This work aimed to identify the key members of the bacterial community growing on common carp (Cyprinus carpio) fillets during chilled storage with next-generation sequencing (NGS) and cultivation-dependent methods. Carp fillets were stored for 96 h at 2 °C and 6 °C with and without a vacuum package, and an additional frozen-thawed storage experiment was set for 120 days. Community profiles of the initial and stored fish samples were determined by amplicon sequencing. Conventional microbial methods were used parallelly for the enumeration and cultivation of the dominant members of the microbial community. Cultivated bacteria were identified with 16S rRNA sequencing and the MALDI-TOF MS method. Based on our results, the vacuum package greatly affected the diversity and composition of the forming microbial community, while temperature influenced the cell counts and consequently the microbiological criteria for shelf-life of the examined raw fish product. Next-generation sequencing revealed novel members of the chilled flesh microbiota such as Vagococcus vulneris or Rouxiella chamberiensis in the vacuum-packed samples. With traditional cultivation, 161 bacterial strains were isolated and identified at the species level, but the identified bacteria overlapped with only 45% of the dominant operational taxonomic units (OTUs) revealed by NGS. Next-generation sequencing is a promising and highly reliable tool recommended to reach a higher resolution of the forming microbial community of stored fish products. Knowledge of the initial microbial community of the flesh enables further optimization and development of processing and storage technology.
- MeSH
- Bacteria MeSH
- Carps * MeSH
- Microbiota * MeSH
- Seafood microbiology MeSH
- Food Microbiology MeSH
- RNA, Ribosomal, 16S genetics MeSH
- Food Storage methods MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
The sperm of Russian sturgeon (A. gueldenstaedtii) and Siberian sturgeon (A. baerii) was used to evaluate the effects of short-term (liquid) storage on functional parameters (spermatozoa motility and velocity), DNA integrity and oxidative stress indices. Spermatozoa showed >50% motility during 6 days of storage with an average velocity of 133.12±15.4 to 87.9±11.23μm s(-1) in both species. No motile spermatozoa were recorded after nine days of storage. Analysis of Russian sturgeon sperm showed no significant differences in DNA damage expressed as percent tail DNA and Olive Tail Moment for first three days of storage. In Siberian sturgeon significant differences in DNA damage were detected after two days of storage. The level of oxidative stress indices (TBARS, CP) and antioxidant activity (SOD) increased significantly with storage time in both species. Results of this study can be utilized for successful reproduction management and cryopreservation protocols of these endangered species.
- MeSH
- DNA chemistry metabolism MeSH
- Protein Carbonylation MeSH
- Comet Assay veterinary MeSH
- Thiobarbituric Acid Reactive Substances metabolism MeSH
- Sperm Motility physiology MeSH
- Endangered Species MeSH
- Oxidative Stress genetics physiology MeSH
- DNA Damage MeSH
- Fishes genetics physiology MeSH
- Spermatozoa physiology MeSH
- Semen Preservation methods veterinary MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Artificial propagation of sturgeons is becoming increasingly important for recovery efforts as well as for commercial production. Sterlet Acipenser ruthenus is a common Eurasian sturgeon with a small body size and one of the fastest reproductive cycles among the sturgeons. The practical question being addressed in this study was how long fertilization of ovulated eggs can be delayed without substantially reducing the hatching rate, and an ancillary question is under what' temperature conditions do eggs retain good quality. Broodstock were injected with homogenized carp pituitary extract (CPE); ovulated eggs from three females were allocated to various treatment groups for temperature storage (control, 7°C, 11°C, 15°C and 19°C) until fertilized. Storage times at the regulated temperatures prior to fertilization were for 2.5, 5.0, 7.5 and 10.0 h. After the selected storage times in ovarian fluid, eggs were fertilized and transferred to incubation cages and then they were counted. Three replicates were allocated to each storage period and temperature. Hatched larvae were counted at 7-day post-fertilization. We found that sterlet eggs do not need to be fertilized immediately after collection. Reasonably good quality was retained for several hours if temperature conditions are fairly cool and stable. Eggs retained good quality when stored at 7°C and 11°C for up to 10 h with 54.1 ± 2.9 to 69.9 ± 7.9% hatching success, but egg quality was significantly reduced after 5-h storage at 19°C (p < 0.01) and 7.5-h storage at 15°C (p < 0.05) compared to cooler temperatures. Uniform temperatures between 7°C and 11°C can be considered as appropriate for storage of eggs in ovarian fluid for up to 10 h. This information can have practical application to routine hatchery practice for acipenserids, as well as for certain research protocols.
