Research Article

Assessment of the Quality and Performance of Jersey and Holstein Frozen Semen in Gandaki Province, Nepal

Rupak Kandel^1*, Sanjay Paudel², Khagendra Raj Sapkota³, Jagadish Pandaya⁴, Dawa Tshiring Tamang⁵

¹Faculty of Animal Science, Veterinary Science and Fisheries, Agriculture and Forestry University, Bharatpur, Nepal

²Institute of Agriculture and Animal Science, Tribhuvan University, Paklihawa, Rupandehi, Nepal

³National Livestock Breeding Office, Department of Livestock Services, Nepaljung, Nepal

⁴National Livestock Breeding Office, Department of Livestock Services, Pokhara, Nepal

⁵Department of Animal Breeding and Biotechnology, Agriculture and Forestry University (AFU), Rampur, Chitwan, Nepal

Submission: June 3, 2025; Published: June 13, 2025

*Corresponding author: Rupak Kandel, Faculty of Animal Science, Veterinary Science and Fisheries, Agriculture and Forestry University, Bharatpur, Nepal

How to cite this article: Rupak K, Sanjay P, Khagendra Raj S, Jagadish P, Dawa Tshiring T. Assessment of the Quality and Performance of Jersey and 002 Holstein Frozen Semen in Gandaki Province, Nepal. Arch Anim Poult Sci. 2025; 3(1): 555604.DOI: 10.19080/AAPS.2025.03.555604

Abstract

With its 24.01% AGDP contribution and 16% contribution to the nation’s overall protein demand, livestock is essential to Nepal’s rural livelihoods. Livestock is still essential for economic and nutritional stability even if the numbers of cattle, buffalo, sheep, and chickens are declining. The Agriculture Development Strategy (ADS-2015-2035) focuses on livestock advances, especially genetic upgrades, with the goal of reducing hunger. The purpose of this study is to assess the effectiveness and quality of frozen semen from bulls raised in the areas of Nawalpur, Parbat, Baglung, and Myagdi that are Jersey and Holstein. The primary goal is to evaluate the quality of semen used in artificial insemination (AI) to improve genetics and increase productivity and production. Analyzing sperm concentration, motility, and viability, comparing semen efficiency across various geographic regions, and looking at factors impacting semen efficacy are some of the specific goals. Semen quality parameters such as volume, density, concentration, motility, and viability were assessed using standard protocols. Semen from Jersey and Holstein bulls were collected and analyzed for its potential to enhance genetic improvement in livestock.

The study revealed significant variations in semen quality across different breeds and geographical areas. Jersey and Holstein bulls exhibited higher motility and viability. Factors such as storage conditions and handling practices were found to impact semen quality significantly. There was no significant difference in species and concentration of sperm for frozen semen although the highest concentration was found in Jersey semen followed by Holstein. The highest post-thaw motility was observed in Jersey semen followed by Holstein semen. There was no significant difference between the species and post-thaw motility of frozen semen. The highest viability in the 3rd hour was found to be for Jersey as compared to Holstein. There was significant difference between the breed and viability of sperm. Given that the acrosome includes enzymes essential for the penetration of the zona pellucida and the surrounding cellular layers, freezing and thawing procedures have the potential to seriously harm the membrane structures of the sperm head. According to reports, acrosome integrity has a stronger correlation with frozen bull semen fertility than motility. However, sperm motility plays a crucial role in the processes that lead to gamete contact. A decrease in post-thaw sperm velocity and in the percentage of motile cells that are advancing indicates cellular damage that may lessen the likelihood of fertilization.

