. ANI, isDDH and AAI values between strain X0750T and type strains of phylogenetically related species were less than 90.4, 40.9 and 92.8 % respectively, confirming that strain X0750T represents a novel species within the genus Weissella. Based upon the data obtained in the present study, a novel species, Weissella sagaensis sp. nov., is proposed and the type strain is X0750T(=NCIMB 15192T=CCM 8924T=LMG 31184T=CCTCC AB 2018403T).

Evaluation of the progeny produced by interspecific hybridization between Camelina sativa and C. microcarpa

Camelina (Camelina sativa, Brassicaceae) has attracted interest in recent years as a novel oilseed crop, and an increasing number of studies have sought to enhance camelina's yield potential or to modify the composition of its oil. The ability of camelina to cross-hybridize with its wild relative, C. microcarpa, is of interest as a potential source of genetic variability for the crop. Manual crosses were performed between the crop C. sativa and its wild relative C. microcarpa; F1 and F2 progenies were obtained. Cytology was used to study meiosis in the parents and F1s and to evaluate pollen viability. Flow cytometry was used to estimate nuclear DNA amounts and fatty acid methyl ester analysis was used to evaluate the lipid composition of F3 seeds. The F1 plants obtained by interspecific crossing presented severe abnormalities at meiosis and low pollen viability, and produced very few F2 seeds. The F2s presented diverse phenotypes and in some cases severe developmental abnormalities. Many F2s were aneuploid. The F2s produced highly variable numbers of F3 seeds, and certain F3 seeds presented atypical lipid profiles. Considering the meiotic abnormalities observed and the probability of aneuploidy in the F2 plants, the C. microcarpa accessions used in this study would be difficult to use as sources of genetic variability for the crop.

Evaluation of performance and validity limits of gas chromatography electron ionisation with Orbitrap detection for fatty acid methyl ester analyses.

While the GC-Orbitrap, marketed in 2015, represents a technological breakthrough in terms of sensitivity, resolution and mass stability, many studies have reported ion ratio modification in mass spectra using the standard 70 eV electron ionisation. We studied the influence of the acquisition and sample parameters leading to these modifications on fatty acid methyl esters (FAMEs). FAMEs showed that these variations in relative intensities of ions were related to the acquisition parameters such as the mass range and the offset values of the C-TRAP, but also directly related to the column concentration of the sample, and especially that it was molecule-dependent. Advantageously, it is possible to use this feature to promote the molecular ions of FAMEs sometimes not present in a spectrum under electron ionisation at 70 eV. The 70 eV electron ionisation mass spectra from the GC-Orbitrap were clearly molecule-dependent and could be due to metastable ions during storage states in the C-TRAP.

Bacillus miscanthi sp. nov., a alkaliphilic bacterium from the rhizosphere of Miscanthus sacchariflorus.

A novel bacterial strain, designated AK13T (=KACC 21401T=DSM 109981T), was isolated from the rhizosphere of Miscanthus sacchariflorus. Strain AK13T was found to be an aerobic, Gram-stain-positive, endospore-forming and rod-shaped bacterium. It formed yellow circular colonies with smooth convex surfaces. The genomic DNA G+C content of strain AK13T was estimated to be 40 mol%. Phylogenetic analysis based on 16S rRNA gene sequence similarity showed that this strain was most closely related to Bacillus lehensis MLB2T (99.4 %), Bacillus oshimensis K11T (98.8 %) and Bacillus patagoniensis PAT 05T (96.6 %). The average nucleotide identity values between strain AK13T and B. lehensis MLB2T, B. oshimensis K11T and B. patagoniensis PAT 05T were 90.93, 91.05 and 71.87 %, respectively, with the digital DNA-DNA hybridization values of 42.7, 42.6 and 18.8 %, respectively. Cells grew at 5-40 °C (optimum, 28-35 °C), pH 6.5-13 (optimum, pH 8-9) and in the presence of 0-13.0 % (w/v) NaCl (optimum, 1 %). The cell wall of strain AK13T contained meso-diaminopimelic acid, and the major isoprenoid quinone was MK-7. Results of fatty acid methyl ester analysis revealed that iso-C15 : 0 was the predominant cellular fatty acid. Two-dimensional thin-layer chromatography analysis indicated that the major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and glycolipid. The genotypic and phenotypic characteristics suggested that strain AK13T represented a novel species of the genus Bacillus, and thus the name Bacillus miscanthi sp. nov. is proposed.

