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Theriogenology 77 (2012) 459 465 www.theriojournal.com

Reference values for the genitalia of male dromedary before and after puberty using caliper and ultrasonography in subtropics
Derar Refaat Derar*, Hasan Ali Hussein, Ahmad Ali
Department of Theriogenology, Faculty of Veterinary Medicine, Assiut University, Assiut, 71526, Egypt Received 5 February 2011; received in revised form 6 August 2011; accepted 16 August 2011

Abstract The aim of the present study was to characterize the dynamic changes of the testes and the pelvic genitalia in vivo in male dromedary. Eighty-one clinically healthy male dromedaries aged 1.5 to 12 years were assigned for the present study. Testicular length, breadth, and depth as well as epididymal head and tail were measured using caliper and ultrasonography. The pelvic genitalia, including bulbourethral gland, prostate, and pelvic urethra were examined using ultrasonography. The results revealed that the three dimensions of the testes and epididymal tail and head showed signicant increase with age (P 0.01). Concerning the epididymal measurements, differences between the pre- and peri-pubertal groups were not signicant. Left testes tended to be larger than the right (not statistically signicant) although only the breadth of the left testes in the prepubertal group was signicantly larger (P 0.05). The volume of both testes correlated positively with the age (r2 0.91 for left and 1.00 for the right, P 0.01). There were no signicant correlations between the values measured using caliper and those by ultrasonography between groups, but the correlation was highly signicant (P 0.01) for the total number of the examined animals. There were signicant and steady increases of the size of bulbourethral gland in all examined groups (P 0.01). Pars disseminata of the prostate gland and pelvic urethra were signicantly higher in sexually mature compared with prepubertal groups (P 0.01). It was concluded that ultrasonography is a useful tool in studying the developmental changes of the testes and accessory glands of the male dromedary. The obtained data could provide a reference values for predicting camel puberty and future fertility. 2012 Elsevier Inc. All rights reserved.
Keywords: Male camel; Testes; Epididymis; Caliper; Ultrasonography

1. Introduction Although the camel is one of the oldest known mammals, studies on the different aspects of the physiology and pathology of reproduction in this species have been limited. However, in the last decades, interest in this species was renewed for fundamental and applied research in reproductive physiology [1]. Full physical examination of the male camel is essential

* Corresponding author. Tel.: 2-088-2334699; fax: 2232564. E-mail address: derar40@yahoo.com (D.R. Derar).

2-088-

before purchase to ensure its soundness. It is also advisable that male camels undergo regular clinical examinations at the beginning of each season to ensure that no problems have arisen that may reduce fertility. Age is an important aspect in considering the potential fertility of a camel. Young and old dromedary bulls may have problems with taking on a full breeding schedule with consistent success rates. It would be valuable to predict the age at which a camel would reach puberty and its future fertility based on the developmental parameters measured in the camel genitalia. Ultrasonography has been used to study reproductive tract and testes development in bull calf [2,3] and

0093-691X/$ see front matter 2012 Elsevier Inc. All rights reserved. doi:10.1016/j.theriogenology.2011.08.023

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D.R. Derar et al. / Theriogenology 77 (2012) 459 465 0.18a 0.23b 0.11c 1.55 2.59 3.15 1.53 (n 27) 46 (n 25) 712 (n 29) 3.69 6.59 7.83 0.31a 0.19b 0.5c 2.09 2.91 3.65 0.16a 0.11b 0.07c 2.47 2.75 3.62 0.19a 0.05a 0.25b 0.24a 0.15b 0.08c Means with different superscripts in the same column are signicantly different (P TL, TB, TD, testicular length, breadth, and depth. 0.01). 3.69 6.26 7.65 0.26a 0.23b 0.52c 1.87 2.76 3.45 0.23a 0.07b 0.07c 2.24 2.63 3.51 0.14a 0.04b 0.15c 1.49 2.41 3.00

SEM).

