Ind. J. Anim. Agric. Sci., 6(1): 80-87, 2024 80
Optimizing Super Kampung Chicken Semen Quality: Impact of Holding Frequency on Macroscopic and Microscopic Attributes
Yolah Anggriani1, Muhammad Amrullah Pagala2, Fuji Astuty Auza2*, Natsir Sandiah2
1Student of the Faculty of Animal Science, Halu Oleo University
2Faculty of Animal Science, Halu Oleo University
Jl. H.E.A Mokodompit Campus Hijau Bumi Tridharma Anduonohu Kendari, Southeast Sulawesi, 93232 Indonesia
Correspondence email: [email protected] ABSTRACT
The advantages of super Kampung chickens include a faster growth period and robust body resistance to diseases. The success of artificial insemination (AI) in poultry is influenced by various factors, such as the holding technique, storage frequency, spermatozoa concentration, male productivity, inseminator skills, insemination time, and reproductive quality. This study aimed to evaluate the impact of holding frequency on the macroscopic and microscopic quality of chicken semen. The research design employed was a completely randomized design (CRD) consisting of three treatments and five replications. The treatments included P1 (semen storage once per week), P2 (semen storage twice per week), and P3 (semen storage three times per week). Data analysis was conducted using ANOVA, and in case of a significant e ffect, the least significant difference test (LSD) was performed. The results indicated that semen storage did not have a significant effect (P>0.05) on the macroscopic quality (volume, consistency/thickness, semen pH, color, and odor) of semen from super Kampung chickens.
Furthermore, different semen storage frequencies did not significantly impact (P>0.05) spermatozoa movement and motility percentage, but had a substantial effect (P<0.05) on spermatozoa concentration in super Kampung chickens.
Keywords: Collection frequency, semen, spermatozoa, super Kampung chicken.
INTRODUCTION
The population of Indonesia continues to increase every year, leading to a rise in the consumption of animal protein. This trend is attributed to the growing knowledge and awareness among the public regarding the importance of consuming nutritious food. Nutritious food, including meat, is obtained through the consumption of various sources, one of which is the highly favored super Kampung chicken farming (Gallus domesticus sp). According to the Central Statistics Agency in 2022, the population of super Kampung chickens in Indonesia, particularly in Kendari, Southeast Sulawesi, reached 739,047 million individuals.
Super Kampung chickens present a lucrative livestock business due to the increasing demand for meat each year. Their advantages include a faster growth period and good resistance to diseases. However, they also have drawbacks, such as higher feed consumption, requiring a balanced nutrient content in the diet to support rapid growth (Ginting, 2015).
The maintenance of super Kampung chickens can be carried out extensively and semi- intensively with the aim of improving their genetic quality. Genetic improvement can be achieved using various technologies, and one such method is artificial insemination (AI). AI involves the introduction of thawed semen, diluted or liquefied, to enhance the genetic quality
Ind. J. Anim. Agric. Sci.
e-ISSN: 2715-1697
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Ind. J. Anim. Agric. Sci., 6(1): 80-87, 2024 81 of livestock rapidly, producing high-quality offspring in large quantities with human and specialized tool assistance (Darwati, 2019).
The frequency of semen collection significantly influences its quality, affecting the viability and fertility of spermatozoa in the female reproductive tract. Semen quality is assessed both macroscopically and microscopically. Macroscopic semen quality is evaluated based on volume, color, viscosity, and pH, while microscopic semen quality is determined by concentration, mass motility, and motility percentage (Susilawati, 2011).
Excessive collection frequency can lead to a decrease in semen volume, concentration per ejaculation, and a reduction in spermatozoa motility and libido. Irregular and too frequent semen collection prevents the reproductive organ cells from regenerating mature spermatozoa in large quantities, primarily due to the extended processes of spermatozoa formation, maturation, and storage in male avian reproductive organs. If the collection frequency surpasses the quantity of semen filled in the vas deferens, it can negatively impact spermatozoa motility (Mulyadi, 2007).
The success of artificial insemination in poultry is influenced by several factors, including collection technique, storage frequency, spermatozoa concentration, male productivity, inseminator skills, insemination timing, and overall reproductive health (Yuwanta, 2016). The semen collection or harvesting process aims to obtain high-quality semen in both quantity and quality for further processing as liquid or frozen semen for artificial insemination purposes.
