Authors:
Malam Abulbashar Mujitaba Doctoral School of Animal Science, University of Debrecen, Böszörményi Street 138, H-4032, Debrecen, Hungary
Institute of Animal Science, Biotechnology and Nature Conservation, Faculty of Agricultural and Food Sciences and Environmental Management, University of Debrecen, Böszörményi Street 138, H-4032 Debrecen, Hungary

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Gabriella Kútvölgyi Department of Precision Livestock Farming and Animal Biotechnics, Institute of Animal Sciences, Hungarian University of Agriculture and Life Sciences, Kaposvár Campus, H-7400 Kaposvár, Guba S. U. 40, Hungary

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https://orcid.org/0009-0001-1591-998X
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Viktória Johanna Debnár Department of Precision Livestock Farming and Animal Biotechnics, Institute of Animal Sciences, Hungarian University of Agriculture and Life Sciences, Kaposvár Campus, H-7400 Kaposvár, Guba S. U. 40, Hungary

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Alexandra Tokár Hungarian University of Agriculture and Life Sciences, Festetics György Doctoral School Deák Ferenc utca 16., 8360, Keszthely, Hungary

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János Posta Institute of Animal Science, Biotechnology and Nature Conservation, Faculty of Agricultural and Food Sciences and Environmental Management, University of Debrecen, Böszörményi Street 138, H-4032 Debrecen, Hungary

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Szilárd Bodó Department of Precision Livestock Farming and Animal Biotechnics, Institute of Animal Sciences, Hungarian University of Agriculture and Life Sciences, Kaposvár Campus, H-7400 Kaposvár, Guba S. U. 40, Hungary

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Nóra Vass Institute of Animal Science, Biotechnology and Nature Conservation, Faculty of Agricultural and Food Sciences and Environmental Management, University of Debrecen, Böszörményi Street 138, H-4032 Debrecen, Hungary

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Abstract

This study was conducted to develop ideal post-mortem gamete retrieval and conservation methods to establish a Hungarian ex-situ in vitro gene bank. Pairs of testes from German Mutton Merino (n = 7) and Hungarian Black Racka (n = 7) rams were collected at a slaughterhouse, transported to the laboratory and stored overnight (4−5 °C) before processing. Post mortem ram epididymal spermatozoa (REPS) were obtained from the cauda epididymidis by slice or incision methods. Fresh samples were extended to 200 × 106/mL cell concentration, filled into mini straws and equilibrated at 5 °C for 2 h. Freezing was performed manually in a Styrofoam box. The fresh and post-thaw total motility, progressive motility and kinematic parameters of REPS were assessed using the CASA technique. The collection method did not affect significantly the fresh and post-thaw motility and kinematic parameters. Merino had higher (P < 0.05) testicular weight. Racka had significantly better fresh and post-thaw linear movement but had statistically the same (P > 0.05) cryotolerance as Merino. In conclusion, both collection methods were found suitable for REPS retrieval. The REPS from Racka exhibited better linear movement values than those from the Merino breed. The cryotolerance of REPS of both breeds was comparable.

Abstract

This study was conducted to develop ideal post-mortem gamete retrieval and conservation methods to establish a Hungarian ex-situ in vitro gene bank. Pairs of testes from German Mutton Merino (n = 7) and Hungarian Black Racka (n = 7) rams were collected at a slaughterhouse, transported to the laboratory and stored overnight (4−5 °C) before processing. Post mortem ram epididymal spermatozoa (REPS) were obtained from the cauda epididymidis by slice or incision methods. Fresh samples were extended to 200 × 106/mL cell concentration, filled into mini straws and equilibrated at 5 °C for 2 h. Freezing was performed manually in a Styrofoam box. The fresh and post-thaw total motility, progressive motility and kinematic parameters of REPS were assessed using the CASA technique. The collection method did not affect significantly the fresh and post-thaw motility and kinematic parameters. Merino had higher (P < 0.05) testicular weight. Racka had significantly better fresh and post-thaw linear movement but had statistically the same (P > 0.05) cryotolerance as Merino. In conclusion, both collection methods were found suitable for REPS retrieval. The REPS from Racka exhibited better linear movement values than those from the Merino breed. The cryotolerance of REPS of both breeds was comparable.

Introduction

Autochthonous sheep breeds are interesting because they are thrifty and hardier than commercial or modern sheep breeds. Unfortunately, most of them are rarely bred nowadays (https://dengarden.com/agriculture/Rare-and-Endangered-Sheep-Breeds). The Hungarian Racka, also called Hortobágyi Racka, is an important native sheep breed reared by farmers for meat, milk and wool production. It is a hardy, medium-sized sheep (males and females weigh average 60 and 40 kg, respectively), with heights of 61–76 cm at withers and having spiral, V-shaped horns in both sexes (60 and 30 cm for rams and ewes) ensuring an amazing appearance. Racka sheep are mostly of two distinct coat colours, solid black or white, with distinctive medium-fine spiral wool (Zsolnai et al., 2021; Anon et al., 2023). Racka rams have a seasonal reproductive cycle as indicated by changes in their scrotal circumference, sperm quality and quantity across the year. The highest scrotal circumference was observed in September, and the lowest in January. Total sperm count per ejaculate is highest in autumn and the lowest in spring, while the lowest sperm concentration and the highest percentage of abnormal spermatozoa have been recorded in winter (Sarlós et al., 2013).

German Mutton Merino (called Merinofleischschaf in German) is a large-framed white sheep reared for meat and wool. An old German Mutton Merino ram weighs 120−140 kg, while the ewes are of 70–80 kg (Mason, 1996). The sperm motility is better in the summer and autumn months, while it decreases during winter. In this breed, the sperm concentration is 3,200–4,500 × 106/mL, with the highest values recorded from April to October and the lowest ones between November and March at the climate of Israel (Amir and Volcani, 1965). In Spain, the breeding season is from October to February, when highest motility and progressive motility of the ejaculated spermatozoa have been observed (Arando et al., 2019).