Right testes Left testes Age (year)

2. Materials and methods 2.1. Animals Eighty-one male dromedary camels aged 1.5 to 3 years (n 27, prepubertal), 4 to 6 years (n 25, peri-pubertal) and 7 to 12 years (n 29, sexually mature) were used for the caliper and ultrasonographic examinations. Animals belonged to 12 small stockholders in El-Minia and Assiut provinces, Egypt (latitude: 8.2 meters, 25.4 centimetres, 58 N; longitude 10.1 meters, 25.4 centimetres, 58 E; altitude 37 m. All animals were clinically healthy, free from external and internal parasites, fed ad libitum on maintenance rations according to requirements described by NRC for each age category, and lived under the same management and prevalent environmental conditions. 2.2. Morphometric and ultrasonographic examination All examinations were performed in sternal recumbency and the camels were secured with ropes from the fore to hind limbs. Testicular length (TL), Breadth (TB), and depth (TD) were measured by caliper at their maximum dimensions nearly at the same point for each animal. At the same time, the scrotal contents, including testes and epididymis, were examined using a 6/8 MHz linear array transducer connected to B-mode ultrasound scanner (100 LC, Pie Medical, Masstricht, the Netherlands). The testes were scanned in transverse and longitudinal planes. The thickness of tunica albugenia and mediastinum testis were evaluated from a longitudinal plane. The head and tail of the epididymis were measured using both the caliper and ultrasonic device and differences between groups were recorded. The head of the epididymis was visualized ultrasonographically from a diagonal plane near the ventral end of the testis (oblique axis testes and the epididymis faces laterally and situated along the dorsal border of the testes). The pelvic genitalia, including bulbourethral gland (BUG), pelvic urethera, pars disseminate, and body of the prostate were also scanned. Images were frozen on the monitor of the ultrasound scanner and

Table 1 Variation in testicular length, depth (using caliper) and breadth (using caliper and ultrasonography) relative to the age of the male dromedary (mean

TB (cm) TL (cm) TD (cm) TB (cm) TL (cm) Caliper Caliper Sonar Caliper Caliper Caliper

Sonar

in male camels [4]. The objectives of this study were to use caliper and ultrasound to assess the developmental changes in the testes, epididymis, prostate, and bulbourethral gland (BUG) in prepubertal, peri-pubertal, and sexually mature male camels. Moreover, the different parts of the male dromedary external and internal genitalia were measured to establish reference values for future pre-purchase and soundness examinations.

TD (cm)

Caliper

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Fig. 1. Ultrasonographic image of the whole testes of a young camel (1.5 years).

Fig. 3. longitudinal image of camel testis (6 years) MT, mediastinum testis; Tail., tunica albugenia.

dimensions of these organs were measured at their maximum. 2.3. Statistical analysis After recording the actual testicular length, breadth, and depth, the volume was calculated according to the formula for the prediction of testicular volume [5,6]. The predicted volume of each testis was calculated by the formula for a prolate spheroid [5,7,8]: V 4/3( )(L/2)(W/2) [2], where L equals the length of the testis, and W is the widest diameter of the testis. This formula reduced to: V 0.5236(LW2). The product of the calculations from the formula for a prolate spheroid was then tested against the actual volume for verication of the predicted volume of each testis by the least squares method of tting a regression line to the data [6]. For the regression lines, the equation is in the

standard line form: y a bx, where b is the slope and a is the y-intercept. To correlate testis measurements for volume, an independent regression analysis for the calculated volume for each testis collected [6] and linear regression was drawn for the calculated volume and age. The difference between caliper and ultrasonographic measurements, right and left testes were determined by comparing means using paired-samples t-test. The correlation between the obtained measurements and the age was determined using correlation bivariate [6]. 3. Results 3.1. Testes The TL, TB, and TD estimated by caliper for the 81 camels are presented in Table 1. Age had a signicant

Fig. 2. ultrasonographic image of part of camel testes (4 MT, mediastinum testis; Tail., tunica albugenia.

5 years)

Fig. 4. The tail of the epididymis image, oblique plane (6 years), Te., Testis; ET, epididymal tail.

