138 Associations of sperm head morphometrics with quality parameters of frozen-thawed ram semen

2020 ◽  
Vol 32 (2) ◽  
pp. 195
Author(s):  
J. Navaranjan ◽  
J. Szymanowicz ◽  
M. Murawski ◽  
T. Schwarz ◽  
P. M. Bartlewski
Keyword(s):  
Alkaline Phosphatase ◽  
Survival Time ◽  
Aspartate Aminotransferase ◽  
Sperm Quality ◽  
Sperm Head ◽  
Head Length ◽  
Cell Membrane Permeability ◽  
Sperm Length ◽  
Progressive Motility ◽  
Synthetic Line

Current methods of mammalian semen evaluation focus on determining spermatozoa motility, concentration, mitochondrial status, and nucleus or chromatin structure integrity, quantifying their ability to bind to ova or measuring seminal plasma content of various biochemical markers. However, there is a paucity of studies that address relationships between sperm head morphometry (the external shape and dimensions of the sperm) and fertilising ability. Sperm head morphometrics are influenced by many molecular and biochemical factors such as genetics, DNA or protein condensation, and cell membrane permeability, all of which can affect semen viability. The objective of this experimental work was to determine quantitative correlations between sperm head dimensions and various indices of sperm quality in frozen-thawed ram semen. Ejaculates were collected from 16 clinically healthy rams (4 Polish Lowland (PON), 4 Olkuska, 5 synthetic line BCP (Berrichon du Cher×Charolais×PON/Polish Merino), and 3 synthetic line SCP (Suffolk×Charolais×PON/Polish Merino) aged 4-12 years) into an artificial vagina in the middle portion of the breeding season. Ejaculates from each ram were divided into two equal portions, diluted with a commercial semen extender prepared in deionised water or nanowater (water declusterised using cold plasma treatment) to a final concentration of 400×106 spermatozoamL−1, and frozen in 0.25-mL plastic straws. After 6 months of being cryogenically preserved, semen samples were thawed and used for the preparation of smears stained with eosin or SpermBlue. Images of the samples containing at least 100 spermatozoa were taken under 200× magnification and used for determination of sperm head morphology with the image analytical software Image Pro Plus (Media Cybernetics Inc.). Sperm progressive motility and survival time, as well as extender concentrations of alkaline phosphatase and aspartate aminotransferase, were measured. Finally, 128 BCP ewes were inseminated laparoscopically with the ram semen and fertility parameters were recorded. The present data were analysed using a multivariate analysis of variance in SAS (SAS Institute Inc.) and Spearman correlation tests. There were no significant effects or interactions of breed, staining method, or extender diluent on sperm head dimensions (head length, width, area, perimeter, and roundness). The mean head length was negatively correlated (P<0.05) with the percentages of spermatozoa with vacuolated, detached, or amorphous heads or small acrosomes; thick and thin midpiece defects, distal droplet, broken tail plus distal droplet, short tail plus distal droplet, and thick midpiece plus proximal droplet; and sperm progressive motility. In addition, sperm head roundness was negatively correlated with the proportion of spermatozoa with coiled tails. There were no correlations of sperm head dimensions with survival time, alkaline phosphatase and aspartate aminotransferase concentrations, or conception and pregnancy rates of artificially inseminated ewes. Sperm length and roundness (but no other measurements) were significantly correlated with segmental sperm defects and motility that may impinge the fertilising ability of frozen-thawed ram semen.

scholarly journals Fertilization Modes And The Evolution of Sperm Characteristics In Marine Fishes: Paired Comparisons of The Externally And Internally Fertilizing Species

2021 ◽  
Author(s):  
Takeshi Ito ◽  
Masaya Morita ◽  
Kazuo Inaba ◽  
Kogiku Shiba ◽  
Hiroyuki Munehara ◽  
...  
Keyword(s):  
Sperm Competition ◽  
Sperm Motility ◽  
Sperm Head ◽  
Marine Fishes ◽  
Head Length ◽  
Sperm Length ◽  
Sperm Velocity ◽  
Internal Fertilization ◽  
Sperm Characteristics ◽  
Testis Mass