According to the 2020 and 2021 local risk status reports DAD-IS (Domestic Animal Diversity Information System; Retrieved July 21, 2023, from: https://www.fao.org/dad-is/sdg-252/en/), both the population and the male to female ratio of certain Hungarian sheep breeds are declining This leads to the extinction or decline of certain important genotypes including Alföldi suta racka (extinct; population:30/30 vs. 26/26 and male to female ratio 4/26 vs. 0/26); Cikta (endangered; population:826/826 vs 789/789 and male to female ratio 28/794 vs 22/767); and Cigàja (at risk; population:3373/3373 vs. 3191/3191 and male to female ratio 163/3210 vs. 85/3106) for 2020 vs 2021, respectively. In the case of the Hungarian Racka breed, the population declined between 2019 and 2020 (8547/8547 vs. 5898/5898), while the male to female ratio increased in favour of males (270/8277 vs. 298/5600). A cryoconservation program exists for these breeds (DAD-IS, 2020). Similarly, in situ gene conservation programs of the breeds run in different national parks in Hungary. The main objective is to keep purebred animals in the colour variants (black and white). The main selection criteria include lamb weight gain, yearling weight and phenotypic characteristics such as wool length, colour and horn appearance. Both the in situ and ex-situ programs are supported by tenders entitled “In situ preservation of the genetic stock of protected native and endangered agricultural animal breeds” and “Ex situ or in vitro preservation of the genetic stock of protected native and endangered agricultural animal breeds, and support for advisory activities preventing genetic narrowing”. This led to the establishment of an ex situ in vitro gene bank consisting of ejaculated spermatozoa of the Hungarian Racka, Cikta and Cigája breeds in cooperation with the Hungarian Sheep and Goat Breeders' Association and the Research Institute for Animal Breeding, Nutrition and Meat Science of the National Agricultural Research and Innovation Centre, in Herceghalom during 2014–2017.

Epididymal spermatozoa (EPS) are considered the cheapest potential source of male gametes for assisted reproductive technology studies and genome resource banking of accidentally dead, severely injured, endangered or threatened elite males (Goovaerts et al., 2006). They can be retrieved differently depending on the species involved or epididymal size. The most used EPS retrieval methods include a) Slicing/mincing (Kaabi et al., 2003; Parra-Forero et al., 2015), b) Incision (Karja et al., 2010; Alvarez et al., 2012; Ahmed, 2019), c) Retrograde flushing via ductus deferens (resulting in less contamination) (Lorraine Leibfried-Rutledge et al., 1997; Bertol, 2016) and d) Floatation (in case of species with tiny testicles) (Bertol, 2016). Good pregnancy rates have been reported using EPS for artificial insemination in sheep (87.5%, 58.5%, and 55.0%) (Ehling et al., 2006; Rickard et al., 2014; Fernández Abella et al., 2015) and goats (61.2%) (Ocampo et al., 2021). Therefore, they can be used to preserve the genetic resources of elite rams. Our literature review, published recently, has revealed that collection of the epididymal spermatozoa is an ideal alternative for gene conservation because it has a recovery rate comparable to the use spermatozoa obtained by the use of artificial vagina (AV) and serves as the easiest and cheapest means of conserving livestock GnR (Mujitaba et al., 2022). Considering the limited published articles on factors affecting the quality of ram epididymal spermatozoa (REPS), we hypothesised that ram breed and collection methods might significantly affect the motility and kinematic parameters of the fresh sample and cryotolerance of REPS. Therefore, we conducted a pilot study out of season to develop an ideal post-mortem gamete retrieving and conservation method to establish a Hungarian ex-situ in vitro gene bank. The specific objectives of the study included to assess the effects of two REPS recovery methods (slicing vs. incision) and breed (German Mutton Merino vs. Hungarian Racka) on the fresh and post-thaw motility and kinematic parameters, and to compare the cryotolerance of the REPS obtained the above ways.

Materials and methods

Media, reagents, and materials

Andromed® semen extender (one-step, 200 mL, Minitube, Tiefenbach, Germany), 0.25 mL transparent semen straws (Minitube, Tiefenbach, Germany) and PBS tablets (Gibco, Lot:2565974) were used for the experiment. The extender was reconstituted according to the manufacturer's guidelines, filled into sterilized 10 mL centrifuge tubes, and stored at frozen condition until required for use. All other plasticware was purchased from Falcon® (Corning, Inc., USA).

Study location, duration and testicle collection

The study was conducted at the spermatology laboratory of the Department of Precision Livestock Farming and Animal Biotechnics, Institute of Animal Sciences, Kaposvár Campus, Hungarian University of Agriculture and Life Sciences, Herceghalom, Hungary during the non-breeding season. Fourteen pairs of testes from seven 2−5-year-old healthy German Mutton Merino (weighing 80−100 kg) and seven black variants of Hungarian Racka rams (weighing 55−60 kg) were collected at a slaughterhouse in Hungary. The testes, left in the scrotum, were transported to the laboratory in an icepack (4−5 °C) within 2–3 h, stored overnight in a refrigerator (4−5 °C) to simulate field conditions, and processed the following day as described by Egerszegi et al. (2012).

Epididymal sperm collection methods

After removing the scrotal sac and lamina parietalis of tunica vaginalis, the testis with epididymis was weighed using a digital scales. Each epididymis was carefully separated and spermatozoa from both caudae epididymidis (CE) of the same ram were retrieved randomly by either the slicing or incising method.

The slicing method (SL). The visceral layer of tunica vaginalis covering the CE is carefully removed to avoid blood contamination. The stripped CE was washed with PBS, then cut out and sliced with a scalpel in a Petri dish containing 3 mL of Andromed® semen extender. The sliced CE was left in the extender at room temperature for 10 min to enhance spermatozoa collection. It was then rinsed with 2 mL of the semen extender and filtered through gauze sheets (Fig. 1).

Fig. 1.
Fig. 1.

Epididymal sperm collection by slicing (A, B, C, D and J) and incision (E, F, G, H, I and J) methods. A: stripping the cauda epididymis (CE) for slicing; B: the stripped CE ready for slicing; C: slicing the stripped CE; D: rinsing the CE slice; E: the engorged CE ready for incision, F: incision of CE by a deep vertical cut; G: additional small horizontal incisions; H: pressing the incised CE against the bottom of the Petri dish for aiding the release of spermatozoa; I: rinsing the incised CE; H: filtering the spermatozoa

Citation: Acta Veterinaria Hungarica 71, 3-4; 10.1556/004.2023.00945

The incision method (IN). We adopted the procedure published by Ahmed et al. (2019) with little modifications. We used Andromed® semen extender to retrieve spermatozoa and a haemostatic forceps was used to engorge the CE to facilitate emptying the spermatozoa. Subsequently, a single deep longitudinal incision at the ventral part of the CE with fewer blood vessels and 3–4 parallel incisions in the inner part were made using a sterile scalpel. It was then pressed against the bottom of the Petri dish containing 3 mL of Andromed® semen extender to aid in emptying the spermatozoa and then rinsed with 2 mL of the same extender. Finally, a sterile gauze was used to sieve the epididymal tissue debris (Fig. 1).