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Table 2 Variation in tunica albugenia, mediastinum of the left and right testes in relation to the age of the male dromedary as estimated ultrasonographically (mean SEM). Age (year) Left 1.53 (n 27) 46 (n 25) 712 (n 29)
a,b

Tunica albugenia (cm) Right 0.23 0.25 0.29 0.01a 0.01a 0.01b 0.01). 0.22 0.28 0.34 Left 0.20 0.22 0.30 0.0a 0.02a 0.01b

Mediastinum testis (cm) Right 0.22 0.26 0.39 0.03a 0.02a 0.02b 0.03a 0.01b 0.01b

Means with different superscripts in the same columns are signicantly different (P

effect on all testicular measurements estimated by caliper (P 0.01). The TL was estimated easily at all ages using the caliper, but it was difcult using ultrasonography as the increasingly developed testicular length was beyond the scope of the transducer. Left testis was non-signicantly (except for the TB in the premature group, P 0.05) larger than the right one. Testicular parenchyma showed increased echogenicity with age (Figs. 13) and a transverse image of the testis (Fig. 3) showed the mediastinum as hyperechogenic spot in the middle. The volume of both testes correlated positively with age for left (r2 0.9059, P 0.01) and right (r2 1.000, P 0.01) testes. There were no signicant correlations between the measurements taken by caliper and ultrasonography between groups, but the correlation was highly signicant (P 0.01) for the total number of the examined animals. A transverse image of the testis (Fig. 4) showed the mediastinum as hyperechogenic spot in the middle. The thickness of the tunica albugenia and mediastinum are presented in Table 2. Tunica albugenia showed slow rate of growth in all the examined groups while the mediastinal growth rate was slower in the prepubertal and peri-pubertal groups than in the sexually mature group. Thickness of the tunica albugenia and mediastinum testis estimated by ultrasonography was signicantly affected (P 0.01) by age.

3.2. Epididymis The head of the epididymis image was recorded from the oblique plane near the distal pole of the testis. It is well differentiated from the testicular parenchyma as a triangular-like area separated by a non-echogenic line and has a less echogenic appearance (Fig. 4). Age affected the epididymal measurements signicantly (Table 3, P 0.01). The differences between groups were statistically signicant in all measurements of the left and right epididymal heads except in the premature group (ultrasonography, P 0.001). There were no signicant differences between the left and right epididymal tail measurements (caliper) in all examined groups. There were signicant correlations between the measurements taken by caliper and those of ultrasonography in the peri-pubertal group and the correlation was signicant (P 0.05) for the total number of the examined animals. 3.3. Pelvic genitalia The pars externa were paired hyperechogenic structures (Fig. 5) while BUG appeared as fusiform uniformly hyperechogenic structure embedded in the bulbocavernous muscle (Fig. 6). Age had a steady and signicant effect on BUG, pars disseminate, length (not the breadth) of the pars externa of the prostate, diameter of the pelvic urethra measurements as estimated ultra-

Table 3 Variation in epididymal head (caliper and sonar) and tail (caliper) diameter in relation to the age of the male dromedary (mean Age (year) Left (cm) Caliper 1.53 (n 27) 46 (n 25) 712 (n 29)
a,b

SEM).

Epididymal Head Right (cm) Sonar 1.34 1.62 1.72 0.11a 0.07b 0.04c Caliper 1.53 1.69 2.03 0.08a 0.13a 0.08b Sonar 1.22 1.36 1.62 0.14a 0.07b 0.04b

Epididymal tail Left (cm) Caliper 1.48 1.78 2.45 0.16a 0.13a 0.09b Right (cm) Caliper 1.36 1.67 2.33 0.17a 0.13a 0.11b

1.69 1.89 2.18

0.06a 0.16a 0.05b

Means with different superscripts in the same columns are signicantly different (P

0.01).

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Fig. 5. Prostate of the adult camel, (A) pars externa, (B) pars disseminata, (C) diameter of pars disseminata PU., pelvic urethra (12 years old camel).

