Semen Dilution Research Articles

THE WORK OF RESEARCHS AT THE DEPARTMENT OF ANIMAL REPRODUCTION BIOTECHNOLOGY OF THE NAAN INSTITUTE OF ANIMAL BREEDING IN DEVELOPING THE SPHERE OF PIG REVIVAL

The scientific contribution of Researchs of the laboratory of artificial fecundation of farm animals of the Institute of Animal Sciences NAAS (now the Department of reproduction biotechnology of farm animals) in the development of artificial fecundation was analyzed as a method of large-scale breeding in the national livestock breeding. The work of Researchs on the development of industry fundamental and applied research on the study of the physiology of pig reproduction was presented. The development of the Laboratory of Boar Reproduction has been examined from the moment of its existence up to the present time. The scientific research of the laboratory Researchs during the postwar years were covered aiming at the integration of the method of artificial fecundation into the farms of the country. Research T.M. Kozenko suggested glucose-citrate-yolk medium for dilution and storage of boar semen (1950). The treasury of his inventions were included wooden stuffed pigs, an artificial vagina for obtaining semen from boars, a glass device for sow fecundation, etc. The contribution of Professor I. V. Smirnov to the development of biotechnological science has been evaluated. His name related to the development of the method of long-term semen storage, improvement of techniques and technology of artificial fecundation, the development of the bases of reproductive biology of farm animals. The researches made by the Research were the catalyst for developing the methods of freezing and long-term storage of sperm of begetters by the Researchs of the Laboratory of Biology of Reproduction and Artificial Fecundation of Farm Animals Serdyuk S. I. and Belikov A. A. (1972). The priority developments of the employees of the Laboratory of Boar Reproduction headed by Prof. S. I. Serdiuk, Doctor of Veterinary Sciences, were outlined. They carried out complex researches concerning reproduction and artificial fecundation of pigs, namely quality estimation of boar semen using optical standard, deep freezing of boar semen in liquid nitrogen at -196ºС. There were developed and integrated into practice devices for artificial fecundation of pigs POS-5, system of artificial fecundation of pigs on large specialized complexes. The Researchs of the laboratory have enriched the national science with new progressive developments, introduced progressive methods of reproduction in breeding farms of Ukraine.

The effects of egg yolk-based and egg yolk-free diluents on the post-thaw quality of bull spermatozoa

The aim of this study was to evaluate the effects of 3 different commercially available extenders – AndroMed® (soy lecithin-based), Optidyl® (with the addition of ionized egg yolk), and BULLXcell® (with the addition of fresh egg yolk) – on bull spermatozoa quality, which was evaluated using the parameters of spermatozoa motility, viability, plasma membrane damage, acrosome damage, and mitochondrial activity after thawing and during a 2 h long thermoresistance test. The spermatozoa quality indicators were appraised by computer-assisted semen analysis and a flow cytometer. Significant differences (P< 0.01) between bulls were registered in all indicators measured. The highest average values of spermatozoa total motility and viability were achieved using BULLXcell® extender (44.33%; 52.06%). Variances in comparing this extender with Optidyl® and AndroMed® were −0.83%, −2.64%; −8.33%, −9.51%. The differences found between the egg yolk-based diluents (BULLXcell® and Optidyl®) and AndroMed® were significant (P< 0.01). Therefore, the more valuable extenders for bull semen dilution were egg yolk-based extenders, which provided higher post-thaw spermatozoa survival and quality than the soy lecithin-based extender.

Evaluation of the effects of hesperidin on fresh and frozen-thawed semen quality using two different cryopreservation methods in Simmental bull.

