Cock Semen Research Articles

Potential Use of Coconut Water and Saline Solutions as Diluents for Guinea Cock Semen during Short-term Storage at Ambient Temperature

Potential Use of Coconut Water and Saline Solutions as Diluents for Guinea Cock Semen during Short-term Storage at Ambient Temperature

Dietary supplementation of spirulina and canthaxanthin boosts laying performance, lipid profile in blood and egg yolk, hatchability, and semen quality of chickens.

This study examined the impacts of dietary spirulina (SP) and canthaxanthin (CX) supplementation to laying hens on reproductive performance and cock's semen quality evaluation. Three hundred and sixty of indigenous Sinai chickens (300 hens + 60 cocks), 48-weeks-old, were randomly divided into five treatments; a control and four supplemented (0.6 g SP mg/kg, 0.8 g SP mg/kg, 6 mg CX/kg diet, and 8 mg CX/kg diet) with six replications of 10 females and 1 male per treatment for laying trial and 1 male for semen evaluation. The animals were fed with experimental diets for 12 weeks. Supplementing 6 mg CX/kg diet led to an increase in the final body weight and egg production traits, while 8 mg CX/kg diet resulted in an improving feed conversion ratio. Supplementation of 0.6 g SP mg/kg or 6.0 mg CX/kg diet resulted in the highest egg production. The heaviest egg weight and egg mass were induced by 0.8 g SP mg/kg. Eggs laid from hens fed 0.6 g SP/kg diet had a higher concentration of cholesterol than those fed 0.8 g SP/kg and those fed 6.0 and 8.0 mg CX/kg. Cocks fed 6 mg CX/kg and those fed 0.8 g SP/kg diet had the greatest sperm concentration. The supplementation of 0.8 g SP mg/kg diet or both 6 and 8 mg CX/kg diet enhanced hatchability. So, dietary spirulina (0.8 mg/kg) and canthaxanthin (6 mg/kg) supplementation to Sinai laying hens and cocks could be used to improve productive and reproductive and performance.

Optimizing liquid storage duration of two poultry species semen with plant based extender

Liquid storage of semen preservation for poultry species is deemed to be the only reliable and cost effective method compare to cryopreservation. This experiment was conducted to examine the preservation capacity of Tris coconut-water orange juice extender (TCWOE) on tom and cock semen for artificial insemination. TCWOE was prepared. Semen ejaculates were collected and pooled from five toms and cocks separately. The pooled semen from each species was divided into two portions and in one portion of each species the extender was added in the ratio of 1:3 (semen: extender), making a total of four treatments. Experimental design used was a complete randomized design consisting of three trials. Semen microscopic parameters like motility, viability, membrane integrity, and acrosome integrity were examined and recorded for freshly extended semen and preserved semen at 4-8oC every 12 h interval till 72 h. The motility and membrane integrity of freshly extended cock semen was significantly higher (P<0.05) compared to motility of extended tom semen and membrane integrity of un-extended tom semen. During storage up to 24 h, extended tom and cock semen showed significantly higher (P<0.05) motility and membrane integrity compare to un-extended semen. Extended cock semen had significant (P<0.05) higher percentage motility, livability and membrane integrity compared to un-extended and extended tom semen from 36 h to 72 h of storage. However, no significant difference (P>0.05) was observed for acrosome integrity. Conclusively, cock semen survived longer and had better results considerable for artificial insemination than tom semen up to 72 h storage. However, the extender was beneficial for storage of both species semen.

Spermatozoa Survival Rate in Stored Cock Semen Extended With Diluent Fortified With Natural Antioxidant Sources under Room Temperature