Abstract Background: Fertilization modes may affect sperm characteristics, such as morphology, velocity, and motility. However, there is surprisingly little information on how fertilization mode affects sperm evolution because several factors (e.g. sperm competition and phylogeny) are intricately intertwined with this factor when phylogenetically distant species are compared. Here, we compared sperm characteristics between six externally and four internally fertilizing marine fishes from three different groups containing close relatives, taking into account the level of sperm competition. We also analysed the relationship between relative testis mass (as an index of sperm competition level) and sperm characteristics.Results: Sperm head morphology was significantly longer in species with internal fertilization than in those with external fertilization, suggesting that a longer head is advantageous for swimming in viscous ovarian fluid or the complex ovarian structure. In addition, sperm motility differed between external fertilizers and internal fertilizers; sperm of externally fertilizing species were only motile in seawater, and sperm of internally fertilizing species were only motile in an isotonic solution. These results suggest that sperm motility has adapted according to the fertilization mode. In contrast, total sperm length and sperm velocity did not correlate with fertilization mode, perhaps because of the different levels of sperm competition. Relative testis mass is positively correlated with sperm velocity and negatively correlated with the ratio of sperm head length/flagellum length. This finding suggests that species with high levels of sperm competition have sperm that are fast and have relatively long flagella compared to head length. These results contradict a previous assumption that the evolution of internal fertilization increases total sperm length. In addition, copulatory behaviour with internal insemination may involve a large intromittent organ, but this is not essential in fish, probably due to the avoidance of water resistance.Conclusions: We propose a new scenario of sperm evolution in which internal fertilization increases sperm head length but not total sperm length and changes sperm motility. In contrast, sperm competition affects sperm length and velocity. Our findings provide a new perspective on the evolutionary biology of sperm in fish.

scholarly journals Proteins of the canine seminal plasma

2016 ◽  
Vol 46 (5) ◽  
pp. 901-908 ◽  
Author(s):  
Annice Aquino-Cortez ◽  
Lúcia Daniel Machado da Silva ◽  
Airton Alencar de Araújo ◽  
Erika da Silva Bezerra de Menezes ◽  
Arlindo de Alencar Araripe Noronha Moura
Keyword(s):  
Alkaline Phosphatase ◽  
Seminal Plasma ◽  
Binding Proteins ◽  
Reproductive Tract ◽  
Sperm Quality ◽  
Zinc Binding ◽  
Heparin Binding ◽  
Reproductive Disorders ◽  
The Matrix ◽  
Peroxidase Enzymes

ABSTRACT: Studies have been performed to identify the proteins present in canine seminal plasma (SP) and relate them to sperm quality as well as to discover molecular markers of reproductive tract diseases. There is evidence that heparin-binding proteins, zinc-binding proteins, and lactoferrin as well as the matrix metalloproteinase, superoxide dismutase, catalase, and glutathione peroxidase enzymes are associated with canine sperm quality. Other studies indicate that prolactin and enzymes like arginine esterase, acid phosphatase, and alkaline phosphatase could be successfully used as biomarkers of reproductive disorders. Thus, the present literature review aims to address aspects related to proteins of the canine SP, their influence on fertility, and their importance as biomarkers of reproductive disorders.

Hepatotoxic effects of chloroform extract of Artemisia macivera Linn in rats following intraperitoneal administration of different subchronic doses

2010 ◽  
Vol 30 (1) ◽  
pp. 25-33 ◽  
Author(s):  
SE Atawodi ◽  
AC Ene ◽  
DA Ameh
Keyword(s):  
Alkaline Phosphatase ◽  
Total Protein ◽  
Aspartate Aminotransferase ◽  
Alanine Aminotransferase ◽  
Intraperitoneal Administration ◽  
Chloroform Extract ◽  
Hepatic Tissue ◽  
Albino Rats ◽  
High Dose ◽  
Hepatocellular Injury

The possible hepatotoxic effects of chloroform extract of Artemisia maciverae was evaluated biochemically and histologically using male Swiss albino rats, randomly assigned into four groups of 24 animals each. The groups (control, 50, 100 and 200 mg/kg body weight) were treated for 60 days and then monitored for another 30 days before sacrifice. Alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, bilirubin (total and direct), total protein and albumin were assessed colorimetrically, while tissue specimens were subjected to histological examination following standard hematoxyline-eosin staining techniques. After 1 week of treatment, the extract caused statistically significant elevation in levels of serum alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and bilirubin (total and direct), while there was significant (p < 0.05) decrease in the levels of serum total protein and albumin at the onset of treatment when compared with the control. These abnormalities in the levels of serum biochemical parameters were spontaneously corrected within 2 weeks of treatment. Similarly, histological assessment showed severe hepatic tissue injuries after 1 week, but these organs recovered spontaneously by the second week of treatment. The results indicate that long-term exposure to therapeutic doses of chloroform extract of A maciverae is relatively safe, but high dose exposure may result in hepatocellular injury.

