Characterization and shelf stability of sweetened condensed milk formulated with different sucrose substitutes during storage.

Sweetened condensed milk (SCM) is a value-added milk product with extended shelf life and high levels of nutrition. The high level of sucrose may lead to health problems. Many studies have focused on the reduction of sucrose but seldomly on different combination of sugar substitutes. This study aims to find an ideal sucrose substitution through physiochemical, microbiological and sensory properties of SCM under different storage times. The results demonstrated that substitution with 20% trehalose, 5% lactulose and 15% erythritol resulted in similar sensory and color as control group. The volatile flavor analysis showed that substitution with 30% trehalose, 5% lactulose and 5% polyols was the most similar and hexanoic acid was the symbolistic flavor. Sucrose replacement increased the antibacterial effect and Staphylococcus, Penicillium, Apiotrichum and Candida were widely present. Substitution with 30% trehalose, 5% lactulose and 5% polyols resulted in the most similar water activity, texture, aroma and microbial diversity.

D-glucose overflow metabolism in an evolutionary engineered high-performance D-xylose consuming Saccharomyces cerevisiae strain.

Co-consumption of D-xylose and D-glucose by Saccharomyces cerevisiae is essential for cost-efficient cellulosic bioethanol production. There is a need for improved sugar conversion rates to minimize fermentation times. Previously, we have employed evolutionary engineering to enhance D-xylose transport and metabolism in the presence of D-glucose in a xylose-fermenting S. cerevisiae strain devoid of hexokinases. Re-introduction of Hxk2 in the high performance xylose-consuming strains restored D-glucose utilization during D-xylose/D-glucose co-metabolism, but at rates lower than the non-evolved strain. In the absence of D-xylose, D-glucose consumption was similar to the parental strain. The evolved strains accumulated trehalose-6-phosphate during sugar co-metabolism, and showed an increased expression of trehalose pathway genes. Upon the deletion of TSL1, trehalose-6-phosphate levels were decreased and D-glucose consumption and growth on mixed sugars was improved. The data suggest that D-glucose/D-xylose co-consumption in high-performance D-xylose consuming strains causes the glycolytic flux to saturate. Excess D-glucose is phosphorylated enters the trehalose pathway resulting in glucose recycling and energy dissipation, accumulation of trehalose-6-phosphate which inhibits the hexokinase activity, and release of trehalose into the medium.

Trehalose as quantitative biomarker for in vivo diagnosis and treatment follow-up in cryptococcomas.

Brain lesions caused by Cryptococcus neoformans or C. gattii (cryptococcomas) are typically difficult to diagnose correctly and treat effectively, but rapid differential diagnosis and treatment initiation are crucial for good outcomes. In previous studies, cultured cryptococcal isolates and ex vivo lesion material contained high concentrations of the virulence factor and fungal metabolite trehalose. Here, we studied the in vivo metabolic profile of cryptococcomas in the brain using magnetic resonance spectroscopy (MRS) and assessed the relationship between trehalose concentration, fungal burden, and treatment response in order to validate its suitability as marker for early and noninvasive diagnosis and its potential to monitor treatment in vivo. We investigated the metabolites present in early and late stage cryptococcomas using in vivo 1H MRS in a murine model and evaluated changes in trehalose concentrations induced by disease progression and antifungal treatment. Animal data were compared to 1H and 13C MR spectra of Cryptococcus cultures and in vivo data from 2 patients with cryptococcomas in the brain. In vivo MRS allowed the noninvasive detection of high concentrations of trehalose in cryptococcomas and showed a comparable metabolic profile of cryptococcomas in the murine model and human cases. Trehalose concentrations correlated strongly with the fungal burden. Treatment studies in cultures and animal models showed that trehalose concentrations decrease following exposure to effective antifungal therapy. Although further cases need to be studied for clinical validation, this translational study indicates that the noninvasive MRS-based detection of trehalose is a promising marker for diagnosis and therapeutic follow-up of cryptococcomas.

