Int. J. Morphol., 31(3):1076-1080, 2013.

 

Morphological, Stereological and Histometrical Assessment of the Testicular Parameters between Holstein and Simmental Bulls

 

Evaluación Morfológica, Estereológica e Histométrica de los Parámetros Testiculares entre Toros Holstein y Simmental

 

Murat Sirri Akosman*; Ömer Faruk Lenger** & Hasan Hüseyin Demirel***

* Department of Anatomy, Faculty of Veterinary Medicine, Afyon Kocatepe University, Afyonkarahisar, Turkey.

** Department of Medical Biology and Genetics, Faculty of Veterinary Medicine, Afyon Kocatepe University, Afyonkarahisar, Turkey.

*** Department of Pathology, Faculty of Veterinary Medicine, Afyon Kocatepe University, Afyonkarahisar, Turkey.

Correspondence to:

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SUMMARY: The testicular measurement is important criteria for experimental researches especially in toxicological studies and the prediction of spermatogenesis. In light of this knowledge, we aimed to estimate and compare these parameters in two different kinds of cattle breeds. The gross anatomical measurements were performed by vernier caliper, volume of the testis was estimated by Cavalieri method and seminiferous tubule diameter was measured on the histological sections by software-loaded computer attached to microscope. The mean testis weight, length, width, volume, and tubule diameter of the Simmental bulls and the Holstein bulls measured as 316 g, 12.1 cm, 6.9 cm, 295 cm3 and 226.68 µm and 299 g, 12.1 cm, 6.8 cm, 285 cm3 and 223.44 µm, respectively. In conclusion all investigated parameters were not found statistically important in groups and between the breeds (p>0.05). The authors believed that the obtained data will contribute to the literature and facilitate future research.

KEY WORDS: Bull; Cavalieri method; Seminiferous tubule; Testis; Volume.

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RESUMEN: La medición testicular es un criterio importante para las investigaciones experimentales sobre todo en los estudios toxicológicos y predicción de la espermatogénesis. El objetivo fue estimar y comparar estos parámetros en dos diferentes tipos de razas de ganado. Las mediciones anatómicas fueron realizadas con un pie de metro, el volumen de los testículos se estimó por el método de Cavalieri y el diámetro de los túbulos seminíferos se midió en las secciones histológicas observadas al microscopio mediante software. La Media del peso testicular, longitud, ancho, volumen y diámetro de los túbulos seminíferos de los toros Simmental fueron 316 g, 12,1 cm, 6,9 cm, 295 cm3 y 226,68 µm y de los toros Holstein fueron 299 g, 12,1 cm, 6,8 cm, 285 cm3 y 223,44 µm, respectivamente. En conclusión, todos los parámetros investigados no tuvieron una importante significación en los grupos y entre las razas (p> 0,05). Creemos que los datos obtenidos contribuirán a la literatura y facilitar las futuras investigaciones.

PALABRA CLAVE: Toro; Método de Cavalieri; Túbulos seminíferos; Testículo; Volumen.

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INTRODUCTION

In Turkey, the Simmental and Holstein bulls are raised for the brood stock and beef aimed. The Turkish breeders keep Simmental cows for their beef and dairy features and the Holsteins cows for their highest-milk production. The adaptation of the Simmental cattles to the environment better than the Holstein cattles (Alpan & Arpacik, 1998). Actually, testicular volume measurement is a significant criterion in spermatogenesis prediction, evaluating testis functions and detected variety of disorders affects testicular growth and development (Sakamoto et al., 2007; Karaman et al., 2005; al Salim et al., 1995; Paltiel et al., 2002; Setchell & Brooks, 1988; Wing & Christensen, 1982; Sakr & Shalaby, 2012; Al-Damegh, 2012). Because 70-80% of the testis consist of seminiferous tubules and any change in testis measures reflects the changes in this tissue mass and also spermatogenesis (al Salim et al.; Paltiel et al.; Setchell & Brooks). As a result of these scientific findings, testicular and scrotal measurements are especially in common use as an important factor for the bulls to decided whether brood stock in the herd (Hahn et al., 1969; Silva et al., 2011).

Recently, there are some measurement techniques, including caliper (Gouletsou et al., 2008), orchidometry (Paltiel et al.), ultrasonography (Gouletsou et al.; Paltiel et al.) and the combination of MR-I and stereological volume estimation tool "Cavalieri method" (Coronado et al., 2011; Kabay et al., 2009; Gocmen-Mas et al., 2010) for in vivo testis volume measurements, and furthermore water displacement method "Archimedean principle" and individual administration of Cavalieri method (Akosman & Özdemir, 2010; Ince et al., 2012; Dorostghoal et al., 2011) for in vitro testis volume measurements. The volume estimation method "Cavalieri method" (Gundersen et al., 1988, 1999) is unbiased and current way to estimate the volume and volume fractions of the organ (Gundersen et al., 1988). If we mentioned the method simply, first of all the testis has to be sliced constant and in parallel form with a random start and then surface area of the obtained slices measured by a transparent test probe consist of the constant distance of points (Evans et al., 2004). The morphological and histological parameters mentioned above are really important parameters especially on the determination of the future reproduction ability of the herd. In this study, we aimed to investigate and compare these parameters between two cattle breeds of almost same age and same weights. We believe that the data will contribute and be useful for the future scientific researches.