scholarly journals Morphometric Characteristics of the Spermatozoa of Blue Fox (Alopex lagopus) and Silver Fox (Vulpes vulpes)

Animals ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 1927
Author(s):  
Katarzyna Andraszek ◽  
Dorota Banaszewska ◽  
Olga Szeleszczuk ◽  
Marta Kuchta-Gładysz ◽  
Anna Grzesiakowska
Keyword(s):  
Vulpes Vulpes ◽  
Tail Length ◽  
Head Length ◽  
Head Width ◽  
Sperm Length ◽  
Alopex Lagopus ◽  
Silver Fox ◽  
Shape Indices ◽  
Length Width ◽  
Blue Fox

The results presented in this study are the first such extensive characterization of the sperm morphometry of the blue fox (Alopex lagopus) and silver fox (Vulpes vulpes), as representatives of the family Canidae. Canine spermatozoa, especially the sperm of farmed foxes, are not often described in studies on reproduction. The aim of the study was a detailed comparison of the morphometric dimensions and shape of the sperm of two fox species: silver fox and blue fox. Semen collected from 10 silver foxes and 10 blue foxes was used for the study. The specimens were stained with silver nitrate. Measurements were performed of the length, width, perimeter, and area of the head; the area of the acrosome and its coverage; the length of the midpiece and its coverage; the length of the tail; and the length of the end piece of the tail. In addition, four head shape indices were calculated: ellipticity, elongation, roughness and regularity. The following values for the morphometric parameters and shape indices were obtained for blue fox and silver fox, respectively: head length—6.72 µm and 6.33 µm; head width—4.54.µm and 4.21 µm; head perimeter—18.11 µm and 17.37 µm; head area—21.94 µm2 and 21.11 µm2; acrosome area—11.50 µm2 and 10.92 µm2; midpiece length—12.85 µm and 12.79 µm; tail end piece length—3.44 µm and 3.28 µm; tail length—65.23 µm and 65.09 µm; acrosome coverage—52.43% and 52.83%; midpiece coverage—19.71% and 19.65%; sperm length—71.95 µm and 71.42 µm; ellipticity—1.49 and 1.52; elongation—0.19 and 0.20; roughness—0.84 and 1.88; regularity—1.09 and 0.99. The significance of differences between species was verified by Tukey’s test at p ≤ 0.05. Statistically significant differences between species were found for the following parameters: head length, width, perimeter and area; acrosome area; tail, end piece, and total sperm length; roughness and regularity. The differences in the size and shape of sperm can be used to establish reference patterns for fox sperm enabling more accurate species identification.

scholarly journals Comparison of a full-spectrum multi-analyte clinical analyser with six reference instruments using canine and feline blood samples

2017 ◽  
Vol 62 (No. 6) ◽  
pp. 342-350
Author(s):  
CS Lin ◽  
GH Chiang ◽  
CH Liu ◽  
HC Tsai ◽  
CC Yang ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Bile Acid ◽  
Blood Urea Nitrogen ◽  
Total Protein ◽  
Aspartate Aminotransferase ◽  
Alanine Aminotransferase ◽  
Total Bilirubin ◽  
Total Bile Acid ◽  
Analytical Performance ◽  
Blood Samples

