Sp Strain Research Articles

Phylogenetic analysis of Arthrospira strains from Ordos based on 16S rRNA

To study the growth rate of different Arthrospira strains, three species of Arthrospira from Ordos alkaline lake, labeled as sp.DD, sp.ER, sp.FB, one species of Arthrospira from Hasu Sea in Hohhot, labeled as sp.HS, another purified strain labeled as sp.QD donated by the Ocean University of China had been collected. The first four need to be further isolated and purified in culture. The growth curves of all strains were plotted. Subsequently, 16S rRNA sequences were amplified and sequenced in an attempt to study taxonomic relationships. The results showed that the growth rate was increased in the first 9 days, and sp.DD had the highest growth rate. Analysis of the sequencing results revealed that sp.HS had 99.79% homology with Arthrospira platensis strain Sp-2, sp.DD had 99.69% homology with Arthrospira platensis FACHB834, sp.QD had 99.54% homology with Arthrospira platensis F3S, sp.ER had 99.79% homology with Arthrospira erdosensis ‘Inner Mongolia’, sp.FB had 99.74% homology with Arthrospira erdosensis ‘Inner Mongolia’. Phylogenetic analysis indicated that sp.HS was closely related to Arthrospira platensis strain Sp-2; sp.DD and sp.QD had a close genetic relationship; sp.ER and sp.FB had a close genetic relationship. In conclusion, these findings provide a theoretical basis for the further development and reproduction of dominant algae species in Inner Mongolia through biological analysis of Arthrospira.

Comparative Analysis of Bacillariophyceae Chloroplast Genomes Uncovers Extensive Genome Rearrangements Associated with Speciation.

The Bacillariophyceae is a species-rich, ecologically significant class of Bacillariophyta. Despite their critical importance in marine ecosystems as primary producers and in the development of harmful algal blooms (HABs), taxonomic research on Bacillariophyceae species has been hindered because of their limited morphological features, plasticity of morphologies, and the low resolution of common molecular markers. Hence molecular markers with improved resolution are urgently needed. Organelle genomes, which can be constructed efficiently with the recent development of high throughput DNA sequencing technologies and the advancement of bioinformatics tools, have been proposed as super barcodes for their higher resolution for distinguishing different species and intra-species genomic variations. In this study, we tested the value of full-length chloroplast genomes (cpDNAs) as super barcodes for distinguishing diatom species, by constructing cpDNAs of 11 strains of the class Bacillariophyceae, including Nitzschia ovalis, Nitzschia traheaformis, Cylindrotheca spp., Psammodictyon constrictum, Bacillaria paxillifer, two strains of Haslea tsukamotoi, Haslea avium, Navicula arenaria, and Pleurosigma sp. Comparative analysis of cpDNAs revealed that cpDNAs were not only adequate for resolving different species, but also for enabling recognition of high levels of genome rearrangements between cpDNAs of different species, especially for species of the genera Nitzschia, Cylindrotheca, Navicula and Haslea. Additionally, comparative analysis suggested that the positioning of species in the genus Haslea should be transferred to the genus Navicula. Chloroplast genome-based evolutionary analysis suggested that the Bacillariophyceae species first appeared during the Cretaceous period and the diversity of species rose after the mass extinction about 65 Mya. This study highlighted the value of cpDNAs in research on the biodiversity and evolution of Bacillariophyceae species, and, with the construction of more cpDNAs representing additional genera, deeper insight into the biodiversity and evolutionary relationships of Bacillariophyceae species will be gained.

The attenuated Mycoplasma bovis strain promotes apoptosis of bovine macrophages by upregulation of CHOP expression.

