scholarly journals Effect of Extender, Storage Time and Temperature on Kinetic Parameters (CASA) on Bull Semen Samples

Biology ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 806
Author(s):  
Aitor Fernandez-Novo ◽  
Sergio Santos-Lopez ◽  
Clara Barrajon-Masa ◽  
Patricia Mozas ◽  
Eduardo de Mercado ◽  
...  

CASA kinetic parameters are often evaluated in a diagnostic centre. How storage conditions affect ejaculates up to evaluation is unclear. We assessed, in 25 commercial bulls electroejaculated in the field, the impact of time until evaluation (0–2 h, 4–6 h, and 24 h post-ejaculation), holding temperature (5 °C vs. room temperature), and extender (AndroMed®, BIOXcell® or INRA96®) on CASA kinetic parameters. Total and progressive motility, VCL, VAP, VCL, ALH, BCF, STR, LIN, and WOB were assessed. CASA kinetic parameters were preserved for up to 4–6 h post-ejaculation, except for AndroMed®. Regardless of extender or temperature, motility decreased from 4–6 h up to 24 h, with the best values obtained with BIOXcell® at 5 °C. Our results suggest that BIOXcell® can preserve sperm motility for up to 6 h, either at 5 °C or room temperature, and also INRA96® at room temperature, with motility assessments and the percentage of the most rapid sperms being the lowest with INRA96® at 5 °C. The kinetic parameters decreased when analyses were performed at 24 h. Therefore, we suggest evaluating seminal quality as soon as possible, before 6 h after collection. These results help to fix adequate protocols for the short-term storage and shipment of bovine semen collected under field conditions.

Biology ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 630
Author(s):  
Aitor Fernandez-Novo ◽  
Sergio Santos-Lopez ◽  
Clara Barrajon-Masa ◽  
Patricia Mozas ◽  
Eduardo de Mercado ◽  
...  

Seminal parameters can be evaluated in situ, or samples can be delivered to a diagnostic centre. How storage conditions affect ejaculates up to evaluation is unclear. We assessed, in 25 commercial bulls electroejaculated in the field, the impact of time until evaluation (0–2 h, 4–6 h, and 24 h post-ejaculation), holding temperature (5 °C vs. room temperature), and extender (AndroMed®, BIOXcell® or INRA96®) on semen quality. Acrosome integrity, sperm viability and morphology, CASA-total and progressive motility, pH, and colony-forming units were assessed. Semen quality was preserved for up to 4–6 h post-ejaculation, except for INRA96® at 5 °C. Regardless of extender or temperature, motility decreased from 4 to 6 h up to 24 h, with the best values obtained with BIOXcell® at 5 °C. pH differed from 4 to 6 h up to 24 h, acidifying when stored at room temperature. Microbiological load was stable over time with AndroMed® and BIOXcell®, and increased at room temperature with INRA96®. Our results suggest that AndroMed® and BIOXcell® can preserve semen quality for up to 6 h, either at 5 °C or room temperature, while INRA96® only at room temperature. These results help to fix adequate protocols for short-term storage and shipment of bovine semen collected under field conditions.


Water ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 1321 ◽  
Author(s):  
Sergi Astals ◽  
Konrad Koch ◽  
Sören Weinrich ◽  
Sasha D. Hafner ◽  
Stephan Tait ◽  
...  

The impact of storage temperature (4, 22 and 37 °C) and storage time (7, 14 and 21 days) on anaerobic digestion inocula was investigated through specific methanogenic activity assays. Experimental results showed that methanogenic activity decreased over time with storage, regardless of storage temperature. However, the rate at which the methanogenic activity decreased was two and five times slower at 4 °C than at 22 and 37 °C, respectively. The inoculum stored at 4 °C and room temperature (22 °C) maintained methanogenic activity close to that of fresh inoculum for 14 days (<10% difference). However, a storage temperature of 4 °C is preferred because of the slower decrease in activity with lengthier storage time. From this research, it was concluded that inoculum storage time should generally be kept to a minimum, but that storage at 4 °C could help maintain methanogenic activity for longer.


