Diyala Journal of Agricultural Sciences (DJAS), 10(2): 1-11, 2018 Khalil Khalil 1 http://www.agriculmag.uodiyala.edu.iq/ EFFCT OF AUTUMN AND WINTER SEASONS ON SEMINAL TRAITS AND TESTICULAR MEASUREMENTS OF AWASSI RAM LAMBS Raaed I. Khalil ddrr..rraaeedd@@aaggrriiccuullttuurree..uuooddiiyyaallaa..eedduu..iiqq Dept. of Animal Production, College of Agric., Univ. of Diyala, Iraq. ABSTRACT The objective of this study was to compare between the effect of autumn (decrease daylight) and winter (increase daylight) seasons on some reproductive traits in Awassi ram lambs for selecting them on the onset of sexual activity as in their flock . This study was performed at the Ruminant Research Station, Abu Ghraib- belong to the Directorate of Agricultural Researches during the two periods, autumn (17-24 November ) and winter (26 January –3 February) seasons using twelve Awassi ram lambs aged 12-13 month. The study has observed non-significant difference between seasons for semen parameters, seminal plasma parameters, testosterone, cortisol concentrations, testicular measurements and reaction time. However, this study revealed positive correlation between ejaculate volume and total spermatozoa per ejaculate(P≤0.01), mass activity and individual motility (P≤0.01), semen protein with seminal triglyceride (P≤0.01), seminal protein and seminal cholesterol (P≤0.05) and seminal cholesterol with seminal Aspartate Aminotransferase (AST) (P≤0.05).In conclusion, the costs were reduced depending on autumn season for estimation the sexual activity of ram lambs to select them for the sheep flocks. Key Words: Autumn, winter, Awassi ram lambs, seminal traits. INTRODUCTION Many important factors have impact on reproductive capacity and semen production of rams such us, body weight, photoperiod, climatic temperature and management. These factors can reflect huge variations of semen characteristics (Karagiannidis et al., 2000; Al-Mjamaii, 2011; Mukasa-Mugerwa and Ezaz, 1992; Perez et al., 1997; Ibrahim et al., 2016) as well as they decline metabolic impulses at hypothalamic-pituitary axis (Barb and Kraeling, 2004). The sexual activity of a ram reaches its peak at the fall and this activity drops down of the late winter of spring and summer. Decreasing time lengths stimulates the secretion of plasma SSH, ICSH and testosterone in rams, while increasing day light inhibits these hormones (Hafez and Hafez, 2000). "Lambs born in autumn from long hours of daylight, then to short hours of daylight Recievied: 30/4/2017 Accepted: 14/6/2017 http://www.agriculmag.uodiyala.edu.iq/ http://www.agriculmag.uodiyala.edu.iq/ mailto:dr.raed@agriculture.uodiyala.edu.iq Diyala Journal of Agricultural Sciences (DJAS), 10(2): 1-11, 2018 Khalil Khalil 2 http://www.agriculmag.uodiyala.edu.iq/ accelerates this sexual development, depending on the breed" (Hafez and Hafez, 2000). The sexual capacity of rams can be measured by semen quality, hormones concentration, in addition to the testicular measurements, whose correlated with testicular development (Daudu,1984; Al-Rubaay et al., 2011; Moghaddam et al., 2012 ; Ibrahim et al., 2016). Therefore, there are difference in testicular size testosterone level, spermatogenesis efficiency and semen quality between breeding and non-breeding season in ram (Kafi et al. 2004; Moghaddam et al. 2012). In addition, the serum testosterone rises at an earlier age in the ram lamb (Chakraborty et al. 1989). In Iraq, Awassi sheep and other local breed are continues breeding animal (Alwahab et al., 1982), but the first limiting factor for sexual activity was associated with nutrition and availability of the pasture in late spring and summer (May and Jun) when daylight increasing, where the ewes mated. The objective of the current first study in the literature of Iraq was to determine effect of autumn (decrease daylight) and winter (increase daylight) seasons on