Mycoplasma bovis (M. bovis) is one of the major pathogens in the bovine respiratory disease complex, which includes pneumonia, mastitis, and arthritis and causes a great economic loss in the cattle industry. In China, a live-attenuated vaccine strain M. bovis P150 was obtained by a continuous culture of the wild-type strain M. bovis HB0801 (P1) in vitro for 150 passages. Using the infected bovine macrophage cell line BoMac, this work attempted to investigate the mechanism of P150 attenuation and protective immune response. To begin, we show that M. bovis P150 effectively triggered cytotoxicity and apoptosis in BoMac, although with lower intracellular survival than P1. The transcriptomes of BoMac after infection with M. bovis strains P1 and P150 were sequenced, and bioinformatic analysis identified 233 differentially expressed genes (DEGs), with 185 upregulated and 48 downregulated. Further Gene Ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analyses revealed that the majority of the DEGs were linked to CHOP complex, MAP kinase phosphatase activity and were involved in the IL-17 signaling pathway in immune response, MAPK signaling pathway in signal transduction, and p53 signaling pathway in cell growth and death. Among them, the level of C/EBP homologous protein (CHOP) was significantly upregulated in P150-infected BoMac compared to P1-infected cells at different time points, along with its upstream and downstream genes phosphorylated-PERK, phosphorylated-EIF2α, ATF4, and GADD45A increased in the PERK-dependent ER stress response. The role of CHOP in apoptosis was further verified by M. bovis-induced siCHOP knockdown in BoMac cells. The results showed that CHOP knockdown enhanced P150-induced apoptosis and dramatically increased the M. bovis P1 and P150 intracellular survival, particularly for P150. These data suggest that P150 infection upregulates CHOP expression, which can increase apoptosis and mediate a crosstalk between ER stress and apoptosis during infection, and hence, contribute to high cytotoxicity and low intracellular survival.

Siderophore production in pseudomonas SP. strain SP3 enhances iron acquisition in apple rootstock.

The purpose of this study was to analyse the effects of siderophore-producing bacteria and bacterial siderophore on the iron nutrition of apple rootstocks under iron-deficient conditions. We isolated three Pseudomonas strains, SP1, SP2 and SP3 from the rhizosphere of the Fe-efficient apple rootstocks using the chrome azurol S agar plate assay. We found that all three strains had the ability to secrete indole acetic acid-like compounds and siderophores, especially SP3. When Fe-inefficient rootstocks treated with SP3 were grown in alkaline soil, an increase in the biomass, root development, and Fe concentration was observed in the plants. In addition, SP3 secreted pyoverdine, a siderophore that can chelate Fe3+ to enhance the bioavailability of Fe for plants. We purified the pyoverdine from the SP3 culture supernatant. Hydroponic experiments were conducted with a Fe-deficient solution supplemented with pyoverdine, resulting in a reduction in the chlorosis caused by Fe deficiency and marked improvement in Fe uptake. Under iron-deficient conditions, Pseudomonas sp. strain SP3 can effectively promote apple rootstock growth and improve plant iron nutrition by secreting siderophores that enhance Fe availability. This study showed that plant growth-promoting rhizobacteria from Fe-efficient plants have the potential to improve iron nutrition in Fe-inefficient plants, and Fe-siderophore chelates can be used as an effective source of iron for apple plants. Based on these findings, it may be possible to develop biological agents such as siderophore-producing bacteria for sustainable agricultural and horticultural production.

Screening of high lipid content and productivity of microalgae under photoautotrophic cultivation for biodiesel production

AbstractMicroalgae are globally accepted source of fatty acids (FA) due to their cell's potential to efficiently accumulate triacylglycerides which contain a significant amount of fatty acids for biodiesel production. This study aimed to screen nine (09) green microalgae strains in the laboratory for their high biomass and lipid productivity. Various microalgae strains were cultured at 30°C, with a light to dark ratio of 16:8 h (10–12 klux of light intensity) and aeration supplementation of 2% CO2 for longer period of time. The fatty acids of all microalgae species which showed growth potential under photoautotrophic cultivation had similar fatty acids found in palm oil. The selected strains of Ankistrodesmus sp. IFRPD 1061 and Chlorella sp. IFRPD 1092 were used due to their high lipid productivity content, were cultivated in 100 L NS III medium in an open raceway pond, aerated with 2% v/v CO2. The biomass concentration and productivity, lipid concentration and productivity of Ankistrodesmus sp. IFRPD 1061 (0.865 g/L, 76.250 mg/L/d, 0.167 g/L, and 14.685 mg/L/d, respectively) were found to be higher than those of Chlorella sp. IFRPD 1092 (0.433 g/L, 31.250 mg/L/d, 0.137 g/L, and 8.528 mg/L/d, respectively). Ankistrodesmus sp. IFRPD 1061 showed potential results of growing microalgae under photoautotrophic cultivation for biodiesel production.

