scholarly journals Role of Calcium Alginate and Mannitol in Protecting Bifidobacterium

2012 ◽  
Vol 78 (19) ◽  
pp. 6914-6921 ◽  
Author(s):  
Dianawati Dianawati ◽  
Vijay Mishra ◽  
Nagendra P. Shah
Keyword(s):  
Freeze Drying ◽  
Room Temperature ◽  
Cell Envelope ◽  
Aluminum Foil ◽  
Alginate Beads ◽  
Content Type ◽  
Bifidobacterium Animalis ◽  
Freeze Dried ◽  
Cell Envelopes ◽  
Ca Alginate

ABSTRACTFourier transform infrared (FTIR) spectroscopy was carried out to ascertain the mechanism of Ca-alginate and mannitol protection of cell envelope components and secondary proteins ofBifidobacterium animalissubsp.lactisBb12 after freeze-drying and after 10 weeks of storage at room temperature (25°C) at low water activities (aw) of 0.07, 0.1, and 0.2. Preparation of Ca-alginate and Ca-alginate-mannitol as microencapsulants was carried out by dropping an alginate or alginate-mannitol emulsion containing bacteria using a burette into CaCl2solution to obtain Ca-alginate beads and Ca-alginate-mannitol beads, respectively. The wet beads were then freeze-dried. The awof freeze-dried beads was then adjusted to 0.07, 0.1, and 0.2 using saturated salt solutions; controls were prepared by keeping Ca-alginate and Ca-alginate-mannitol in aluminum foil without awadjustment. Mannitol in the Ca-alginate system interacted with cell envelopes during freeze-drying and during storage at low aws. In contrast, Ca-alginate protected cell envelopes after freeze-drying but not during 10-week storage. Unlike Ca-alginate, Ca-alginate-mannitol was effective in retarding the changes in secondary proteins during freeze-drying and during 10 weeks of storage at low aws. It appears that Ca-alginate-mannitol is more effective than Ca-alginate in preserving cell envelopes and proteins after freeze-drying and after 10 weeks of storage at room temperature (25°C).

Scanning Electron Microscopy-cuticular Lesions on the Roundworm Ascaris Suum

1970 ◽  
Vol 28 ◽  
pp. 114-115
Author(s):  
P. A. Madden ◽  
W. R. Anderson
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Freeze Drying ◽  
Room Temperature ◽  
Secondary Electron ◽  
Distilled Water ◽  
Freeze Dried ◽  
Silver Paint ◽  
To Come ◽  
Scanning Electron

The intestinal roundworm of swine is pinkish in color and about the diameter of a lead pencil. Adult worms, taken from parasitized swine, frequently were observed with macroscopic lesions on their cuticule. Those possessing such lesions were rinsed in distilled water, and cylindrical segments of the affected areas were removed. Some of the segments were fixed in buffered formalin before freeze-drying; others were freeze-dried immediately. Initially, specimens were quenched in liquid freon followed by immersion in liquid nitrogen. They were then placed in ampuoles in a freezer at −45C and sublimated by vacuum until dry. After the specimens appeared dry, the freezer was allowed to come to room temperature slowly while the vacuum was maintained. The dried specimens were attached to metal pegs with conductive silver paint and placed in a vacuum evaporator on a rotating tilting stage. They were then coated by evaporating an alloy of 20% palladium and 80% gold to a thickness of approximately 300 A°. The specimens were examined by secondary electron emmission in a scanning electron microscope.

scholarly journals Protective Effects of Tropical Fruit Processing Coproducts on Probiotic Lactobacillus Strains during Freeze-Drying and Storage

2020 ◽  
Vol 8 (1) ◽  
pp. 96 ◽  
Author(s):  
Caroliny Mesquita Araújo ◽  
Karoliny Brito Sampaio ◽  
Francisca Nayara Dantas Duarte Menezes ◽  
Erika Tayse da Cruz Almeida ◽  
Marcos dos Santos Lima ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Freeze Drying ◽  
Room Temperature ◽  
Membrane Integrity ◽  
Added Value ◽  
Protective Effects ◽  
Tropical Fruit ◽  
Freeze Dried ◽  
Fruit Processing ◽  
And Storage

