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SEMEN COLLECTION, EVALUATION, ARTIFICIAL INSEMINATION and CORRELATION of SEMEN PARAMETERS with PREGNANCIES in ALPACAS a Capsto

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SEMEN COLLECTION, EVALUATION, ARTIFICIAL INSEMINATION and CORRELATION of SEMEN PARAMETERS with PREGNANCIES in ALPACAS a Capsto SEMEN COLLECTION, EVALUATION, ARTIFICIAL INSEMINATION AND CORRELATION OF SEMEN PARAMETERS WITH PREGNANCIES IN ALPACAS A Capstone Experience Manuscript Presented by Petra Szymkowicz Completion Date: May 2012 Approved By: Stephen R. Purdy, DVM, Veterinary and Animal Science Department, Chairperson Carlos Gradil, DVM, Veterinary and Animal Science Department, Committee Member ABSTRACT Title: “Semen Collection, Evaluation, Artificial Insemination and Correlation of Semen Parameters with Pregnancies in Alpacas” Author: Petra Szymkowicz, Animal Science CE Type: Independent Capstone Thesis Approved By: Dr. Stephen R. Purdy, Veterinary and Animal Science Department Approved By: Dr. Carlos Gradil, Veterinary and Animal Science Department The purposes of this study are to present information regarding semen characteristics and the potential for artificial insemination in 20 male alpacas and examine the correlation for these parameters with pregnancy rates. The samples were collected from an artificial vagina mounted in a breeding phantom and also from the cranial vagina of bred females immediately post copulation. 12 of the 20 males were evaluated 4 or more times using the breeding phantom and 24 postcoital breeding samples were evaluated. The postcoital samples were correlated with pregnancy results in the females in the study as determined by transrectal ultrasound. Male reproductive organs were initially examined by palpation of the testes, and ultrasound of the testes, bulbourethral glands, and prostate gland. Measurements and ultrasound pictures were recorded. Semen parameters evaluated for the samples included volume (phantom breedings only), sperm activity immediately after semen collection, live sperm percentage and sperm morphology from stained slides, and estimated sperm concentration immediately after semen collection (estimated as low, medium, or high). Data is presented on all measured parameters along with pictures. Ability to collect males using the breeding dummy was inconsistent. Of 101 attempted mounts of the phantom, 38% produced collections with a volume greater than 0.1 mL with sperm seen on the slides. Of the 45 collections in which activity percentages were recorded, 33% of these collections were successful with activity percentages greater than 0. A wide variation in semen parameters was found in individual males and among the males collected making the artificial insemination an unlikely prospect in alpacas. Sperm activity, live percentage, normal morphology percentage, and estimated concentration were evaluated with respect to pregnancies achieved in the study females. No direct correlation for any of these parameters was found in these males and females. INTRODUCTION The purpose of this research was twofold. The first was to use a breeding phantom to collect, analyze, and determine normal semen parameters for alpacas as well as to investigate the feasibility of artificial insemination in that species. The second part of this research was conducted using postcoital semen samples to determine the semen parameters necessary to achieve pregnancies in an alpaca. To date there has been very little research conducted that investigates the characteristics of alpaca semen. Prior research performed by Vaughan in 2003, Morton in 2008, Wiggin in 2009, and Mainini in 2010 has proven that collection of alpaca semen is possible with an artificial vagina placed inside a breeding phantom. The problem is that all of these studies have had varying success rates between different males; also the majority of these studies have been conducted with a small sample size. The benefit of being able to collect males using a breeding phantom and artificial vagina is that semen of breeding males can be collected and evaluated prior to breeding to determine their reproductive soundness and to identify any potential fertility problems. Additionally if these males could consistently be collected there is potential for extension of the semen and using it for artificial insemination. Artificial insemination (AI) in other domestic livestock has been successfully used to propagate and preserve superior genetics and increase the availability of superior genetics to breeders. For the alpaca industry this could potentially improve fiber, fertility, conformation, and disease resistance in offspring. Additionally AI would eliminate the need for owners to transport males or females for matings. This would reduce the risk of spreading infectious disease, decrease stress to the animals, and increase farm biosecurity. 