MATERIALS AND METHODS Materials
The materials used in this research consist of 15 male super Kampung chickens aged 11- 13 months with body weights ranging from 2,256 to 2,419 kg. The tools utilized in this study include 15 individual cages, feeding dishes, water containers, a set of microscope equipment, glass slides, coverslips, microtubes, pH paper with a scale of 0-14, micropipette, photositometer, 1 mL pipette, and tissue.
Methods
The chickens were individually housed in cages measuring 75 cm in length and 60 cm in width. They were maintained for two weeks before semen collection. Feeding was conducted twice daily at 7:00 AM and 3:00 PM local time, with ad libitum access to water. The provided feed was AD II production from PT. Japfa Comfeed Indonesia, Tbk.
The research design employed a Completely Randomized Design (CRD) with 3 treatments and 5 replications. The treatments included P1 (semen collection once per week), P2 (semen collection twice per week), and P3 (semen collection three times per week).
Data Analysis
The obtained data were tabulated and analyzed using analysis of variance (ANOVA). If there were treatments showing a significant effect, the Duncan test was conducted for further analysis.
RESULTS AND DISCUSSION Super Kampung Chicken Semen Color
Based on Table 1, it is evident that different semen collection frequencies result in 100%
milky white color of super Kampung chicken semen. The research findings indicate that varying semen collection frequencies do not affect the color. This aligns with the results of a
Ind. J. Anim. Agric. Sci., 6(1): 80-87, 2024 82 study by Yunna et al., (2014), reporting that super Kampung chicken semen is milky white.
Putranto et al., (2020) also obtained similar results, indicating that the color and consistency of semen in Burgo chickens are on average milky white and thick. This corresponds with Pratama's statement (2011) that the color and consistency of semen determine sperm concentration. Thick and milky white semen indicates a high sperm concentration, while watery and clear semen suggests a low concentration. The higher the spermatozoa concentration, the higher the semen consistency, and the cloudier the semen color.
Table1. Percentage of Super Kampung Chicken Semen Color
Treatment Semen Color
Milk White (%)
P1 100
P2 100
P3 100
On the other hand, clear-colored super Kampung chicken semen has low concentration and excessive plasma. Ankanegara (2011) states that clear-colored semen leads to lower percentages of fertilized eggs and egg hatching rates. The color of semen generally serves as an indicator of spermatozoa concentration in a single ejaculation. The color represents the normalcy of super Kampung chicken semen. Good-quality super Kampung chicken semen has a milky white color that is not transparent to light. The milky white color is due to the large wavelength bias of spermatozoa, making it appear dark and thick (Putranto et al., 2020). As stated by Yunna et al., (2014), changes in color occur due to disruptions in one of the male chicken's reproductive channels caused by inflammation and injuries, resulting in a change in semen color to gray due to mixing with pus and red due to mixing with blood.
Semen Smell
Based on the documented data in Table 2, it is observed that the odor of super Kampung chicken semen remains consistent as the characteristic odor, even with different semen collection frequencies. This finding indicates that variations in semen collection do not have a significant impact on the odor quality of super Kampung chicken semen.
Table. 2. The Semen Smell of Super Kampung Chicken
Treatment Parameter
Sement Smell
P1 Sperm Special
P2 Sperm Special
P3 Sperm Special
The results are consistent with the viewpoint of Hidayat (2001), who stated that super Kampung chicken semen, under normal conditions, will retain its characteristic odor, namely, the musky scent. This statement also supports previous findings by Suyadi et al., (2012), suggesting that semen under normal conditions generally exhibits a specific characteristic odor.
Consistency (Viscosity) of Semen
Based on Table 3, different semen collection frequencies yield consistent results of thick semen consistency. This indicates that the semen is of good quality and capable of producing a high spermatozoa concentration. The evaluation of semen consistency involves tilting the semen collection tube; if the semen moves quickly, it indicates a watery consistency, whereas slow movement indicates a thick consistency.
Ind. J. Anim. Agric. Sci., 6(1): 80-87, 2024 83 Table 3. Consistency of Semen Super Kampung Chicken
Treatment Parameter
Consistency
P1 Thick
P2 Thick
P3 Thick
This aligns with the statement from Lubis et al., (2012), where the consistency or viscosity degree is assessed by tilting the tube in a well-lit area and then standing it upright after a few moments. If the semen adhering to the tube descends slowly after being stood upright, it indicates a thick consistency. Generally, super Kampung chicken semen has a thick consistency (Hijriyanto et al., 2017). Hijriyanto et al., (2017) state that the color and consistency of semen determine spermatozoa concentration; thick and dark milky white semen indicates a high spermatozoa concentration, whereas watery and clear semen suggests a low concentration.