- MeSH
- Time Factors MeSH
- Fertilization MeSH
- Ovum physiology MeSH
- Seasons MeSH
- Reproduction physiology MeSH
- Fisheries MeSH
- Fishes * MeSH
- Temperature * MeSH
- Tissue Preservation methods veterinary MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Bacterial contamination of semen has become an important contributor to the reduced shelf life of insemination doses in the poultry industry, which is why antibiotics (ATBs) are an important component of semen extenders. Due to a global rise in antimicrobial resistance, the aim of this study was to assess the efficiency of selected commercially available semen extenders to prevent possible bacterial contamination of rooster ejaculates. Two selected extenders free from or containing 31.2 μg/mL kanamycin (KAN) were used to process semen samples from 63 healthy Lohmann Brown roosters. Phosphate-buffered saline without ATBs was used as a control. The extended samples were stored at 4 °C for 24 h. Sperm motility, viability, mitochondrial activity, DNA integrity and the oxidative profile of each extended sample were assessed following 2 h and 24 h of storage. Furthermore, selective media were used to quantify the bacterial load and specific bacterial species were identified with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The results indicate that semen extenders enriched with KAN ensured a significantly higher preservation of sperm quality in comparison to their KAN-free counterparts. Bacterial load was significantly decreased in diluents supplemented with ATBs (p ≤ 0.001); however, KAN alone was not effective enough to eradicate all bacteria since several Escherichia coli, Enterococcus faecalis, Enterococcus faecium and Micrococcus luteus were retrieved from samples extended in KAN-supplemented commercial extenders. As such, we may suggest that more focus should be devoted to the selection of an optimal combination and dose of antibiotics for poultry extenders, which should be accompanied by a more frequent bacteriological screening of native as well as extended poultry semen.
- Publication type
- Journal Article MeSH
The aim of our study was to find out the optimal conditions for short-term storage of cerebrospinal fluid (CSF) samples for direct diagnosis of Lyme disease. A mixture of Borrelia-negative CSFs spiked with a defined amount of cultured Borrelia garinii was used. Borrelia stability was investigated over 7 days at four different temperatures [room temperature (RT), +4, -20 and -70 °C]. Quantitative changes in CSF Borrelia were measured by quantitative PCR (qPCR), and morphological changes in the spirochetes were observed by transmission electron microscopy (TEM). These qPCR results were statistically evaluated. We found +4 °C to be an optimal temperature for short-term storage of CSF samples intended for TEM observation. There was no significant difference between the temperatures tested in the average quantity of Borrelia measured by qPCR. On the contrary, electron optical diagnosis of frozen samples and samples stored at RT showed destructive morphological changes and decreased spirochete counts. Our results show that optimal conditions for the pre-analytical phase of investigation of one type of material can differ depending on the diagnostic method employed.
The role of the seminal fluid antioxidant system in protection against damage to spermatozoa during in vitro sperm storage is unclear. This study investigated the effect of in vitro storage of sterlet Acipenser ruthenus spermatozoa together with seminal fluid for 36 h at 4 °C on spermatozoon motility rate and curvilinear velocity, thiobarbituric acid reactive substance level, and components of enzyme and non-enzyme antioxidant system (superoxide dismutase and catalase activity and uric acid concentration) in seminal fluid. Spermatozoon motility parameters after sperm storage were significantly decreased, while the level of thiobarbituric acid reactive substances, activity of superoxide dismutase and catalase, and uric acid concentration did not change. Our findings suggest that the antioxidant system of sterlet seminal fluid is effective in preventing oxidative stress during short-term sperm storage and prompt future investigations of changes in spermatozoon homeostasis and in spermatozoon plasma membrane structure which are other possible reasons of spermatozoon motility deterioration upon sperm storage.