Epididymal sperm dilution, equilibration, freezing and motility assessment

Sperm concentration was assessed with a Makler counting chamber (Sefi Medical Instruments, Haifa, Israel) using a phase contrast microscope at 200× magnification. Upon establishing the concentration, each sample was extended using Andromed® at room temperature to a final concentration of 200 × 106 spermatozoa/mL and manually filled and sealed using PVA into well-labelled 0.25 mL French mini straws.

The filled and sealed straws were equilibrated in a refrigerator (5 °C for 2 h), and freezing was done manually in a Styrofoam box at 4 cm above liquid nitrogen for 8 min. Finally, the frozen straws were plunged into the liquid nitrogen container for permanent storage. After about 2 weeks, the frozen samples were thawed by warming at 37 °C for 30 s.

The motility and kinetic parameters of the fresh and frozen-thawed spermatozoa were assessed using a CASA AndroVision® system (Minitube, Tiefenbach, Germany). The samples were diluted to a concentration of 50–60 × 106 spermatozoa/mL using the same extender. At least 10 random fields per sample or a total of 500 spermatozoa were analysed for standard motility [total motility (TM, %), progressive motility (PM, %)] and kinematic parameters such as average path velocity (VAP, μm s−1), straight line velocity (VSL μm s−1), curvilinear velocity (VCL, μm s−1), amplitude of lateral head displacement (ALH, μm), beat cross frequency (BCF, Hz), straightness (STR = VSL/VAP × 100, %), linearity (LIN = VSL/VCL × 100, %) and wobble (WOB = VAP/VCL × 100, %) as described previously (Goovaerts et al., 2006; Kang et al., 2018).

Data analysis

Data regarding the fresh and frozen-thawed samples were collected and recorded. The data were tested for normality using Shapiro-Wilk test and were normally distributed. A two-way ANOVA was used to check the effects of the collection method and breed (pooled data of the slicing and incision methods) under the two conditions (fresh and post-thaw) separately at a level of significance set at P < 0.05.

The model for the experiment was Yj = a+bx1+cx2+ x1*x2 where Y = dependant variable (e.g TM), a = intercept, x1 = breed (Merino or Racka) and x2 = method (SL or IN).

The effect of freezing within breed was analysed using a paired Student's t-test as the ejaculate of a ram was separated into two doses. One dose was analysed fresh while the other dose was frozen. In contrast, the cryotolerance of the two breeds was calculated using the following formulae CT = values after thawing/values before freezing × 100% as described by Ehling et al. (2006). The differences between breeds (Merino vs. Racka) were analysed using an independent sample t-test and the significance was checked using a two-tailed test.

Results were presented as means ± standard error (SE). The above-mentioned methods were carried out using IBM® SPSS® statistical software version 29.

Results

Effects of collection method on the fresh and post-thaw ram epididymal sperm standard motility and kinematic parameters

Table 1 illustrates the effects of the collection methods (SL vs IN) on fresh and frozen-thawed REPS. Testicular weight dis not differ between the collection methods (SL vs IN; P > 0.05). Similarly, the collection methods did not have significant (P > 0.05) effects on standard motility and any of the kinematic parameters in case of both the fresh and post-thaw REPS.

Table 1.

Effects of collection methods on testicular weight, standard motility and kinematic parameters of fresh and post-thaw ram epididymal spermatozoa

ParametersFreshP-valuePost-thawP-value
Collection methods (mean ± SE)Collection methods (mean ± SE)
SlicingIncisionSlicingIncision
TW (g)213.45 ± 16.9217.82 ± 16.30.789
TM (%)82.07 ± 2.580.07 ± 2.80.59845.83 ± 5.436.67 ± 4.90.324
PM (%)73.00 ± 2.968.93 ± 3.40.37631.58 ± 5.225.08 ± 4.90.504
VCL (μm s−1)132.61 ± 7.8129.09 ± 9.80.781102.73 ± 5.2106.01 ± 5.90.776
VAP (μm s−1)66.24 ± 3.363.39 ± 4.10.59150.41 ± 2.451.33 ± 2.90.880
VSL (m s−1)47.04 ± 2.344.70 ± 3.20.57238.89 ± 1.939.72 ± 2.70.845
LIN (%)35.57 ± 1.234.29 ± 1.00.36837.42 ± 0.836.67 ± 0.80.587
STR (%)70.86 ± 1.669.43 ± 1.40.48376.50 ± 1.076.42 ± 1.40.913
WOB49.93 ± 0.648.86 ± 1.10.38944.75 ± 3.748.08 ± 0.40.463
BCF (Hz)26.19 ± 0.625.60 ± 0.60.45125.93 ± 0.725.92 ± 0.90.907
ALH (μm)4.86 ± 0.34.7 ± 0.30.6553.85 ± 0.23.83 ± 0.20.811

ALH: amplitude of the lateral head displacement; BCF: beat cross frequency; LIN: linearity, PM: progressive motility, SE: standard error; STR: straightness; TM: total motility, TW: testicular weight, VAP: average path velocity; VCL: curvilinear velocity; VSL: straight line velocity, WOB: wobble.

Testicle numbers: slicing (n = 14), incision (n = 14).

Effects of breed on testicular weight, fresh and post-thaw ram epididymal spermatozoa motility and kinematic parameters

Considering that there was no significant difference (P > 0.05) between the collection methods on any of the parameters, the data were pooled and analysed together to determine breed effects on testicular weight (TW; g), total motility (TM; %) and progressive motility (PM; %) and kinematic parameters of the fresh and frozen-thawed REPS. The results are summarized in Table 2. The TW was found significantly (P < 0.05) higher in the Merino breed than in the Racka. In contrast, REPS from the Racka breed presented several, significantly (P < 0.05) better kinematic parameters (LIN, STR, BCF and ALH) than the Merino breed. All other parameters were not significantly (P > 0.05) different between the breeds.

Table 2.