Material and Method

This study was performed on 6 Simmental bulls (1-1.5 years old, 400-350 kg) and 6 Holstein bulls (1-1.5 years old, 300-350 kg) paired testis. All testes were removed immediately from their carcass after slaughter in the abattoir.

Tissue Preparation and Sampling Protocol. Firstly, the epididymis were removed and leaving only the testis proper and all testes were weighted (Kern, Balingen-Germany). The testis length (from proximal to distal pole) and the width were measured by vernier caliper (Labomar, 304B-01- Turkey). The important point on the width measurements, the testicular shape will affected from the resting on a table and avoid to distortion of testicle shape all testicles leaved to the water and gently hold by two pens from both poles and measured. After these measurements, neutral buffered 10% formalin immersion fixation performed for one week to testis.

Volume Estimation. The testes were sliced into 1 cm intervals from a random start and the areas per fractions on the slices were measured by 1 cm point grid.

The total testis volume was calculated by the formula below:

V= t x a/p x P cm³
t; mean slice thickness (1cm),
a/p; area per point (1cm x 1cm),
; counted points.

The coefficient of error (CE) of the volume estimations was calculated according to Nyengaard (1999).

Seminiferous Tubule Diameter Measurements. The little tissue pieces were obtained from the different regions of the testes, embedded in paraffin, sliced in 6 µm and stained by Haematoxylin and Eosin. The diameter of the seminiferous tubules were measured by Imagefocus software which was running on a computer connected to a 3.2 mp Cmex camera (Euromex) attached to Olympus BH2 microscope (Orhan et al., 2004; Tas et al., 2010; Yenilmez et al., 1995).

Statistics. All evaluated parameters in groups and between the breeds were compared using Mann-Whitney U statistical test.

Results

According to the morphological inspection the Simmental bulls were larger than the Holstein bulls even on the same age because of the Simmental cattle beef feature and their testes were in healthy condition. The mean testis weight, length and width were 316 g, 12.1 cm, 6.9 cm and 299 g, 12.1 cm, 6.8 cm for the Simmental and Holstein bulls, respectively (Table I).

Table I. The weight, length and the width of the testes.

According to stereological volume estimation the average testis volume of the Simmental bull was 295 cm3 while this value for the Holstein bull was 285 cm3 (Table II).

Finally, the mean diameter of the testicular tubules measured as 226.68 µm and 223.44 µm for the Simmental and Holstein bulls, respectively (Fig. 1, Table II).

There were no statistical significance found in groups and between the sexes (p>0.05) of the measurements of the testicular weights, lengths, widths, volumes and tubule diameters.

Coefficient error of the study. The CE value of the Cavalieri method was calculated as 0.02 for the Simmental bull testis and 0.02 for the Holstein bull testis.

Table II. The average volume of the testis, coefficient of error of the
Cavalieri method and the results of the seminiferous tubule diameter measurements.

Fig. 1. A histological view of the parenchyma of a A. Holstein bulls left testis,
B. Holstein bulls right testis, C. Simmental bulls left testis, D. Simmental bulls right testis.

Discussion

The in vivo testicular measurement could allow to estimate the spermatogenesis and testicular functions (Sakamoto et al.; Karaman et al.). The scrotal circumference, testicular volume and shape especially important for reproductive potential in young beef cattle as a selection criteria (Hahn et al.; Silva et al.). These dimensions highly correlated with the total sperm production (Hahn et al.). The measurements of these parameters could easily achieved using caliper in daily practice (Bailey et al., 1998). The scientists mentioned that the ultrasound imaging also use beneficially to estimate the size and volume of the testis (Bailey et al., 1998). The volume estimation technique here we applied, does not individually permit in vivo estimations in daily practice. However it is very successful and useful in the in vitro studies. It allows to estimate the total and components volume of the organ (Pazvant et al., 2009). Additionally, the Cavalieri methods coefficient of error value was found so appropriate. Generally less than %5 CE is an acceptable value for a study and shows the quality of the sampling procedure (Bolat et al., 2011; Pazvant et al.).

The result of all investigated parameters include seminiferous tubules, in groups and between breeds were not found statistically important (p>0.05). This may depend on animals of almost same age and same weights. However, in a study on 17-22 month-old Holstein bulls, the weights of the testicles and the lengths were measured as 460 g and 14.7 cm (Hahn et al.). This difference may originated from the animals age. Because testicular weight increase depends to the age (Hahn et al.). The measured length of the seminiferous tubule between the breeds has no statistical significance (Fig. 1). The measurement of the seminiferous tubule and volume estimation of the total testis volume will be valuable especially in the toxicological research and the determination of the environmental pollution. The environmental polluters such as pesticides and heavy metals causes disorders on the reproductive activity and the impairment on the germ cell in the testis of the mammals (Bonde, 2010; Fejes et al., 1991; Meldrum & Ko, 2003; Ince et al.; Michael et al., 2007; Bailey et al., 2004). In conclusion, in this research we estimated the testicular weight, length, width, volume and the diameters of the seminiferous tubules and could not detected statistically importance in groups or between sexes (p>0.05). This kind of measurements on the testis will be beneficial on the evaluation of the reproduction features of the herd, toxicological research and the testicular effects of the environmental polluters.

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Correspondence to:

Murat Sirri Akosman

Department of Anatomy
Faculty of Veterinary Medicine
Afyon Kocatepe University
Afyonkarahisar
TURKEY
Phone: +902722281312-232

Email: akosmans@yahoo.com

Received: 24-12-2012
Accepted: 19-05-2013