An experiment was conducted in a completely randomized design with five treatments in triplicates. Four extenders were formulated: Lake Extender; Coconut water-Orange juice Extender (CWOE), Egg yolk-Tomato juice Extender (EYTE) and Milk based-Carrot Extender (MCE), were used to extend cock semen constituting treatments 2, 3, 4 and 5 respectively, while the control (Treatment 1) was unextended semen. Semen was harvested from 8 proven Isa breeder cocks aged 35-40 weeks, pooled and evaluated. The ejaculate was divided into 5 and each part was extended with the extenders at ratio 1:2 (Semen: Extender). The extended and un-extended semen samples were kept under room temperatures (26.6 27.4°C) with relative humidity of 64.0-74.0% for in vitro assessment at 2 hrs interval until percentage motility was less than 60%. The results showed that the mass activity, sperm motility and percentage livability reduced p<0.05) as the time of storage increased. Mass activities of unextended semen and the one extended with coconut water-orange juice were significantly (P<0.05) higher than that of other treatments at 0 hour period of storage. Sperm motility in treatments 1 (95.0%), 3 (93.3%) and 4 (88.3%) was not significantly (p>0.05) different from one another, but they was significantly (p<0.05) higher than treatments 2 (40.0%) and 5 (71.7%). At 2 hours of storage, the mass activity and percentage sperm motility in treatment 2 were significantly (P<0.05) lower than other treatments, while live sperm cells were significantly lower for Treatment 3 than other treatments. At 4 hours of storage, mass activity and motility were zero in Treatment 2, while Treatment 1 recorded the highest values (66.7% and 93.5% respectively). Percent motility in Treatments 3, 4 and 5 were 55.0, 50.0 and 43.3%, respectively. However, percent live sperm cells were significantly (P<0.05) higher in treatments 2, 4, 5 and 1 than the in treatment 3. Both CWOE and EYTE possessed a good keeping quality for semen storage. The extenders fortified with natural antioxidants can be used for on-farm insemination in poultry within two hours of collection.

Spermatozoa Survival Rate in Stored Cock Semen Extended With Diluent Fortified With Natural Antioxidant Sources under Room Temperature

An experiment was conducted in a completely randomized design with five treatments in triplicates. Four extenders were formulated: Lake Extender; Coconut water-Orange juice Extender (CWOE); Egg yolk-Tomato juice Extender (EYTE) and Milk based-Carrot juice Extender (MCE), were used to extend cock semen constituting treatments 2, 3, 4 and 5 respectively, while the control (Treatment 1) was unextended semen. Semen was harvested from 8 proven Isa breeder cocks aged 35-40 weeks, pooled and evaluated. The ejaculate was divided into 5 and each part was extended with the extenders at ratio 1:2 (Semen: Extender). The extended and un-extended semen samples were kept under room temperatures (26.6-27.4oC) with relative humidity of 64.0-74.0% for in vitro assessment at 2 hrs interval until percentage motility was less than 60%. The results showed that the mass activity, sperm motility and percentage livability reduced p<0.05) as the time of storage increased. Mass activities of unextended semen and the one extended with coconut water-orange juice were significantly (P<0.05) higher than that of other treatments at 0 hour period of storage. Sperm motility in treatments 1 (95.0%), 3 (93.3%) and 4 (88.3%) was not significantly (p>0.05) different from one another, but they was significantly (p<0.05) higher than treatments 2 (40.0%) and 5 (71.7%). At 2 hours of storage, the mass activity and percentage sperm motility in treatment 2 were significantly (P<0.05) lower than other treatments, while live sperm cells were significantly lower for Treatment 3 than other treatments. At 4 hours of storage, mass activity and motility were zero in Treatment 2, while Treatment 1 recorded the highest values (66.7% and 93.5% respectively). Percent motility in Treatments 3, 4 and 5 were 55.0, 50.0 and 43.3%, respectively. However, percent live sperm cells were significantly (P<0.05) higher in treatments 2, 4, 5 and 1 than In treatment 3. Both CWOE and EYTE possessed a good keeping quality for semen storage. The extenders fortified with natural antioxidants can be used for on-farm insemination in poultry within two hours of collection.

Reproductive organ weight of Nigerian indegenous cocks fed diet with graded levels of tumeric (Curcuma longa)