Effects of breed on testicular weight, standard motility and kinematic parameters of fresh and post-thaw ram epididymal spermatozoa (mean ± SE)

ParametersFreshP-valuesPost-thawP-values
BreedBreed
MerinoRackaMerinoRacka
TW (g)260.50 ± 11.8a170.82 ± 10.1b0.001
TM (%)83.64 ± 2.278.50 ± 2.80.18244.20 ± 6.4739.14 ± 4.480.498
PM (%)73.00 ± 3.168.93 ± 3.30.37630.00 ± 6.9727.14 ± 3.820.702
VCL (μm s−1)140.13 ± 8.5121.58 ± 8.60.152104.76 ± 4.69104.09 ± 5.880.936
VAP (μm s−1)68.39 ± 3.361.24 ± 3.80.18650.05 ± 2.0251.45 ± 2.860.728
VSL (m s−1)46.78 ± 2.344.96 ± 3.20.65937.75 ± 1.6540.41 ± 2.560.456
LIN (%)33.2 ± 1.0b36.6 ± 0.9a0.02235.40 ± 0.7b38.21 ± 07a0.008
STR (%)67.9 ± 1.6b72.43 ± 1.1a0.03274.90 ± 1.377.57 ± 1.10.127
WOB (%)48.43 ± 0.950.36 ± 0.80.12747.30 ± 0.445.79 ± 3.20.698
BCF (Hz)24.86 ± 0.5b26.92 ± 0.6a0.01324.43 ± 0.72b26.99 ± 0.71a0.027
ALH (μm)5.22 ± 0.2b4.35 ± 0.3a0.0224.10 ± 0.163.66 ± 0.140.064

ALH: amplitude of the lateral head displacement; BCF: beat cross frequency; LIN: linearity, PM: progressive motility, SE: standard error of mean; STR: straightness; TM: total motility, TW: testicular weight, VAP: average path velocity; VCL; curvilinear velocity; VSL: straight line velocity, WOB: wobble.

Means in the same row with different superscript within a form (fresh or post-thaw) differ significantly.

Testicle numbers: Merino (n = 14), Racka (n = 14).

The breed had no significant (P > 0.05) effects on the post-thaw standard motility parameters and kinematic parameters except for LIN and BCF, which were significantly (P < 0.01 and P < 0.05, respectively) higher in the Racka breed.

Comparison of the cryotolerance of epididymal spermatozoa from Merino or Racka rams

In Table 3, the comparison of the cryotolerance of REPS from Merino and Racka are presented. Freezing and thawing significantly (P < 0.05) reduced the values of TM, PM, VCL, VAP and ALH but significantly (P > 0.05) increased the STR. WOB and BCF were not affected significantly (P > 0.05) in any of the breeds. The VSL of Merino declined significantly (P < 0.05) following freezing and thawing, while that of Racka did not. The cryotolerance of the REPS from the Merino or Racka breeds did not differ statistically (P > 0.05).

Table 3.

Comparison of the cryotolerance of ram epididymal spermatozoa of Merino and Racka breeds (mean ± SE)

ParametersMerinoP-valuesRackaP-valuesCryotolerance
FreshPost-thawFreshPost-thawMerino CT (%)Racka CT (%)P-values
TM (%)83.64 ± 2.2a44.20 ± 6.5b0.000178.50 ± 2.8a39.14 ± 4.5b0.000152.30 ± 6.949.91 ± 5.30.783
PM (%)73.00 ± 3.1a30.00 ± 6.9b0.000168.93 ± 3.3a27.14 ± 3.8b0.000139.42 ± 8.739.67 ± 5.30.980
VCL (μm s−1)140.13 ± 8.5a104.76 ± 4.7b0.003121.58 ± 8.6a104.09 ± 5.9b0.02076.66.±5.288.44 ± 5.20.133
VAP (μm s−1)68.39 ± 3.3a50.05 ± 2.0b0.000161.24 ± 3.8a51.45 ± 2.9b0.00473.60 ± 4.585.75 ± 4.20.067
VSL (m s−1)46.78 ± 2.3a37.75 ± 1.7b0.01244.96 ± 3.240.41 ± 2.60.08882.29 ± 5.792.39 ± 5.40.220
LIN (%)33.2 ± 1.035.40 ± 0.70.24736.6 ± 0.938.21 ± 0.70.074108.76 ± 6.1104.82 ± 2.30.505
STR (%)67.9 ± 1.6a74.90 ± 1.3b0.03972.43 ± 1.1a77.57 ± 1.1b0.001113.22 ± 5.5107.33 ± 1.80.261
WOB (%)48.43 ± 0.947.30 ± 0.40.59850.36 ± 0.845.79 ± 3.20.16596.22 ± 0.990.97 ± 6.30.498
BCF (Hz)24.86 ± 0.524.43 ± 0.70.74026.92 ± 0.626.99 ± 0.70.915100.45 ± 2.7100.49 ± 2.50.792
ALH (μm)5.22 ± 0.2a4.10 ± 0.16b0.0054.35 ± 0.3a3.66 ± 0.1b0.00179.13 ± 4.986.12 ± 3.40.241

ALH: amplitude of the lateral head displacement; BCF: beat cross frequency; CT: cryotolerance, LIN: linearity, PM: progressive motility, SE: standard error of mean; STR: straightness, TM: total motility, VAP: average path velocity; VCL; curvilinear velocity; VSL: straight line velocity, WOB: wobble.

Means in the same row with different superscript within a breeda,b (Merino or Racka) differ significantly.

Testicles number: Merino (n = 14), Racka (n = 14).

The interaction effects of the collection methods and breeds on motility and kinematic parameters of epididymal spermatozoa from Merino and Racka rams

Table 4 presents the interaction effects of the collection methods and breeds on the motility and kinematic parameters of Merino and Racka ram epididymal spermatozoa. There was no significant (P > 0.05) collection methods and breed effects on the motility and all the kinematic parameters of Merino and Racka ram epididymal spermatozoa.

Table 4.

Breed and collection method interaction effects on motility and kinematic parameters of Merino and Racka ram epididymal spermatozoa under fresh and post-thaw conditions