some sexual capacity characteristics of Awassi ram lambs according to short of daylight. MATERIALS AND METHODS This study was undertaken at the Ruminant Research Station-Abu Ghraib which belongs to the Directorate of Agricultural Researches during the two periods; autumn (17-24 November) and winter (26 January –3 February) using twelve Awassi ram lambs (aged 12-13 month). Semen was collected using artificial vagina (Evans And Maxwell, 1987), after false mount for increasing libido (Fahey et al., 2012). Immediate examination includes: ejaculate volume by graduated tube, colour, mass activity, individual motility (Walton, 1933), dead spermatozoa percentage (Swanson and Bearden, 1951), abnormal spermatozoa percentage (Hancock, 1951), spermatozoa concentration (Salisbury et al., 1943), spermatozoa per ejaculated according for the following equation: Spermatozoa per ejaculate =Vol. Eja.(ml) × Sperm. concentration. In addition of pH using special Indicator paper (Ural Yt-U, Germany). Chemical analysis of seminal plasma includes: triglyceride measurement according the method described by Buccolo and David (1973) by using Kit (Biolabo Company, France), total protein measurement depending on the method described by Green et al., (1982) by using Kit manufactured by Syrbio http://www.agriculmag.uodiyala.edu.iq/ http://www.agriculmag.uodiyala.edu.iq/ Diyala Journal of Agricultural Sciences (DJAS), 10(2): 1-11, 2018 Khalil Khalil 3 http://www.agriculmag.uodiyala.edu.iq/ Company, Syria, glucose measurement depending on the method described by Cooper (1973) using Kit manufactured by Biomaghreb Company, Spanish, cholesterol examination used the method described by Allain (1974) using Kit produced by Biomaghreb Company, Spanish, ALT and AST examination depending on the method described by Reitman and Frankel (1957) using Kit produced by Syrbio Company, Syria. To elucidated the testicular development, the testis measurements (Scrotal circumference, Right testes length, left testes length, Right testes diameter and Left testes diameter). Concentration of serum testosterone and cortisol hormone were determined using ELISA method (Testosterone and cortisol : Monobind Inc., USA). The sensitivity of the testosterone assay was 0.38pg and the intra- and inter assay coefficients of variation was 5.67% and 6.1% respectively. The sensitivity of the cortisol assay was 62.5pg, the intra-assay 6.9% and inter-assay 8%. Statistical analysis The data analyzed using according to Completely Randomized Design (CRD), with two groups and 12 replicates, and the differences between means were tested using least significant differences at 0.05. RESULTS AND DISCUSSTION Table 1 noticed non-significant effect between autumn and winter in ejaculated volume (ml), semen color, mass activity (%), individual motility (%), dead spermatozoa (%), abnormal spermatozoa (%), spermatozoa concentration /ml ×109, total spermatozoa per ejaculate ×109 and semen pH, although increase both the ejaculated volume (0.89 ml), mass activity (70.83%), individual motility (75.00 %) and retrogress in life spermatozoa (dead spermatozoa 10 %) in autumn season as compared with Winter season. The increase in dead spermatozoa in autumn may returned to the increasing of pH and mass activity. The relationship between mass activity and both pH and dead spermatozoa was positive. In addition, the correlation between pH and dead spermatozoa was positive (Table 5). http://www.agriculmag.uodiyala.edu.iq/ http://www.agriculmag.uodiyala.edu.iq/ Diyala Journal of Agricultural Sciences (DJAS), 10(2): 1-11, 2018 Khalil Khalil 4 http://www.agriculmag.uodiyala.edu.iq/ Table 1.Effect of autumn and winter seasons on semen parameters for Awassi ram lambs Significance Season Indicator Winter Autumn SE X SE X N.S 0.09 0.76 0.06 0.89 Ejaculated volume (ml) N.S 0.00 3.00 0.00 3.00 Semen color N.S 0.97 67.50 2.74 70.83 Mass activity % N.S 0.90 71.25 1.95 75.00 Individual motility % N.S 0.86 4.00 1.63 10.00 Dead spermatozoa % N.S 0.53 1.21 0.62 2.00 abnormal spermatozoa% N.S 0.30 4.16 0.42 3.53 Spermatozoa concentration (ml ×109) N.S 0.49 3.15 0.44 3.16 Total spermatozoa per ejaculate (×109) N.S 0.02 6.82 0.06 6.88 Semen pH N.S: No significant. Tables (2 and 3) revealed non-significant effect between autumn and winter in seminal triglycerides (mg/dl), seminal protein (g/dl), seminal glucose (mg/dl), seminal cholesterol (mg/dl), seminal ALT (IU/L), seminal AST (IU/L), testosterone (ng/mmol) and cortisol concentration (mg/dl). However, the testosterone hormone was increased, while cortisol hormone, ALT and AST were decreased in autumn season . Table 2.Effect of autumn and winter seasons on some seminal plasma parameters for Awassi ram lambs Significance Season Indicator Winter Autumn SE X SE X N.S 7.10 74.17 7.03 65.67 Seminal triglycerides (mg/dl) N.S 1.91 22.03 2.46 20.13 Seminal protein (g/dl) N.S 0.54 1.92 0.59 2.33 Seminal glucose (mg/dl) N.S 2.57 28.08 2.92 32.50 Seminal cholesterol (mg/dl) N.S 0.73 7.18 0.72 5.78 Seminal ALT (IU/L) N.S 42.32 275.00 31.35 271.31 Seminal AST (IU/L) N.S 0.22 0.74 0.17 0.86 Testosterone hormone (ng/mmol) N.S 0.20 1.47 0.14 1.34 Cortisol hormone (mg/dl) N.S: No significant. http://www.agriculmag.uodiyala.edu.iq/ http://www.agriculmag.uodiyala.edu.iq/ Diyala Journal of Agricultural Sciences (DJAS), 10(2): 1-11, 2018 Khalil Khalil 5 http://www.agriculmag.uodiyala.edu.iq/ Table 3. Effect of autumn and winter seasons on serum testosterone and cortisol hormone for Awassi ram lambs Significance Season Hormones Winter Autumn SE X SE X N.S 0.22 0.74 0.17 0.86 Testosterone hormone (ng/mmol) N.S 0.20 1.47 0.14 1.34 Cortisol hormone (mg/dl) N.S: No significant. Table 4 displays non-significant effect between autumn and winter seasons in scrotal circumference (cm), right testes length (cm), left testes length (cm), Right testes diameter (cm), Left testes diameter (cm) and Reaction Time (sec.), with preference autumn season in diameter of right and left testes in addition of reaction time as compared with winter season. Table 4. Effect of autumn and winter seasons on testicular measurements and reaction time in for Awassi ram lambs Significance Season Indicator Winter Autumn SE X SE X N.S 0.64 29.17 0.85 28.46 Scrotal circumference (cm) N.S 0.33 14.00 0.26 13.38 Right testes length (cm) N.S 0.28 13.71 0.26 13.25 Left testes length (cm) N.S 1.47 68.21 1.85 70.33 Right testes diameter (cm) N.S 1.19 67.54 1.44 69.88 Left testes diameter (cm) N.S 3.64 27.52 3.50 22.59 Reaction time (sec.) N.S: No significant. In this study, the positive correlation (P≤0.01) between ejaculated volume with total spermatozoa per ejaculate, mass activity with individual motility (P≤0.01) and dead spermatozoa (P≤0.05) was found (Table 5). The relationship was positive between: seminal protein and seminal triglyceride (P≤0.01), seminal protein with seminal cholesterol (P≤0.05) as well as seminal cholesterol and seminal AST (Table 6). http://www.agriculmag.uodiyala.edu.iq/ http://www.agriculmag.uodiyala.edu.iq/ Diyala Journal of Agricultural Sciences (DJAS), 10(2): 1-11, 2018 Khalil Khalil 6 http://www.agriculmag.uodiyala.edu.iq/ http://www.agriculmag.uodiyala.edu.iq/ http://www.agriculmag.uodiyala.edu.iq/ Diyala Journal of Agricultural Sciences (DJAS), 10(2): 1-11, 2018 Khalil Khalil 7 http://www.agriculmag.uodiyala.edu.iq/ http://www.agriculmag.uodiyala.edu.iq/ http://www.agriculmag.uodiyala.edu.iq/ Diyala Journal of Agricultural Sciences (DJAS), 10(2): 1-11, 2018 Khalil Khalil 8 http://www.agriculmag.uodiyala.edu.iq/ Ali and Taha (2012) found that better sexual activity of Awassi rams (adult ram) in Iraq during autumn season, especially in October. Moghaddam et al., (2012) observed the best quality of semen in Iranian crossbreed rams (adult ram) during autumn