Use of Silver Nitrate for the Assessment of Sperm Measurements in Selected Farm and Free-Living Animal Species

Abstract

The study was conducted on spermatozoa of selected farm and free-living animal species, isolated post mortem from the tail of the epididymis, and stained with silver nitrate -AgNO3. The material was collected from pigs, goats, wild boar, and European roe deer. Twenty morphologically normal spermatozoa randomly selected from each animal and well visible under the microscope, were analysed. The following measurements were considered: head length, width, perimeter and area, acrosome area, mid-piece length, tail length, and overall sperm length. AgNO3 staining differentiated the acrosomal (light hue) and distal (dark hue) part of the sperm head, and a light-hued mid-piece was visible within the sperm tail. Silver nitrate staining revealed species and variety-related differences, particularly in reference to the sperm head. Clear-cut differentiation within the head and tail area made it possible to perform detailed morphometric measurements of the spermatozoa.

Differences in drone sperm morphometry and activity at the beginning and end of the season

“…Staining with silver nitrate also shows the heterogeneous structure of the sperm tail, which is probably linked to variation in the activity of mitochondria or mitochondrial derivatives. The use of silver nitrate staining to identify the sperm midpiece in other species has been described by Andraszek et al (13,14). A study by Collins and Donoghue (6) confirmed that it is useful to stain drone sperm cells using various techniques, because each technique can reveal different details of the sperm ultrastructure or defects in its morphological structure.…”

Comparison of the structure of chinchilla sperm isolated from semen and from the tail of the epididymis

“…As silver nitrate is an alkaline dye, it is mainly used to identify acidic chromatin proteins and the chromatin of nucleolus organizer regions in mitotic chromosomes (Andraszek, Horoszewicz, & Smalec, ), as well as nucleoli during meiosis (Andraszek, Gryzińska, Wójcik, Knaga, & Smalec, ; Andraszek, Szeleszczuk, Niedbała, & Kuchta‐Gładysz, ). The basic methodology has been modified in our Department (Department of Animal Genetics and Horse Breeding, University of Natural Sciences and Humanities in Siedlce) and successfully used to identify morphological details of spermatozoa from fresh, frozen or fixed semen samples, from mammals (Andraszek, Banaszewska, et al, ; Andraszek & Smalec, ; Banaszewska, Andraszek, Czubaszek, et al, ; Banaszewska, Andraszek, Szostek, et al, ; Banaszewska, Andraszek, Zdrowowicz, et al, ) birds (Andraszek et al, ) and insects (Gontarz, Banaszewska, Gryzińska, & Andraszek, ).…”

“…The use of silver nitrate in the assessment of sperm morphology and morphometry enables identification of structures that cannot be seen when other staining techniques are used (Andraszek Banaszewska & Biesiada‐Drzazga, ; Andraszek, Banaszewska, et al, ; Andraszek & Smalec, ; Banaszewska, Andraszek, Czubaszek, et al, ; Banaszewska, Andraszek, Szostek, et al, ; Banaszewska, Andraszek, Zdrowowicz, et al, ). Sperm stained with AgNO 3 are coloured in different degrees in the head and tail, with a clearly visible midpiece.…”

“…Previous research on semen has shown that sperm morphology is quite heterogeneous within a single ejaculate. Sperm are found to differ in size and slightly in shape, while having normal morphological structure (Andraszek, Banaszewska, et al, ; Andraszek, Gryzińska, Danielewicz, Batkowska, & Smalec, ; Banaszewska et al, ; Banaszewska, Andraszek, Szostek, et al, ; Banaszewska et al, ; Banaszewska et al, ; Banaszewska Kondracki & Wysokińska, ; Gago et al, ). A study by Cummins and Woodall () showed the following dimensions for chinchilla sperm: head length approximately 3 μm, midpiece length 4 μm, tail length 38 μm and total sperm length 46 μm.…”

“…Mammalian sperm are distinguished by their small size and compact structure, which sometimes makes it difficult to see the details of their morphology with a light microscope (Andraszek & Smalec, ). A search for techniques enabling the most accurate assessment of semen in terms of its functional features has revealed that a simple method of staining slides with silver nitrate captures more details of sperm structure than other routine methods (Andraszek, Banaszewska, & Biesiada‐Drzazga, ; Andraszek, Banaszewska, et al, ; Andraszek & Smalec, ; Banaszewska, Andraszek, Czubaszek, et al, ; Banaszewska, Andraszek, Szostek, et al, ; Banaszewska, Andraszek, Zdrowowicz, et al, ; Czubaszek, Andraszek, Banaszewska, & Walczak‐Jędrzejowska, ). Therefore, the aim of the work was comparison of the structure of chinchilla sperm isolated from semen and from the tail of the epididymis after staining with silver nitrate.…”