This study evaluated the protective effects of coproducts from agroindustrial processing of the tropical fruits acerola (Malpighia glabra L., ACE), cashew (Anacardium occidentale L., CAS), and guava (Psidium guayaba L., GUA) on the probiotics Lactobacillus paracasei L-10, Lactobacillus casei L-26, and Lactobacillus acidophilus LA-05 during freeze-drying and storage. The occurrence of damage to membrane integrity, membrane potential, and efflux activity of Lactobacillus cells after freeze-drying was evaluated by flow cytometry, and viable counts were measured immediately after freeze-drying and during 90 days of storage under refrigerated or room temperature conditions. Probiotic strains freeze-dried without substrate had the overall highest count reductions (0.5 ± 0.1 to 2.9 ± 0.3 log cycles) after freeze-drying. Probiotics freeze-dried with fruit processing coproducts had small cell subpopulations with damaged efflux activity and membrane potential. Average counts of probiotics freeze-dried with ACE, CAS, or GUA after 90 days of storage under refrigerated or room temperature were in the range of 4.2 ± 0.1 to 5.3 ± 0.2 and 2.6 ± 0.3 to 4.9 ± 0.2 log CFU/g, respectively, which were higher than those observed for strains freeze-dried without substrate. The greatest protective effects on freeze-dried probiotics were overall presented by ACE. These results revealed that ACE, CAS, and GUA can exert protective effects and increase the stability of probiotic lactobacilli during freeze-drying and storage, in addition to supporting a possible added-value destination for these agroindustrial coproducts as vehicles for probiotics and for the development of novel functional foods.

scholarly journals Mycobacterium tuberculosis Rv2700 Contributes to Cell Envelope Integrity and Virulence

2019 ◽  
Vol 201 (19) ◽  
Author(s):  
Edward R. Ballister ◽  
Marie I. Samanovic ◽  
K. Heran Darwin
Keyword(s):  
Growth Rate ◽  
Mycobacterium Tuberculosis ◽  
Cell Envelope ◽  
Bacterial Species ◽  
Transmembrane Helix ◽  
Antibiotic Sensitivity ◽  
Uncharacterized Protein ◽  
Content Type ◽  
Cell Envelopes ◽  
Envelope Functions

ABSTRACT The cell envelope of Mycobacterium tuberculosis is a key target for antibiotics, yet its assembly and maintenance remain incompletely understood. Here we report that Rv2700, a previously uncharacterized M. tuberculosis gene, contributes to envelope integrity. Specifically, an Rv2700 mutant strain had a decreased growth rate, increased sensitivity to antibiotics that target peptidoglycan crosslinking, and increased cell envelope permeability. We propose that Rv2700 be named a “cell envelope integrity” gene (cei). Importantly, a cei mutant had attenuated virulence in mice. Cei shares predicted structural homology with another M. tuberculosis protein, VirR (Rv0431), and we found that a virR mutant had growth rate, antibiotic sensitivity, and envelope permeability phenotypes similar to those of the cei mutant. Both Cei and VirR are predicted to consist of a transmembrane helix and an extracellular LytR_C domain. LytR_C domains have no known function, but they are also found in a family of proteins, the LytR-Cps2A-Psr (LCP) enzymes, that perform important cell envelope functions in a range of bacteria. In mycobacteria, LCP enzymes attach arabinogalactan to peptidoglycan, and mycobacterial LCP enzyme mutants have phenotypes similar to those of virR- and cei-deficient strains. Collectively, our results suggest that LytR_C domain proteins may contribute to the cell envelope functions performed by LCP proteins. This study provides a framework for further mechanistic investigations of LytR_C proteins and, more broadly, for advancing our understanding of the cell envelopes of mycobacteria and other medically and economically important genera. IMPORTANCE Mycobacterium tuberculosis causes about 1.5 million deaths per year. The unique composition of the Mycobacterium tuberculosis cell envelope is required for this bacterium to cause disease and is the target for several critical antibiotics. By better understanding the mechanisms by which mycobacteria assemble and maintain their cell envelope, we might uncover new therapeutic targets. In this work, we show that a previously uncharacterized protein, Rv2700, is important for cell envelope integrity in Mycobacterium tuberculosis and that loss of Rv2700 attenuates virulence in mice. This family of proteins is found in a broad group of bacterial species, so our work provides a first insight into their potential functions in many species important to the environment, industry, and human health.

scholarly journals Environment Shapes the Accessible Daptomycin Resistance Mechanisms in Enterococcus faecium