Compositional differences in hybrids between protoporphyrinogen IX oxidase (PPO)-inhibiting herbicide-resistant transgenic rice and weedy rice accessions.

We characterized the metabolites in grains of transgenic protoporphyrinogen IX oxidase-inhibiting herbicide-resistant rice and weedy accessions using GC-MS and examined whether the chemical composition of their hybrids differed from that of the parents. We found that the metabolite profiles of transgenic rice and weedy rice were clearly separated. Although the metabolite profiles of F2 progeny were partially separated from their parents, zygosity did not affect the profiles. The F2 progeny had similar or intermediate levels of most major nutritional components compared with their parents. However, levels of galactopyranose, trehalose, xylofuranose, mannitol, and benzoic acid were higher in the F2 progeny. Some fatty acids and organic acids also showed prominent quantitative differences between the F2 progeny and the parents. Changes in the metabolite levels of transgenic crop-weed hybrids compared to their parents might influence not only the ecological consequences of the hybrids, but also the nutritional quality and food safety.

The secreted acid trehalase encoded by the CgATH1 gene is involved in Candida glabrata virulence.

BACKGROUND: Candida glabrata yeast is the second cause of candidiasis worldwide. Differs from other yeasts since assimilates only glucose and trehalose (a characteristic used in rapid identification tests for this pathogen) by secreting into the medium a highly active acid trehalase encoded by the CgATH1 gene. OBJECTIVE: This study aimed to characterise the function of the acid trehalase in the physiopathology of C. glabrata. METHODS: Gene deletion was performed to obtain a mutant ath1Δ strain, and the ability of the ath1Δ strain to grow in trehalase, or the presence of trehalase activity in the ath1Δ yeast cells, was verified. We also tested the virulence of the ath1Δ strain in a murine model of infection. FINDINGS: The ath1Δ mutant strain grows normally in the presence of glucose, but loses its ability to grow in trehalose. Due to the high acid trehalase activity present in wild-type cells, the cytoplasmic neutral trehalase activity is only detected in the ath1Δ strain. We also observed a significantly lower virulence of the ath1Δ strain in a murine model of infection with either normal or immunocompromised mice. MAIN CONCLUSIONS: The acid trehalase is involved in the hydrolysis of external trehalose by C. glabrata, and the enzyme also plays a major virulence role during infectivity.

Impact of maturation and growth temperature on cell-size distribution, heat-resistance, compatible solute composition and transcription profiles of Penicillium roqueforti conidia.

Penicillium roqueforti is a major cause of fungal food spoilage. Its conidia are the main dispersal structures of this fungus and therefore the main cause of food contamination. These stress resistant asexual spores can be killed by preservation methods such as heat treatment. Here, the effects of cultivation time and temperature on thermal resistance of P. roqueforti conidia were studied. To this end, cultures were grown for 3, 5, 7 and 10 days at 25 °C or for 7 days at 15, 25 and 30 °C. Conidia of 3- and 10-day-old cultures that had been grown at 25 °C had D56-values of 1.99 ± 0.15 min and 5.31 ± 1.04 min, respectively. The effect of cultivation temperature was most pronounced between P. roqueforti conidia cultured for 7 days at 15 °C and 30 °C, where D56-values of 1.12 ± 0.05 min and 4.19 ± 0.11 min were found, respectively. Notably, D56-values were not higher when increasing both cultivation time and temperature by growing for 10 days at 30 °C. A correlation was found between heat resistance of conidia and levels of trehalose and arabitol, while this was not found for glycerol, mannitol and erythritol. RNA-sequencing showed that the expression profiles of conidia of 3- to 10-day-old cultures that had been grown at 25 °C were distinct from conidia that had been formed at 15 °C and 30 °C for 7 days. Only 33 genes were upregulated at both prolonged incubation time and increased growth temperature. Their encoded proteins as well as trehalose and arabitol may form the core of heat resistance of P. roqueforti conidia.