- MeSH
- Catalase metabolism MeSH
- Uric Acid MeSH
- Thiobarbituric Acid Reactive Substances metabolism MeSH
- Sperm Motility MeSH
- Oxidative Stress MeSH
- Fishes physiology MeSH
- Semen metabolism MeSH
- Spermatozoa MeSH
- Superoxide Dismutase metabolism MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
OBJECTIVES: Degradation of coagulation proteins in frozen plasma may influence assay results. The aims of this study were to explore the changes in coagulation parameters in patient plasma and internal quality control (IQC) after different freezing and storage conditions during the short-term and long-term periods. METHODS: Platelet poor plasma was prepared from citrated peripheral blood collected from a group of healthy donors. The plasma was pooled, frozen and stored in a variety of freezing and storage conditions. The changes were monitored using routine coagulation assays, as well as factor VIII (FVIII) and protein S (PS) assays. RESULTS: Plasma stored in liquid nitrogen (LN 2 ) or in -80°C showed long-term stable values for routine tests for a period of over 12 months, and 6 months for FVIII. Interestingly, the activated partial thromboplastin time (aPTT) showed a temporary significant prolongation over the first two weeks. Plasma frozen and stored in -40°C is not viable for aPTT and FVIII testing, otherwise it can be used for other parameters for up to 4 months. PS showed a significant increase in all frozen samples. Freezing rate has a significant impact on plasma quality and the final storage temperature influences the long-term stability. CONCLUSION: The optimal storage conditions are ultra-low temperatures (LN 2 or -80°C) and the highest freezing rate possible. However, frozen plasma is not viable for IQC of aPTT during a period of two weeks after freezing. This study is unique in its conception as a practical guide for the handling of frozen plasma samples in modern laboratory settings.
- MeSH
- Blood Coagulation MeSH
- Hemostatics * MeSH
- Plasma * MeSH
- Humans MeSH
- Partial Thromboplastin Time MeSH
- Blood Coagulation Tests MeSH
- Freezing MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Static ice storage has long been the standard-of-care for lung preservation, although freezing injury limits ischemic time (IT). Controlled hypothermic storage (CHS) at elevated temperature could safely extend IT. This retrospective analysis assesses feasibility and safety of CHS with IT > 15 hours. Three lung transplant (LuTx) centers (April-October 2023) included demographics, storage details, IT, and short-term outcome from 13 LuTx recipients (8 male, 59 years old). Donor lungs were preserved in a portable CHS device at 7 (5-9.3)°C. Indication was overnight bridging and/or long-distance transport. IT of second-implanted lung was 17.3 (15.1-22) hours. LuTx were successful, 4/13 exhibited primary graft dysfunction grade 3 within 72 hours and 0/13 at 72 hours. Post-LuTx mechanical ventilation was 29 (7-442) hours. Intensive care unit stay was 9 (5-28) and hospital stay 30 (16-90) days. Four patients needed postoperative extracorporeal membrane oxygenation (ECMO). One patient died (day 7) following malpositioning of an ECMO cannula. This multicenter experience demonstrates the possibility of safely extending IT > 15 hours by CHS.
- MeSH
- Time Factors MeSH
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Retrospective Studies MeSH
- Aged MeSH
- Cold Ischemia MeSH
- Feasibility Studies MeSH
- Lung Transplantation * methods MeSH
- Organ Preservation * methods MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Multicenter Study MeSH
Storage of carbohydrates in organs protected from disturbance is an important adaptation of plants in disturbed habitats. We carried out a field experiment involving 31 herbaceous plant species in two cultural meadows to find out whether roots or belowground stem-derived organs (stem bases, stem tubers and rhizomes) are the main storage organs, to study how reserves accumulate in individual organs in the long term (growing season) and to ascertain whether meadow abandonment affects the distribution of carbohydrate reserves in plants. We also conducted a 22-day pot experiment with four meadow plant species to determine how removal of roots and aboveground parts affects the use of carbohydrates stored in roots and stem-derived organs in the short term. From the long-term perspective of the field experiment, mowing had a positive effect on the concentration of carbohydrate reserves. From the short-term perspective of the pot experiment, however, the effect on concentration and pools of carbohydrates was negative. In the field experiment, carbohydrate concentrations before winter were generally higher than in mid-season, and more often higher in roots than in stem-derived organs. Roots and stem-derived organs of plants in the pot experiment were depleted similarly after both types of disturbance. Our results indicate a need for including both types of belowground plant organs in future studies of the carbon economy of plants from disturbed habitats.
- MeSH
- Ecosystem * MeSH
- Plant Roots chemistry MeSH
- Poaceae MeSH
- Seasons MeSH
- Plants chemistry MeSH
- Carbohydrates chemistry MeSH
- Plant Stems chemistry MeSH
- Carbon MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Czech Republic MeSH