2019 ◽  
Vol 63 (10) ◽  
Author(s):  
Amy G. Prater ◽  
Heer H. Mehta ◽  
Abigael J. Kosgei ◽  
William R. Miller ◽  
Truc T. Tran ◽  
...  
Keyword(s):  
Enterococcus Faecium ◽  
Cell Envelope ◽  
Multidrug Resistant ◽  
Resistance Mechanisms ◽  
Clinical Strain ◽  
Content Type ◽  
Component Cell ◽  
Evolutionary Trajectories ◽  
High Level ◽  
Cell Envelopes

ABSTRACT Daptomycin binds to bacterial cell membranes and disrupts essential cell envelope processes, leading to cell death. Bacteria respond to daptomycin by altering their cell envelopes to either decrease antibiotic binding to the membrane or by diverting binding away from septal targets. In Enterococcus faecalis, daptomycin resistance is typically coordinated by the three-component cell envelope stress response system, LiaFSR. Here, studying a clinical strain of multidrug-resistant Enterococcus faecium containing alleles associated with activation of the LiaFSR signaling pathway, we found that specific environments selected for different evolutionary trajectories, leading to high-level daptomycin resistance. Planktonic environments favored pathways that increased cell surface charge via yvcRS upregulation of dltABCD and mprF, causing a reduction in daptomycin binding. Alternatively, environments favoring complex structured communities, including biofilms, evolved both diversion and repulsion strategies via divIVA and oatA mutations, respectively. Both environments subsequently converged on cardiolipin synthase (cls) mutations, suggesting the importance of membrane modification across strategies. Our findings indicate that E. faecium can evolve diverse evolutionary trajectories to daptomycin resistance that are shaped by the environment to produce a combination of resistance strategies. The accessibility of multiple and different biochemical pathways simultaneously suggests that the outcome of daptomycin exposure results in a polymorphic population of resistant phenotypes, making E. faecium a recalcitrant nosocomial pathogen.

The effect of heat on the vitamin B6 of milk: II. A comparison of biological and microbiological tests of evaporated milk

1959 ◽  
Vol 26 (2) ◽  
pp. 215-220 ◽  
Author(s):  
Mary K. Davies ◽  
Margaret E. Gregory ◽  
Kathleen M. Henry
Keyword(s):  
Vitamin B6 ◽  
Experimental Error ◽  
Freeze Drying ◽  
Room Temperature ◽  
Raw Milk ◽  
Vitamin B ◽  
Experimental Conditions ◽  
Freeze Dried ◽  
Microbiological Research ◽  
Chick Diet

1. For chicks and rats pyridoxine, pyridoxal and pyridoxamine were equally active in terms of the free bases when given separately from the diet.2. Under our experimental conditions pyridoxine mixed with the chick diet was stable, but 20% of pyridoxamine, and a variable amount of pyridoxal was lost.3. The vitamin B6 activities measured with Saccharomyces carlsbergensis, chicks and rats respectively and expressed as μg. pyridoxine/g. freeze-dried milk were: raw milk 3·4, 3·2 and 4·9; evaporated milk 1·0, 2·1 and 2·7; stored evaporated milk 0·6, 1·4 and 2·0. For the chicks the milks were mixed with the diets; they were given separately to the rats.4. The microbiological and biological results for raw milk agreed within the limits of experimental error. For the processed milks the differences between biological and microbiological tests were statistically significant.5. All three methods of assay showed a 45–70% loss of vitamin B6 activity on processing and a further loss of 30% of the remainder after storage for 6 months at room temperature.We are indebted to Mr J. Rothwell, Department of Dairying, University of Reading, for preparing the evaporated milk and to Dr B. Record, Ministry of Supply, Microbiological Research Establishment, Porton, for freeze-drying the milk. We should like to thank Dr S. K. Kon for his interest in this work.

scholarly journals Freeze-dried plasma proteins are stable at room temperature for at least 1 year

2021 ◽  
Author(s):  
Jaimie Dufresne ◽  
Trung Hoang ◽  
Juliet Ajambo ◽  
Angelique Florentinus-Mefailoski ◽  
Peter Bowden ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Random Sampling ◽  
Freeze Drying ◽  
Room Temperature ◽  
Complement Component ◽  
Separate Experiment ◽  
Plasma Samples ◽  
Esi Ms ◽  
Freeze Dried ◽  
Edta Plasma