Semen Volume
The analysis of variance results indicates that the overall mean semen volume is 0.32 mL. This suggests that different semen collection frequencies do not have a significant influence (P>0.05) on semen volume. This finding aligns with previous studies by Wiyanti et al., (2013) and Danang et al., (2012), who also reported an average semen volume of approximately 0.3 mL for super Kampung chickens.
Table 4. Super Kampung Chicken Semen Volume (mL/ejaculate) with Frequency Different Shelters
Replications Treatment
P1 P2 P3
1 0,5 0,33 0,26
2 0,3 0,3 0,33
3 0,2 0,3 0,3
4 0,47 0,33 0,27
5 0,3 0,33 0,28
Average± SD 0,36 ± 0,12 0,32 ± 0,02 0,29 ±0,03 Frequent daily ejaculations can lead to a decrease in semen volume, spermatozoa concentration, and spermatozoa motility. This is attributed to the ejaculation process, where semen is expelled from the storage site in the distal vas deferens, which is adjacent to the cloaca. Whether through natural mating or collection techniques, this process can result in a reduction or emptying of the storage location.
Degree of Acidity (pH) of Semen
The analysis of variance results indicates that different semen collection frequencies do not have a significant impact (P>0.05) on semen pH. The overall mean pH of super Kampung chicken semen is 7.5. This result is consistent with the normal pH range for Kampung chicken semen, which is around 7, as stated by Sopiyana et al., (2006), indicating that local chicken semen typically has a pH of 7.0-7.5. This finding is in close agreement with the study by Zahra (2017), who reported an average semen pH for Kampung chickens ranging from pH 7 to 7.13.
The role of acidity level is crucial as it can affect the viability of spermatozoa. High or low pH values can lead to spermatozoa death (Kartasudjana, 2001).
Ind. J. Anim. Agric. Sci., 6(1): 80-87, 2024 84 Table 5. Degree of Acidity (pH) of Super Kampung Chicken Semen
Replications Treatment
P1 P2 P3
1 7,7 7,6 7,6
2 7,3 7,8 7,4
3 7,3 7,4 7,6
4 8,0 7,4 7,4
5 7,7 7,4 7,2
Average ± SD 7,6 ± 0,3 7,52 ± 0,2 7,44 ± 0,1 The acidity level (pH) was determined by dipping pH paper into the semen collection tube (microtube). The acidity level or pH of semen is influenced by the anaerobic metabolism process of spermatozoa, producing high levels of lactic acid, which can lead to accumulation and consequently affect the increase or decrease in pH values (Yunna et al., 2014).
Evaluation of the Quality of Super Kampung Chicken Spermatozoa
Based on Table 6 it is evident that the percentage of mass movement of super Kampung chicken spermatozoa falls within the normal range, classified as (+++). P1 indicates the highest percentage of mass movement (+++) at 92.9%, P3 has the lowest at 91.74%, and P2 has a percentage of mass movement (+++) at 91.86%. These results are consistent with the findings of Almahdi et al., (2014), who reported a spermatozoa mass movement of 84% for Bangkok chickens. According to Garner and Hafez (2000), the motility in birds typically ranges from 60-80%.
Table 6. Percentage of Super Kampung Chicken Spermatozoa Mass Movement (%)
Replications Treatment
P1 P2 P3
1 91,7 91,4 91,7
2 91,7 92,9 91,6
3 93,3 90,6 91,8
4 93,3 92,3 92,3
5 93,3 92,1 91,3
Average ± SD 92,9 ± 0,8 91,8 ± 0,9 91,3 ± 0,4
Information: P1 (semen collection once a week), P2 (Semen collection 2 times a week), P3 (Semen collection 3 times a week).
Sujoko et al., (2009) state that the criteria for assessing mass movement are 1) +++ (very good, with large, abundant, and thick waves), and 2) ++ (good, with small, thin, infrequent, unclear waves, and slow movement). Mass movement of spermatozoa reflects the movement of individual spermatozoa. This is supported by the opinion of Mardalestari (2005) that mass movement of spermatozoa reflects the movement of individual spermatozoa. The more active and numerous the moving spermatozoa, the better the mass motility (thicker and faster movement). Mass movement correlates with spermatozoa motility and concentration, consistent with the study by Puspitasari (2014), which states that a visible, large, dense, and actively moving wave contains many active motile spermatozoa. Conversely, a thin and slow- moving wave contains fewer active motile spermatozoa. Meanwhile, a mass movement rated as (+) does not show waves but only individual progressive motile movements (Naji, 2006).