Glucose transporter GLUT1 influences Plasmodium berghei infection in Anopheles stephensi.

BACKGROUND: Sugar-feeding provides energy for mosquitoes. Facilitated glucose transporters (GLUTs) are responsible for the uptake of glucose in animals. However, knowledge of GLUTs function in Anopheles spp. is limited. METHODS: Phylogenetic analysis of GLUTs in Anopheles stephensi was performed by the maximum likelihood and Bayesian inference methods. The spatial and temporal expression patterns of four Asteglut genes were analyzed by qPCR. The function of Asteglut1 was examined using a dsRNA-mediated RNA interference method. Transcriptome analysis was used to investigate the global influence of Asteglut1 on mosquito physiology. RESULTS: We identified 4 glut genes, Asteglut1, Asteglutx, Asteglut3 and Asteglut4 in An. stephensi. Asteglut1, Asteglut3 and Asteglut4 were mainly expressed in the midgut. Plasmodium berghei infection differentially regulated the expression of Asteglut genes with significant downregulation of Asteglut1 and Asteglut4, while upregulation of Asteglutx. Only knocking-down Asteglut1 facilitated Plasmodium berghei infection in An. stephensi. This might be due to the accumulation of glucose prior to blood-feeding in dsAsteglut1-treated mosquitoes. Our transcriptome analysis revealed that knockdown of Asteglut1 differentially regulated expression of genes associated with multiple functional clusters, especially those related to detoxification and immunity. The dysregulation of multiple pathways might contribute to the increased P. berghei infection. CONCLUSIONS: Our study shows that Asteglut1 participates in defense against P. berghei in An. stephensi. The regulation of Asteglut1 on vector competence might through modulating multiple biological processes, such as detoxification and immunity.

Content of xylose, trehalose and l-citrulline in cucumber fermentations and utilization of such compounds by certain lactic acid bacteria.

This research determined the concentration of trehalose, xylose and l-citrulline in fresh and fermented cucumbers and their utilization by Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus brevis and Lactobacillus buchneri. Targeted compounds were measured by HPLC and the ability of the lactobacilli to utilize them was scrutinized in fermented cucumber juice. Fresh cucumber juice was supplemented with trehalose, xylose and l-citrulline to observed mixed culture fermentations. Changes in the biochemistry, pH and colony counts during fermentations were monitored. Trehalose, xylose and l-citrulline were detected in fermentations to15.51 ± 1.68 mM, a fresh cucumber sample at 36.05 mM and in fresh and fermented cucumber samples at 1.05 ± 0.63 mM, respectively. Most of the LAB tested utilized trehalose and xylose in FCJM at pH 4.7. l-citrulline was utilized by L. buchneri and produced by other LAB. l-citrulline (12.43 ± 2.3 mM) was converted to ammonia (14.54 ± 3.60 mM) and the biogenic amine ornithine (14.19 ± 1.07 mM) by L. buchneri at pH 4.7 in the presence of 0.5 ± 0.2 mM glucose enhancing growth by 0.5 log CFU/mL. The use of a mixed starter culture containing L. buchneri aided in the removal of l-citrulline and enhanced the fermentation stability. The utilization of l-citrulline by L. buchneri may be a cause of concern for the stability of cucumber fermentations at pH 3.7 or above. This study identifies the use of a tripartite starter culture as an enhancer of microbial stability for fermented cucumbers.

Effect of freezing on the metabolic status of L3 larvae of Anisakis simplex s. s.

