Objective: A study was carried out to determine the effect of graded levels of moringa leaf meal and season on spermatozoa characteristics of the Pearl Guinea fowl cock in Ghana. Materials and Methods: Thirty-two (32) cocks and one hundred and twenty-eight (128) hens aged one-day-old were used for the study after been reared to attain sexual maturity. A completely randomized design with a 3 x 4 factorial arrangement was used for the experiment. Data collected were analyzed using General Linear Model (GLM) procedure of Statistical Analysis System (SAS for Windows, version 7) and means were separated by the probability of difference (PDIFF) procedure. Results: Result from this study revealed that, the highest sperm motility occurred in birds fed with the control diet (77.12%) while birds fed with 15% Moringa oleifera leaf meal (MOLM) had the lowest sperm motility (55.83%). Semen pH, sperm count and normal spermatozoa increased with increasing dietary moringa leaf meal. Semen pH was higher in birds fed with 15% MOLM (8.00) and lowest in birds fed with the control diet (7.70). Sperm count was higher in birds fed with 15% MOLM (4.63×109 mL) and lowest in birds fed with 9% MOLM (2.33×109 mL). Normal spermatozoa was higher (p<0.05) in birds fed with 15% MOLM (85.42%), whereas, 9% MOLM (74.17%) and the control diet (74.58%) had similar (p>0.05) but significantly lower values for normal sperm. Season had no significant (p>0.05) effect on spermatozoa characteristics except motility and sperm count. Sperm motility was higher (p<0.05) in the dry season (74.09%), whereas minor (67.18%) and major rainy (65.93%) seasons had similar (p>0.05) but significantly lower values for sperm motility. Sperm count was higher (p<0.05) in the major rainy season (4.04×109 mL) and lower in the dry season (1.89×109 mL). There was no significant (p>0.05) effect of moringa leaf meal×season interaction on spermatozoa characteristics. Conclusion: This study concluded that spermatozoa quality and quantity increased with increasing levels of MOLM in the diet of Guinea fowl cocks.
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The production of Guinea fowls plays a major role in bridging the protein gap in developing countries including Ghana where the average daily consumption is far below recommended levels1,2. However, the productivity of Guinea fowl in Ghana has been limited due to higher protein prices, seasonal variation and poor reproductive performance. Seasonal changes can lead to hens not laying at certain times of the year. Short day length affects egg laying because it affects hormonal production and hence reproduction3. Poor nutrition can lead to protein deficiency, which can affect fertility, hatchability and sperm production. Protein sources are especially limiting factors for Guinea fowl and poultry feed production in the tropics4. Hence, there is the need to search for locally available alternative sources of protein for use as poultry feed ingredient. One possible source of cheap protein for Guinea fowl is the leaf meal of some tropical legume and plants5. Recently, there has been interest in the utilization of moringa (Moringa oleifera) commonly called horseradish tree or drumstick tree, as a protein source for livestock6,7. Moringa leaves have quality attributes that make them a potential replacement for soyabean meal or fish meal in non-ruminant diets.
Despite the nutritive and medicinal value of Moringa oleifera, there is scanty information regarding its effect on sperm production in Guinea fowls. Guinea fowls are known to be seasonal breeders with little or no eggs during the dry season3. The study was carried out to determine the effect of dietary moringa leaf meal and season on semen production of the Pearl Guinea fowl in Ghana.
MATERIALS AND METHODS
The study was conducted at the Poultry Unit of the Department of Animal Science Education, University of Education, Winneba, Mampong-Ashanti. Mampong-Ashanti lies in the transitional zone between the Guinea savanna zone of the north and the tropical rain forest of the south of Ghana.
Mampong-Ashanti lies between latitude 07°04’ degree north and longitude 01°24’ degrees west with an altitude of 457 m above sea level. Maximum and minimum annual temperatures recorded during the study period were 30.6 and 21.2°C, respectively8,9. Rainfall pattern in the district is bimodal, occurring from April to July (major rainy season) and again from August to November (minor rainy season), with about 1224 mm per annum. The dry season occurs from December to March10,11.
A total of 160 pearl Guinea fowl pullets were selected from the initial stock obtained from Akate Farms Co. Limited in Kumasi after brooding and used for the study. The birds (32 males and 128 females) were distributed in a completely randomized design in a 3×4 factorial arrangement. The factors that were considered in the experiment included; Factor I: Season at 3 levels (Dry season-December-March, Major rainy season-April-July and Minor rainy season -August-November) and Factor II: Different levels of Moringa leaf meal (MOLM) [Moringa leaf meal at four levels: 0% MOLM (T1, Control), 9% MOLM (T2), 12% MOLM (T3) and 15% MOLM (T4)]. All treatment combinations (Season and different levels of moringa leaf meal combination: 0% MOLM: Dry season, Major rainy season and Minor rainy season, 9% MOLM: Dry season, Major rainy season and Minor rainy season, 12% MOLM: Dry season, Major rainy season and Minor rainy season, 15% MOLM: Dry season, Major rainy season and Minor rainy season) were used. Each treatment was replicated four times and had ten birds per replicate. The birds in each replicate were housed in one pen. One male (four months old) was paired with four Guinea hens.
A total of 16 experimental cages were used for rearing the birds, each measuring 1.4 m×1.34 m and housed 10 birds. The floor was concreted and wood shavings were used as litter for the birds. Removable wooden feeding troughs measuring 0.8 m×0.04 m×0.03 m were used for feeding the growers. In each cage, 4.5 L watering trough was used for supplying water ad libitum for the growers. The experimental diets were supplied to the birds ad libitum throughout the experimental period (Table 1). Vaccination and other routine poultry practices were also carried out. The weather records for the experimental period are shown in Table 2 and 3.
