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VETS90096 Veterinary Professional Practice

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  • Course Code: VETS90096
  • University: The University Of Melbourne
  • Country: Australia

Question:

Development of a Practical Method of Predicting the Presence of Long Spermatids in Alpacas.

Inroads have been made into the development of more advanced breeding technologies in camelids around the world (Vaughan, 2002, Abraham, 2015). The aim of this research is to act as an adjunctive and practical method for the early selection of the best quality sperm particularly in alpacas.

The urogenital tract including testicular anatomy in camelids has been described by others (Brown, 2000, Sumar, 1996).

The aims of this study are to add to an emerging body of work contributing to prediction/selection of the suitability to breed in male alpacas. To attempt to further equip farmers with practical advice in choosing the best sires they can. This will assist in narrowing in on the best genetic gains for alpacas in Australia by choosing for desirable traits for fleece diameter and colour and also males with higher fertility potential. Due to the relatively slow genetic gain in this species, sire choice is paramount. In both cattle and sheep, testicular size has direct correlation to the volume of sperm the animal can produce. It is then reasonable to expect that camelids are similarly correlated. 80 pairs of alpaca testicles were measured in this study. According to results found by D. B. Galloway in 2000 that total scrotal width was the best predictor for the amount of spermatozoa present. He recommended that male alpacas can then be ranked according to the onset of testicle development.   

Solidification of an easy and repeatable method of testicular cell maturation prediction is important to advance the alpaca fibre industry in Australia and further afield. The 80 pairs of testicles were measured in situ and then assessed histologically to aid in defining the measurable characteristics of highly fertile potential sires.

Traditionally sires have been selected on the basis of early prepuce-penis liberation as yearlings. Only 8% of males have achieved this at age 1. Sumar states that sires are also chosen by their testicular size based on an assumption that there would be a direct relationship as seen in other species (Sumar 1996)

A Swedish study found that as a single measure, testicular length was a good predictor of the presence of sperm (Abraham et al 2016).

Materials and methods:

Castration of 80 animals took place on a property at Merrijig in North-East Victoria, Australia.

Before surgical castration:

The ear tag number, birth date, weight, thoracic circumference, whithers height and body condition score was recorded for each animal. It was ensured that both testicles were present and then the length and width of both the right and left testicles were measured – both length and width using digital calipers.  

Castration:

Anaesthesia and analgesia – buccalgesic designed to have oral xylazine added (10mg/ml oral meloxicam) with oral xylazine (20mg/ml) added for sedation. Overall concentration once mixed were 8mg/ml of meloxicam and 4mg/ml of oral xylazine.

Standing surgical castration, consisting of two scrotal incisions parallel to and 1cm lateral to the median raphe. Incision length was sufficient to remove the testicle and to allow for good post-operative drainage. Closed castration method, removing the adipose from the wound to minimise the risk of infection or delayed healing. An angiotribe was applied for one minute prior to testicle removal, to crush the cord. This method was used to guard against the risk of pulling the testicular artery off the aorta. If it was deemed that the oral analgesia was not effective enough for some individuals, 1ml lignocaine was injected into each spermatic cord.

Post surgical care:

Trisolfen was sprayed onto/into the surgical incisions post castration.

All castrated animals were turned out onto clean, dry, good quality pasture. Alpacas were walked around for five minutes twice a day to keep wounds open and draining well. Walking the animals helped in detection of reluctance to move and these animals could be re-examined. Animals were observed at 24 and 48 hours post surgery, then again at 10 and 24 days post surgery by Dr Jane Vaughan.

Sample collection and preparation:

Testicles were immediately stored in Bouin’s fixative. A sharp scalpel blade was used to take a single 5mm longitudinal section from the left and right testicles. All samples were then stored in Bouin’s fluid for 24 hours, so to preserve the cellular nuclear integrity within the delicate tissue. Sections were then transferred to 70% alcohol before histological preparation.

Microtome slices were then prepared with Haematoxylin and Eosin stain.

Each microscope slide was then examined for the stage of spermatogenesis in the seminiferous tubules. 40 seminiferous tubule cross-sections were examined per testicle slice, one slice per testicle.

Results:

The data was split into two age groups, animals below three years and those equal to 3 years and above (see table 1 for details).

Table 1:

Age group

(years)

n

Mean age

(years ± sem*)

Range (years)

Mean weight

(kg ± sem)

Mean testicular length

(cm ± sem)

Percent of males with spermatids

Mean percent (± sem) of tubules with spermatids

Range

< 3

45

2.4 ± 0.03

2.0-2.9

46.2 ± 1.09

3.57 ± 0.07

86%

51.4% ± 4.59

0-89%

≥ 3

35

3.5 ± 0.07

3.0-4.5

 

59.7 ± 1.32

3.98 ± 0.06

94%

75.4% ± 3.53

0-94%

 

Every alpaca > 3.2 years (n=25) had long spermatids present. Within the older group of alpacas, 80% of animals have over 70% long spermatids (71.25%-93.75% ave=82.86%).

Every male with mean testicular length > 3.38 cm was producing long spermatids.

Possibly 10 animals with unilateral hypoplasia (8% with at least 0.5 cm [range between 0.5-0.7 cm] variation between R & L testes length).

Testicular length was chosen as the best measurement due to there being more variation with age. The width is not as variable between age groups. (1.59cm is difference between the mean testicular length age groups, 0.44cm is the difference between the mean testicular width age groups) There was also a good correlation between testicular length and the presence of long spermatids.

Discussion:

The intention of this research was to discover a good practical method for predicting the fertility potential in male alpacas. While there was a correlation between testicular width and the percentage of long spermatids visualised, the correlation between testicular length and percentage long spermatids was stronger. The correlation was still not as strong as expected After reviewing the data, it was found that alpacas are much more varied than other production animals when it come to body size and testicular size relating to each other or age or to the types of cells present in the testicles. This study has found the best correlation to be between testicular length and percentage of long spermatids (r2=0.49). Some general cut off measurements can be used to make management decisions. This is useful, as just with rams and bulls, alpaca testicles can be measured as part of a breeding soundness exam.

Some general cut off measurements can be used to make management decisions also. It was demonstrated that all animals with testicular length exceeding 3.38cm had long spermatids.  Further in this cohort, all animals over 3.2 years of age were producing long spermatids, supporting historical choices to mate 3 year old males (Sumar, 1996)

Ideally, young alpacas with longer testicles should be selected for breeding. This is a practical in vivo measurement which can be used by veterinarians and experienced alpaca farmers alike.

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