Synergism of Bradyrhizobium and Azospirillum baldaniorum improves growth and symbiotic performance in lima bean under salinity by positive modulations in leaf nitrogen compounds

Salinity causes serious agricultural losses, and inoculation with plant growth-promoting bacteria (PGPB) represents an alternative to mitigate this and other abiotic stresses. This study aimed to investigate the effectiveness of five different rhizobia strains (B. elkanii BR 2003, B. pachyrhizi BR 3262, B. yuanmingense BR 3267, B. paxllaeri BR 10398, and B. icense BR 10399) and one Azospirillum baldaniorum strain (Sp245), alone or in combination, in inducing salt stress tolerance in lima bean. Plants were grown from seeds and inoculated with different rhizobia strains, A. baldaniorum, or both (coinoculation) and exposed to 0 (control) and 75 mM NaCl. Uninoculated plants were used as absolute control. Salinity negatively affects the absolute growth rate, root length, plant biomass (shoot, root, and nodules), number of nodules, and photosynthetic pigments. However, plants coinoculated with rhizobia and A. baldaniorum showed the highest value for these parameters. Salt stress provokes negative alterations in nitrogen compounds (nitrate, free ammonia, free amino acids, proline, and soluble protein). Nevertheless, these responses were minimized in plants coinoculated with rhizobia and A. baldaniorum — mainly BR 2003 and A. baldaniorum or BR 10399 and A. baldaniorum. The highest total N values were registered in plants coinoculated with BR 3267 and A. baldaniorum or BR 10399 and A. baldaniorum. Under salt stress, coinoculated plants show a minor increase in sodium and the highest potassium content values, nitrogen derived from the atmosphere, and nitrogen fixation efficiency than plants inoculated with rhizobia. Our findings indicate that lima bean coinoculation with rhizobia and A. baldaniorum was beneficial under salinity and improved salt tolerance. Synergistic interactions between rhizobia and A. baldaniorum, mainly among BR 2003 and A. baldaniorum and BR 10399 and A. baldaniorum, attenuated the deleterious effects of salt stress on lima bean. Furthermore, nitrogen fixation in root nodules was improved in coinoculated plants. Our results may be useful to develop adequate combinations of inoculants with rhizobia and A. baldaniorum for improving the growth and symbiotic performance of lima bean and others legumes under stressful conditions.

Effect of microbial-induced calcite precipitation towards strength and permeability of peat

Peat is known as problematic ground with low bearing capacity and extensively high compressibility. Bio-cementation or commonly known as microbial-induced calcite precipitation (MICP) has been recently introduced as a ground improvement alternative for peat under waterlogged condition. Using isolated bacteria strains P19 and P21 from tropical peat, it is found that unconfined compression strength (UCS) increases with bacteria concentration at a reducing rate. A maximum unconfined compressive strength of 82.05 kPa was measured with bacteria strain P21 at 108 CFU/mL. For the range of cementation reagent varying from 0.1 to 4.0 mol/kg, the largest strength improvement occurred at 1 mol/kg and 2 mol/kg using indigenous bacteria and bacteria strain P21, respectively, for peat with sand content of 25%. At 4.0 mol/kg, the cementation reagent has detrimental effect to MICP resulting in significant reduction in strength. Due to MICP, the UCS of peat increases with sand content. Calcium carbonate precipitation results in a reduction of permeability and an increment of strength of peat–sand mixture under a submerged condition up to 28 days.

Biochar amended microbial conversion of C1 gases to ethanol and butanol: Effects of biochar feedstock type and processing temperature

Biochar feedstock and production method affects its physicochemical properties and subsequent application. This study investigated the effects of biochar from switchgrass (SGB) and poultry litter (PLB) produced at 350 and 700 °C on alcohol formation using CO:CO2:H2 (40:30:30) by Clostridium carboxidivorans (P7) and C. ragsdalei (P11). Fermentations were performed in 250 mL bottles with a 50 mL working volume at 37 °C. Strains P7 and P11 produced 1.2- to 2.2-fold more alcohol and consumed 1.2- to 1.9-fold more syngas using biochars made at 700 °C compared to 350 °C. Both strains also produced 1.4- to1.9-fold more alcohol with both biochars made at 700 °C compared to control without biochar. Strain P11 produced 1.1- and 1.6-fold more alcohol and fatty acids, respectively, in medium with PLB made at 700 °C compared to strain P7. These results provide guidance towards the selection of biochar type and production temperature to improve syngas fermentation.