sonography (Table 4, P 0.01). However, ultrasonographic values of the pars disseminate did differ between peri-pubertal and sexually mature groups. The disseminated part of the prostate appeared as a hyperechogenic area between the non-echogenic urethral lumen and less echogenic urethral muscle. 4. Discussion The study of the developmental changes of the male camel genitalia using ultrasonography is important to predict their future reproductive capacity and establish reference values for the developmental changes in their dimensions. The results of the present study indicated that a signicant increase in the testicular dimensions (TL, TB, and TD) in adult male camels compared to younger ones was found. These results agree with those recorded previously [9,10,11]. In the present work, it was clear that the left testicular measurements were greater than the right similar to previous results [4]. The preference of left side in male and female camel repro-

ductive functional anatomy and physiology is questionable and yet unexplainable. In the female, despite the even distribution of cyclic activity between the right and left ovaries, most of pregnancies, if not all, take place in the left horn. Moreover, left uterine horn is larger than the right even in the camel embryo [21]. Factors like blood vasculature and innervation should be investigated in this regard [23]. Testicular dimensions measured by caliper and testicular breadth estimated from ultrasonographic image are quiet reliable means for its growth follow-up. The whole testis was imaged using ultrasonography in the young animals. The caliper is easier to use and demonstrate a higher degree of accuracy for measurement of testicular length. Further work is needed to alleviate the problems encountered in determining testicular length by ultrasonography [8]. Testicular parenchyma showed changes in the echogenicity of its ultrasonic image during development. This is attributed to cellular proliferation and uid production. At the same time, an increase in testicular echodensity as male camels

Fig. 6. Bulbourethral gland (BUG), PU., Pelvic urethra (7 years old camel).

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Table 4 Changes in the maximum diameters of accessory genital glands in relation to the age as estimated from ultrasonographic image (mean SEM). Age (year) Left (cm) 1.53 (n 27) 46 (n 25) 712 (n 29)
a,b

BUG Right (cm) 1.13 1.38 1.74 0.05a 0.06b 0.8c Prostate gland Pars diss (cm). 0.80 1.22 1.48 0.03a 1.22b 0.13b

Pars externa Length (cm) 2.61 2.94 3.32 0.09a 0.9b 0.33b 0.01). Breadth (cm) 1.73 1.78 2.05 0.14a 0.15a 0.27a

Pelvic diameter (cm)

1.06 1.33 1.67

0.01a 0.11b 0.10c

2.13 2.37 2.62

0.10a 0.09b 0.22b

Means with different superscripts in the same column are signicantly different (P

achieved puberty agrees with previous results [2,22]. After puberty, the mediastinum width showed signicant increase concomitant with the increase in the testicular breadth supporting previous studies [1214] and contradictory to others [15] for cattle bull. Differences in breed studied, season, nutrition, body condition scoring and other management factors may be crucial factors on this respect. The formula for a prolate spheroid and its modication provides a rapid and highly reliable method for estimating testicular volume from testicular length and width measurements. The advantages of length and width measurements and application to a simple mathematical formula are readily apparent. This formula utilizes real physical measurements of individual testicular length and width, unlike that of Scrotal Circumference (SC) which measures only one dimension and encompasses both testicles simultaneously. The formula would be useful in following an animal through a long-term course of study for an accurate assessment of single testicle volume changes with time. Bulls are often presented with a variation in testicular shape [16 18). Frequently seen are the long cylinder testicles, rounded testicles, and normal ovoid scrotum [16 18]. The physical measurement of length and width can now be considered in the calculation of total testicular volume, rather than a single measurement of SC, or testicular width, which may not reect the total volume. Since individual testis can be measured and the formula applied, variation in testicular shape among individual bulls is discounted as a potential problem in calculating total volume of the two testes. The tail of the epididymis was easily estimated from oblique plane as a characteristic triangular hyperechogenic structure, which is separated from testicular parenchyma with non-echogenic line. The head was not easy to diagnose as in previous studies [19]. Using transrectal ultrasonography, Prostate gland, pars dissemenata, pelvic urethra and bulbourethral glands were visualized and their different measurements were recorded. This possibility cannot be carried through rectal palpation not

only because of soft consistency of accessory genital glands but also because of the inaccessible location of the BUG and disseminate prostate [20]. In the present study scrotal length and circumference were not measured in all examined groups because it is not pendulous and tightly adhered to the thigh under the anus. The current study indicated that ultrasonography is applicable in diagnostics, and gives appreciable benets in studying the developmental changes of the testes and accessory glands. But, it added no advantage over that of calipers for measuring the length of the testes in vivo except when only testicular breadth is needed. The growth of the accessory glands depends on testicular growth. Moreover, reference values for estimating developmental changes in the male dromedary in subtropics considered a useful tool for predicting male camel fertility. References
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