In the industry of bull semen freezing centers, one-step and two-step semen dilution protocols are two standard and well-known methods in semen freezing process. As the freezing/thawing processes cause detrimental effects on sperm function, the addition of antioxidants can improve sperm characteristics. Hesperidin (Hesp) is an antioxidant used as the male reproductive protective agent. Therefore, the aim of this study was to investigate two different dilution methods, as well as to evaluate Hesp supplementation influence on sperm characteristics in fresh and frozen thawed semen. Semen samples were collected from 12 Simmental bulls. Two separate examinations were conducted in, with and without Hesp supplementation groups. Statistical analysis was performed by an independent t-test, Mann Whitny test, MANOVA and ANOVA tests. In comparison to the one and two-step dilution protocols without Hesp supplementation, the two-step dilution showed greater cryoprotective potential. In the Hesp supplemented group, each semen sample was divided into six equal parts for experimental groups (dilution step method/µM of Hesp). In the both one and two step dilution protocols, significant improvements were detected in semen motility parameters by Hesp administration. Also, oxidative stress status was reduced in seminal plasma of Hesp treatment groups. Interestingly, in comparison with Hesp dosage, 1µM was shown to have greater semen cryoprotective potential. In conclusion,

닭 신선 정자 운동성과 첨체안정성에 보존 온도, 희석제 및 용기가 미치는 영향

Chicken spermatozoa have the ability to survive in low-temperature environments; however, the effects of low temperature on sperm motility and acrosome damage have not been studied in detail. The present study investigated semen longevity following dilution of rooster semen with Beltsville Poultry Semen Extender (BPSE) and Lake extender in preservation vessels (1.5 mL e-tube and 0.5 mL straw). Spermatozoa motility in the closed-type vessel (0.5 mL straw) was higher than that in the 1.5 mL e-tube on day 3 of preservation (68.6±3.1% vs. 22.1±5.7%). The motility of rooster semen diluted with BPSE in 0.5 mL straw was also higher than that of the Lake extender on day 3 of preservation (57.7±5.6% vs. 37.7±5.4%). Furthermore, acrosome intactness was higher in 0.5 mL straw than in the 1.5 mL e-tube, and the rate of acrosome cap damage increased with preservation days. The present study demonstrates that a closed 0.5-mL straw vessel could be used for low-temperature semen preservation, with an increased motility rate and acrosome integrity in fresh rooster semen.

Optimization of Turkey Semen Dilution Rate and Liquid Storage Period At 4-8oc With Plant Based Extender For Optimum Fertility

Optimization of Turkey Semen Dilution Rate and Liquid Storage Period At 4-8oc With Plant Based Extender For Optimum Fertility

Effect of Different Pre-Freezing Time on Quality of Frozen Fat-Tailed Ram Semen

This study aimed to determining the effect of temperature changes on the sperm quality of fat-tailed sheep during the freezing process using a microcontroller. This study was conducted from April to October 2018 at the Laboratory of Animal Physiology and Reproduction, Faculty of Animal Science UGM. The study was used twelve mature ram. The methods were conducted by collecting semen using artificial vagina, semen dilution, freezing semen, thawing and semen quality test. The pre-freezing temperature time were grouped for 6, 9, and 12 minutes. Data of frozen semen quality (motility, viability, and abnormality) the data were analyzed using one-way ANOVA and the study was arranged using Completely Randomized Design (CRD). The average of spermatozoa motility after semen freezing with a 5 to -140°C decrease in pre freezing temperature for 12 minutes (50±5.3%) was significantly different (P<0.05) compared to 9 minutes (48±4.8%) and 6 minutes (43±4.8%). The average of spermatozoa viability after semen freezing with a decrease in pre freezing temperature of 5 to -140°C for 12 minutes (55±4.7%) was significantly different (P<0.05) compared to that for 9 minutes (52±3.5%) and 6 minutes (49±5.7%). The average of spermatozoa abnormality after freezing with a decrease in pre freezing temperature of 5 to -140°C for 6, 9, and 12 minutes was not significantly different (10±2.4%, 9±0.8%, and 10±0.9%, respectively). Based on the findings, it is possible to conclude that semen freezing at a lower pre-freezing temperature of 0 to -140°C for 12 minutes can improve the quality of freezing results.