season. The current results didn’t exhibite the significance effect obviously between all parameters and measurements in spite of some exceptions of these parameters such, increase of ejaculated volume, mass activity, Individual motility and testosterone concentration while cortisol concentration, ALT and AST decrease in autumn as compared with winter season. These parameters is very important as indicators for sexual activity, as well as was the best coefficient of variation in some parameters in autumn season (Table 7), Thus, absences the significant effects (as its progress young rams to the maturity) and coefficient of variance that’s may be give a good indicator on sexual behavior of Awassi ram lambs (Although continues breeding animal) in short daylight may be led to select rams of the onset sexual` activity as good rams of flock. Table 7. Coefficient of variation between autumn and winter seasons in some Parameters for Awassi ram lambs Season Indicator Winter Autumn C.V C.V 41.63 21.63 Ejaculated volume 4.99 13.4 Mass activity 4.36 8.99 Individual motility 74.81 56.52 Dead sperms 153.27 107.13 Abnormal sperms 402.24 245.44 Sperm concentration 187.56 205.73 Total sperms 218.32 186.39 Reaction Time 99.30 147.40 Testosterone Hormone 211.27 281.95 Cortisol Hormone REFERENCES Ali, S. H. and M. B. Taha. 2012. Effect of reproductive season and semen collection on the concentration of some reproductive hormones of Awassi rams. Iraqi J. of Vet. Sci. 26(2): 75-81. Allain, C. C., L. S. Poon, C. S. G. Chon, W. Richmond and P. C. Fu. 1974. Enzymatic determination of total serum cholesterol. Clin. Chem., 20:254- 265. http://www.agriculmag.uodiyala.edu.iq/ http://www.agriculmag.uodiyala.edu.iq/ Diyala Journal of Agricultural Sciences (DJAS), 10(2): 1-11, 2018 Khalil Khalil 9 http://www.agriculmag.uodiyala.edu.iq/ Al-Mjamaii, R. I. K. 2011. Study the effect of Awassi ewe lambs body weigh on it's puberty and the changes in some biochemical and hormonal parameters an it's productivity. Ph. Dissertation. College of Agriculture and Forestry. University of Mosul. Al-Rubaay, H. M., M. K. Al-Dolaymi and N. N. Al-Anbari. 2011. Effect of Seasonal variation in testes and epididymis of Awassi rams. J. Kerbala University /Scientific. 9(2):213-219. Alwahab, R. M. H., W. K. Al-Murani and J. E. Alkass. 1982. Some observations on the breeding season of the Awassi breed in Iraq. Wld. Rev. Anim. Prod. (18):58-63. Barb, C. R. and R. R Kraeling. 2004. Role of leptin in the regulation of gonadotropin secretion in farm animals. Anim. Reprod. Sci. 82: 155–167. Buccolo, G. and H. David. 1973. Quantitative determination of serum triglycerides by the use of enzymes. Clin. Chem. 19(5):476-482. Chakraborty, P. K., L. D. Stuart and J. L. 1989. Puberty in the male Nubian goat: Serum concentrations of LH, FSH and testosterone from birth through puberty and semen characteristics at sexual maturity. Anim. Reprod. Sci. 20: 91. Cooper, G. R. 1973. Methods for determining the amount of glucose in blood. Crit. Rev. Clin. Lab. Sci., 4:101-145. Daudu, C. S. 1984. Spermatozoa output: testicular sperm reserve and epididymal storage capacity of the Red Sokoto goats indigenous to Northern Nigeria. Theriogenology. (21): 317-324. Evans, G. and C. Maxwell. 1987. Salamon’s artificial insemination of sheep and Goats. Sydney: Butterworth. pp. 66-76, 158-161. Fahey, A. G., P. Duffy and S. Fair. 2012. Effect of exposing rams to a female stimulus before semen collection on ram libido and semen quality. J. Anim. Sci. 90:3451–3456. Green, S. A., S. J. Jenkins and P. A. Clark. 1982. A comparison of chemical and electrophoretic methods of serum protein determination in clinically normal domestic animals of various ages. Cornell Vet. 72: 416-426. Hafez, B. and E. S. E Hafez. 