Characteristic of Fatty Acids Biotransform from Hermetia Illucens Prepupae Fed with Various Organic Wastes Before Conversion to Methyl Ester Form

Farming of Black solder fly (Hermetia illucens) had received great research interest by scientist to investigate the application of this insect’s larvae as a means of organic waste biotransformation, feed for aquaculture and husbandry as well as for biodiesel feedstock. The compositions of this insect’s larvae is well known to have high content of lipid and protein, this has demonstrates its potential use as animal feed and alternative energy. To the best of our knowledge, this study provides the ultimate investigation on the characteristic of the pre-pupae fatty acids than that of the feeds before conversion to methyl ester for biodiesel production. The study also investigates the impact of different feeds on fatty acid profile of Hermetia illucens. The farming and breeding of Hermetia illucens fly lifecycle was set up and the populations were maintained at Universiti Tunku Abdul Rahman (UTAR), Kampar campus. The newly hatched neonates were fed on fruit waste and food waste, respectively. Continuous feeding was dispensed to the neonates until it became pre-pupae. At this stage, cessations of feeding and self-harvesting by the pre-pupae were observed. The harvesting of the pre-pupae was done at this instar. Thereafter, the harvested live pre-pupae were cleaned and processed into dried pre-pupae. The dried pre-pupae were kept in an air tight container for further use. The characteristic of the feed and the dried pre-pupae were determined in a series of analysis such as crude lipid extraction, free fatty acid, acid value and fatty acid methyl ester analysis. All the analysis was done in accordance to AOAC methods and each analysis was repeated in triplicates. The results from the study shows that, Hermetia illucens pre-pupae crude lipids content as well as fatty acid profile was influenced by types of feeds as their diet. The lipid content found in the larvae biomass were profound regardless the types of feed. Comparatively, the amount of crude lipid obtained from Hermetia illucens pre-pupae was found to be higher than that of all feeds. The amount of crude lipids extracted from feeds of fruit waste and food waste were approximately 1.24 ± 0.06 and 3.27 ± 0.28 wt%, respectively. The amount of pre-pupae’s crude lipid derived from fruit waste and food waste were higher by 35.85 and 11.48 times, respectively. It is noteworthy that, increase in the lipid accumulation in larvae biomass indicates that the larvae are able to assimilate and bio-convert the feed composition into their own biomass. Consequently, the nutrient composition contains in the types of feed, ingested and digested by the larvae strongly affect the overall lipid accumulation. Moreover, the fatty acid extracted from the Hermetia illucens biomass showed approximately 1.77 times higher in saturated fatty acid (SFA) than that of fruit waste and food waste. Lauric acid is the predominant saturated fatty acid found in the prepupae. The SFA attained by the pre-pupae derived from fruit waste and food wastes were 84.7 and 81.5 wt%, respectively. While the total unsaturated fatty acids were found to be higher in the feeds than that of the pre-pupae. Meanwhile, the total amount of unsaturated fatty acid from fruit waste and fruit waste were attained at 50.9 and 53.7 wt%, respectively. In conclusion, the next stage of this study will be carried out to evaluate the biodiesel quality of the converted fatty acid methyl ester derived from Hermetia illucens fatty acid.

Fatty acid methyl ester analysis of Aspergillus fumigatus isolated from fruit pulps for biodiesel production using GC-MS spectrometry

ABSTRACT In this study, a total of six fungal samples were isolated from apple, strawberry and orange pulp. DNA sequence analysis was used as molecular identification method. ITS region was aligned in DNA sequence analysis, and an algorithm sequence similarity was done using BLAST (Basic Local Alignment Search Tool) program to identify these isolates. All the six isolates were identified as Aspergillus fumigates. The total lipid content was varied in the isolates which were ranged from 29.4 to 21.0 mg/100 ml. Moreover, the obtained lipid form mycelium biomass of the isolates was transesterified by a base catalyst. The methyl esters were analyzed by using GC-MS. GC-MS Spectrometry revealed the presence of different fatty acids with long chain (C11:0, C15:0, C17:1, C18:2, C16:1). High efficiency biodiesel can be obtained using long-chain fatty acids. Fatty acid profiles of A. fumigatus isolated from different fruit pulps have confirmed its potentiality as well as showed the beneficial utilization of these fatty acids for biodiesel production.