Crude oil hydrocarbon degradation efficiency of indigenous bacterial strains isolated from contaminated sites in Nigeria

The crude oil degradation potential of bacterial isolates from three contaminated sites in Nigeria were investigated. Seven bacterial isolates namely Pseudomonas aeruginosa strain W15, Pseudomonas aeruginosa strain N3R, Serratia marcescens strain N4, Providencia vermicola strain W8, Serratia marcescens strain W13, Pseudomonas aeruginosa strain W11 and Pseudomonas protegens strain P7 were isolated and identified using molecular methods. Isolates N4, N3 and W13 showed higher % total petroleum hydrocarbon (TPH) degradation of 79.26%, 78.96% and 78.69% respectively than W15, P7, W8 and W11 with % TPH degradation of 68.96%, 62.14%, 59.75% and 59.00% respectively. W13 showed the fastest degradation rate with 78.72% within the first 14 days of incubation; however, after the 14th day, there was no progressive change in % TPH. W11 showed degradation of wider range of hydrocarbon components originally in the crude oil as well as the complete degradation of most intermediates formed. The isolates showed good degradation potentials for bioremediation applications.

КОМБІНОВАНІ РОЗЧИНИ НЕРОЗЧИННИХ У ВОДІ БІОЛОГІЧНО АКТИВНИХ СПОЛУК З ВИКОРИСТАННЯМ БІОСУРФАКТАНТІВ

Solutions of biologically active substances insoluble in water (thiosulfoesters, anthraquinone derivatives, pyrazoline) were obtained using a combined solvent and biogenic surfactants. For this purpose were used rhamnolipids, their biocomplex with polysaccharides synthesized by the Pseudomonas SP strain. PS-17, trehalosolipids of the Rhodococcus erythropolis AU-1 strain, and the commercial biosurfactant surfactant. Aqueous dispersed systems of the studied substances were also obtained. The research results have prospects for use in pharmacy.

Whole-Genome Analysis Reveals That Bacteriophages Promote Environmental Adaptation of Staphylococcus aureus via Gene Exchange, Acquisition, and Loss.

The study of bacteriophages is experiencing a resurgence owing to their antibacterial efficacy, lack of side effects, and low production cost. Nonetheless, the interactions between Staphylococcus aureus bacteriophages and their hosts remain unexplored. In this study, whole-genome sequences of 188 S. aureus bacteriophages—20 Podoviridae, 56 Herelleviridae, and 112 Siphoviridae—were obtained from the National Center for Biotechnology Information (NCBI, USA) genome database. A phylogenetic tree was constructed to estimate their genetic relatedness using single-nucleotide polymorphism analysis. Comparative analysis was performed to investigate the structural diversity and ortholog groups in the subdividing clusters. Mosaic structures and gene content were compared in relation to phylogeny. Phylogenetic analysis revealed that the bacteriophages could be distinguished into three lineages (I–III), including nine subdividing clusters and seven singletons. The subdividing clusters shared similar mosaic structures and core ortholog clusters, including the genes involved in bacteriophage morphogenesis and DNA packaging. Notably, several functional modules of bacteriophages 187 and 2368A shared more than 95% nucleotide sequence identity with prophages in the S. aureus strain RJ1267 and the Staphylococcus pseudintermedius strain SP_11306_4, whereas other modules exhibited little nucleotide sequence similarity. Moreover, the cluster phages shared similar types of holins, lysins, and DNA packaging genes and harbored diverse genes associated with DNA replication and virulence. The data suggested that the genetic diversity of S. aureus bacteriophages was likely due to gene replacement, acquisition, and loss among staphylococcal phages, which may have crossed species barriers. Moreover, frequent module exchanges likely occurred exclusively among the subdividing cluster phages. We hypothesize that during evolution, the S. aureus phages enhanced their DNA replication in host cells and the adaptive environment of their host.