Based on the results of the Tukey's Honestly Significant Difference (HSD) test, it is evident that treatment P1 has a significantly higher spermatozoa concentration (P<0.05) compared to treatments P2 and P3. Additionally, treatment P2 has a significantly higher
Ind. J. Anim. Agric. Sci., 6(1): 80-87, 2024 85 spermatozoa concentration than treatment P3. The average spermatozoa concentration in super Kampung chicken semen ranges from 8,312 to 11,477 × 10^6 mL. This result is smaller when compared to Kusumawati's study (2020), which reported a concentration of 13,697 × 10^6 mL for Kampung chickens. Semen concentration is influenced by semen collection frequency, libido, feed, temperature, and season. Ankanegara (2011) stated that the optimal frequency of semen collection results in high concentrations, typically once a week.
Table 7. Super Kampung Chicken Spermatozoa Concentration (×106/mL)
Replications Treatment
P1 P2 P3
1 12.303 9.615 7.554
2 11.648 10.461 8.255
3 11.987 9.755 7.981
4 10.573 10.349 9.121
5 10.874 9.967 8.649
Average ± SD 11.477 ± 2,74c 9.967 ± 3,88b 8.312 ± 3,51a Further analysis reveals that the spermatozoa concentration in super Kampung chickens for each treatment is highest in treatment P1 (11,477 × 10^6 mL) and P2 (9,967 × 10^6 mL) with semen collection once a week and twice a week, respectively. The lowest concentration is observed in treatment P3 (8,312 × 10^6 mL) with semen collection three times a week.
Collecting semen too frequently in a short period can decrease spermatozoa concentration. This finding differs from the study by Hijrianto et al., (2017), which reported that semen collection frequency in Bangkok chickens (1 time, 2 times, and 3 times a week) did not affect the resulting concentration. This difference may be attributed to variations in chicken breeds, age, husbandry practices, feed provided, and the number of samples used.
The spermatozoa concentration produced falls within the normal range for poultry, which is 2.5 × 10^9 to 3.5 × 10^9 mL of semen (Iskandar et al., 1998). Putranto et al., (2020) added that differences in chicken spermatozoa concentration are influenced by factors such as feed, body weight, and chicken breed. Larger chicken breeds generally have higher spermatozoa concentrations.
Percentage of Individual Motility of Super Kampung Chicken Spermatozoa
The analysis of variance results indicates that different semen collection frequencies do not have a significant effect (P>0.05) on the percentage of spermatozoa motility in super Kampung chickens. Based on Table 4.8, the average percentage of motility in super native chicken semen ranges from 91.74% to 92.9%.
Table 8. Average Percentage of Motility (%) of Super Kampung Chicken Semen
Replications Treatment
P1 P2 P3
1 91,7 91,4 91,7
2 91,7 92,9 91,6
3 93,3 90,6 91,8
4 93,3 92,3 92,3
5 93,3 92,1 91,3
Average ± SD 92,9 ± 0,8 91,8 ± 0,9 91,3 ± 0,4
Ind. J. Anim. Agric. Sci., 6(1): 80-87, 2024 86 The highest motility average is 92.9% for treatment P1, followed by P2 with an average motility of 91.86%, and the lowest is P3 with an average motility of 91.74%. These results are relatively consistent with the study by Hijrianto et al., (2017), where semen collection once a week resulted in 90.3% motility, twice a week resulted in 90.5%, and three times a week resulted in 91.1%.
Semen collection once and twice a week is considered infrequent. The difference in results may be due to the fact that P1 has a one-day interval between semen collections, potentially allowing for the maturation of spermatozoa in the vas deferens, while P3 has a one- day interval between semen collections. Another possible cause is external factors such as body weight, feed consumption, and heat stress due to uneven weather conditions. Semen collection three times a week is considered optimal. Initially, semen collection three times a week showed a low percentage. This is because the chickens were not accustomed to semen collection. As time progressed, the chickens became accustomed to semen collection, allowing for the maturation of spermatozoa in large quantities.
CONCLUSION
Based on the research findings, it was observed that the more frequently semen is collected, the concentration of spermatozoa tends to decrease. The treatment showing the highest concentration of spermatozoa is in treatment P1, and this difference is statistically significant (P<0.05) when compared to treatments P2 and P3. Semen collection once a week in treatment P1 resulted in a higher concentration of spermatozoa compared to collections twice a week and three times a week in treatments P2 and P3, respectively.
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