2000. Reproduction in Farm Animal.7th ed. Lippincott Wilkins, Philadelphia. Hancock, J. L. 1951. A staining technique for the study of temperature shock in semen. Nature. Land. (167):323-324. http://www.agriculmag.uodiyala.edu.iq/ http://www.agriculmag.uodiyala.edu.iq/ Diyala Journal of Agricultural Sciences (DJAS), 10(2): 1-11, 2018 Khalil Khalil 11 http://www.agriculmag.uodiyala.edu.iq/ Ibrahim, A. G., R. I. Khalil and A. A. Alani. 2016. Study of the effect of body weight on some reproductive characteristics in Awassi rams. Diyala Agric. Sci. J. 8(2):6-15. Juma, K. H. and J. E. Alkass. 1996. Awassi sheep in Iraq. Dirasat Agric. Sci. 23 (3): 200-207. Kafi, M., M. Safdarian and M. Hashemi. 2004. Seasonal variation in semen characteristics, scrotal circumference and libido of Persian Karakul rams. Small. Rumin. Res. 53: 133–139. Karagiannidis, A., S. Varsakeli, C. Alexopoulos and I. Amarantidis. 2000. Seasonal variation in semen characteristics of Chios and Friesian rams in Greece. Small Rumin. Res. 37:125–130. Moghaddam, G. H., M. M. Pourseif and S. A. Rafat. 2012. Seasonal variation in semen quantity and quality traits of Iranian Crossbred rams. Slovak J. Anim. Sci. 45(3): 67-75. Mukasa-Mugerwa, E. and Z. Ezaz. 1992. Relationship of testicular growth and size to age, body weight and onset of puberty in Menz ram lambs. Theriogenology. 38: 979–988. Perez, C., A. Lopez, A. Castrillejo, D. B. Laborde,T. Gastel, R. Tagle, D. Queirolo, J. Franco, M. Forsberg, H. Rodriguez-Martinez. 1997. Reproductive seasonality of Corriedale rams under extensive rearing conditions. Acta Vet. Scand. 38: 109–117. Reitman, S. and S. Frankel. 1957. Colometric methods for the determination of serum glutamic oxaloacetic and glutamic pyruvic transaminase. Ames .J. Clin. Path., 20: 56-63. Salisbury, G. W., G. H. Beek, I. Elliet and E. L. Willett. 1943. Rapid method of estimating the number of spermatozoa in bull semen. J. Dairy Sci. 26: 483- 486. SPSS Inc. 2011. Statistical Package for Social Science version 20 for windows LEAD Technologies. Inc. USA. Swanson, E. W. and H. J. Beardon. 1951. An eosin nigrosin stain differentiating live and dead bovine spermatozoa. J. Anim. Sci. 10: 981-987. Walton, A. 1933. Technique of artificial insemination. Mp. Bur. Anim. Genet. 56, Iiius- Edinburgh. http://www.agriculmag.uodiyala.edu.iq/ http://www.agriculmag.uodiyala.edu.iq/ Diyala Journal of Agricultural Sciences (DJAS), 10(2): 1-11, 2018 Khalil Khalil 11 http://www.agriculmag.uodiyala.edu.iq/ المنوي ومقاييس الخصية للحمالن تأثير فصلي الخريف والشتاء في بعض صفات السائل العواسية الذكرية رائد إبراهيم خليل ddrr..rraaeedd@@aaggrriiccuullttuurree..uuooddiiyyaallaa..eedduu..iiqq ، العراق.جامعة ديالى ،كلية الزراعة ،قسم اإلنتاج الحيواني المستخلص تهدف الدراسة الحالية للمقارنة بين فصلي الخريف )انخفاض ساعات النهار( والشتاء )ارتفاع ساعات النهار( على بعض الصفات التناسلية لدى الحمالن العواسية الذكرية في بداية نشاطها التناسلي ة/ دائرة البحوث قسم بحوث الثروة الحيواني فيأُجريت اذه الدراسة لقطعان األغنام. ا تختياراا ككورال شباط( 3 –كانون أول 26والشتاء ) (تشرين الثاني 24-17لموسمي الخريف ) الزراعية/ وزارة الزراعة . بينت نتائج الدراسة عدم وجود فروق معنوية في اشهر 13-12باستخدام حمالن عواسية ككرية بعمر الكورتيزول وبعض مقاييس صفات المني وصفات البالزما المنوية وتركيز ارموني التستستيرون و عالي المعنوية موجب أظهرت الدراسة وجود معامل ارتباطووقت تحفيز الذكور. فضال عن الخصية (P≤0.01) ،الحركة الجماعية والحركة الفردية، بين كال من حجم القذفة وعدد الحيامن في القذفة الواحدة ومعنوي وجود معامل ارتباط موجب عنفضال البروتينات والداون الثالثية في البالزما المنوية (P≤0.05) كلسترول مع وال البروتينات وكلسترول البالزما المنوية بين كال منAST في البالزما فادة من فصل الخريف للتقييم المبكر للحمالن الذكرية إمكانية اإلاالستنتاج من اذه الدراسة او المنوية. م وبالتالي تقليل كلف اإلنتاج. واتختياراا كذكور جيدة لقطعان األغنا صفات مني. ية،حمالن عواسية ككر، الشتاء ،الخريف :الكلمات المفتاحية http://www.agriculmag.uodiyala.edu.iq/ http://www.agriculmag.uodiyala.edu.iq/ mailto:dr.raed@agriculture.uodiyala.edu.iq mailto:dr.raed@agriculture.uodiyala.edu.iq