Understanding critical roles of warming and reanimation is critical to improve the survival of vitrified testicular tissue in domestic cats. The objective was to study structural and functional properties of testicular tissues from prepubertal domestic cats after standard vitrification followed by two warming protocols (directly at 37°C or with a 5-second pre-exposure to 50°C) and three reanimation time points (immediately, 24 h and 5 days post-warming). In Experiment 1, tissues were evaluated for histo-morphology and mitochondrial activity immediately or 24 h after warming protocols. In Experiment 2, cell viability, DNA fragmentation, and germ cell composition were assessed immediately, 24 h, or 5 days after optimal warming. Preservation of seminiferous tubule structure was better using warming at 50°C for five seconds, and survival of somatic as well as germinal cells was higher compared to direct warming at 37°C for one minute. Short term in vitro culture (for reanimation) also proved that cellular composition and functionality were better preserved when warmed for a short time at 50°C. Collective data showed that short warming at 50°C led to better quality of seminiferous tubule structure and cell composition after vitrification and short-term culture. In addition, data suggest clear directions to further understand and optimize testicular tissue survival after fertility preservation procedures.

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(2015) Semen quality, testicular B-mode and Doppler ultrasound, and serum testosterone concentrations in dogs with established infertility. Theriogenology, 84 (5). pp. 805-810. ISSN 1879-3231 Access from the University of Nottingham repository: http://eprints.nottingham.ac.uk/38920/1/Dogs%20with%20Established%20Infertility %20Theriogenology.pdf Copyright and reuse:The Nottingham ePrints service makes this work by researchers of the University of Nottingham available open access under the following conditions. This article is made available under the Creative Commons Attribution Non-commercial No Derivatives licence and may be reused according to the conditions of the licence. For more details see: http://creativecommons.org/licenses/by-nc-nd/2.5/ A note on versions:The version presented here may differ from the published version or from the version of record. If you wish to cite this item you are advised to consult the publisher's version. Please see the repository url above for details on accessing the published version and note that access may require a subscription. concentrations (1.40 ± 0.62 ng / mL) than fertile dogs (1.81 ± 0.87 ng / mL) (P < 0.05). 37There were subjective differences in testicular echogenicity in some of the infertile 38 dogs, and important differences in testicular artery blood flow with lower peak systolic 39 and end diastolic velocities measured in the distal supra-testicular artery, marginal 40 testicular artery and intra-testicular artery of infertile dogs (P < 0.05). Notably, 41resistance index and pulsatility index did not differ between infertile and fertile dogs. 42These findings demonstrate important differences between infertile and fertile dogs 43 which may be detected within an expanded breeding soundness examination.

(2015) Digital image analysis of testicular and prostatic ultrasonographic echogencity and heterogeneity in dogs and the relation to semen quality. Animal Reproduction Science, 160 . pp. 112-119. ISSN 1873-2232 Access from the University of Nottingham repository: http://eprints.nottingham.ac.uk/38918/1/Image%20analysis%20testes%20and%20prostate %20Animal%20Reproduction%20Science.pdf Copyright and reuse:The Nottingham ePrints service makes this work by researchers of the University of Nottingham available open access under the following conditions. This article is made available under the Creative Commons Attribution Non-commercial No Derivatives licence and may be reused according to the conditions of the licence. For more details see: http://creativecommons.org/licenses/by-nc-nd/2.5/ A note on versions:The version presented here may differ from the published version or from the version of record. If you wish to cite this item you are advised to consult the publisher's version. Please see the repository url above for details on accessing the published version and note that access may require a subscription.For more information, please contact eprints@nottingham.ac.uk shape, position, margination and internal architecture of the testes (England, 58 1991;Eilts et al., 1993; Paltiel et al., 2002;Gouletsou et al., 2008; Souza et al., 59 2014) and prostate gland (Blum et al., 1985; Juniewicz et al., 1989; England, 60 1991;Eilts et al., 1993; Ruel et al., 1998; Paltiel et al., 2002; Gouletsou et al., 61 2008; Freitas et al., 2013;Freitas et al., 2015). Ultrasonography also provides a 62 valuable tool in assessing reproductive pathology (Cartee and Rowles, 1983; 63 Feeney et al., 1987; Pugh and Konde, 1991; Cooney et al., 1992; England, 1995; 64 Keenan, 1998;Nautrup and Tobias, 2001; Hecht, 2008). 65 66In clinical practice, ultrasound images are subjectively assessed and described in 67 terms of their image texture; principally echogenicity and heterogeneity. A small 68 number of studies have proposed a relationship between grossly detectable 69 lesions within the testes and semen quality (England, 1991; Vencato et al., 2014). 70Objective analysis of echogenicity from measurements of pixel intensity is 71 however possible using digital image analysis (Ivancic and Mai, 2008) The aim of this study was to measure testicular and prostatic ultrasonographic 89 echogenicity and heterogeneity using digital image analysis, and investigate the 90 relationships between these measures and semen quality in a group of known 91 fertile dogs. 92 93 Materials and Methods 94 Study animals 95Forty-three stud dogs (21 Labrador Retrievers, 12 Golden Retrievers, 6 German 96 Shepherds, 1 Border Collie, 1 Flat Coated Retriever, 1 Irish Water Spaniel and 1 97Standard Poodle) with a mean weight of 35.5 ± 5.8 kg (range 20.6 to 54.1 kg) 98 aged between 1.1 and 9.3 years (mean 4.2 ± 2.0 years) were examined. Dogs 99 5 were selected on the basis that they met the following inclusion criteria; (1) 100 clinically healthy, (2) o...