COST EFFECTIVE CULTIVATION AND BIOMASS PRODUCTION OF GREEN MICROALGA DESMODESMUS SUBSPICATUS MB. 23 IN NPK FERTILIZER MEDIUM

Microalgal biomass has emerged as a promising alternative to replace plant-based biofuel feedstock due to its higher lipid productivity. But cultivation of microalgae in expensive analytical grade culture media is a major obstacle in feasible algal biofuel production. Hence the present investigation was carried out to find an alternative, low-cost culture medium for the increased biomass yield and biochemical production of microalga, Desmodesmus subspicatus MB. 23. The strain was cultivated in different concentrations of 19: 19: 19 NPK media and checked for the algal biomass production and biochemical accumulation. Maximum algal cell density (5290 ×104 cells/ ml), biomass yield and productivity (2.72 g/L, 60.87 mg/L/d) was attained in 2 g/ L NPK fertilizer medium, whereas 1g/L medium exhibited increased chlorophyll production (chl-a, 4.7 mg/g d wt, chl-b 1.7 mg/g d wt). High level of carotenoid accumulation (4.6 mg/g d wt) as well as lipid accumulation (29.5%) was found in 0.5g/L fertilizer media and Fatty acid methyl ester analysis of the strain showed abundant production of C16 and C18 fatty acids. Findings of the current study proved that, lower concentration of 19:19:19 NPK fertilizer can be used for the enhanced production of green microalgae. Other than cost effective biofuel production, NPK fertilizer grown D. subspicatus can also be used for the production of a wide range of metabolites such as food and feed additives, pharmaceuticals and cosmetics.

Apibacter muscae sp. nov., a novel bacterial species isolated from house flies.

We describe the isolation and characterization of three bacterial isolates from the common house fly, Musca domestica, caught in Londerzeel, Belgium and Huye District, Rwanda. Although isolated from distinct geographical locations, the strains show >99 % identical 16S rRNA gene sequences and are <95 % identical to type strains of Apibacter species. Whole-genome sequences were obtained for all three strains. The genomes are 2.4-2.5 Mb with a G+C content of ~30.3 mol%. Bacteriological and biochemical analysis of the strains demonstrate distinctly different characteristics compared to known Apibacter species. Particularly, the three strains investigated in this study can be distinguished from the known Apibacter species (Apibacter mensalisand Apibacter adventoris) through urease and β-glucosidase activities. Whole-cell fatty acid methyl ester analysis shows that the fatty acid composition of the novel strains is also unique. On the basis of phylogenetic, genotypic and phenotypic data, we propose to classify these isolates as representatives of a novel species of the genus Apibacter, Apibacter muscae sp. nov., in reference to its prevalence in house flies, with strain G8T (=LMG 30898T=DSM 107922T) as the type strain.

Effect of Light Intensity on Algal Growth in Designed Hybrid Photobioreactor and Biodiesel Production

In current scenario algal fuel is considered as the most viable solution for the depleting non-renewable fuel sources. This study demonstrates the controlled, continuous production of the microalgae species Chlorella Pyrenoidosa in a novel designed hybrid photobioreactor aimed to the later production of biodiesel. The reactor is operated at different light intensity and its effect on the biomass production and pH was assessed. The final biomass is harvested by centrifugation preceded by sedimentation from which the algal lipids are extracted using solvent extraction method and biodiesel by acid catalysed transesterification reaction is produced. The species population density is found to be exponential at 7130 LUX at a photoperiod of 8/16(L/D). The biomass growth in the developed design is established as a factor of 2.88 with respect to optical density. At the light intensity of 7130LUX, the developed Hybrid Photobioreactor yields a dry algal mass of 2.09g/L with a lipid productivity of 1.01gL-1day-1 and 70.62% of fatty acid methyl esters (FAME). The analysis of FAME is done using gas chromatography and a maximum of 73.4% biodiesel composition is observed.

Lactobacillus yilanensis sp. nov., Lactobacillus bayanensis sp. nov., Lactobacillus keshanensis sp. nov., Lactobacillus kedongensis sp. nov., Lactobacillus baiquanensis sp. nov., Lactobacillus jidongensis sp. nov., Lactobacillus hulinensis sp. nov., Lactobacillus mishanensis sp. nov. and Lactobacillus zhongbaensis sp. nov., isolated from Chinese traditional pickle and yogurt.