Spermatozoa characteristics: Mature males were trained for eight weeks to be responsive to release off semen. Semen was collected using the dorso-ventral massage method12 from birds randomly selected from each of the replicates. The testes located at the dorsum were stroked and massaged until there was protrusion of the cloacae. The semen was then milked and collected using a rubber pipette and transferred to collection vials. Semen was analyzed immediately after collection at the Science laboratory, College of Agriculture Education, University of Education, Winneba, Mampong-Ashanti.
Physical characteristics of spermatozoa: Sperm motility and debris were assessed by examination of a drop of semen (5 μL) under the microscope at 10× magnification as described by Hutt13 using Olympus BX43-Standard laboratory Microscope manufactured by Mason Technology Company Ltd, India. Sperm motility was determined as the number of cells that were motile in a volume of semen and it was reported as percent cells per millilitre (cells mL1) or millions of cells per millilitre (×106 mL1)14. Semen pH was measured by using a pH meter (specially treated paper blot that changes colour according to the pH of the specimen that it is exposed to)13. Sperm count was determined as the number of spermatozoa in millilitre of semen sample by counting the spermatozoa in a counting chamber and was reported as millions of cells per millilitre (M mL1 or ×106 cells mL1)13.
Morphological characteristics of spermatozoa: Sperm morphology was determined as the measurement of the shape of sperm cells and was reported as percentage normal sperm cells and abnormal spermatozoa cells (big head cells, double tail cells). The percent normal and abnormal spermatozoa were counted after preparing smears and staining them with eosin and nigrosin according to the methods described by López-Rull and Gil14. Semen volume was measured by using calibrated micro pipette, the semen was aspirated and the volume was reported in millilitres (mL) as described by Burrows and Quinn12.
Spermatozoa cell differential characteristics: Maximal leucocyte and erythrocyte concentration were determined as number of epithelia cell, white blood cells and red blood cells in milliliter of semen sample and was reported as millions of cells per milliliter (1×106 cells mL1)14.
Data analysis: Data collected were analyzed using General Linear Model (GLM) procedure of Statistical Analysis System (SAS for Windows, version 7). The means were separated using the probability of difference (PDIFF) procedure of SAS15.
Proximate composition of MOLM: The proximate components of moringa leaf meal contain higher levels of carbohydrates (26.96±1.52), crude protein (28.91±0.21), metabolizable energy (2043.50±55 kcal kg1) and appreciable levels of crude fibre (13.34±0.078), dry matter (89.64±0.45), ether extracts (5.32±0.21), moisture (10.36±0.075), nitrogen free extracts (43.85±0.11) and total ash (7.13±0.04) as shown in Table 4.
Phytochemical properties in moringa leaf meal: A phytochemical analysis was performed to determine the major class of compounds present in moringa leaf meal and the results are shown in Table 5. The results revealed the presence of high levels of chlorogenic acid as compared to all the other parameters. Kaempferol, quercetin and luteolin were observed to be moderate. However, apigenin was observed to be the lowest among all the parameters measured.
Proximate composition of the breeder diet: The proximate composition of the breeder diet is shown in Table 6. The crude protein, crude fibre and metabolizable energy in the breeder diet increased with increasing levels of dietary moringa leaf meal. The control treatment recorded the highest levels of moisture, ether extract and dry matter in the breeder diet while the 15% moringa leaf meal level recorded the lowest moisture content, ether extract and dry matter in the diet. The diet met the nutrient requirement for Guinea fowls as suggested by Okyere et al.2.
Effect of dietary Moringa leaf meal and season on semen physical characteristics: Dietary moringa leaf meal and season had no significant (p>0.05) effect on sperm debris, however, it showed significant influence (p<0.05) on sperm motility, semen pH and sperm count (Table 7). Increasing levels of moringa leaf meal in the diet decreased sperm motility (p<0.05). The highest sperm motility occurred in birds that were fed the control diet while birds fed diets that contained 15% MOLM had the lowest sperm motility. The pH of the semen was higher (p<0.05) for birds fed the diet that contained 15% MOLM (Table 7) which made it relatively alkaline as compared to the other dietary treatments. Sperm concentration was the highest (p<0.001) in birds fed the diet that contained 15% MOLM diet (Table 8). Season of production had influence (p<0.05) on sperm motility and sperm count but not semen pH and sperm debris (p>0.05) (Table 7). Sperm motility was higher (p<0.05) in the dry season whereas, minor and major rainy seasons had similar (p>0.05) but significantly lower values for sperm motility.
Sperm count was higher (p<0.05) in the major rainy season and lower in the dry season. The highest (p<0.05) sperm concentration was recorded in the major rainy season and was followed by minor rainy season with the dry season being the least. Two-way interaction effect of season and dietary moringa leaf meal had little or no influence (p>0.05) on sperm motility, semen pH and sperm count.
Effect of dietary Moringa leaf meal and season on spermatozoa morphological characteristics: Dietary moringa leaf meal had no significant (p>0.05) effect on the number of abnormal spermatozoa and semen volume (Table 8). Dietary moringa leaf meal influenced (p<0.05) the percentage of normal sperm count. Increasing levels of moringa leaf meal in the diet increased the percentage of normal spermatozoa produced. Season had little or no effect (p>0.05) on abnormal spermatozoa, normal spermatozoa forms count and semen volume (Table 8). Therefore, similar means were observed across all seasons for those parameters. The combined effect of season and different levels of moringa leaf meal had little or no influence (p>0.05) on abnormal sperm, normal spermatozoa forms count and semen volume. Guinea fowl cocks fed diets that contained 15% MOLM produced the highest (p = 0.003) normal sperm as compared with other dietary treatments. Guinea fowl cocks fed diets that contained 12% MOLM had higher (p<0.05) normal sperm as compared to 9% MOLM and control which had similar values.