Double-edged effects of elevating temperature on the aging of exogenous arsenic in flooded paddy soils

Temperature variation can have a significant impact on arsenic (As) bioavailability in paddy soils. However, details regarding the transformation of exogenous As during the aging process in paddy soils at various temperatures remain unclear. This work investigated the effects of temperature on the As extractability and As species transformation of three paddy soils spiked with exogenous arsenate at 60 mg kg−1 under flooded aging and explored the related chemical and microbial mechanisms. The results showed that 0.05 M NH4H2PO4-extractable As decreased over time during flooded aging for 192 days, and it decreased by approximately one-third at 35 °C compared with 15 °C and 25 °C at the same aging time, indicating that higher temperatures facilitated the decrease in As extractability. As(V) reduction mainly occurred at 35 °C because the abundance and As(V)-reducing capacity of the predominant indigenous bacteria, the Bacillus sp strains, and the abundance of the arrA gene were significantly higher than those at 15 °C and 25 °C. The reduction of As(V) to As(III) and aging occurred simultaneously. The kinetic models were established, and the rate constants of the reduction and aging processes were obtained. Soil properties significantly affected the aging and reduction processes of extractable As(V). Our study indicated that elevating temperature had dual effects on the environmental risk of As in the flooded aging process. The previous definition of “aging” based on cationic metals needs to be updated according to the transformation characteristics of As species in flooded conditions. Our results addressed the necessity of impeding the reduction of As(V) in paddy soils under global warming.

Phosphate solubilizing epilithic and endolithic bacteria isolated from clastic sedimentary rocks, Murree lower Himalaya, Pakistan.

Rock microbes are capable to solubilize phosphate present in the rocks.. In this study, we focused on the isolation of phosphate solubilizing bacteria from rocks of Murree, Pakistan. Both endolithic and epilithic bacteria were screened for phosphate solubilization. Three bacterial strains were selected based on halozone formation inNational Botanical Research Institute for phosphate) medium supplemented with TCP (tribasic calcium phosphate). The solubilization index for these bacteria was recorded as 4.29, 4.03 and 3.99. The pH of the medium dropped from 7.0 to 4.0 after 5days with continuous shaking at 150rpm, which facilitate the phosphate solubilization. The strains P26, P4 and N27 were identified as Pseudomonas putida strain (KT004381), Pseudomonas grimontii (KT223621) and Alcaligenes faecalis (KT004385). Strain P26 showed maximum phosphate solubilization (367.54µg/ml), while P4 and N27 showed 321.88 and 291.36µg/ml after 3days of incubation. Such inorganic phosphate solubilization could be attributed to the organic acids production by bacteria. The presence of organic acids is determined by high-performance liquid chromatography. Three different types of acids, gluconic, oxalic and malic acid were the dominant acids found in the culture medium. It may be assumed that these bacteria can play a role in weathering of rocks as well. PSB is likely to serve as an efficient biofertilizer, especially in areas deficient in P to increase the overall performance of crops.

Effect of Holstein genotype on immune response to an intramammary Escherichia coli challenge