The fish-borne parasite, Anisakis simplex s. s., triggers a disease called anisakiasis, that is associated with a gastrointestinal infection. The Anisakis is also associated with allergic response which may lead to anaphylactic shock. The A. simplex s. s. L3 larvae may be freeze tolerant despite when the nematodes will be cooled rapidly to -20 °C according to the sanitary authorities of the USA and the EU. The aim of this work was to study the metabolic status of A. simplex s. s. L3 larvae when frozen in terms of viability, expression of genes involved in the nematodes' survival of freezing, as well content of carbohydrates which play a cryoprotective role in thermal stress and are the main source of energy. The levels of trehalose were significantly higher after slow freezing treatment (p < .0001), than the fast freezing (p < .002). The lower temperatures induce changes, especially in trehalose synthesis gene expression, genes responsible for oxidative metabolism, and chaperone proteins, but we cannot state clearly whether these changes occur during freezing, or because they are already prevalent during cold acclimation. The induction of mentioned genes seems to be a common trait of both cold- and dehydration tolerance.

The secreted acid trehalase encoded by the CgATH1 gene is involved in Candida glabrata virulence

BACKGROUND Candida glabrata yeast is the second cause of candidiasis worldwide. Differs from other yeasts since assimilates only glucose and trehalose (a characteristic used in rapid identification tests for this pathogen) by secreting into the medium a highly active acid trehalase encoded by the CgATH1 gene. OBJECTIVE This study aimed to characterise the function of the acid trehalase in the physiopathology of C. glabrata. METHODS Gene deletion was performed to obtain a mutant ath1Δ strain, and the ability of the ath1Δ strain to grow in trehalase, or the presence of trehalase activity in the ath1Δ yeast cells, was verified. We also tested the virulence of the ath1Δ strain in a murine model of infection. FINDINGS The ath1Δ mutant strain grows normally in the presence of glucose, but loses its ability to grow in trehalose. Due to the high acid trehalase activity present in wild-type cells, the cytoplasmic neutral trehalase activity is only detected in the ath1Δ strain. We also observed a significantly lower virulence of the ath1Δ strain in a murine model of infection with either normal or immunocompromised mice. MAIN CONCLUSIONS The acid trehalase is involved in the hydrolysis of external trehalose by C. glabrata, and the enzyme also plays a major virulence role during infectivity.

Chemical Fingerprint of 'Oblacinska' Sour Cherry (Prunuscerasus L.) Pollen.

The aim of this research was to analyze sugars and phenolics of pollen obtained from 15 different 'Oblacinska' sour cherry clones and to assess the chemical fingerprint of this cultivar. Carbohydrate analysis was done using high-performance anion-exchange chromatography (HPAEC) with pulsed amperometric detection (PAD), while polyphenols were analyzed by ultra-high-performance liquid chromatography-diode array detector-tandem mass spectrometry (UHPLC-DAD MS/MS) system. Glucose was the most abundant sugar, followed by fructose and sucrose. Some samples had high level of stress sugars, especially trehalose. Rutin was predominantly polyphenol in a quantity up to 181.12 mg/kg (clone III/9), with chlorogenic acid (up to 59.93 mg/kg in clone III/9) and p-coumaric acid (up to 53.99 mg/kg in clone VIII/1) coming after. According to the principal component analysis (PCA), fructose, maltose, maltotriose, sorbitol, and trehalose were the most important sugars in separating pollen samples. PCA showed splitting off clones VIII/1, IV/8, III/9, and V/P according to the quantity of phenolics and dissimilar profiles. Large differences in chemical composition of studied 'Oblacinska sour cherry' clone pollen were shown, proving that it is not a cultivar, but population. Finally, due to the highest level of phenolics, clones IV/8, XV/3, and VIII/1 could be singled out as a promising one for producing functional food and/or in medicinal treatments.

Characterization of trehalose lipids produced by a unique environmental isolate bacterium Rhodococcus qingshengii strain FF.