Keywords:Sperm Concentrations; Motility; Viability; Breed; Motile Cells

Abbreviation:ADS: Agriculture Development Strategy; AI: Artificial Insemination; AGDP: Annual Gross Domestic Product; MoALD: Ministry of Agriculture and Livestock Development; DLS: Department of Livestock Services; MT: Metric Ton; BMI: Body Mass Index; HF: Holstein Freisian; NLBO: National Livestock Breeding Office; ml: Milliliter; DCIP: Dairy Cattle Improvement Project; PPRS: Pedigree Performance Recording Scheme

Introduction

In Nepal, resilient rural livelihoods require livestock. Livestock provides about 16% of the total protein required and contributes 24.01% of the AGDP of the nation. Even though the fiscal year 2079/080’s livestock comprises more goats and ducks than in previous years, the population of cattle, buffalo, sheep and swine dropped significantly. There are now 35.9% fewer cows and 39.9% less buffaloes, respectively. The number of sheep, swine and poultry has decreased by 34.9%, 9.77%, and 2.39% respectively (MoALD, 2022). Presently, cattle contribute about 46.44% to total milk production of 26,13,843 MT per year nationwide (DLS, 2081). Limiting stunting, underweight, and wasting of women and children with low body mass index (BMI) to 8, 5, 1, and 5%, respectively, by 2035 is envisioned in the Agriculture Development Strategy (ADS-2015-2035), a major policy guideline for agriculture and livestock development in Nepal. This goal is primarily dependent on livestock innovations. Ruminants serve a variety of economic, social, and risk-reduction purposes, making them an invaluable asset for low-income households [1]. A disproportionately high proportion of tiny ruminants are native breeds connected to marginalized groups and habitats.

The dairy cows are crossbreds of indigenous cattle with Holstein or Jersey, meaning they have diverse genetic constitutions. The ideal degree of crossbreeding for the various agroclimatic conditions in Nepal’s various areas is not well understood scientifically [2]. While Jersey cattle and their crossbred cows have higher fat content and are more robust, making them more suitable for resource-poor areas, Holstein Frisian (HF) cattle and their crossbred cattle have higher milk yields with lower fat and protein contents and require good management and high-quality feeding [3].

It is generally anticipated that non-genetic factors will influence production potential and contribute to productive performance. Understanding these aspects enables a more precise evaluation of breeding values. In this regard, characteristics of livestock’s reproductive and productive capacities and the factors influencing them have not been thoroughly studied, with a greater emphasis on improved genetic performance in dairy cattle [4].

However, information regarding handling and efficacy of frozen semen, particularly in western Nepal, where there is a dispersed population and no selection plan, has been recorded minimum. One of the Government of Nepal’s top priorities for raising the productivity of the country’s livestock herd is genetic enhancement. Performance evaluations of the improved livestock herd at the field level are rare. The Jersey and Holstein bulls utilized in Artificial Insemination (AI) do not have an adequate or nonexistent assessment mechanism in place. All of these bulls ought to be assessed based on how well their semen is conceived, as this is a factor in the AI program [5]. Artificial Insemination is a commonly used technology for enhancing livestock species’ genetic potential. In farmers’ fields, Nepalese livestock have been updated with varying degrees of foreign blood (Jersey and HF), with varying potentialities for productivity. Exotic breeds from temperate climates are not suited to tropical circumstances.

The reproductive effectiveness of Friesian and Jersey bulls can be negatively impacted by these conditions, therefore it’s important to regularly assess the quality and quantity of their semen to increase non-return rates and maintain the crossbreeding program’s financial viability. After 50 years of use in the nation, it is now vital to determine how AI has improved the breed of Nepalese livestock and to identify the genotype and phenotype of the Nepalese Jersey, and Holstein breed that would yield the best degree of output performance.

This study aims to evaluate and improve future genetic potential and productivity by examining the efficacy of frozen semen in Jersey, Holstein and Murrah in Nawalpur, Parbat, Baglung and Myagdi districts, as well as the factors influencing these performances under farmer management.