A study was conducted to evaluate the reproductive performance/reproductive organ weight of Nigerian indigenous chicken (cock) fed diet with graded levels of turmeric rhizome powder (TRP). The objectives of the study were to determine the effect of TRP on the characteristics (relative organ weight), and to evaluate the effect of TRP supplemented diet on the reproductive organ weight of Nigerian Indigenous cocks. One hundred and twenty adult local breeding cocks divided into four groups with 30 birds in each group were used for the study. Each group was further replicated into three replicates of 10 birds per replicate. The birds in the experimental groups designated as T1 , T2 , T3 , and T4 were fed with TRP supplemented diet at various inclusions, 0.0%, 0.25%, 0.50 %, and 0.75% respectively for eight weeks. The following parameters were measured, full reproductive organ (g), right vas deferens (g), paired vas deferens (g), right epididymis (g), left epididymis (g), paired epididymis (g), right testis (g), left testis (g) and paired testis. The result showed a gradual (p < 0.05) decline in reproductive morphometric weight and were significantly (p < 0.05) improved in the TRP supplemented group better than the control group. In conclusion, Turmeric powder supplemented diet at 0.25, and 0.5 % inclusions performed better in the reproductive morphometric of Nigerian indigenous cocks, although care should be exercised in prolonged supplementation above 0.5 % inclusion to prevent its deleterious effect on the organ weight. The reproductive organ weight of Nigerian indigenous cocks fed with Turmeric powder supplemented diet as observed in this study compared favorably and even better than the control T1. This is an indication that Turmeric powder has a significant propensity to enhance the reproductive performance of our indigenous cock semen since semen quality is determined to a large extent by the size of the reproductive organ. Although, the Turmeric powder has no significant deleterious effect on the major reproductive organs which make it very safe for inclusion in the diet of our local indigenous cocks. Also from the study, to a better performance, good morphometric quality and size of the Nigerian indigenous cocks and with no negative effect on the reproductive performance of Nigerian Indigenous Cocks, 0.5% inclusion level is highly recommended.

EFFICIENCY OF USING A COMBINATION OF MONO- AND DISAC-CHARIDES IN A DILUENT FOR FREEZING ROOSTER SEMEN

РЕФЕРАТ НА АНГЛИЙСКОМDifferent combination of saccharides can provide better semen protection during freezing/thawing cycle. Until now, the disaccharide maltose has not been used as a component of the medium for cryopreservation roosters’ semen. Since maltose is not involved in carbohydrate metabolism of spermatozoa, there is an assumption about its role in strengthening the structure of the glycocalyx, which is a progressive evolutionary cellular structure that regulates specific cellular adaptations to a certain temperature, chemical and other paratypical effects. In this work, in order to increase the fertility of frozen/thawed semen, we tested a combination of saccharides in the diluent for cryopreservation of roosters’ semen. For the first time, maltose has been proven to be effective in combination with fructose in a diluent to increase the fertility of frozen/thawed roosters’ semen. The aim of the study was to determine the optimal concentration of the test component of maltose based on the diluent of the Leningrad Cryoprotective Medium (LCM) (1984) for freezing the semen of roosters and determining the time of maintaining the functional usefulness of frozen/thawed cock semen in the genital tract of the hen. The experiment was carried out in the Center for Collective Use Genetic collection of rare and endangered chicken breeds (the Russian Research Institute of Farm Animal Genetics and Breeding, 2020) on a breed of Russian white chickens (♂n = 10, ♀n = 30) at the age of 46-50 weeks. Three variants of media for cryopreservation of roosters’ semen with different ratios of saccharides were evaluated. In each group, the Mal-10 (fructose 0.72 %, maltose 0.166 %), Mal-20 (fructose 0.64 %, maltose 0.326 %) and LCS-control (fructose 0.8 %, maltose 0 %), there were 10 hens for insemination. The results show not only an increase in the total percentage of fertilized eggs when using frozen/thawed semen in the Mal-10 (92.6 %) and Mal-20 (86.3 %) groups compared to the LKS-control group (74.7 %), but also an increase in the duration of the functional usefulness of spermatozoa in the genital tract of hen within 5 days at the level of native sperm. Counting the points of interaction of spermatozoa with the perivitelline membrane of the yolk 5 days after the last insemination showed that the functional ability of spermatozoa is much higher when using experimental media containing maltose, 67.0 pcs/cm2 for Mal-10 and 110.7 pcs/cm2 for Mal-20 vs. 40.1 pcs/cm2 in maltose-free LKS-control. The longest duration of the functional usefulness of spermatozoa was noted in the Mal-20 group. Even on day 15 after the last insemination, the fertilization capacity of frozen/thawed semen was recorded at the level of 20 %. A regression equation was drawn up for the relationship of egg fertilization with the points of interaction of spermatozoa with the perivitelline membrane of the yolk. To obtain an egg fertilization level of ≥ 80 %, the functional usefulness of frozen-thawed spermatozoa (expressed through the number of points of interaction with the perivitelline membrane of the yolk) should be ≥ 60 pcs/cm2. This was achieved by using an experimental diluent with 0.326 % maltose. The results obtained open up the possibility of using cryopreserved semen not only in preserving the gene pool, but also in breeding programs.