Foods ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 968
Author(s):  
David A. Vargas ◽  
Markus F. Miller ◽  
Dale R. Woerner ◽  
Alejandro Echeverry

The use of antimicrobials in the pork industry is critical in order to ensure food safety and, at the same time, extend shelf life. The objective of the study was to determine the impact of antimicrobials on indicator bacteria on pork loins under long, dark, refrigerated storage conditions. Fresh boneless pork loins (n = 36) were split in five sections and treated with antimicrobials: Water (WAT), Bovibrom 225 ppm (BB225), Bovibrom 500 ppm (BB500), Fit Fresh 3 ppm (FF3), or Washing Solution 750 ppm (WS750). Sections were stored for 1, 14, 28, and 42 days at 2–4 °C. Mesophilic and psychrotrophic aerobic bacteria (APC-M, APC-P), lactic acid bacteria (LAB-M), coliforms, and Escherichia coli were enumerated before intervention, after intervention, and at each storage time. All bacterial enumeration data were converted into log10 for statistical analysis, and the Kruskal–Wallis test was used to find statistical differences (p < 0.05). Initial counts did not differ between treatments, while, after treatment interventions, treatment WS750 did not effectively reduce counts for APC-M, APC-P, and coliforms (p < 0.01). BB500, FF3, and WS750 performed better at inhibiting the growth of indicator bacteria when compared with water until 14 days of dark storage.


2021 ◽  
pp. 108201322199884
Author(s):  
Rami Akkad ◽  
Ereddad Kharraz ◽  
Jay Han ◽  
James D House ◽  
Jonathan M Curtis

The odour emitted from the high-tannin fab bean flour ( Vicia faba var. minor), was characterized by headspace solid-phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC–MS). The relative odour activity value (ROAV) was used to monitor the changes in key volatile compounds in the flour during short-term storage at different temperature conditions. The key flavour compounds of freshly milled flour included hexanal, octanal, nonanal, decanal, 3-methylbutanal, phenyl acetaldehyde, (E)-2-nonenal, 1-hexanol, phenyl ethyl alcohol, 1-octen-3-ol, β-linalool, acetic acid, octanoic acid, and 3-methylbutyric acid; these are oxidative degradation products of unsaturated fatty acids and amino acids. Despite the low lipid content of faba beans, the abundances of aldehydes arising during room temperature storage greatly contributed to the flavour of the flour due to their very low odour thresholds. Two of the key volatiles responsible for beany flavour in flour (hexanal, nonanal) increased greatly after 2 weeks of storage at room temperature or under refrigerated conditions. These volatile oxidation products may arise as a result of enzymatic activity on unsaturated fatty acids, and was seen to be arrested by freezing the flour.


Forests ◽  
2021 ◽  
Vol 12 (5) ◽  
pp. 528
Author(s):  
Jelena Kranjec Orlović ◽  
Damir Drvodelić ◽  
Marko Vukelić ◽  
Matea Rukavina ◽  
Danko Diminić ◽  
...  

When natural regeneration of Quercus robur stands is hampered by an insufficient acorn yield, human assisted sowing of acorns collected in non-affected stands and stored for some period of time is performed. To inhibit the development of fungi and acorn deterioration during storage, thermotherapy is usually applied by submerging acorns for 2.5 h in water heated to 41 °C. This research aimed to test the effect of four thermotherapy treatments of different durations and/or applied temperatures as well as short-term storage at −1 °C or 3 °C on acorn internal mycobiota and germination. Fungal presence in cotyledons was analyzed in 450 acorns by isolation of mycelia on artificial media, followed by a DNA-based identification. Germination of 2000 acorns was monitored in an open field trial. Thermotherapy significantly decreased fungal diversity, while storage at 3 °C increased the isolation frequency of several fungi, mainly Penicillium spp. The most frequently isolated fungi did not show a negative impact on acorn germination after short-term storage. The study confirmed the efficiency of thermotherapy in the eradication of a part of acorn internal mycobiota, but also its effect on the proliferation of fast-colonizing fungi during storage. However, the latter showed to be more stimulated by storage conditions, specifically by storage at 3 °C.