Materials and Methods

To assess the amount and quality of frozen cattle bull semen utilized for AI programs in cattle across the nation, a study was conducted at the National Livestock Breeding Office (NLBO), Pokhara. The quantity, quality, and status of the semen produced by Jersey and Holstein bulls were assessed. Bull semen was examined physically and microscopically in accordance with this. The frozen semen samples from AI technicians were collected from Nawalpur, Parbat, Baglung and Myagdi districts of western Nepal and their efficacy was monitored. The study to assess the quantity and quality of frozen Jersey, Holstein and Murrah bull semen utilized in cattle AI program across the Gandaki Province was made from February to May of 2024. The study site and sample analysis site are shown in the (figure 1) below.

The quality of semen produced by Jersey and Holstein bulls were assessed, along with their status. Finally, breed-to-breed and district-by-district comparisons of the effectiveness of frozen semen were made.

To assess the quality of bull semen, the following metrics were noted.

I. Mass activity of sperms: Sperm motility in a bulk or group

II. Initial motility of sperms: Percentage of each fresh semen sperm’s initial motility

III. Sperm concentration: Million sperm/ml (106 /ml) of semen

IV. Doses of semen generated: Doses of semen straw made during semen ejaculation.

Breed type, lactation milk output, and monthly milk yield of a selected animal that included protein, fat, and value solids will be derived from the Dairy Cattle Improvement Project (DCIP) of Nepal’s Pedigree Performance Recording Scheme (PPRS). At NLBO in Pokhara, PPRS data from the DCIP were recorded and examined. The efficacy of frozen semen was evaluated as represented in sample (figure 2) and analyzed based on the quality of semen presented in (table 1).

Semen straw dispatched to AI technicians were collected from Nawalpur, Parbat, Baglung and Myagdi districts of Gandaki Province. The samples were collected, kept in Nitrogen tank maintained at -196°C and transferred to NLBO laboratory, Pokhara. In accordance with the standard operating procedure for Bovine Frozen Semen Production issued by NLBO, Pokhara and approved by DLS, tests on semen concentration, and motility as well as post-thaw motility and post-thaw viability were carried out at the quality control unit of NLBO, Pokhara. Using a photometer, the concentration of semen was determined. After placing a drop of fresh semen onto a glass slide, the motility of the semen was manually measured using a microscope. The frozen semen straw was then thawed, and a drop of thawed semen was placed in a glass slide to manually evaluate post-thaw motility under a microscope. Data was analyzed by the least squares approach using Harvey (1990) software package to explore the major cause of variation and effect of non-genetic factors on productive and reproductive qualities and to address the challenge of disproportionate subclass numbers. We computed the coefficient of variation, standard error of mean, and overall mean using the SPSS 20.0 software package. One way ANOVA was used to compare the statistically significant means. MS Excel 2013 was used to analyze manufacturing system data.

Results and Discussion

Bull semen’s physical quality, or macroscopic quality, and microscopic quality are its quality parameters. The color, volume, and density of bull semen are among its physical quality criteria. The present investigation examined the microscopic quality characteristics of bull semen, including initial motility, sperm concentration, post-thaw motility, and sperm viability.

Microscopic Quality Parameter of Bull Semen

Concentration of the Sperm

Jersey bull semen had an overall sperm concentration of 215.40±65.17 (105/ml). Similarly, the sperm concentration from Holstein bull semen was found to be 192.80±58.08 (105/ml). For both beef and dairy bull semen, the mean concentration (±SD) was 880±20 (105/ml) and 550±19 (105/ml), respectively (P<0.001) [5] shown in (table 2).

There was no significant difference in species and concentration of sperm for frozen semen although the highest concentration was found in Jersey semen i.e. 215.40 (10⁵/ml) followed Holstein bulls 192.80 (10⁵/ml). Similarly, concentration of sperm from three geographical regions i.e. Baglung-Myagdi, Parbat and Nawalpur were found to be 208.57±62.32, 210.25±70.63 and 220.38±43.66 (10⁵/ml) respectively.