ЭФФЕКТИВНОСТЬ ИСПОЛЬЗОВАНИЯ КОМБИНАЦИЙ САХАРИДОВ В СРЕДАХ ДЛЯ КРИОКОНСЕРВАЦИИ СПЕРМЫ ПЕТУХОВ

Different combination of saccharides can provide better semen protection during freezing/thawing cycle. Until now, the disaccharide maltose has not been used as a component of the medium for cryopreservation roosters’ semen. Since maltose is not involved in carbohydrate metabolism of spermatozoa, there is an assumption about its role in strengthening the structure of the glycocalyx, which is a progressive evolutionary cellular structure that regulates specific cellular adaptations to a certain temperature, chemical and other paratypical effects. In this work, in order to increase the fertility of frozen/thawed semen, we tested a combination of saccharides in the diluent for cryopreservation of roosters’ semen. For the first time, maltose has been proven to be effective in combination with fructose in a diluent to increase the fertility of frozen/thawed roosters’ semen. The aim of the study was to determine the optimal concentration of the test component of maltose based on the diluent of the Leningrad Cryoprotective Medium (LCM) (1984) for freezing the semen of roosters and determining the time of maintaining the functional usefulness of frozen/thawed cock semen in the genital tract of the hen. The experiment was carried out in the Center for Collective Use Genetic collection of rare and endangered chicken breeds (the Russian Research Institute of Farm Animal Genetics and Breeding, 2020) on a breed of Russian white chickens (♂n = 10, ♀n = 30) at the age of 46-50 weeks. Three variants of media for cryopreservation of roosters’ semen with different ratios of saccharides were evaluated. In each group, the Mal-10 (fructose 0.72 %, maltose 0.166 %), Mal-20 (fructose 0.64 %, maltose 0.326 %) and LCS-control (fructose 0.8 %, maltose 0 %), there were 10 hens for insemination. The results show not only an increase in the total percentage of fertilized eggs when using frozen/thawed semen in the Mal-10 (92.6 %) and Mal-20 (86.3 %) groups compared to the LKS-control group (74.7 %), but also an increase in the duration of the functional usefulness of spermatozoa in the genital tract of hen within 5 days at the level of native sperm. Counting the points of interaction of spermatozoa with the perivitelline membrane of the yolk 5 days after the last insemination showed that the functional ability of spermatozoa is much higher when using experimental media containing maltose, 67.0 pcs/cm2 for Mal-10 and 110.7 pcs/cm2 for Mal-20 vs. 40.1 pcs/cm2 in maltose-free LKS-control. The longest duration of the functional usefulness of spermatozoa was noted in the Mal-20 group. Even on day 15 after the last insemination, the fertilization capacity of frozen/thawed semen was recorded at the level of 20 %. A regression equation was drawn up for the relationship of egg fertilization with the points of interaction of spermatozoa with the perivitelline membrane of the yolk. To obtain an egg fertilization level of ≥ 80 %, the functional usefulness of frozen-thawed spermatozoa (expressed through the number of points of interaction with the perivitelline membrane of the yolk) should be ≥ 60 pcs/cm2. This was achieved by using an experimental diluent with 0.326 % maltose. The results obtained open up the possibility of using cryopreserved semen not only in preserving the gene pool, but also in breeding programs.

10.4314/sajas.v40i5.65348

This study evaluated the effect of breed on the survivability and motility rate of cryopreserved cock semen. Semen from three cock breeds; White Leghorn (WL), Ovambo (OV) and Potchefstroom Koekoek (PK) was collected by means of the abdominal massage technique. Following semen collection, sperm were analyzed for motility and survivability with the use of contrast light BHTU microscope (20 x magnification). The semen was diluted (1 : 2 v/v) with egg yolk citrate (EYC) (extender A) and thereafter with extender B (EYC + 5% DMSO). The equilibration after each dilution was 2 h at 5 °C. The diluted samples were evaluated for sperm concentration, motility, survivability and pH. The samples were then loaded into straws and cooled in programmable freezer from 5 °C to -20 °C at the rate of 1 °C/minute. Semen straws were then exposed to liquid nitrogen vapour (-80 °C) for five minutes, plunged directly into liquid nitrogen (-196 °C) and stored for a week or more. Frozen straws were thawed at 5 °C and evaluated at 0, 30, 60 and 90 min post-thaw. From the results there was no significant effect of breed on the survival and motility of fresh-diluted and frozen-thawed semen at 30 and 90 min post-thaw in all breeds. The sperm survivability of the PK breed was significantly higher than that of the WL breed. However, there was no sperm survivability difference between PK and OV breed immediately after thawing. The cryopreservation and thawing processes affected the survivability and motility of sperm of all poultry breeds negatively.