Metabolites ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 515
Author(s):  
Daniela Duarte ◽  
Beatriz Castro ◽  
Joana Leonor Pereira ◽  
Joana Faria Marques ◽  
Ana Luísa Costa ◽  
...  

Maintaining a salivary metabolic profile upon sample collection and preparation is determinant in metabolomics. Nuclear magnetic resonance (NMR) spectroscopy was used to identify metabolite changes during short-term storage, at room temperature (RT)/4 °C/−20 °C, and after sample preparation, at RT/4 °C (mimicking typical clinical/laboratory settings). Interestingly, significant metabolic inter-individual and inter-day variability were noted, probably determining sample stability to some extent. After collection, no changes were noted at −20 °C (at least for 4 weeks). RT storage induced decreases in methylated macromolecules (6 h); lactate (8 h); alanine (12 h); galactose, hypoxanthine, pyruvate (24 h); sarcosine, betaine, choline, N-acetyl-glycoproteins (48 h), while acetate increased (48 h). Less, but different, changes were observed at 4 °C, suggesting different oral and microbial status at different temperatures (with a possible contribution from inter-individual and inter-day variability), and identifying galactose, hypoxanthine, and possibly, choline esters, as potential general stability indicators. After preparation, addition of NaN3 did not impact significantly on saliva stabilization, neither at RT nor at 4 °C, although its absence was accompanied by slight increases in fucose (6.5 h) and proline (8 h) at RT, and in xylose (24 h) at 4 °C. The putative metabolic origins of the above variations are discussed, with basis on the salivary microbiome. In summary, after collection, saliva can be stored at RT/4 °C for up to 6 h and at −20 °C for at least 4 weeks. Upon preparation for NMR analysis, samples are highly stable at 25 °C up to 8 h and at 4 °C up to 48 h, with NaN3 addition preventing possible early changes in fucose, proline (6–8 h), and xylose (24 h) levels.


2020 ◽  
Vol 100 (10) ◽  
pp. 1345-1355 ◽  
Author(s):  
Stefaniya Boneva ◽  
Anja Schlecht ◽  
Daniel Böhringer ◽  
Hans Mittelviefhaus ◽  
Thomas Reinhard ◽  
...  

Abstract This study aims to compare the potential of standard RNA-sequencing (RNA-Seq) and 3′ massive analysis of c-DNA ends (MACE) RNA-sequencing for the analysis of fresh tissue and describes transcriptome profiling of formalin-fixed paraffin-embedded (FFPE) archival human samples by MACE. To compare MACE to standard RNA-Seq on fresh tissue, four healthy conjunctiva from four subjects were collected during vitreoretinal surgery, halved and immediately transferred to RNA lysis buffer without prior fixation and then processed for either standard RNA-Seq or MACE RNA-Seq analysis. To assess the impact of FFPE preparation on MACE, a third part was fixed in formalin and processed for paraffin embedding, and its transcriptional profile was compared with the unfixed specimens analyzed by MACE. To investigate the impact of FFPE storage time on MACE results, 24 FFPE-treated conjunctival samples from 24 patients were analyzed as well. Nineteen thousand six hundred fifty-nine transcribed genes were detected by both MACE and standard RNA-Seq on fresh tissue, while 3251 and 2213 transcripts were identified explicitly by MACE or RNA-Seq, respectively. Standard RNA-Seq tended to yield longer detected transcripts more often than MACE technology despite normalization, indicating that the MACE technology is less susceptible to a length bias. FFPE processing revealed negligible effects on MACE sequencing results. Several quality-control measurements showed that long-term storage in paraffin did not decrease the diversity of MACE libraries. We noted a nonlinear relation between storage time and the number of raw reads with an accelerated decrease within the first 1000 days in paraffin, while the numbers remained relatively stable in older samples. Interestingly, the number of transcribed genes detected was independent on FFPE storage time. RNA of sufficient quality and quantity can be extracted from FFPE samples to obtain comprehensive transcriptome profiling using MACE technology. We thus present MACE as a novel opportunity for utilizing FFPE samples stored in histological archives.