AIMS: The aim of this study was to elucidate the chemical properties and applications of trehalose lipids produced by Rhodococcus qingshengii strain FF and optimize its production yield. METHODS AND RESULTS: Strain FF was identified as R. qingshengii. It was observed to produce biosurfactants in the presence of n-hexadecane. The biosurfactants were identified as the mixture of trehalose triesters and trehalose tetraesters, mainly consisting of TrehC12 C3 C6 C12 :10, TrehC11 C8 C6 :6, TrehC11 C6 C4 :5 and TrehC6 C4 C6 :5 based on the analysis of thin layer chromatography, Fourier transform infrared and flight tandem mass spectrometry. The best carbon source and nitrogen source for producing trehalose lipids was the mixture of n-hexadecane and oleic acid (m : m = 1 : 1) and the organic nitrogen, urea. Under this condition, the production of trehalose lipids could reach 7·97 g l-1 . The crude trehalose lipids showed extremely high surface-active properties and were proven to promote the degradation of naphthalene. CONCLUSIONS: The trehalose lipids produced by R. qingshengii strain FF exhibited high surfactant activity under various conditions and were proven to promote the degradation of naphthalene. SIGNIFICANCE AND IMPACT OF THE STUDY: Rhodococcus qingshengii strain FF is a potential candidate for bioremediation. The trehalose lipids might be used as unique biosurfactants in cosmetic industries, biological formulations and other applications.

Enhanced Ion Yields Using High Energy Water Cluster Beams for Secondary Ion Mass Spectrometry Analysis and Imaging.

Previous studies have shown that the use of a 20 keV water cluster beam as a primary beam for the analysis of organic and bio-organic systems resulted in a 10-100 times increase in positive molecular ion yield for a range of typical analytes compared to C60 and argon cluster beams. This resulted in increased sensitivity to important lipid molecules in the bioimaging of rat brain. Building on these studies, the present work compares 40 and 70 keV water cluster beams with cluster beams composed of pure argon, argon and 10%CO2, and pure CO2. First, as previously, we show that for E/nucleon about 0.3 eV/nucleon water and nonwater containing cluster beams generate very similar ion yields, but below this value, the water beams yields of BOTH negative and positive "molecular" ions increase, in many cases reaching a maximum in the <0.2 region, with yield increases of ∼10-100. Ion fragment yields in general decrease quite dramatically in this region. Second, for water cluster beams at a constant E/nucleon, "molecular" ion yield increases with beam energy and hence cluster size due to increased sputter yield (ionization probability is constant). Third, as a consequence of the increased ion yield and the improved focusability using high-energy cluster beams, imaging in the 1 µm spatial resolution region is demonstrated on HeLa cells and rat brain tissue, monitoring molecules that were previously difficult to detect with other primary beams. Finally, the suggestion that the secondary ion emission zone has quasi-aqueous character seems to be sustained.

Ethanol Stimulates Trehalose Production through a SpoT-DksA-AlgU-Dependent Pathway in Pseudomonas aeruginosa.

Pseudomonas aeruginosa frequently resides among ethanol-producing microbes, making its response to the microbially produced concentrations of ethanol relevant to understanding its biology. Our transcriptome analysis found that genes involved in trehalose metabolism were induced by low concentrations of ethanol, and biochemical assays showed that levels of intracellular trehalose increased significantly upon growth with ethanol. The increase in trehalose was dependent on the TreYZ pathway but not other trehalose-metabolic enzymes (TreS or TreA). The sigma factor AlgU (AlgT), a homolog of RpoE in other species, was required for increased expression of the treZ gene and trehalose levels, but induction was not controlled by the well-characterized proteolysis of its anti-sigma factor, MucA. Growth with ethanol led to increased SpoT-dependent (p)ppGpp accumulation, which stimulates AlgU-dependent transcription of treZ and other AlgU-regulated genes through DksA, a (p)ppGpp and RNA polymerase binding protein. Ethanol stimulation of trehalose also required acylhomoserine lactone (AHL)-mediated quorum sensing (QS), as induction was not observed in a ΔlasR ΔrhlR strain. A network analysis using a model, eADAGE, built from publicly available P. aeruginosa transcriptome data sets (J. Tan, G. Doing, K. A. Lewis, C. E. Price, et al., Cell Syst 5:63-71, 2017, https://doi.org/10.1016/j.cels.2017.06.003) provided strong support for our model in which treZ and coregulated genes are controlled by both AlgU- and AHL-mediated QS. Consistent with (p)ppGpp- and AHL-mediated quorum-sensing regulation, ethanol, even when added at the time of culture inoculation, stimulated treZ transcript levels and trehalose production in cells from post-exponential-phase cultures but not in cells from exponential-phase cultures. These data highlight the integration of growth and cell density cues in the P. aeruginosa transcriptional response to ethanol.IMPORTANCEPseudomonas aeruginosa is often found with bacteria and fungi that produce fermentation products, including ethanol. At concentrations similar to those produced by environmental microbes, we found that ethanol stimulated expression of trehalose-biosynthetic genes and cellular levels of trehalose, a disaccharide that protects against environmental stresses. The induction of trehalose by ethanol required the alternative sigma factor AlgU through DksA- and SpoT-dependent (p)ppGpp. Trehalose accumulation also required AHL quorum sensing and occurred only in post-exponential-phase cultures. This work highlights how cells integrate cell density and growth cues in their responses to products made by other microbes and reveals a new role for (p)ppGpp in the regulation of AlgU activity.