The experimental transmission of reticuloendotheliosis virus by cock semen.

Following artificial insemination, the egg-laying rate of a large-scale breeder chicken flock declined by10-15 %. Real-time quantitative polymerase chain reaction (qPCR) analysis detected the presence of reticuloendotheliosis virus (REV) in semen from the breeder cocks used. Six REV strains were successfully isolated from semen randomly extracted from those cocks. Additionally, the whole sequence of SDAUR-S1 was sequenced and analysed. Cock models with continuous production of REV-positive semen were established by intravenous injection with SDAUR-S1. Eggs were then collected from hens after artificial insemination with REV-positive semen, for virus detection. The positive REV antibody rate for egg albumen was 58.3 % and the REV-positive rate for hatched embryos was 8.3 %, which suggested not only that REV can infect cock semen, but can also infect the offspring. In conclusion, the present study is the first to report on the isolation, genome analysis and transmission of REV in cock semen.

Response of Gramapriya hybrid cock spermatozoa to hypo-osmotic swelling test and its correlation with seminal attributes

A total of 114 semen ejaculates were obtained twice a week from 38 weeks old Gramapriya hybrid cocks were studied for Hypo- osmotic swelling test (HOST) to assess membrane integrity and other physical characteristics during the period of 4 months from November, 2008 to February, 2009. The mean value of physical attributes; ejaculate volume (ml), colour/appearance score, pH, mass motility (%), individual motility (%), sperm concentration (X 109/ml), total spermatozoa count (X 109/ejaculate), live spermatozoa count (%) and abnormal sperm count (%) averaged to 0.39±0.02, 3.98±0.03, 7.42±0.01, 3.95±0.03, 84.01±0.04, 2.49±0.03, 0.98±0.04, 91.01±0.29 and 6.58±0.22, respectively followed by mean value of HOST reactive spermatozoa was 92.98±0.26 percent. A significant negative correlation of HOST reactive spermatozoa was observed with abnormal spermatozoa and ejaculate volume whereas; it correlated positively with sperm concentration and initial motility. The study elicited application of HOST to evaluate the Gramapriya hybrid cock semen by characteristic features of twisting and swelling of head of the spermatozoa along with the normal coiling of tail in

The characterisation and cryopreservation of Venda chicken semen

Abstract Objective To characterize Venda cocks semen, find a suitable short-term diluted semen storage temperature, find a suitable cryopreservation cryoprotectant and to investigate cryoprotectant toxicity. Methods Semen was collected from six Venda cocks and evaluated macroscopically for semen volume, pH and sperm concentration. Microscopic sperm characteristics examined included were total motility (rapid, medium and slow) progressive and non-progressive motility. Velocity characteristics included curvilinear and straight-line velocity, average path velocity, linearity, straightness, wobble, amplitude of lateral head displacement and beat cross frequency. Results Results showed that the average semen volume was 0.3 ± 0.1 mL, the pH 6.9 ± 0.4 and the sperm concentration (6.8 ± 79.8) × 109/mL. A positive correlation was observed between body weight and semen volume (r = 0.38). Similarly a significant difference between the initial sperm total motility (TM%) of 87.5 ± 8.6 and samples stored for 24 h at 5 °C (55.0 ± 8.0) and 25 °C (30.6 ± 6.1) was recorded. The percentage live and normal sperm was 87.0% and 93.5% (P Conclusions In conclusion, the Venda cock semen was subsequently found to have a higher TM% when stored in vitro at 5 °C. DMSO and EG were found to be suitable for the cryopreservation of Venda cock semen.