2016 ◽  
Vol 79 (5) ◽  
pp. 816-820
Author(s):  
LARISSA NAZARETH de FREITAS ◽  
LAERTE DAGHER CASSOLI ◽  
JANIELEN da SILVA ◽  
JOSÉ CARLOS de FIGUEIREDO PANTOJA ◽  
PAULO FERNANDO MACHADO

ABSTRACT Total bacterial count (TBC) is a tool used to assess milk quality and is associated with not only the initial sample contamination but also the sample storage time and temperature. Several countries have reported milk samples with a high TBC, and the influence of TBC on milk preservation remains unclear. Thus, the aim of this study was to evaluate the impact of the initial bacterial contamination level on the macrocomponents and somatic cell count (SCC) of raw milk samples preserved with bronopol and maintained at two storage temperatures (7 and 25°C) for up to 12 days. Thus, we collected milk samples from 51 dairy farms, which were divided into two groups according to the initial bacterial load: low TBC (&lt;100,000 CFU/ml) and high TBC (≥100,000 CFU/ml). We analyzed the sample composition for protein, fat, total solids, lactose, milk urea nitrogen, and the SCC. We did not observe an effect from TBC and storage time and temperature on the concentration of protein, fat, total solids, and lactose. SCC changes were not observed for samples maintained under refrigeration (7°C); however, samples maintained at room temperature (25°C) exhibited a decrease in the SCC beginning on day 6 of storage. For milk urea nitrogen, values increased when the samples were maintained at room temperature, beginning on the ninth storage day. Samples with the preservative bronopol added and maintained under refrigeration may be analyzed up to 12 days after collection, regardless of the milk microbial load.


2004 ◽  
Vol 94 (4) ◽  
pp. 353-355 ◽  
Author(s):  
Anthony Rosales ◽  
Jefferey R. Vazquez ◽  
Brian Short ◽  
Heather R. Kimbriel ◽  
Matthew J. Claxton ◽  
...  

Maggot debridement therapy is rapidly increasing in popularity at major diabetic foot and wound care centers worldwide. However, we are unaware of specific guidelines on the short-term storage of larvae. We sought to evaluate differences in maggot motility over time in larvae refrigerated versus those stored at room temperature. We also introduce a simple surrogate method for evaluating maggot vitality that may be useful for in vivo studies if validated in future works. We randomly selected ten larvae from the same shipment at ten different times in 9 days. Larvae were placed on a translucent acetate grid, and their total excursion in 30 sec was measured. This was converted into a Maggot Motility Index. In the refrigerated group, the index remained at or above 40 mm/min for approximately 60 hours from baseline, when there was a significant decrease. This same phenomenon occurred during the first 12 hours in the nonrefrigerated group. There were significant differences in motility between refrigerated and nonrefrigerated larvae immediately after baseline until day 8. Larvae are more practical for repeated clinical use if kept refrigerated between applications. (J Am Podiatr Med Assoc 94(4): 353–355, 2004)


2019 ◽  
Vol 51 (2) ◽  
pp. 159-163
Author(s):  
B. Alev ◽  
S. Tunali ◽  
R. Yanardag ◽  
A. Yarat

Enzymes are made of protein, that is why they are sensitive molecules and are affected by storage conditions. A small change in enzyme activity during storage may cause a big error in analysis results. The aim of the study was to evaluate the effects of storage time and temperature on urease activity. Urease solutions were prepared at different activities (from 100 to 2000 U/mL) and stored at room temperature, in the refrigerator (4°C), and in the deep freezer (-18°C and -80°C). Activity measurements were made at regular intervals until 28 days by the modified Weatherburn method. The relative activities of 100-1000 U/mL urease solutions stored at room temperature, 4, -18 or -80°C were 75% and below after 4 days. Twenty-eight days later, for 2000 U/mL urease solutions, only at room temperature, the relative activity was reduced to 37%, while at 4, -18 or -80°C, the relative activities were above 80%. Since urease can be maintained at 4°C for 28 days without significant loss of activity, it has practical importance. Low-activity urease solutions (such as 100-1000 U/mL) should not be stored at -18 or -80°C for short or long term storage, they should be stored at 4°C only for one day. Keywords: Urease activity, storage time, storage temperature


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