Selective breeding of US dairy cows since the mid-1960s has contributed to remarkable gains in milk yield per cow. This increased milk yield has been associated with an increase in health issues. Since 1964, the University of Minnesota has selectively bred a Holstein herd to maintain genetically static, unselected Holsteins (UH). Comparison of these UH cows with contemporary Holsteins (CH) has demonstrated that the UH cows not only produce less milk but also have fewer health concerns than their CH herdmates. The objective of this study was to determine the effects of Holstein genotype on innate immune response in an experimental intramammary Escherichia coli challenge model. Primiparous UH (n = 5) and CH (n = 7) cows received 430 cfu of E. coli strain P4 in 1 quarter. Blood and affected quarter milk samples were collected at 0, 0.25, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 7, 9, and 11 d relative to E. coli infusion. Rectal temperatures were recorded at each milking through d 4 of the experiment. Milk bacterial counts, somatic cell count and BSA concentrations, complete blood cell counts, rectal temperature, and serum and milk whey cytokine (IL-1β and IL-6) concentrations were used as metrics to determine infection severity. Longitudinal (repeated) data were analyzed using general linear models with PROC MIXED with day of study as the repeated effect. Whole blood transcriptomes were generated by RNA sequencing. Transcripts with a false discovery rate of P < 0.05 and a delta log2 expression value greater than 0.7 or less than -0.7 were used for functional enrichment analysis. Bacterial counts were consistently greater in milk from CH than UH cows from d 0.25 through d 2.5. Milk somatic cell count increased within 6 h (d 0.25) after E. coli administration in CH and UH cows but did not differ between genotypes after d 1. Rectal body temperature peaked at d 1 in CH and UH cows but was greater in CH cows. Milk BSA, IL-1β, and IL-6 concentrations were greater in CH than UH cows after E. coli administration. Blood lymphocyte and neutrophil counts were decreased at 0.5 and 1 d in CH but not in UH cows. The number of differentially expressed transcripts at each of the postinfusion sampling times was consistently greater (4- to 90-fold) in CH than in UH cows. A key difference between the immune reaction of the 2 genotypes was that the immune response to E. coli was largely contained within the mammary gland of the UH cows but became more systemic in the CH cows. These data demonstrate that UH cows exerted more effective control of E. coli infused into the mammary gland and thus support the hypothesis that selection practices since the mid-1960s have resulted in CH cows with an immune system that is less effective in fighting intramammary infections. Identification of genetic factors associated with enhanced immune functions that differ between the UH and CH cows could contribute to efforts to reintroduce or enhance beneficial components that have been lost or reduced in the CH cows since the mid-1960s.

In Vitro Application of Exogenous Fibrolytic Enzymes from Trichoderma Spp. to Improve Feed Utilization by Ruminants

Treating ruminant feeds with exogenous fibrolytic enzymes may potentially increase forage cell wall degradability and thus feed efficiency. In nature, fungi biosynthesize lignocellulolytic enzymes that can break down lignocellulosic material into its sugar components, thereby providing ready fermentable substrates. This work showed the in vitro fibrolytic activity of three Trichoderma strains (T. atroviride strain P1, T. afroharzianum strain T22, T. reesei strain T67). Total protein concentration and enzymatic (e.g., glucanase, cellulase, and xylanase) activities were determined in fungal culture filtrates after 7 and 14 days of growth on different fiber-based media. The enzymatic mixtures produced by Trichoderma spp. showed the highest concentration of fibrolytic enzymes and were added to industrial feed to test their ability to hydrolyze insoluble fibers. The supplementation of industrial feeds containing medium-fiber or low-fiber concentrates with T22 enzymes produced in the presence of lyophilized mushrooms and durum wheat fiber reduced hemicellulose concentration up to 33% and 24%, respectively. These results may offer novel opportunities to develop livestock feeds with improved fiber digestibility.

Comparative analysis of the Colistin resistance-regulating gene cluster in Klebsiella species

CrrAB two-component regulatory system is associated with colistin resistance in Klebsiella pneumoniae. Recently, some K. pneumoniae isolates lacking crrAB genes have been identified. In this study, we investigated the distribution and structural variation of the crrBAC-kexD cluster. To evaluate the structural variation of the crrBAC-kexD cluster, we explored 59 clinical K. pneumoniae isolates from Korea, and 508 whole genomes of K. pneumoniae and other strains of Klebsiella sp. Significant structural variations in crrBAC-kexD and its surrounding regions were identified among K. pneumoniae genomes. Within the genus Klebsiella, the cluster was identified only in K. pneumoniae, K. variicola, and K. quasipneumoniae, which form the K. pneumoniae complex. Among the 304 available K. pneumoniae genomes, an intact crrBAC-kexD cluster was identified in 178 isolates (58.6%), while the cluster was absent in 90 isolates (29.6%). Partial deletions within the cluster were identified in 22 genomes (7.2%). The most diverse structural patterns of the crrBAC-kexD cluster were observed in ST11 strains. Some clades lacked the crrBAC-kexD cluster. The crrBAC-kexD cluster was identified in the genomes of other bacterial species, including Citrobacter freundii and Enterobacter ludwigii. The crrBAC-kexD cluster is proposed to have been acquired by the ancestor of the K. pneumoniae complex from other bacterial species and the cluster may have been lost and re-acquired repeatedly in K. pneumoniae strains according to the phylogenetic analysis. The dynamic evolution of the crrBAC-kexD cluster suggests that it may have other roles, in addition to colistin resistance, in bacterial physiology.