41 THE EFFECT OF AMBIENT TEMPERATURE ON SPERM MOTILITY DURING LIQUID STORAGE OF VENDA COCK SEMEN AND INDIVIDUAL DIFFERENCES IN SPERM CRYOTOLERANCE

Improving techniques for liquid storage of cock semen can increase the efficiency of AI programs in the poultry industry. The aims of the present study were (1) to compare storage of cock sperm for 24 h at 5 and 25°C and (2) to test the cryotolerance of sperm cell motility in individual Venda cocks. Semen was collected with the abdominal massage method, from 6 indigenous Venda cocks. Cocks were 26 weeks of age and were kept under the same conditions. After macroscopic analysis, semen was pooled and diluted (1:2) with Kobidil+ extender and divided into 3 equal parts. Part 1 was evaluated immediately (0 h), part 2 was stored at 5°C and part 3 was stored at 25°C and evaluated for sperm motility and velocity parameters at 4, 8, 12 and 24 h of storage. For cryopreservation, semen was diluted (1:2) with modified Kobidil+ extender supplemented with 8% of dimethyl sulfoxide. Individual ejaculates were equilibrated at 5°C for 4 h and then loaded into the programmable freezer. Then, semen straws were thawed at 5°C. Sperm motility and velocity parameters were evaluated using the Sperm Class Analyzer® system. Six replicates were done per trial. Data were analysed using the statistical programme GenStat®. Treatment means were separated using Fisher's protected t-test least significant difference (P < 0.05). Total sperm cell motility rate was 87.5% and decreased significantly during in vitro storage and was <31% after 24 h at 25°C. Semen samples stored at 5°C showed a total sperm cell motility rate of above 50% after 24 h. There was a slight linear decrease in the percentage of sperm with progressive motility and rapid velocity as the storage period increased, irrespective of the storage temperature. The rapid and medium motility percentages were higher in fresh semen and significantly decreased (P < 0.05) during the incubation period. There was variation in the total sperm cell motility of fresh and frozen semen among cocks. There was no significant difference in variation in non-progressive and medium percentage (P > 0.05) motility in diluted fresh or frozen sperm cells or in the percentage of sperm with rapid motility in thawed semen. There was variation in <25% of the cocks in total sperm motility rate. In summary, cryopreservation reduced sperm cell motility and velocity rates in all the cock semen donors. We found that cryotolerance of cock sperm does vary among males. Furthermore, the lower temperature 5°C was suitable for semen storage of Venda cocks. This temperature (5°C) could potentially improve methods of semen equilibration before cryopreservation. The study was supported by an Agricultural Research Council Parliamentary Grant, Department of Agriculture Forestry and Fisheries and National Research Foundation-GUN No RT21 and 24000 (NRF). The Germplasm Conservation & Reproduction Biotechnologies (GCRB) group is thanked for their support.

Generation of antiviral transgenic chicken using spermatogonial stem cell transfected in vivo

This study was conducted in order to generate anti-viral transgenic chickens through transfected spermatogonial stem cell with fusion gene EGFP-MMx. After injecting fusion gene EGFP-MMx into testes, tissues frozen section, polymerase chain reaction (PCR) and dot blot of testes was performed at 30, 40, 50, 60, 70 and 80 days. Few normal hens were inseminated artificially with cock semen of 60 days. Finally, PCR, reverse transcription-PCR and Western blot were used to detect the integration of exogenous gene EGFP-MMx of the offspring. Dot blot positive rate of testes genome was 72.2%. PCR and RT-PCR positive rate was 25 and 10.53% (4/38) in sperm and F1 blood genome, respectively, while Western blot positive rate of F1 blood was 7.89% (3/38). Taken together, the possibility of cultivating anti-viral transgenic chickens by the method of testes injection which could integrate foreign genes into the genome was proved. Key words : EGFP-MMx fusion protein, spermatogonial stem cell, transgenic chickens.

Generation of antiviral transgenic chicken using spermatogonial stem cell transfected in vivo

This study was conducted in order to generate anti-viral transgenic chickens through transfected spermatogonial stem cell with fusion gene EGFP-MMx. After injecting fusion gene EGFP-MMx into testes, tissues frozen section, polymerase chain reaction (PCR) and dot blot of testes was performed at 30, 40, 50, 60, 70 and 80 days. Few normal hens were inseminated artificially with cock semen of 60 days. Finally, PCR, reverse transcription-PCR and Western blot were used to detect the integration of exogenous gene EGFP-MMx of the offspring. Dot blot positive rate of testes genome was 72.2%. PCR and RT-PCR positive rate was 25 and 10.53% (4/38) in sperm and F1 blood genome, respectively, while Western blot positive rate of F1 blood was 7.89% (3/38). Taken together, the possibility of cultivating anti-viral transgenic chickens by the method of testes injection which could integrate foreign genes into the genome was proved. Key words : EGFP-MMx fusion protein, spermatogonial stem cell, transgenic chickens.