The Effect of Long Shelf Life of Duroc Pig Male Semen Diluted Using Tris-Egg Yolk-Young Coconut Water on The Value of Viability, Abnormality and pH

This study aims to determine the effect of shelf life of duroc boar semen which was diluted using tris-egg yolk-coconut water thinner on the value of viability, abnormality, and acidity (pH). The semen used is in the form of fresh semen from 2-year-old duroc pigs. Semen was collected by a manual method using artificial broodstock. This research was conducted in November 2020 using an experimental method according to the Completely Randomized Design (CRD) procedure with 4 treatments and 4 replications so that there were 16 experimental units. Each treatment is P0: storage of semen for 0 hours, P1: storage of semen for 24 hours, P2: storage of semen for 36 hours, P3: storage of semen for 48 hours. The results showed that the use of tris-egg yolk-coconut water diluent had a significant effect (P <0.05) on viability, abnormality, and degree of acidity (pH). The mean value of spermatozoa viability was P0: 96%, P1: 93.75%, P2: 84.5%, and treatment P3: 79.5%. The mean value of spermatozoa abnormality in treatment P0: 5.25%, P1: 6.5%, P2: 11.25%, and treatment P3: 14.75%. The average value of the degree of acidity (pH) of semen P0: 8.3 P1: 7.2; P2 7,3 and P3 treatment: 7,25. It can be concluded that the dilution of semen using tris-egg yolk and coconut water can maintain the value of viability, abnormalities of spermatozoa, and degree of acidity (pH).

Molecular Identification and Antibacterial Activity Analysis of Blue Fox (Vulpes lagopus) β-Defensins 108 and 122.

Simple SummaryThe blue fox (Vulpes lagopus) is an important fur animal in China; its reproductive performance is directly related to the economic benefits of breeding. β-Defensins can protect the male reproductive system from bacterial invasion, maintain the stability of reproductive tract microenvironment and improve semen quality in mammals. Few studies have proposed to investigate the antibacterial effect of Vulpes lagopus beta-defensin (vBDs). In this study, we analyzed the antibacterial activity of recombinant vBD108 and vBD122 protein in vitro by an antibacterial activity analysis experiment. Our preliminary results demonstrate that the two vBDs have good antibacterial activity. The blue fox β-defensins may be used as an extender component of the semen diluent to protect semen from bacterial infection.The blue fox (Vulpes lagopus), a fur-bearing animal, is an important component of the breeding industry in China. Semen quality is a key factor for the reproductive process and the breeding effectiveness of the farmed blue fox. However, bacterial contamination in semen samples utilized in artificial fertilization is very common. The β-defensins, a class of important antimicrobial peptides in mammals, could protect the reproductive system of male animals from bacterial invasion, maintain the stability of the genital tract microenvironment and improve semen quality. In this study, molecular cloning and bioinformatics analysis were conducted to analyze the protein structure and function of blue fox β-defensin 108 (Vulpes lagopus beta-defensin 108, vBD108) and 122 (Vulpes lagopus beta-defensin 122, vBD122). To evaluate the bacteriostatic effect of recombinant vBDs (Vulpes lagopus beta-defensins) protein, varying concentrations (0, 25, 50, 100, 200 µg/mL) were taken to evaluate the effects on Escherichia coli and Staphylococcus aureus at different times (0, 2, 4, 6, 8, 12 h). The results showed that vBD108 and vBD122 existed in different forms in protein structure and had antibacterial activity. Both proteins, at 50 µg/mL, had efficacious bacteriostatic activity. This study shows that recombinant vBD108 and vBD122 proteins have good antibacterial activity in vitro. This implies a potential role in improving semen quality and hygienic measures in the process of artificial insemination as an extender of semen dilution with antibacterial activity.

Towards standardization of the cryopreservation procedure of cultured pikeperch (Sander lucioperca) semen

Within the study, attempts towards standardized cryopreservation procedure for cultured pikeperch semen were undertaken. The specific aims of this study were to examine the effects of final glucose and sperm concentrations on the cryopreservation success of cultured pikeperch semen measured by sperm quality indices. We also examined the effect of different solutions for activation of sperm motility as well as tested the effect of the dilution of semen in Kurokura solution on the cryopreservation success of pikeperch. The final glucose concentrations within the range 0.11–0.15 M in 7.5% methanol achieved good results for the percentage of sperm motility after freezing/thawing, with the highest values at 0.13 M (44 ± 12%). The highest sperm motility after freezing/thawing (56–58%) was observed within the range of 3.0 to 4.0 × 109 spermatozoa ml−1. Dilution of semen in seminal plasma resulted in activation of spontaneous sperm motility. Moreover, the extender used for cryopreservation caused activation of sperm motility at a level similar to that of an activation solution. Cryopreserved sperm was characterized by sperm motility similar to that of equilibrated semen. A significant decrease in the sperm motility was recorded in samples not diluted in immobilizing solution after 10 min of post-thaw storage compared to values recorded immediately after thawing. These differences were not observed in semen samples diluted in Kurokura solution. In our opinion, development of standardized cryopreservation protocol presented in this study is a prerequisite for the future implementation of cryopreserved semen in hatchery practice. However, control of spontaneous motility seems to be an important challenge for further improvement of cryopreservation techniques for pikeperch semen.