Thirty human EDTA plasma samples from male and female subjects ranging in age from 24 to 74 years were collected on ice, processed ice cold and stored frozen at −80 °C, in liquid nitrogen (LN2), or freeze dried and stored at room temperature in a desiccator (FDRT) or freeze dried and stored at −20 °C for 1 year (FD-20). In a separate experiment, EDTA plasma samples were collected onto ice, processed ice cold and maintained on ice ± protease inhibitors versus incubated at room temperature for up to 96 h. Random and independent sampling by liquid chromatography and tandem mass spectrometry (LC–ESI–MS/MS), as correlated by the MASCOT, OMSSA, X!TANDEM and SEQUEST algorithms, showed that tryptic peptides from complement component 4B (C4B) were rapidly released in plasma at room temperature. Random sampling by LC–ESI–MS/MS showed that peptides from C4B were undetectable on ice, but peptides were cleaved from the mature C4B protein including NGFKSHALQLNNR within as little as 1 h at room temperature. The frequency and intensity of precursors within ± 3 m/z of the C4B peptide NGFKSHALQLNNR was confirmed by automated targeted analysis where the precursors from MS/MS spectra that correlated to the target sequence were analyzed in SQL/R. The C4B preproprotein was processed at the N terminus to release the mature chain that was cleaved on the carboxyl side of the isoprene C2 domain within a polar C terminal sequence of the mature C4B protein, to reveal the thioester reaction site, consistent with LC–ESI–MS/MS and Western blot. Random sampling showed that proteolytic peptides from complement component C4B were rarely observed with long term storage at − 80 °C in a freezer or in liquid nitrogen (LN2), freeze drying with storage at − 20 °C (FD-20 °C) or freeze drying and storage at room temperature (FDRT). Plasma samples maintained at room temperature (RT) showed at least 10-fold to 100-fold greater frequency of peptide correlation to C4B and measured peptide intensity compared to samples on ice for up to 72 h or stored at − 80 °C, LN2, FDRT or FD-20 °C for up to a year.

scholarly journals Pengaruh Proses Freeze-Drying dan Penyimpanan pada Suhu Kamar terhadap Viabilitas dan Patogenisitas Plasma Nutfah Mikroba Pasteurella Multocida

2016 ◽  
Vol 12 (1) ◽  
pp. 40
Author(s):  
Siti Chotiah
Keyword(s):  
Freeze Drying ◽  
Room Temperature ◽  
Pasteurella Multocida ◽  
Culture Collection ◽  
Pathogenicity Test ◽  
Drying Process ◽  
Colony Forming Unit ◽  
Freeze Dried ◽  
Conservation Method ◽  
Preservation Medium

<p>The effect of freeze-drying process and preserving in a vacuum at room temperature against viability and pathogenicity of veterinary microbe germ plasma of Pasteuerella multocida BCC 2331 was investigated at Balitvet. The aim of this study was to find out the most effective and efficient conservation method. As much as 5,2 x 1011 colony forming unit (CFU)/ml of bacteria suspension in 7.5% glucose serum as the preservation medium being pathogenic in mice with LD50 of 9,8 CFU/ml was freeze dried then stored at room temperature (&amp;plusmn;27oC) until the study was completed. Viability and pathogenicity test were done immediately after the process, 1 and 2 months after storage. The results showed that there were viability decreases amounted 1,3 x 101 CFU/ml, 102 CFU/ml and 8,2 x l02 CFU/m1 due to the effects of the process, one month and two-month storage respectively. The decreases of pathogenicity on mice were shown by the increases of LD50 amounting log 1, log 2, and log 3 a day after the process, one month and two-month storage respectively.</p><p> </p><p><strong>Abstrak</strong></p><p>Pengaruh proses kering beku dan penyimpanan hasil proses pada suhu kamar 27oC terhadap viabilitas dan patogenisitas plasma nutfah mikroba veteriner telah dipelajari di Balitvet untuk menentukan cara pelestarian yang efektif dan efisien. Dalam kegiatan ini dipakai bakteri Pasteurella multocida koleksi Balitvet Culture Collection nomor koleksi B2331. Suspensi bakteri sebanyak 5,2 x l011 coloni forming unit (CFU)/ml dalam medium preservan 7,5% glukosa, serum dan bersifat patogen pada mencit dengan LD50 9,8 CFU/ml diproses kering beku, kemudian disimpan pada suhu kamar (+27oC) sampai penelitian selesai. Uji viabilitas dan patogenisitas dilakukan langsung setelah proses dan pada 1 serta 2 bulan setelah penyimpanan. Hasil penelitian menunjukkan terjadi penurunan viabilitas sebanyak 1,3 x 101 CFU, dan 8,2 x 102 CFU/ml masingmasing karena pengaruh proses, pengaruh penyimpanan selama 1 dan 2 bulan. Patogenisitas pada mencit menurun yang ditandai oleh adanya peningkatan LD50 sebanyak log 1, log 2, dan log 3 masing-masing 1 hari setelah proses, 1 dan 2 bulan setelah penyimpanan.</p>

scholarly journals Freeze-dried plasma proteins are stable at room temperature for at least 1 year