Relationships between cock semen viability and the fertility of artificially inseminated South African indigenous chicken breeds

One hundred and sixty hens and 32 cocks of four different South African indigenous chicken breeds (Naked Neck (NN), Ovambo (OVB), Potchefstroom Koekoek (PK) and Venda (VD) were used in this study. Reproductive performance tests as determined by the number of ejaculations per five minutes of abdominal sexual massage (5ASM) were used to select 16 high performing (HP) and 16 low performing (LP) cocks from a population of 80 cocks. Cocks with≥2 ejaculates/60 min or ≤1 ejaculate/60 min following ASM were considered as HP or LP cocks, respectively. Semen was collected from each cock following 5ASM, evaluated for semen viability and 0.05 mL diluted semen used to inseminate five hens per breed, in each experimental group. Significant differences in ejaculation rates and semen quality and quantity were recorded in the four breeds of cocks - with the HP cocks of the PK breed producing the highest ejaculation rate (4.1 ± 0.1/60 min), ejaculate volume (0.7 ± 0.02 mL), sperm motility (92 Class Test 2 ± 0.7), percentage live sperm (88.4 ± 0.7%) and total sperm per ejaculate. Significant differences were also recorded in hen fertility and subsequent hatchability of set eggs. Semen from the HP cocks were superior to those from the LP cocks, as evidenced in most of the fertility and hatchability parameters, with PK layers producing eggs that gave the highest average egg weight (58.04 ± 0.9 g), fertility (74.3 ± 0.2%), hatchability of set eggs (84.03 ± 0.7%), percentage of normal chicks (98.1 ± 0.7%) and chick weight (37.9 ± 0.3 g). Results suggest that selection of high performing cocks following five minutes of sexual massage prior to semen collection enhances semen quality and quantity leading to significant improvement in the fertility of artificially inseminated hens.

The effect of breed on the survivability and motility rate of

This study evaluated the effect of breed on the survivability and motility rate of cryopreserved cock semen. Semen from three cock breeds; White Leghorn (WL), Ovambo (OV) and Potchefstroom Koekoek (PK) was collected by means of the abdominal massage technique. Following semen collection, sperm were analyzed for motility and survivability with the use of contrast light BHTU microscope (20 x magnification). The semen was diluted (1 : 2 v/v) with egg yolk citrate (EYC) (extender A) and thereafter with extender B (EYC + 5% DMSO). The equilibration after each dilution was 2 h at 5 °C. The diluted samples were evaluated for sperm concentration, motility, survivability and pH. The samples were then loaded into straws and cooled in programmable freezer from 5 °C to -20 °C at the rate of 1 °C/minute. Semen straws were then exposed to liquid nitrogen vapour (-80 °C) for five minutes, plunged directly into liquid nitrogen (-196 °C) and stored for a week or more. Frozen straws were thawed at 5 °C and evaluated at 0, 30, 60 and 90 min post-thaw. From the results there was no significant effect of breed on the survival and motility of fresh-diluted and frozen-thawed semen at 30 and 90 min post-thaw in all breeds. The sperm survivability of the PK breed was significantly higher than that of the WL breed. However, there was no sperm survivability difference between PK and OV breed immediately after thawing. The cryopreservation and thawing processes affected the survivability and motility of sperm of all poultry breeds negatively.

Incubation and Fertility Research Group: {WPSA Working Group 6 (Reproduction)} 2010 Meeting - Tours, France, 23 August 2010

The IFRG meeting 2010 was held in the beautiful city of Tours in France. The topics now reported included the storage and early embryo development; new developments in incubation temperature; and the influence of glucose application to poultry embryos. Two aspects of cryopreservation were considered: primordial germ cells, and in vivo cryopreserved cock semen. The latter was tested to assess its feasibility for use in commercial production or preservation of genetic stocks. With regard to commercial poultry production, the data recorded in one study supported the hypothesis that fertility would be higher and would persist for longer in pens where the birds were able to find a physical refuge from other birds. Other factors affecting the production of turkeys, ducks, pheasants, ostriches, chickens and ducks were discussed, thus fulfilling the meeting's objective of reporting research into the incubation and fertility of all avian species.

Studies on cock semen. II. Effects of frequent ejaculation and breed on some biochemical characteristics

Studies on cock semen. II. Effects of frequent ejaculation and breed on some biochemical characteristics

Studies on cock semen. I. Effects of frequent ejaculation and breed on physical characteristics

Studies on cock semen. I. Effects of frequent ejaculation and breed on physical characteristics