There was no significant difference in geographical area and concentration of sperm for frozen semen, result being shown in (table 3). Comparing our results to those reported by [2] the concentration of sperm of breeding bulls at NLBO, Pokhara was found to be 551.28±41 (10⁵/ml). In comparison to the current findings, Ahmad et al. (2003) found sperm ejaculated from Sahiwal bulls in Pakistan had a lower total concentration 980.86±12.46 (105/ml).

Post-thaw Motility of the Sperm

Jersey bull semen was reported to have an overall post-thaw motility of sperms of 59.13±5.29. Similarly, an overall post-thaw motility of sperms in Holstein semen was found to be 59.24±7.23. There was no significant difference between the species and post-thaw motility of frozen semen (p-value= 0.78) shown in (table 4).

Similarly, post-thaw motility of sperm from three geographical regions i.e. Baglung-Myagdi, Parbat and Nawalpur were found to be 59.34±4.95 %, 57.87±7.18 % and 59.75±5.96 % respectively (p=0.57), result shown in (table 5).

The post-thaw motility of frozen semen was found to be higher in tropical region i.e. 59.75% followed by temperate and sub-tropical region i.e. 59.34% and 57.87% respectively but there was no significant difference between geographical region and post-thaw motility of sperm (p= 0.57).

In comparison to the current findings, [6] reported somewhat lower post-thaw motility in Zebu × taurus bulls (ranging from 51.02% to 55.87%). The current findings are not in line with [7] who showed increased post- thaw motility of sperm from HF bulls (62.50±3.4%) and Czech Fleckvieh bulls (58.67±4.2%). In contrast to the current findings, [8] reported reduced subjective post-thaw motility of sperm in ejaculates of HF bulls (38.31±1.24%). Similarly, the current finding is higher than [9] report of decreased total post-thaw motility of sperms (43.24±0.35%) ejaculated from Sahiwal bull in Pakistan.

Viability of the Sperm

The viability of the sperm for Jersey and Holstein bulls were found to be 14.24 ±3.11% and 12.36±3.71% respectively. The highest viability in the 3rd hour was found to be 14.24% for Jersey and was followed by Holstein i.e. 12.36%, shown in (table 6).

There was significant difference between the species and viability of sperm (p=0.05).

The viability of the sperm in three geographical regions i.e. Baglung-Myagdi, Parbat and Nawalpur were found to be 13.56±4.53 %, 15.00 ± 6.84% and 14.24±3.57% respectively (p=0.70) a represented in (table 7).

The highest viability was seen in subtropical region i.e. 15%, followed by tropical and temperate region i.e. 14.24% and 13.56% respectively. There was no significant difference between viability and geographical region. (p=0.70).

Effect of liquid nitrogen level in Post-thaw Motility of the Sperm

The post thaw motility was 58.00± 3.72, 52.17 ±3.51, 48.75±5.13 & 33.34±3.56 at full, 3/4^th, 1/2^nd and 1/4^th of the straw level of liquid nitrogen respectively for Jersey bulls. Similarly, it was 52.75±8.73, 48.63±3.44, 42.75±4.82 and 29.17±4.38 at full, 3/4^th, 1/2^nd and 1/4^th of the straw level of liquid nitrogen respectively for Holstein bulls, data presented in (table 8). The overall percentage of post thaw motility was significantly higher (P value=5.08E-19) when the level of liquid nitrogen was full and 3/4^th as compared to 1/2^nd and 1/4^th level of straw.

At the full, 3/4^th, 1/2^nd, and 1/4^th levels of liquid nitrogen, respectively, the post-thaw motility was 45-60%, 45-55%, 40- 50%, and 20-40% [10]. In comparison to the 1/2^nd and 1/4^th level of straws, the overall percentage of post-thaw motility was substantially greater (P<0.05) when the liquid nitrogen level was full and 3/4^th of the straw level [11]. In Sahiwal and Red Sindhi bulls, the proportion of post-thaw motility was considerably greater (P<0.05) at the straw level compared to the 1/4^th level of straws. After ten days of storage, the percentage of post-thaw motility in frozen semen straws gradually decreased in a two-liter liquid nitrogen container that did not need to be refilled.