Role of Lactobacillus plantarum with antioxidation properties on Chinese sausages

Excessive protein and lipid oxidation can lead to the deterioration of sausage quality, especially flavour. Functional starter culture increases their attractiveness through sensory qualities and a potential healthpromoting effect. It was investigated that Lactobacillus plantarum (L.plantarum) strains with antioxidation properties effected on the flavour and oxidation of fermented sausage in the study. The group treated with natural fermentation was used to as control (control group). Results showed that P3 and P3-M2 had the ability of reducing lipid and protein oxidation during fermentation. The increase of lipoxygenase activity and thiobarbituric acid reactive substances (TBARs) values were retarded. Sulfhydryl group contents were significantly higher than those in the control. Furthermore, changes in protein bands were confirmed with the less protein oxidation with P3-M2 than P3 and the control. Accordingly, strain P3 and P3-M2 significantly enhanced the types and relative contents of esters after fermented, indicating that strain P3 and P3-M2 contributed to the production of flavour substances. These results revealed that L.plantarum strains with antioxidation properties were a promising approach in inhibiting lipid and protein oxidation of Chinese sausage, maintaining the stable natural structure of protein, simultaneously improve the quality of sausage and promote the sausage to form a better flavour.

Comparative genome analysis of Pasteurella multocida serogroup B:2 strains causing haemorrhagic septicaemia (HS) in bovines

Pasteurella multocida, a Gram-negative bacterium with ubiquitous nature, is known to affect wide range of host species worldwide with varied clinical manifestations including haemorrhagic septicaemia (HS) in bovines. Although, HS causing P. multocida strains were identified and characterized by conventional tools and PCR assays, diverse strains are indistinguishable by these tools in the face of disease outbreaks. In this study, draft genomes of three virulent P. multocida serotype B:2 strains (NIVEDIPm32, NIVEDIPm34 and NIVEDIPm35) were analyzed following whole genome sequencing, assembly, annotation and compared them with existing global genomes (n = 43) of bovine origin in the database. Three draft genomes of NIVEDIPm strains consisted of 40–52 contigs with GC content of ∼40.4%. The genome size and predicted genes content was ∼2.3 Mb and 2181–2189, respectively. Besides, the presence of various mobile genetic elements, antimicrobial resistance genes and biofilm related genes suggested their vital roles in virulence; further, adaptation to the host immune system as well as host pathogen interaction. Multi locus sequence analysis based on RIRDC scheme showed the presence of ST122 in all the three strains. wgMLST based phylogenic analysis suggested that HS causing Indian virulent field strains differed geographically and showed diversity from existing HS vaccine strain P52. The phylogenetic tree revealed that North Indian strains share high similarity with strains of Pakistan than South Indian Strain. Notably, a high divergence of SNPs between the HS causing circulating virulent strains of India and current HS vaccine strain P52 suggested an imminent need for relook in to HS vaccination strategy for livestock in India.

Structure and biosynthesis of carotenoids produced by a novel Planococcus sp. isolated from South Africa