The influence of semen collection frequency on deleterious the interaction between the enzymes of the bulbourethral gland and egg yolk during the dilution of buck semen

The study aimed to evaluate the influence of semen collection frequency at different protocols on deleterious the interaction between the enzymes of the bulbourethral gland and egg yolk during the dilution and of buck semen. Five bucks aged 2.5-3 years were included in this study. Four different protocols of semen collection frequency were applied; once a week, twice a week at an interval of two days, thrice a week at an interval of one day, and twice daily at an interval of one hour. Buck semen ejaculates were evaluated using computer assisted sperm analysis that includes the percentage of motile sperm (MS), percentage of progressive motile sperm (PMS), velocity curvilinear (VCL), velocity average path (VAP), velocity straight line (VSL), linearity (LIN), straightness (STR), wobble (WOB), amplitude of lateral head movement (ALH), and beat cross frequency (BCF). Results showed that sperm motility parameters had decreased significantly after dilution. Sperm motility parameters (VCL, VSL, VAP and ALH) of the second ejaculate were higher significantly (P<0.05) after dilution, when ejaculates collected twice and thrice a week and twice a day. Conversely, motility parameters of third ejaculates were less than those of first and second ejaculates when ejaculates collected thrice a week. In conclusion, there are effects of semen collection frequency on the ability of buck semen dilution, and the best results were when ejaculates had collected twice a day at an interval of one hour, accordingly we recommended this protocol for preparation of buck semen for artificial insemination.

СРАВНИТЕЛЬНАЯ ХАРАКТЕРИСТИКА СПЕРМЫ БАРАНА, ЗАМОРОЖЕННОЙ В РАЗНЫХ ЭКСТЕНДЕРАХ

Numerous studies have proved the necessity of egg yolk in synthetic media to dilute semen before its cryopreservation. However, at the same time, it has been demonstrated that its use can adversely affect the quality of frozen-thawed sperm. The present study compares the main quality parameters of ram sperm frozen using TRIS-based diluent with egg yolk and two egg yolk-free diluents (OvixCell® and AndroMed®). A slower deterioration in the kinematic performance of sperm cryopreserved in TRIS diluent with native egg yolk confirmed higher cryoprotective performance compared to commercial extenders containing no egg yolk. Significantly higher total and progressive motility was observed in TRIS-based medium with egg yolk (P&lt; 0.05). This advantage was maintained after four hours of incubation and became more significant at the end of cultivation (after six hours) (P&lt; 0.01). Thus, ram sperm frozen in egg yolk medium retained better motility than in egg yolk-free extenders, which allows predicting its higher bioavailability. Assessment of some semen parameters using CASA showed that there were no significant differences in motility between the three extenders immediately after thawing the straws. When assessed two hours after thawing, a diluent containing egg yolk (TRIS-based) was found to have higher results for some of the examined traits than phospholipid diluents.

Efficiency of Different Diluents and Dilution Rates on the Fertilization Potential of Chicken Spermatozoa