ParametersForm (Mean ± SE)
FreshP-valuePost-thawP-value
Collection methodsMerinoRackaMerinoRacka
TM (%)SL85.29 ± 3.778.86 ± 3.70.73442.60 ± 7.948.14 ± 6.70.163
IN82.00 ± 3.778.14 ± 3.745.80 ± 7.930.14 ± 6.7
PM (%)SL77.14 ± 4.568.86 ± 4.50.36028.00 ± 7.934.14 ± 6.70.235
IN68.86 ± 4.569.00 ± 4.532.00 ± 7.920.14 ± 6.7
VCL (μm s−1)SL141.14 ± 12.5124.09 ± 12.50.906106.20 ± 8.9100.26 ± 7.50.533
IN139.14 ± 12.5119.07 ± 12.5103.33 ± 8.9107.93 ± 7.5
VAP (μm s−1)SL70.03 ± 5.262.46 ± 5.20.93650.70 ± 4.350.20 ± 3.60.637
IN66.74 ± 5.260.03 ± 5.249.41 ± 4.352.70 ± 3.6
VSL (m s−1)SL47.90 ± 4.146.17 ± 4.10.98237.80 ± 3.839.67 ± 3.20.824
IN45.66 ± 4.143.74 ± 4.137.70 ± 3.841.16 ± 3.2
LIN (%)SL33.86 ± 1.437.29 ± 1.41.00035.00 ± 1.039.14 ± 0.90.181
IN32.57 ± 1.436.00 ± 1.435.80 ± 1.037.29 ± 0.9
STR (%)SL68.43 ± 2.073.29 ± 2.00.88874.00 ± 1.878.29 ± 1.50.348
IN67.29 ± 2.071.57 ± 2.075.80 ± 1.876.86 ± 1.5
WOB (%)SL49.43 ± 1.250.43 ± 1.20.45447.20 ± 4.243.00 ± 3.50.436
IN47.43 ± 1.250.29 ± 1.247.40 ± 4.248.57 ± 3.5
BCF (Hz)SL25.16 ± 0.827.21 ± 0.81.00023.94 ± 1.227.36 ± 1.00.494
IN24.57 ± 0.826.63 ± 0.824.92 ± 1.226.63 ± 1.0
ALH (μm)SL5.33 ± 0.44.39 ± 0.40.8324.23 ± 0.23.59 ± 0.20.393
IN5.09 ± 0.44.30 ± 0.43.98 ± 0.23.73 ± 0.2

ALH: amplitude of the lateral head displacement; BCF: beat cross frequency; LIN: linearity, PM: progressive motility, SE: standard error of mean; STR: straightness; TM: total motility, VAP: average path velocity; VCL; curvilinear velocity; VSL: straight line velocity, WOB: wobble.

Discussion

Epididymal spermatozoa (EPS) are an excellent alternative for collecting and freezing gametes from domestic and wild animals for gene conservation post mortem. The efficiency of retrieving high quality EPS is also of crucial importance in the case of late processing. The motility and concentration of spermatozoa are two main factors that determine sperm function (effective fertilization), with movement characteristics being the central component (Robayo et al., 2008). The purpose of the current study was to assess the effects of two EPS recovery methods (slicing vs incision) on the movement characteristics of fresh and frozen-thawed Merino and Racka REPS's and to compare the fresh semen quality and cryotolerance in the two breeds. The weight of the testicles between the collection methods was not significantly different, ensuring that any differences observed are not attributable to the variation in the testicular weight. The collection method did not significantly affect the standard motility and kinematic parameters of the fresh and post-thaw REPS. However, the IN method was faster as it does not require the removal of the tunica vaginalis and thus is more practical in field conditions. In contrast, we found it quite challenging to establish a total cell number with the SL method due to wide variations in the number of cells retrieved. This is in line with the report of Ehling et al. (2006). Our findings tallies with the findings of Martínez-Pastor et al. (2006) who have reported that the collection method (cuts vs flushing) did not significantly affect the motility, total number and post-thaw results of EPS recovered from Iberian red deer. Similarly, Kang et al. (2018) have found no significant (P > 0.05) difference between the post-thaw standard motility parameters of EPS obtained from Hanwoo bull by the flushing or mincing methods. In contrast, in rams, the IN method gave a sample with significantly (P > 0.05) higher total motility than the mincing method (75.0 vs 59.2) (Lone et al., 2011). Our results are in agreement with the findings of Mogheiseh et al. (2022), who have reported that the EPS retrieval methods have no significant effects on the post-thaw standard motility and kinematic parameters of dog EPS.

The Merino breed presented significantly (P > 0.05) higher testicular weight than the Racka breed (260.50 ± 11.8 vs 170.82 ± 10.1 g). This might be due to the testicular size/scrotal circumference difference or body weight between the breeds (Allaoui et al., 2014). Moreover, it was established that ram testicular diameter and scrotal circumferences were significantly (P < 0.01) correlated with testicular weight (Endale et al., 2009). The lowest and the highest fresh TM and PM recorded in the current study (78.50 ± 2.8 to 83.64 ± 2.2%) and (68.93 ± 3.3 to 73.00 ± 3.1%) were slightly higher than that of Kaabi et al. (2003), 78.4 and 64.7% retrieved 24 h after animal death but lower than what was reported by Rahimizadeh et al. (2021), 95.89 and 90.40% (recovered few hours after the animals were slaughtered), respectively. This might be due to the differences in the REPS retrieval season and time past after slaughtering. Bergstein-Galan et al. (2018) reported that the PM and viability of REPS declined significantly at 12 and 48 h after the animal death, respectively.

We found that the breed has no significant (P > 0.05) effects on the fresh TM and PM. This is in contrast with the result of Vozaf et al. (2022) who have compared Slovak Dairy, Native Wallachian and Improved Wallachian breeds. Similarly, Kasimanickam et al. (2007) have reported that the progressive motility of chilled-stored electro-ejaculated ram spermatozoa were different in case of breeds Polled Dorset, Suffolk and Katahdin. The EPS of the Racka breed were moving straighter than the EPS of Merino, as indicated by their higher LIN, STR and lower ALH values. This contrasted with the results by Kasimanickam et al. (2007), according to which breed does not have significant effects on the kinematic parameters of chilled stored electro-ejaculated ram spermatozoa. Additionally, the spermatozoa of both breeds were rapid as sperm cells with a VAP value over 50 μm s−1 have been categorized rapid-moving spermatozoa (Goovaerts et al., 2006).

After thawing, none of the standard motility parameters differed significantly (P > 0.05) between the breeds as in the case of fresh sperm. The post-thaw TM of Merino (44.20 ± 6.47%) was lower than that (50.9 ± 3.1%) reported by Çoyan et al. (2011) for AV-collected spermatozoa of Merino. However, we recorded a higher PM of 30.00 ± 6.97% vs 19.6 ± 1.1%. In Racka, we obtained lower values for TM (39.14 ± 4.48%) and PM (27.14 ± 3.82%) than those (60.86 ± 5.44% and 52.86 ± 6.12%, respectively) reported for the same breed previously (Egerszegi et al., 2012). This might be due to the difference in the collection seasons. Our results agree with those of Vozaf et al. (2022) who have compared three breeds and found that the ram breed does not have significant effects (P > 0.05) on the post-thaw standard motility parameters. To the contrary, Tohura et al. (2019) have reported that the breed had significant effects (P < 0.05) on the motility of bull spermatozoa. Similarly, most of the differences in the kinematic parameters of the two breeds remain the same as in the fresh except for STR, which was significantly higher in Racka in the fresh and not significantly different between the breed after thawing. In addition, the Racka breed presented significantly (P < 0.05) higher LIN and BCF values in both fresh and post-thaw kinematic parameters than the Merino breed. This shows that EPS of the Racka breed had better linear movement than those of the Merino. Higher sperm swimming velocities determine male fertilizing ability (Malo et al., 2006), while higher BCF and lower ALH of the sperm head could facilitate the penetration of the zona pellucida (Goovaerts et al., 2006). Similarly, LIN was reported to impact positively the cleavage rate following IVF and is a significant predictor of cleavage rate (García-Alvarez et al., 2009).