Combinatorial impact of osmotic and heat shocks on the composition of membrane lipids and osmolytes in Aspergillus niger.

The combinatorial action of osmotic (OS) and heat (HS) shocks on the composition of soluble cytosol carbohydrates and membrane lipids was studied. For the first time it was demonstrated that the combinatorial effect of these shocks led to the non-additive response - an increase in the trehalose level, characteristic for HS, but at the same time suppression of glycerol production, uncharacteristic of the OS response. In addition, combinatorial action resulted in a new effect - increase in the mannitol level, which was not typical for the individual HS or OS responses. On the contrary, a general pattern of change was observed in the composition of membrane lipids in response to both individual HS and OS, and their combinations, which was a twofold increase in the proportion of phosphatidic acids. At the same time, the mechanism of alteration in the degree of unsaturation of membrane phospholipids was not involved in adaptation. The response to combinatorial shocks includes the accumulation of trehalose and mannitol, and increase in the proportion of phosphatidic acids in membrane lipids.

Sensitive analysis of trehalose-6-phosphate and related sugar phosphates in plant tissues by chemical derivatization combined with hydrophilic interaction liquid chromatography-tandem mass spectrometry.

Trehalose-6-phosphate (T6P) is an important signaling metabolite that is involved in many physiological processes. However, the mechanism of the biological functions of T6P is not fully understood. Quantification of T6P in plants will be beneficial to elucidate the mechanism. However, it is still a challenge to chromatographically separate and sensitively detect T6P and related sugar phosphates. In the current study, we developed a method for effective separation and sensitive detection of glucose-1-phosphate (G1P), glucose-6-phosphate (G6P), sucrose-6-phosphate (S6P) and T6P in plant tissues by chemical derivatization combined with hydrophilic interaction liquid chromatography-tandem mass spectrometry (ChD-HILIC-MS/MS). With this method, two pairs of isomers (G1P/G6P and S6P/T6P) could be well separated on a HILIC column and sensitively detected by MS with limits of detection (LODs) ranging from 0.1 to 0.6 ng mL-1. The developed method was successfully applied to the detection of endogenous G1P, G6P, S6P and T6P in small amounts of plant tissues, such as 1 mg fresh weight of Oryza sativa shoot.

Dynamic Analysis of Nucleosides and Carbohydrates during Developmental Stages of Cordyceps militaris in Silkworm (Bombyxmori).