The post-thaw motility of sperms was shown to diminish with a steady reduction in the volume of liquid nitrogen in the container, based on the percentage of motility at different levels. The amount of liquid nitrogen should thus not drop below the halfway point of the straw during storage to promote improved post-thaw motility.

Conclusion

Sperm concentration, post-thaw motility, and sperm viability were the three main areas of focus for the study’s microscopic quality analysis of bull semen. A variety of breeds (Jersey and Holstein,) and geographical areas (Tropical, Sub-tropical, and Temperate) provided data for the collection and analysis. Sperm viability differed considerably among breeds but not regions, while other microscopic quality metrics of bull semen, such sperm concentration and post-thaw motility, did not significantly change based on breed. When it comes to sperm quality, Jersey bulls have consistently outperformed Holstein bulls in terms of concentration and viability.

There was no significant difference in species and concentration of sperm for frozen semen although the highest concentration was found in Jersey semen followed by Holstein. The highest post-thaw motility was observed in Jersey semen followed by Holstein semen. There was no significant difference between the species and post-thaw motility of frozen semen. The highest viability in the 3rd hour was found to be for Jersey. There was significant difference between the species and viability of sperm (p=0.05). Given that the acrosome includes enzymes essential for the penetration of the zona pellucida and the surrounding cellular layers, freezing and thawing procedures have the potential to seriously harm the membrane structures of the sperm head. According to reports, acrosome integrity has a stronger correlation with frozen bull semen fertility than motility. However, sperm motility plays a crucial role in the processes that lead to gamete contact. A decrease in post-thaw sperm velocity and in the percentage of motile cells that are advancing indicates cellular damage that may lessen the likelihood of fertilization. The ½level of liquid nitrogen maintains minimum required post thaw motility in Jersey but in case of Holstein it must be ¾ of tank to ascertain minimum quality. The post thaw motility of sperms decreased on progressive decline in the level of liquid nitrogen in container. Therefore, to ensure better post thaw motility, the level of liquid nitrogen should not fall below 1/2^nd level of straw while storage.

Ethics approval

We author hereby declare you that the article entitled Assessment of the Quality and Performance of Jersey and Holstein Frozen Semen in Gandaki Province, Nepal has no harm to any animals while preparing the manuscript.

Data Availability

Data available in National Livestock Breeding Office, Pokhara were taken into consideration as a reference.

Competing Interest

We declare that an article entitled Assessment of the Quality and Performance of Jersey and Holstein Frozen Semen in Gandaki Province, Nepal has no competing interest among the authors.

Funding

Self partially supported by National Livestock Breeding Office, Pokhara.

Authors Contribution

R Kandel conceived and designed the study, led the research implementation, performed data analysis, and was the primary author of the manuscript. He coordinated all stages of the project and ensured the scientific integrity of the work. S Paudel contributed to field data collection, assisted in semen quality assessment, and supported the interpretation of results. He also participated in manuscript review and editing. K Sapkota and J. Pandaya provided guidance on methodology, assisted in laboratory procedures, and contributed to drafting and reviewing and editing the manuscript for intellectual content. D Tamang supervised the work throughout the research period.

Acknowledgements

• Mahendra Malla, National Livestock Breeding Office, Department of Livestock Services, Pokhara, 33700, Nepal.

• Rupesh Shrestha, Veterinary Hospital and Livestock Service Experts Center, Syangja, 33800, Nepal.

• Rupesh Kandel, PhD, The University of Mississippi, Oxford, MS

• Abinash Bhattarai, National Livestock Breeding Office, Department of Livestock Services, Pokhara, 33700, Nepal

• Suraj Marahatta, Department of Veterinary Medicine and Public Health, Agriculture and Forestry University, Nepal.

• Upasana Ghimire, PhD, Bio nanotechnology & Bio convergence Engineering, Jeonbuk National University, South Korea

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