1 The problem with AI in alpacas to date is that males have proven difficult to consistently collect. This might be due to the dribbling and viscous nature of the ejaculate but the exact reason it is unknown. This study aimed to determine if a reliable and consistent technique of semen collection is even possible. There is no research correlating semen parameters with achieved pregnancies in alpacas. If specific parameters necessary for conception could be determined then males could be evaluated for potential fertility problems before owners of males and females invested money into breedings. This would effectively increase conception rates and decrease the risk of using a stud that is subfertile. MATERIALS AND METHODS Animals In this study twenty adult Huacaya male alpacas were used (Table 1). They ranged from 2.5 years to 11 years in age with a mean age of 5.4 years. Nine of the twenty males (45%) had experience with natural breeding before; the other 11 (55%) had no prior breeding experience at all. All of the males were borrowed from farms in Western Massachusetts and transported to the UMass Hadley Farm in Hadley, Massachusetts with the exception of one that was owned by the University of Massachusetts and already resided at the farm. All of the males were used for semen collection and evaluation using the breeding phantom and an artificial vagina. Additionally, males B, D, F, K, L, M, N, and P were used in natural service for correlating semen parameters from post‐breeding collections with achieved pregnancies. The males were kept at the University of Massachusetts Amherst, Hadley farm in small groups and were fed ad libitum second cut hay, fresh water, and a specially formulated mineral mix for alpacas. Season permitting, they also had access to ad libitum 2 grass pasture and pelleted grain when necessary to maintain normal body condition. They were housed in separate but adjacent pastures to the females being used for the semen parameter to pregnancy correlation study. These same females were occasionally used to stimulate the libido of the males to persuade them to mount the breeding phantom. The study females (Table 2) were housed under the same conditions as the males. They ranged from 3 to 11 years of age and 0 to 9 previous crias. The females were used for natural service breedings and collecting postcoital semen samples, which were then evaluated with respect whether or not a pregnancy was achieved. The females were also used as teasers for the males to increase libido during semen collections using the phantom. Physical Examination Prior to Breeding Upon arrival at the farm all of the body condition scores of the borrowed males were assessed, and fecal samples were taken from each animal and analyzed for intestinal parasites. Every male was assessed to be in good health and adequate body condition at the start of the study. Prior to breeding all males’ reproductive organs were examined using a 7.5MHz linear ultrasound probe and their testes were measured using calipers. The ultrasound was used to visualize the size and echotexture of the left and right bulbourethral glands, the prostate gland, and both testes. Testicle length and width were measured using calipers and carefully rotating the testicle horizontally in the scrotum. (Figure 1) Table 1 shows the general information about the males used in the trial along with the caliper measurements of their testicles. The average size for the left testicle was 4.2 cm long by 2.8 cm wide with a range of 3.8 cm to 5.0 cm long by 2.2 to 3.8 cm wide. The 3 average for the right testicle was 4.5 cm long by 2.9 cm wide with a range of 3.4 cm to 4.8 cm long by 2.4 to 4.0 cm wide. Figure 2 shows the homogenous appearance expected when viewing a normal testicular parenchyma using ultrasound. Figure 3 shows the ultrasound image of the testicular parenchyma when testicular cysts are present. All males in this study with the exception of Male T had normal homogenous appearing testicular parenchyma in both testes. Male T had hypoechoic regions in both left and right testes, which are consistent with testicular cysts. Figure 1: Testicular measurement (length) using calipers (Pindar and Purdy) 4 Table 1: Study Males Age Left Testicle Right Testicle (length x Natural Breeding Male (years) (length x width) cm width) cm Experience A 6 ‐ ‐ yes B 4 3.9 x 2.2 3.4 x 2.7 yes C 3 4.2 x 2.6 4.1 x 2.4 no D 8 5.0 x 2.8 4.8 x 2.7 no E 6 3.9 x 2.1 3.8 x 2.9 no F 6 4.0 x 3.1 4.1 x 3.1 yes G 6 3.8 x 3.0 3.9 x 2.8 no H 5 4.1 x 2.9 3.9 x 2.7 no I 5 4.1 x 2.9 4.6 x 3.9 no J 5 4.0 x 3.1 4.6 x 3.9 no K 8 4.7 x 2.6 4.6 x 2.6 yes L 4 4.5 x 2.7 5.3 x 3.1 no M 3.5 3.8 x 2.3 3.6 x 2.2 yes N 3 4.4 x 2.4 4.5 x 2.3 no O 11 3.9 x 2.4 3.9 x 2.5 yes P 7 4.6 x 3.1 4.6 x 3.0 yes Q 2.5 4.1 x 3.3 4.8 x 3.2 no R 3 4.0 x 2.8 3.8 x 2.4 no S 6 4.8 x 3.8 4.8 x 4 yes T 5 4.2 x 3.4 4.5 x 3.2 yes Average 5.4 4.2 x 2.8 4.5 x 2.9 ‐ Range 2.5‐11 3.8‐5.0 x 2.2‐3.8 3.4‐4.8 x 2.4‐4.0 ‐ Table 2: Study Females Age Female Number of successful pregnancies in past (years) a 2.5 0 b 3 1 c 5 2 d 7 1 e 5 1 f 13 3 g 14 7 h 11 9 5 Figure 2: Normal testicular ultrasound appearance Figure 3: Abnormal testicular ultrasound appearance consistent with testicular cysts Alpaca Collection Phantom Previous researchers (Wiggin and Purdy, 2009) constructed the breeding phantom from a foam deer archery target, foam padding, and a tanned alpaca hide cover.
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