Freezing and thawing affected the same parameters in both breeds except for VSL, which was not significantly affected in the Racka (P > 0.05). Similarly, the REPS of both breeds had similar cryotolerance. Our results contradict those published by Andreeva et al. (2017), who have compared the Bulgarian dairy synthetic population with the Ile de France breed and recorded a significant difference in the cryotolerance of the spermatozoa.

Our findings revealed that the collection method does not influence significantly the standard motility and kinematic parameters of the fresh or post-thaw ram epididymal spermatozoa. The incision method seems faster and more field-friendly than the slicing method. However, if the visceral layer of the tunica vaginalis communis contains larger blood vessels, removal of the tunica and the slicing method is recommended to avoid bloody sperm samples. We observed that the Racka EPS, either in fresh or post-thaw conditions have straighter movement than those of the Merinos. We found the cryotolerance of REPS from both breeds identical.

In literature, diverse motility parameters are described, however, it is well documented that the kinematic parameters of the sperm are insufficient to predict the fertilizing ability of the semen. Further investigation are needed for a more profound evaluation of REPS by the analysis of the possible effects of the side and position (right or left) of the epididymis of a given animal on the post-thaw quality of sperm. Similarly, the influence of the dilution rate and the freezing extenders should also be studied in more detail.

Acknowledgement

This study was supported by the Stipendium Hungaricum Scholarship Program,  award number (SHE-07824-004/2019). The authors wish to thank Judit Radnainé Szentpáli for her assistance in the laboratory work and the Orosz és Tsa Ltd. Hungary, for providing the testicles.

References

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    • Search Google Scholar
    • Export Citation
  • Allaoui, A., Safsaf, B., Laghrour, W. and Tlidjane, M. (2014): Factors affecting scrotal measurements and weight of Ouled Djellal rams in Eastern and South-eastern Algeria. APCBEE Procedia 8 ,260265. https://doi.org/10.1016/j.apcbee.2014.03.037.

    • Search Google Scholar
    • Export Citation
  • Alvarez, M., Tamayo-Canul, J., Anel, E., Boixo, J. C., Mata-Campuzano, M., Martinez-Pastor, F., Anel, L. and de Paz, P. (2012): Sperm concentration at freezing affects post-thaw quality and fertility of ram semen. Theriogenology 77 ,11111118. https://doi.org/10.1016/j.theriogenology.2011.10.013.

    • Search Google Scholar
    • Export Citation
  • Amir, D. and Volcani, R. (1965): Seasonal fluctuations in the sexual activity of Awassi, German Mutton Merino, Corriedale, Border-Leicester and Dorset Horn rams: I. Seasonal changes in semen plasma volume and its fructose and citric acid concentrations. The J. Agric. Sci. 64, 115120. https://doi.org/10.1017/S0021859600010558.

    • Search Google Scholar
    • Export Citation
  • Andreeva, M. N., Metodiev, N. T., Tsvetkov T. S. and Stefanov, R. G. (2017): The influence of the sheep breed on the cryotolerance of spermatozoa. In: Proceedings of the International Scientific Conference “Veterinary Medicine in Service of People”, 6–7 October 2017, Stara Zagora, Bulgaria.

    • Search Google Scholar
    • Export Citation
  • Anon. (2023). Racka Sheep: Characteristics and Best 23 Facts. Retrieved July 25, 2023 from: https://www.roysfarm.com/racka-sheep/.

  • Arando, A., Delgado, J. V., León, J. M., Nogales, S., Navas-González, F. J., Pizarro, M. G. and Pérez-Marín, C. C. (2019): Effect of three commercial extenders on sperm motility and fertility in liquid ram semen stored at 15oC or 5oC. Acta Vet. Hung. 67, 430444. https://doi.org/10.1556/004.2019.043.

    • Search Google Scholar
    • Export Citation
  • Bergstein-Galan, T. G., Weiss, R. R., Barbosa, T. S. R., Kozicki, L. E. and Bicudo, S. D. (2018): Viability of ovine spermatozoa collected from epididymides stored at 18°-25°C for 48 hours post mortem. Arq. Bras. Med. Vet. Zootec. 70 ,10231028. https://doi.org/10.1590/1678-4162-10058.

    • Search Google Scholar
    • Export Citation
  • Bertol, M. A. F. (2016): Cryopreservation of epididymal sperm. In: Marco-Jimenez, F. and Akdemir, H. (eds) Cryopreservation in Eukaryotes. InTech. https://doi.org/10.5772/65010.

    • Search Google Scholar
    • Export Citation
  • Çoyan, K., Başpınar, N., Bucak, M. N. and Akalın, P. P. (2011): Effects of cysteine and ergothioneine on post-thawed Merino ram sperm and biochemical parameters. Cryobiology 63, 16. https://doi.org/10.1016/j.cryobiol.2011.04.001.

    • Search Google Scholar
    • Export Citation
  • DAD-IS. (2020): Domestic Animal Diversity Information System. Retrieved July 21, 2023, from: https://www.fao.org/dad-is/sdg-252/en/.

  • Ehling, C., Rath, D., Struckmann, C., Frenzel, A., Schindler, L. and Niemann, H. (2006): Utilization of frozen-thawed epididymal ram semen to preserve genetic diversity in Scrapie susceptible sheep breeds. Theriogenology 66 ,21602164. https://doi.org/10.1016/j.theriogenology.2006.07.003.

    • Search Google Scholar
    • Export Citation
  • Egerszegi, I., Sarlos, P. and Ratky, J. (2012): Cryopreservation of epididymal spermatozoa of Black Racka rams from Hortobágy – a possible means of gene preservation. [in Hungarian] Magy. Allatorvosok 134, 524528. https://www.cabdirect.org/cabdirect/abstract/20123328495.