In this study, we report the characterisation of a novel centrifugation and spectrum-integrated veterinary clinical analyser, the AmiShield<sup>TM</sup>, which has been developed for the multiplex measurement of biochemical, electrolyte and immunoassay parameters in a point-of-care testing environment. The aims of this study were to evaluate the analytical performance of the AmiShield<sup>TM</sup> and to compare it with six reference instruments using clinical blood samples. Two hundred and four canine and 120 feline blood samples collected from veterinary teaching hospitals were analysed in parallel using the AmiShield and appropriate reference instruments. All results were evaluated separately for canine and feline specimens. The instrument’s analytical performance was evaluated initially for short- and long-term precision, bias, and observed total error using quality control material. This was followed by comparison of clinical specimens on the AmiShield analyser in parallel with the Vitros and Hitachi for biochemical parameters, VetScan and SNAPshot for total bile acids, and VetLyte and Biolyte for electrolytes. Overall, the AmiShield analyser’s performance met the standards of the American Society for Veterinary Clinical Pathology for total allowable error for most analytes, and can be considered suitable for use in veterinary clinical practices. Using canine samples, excellent correlation coefficients (r ≧ 0.92) were identified for 14 analytes of various categories including glucose, total protein, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, total bilirubin, amylase, blood urea nitrogen, creatinine, phosphorus, Na<sup>+</sup>, K<sup>+</sup>, Cl<sup>–</sup> and total bile acid, while good correlations (0.91 ≧ r ≧ 0.80) were recorded for albumin (r = 0.91). Bland-Altman difference plots also showed agreement (greater than 95% within Limits of Agreement) for glucose, total protein, albumin, alanine aminotransferase, alkaline phosphatase, total bilirubin, amylase, blood urea nitrogen, creatinine, Na<sup>+</sup>, K<sup>+</sup>, Cl<sup>–</sup> and total bile acid between AmiShield and the reference instruments. However, aspartate aminotransferase and phosphorus exhibited higher outliers, implying potential problems associated with matrix interferences such as lipemic samples, which warrant further study. This study demonstrates that the AmiShield compares favourably with standard reference instruments, and the new device generated data of high quality for most analytes in clinical canine and feline samples. The capability of reliably measuring multi-category analytes in one device using minute amounts (170 μl) of whole blood and short turn-around times (&lt; 15 min) underlines the high potential of the device as a good alternative in-house diagnostic application.

scholarly journals Effect of Various Diluents on the Activity of Several Enzymes Present in Serum

1973 ◽  
Vol 19 (9) ◽  
pp. 1079-1080
Author(s):  
Ted W Fendley ◽  
Jane M Hochholzer ◽  
Christopher S Frings
Keyword(s):  
Alkaline Phosphatase ◽  
Creatine Kinase ◽  
Enzyme Activity ◽  
Lactate Dehydrogenase ◽  
Confidence Level ◽  
Aspartate Aminotransferase ◽  
Nacl Solution ◽  
Manual Method ◽  
Accuracy And Precision ◽  
Significant Difference

Abstract We have evaluated the effect of diluting serum with water or NaCl solution (8.5 or 9.0 g/liter) before assaying by a manual method for creatine kinase (EC 2.7.3.2), alkaline phosphatase (EC 3.1.3.1), lactate dehydrogenase (EC 1.1.1.27), and aspartate aminotransferase (EC 2.6.1.1) activity. The t test and the F test show no significant difference in the accuracy and precision of the assays at the 95% confidence level when 100 different samples were compared for each enzyme activity after use of the three diluents.

scholarly journals Finding an Effective Freezing Protocol for Turkey Semen: Benefits of Ficoll as Non-Permeant Cryoprotectant and 1:4 as Dilution Rate

Animals ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 421 ◽  
Author(s):  
Michele Di Iorio ◽  
Giusy Rusco ◽  
Roberta Iampietro ◽  
Maria Antonietta Colonna ◽  
Luisa Zaniboni ◽  
...  
Keyword(s):  
Dilution Rate ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Quality Parameters ◽  
Dna Integrity ◽  
Osmotic Resistance ◽  
Progressive Motility ◽  
Nitrogen Vapor ◽  
Combined Use ◽  
Cryopreservation Protocol

The present study aimed to find an effective cryopreservation protocol for turkey semen through the combined use of dimethylsulfoxide (DMSO) and three non-permeant cryoprotectants (NP-CPAs), sucrose, trehalose, and Ficoll 70. In addition, the action of two dilution rates (1:2 and 1:4) were also investigated. Semen was processed according to two final dilution rates and the following treatments: Tselutin extender (TE)/DMSO (control), TE/DMSO + sucrose or trehalose 50, 100, 200, or 400 mM, and TE/DMSO + Ficoll 0.5, 0.75, 1, or 1.5 mM. In total 26 different combinations treatments were achieved. The diluted semen was filled up into straws and frozen on liquid nitrogen vapor. The post-thawing sperm quality was assessed by analyzing motility, membrane integrity, osmotic resistance, and DNA integrity. The results obtained revealed a significant effect of NP-CPA concentration on total and progressive motility, on most of the kinetic parameters, on membrane integrity and DNA integrity, while the post-thaw quality was less affected by dilution rate. The highest post-thaw quality for all sperm quality parameters assessed except curvilinear velocity (VCL) and DNA integrity were found in semen frozen with 1 mM Ficoll/1:4 (p < 0.05). Our findings provide an important contribution for the identification of a reference procedure for turkey semen cryopreservation, in order to create the first national avian semen cryobank.