Fourteen Gram-stain-positive bacterial strains were isolated from Chinese traditional pickle and yogurt. The strains were characterised using a polyphasic taxonomic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, determination of DNA G+C content, determination of average nucleotide identity, fatty acid methyl ester analysis and an analysis of phenotypic features. The data demonstrated that the fourteen strains represented nine novel species belonging to the genus Lactobacillus, strains 54-2T, 54-5T, 33-7T, 116-2T, 184-8T, 204-8T, 8-1(1)T, 256-3T and M1575T were designated as the type strains. Strain 54-2T was phylogenetically related to the type strains of Lactobacillus composti and Lactobacillus floricola, having 96.5 and 91.6 % 16S rRNA gene sequence similarities, less than 74.6 % pheS gene sequence similarities, less than 81.6 % rpoA gene sequence similarities and less than 72.5 % ANI values. Strain 54-5T was phylogenetically related to the type strains of Lactobacillus dextrinicus and Lactobacillus concavus, exhibiting 99.1 and 97.3 % 16S rRNA gene sequence similarities, less than 83.1 % pheS gene sequence similarities, less than 93.1 % rpoA gene sequence similarities and less than 79.9 % ANI values. Strains 33-7T, 116-2T, 184-8T, 204-8T, 8-1(1)T, 256-3T and M1575T were phylogenetically related to the type strains of Lactobacillus tucceti, Lactobacillus nodensis, Lactobacillus insicii, Lactobacillus allii, Lactobacillus metriopterae, Lactobacillus terrae, Lactobacillus versmoldensis and Lactobacillus furfuricola, sharing 95.6-100 % 16S rRNA gene sequence similarities, less than 91.6 % pheS gene sequence similarities, less than 98.2 % rpoA gene sequence similarities and less than 89.4 % ANI values. Based upon the data of polyphasic characterisation obtained in the present study, nine novel species, Lactobacillus yilanensis sp. nov., Lactobacillus bayanensis sp. nov., Lactobacillus keshanensis sp. nov., Lactobacillus kedongensis sp. nov., Lactobacillus baiquanensis sp. nov., Lactobacillus jidongensis sp. nov., Lactobacillus hulinensis sp. nov., Lactobacillus mishanensis sp. nov. and Lactobacillus zhongbaensis sp. nov., are proposed and the type strains are 54-2T (=NCIMB 15154T=CCM 8896T=KCTC 21120T=LMG 31058T), 54-5T (=NCIMB 15151T=CCM 8894T), 33-7T (=NCIMB 15153T=CCM 8936T=KCTC 21118T=LMG 31166T), 116-2T (=NCIMB 15158T=CCM 8899T=KCTC 21124T=LMG 31051T), 184-8T (=NCIMB 15152T=CCM 8895T=KCTC 21131T=LMG 31050T), 204-8T (=NCIMB 15159T=CCM 8900T=KCTC 21133T=LMG 31054T), 8-1(1)T (=NCIMB 15156T=CCM 8898T=KCTC 21115T=LMG 31047T), 256-3T (=NCIMB 15160T=CCM 8901T=LMG 31048T) and M1575T (=NCIMB 15149T=CCM 8892T=LMG 31045T), respectively.

Lactobacillus pingfangensis sp. nov., Lactobacillus daoliensis sp. nov., Lactobacillus nangangensis sp. nov., Lactobacillus daowaiensis sp. nov., Lactobacillus dongliensis sp. nov., Lactobacillus songbeiensis sp. nov. and Lactobacillus kaifaensis sp. nov., isolated from traditional Chinese pickle.