    • Search Google Scholar
    • Export Citation
  • Endele, M., Hailemariam, M. and Tegegne, A. (2009): A study on testicular characteristics of ram lambs of Arsi breed fed on two maize varieties (QPM and BH540). Ethiop. J. Sci. 32, 3744. https://doi.org/10.4314/sinet.v32i1.67877.

    • Search Google Scholar
    • Export Citation
  • Fernández Abella, D., Da Costa, M., Guérin, Y. and Dacheux, J. L. (2015): Fertility of undiluted ram epididymal spermatozoa stored for several days at 4°C. Animal 9 ,313319. https://doi.org/10.1017/S1751731114002109.

    • Search Google Scholar
    • Export Citation
  • García-Alvarez, O., Maroto-Morales, A., Martínez-Pastor, F., Garde, J. J., Ramón, M., Fernández-Santos, M. R., Esteso, M. C., Pérez-Guzmán, M. D., and Soler, A. J. (2009): Sperm characteristics and in vitro fertilization ability of thawed spermatozoa from Black Manchega ram: electroejaculation and postmortem collection. Theriogenology 72 ,160-168. https://doi.org/10.1016/j.theriogenology.2009.02.002.

    • Search Google Scholar
    • Export Citation
  • Goovaerts, I. G. F., Hoflack, G. G., Van Soom, A., Dewulf, J., Nichi, M., de Kruif, A. and Bols, P. E. J. (2006): Evaluation of epididymal semen quality using the Hamilton–Thorne analyser indicates variation between the two caudae epididymides of the same bull. Theriogenology 66 ,323330. https://doi.org/10.1016/j.theriogenology.2005.11.018.

    • Search Google Scholar
    • Export Citation
  • Kaabi, M., Paz, P., Alvarez, M., Anel, E., Boixo, J. C., Rouissi, H., Herraez, P. and Anel, L. (2003): Effect of epididymis handling conditions on the quality of ram spermatozoa recovered post-mortem. Theriogenology 60 ,12491259.

    • Search Google Scholar
    • Export Citation
  • Kang, S.-S., Kim, U.-H., Jeon, M.-H., Lee, M.-S. and Cho, S.-R. (2018): Comparison of spermatozoa recovery methods on cauda epididymal sperm of Hanwoo bulls. J. Anim. Reprod. Biotech. 33, 321326. https://doi.org/10.12750/JET.2018.33.4.321.

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  • Karja, N. K. W., Respaty, E. M. A., Nuraini, I., Prihatno, S. A. and Gustari, S. (2010): Characteristic of frozen-thawed epididymal spermatozoa and refrigerated storage of ram spermatozoa. J. Indones. Trop. Anim. Agric. 35 ,6367. https://doi.org/10.14710/jitaa.35.1.63-67.

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  • Kasimanickam, R., Kasimanickam, V., Pelzer, K. D. and Dascanio, J. J. (2007): Effect of breed and sperm concentration on the changes in structural, functional and motility parameters of ram-lamb spermatozoa during storage at 4°C. Anim. Reprod. Sci. 101 ,6073. https://doi.org/10.1016/j.anireprosci.2006.09.001.

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  • Lone, F. A., Islam, R., Khan, M. Z. and Sofi, K. A. (2011): Effect of collection methods on the quality and quantity of spermatozoa recovered from the cauda epididymidis of slaughtered ram. Indian Vet. J. 88, 46. https://www.researchgate.net/publication/289314785_.

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  • Martínez-Pastor, F., Martínez, F., García-Macías, V., Esteso, M. C., Anel, E., Fernández-Santos, M. R., Soler, A. J., de Paz, P., Garde, J. and Anel, L. (2006): A pilot study on post-thawing quality of Iberian red deer spermatozoa (epididymal and electro-ejaculated) depending on glycerol concentration and extender osmolality. Theriogenology 66 ,1165-1172. https://doi.org/10.1016/j.theriogenology.2006.03.027.

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  • Mogheiseh, A., Divar, M. R., Vesal, N. and Mahdivand Moradloo, F. (2022): Effects of epididymal sperm recovery methods on fresh and frozen–thawed sperm characteristics in dogs. Reprod. Domest. Anim. 57, 10381045. https://doi.org/10.1111/rda.14171.

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  • Mujitaba, M. A., Egerszegi, I., Kútvölgyi, G., Nagy, S., Vass, N. and Bodó, S. (2022): Alternative opportunities to collect semen and sperm cells for ex situ in vitro gene conservation in sheep. Agriculture 12 ,2001. https://doi.org/10.3390/agriculture12122001.

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  • Ocampo, L. C., DelaRosa, I. C. J., Lofranco, J. O. C., Ortiz, J. G. M. and Ocampo, M. B. (2021): Live birth after artificial insemination using cryopreserved epididymal sperm recovered from the cauda epididymis of slaughtered non-descript bucks in the Philippines. Int. J. Agric. Tech. 17 ,21832196.

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  • Parra-Forero, L. Y., Mendoza, G. D., Góngora, A., López Fernández, M. D. C., Cruz, L. A., Montiel, A. J., Kjelland, M. E. and García-Contreras, A. D. C. (2015): Azteca Breed horse epididymal sperm evaluation: a Comparison of head, corpus and cauda sperm quality. Advan. Reprod. Sci. 3 ,5765.

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  • Rahimizadeh, P., Rezaei Topraggaleh, T., Bucak, M. N., Ziarati, N., Hasirbaf, A., Taher-Mofrad, S. M. J., Maroufizadeh, S. and Shahverdi, A. (2021): Effect of bovine serum albumin supplementation in tris-soybean lecithin-based extender on quality of chilled ram epididymal spermatozoa. Biopreserv. Biobank. 19 ,3340. https://doi.org/10.1089/bio.2020.0041.

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  • Rickard, J. P., Pini, T., Soleilhavoup, C., Cognie, J., Bathgate, R., Lynch, G. W., Evans, G., Maxwell, W. M. C., Druart, X. and de Graaf, S. P. (2014): Seminal plasma aids the survival and cervical transit of epididymal ram spermatozoa. Reproduction 148 ,469478. https://doi.org/10.1530/REP-14-0285.

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  • Vozaf, J., Svoradová, A., Baláži, A., Vašíček, J., Olexiková, L., Dujíčková, L., Makarevich, A. V., Jurčík, R., Ďúranová, H. and Chrenek, P. (2022): The cryopreserved sperm traits of various ram breeds: towards biodiversity conservation. Animals 12 ,1311. https://doi.org/10.3390/ani12101311.