BackgroundThe genus Planococcus is comprised of halophilic bacteria generally reported for the production of carotenoid pigments and biosurfactants. In previous work, we showed that the culturing of the orange-pigmented Planococcus sp. CP5-4 isolate increased the evaporation rate of industrial wastewater brine effluent, which we attributed to the orange pigment. This demonstrated the potential application of this bacterium for industrial brine effluent management in evaporation ponds for inland desalination plants. Here we identified a C30-carotenoid biosynthetic gene cluster responsible for pigment biosynthesis in Planococcus sp. CP5-4 through isolation of mutants and genome sequencing. We further compare the core genes of the carotenoid biosynthetic gene clusters identified from different Planococcus species’ genomes which grouped into gene cluster families containing BGCs linked to different carotenoid product chemotypes. Lastly, LC–MS analysis of saponified and unsaponified pigment extracts obtained from cultures of Planococcus sp. CP5-4, revealed the structure of the main (predominant) glucosylated C30-carotenoid fatty acid ester produced by Planococcus sp. CP5-4.ResultsGenome sequence comparisons of isolated mutant strains of Planococcus sp. CP5-4 showed deletions of 146 Kb and 3 Kb for the non-pigmented and “yellow” mutants respectively. Eight candidate genes, likely responsible for C30-carotenoid biosynthesis, were identified on the wild-type genome region corresponding to the deleted segment in the non-pigmented mutant. Six of the eight candidate genes formed a biosynthetic gene cluster. A truncation of crtP was responsible for the “yellow” mutant phenotype. Genome annotation revealed that the genes encoded 4,4′-diapolycopene oxygenase (CrtNb), 4,4′- diapolycopen-4-al dehydrogenase (CrtNc), 4,4′-diapophytoene desaturase (CrtN), 4,4′- diaponeurosporene oxygenase (CrtP), glycerol acyltransferase (Agpat), family 2 glucosyl transferase 2 (Gtf2), phytoene/squalene synthase (CrtM), and cytochrome P450 hydroxylase enzymes. Carotenoid analysis showed that a glucosylated C30-carotenoid fatty acid ester, methyl 5-(6-C17:3)-glucosyl-5, 6′-dihydro-apo-4, 4′-lycopenoate was the main carotenoid compound produced by Planococcus sp. CP5-4.ConclusionWe identified and characterized the carotenoid biosynthetic gene cluster and the C30-carotenoid compound produced by Planococcus sp. CP5-4. Mass-spectrometry guided analysis of the saponified and unsaponified pigment extracts showed that methyl 5-glucosyl-5, 6-dihydro-apo-4, 4′-lycopenoate esterified to heptadecatrienoic acid (C17:3). Furthermore, through phylogenetic analysis of the core carotenoid BGCs of Planococcus species we show that various C30-carotenoid product chemotypes, apart from methyl 5-glucosyl-5, 6-dihydro-apo-4, 4′-lycopenoate and 5-glucosyl-4, 4-diaponeurosporen-4′-ol-4-oic acid, may be produced that could offer opportunities for a variety of applications.

Selective removal of the non-herbicidal (S)-enantiomer of dichlorprop from agricultural soil by an in-situ enriched consortium

Rac-dichlorprop [(R,S)-2-(2,4-dichlorophenoxy)propanoic acid], a commonly used chiral phenoxyalkanoic acid herbicide, is frequently detected in various environments, posing threats to human health and environmental safety. The (S)-enantiomer of dichlorprop shows a lower herbicidal activity but a higher toxicity as compared to the (R)-enantiomer of dichlorprop. It is of great importance to enantioselectively remove the non-herbicidal (S)-dichlorprop from the rac-dichlorprop in the application sites. In this study, the dissipation of (R)- and (S)-dichlorprop in an agricultural soil was investigated. We found that the dissipation rate of (S)-dichlorprop was higher than that of (R)-dichlorprop in an agricultural soil. Although the influences of different enantiomers of dichlorprop on soil bacterial community were not different, Sphingobium was significantly enriched in the (S)-dichlorprop-treated soil as compared to (R)-dichlorprop, indicating that Sphingobium species played important roles in the dissipation of (S)-dichlorprop. Furthermore, a bacterial consortium containing Sphingobium sp. strain SP-2 (enantioselectively transforming (S)-dichlorprop to 2,4-dichlorophenol) and Sphingopyxis sp. strain DP-21 (mineralizing 2,4-dichlorophenol), which synergistically mineralized the (S)-enantiomer of dichlorprop, was isolated from the agricultural soil. Bioaugmentation with the in-situ isolated consortium (strains SP-2 and DP-21) resulted in the enhanced removal of the non-herbicidal (S)-dichlorprop from the soil and in the adaptation of the inoculated degrading strains. This study offers new insights into the enhanced bioremediation of the non-herbicidal (S)-dichlorprop contaminated soils while simultaneously ensuring environmental safety and crop yield.