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Companion Animals Ultrasonographic imaging is an important diagnostic tool because it allows assessment of the shape, size, position, margination and internal architecture of organs, as well as facilitating the study of vascular supply and vascularisation. Recently, there has been considerable development of B-mode, Doppler and contrast-enhanced ultrasonography for examination of the reproductive tract of dogs, both for studying normal physiology and in the clinical setting. This article describes the practical examination of the canine prostate gland and testes using a variety of ultrasound techniques, and details the normal appearance and blood flow of these organs as well as changes that may be observed with common reproductive disorders. Imaging the prostate gland An image of the prostate gland can be achieved by placing the ultrasound transducer on the caudoventral abdomen, adjacent to the prepuce. Most dogs can be examined without sedation while standing or when positioned in dorsal or lateral recumbency. A 7.5 to 10.0 MHz transducer is preferable and imaging is facilitated by the presence of urine within the bladder. Imaging should always be undertaken in both the transverse and longitudinal planes, while the frontal (dorsal) plane is also useful. When the prostate lies entirely within the pelvis, transrectal imaging can be performed in large dogs but not in small dogs. B-mode ultrasonography Normal prostate gland The prostate is an ovoid-shaped, bilobed gland positioned

Sperm morphometry is the tool that confers objectivity to the morphological evaluation by accurately measuring the dimensions of the gamete and its structures. Thus, the aim of the study was to perform a morphometric characterization of the domestic cat sperm. Therefore, sperm samples were collected from twenty pairs of epididymis in a TRIS extender at 37ºC. An aliquot of the sample was used to make a smear with Rose Bengal solution, and afterwards, the morphology and morphometry were analysed. In the morphology, were quantified the percentage of normal sperm cells, morphological changes of head, midpiece and tail. In morphometry, each normal sperm cell was measured for length, width, area and perimeter of head and midpiece, tail length and total length. The parameters ellipticity, elongation, regularity and rugosity were also determined. The percentage of normal sperm was 67.21%. Of the abnormalities, the curled/folded tail, followed by the curved midpiece, abnormal shaped head and detached head were the most quantified. The sperm head presented 5.56 ± 0.01 μm and 3.10 ± 0.01 μm of length and width, respectively. The head area was 16.94 ± 0.05 μm2, while the perimeter was 16.16 ± 0.03 μm. In the derived parameters, the values were as follows: ellipticity of 1.81 ± 0.00; elongation of 21.39 ± 0.12; regularity of 0.81 ± 0.00; and rugosity of 0.14 ± 0.00. The midpiece presented length and width of 7.96 ± 0.01 μm and 0.76 ± 0.01 μm, respectively. The mean length of the sperm tail was 45.12 ± 0.06 μm, and the total cell size was 58.67 ± 0.06 μm. Thus, it was concluded that the cat sperm is an elongated cell, with high rugosity and regularity. The spermatic tail represents more than ¾ of the total length of the cell and the midpiece exceeds the length of the head.