Background: Artificial Insemination (AI) has been well established to obtain better fertility than natural mating. Semen extension and preservation play a major role in AI technique. The concentrated nature of chicken semen entails dilution of semen for easy handling and storage. With the availability of a wide range of extenders, exploring various extenders and dilution rates supplement the reproductive efficiency achieved through modern AI techniques.Methods: Pooled semen samples obtained from twenty White Leghorn breeder males were split equally into three portions, each for CARI poultry semen diluent, EK extender and Tselutin extender. At 1:2 dilution rate, the samples were subjected to sperm motility analysis from 0 to 96 h and AI was targeted at 0 and 24 h of semen collection. Fertility was assessed by candling of eggs collected from 2 to 8 days post insemination at 9th day of incubation. Further investigation of different dilution rates (1:2, 1:4, 1:6, 1:8, 1:10, 1:12, 1:14, 1:16, 1:18, 1:20) on fertility of fresh semen was done using the superior diluent.Result: Upon analysis, sperm motility showed no significant difference among the diluents at 0 h storage while CARI poultry semen diluent showed superior motility (%) at the subsequent storage periods, followed by EK and Tselutin extender. Higher fertility (p less than 0.05) was expressed in the diluents of CARI and EK at 0 h storage, whereas CARI poultry semen diluent showed superior fertility pattern (p less than 0.05) followed by EK and least by Tselutin extender at 24 h semen storage. Further investigation of different dilution rates using CARI diluent, exhibited fertility (p less than 0.05) of 90% and higher at 1:2 and 1:4 dilutions, followed by 1:6 and 1:8 at 2 to 6 days of fertile period.

Jambul (Syzygium cumini) Pulp Extract Enhances Viability, Motility, andIn VitroFertilizability of Cryopreserved Bovine Semen

To study the ameliorative capacity of Syzygium cumini fruit Pulp Extract (SPE) on cryopreserved bovine semen, three ejaculates each of three fertile bulls (total nine samples) were collected. Each sample was mixed with semen dilution medium in 1:2 ratios, centrifuged, and 50% of the supernatant was discarded. The remaining sample was mixed with semen cryopreservation extender (SCE) in 1:4 ratios. Three aliquots (0.3 mL) of each extended semen sample were further diluted with (1) 0.7 mL SCE (SCE group), (2) 0.7 mL SCE containing SPE equivalent to 7 ppm total antioxidants (SPE7 group), and (3) 0.7 mL SCE equivalent to 14 ppm antioxidants (SPE14 group). For each aliquot, semen quality was analyzed in situ as well as after cryopreservation. Results show significant improvement in sperm membrane integrity, motility, and fertilizability in SPE7 (63.5% ± 2.04%, 56.22% ± 1.7%, 51.1%, respectively) and SPE14 (68% ± 1.12%, 63.67% ± 1.06%, 57.7%, respectively) than the SCE group (62.33% ± 1.22%, 54.9% ± 1.34%, 48.8%, respectively). Similarly, significant improvement in the mean percent number of progressively motile sperm at the expense of the mean percent twitching, shaking, and whirling spermatozoa, both before and after cryopreservation, were observed in SPE7 (72 ± 1.06, 61.2 ± 1.2) and SPE14 (78.89 ± 1.12, 67 ± 1.32) groups to that of the SCE (59.6 ± 0.97, 52.3 ± 3.5). In addition, the mean percent of progressively motile spermatozoa showing 20 μm/s or above velocity, before and after cryopreservation, were significantly higher in SPE7 (58.9 ± 4.7, 43.6 ± 1.5) and SPE14 (66.9 ± 0.8, 51.3 ± 0.8) groups than that of the SCE (42.3 ± 3.1, 30 ± 2.4). These findings indicate that SPE can significantly improve various quality parameters of the cryopreserved bovine semen.

Total Bacteria of Post-Thawing Boer Buck Semen with Addition of Sweet Orange Essential Oil to Tris Yolk Extender

Aims: To determine the effect of the addition of sweet orange essential oil to the extender (tris yolk extender without the addition of antibiotics/antibacterial) to total bacteria Boer Buck frozen semen.&#x0D; Study Design: Randomized Block Design.&#x0D; Place and Duration of Study: Sample: Laboratory Reproduction of Loka Penelitian Kambing Potong Sei Putih Indonesia, between January and March 2019.&#x0D; Methodology: The research procedure starts with the preparation of semen extender, collection of fresh semen, dilution of semen, equilibration, freezing of semen, and thawing. This research obtained conducted using a Randomized Block Design consisting of 5 treatment levels and five replications. Semen storage using 3 Boer buck, which done for three days. As a treatment is the addition of sweet orange essential oil as much as (P0) 0%, (P1) 0,25%, (P2) 0,5%, (P3) 0,75% and (P4) 1% on the tris yolk extender. The observed variables was total bacteria evaluated before freezing and after freezing (Post-Thawing).&#x0D; Results: The results showed that the addition of sweet orange essential oil had a very significant effect (P &lt;0.01). The results of adding sweet orange essential oil to the extender (Post-Thawing) were 78.6 x 102 CFU (P0), 76 x 102 CFU (P1), 73,2 x 102 CFU (P2), 71 x 102 CFU (P3) and 68.6 x 102 CFU (P4).&#x0D; Conclusion: The best values the total bacteria 68.6 x 102 CFU (P4). It can conclude that the best result of adding sweet orange essential oil to tris yolk extender is (P4) 1%.