Some Methodological Modifications of the Technicon "SMA 12/60 AutoAnalyzer" System

1972 ◽  
Vol 18 (7) ◽  
pp. 630-642 ◽  
Author(s):  
Tevfik K Bigat ◽  
Abraham Saifer
Keyword(s):  
Alkaline Phosphatase ◽  
Glucose Oxidase ◽  
Aspartate Aminotransferase ◽  
Inorganic Phosphate ◽  
New Method ◽  
Green Method ◽  
Bromcresol Green ◽  
Made In

Abstract Methodological modifications have been made in our "SMA 12/60 AutoAnalyzer" system, to improve the reproducibility and accuracy of six commonly performed procedures. These include glucose with the glucose oxidase method, albumin with the bromcresol green method, and alkaline phosphatase with the sodium thymolphthalein monophosphate method. We have also modified the composition of the reagents used in the determination of protein, inorganic phosphate, and aspartate aminotransferase. A general approach that is useful for adapting a new method to a multichannel AutoAnalyzer system is discussed.

scholarly journals Comparison of three different media for freezing of epididymal sperm from the African buffalo (Syncerus caffer) and influence of equilibration time on the post-thaw sperm quality

2004 ◽  
Vol 71 (3) ◽  
Author(s):  
F.C. Herold ◽  
K. De Haas ◽  
D. Cooper ◽  
B. Colenbrander ◽  
J.O. Nothling ◽  
...  
Keyword(s):  
South Africa ◽  
Sperm Quality ◽  
The Other ◽  
Semen Parameters ◽  
Equilibration Time ◽  
African Buffalo ◽  
Epididymal Sperm ◽  
Progressive Motility ◽  
Reproductive Techniques ◽  
Buffalo Semen

Assisted reproductive techniques might prove themselves useful tools in producing buffaloes free of specific diseases (BFSD), which are in demand in South Africa. Freezing protocols for African buffalo semen must not only result in good post-thaw qualities, but must also be practical. Epididymal sperm from six mature African buffalo bulls was collected, diluted with three different semen extenders and frozen. Pre-freezing equilibration times between 2 and 9 h were tested. Total and progressive motility, longevity and acrosomal integrity were measured and compared. The use of TriladylTM proved to result in better post-thaw parameters than the other two diluents. Equilibration times of between 4 and 9 h did not influence post-thaw sperm qualities significantly. For some of the treatments, exposure to semen extenders before freezing for less than 4 h resulted in inferior post-thaw semen parameters.

scholarly journals The use of insulin-like growth factor 1 improved the parameters of the seminogram in a patient with severe oligoasthenoteratozoospermia

2019 ◽  
Vol 7 ◽  
pp. 2050313X1983415
Author(s):  
José Arturo Mora Rodríguez ◽  
Leonardo M Porchia ◽  
Felipe Camargo ◽  
Esther López-Bayghen
Keyword(s):  
Growth Factor ◽  
Assisted Reproductive Technologies ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Reproductive Technologies ◽  
Quality Parameters ◽  
Intradermal Injection ◽  
Sperm Parameters ◽  
Insulin Like Growth Factor ◽  
Progressive Motility

Male patients suffering from oligoasthenoteratozoospermia typically failed to achieve pregnancy, even with assisted reproductive technologies. Growth hormone and insulin-like growth factor 1 have been shown to regulate sperm quality parameters; therefore, the insulin-like growth factor 1 supplement could improve sperm parameters. Here, we determine the effect insulin-like growth factor 1 has on sperm parameters in a patient suffering from oligoasthenoteratozoospermia. A 47-year-old male was administered once a day 1.5 IU of insulin-like growth factor 1 by intradermal injection for 2 months. Seminogram analysis was performed before and after. Treatment with insulin-like growth factor 1 resulted in a 15.5-fold improvement in sperm concentration (1.1 × 106 vs 18.3 × 106 per mL), 71.4% change in volume (0.7 vs 1.2 mL), increased progressive motility (2% vs 43%), and the total volume of sperm with progressive motility (0% vs 23.6%). Here, we show that administering a daily dose of insulin-like growth factor 1 can improve sperm quality parameters.

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