Seven novel lactic acid bacterial strains, isolated from traditional Chinese pickle, were characterized using a polyphasic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, determination of DNA G+C content, determination of average nucleotide identity (ANI), in silico DNA-DNA hybridization (isDDH), fatty acid methyl ester (FAME) analysis and an analysis of phenotypic features. Strains 382-1T, 116-1AT, 381-7T, 203-3T, 218-3T and 398-2T were phylogenetically related to the type strains of Lactobacillus plantarumsubsp. plantarum, Lactobacillus plantarumsubsp. argentoratensis, Lactobacillus pentosus, Lactobacillus paraplantarum, Lactobacillus fabifermentans, Lactobacillus herbarum, Lactobacillus mudanjiangensis, Lactobacillus xiangfangensis, Lactobacillus plajomi and Lactobacillus modestisalitolerans, having 97.1-99.9 % 16S rRNA gene sequence similarities, less than 89.9 % pheS gene sequence similarities, less than 98.0 % rpoA gene sequence similarities, less than 91.2 % ANI values and less than 43.3 % isDDH values. Strain 778-3T was phylogenetically related to the type strains of Lactobacillus hokkaidonensis, Lactobacillus wasatchensis, Lactobacillus oligofermentans, Lactobacillus nenjiangensis, Lactobacillus vaccinostercus and Lactobacillus suebicus, exhibiting 97.0-99.4 % 16S rRNA gene sequence similarities, 78.2-82.1 % pheS gene sequence similarities, 80.0-91.5 % rpoA gene sequence similarities, less than 78.6 % ANI values and less than 22.9 % isDDH values. Based upon the data of polyphasic characterization obtained in the present study, seven novel species, Lactobacillus pingfangensis sp. nov., Lactobacillus daoliensis sp. nov., Lactobacillus nangangensis sp. nov., Lactobacillus daowaiensis sp. nov., Lactobacillus dongliensis sp. nov., Lactobacillus songbeiensis sp. nov. and Lactobacillus kaifaensis sp. nov., are proposed and the type strains are 382-1T (=NCIMB 15187T=CCM 8935T=LMG 31176T), 116-1AT (=NCIMB 15181T=CCM 8934T=LMG 31171T), 381-7T (=NCIMB 15186T=CCM 8930T), 203-3T (=NCIMB 15183T=CCM 8933T=LMG 31172T), 218-3T (=NCIMB 15184T=CCM 8932T=LMG 31173T), 398-2T (=NCIMB 15189T=CCM 8931T=LMG 31174T) and 778-3T (=NCIMB 15191T=CCM 8929T=LMG 31177T), respectively.

Enterococcus pingfangensis sp. nov., Enterococcus dongliensis sp. nov., Enterococcus hulanensis sp. nov., Enterococcus nangangensis sp. nov. and Enterococcus songbeiensis sp. nov., isolated from Chinese traditional pickle juice.

Five novel lactic acid bacterial strains, isolated from Chinese traditional pickle juice, were characterised by a polyphasic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, determination of DNA G+C content, average nucleotide identity (ANI) analysis, in silico DNA-DNA hybridisation (isDDH), fatty acid methyl ester (FAME) analysis and an analysis of phenotypic features. Strains 241-2-2T, 63-4T and 190-7T were closely related to the type strains of Enterococcus devriesei, Enterococcus viikkiensis, Enterococcus pseudoavium, Enterococcus xiangfangensis, Enterococcus avium, Enterococcus malodoratus, Enterococcus raffinosus and Enterococcus gilvus, having 99.1-99.9 % 16S rRNA gene sequence similarities. Strain 94-2T was distantly related to the type strains of Enterococcus phoeniculicola, Enterococcus rivorum and Enterococcus faecalis, having 95.2-96.1 % 16S rRNA gene sequence similarities. Strain 85-4T was distantly related to the type strains of Enterococcus casseliflavus, Enterococcus gallinarum, Enterococcus dispar, Enterococcus canintestini, Enterococcus saigonensis, Enterococcus diestrammenae, Enterococcus asini, Enterococcus cecorum and Enterococcus columbae, having 95.7-97.8 % 16S rRNA gene sequence similarities. Less than 91.5 % ANI and 45.3 % isDDH values between strains 241-2-2T, 63-4T, 190-7T, 94-2T, 85-4T and type strains of phylogenetically related species showed that they represent five new species within the genus Enterococcus. Based upon the data of polyphasic characterisation obtained in the present study, five novel species, Enterococcus pingfangensis sp. nov., Enterococcus dongliensis sp. nov., Enterococcus hulanensis sp. nov., Enterococcus nangangensis sp. nov. and Enterococcus songbeiensis sp. nov., are proposed and the type strains are 241-2-2T (=NCIMB 15185T=CCM 8921T=LMG 31181T), 63-4T (=NCIMB 15178T=CCM 8922T), 190-7T (=NCIMB 15200T=CCM 8949T), 94-2T (=NCIMB 15180T=CCM 8920T=LMG 31180T) and 85-4T (=NCIMB 15179T=CCM 8923T=LMG 31183T), respectively.