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  • Zsolnai, A., Egerszegi, I., Rózsa, L. and Anton, I. (2021): Genetic status of lowland-type Racka sheep colour variants. Animal 15, 100080. https://doi.org/10.1016/j.animal.2020.100080.

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Senior editors

Editor-in-Chief: Ferenc BASKA

Editorial assistant: Szilvia PÁLINKÁS

 

Editorial Board

  • Mária BENKŐ (Acta Veterinaria Hungarica, Budapest, Hungary)
  • Gábor BODÓ (University of Veterinary Medicine, Budapest, Hungary)
  • Béla DÉNES (University of Veterinary Medicine, Budapest Hungary)
  • Edit ESZTERBAUER (Veterinary Medical Research Institute, Budapest, Hungary)
  • Hedvig FÉBEL (National Agricultural Innovation Centre, Herceghalom, Hungary)
  • László FODOR (University of Veterinary Medicine, Budapest, Hungary)
  • János GÁL (University of Veterinary Medicine, Budapest, Hungary)
  • Balázs HARRACH (Veterinary Medical Research Institute, Budapest, Hungary)
  • Peter MASSÁNYI (Slovak University of Agriculture in Nitra, Nitra, Slovak Republic)
  • Béla NAGY (Veterinary Medical Research Institute, Budapest, Hungary)
  • Tibor NÉMETH (University of Veterinary Medicine, Budapest, Hungary)
  • Zsuzsanna NEOGRÁDY (University of Veterinary Medicine, Budapest, Hungary)
  • Dušan PALIĆ (Ludwig Maximilian University, Munich, Germany)
  • Alessandra PELAGALLI (University of Naples Federico II, Naples, Italy)
  • Kurt PFISTER (Ludwig-Maximilians-University of Munich, Munich, Germany)
  • László SOLTI (University of Veterinary Medicine, Budapest, Hungary)
  • József SZABÓ (University of Veterinary Medicine, Budapest, Hungary)
  • Péter VAJDOVICH (University of Veterinary Medicine, Budapest, Hungary)
  • János VARGA (University of Veterinary Medicine, Budapest, Hungary)
  • Štefan VILČEK (University of Veterinary Medicine in Kosice, Kosice, Slovak Republic)
  • Károly VÖRÖS (University of Veterinary Medicine, Budapest, Hungary)
  • Herbert WEISSENBÖCK (University of Veterinary Medicine, Vienna, Austria)
  • Attila ZSARNOVSZKY (Szent István University, Gödöllő, Hungary)

ACTA VETERINARIA HUNGARICA
Institute for Veterinary Medical Research
Centre for Agricultural Research
Hungarian Academy of Sciences
P.O. Box 18, H-1581 Budapest, Hungary
Phone: (36 1) 287 7073 (ed.-in-chief) or (36 1) 467 4081 (editor)

E-mail: acta.veterinaria@univet.hu (ed.-in-chief)

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2022  
Web of Science  
Total Cites
WoS
972
Journal Impact Factor 0.900
Rank by Impact Factor

Veterinary Sciences 95/143

Impact Factor
without
Journal Self Cites
0.900
5 Year
Impact Factor
1.1
Journal Citation Indicator 0.47
Rank by Journal Citation Indicator

Veterinary Sciences 103/170

Scimago  
Scimago
H-index
38
Scimago
Journal Rank
0.277
Scimago Quartile Score

Veterinary (miscellaneous) Q2

Scopus  
Scopus
Cite Score
1.9
Scopus
CIte Score Rank
General Veterinary 76/186 (59th PCTL)
Scopus
SNIP
0.475

2021  
Web of Science  
Total Cites
WoS
1040
Journal Impact Factor 0,959
Rank by Impact Factor Veterinary Sciences 103/144
Impact Factor
without
Journal Self Cites
0,876
5 Year
Impact Factor
1,222
Journal Citation Indicator 0,48
Rank by Journal Citation Indicator Veterinary Sciences 106/168
Scimago  
Scimago
H-index
36
Scimago
Journal Rank
0,313
Scimago Quartile Score Veterinary (miscellaneous) (Q2)
Scopus  
Scopus
Cite Score
1,7
Scopus
CIte Score Rank
General Veterinary 79/183 (Q2)
Scopus
SNIP
0,610

2020  
Total Cites 987
WoS
Journal
Impact Factor
0,955
Rank by Veterinary Sciences 101/146 (Q3)
Impact Factor  
Impact Factor 0,920
without
Journal Self Cites
5 Year 1,164
Impact Factor
Journal  0,57
Citation Indicator  
Rank by Journal  Veterinary Sciences 93/166 (Q3)
Citation Indicator   
Citable 49
Items
Total 49
Articles
Total 0
Reviews
Scimago 33
H-index
Scimago 0,395
Journal Rank
Scimago Veterinary (miscellaneous) Q2
Quartile Score  
Scopus 355/217=1,6
Scite Score  
Scopus General Veterinary 73/183 (Q2)
Scite Score Rank  
Scopus 0,565
SNIP  
Days from  145
submission  
to acceptance  
Days from  150
acceptance  
to publication  
Acceptance 19%
Rate

 

2019  
Total Cites
WoS
798
Impact Factor 0,991
Impact Factor
without
Journal Self Cites
0,897
5 Year
Impact Factor
1,092
Immediacy
Index
0,119
Citable
Items
59
Total
Articles
59
Total
Reviews
0
Cited
Half-Life
9,1
Citing
Half-Life
9,2
Eigenfactor
Score
0,00080
Article Influence
Score
0,253
% Articles
in
Citable Items
100,00
Normalized
Eigenfactor
0,09791
Average
IF
Percentile
42,606
Scimago
H-index
32
Scimago
Journal Rank
0,372
Scopus
Scite Score
335/213=1,6
Scopus
Scite Score Rank
General Veterinary 62/178 (Q2)
Scopus
SNIP
0,634
Acceptance
Rate
18%

 

Acta Veterinaria Hungarica
Publication Model Hybrid
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Acta Veterinaria Hungarica
Language English
Size A4
Year of
Foundation
1951
Volumes
per Year
1
Issues
per Year
4
Founder Magyar Tudományos Akadémia
Founder's
Address
H-1051 Budapest, Hungary, Széchenyi István tér 9.
Publisher Akadémiai Kiadó
Publisher's
Address
H-1117 Budapest, Hungary 1516 Budapest, PO Box 245.
Responsible
Publisher
Chief Executive Officer, Akadémiai Kiadó
ISSN 0236-6290 (Print)
ISSN 1588-2705 (Online)

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