Background: Cultured Cordyceps militaris is very popular. Objective: To gain dynamic insight into activity markers in fruiting body of Cordyceps militaris (C. militaris) in Bombyxmori (B. mori), also named silkworm. Methods: The development stages of samples at 3, 9, 12, 19, 27, and 33 days after inoculation (DAI) were collected. HPLC coupled with diode array detection and evaporative light-scattering detection method (HPLC-DAD-ELSD) was used to determine eight makers, including six nucleosides and two carbohydrates from the samples. Results: C. militaris cultured 33 DAI with fifth star silkworm larva could accumulate higher levels of cordycepin (13.43 mg/g) than the highest reported cordycepin (8.57 g/L). The contents of cordycepin, adenosine, and trehalose were gradually increased with the formation of C. militaris fruiting bodies on silkworm larva, while mannitol was decreased. The change of guanosine was similar to uracil. Conclusions: Results suggested that mannitol could be accumulated in a short period during mycelium growth and could metabolize and transform into energy store and trehalose during fruit body formation. The inosine in the insect was completely utilized and transformed. The synergistic formation of cordycepin and adenosine or differences in metabolized pathways are a great possibility according to the same trend. Highlights: This research offered some reference to further find a certain regularity or metabolic mechanism.

Chemoenzymatic radiosynthesis of 2-deoxy-2-[18F]fluoro-d-trehalose ([18F]-2-FDTre): A PET radioprobe for in vivo tracing of trehalose metabolism.

Trehalose analogues bearing fluorescent and click chemistry tags have been developed as probes of bacterial trehalose metabolism, but these tools have limitations with respect to in vivo imaging applications. Here, we report the radiosynthesis of the 18F-modified trehalose analogue 2-deoxy-2-[18F]fluoro-d-trehalose ([18F]-2-FDTre), which in principle can be used in conjunction with positron emission tomography (PET) imaging to allow in vivo imaging of trehalose metabolism in various contexts. A chemoenzymatic method employing the thermophilic TreT enzyme from Thermoproteus tenax was used to rapidly (15-20 min), efficiently (70% radiochemical yield; ≥ 95% radiochemical purity), and reproducibly convert the commercially available radiotracer 2-deoxy-2-[18F]fluoro-d-glucose ([18F]-2-FDG) into the target radioprobe [18F]-2-FDTre in a single step; both manual and automated syntheses were performed with similar results. Cellular uptake experiments showed that radiosynthetic [18F]-2-FDTre was metabolized by Mycobacterium smegmatis but not by various mammalian cell lines, pointing to the potential future use of this radioprobe for selective PET imaging of infections caused by trehalose-metabolizing bacterial pathogens such as M. tuberculosis.

Single-cell Raman spectroscopy reveals microsporidia spore heterogeneity in various insect hosts.

Single-cell Raman spectroscopy was used to analyze the spore heterogeneity of 16 microsporidia strains from various insect hosts in order to better understand the basic biology of microsporidia. The Raman spectrum of a single spore revealed basic spore composition, and microsporidia spores in various hosts were found to be rich in trehalose. Principal component analysis and Raman intensity showed obvious heterogeneity in the trehalose, nucleic acid, and protein content of various spores; however, there was no correlation between various spore groups and host type. Trehalose content correlated with spore infectivity on Bombyx mori. Raman spectroscopy is an excellent tool for label-free investigation of intercellular molecular constituents, providing insight into the heterogeneity of microsporidia spores.

Screening of Extractant for Determining Active Sludge Trehalose in Sewage Treatment.

This study investigated and proposed the use of trehalose extraction and a detection method for the determining of active sludge trehalose in sewage treatment. Seven extractants (trichloroacetic acid, ethanol, methanol, acetone, pure water, formaldehyde and trichloromethane) were used separately to extract the active sludge trehalose, and their trehalose contents were determined. The results shown in standard curves plotted for all seven extractants demonstrated good linearity, and the regression coefficients varied insignificantly. Using trichloroacetic acid, active sludge trehalose was extracted within a period of only 40 min at 40 centigrade. In view of that, trichloroacetic acid proved to be as the most efficient extractants in extracting trehalose from active sludge. Its extraction rate was 4 to 11-times faster than that of other extractants for the same amount of active sludge. From our results, trichloroacetic acid was substantiated as the optimal extractant for determining active sludge trehalose.