Standardization of quail egg-yolk concentration with tris buffer for avian semen dilution and preservation for optimum fertility

Standardization of quail egg-yolk concentration with tris buffer for avian semen dilution and preservation for optimum fertility

Effect of semen extenders and storage time on quality of Muscovy duck (Cairina moschata) drake semen during the entire reproductive season.

Avian semen dilution with appropriate extender allows to prolong the fertilizing ability of sperm stored in vitro. In the present study, the impact of extenders and time of storage on morphology of Muscovy duck (Cairina moschata) drake semen were examined. Semen was collected twice a week, using male stimulation by a female method, from 12 adults (29weeks old) drakes kept individually in cages, under controlled environmental conditions. Freshly collected, pooled ejaculates were divided into three part: neat undiluted sample, and diluted 1:1 with Schramm (SCH) or Watanabe (W) extender and stored at 4°C. Morphological examination of all samples was conducted after dilution and then, after 3 and 6hr of storage. The storage of undiluted semen caused decrease (p≤.01) in live morphologically normal sperm, from 79.73% in the freshly collected ejaculates to 55.75% and to 12.12% after 3 and 6hr of storage, respectively (average calculated for the entire reproductive season). In the semen diluted with Schramm's extender the adequate values attained 86.84, 79.65 and 61.66%, and using Watanabe extender 84.77, 83.58 and 75.25%, respectively. The period of semen storage and the type of extender caused significant (p≤0,05; p≤0,01) changes in sperm morphology. The longer period of storage contributed to the decrease in number of morphologically normal sperm, whereas their content in Watanabe extender after 3 and 6hr of storage was higher (p≤.01) than in semen diluted in Schramm extender.

Comparison between Tris-buffer and INRA-82 extenders on the quality of chilled rabbit spermatozoa

&lt;p&gt;The aim of the present study was to compare the effects of tris-buffer and INRA-82 extenders on the quality of cooled rabbit spermatozoa. Pooled semen samples were collected from eight New Zealand White rabbit bucks, divided into three groups and diluted 1:5 with three different semen extenders: INRA-82, tris-citrate glucose (TCG) and tris-citrate trehalose (TCT). Following dilution, the samples were stored at 5°C for up to 48 h. Sperm motility was recorded at 24, 36 and 48 h post-cooling. Furthermore, sperm viability, morphology, membrane integrity, acrosome status and DNA integrity were assessed at 24 h post-cooling. We found that progressive motility percentages were significantly higher (&lt;em&gt;P&lt;/em&gt;&amp;lt;0.05) in samples diluted in INRA-82 (61.00, 53.50, 44.00% at 24, 36 and 48 h post-cooling, respectively) than those diluted in either TCG (39.25, 32.00, 19.75%) or in TCT (47.25, 40.50, 29.00%). We also reported that sperm viability, percentage of normal spermatozoa, percentage of spermatozoa with intact acrosome and DNA integrity after 24 h cooling were significantly higher (&lt;em&gt;P&lt;/em&gt;&amp;lt;0.05) in INRA-82 diluted samples compared to TCG and TCT diluted ones. In summary, our results show that dilution of rabbit semen in INRA-82 improved sperm physiological parameters post-cooling compared to those diluted in TCG or TCT. Our findings also suggest that INRA-82 is a promising diluent that can be used effectively to maintain the viability of chilled rabbit semen.&lt;/p&gt;

Effect of semen extenders on viability of ISA Brown and Hubbard Flex roosters' sperm stored for 24 h