2021 ◽  
Author(s):  
Jaimie Dufresne ◽  
Trung Hoang ◽  
Juliet Ajambo ◽  
Angelique Florentinus-Mefailoski ◽  
Peter Bowden ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Random Sampling ◽  
Freeze Drying ◽  
Room Temperature ◽  
Complement Component ◽  
Separate Experiment ◽  
Plasma Samples ◽  
Esi Ms ◽  
Freeze Dried ◽  
Edta Plasma

Thirty human EDTA plasma samples from male and female subjects ranging in age from 24 to 74 years were collected on ice, processed ice cold and stored frozen at −80 °C, in liquid nitrogen (LN2), or freeze dried and stored at room temperature in a desiccator (FDRT) or freeze dried and stored at −20 °C for 1 year (FD-20). In a separate experiment, EDTA plasma samples were collected onto ice, processed ice cold and maintained on ice ± protease inhibitors versus incubated at room temperature for up to 96 h. Random and independent sampling by liquid chromatography and tandem mass spectrometry (LC–ESI–MS/MS), as correlated by the MASCOT, OMSSA, X!TANDEM and SEQUEST algorithms, showed that tryptic peptides from complement component 4B (C4B) were rapidly released in plasma at room temperature. Random sampling by LC–ESI–MS/MS showed that peptides from C4B were undetectable on ice, but peptides were cleaved from the mature C4B protein including NGFKSHALQLNNR within as little as 1 h at room temperature. The frequency and intensity of precursors within ± 3 m/z of the C4B peptide NGFKSHALQLNNR was confirmed by automated targeted analysis where the precursors from MS/MS spectra that correlated to the target sequence were analyzed in SQL/R. The C4B preproprotein was processed at the N terminus to release the mature chain that was cleaved on the carboxyl side of the isoprene C2 domain within a polar C terminal sequence of the mature C4B protein, to reveal the thioester reaction site, consistent with LC–ESI–MS/MS and Western blot. Random sampling showed that proteolytic peptides from complement component C4B were rarely observed with long term storage at − 80 °C in a freezer or in liquid nitrogen (LN2), freeze drying with storage at − 20 °C (FD-20 °C) or freeze drying and storage at room temperature (FDRT). Plasma samples maintained at room temperature (RT) showed at least 10-fold to 100-fold greater frequency of peptide correlation to C4B and measured peptide intensity compared to samples on ice for up to 72 h or stored at − 80 °C, LN2, FDRT or FD-20 °C for up to a year.

Sol-Gel Synthesis of Protoenstatite

1994 ◽  
Vol 346 ◽  
Author(s):  
Steven A. Jones ◽  
James M. Burlitch
Keyword(s):  
Hydrogen Peroxide ◽  
Freeze Drying ◽  
Room Temperature ◽  
Sol Gel ◽  
Processing Conditions ◽  
Freeze Dried ◽  
Lower Temperature ◽  
Nmr Methods ◽  
Sol Gel Synthesis ◽  
High Temperature Polymorph

ABSTRACTProtoenstatite, a high-temperature polymorph of enstatite (MgSiO3), is generally not stable at room temperature, and is difficult to synthesize. Using a recently developed, hydrogen peroxide-assisted, sol-gel synthesis, protoenstatite was synthesized in a form that was stable at room temperature. Its crystallization was strongly dependent on processing conditions, particularly on the manner in which the xerogel was formed and fired. Xerogels prepared by evaporation, sprav-drying and freeze-drying were compared by XRD, HTXRD, BET, TG/DTA, and 29Si NMR methods. When samples were prepared by evaporation or spray-drying, the result was a mixture of polymorphs. Only the freeze-dried precursor yielded protoenstatite at a lower temperature and within a shorter time than any previously reported.

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