Artificial insemination is used in almost 95% of turkey reproductive flocks and is becoming more important in chickens, particularly broiler breeders, as well as in assisted reproduction of wild birds kept in breeding centers. Diluted semen is recommended for artificial insemination. Pooled semen samples collected twice a week by dorso-abdominal massage from 2 chicken lines: laying—ISA Brown (ISA-B) and meat type—Hubbard Flex (H-F) were divided into 5 parts: neat semen and diluted in 1:2 ratio with 4 extenders: basic EK; EK + 1 μg/mL organic selenium and 8 μg/mL vitamin E; EK + 10 mg/mL of royal jelly; and EK + 0.25 g/mL of lyophilized bovine colostrum. Diluted semen samples were evaluated after 15 min and then 24 h storage at 4°C. Sperm concentration, motility, motility parameters (with Sperm Class Analyzer), and morphology were evaluated in the neat semen, whereas in diluted and stored samples, the last 3 traits were determined. In case of both lines, dilution did not affect (P > 0.05) the number of live normal cells (78.0–81.1% in ISA Brown and 73.8–68.7% in Hubbard Flex) in relation to neat semen; however, bovine colostrum addition increased (P < 0.05) the percentage of bulb head sperm (5.7 vs. 10.0% and 12.1 vs. 17.6%, for ISA and Hubbard, respectively) and decreased sperm motility (67.4 vs. 92.9% and 67.3 vs. 98.5% for ISA and Hubbard). The 24 h storage of neat semen and semen diluted with colostrum caused (P < 0.05) the unfavorable changes in all evaluated traits and both chicken lines, whereas semen dilution with remaining extenders decreased the percentage of live normal cells (by 18.8–23.4% ISA and by 20.9–25.5% Hubbard) but did not affect sperm motility (81.5–87.6% for ISA and 81.1–96.6% for Hubbard). Sperm motility and motility parameters depended both on the extender and chicken line.

Antimicrobial Resistance as a Problem for the Quality of Boar Semen

Abstract The aim of the study was to determine whether the bacteria from the environment and from the mucous membrane of the boar prepuce have antimicrobial resistance and whether the result obtained is similar/same to the bacteria that can be found in native boar semen. The study addresses the problem of the presence of primarily resistant bacterial strains in the boar sperm, which, due to their reduced sensitivity, cannot be suppressed by antibiotics used in the semen dilution agent, as well as to emphasize the importance of microbiological monitoring of the boar mucous membranes and ambient surfaces before and during their exploitation. Such an examination could contribute to the interchangeable design of the dilution agent for the boar semen relative to the antibiotic content.Resistant strains of bacteria from prepuce swabs and swabs taken from the facility, as well as from native boar semen were isolated. The presence of these bacteria affected the quality of the semen. In conclusion, it should be pointed out that bacterial monitoring of the prepuce and surface of the facility can indicate possible problems related to the quality of semen, and that the design of the dilution agent for boar semen should be adjusted to the established resistance of isolated bacteria.

Comparison of Crime-lite® 82S, Polilight® PL400 and Polilight® PL500 for the detection of semen and saliva stains

ABSTRACT Alternative light sources (ALS) are widely employed in forensic investigations for the identification of biological fluids, such as saliva and semen. While metal halide arc lamps are a common light source for many ALS, high intensity light-emitting diode (LED) globes are a more energy efficient alternative. To date, ALS operating with different light sources have not been evaluated against each other, and this study is the first to complete a side-by-side comparison of the arc lamps, Polilight® PL400 (PL400), Polilight® PL500 (PL500) and the LED-based Crime-lite® 82S (blue) (CL82S) for the detection of semen and saliva dilutions on various fabric materials. The absence or presence of fluorescence were scored independently by four experienced item examination scientists to reduce bias. The number of fluorescence stains detected using the CL82S (244 observations) was higher and brighter in intensity compared to those visualized using PL400 and PL500 (183 and 202 observations, respectively). Notably, dilutions of saliva which were not visible under PL400 and PL500 on a range of fabrics were detected under CL82S, which was statistically significant (p < 0.003). Taken together, CL82S is a better screening tool than the PL400 and PL500 for detection of dilute semen and saliva stains on various fabric types.