Measurement of Alkaline Phosphatase in Canine Seminal Plasma – An Update

In dogs, diagnosis of incomplete ejaculation and azoospermia can be made by measuring the activity of the enzyme alkaline phosphatase (AP) in seminal plasma. However, even though upper cut‐off value of 5000 IU / l is given in the literature, results by different assays may vary considerably. Furthermore, no data exist concerning the stability of the enzyme during storage of frozen seminal plasma, and no recommendations for pre‐analytic dilutions can be found. During the present study, we compared results from a conventional large scale wet chemistry analyzer to a widely used dry chemistry point of care system (POC) and established a best practice for pre‐analytical dilutions. Furthermore, stability of enzyme activities in seminal plasma during storage at ?18°C for 24 h was evaluated. The average activity of AP in the 2nd fraction of normal ejaculates measured by Reflotron® was 107 328 IU / l. After 24 h of frozen storage, activities did not differ significantly (96 844 IU / l, p > 0.05). Fresh and frozen samples were analysed in parallel by the POC and conventional chemistry analyser, and the results compared that did not reveal a significant difference (p > 0.05). A dilution of seminal plasma with physiologic saline 1 : 100 prior to analysis was sufficient for the qualitative information whether AP activity is below or above 5000 IU / l. Present data show that AP measurement by a POC dry chemistry system is sufficiently accurate in diluted seminal plasma for the diagnosis of azoospermia and that seminal plasma can be stored frozen for 24 h before analysis.     

Pharmacokinetics of an extended‐release formulation of eprinomectin in healthy adult alpacas and its use in alpacas confirmed with mange

The study objective was to determine the pharmacokinetics and clinical effects of an extended‐release 5% eprinomectin formulation (Longrange^®) following subcutaneous (s.c.) injection in healthy (n = 6) and mange‐infected (n = 4) adult alpacas. High‐performance liquid chromatography was used to analyze plasma samples obtained at regular intervals for 161 days following a single 5 mg/kg injection s.c. in healthy alpacas, and for 5 days following each dose (3 treatments, 2 months apart) in mange‐affected animals. Skin scrapings and biopsies were performed pre‐ and post‐treatment at two comparable sites in alpacas with mange. Four alpacas served as healthy controls. Eprinomectin plasma concentrations showed a biphasic peak (C_MAX‐1: 5.72 ± 3.25 ng/mL; C_MAX‐2: 6.06 ± 2.47 ng/mL) in all animals at 3.88 ± 5.16 days and 77 ± 12.52 days, respectively. Eprinomectin plasma concentrations remained above 1.27 ± 0.96 ng/mL for up to 120 days. Hematocrit (35.8 vs. 31.3%, P < 0.003) and albumin (3.5 vs. 2.8 g/dL P < 0.006) reduced significantly over 6 months in multidose animals, while fecal egg counts did not differ between groups. Self‐limiting injection site reactions occurred in 9 of 10 animals. Pre‐ and post‐treatment skin biopsies showed reduced hyperkeratosis, but increased fibrosis, with 1 of 4 alpacas remaining positive on skin scraping for mange. In conclusion, alpacas require a higher eprinomectin dose (5.0 mg/kg s.c.) than cattle, to reach comparable plasma concentrations.     

Seasonal variation in testicular blood flow dynamics and their relation to systemic and testicular oxidant/antioxidant biomarkers and androgens in rams

The environmental temperature increased during summer and decreased during winter to the limits that might negatively affect animal and human reproduction. The responses of Egyptian rams to either hot or cold climatic conditions were studied in six mature rams subjected to weekly testicular Doppler ultrasonographic examination, blood sampling, seminal plasma collection and semen evaluation. The maximum environmental temperature and the relative humidity were used to classify the climatic condition according to the heat stress equation of sheep into hot months where temperature–humidity index (THI) was >26 (31.67 ± 0.54), and cold months where THI was <22 (18.39 ± 0.41). Testosterone, estradiol, superoxide dismutase (SOD), glutathione peroxidase (GPX) and lipid peroxide product (malondialdehyde, MDA) were measured in both blood and seminal plasma, while catalase (CAT) and reduced glutathione (GSH) were measured in blood and seminal plasma, respectively. Results revealed that, during the hot months, rams displayed significantly decreased testicular blood flow, increased seminal plasma MDA, decreased seminal plasma (SOD, GPx and GSH) and blood CAT antioxidant enzymes. The present study evidenced two novel findings: (a) the marked decrease in testicular blood flow volume, that is remarkable increase in both resistive index (RI) and pulsatility index (PI) values, during hot months could be negatively affected both seminal plasma enzymatic activities and seminal attributes, and (b) the SOD and GPx activities in seminal plasma of such animals were suitable predictive markers for seminal attribute evaluation.     

The Origin of Membrane Vesicles in Ram Seminal Plasma

The hypothesis tested in this study was that the membrane vesicles present in ram seminal plasma are of testicular origin, rather than being secreted by the accessory sex glands as has been previously reported for a number of species. Membrane vesicles were present in cellular extracts from reproductive organs and accessory sex glands of six rams, and in the seminal plasma of a further eight rams. When four of the latter rams were subjected to vasectomy, to isolate ejaculate contents to only the secretions of the accessory sex glands, the vesicles were largely eliminated from their ejaculates, while vesicles were still present in the ejaculates of the four control rams. The constituents of the cytoplasmic droplets and membrane vesicles derived from the seminal plasma were compared by transmission electron microscopy (TEM). Vesicles present in the cytoplasmic droplets were similar in morphology but smaller on average than those in the seminal plasma. It was concluded that the membrane vesicles in ram seminal plasma originate from either the cytoplasmic droplets, or a combination of vesicles from the droplets and the epididymis.     

Effect of testicular tissue lysate on developmental competence of porcine oocytes matured and fertilized in vitro

The objective of the present study was to investigate the effect of testicular tissue lysate (TTL) on developmental competence of germinal vesicle (GV) stage porcine oocytes. Two types of TTL were prepared through repeated freeze–thaw in liquid nitrogen, one from whole testicular tissue (wTTL) and other from either of four different sections of testes, namely just beneath the tunica albuginea (TA), from the transitional area between the seminiferous cord/tubules and the mediastinum testis (TR) and from the intermediate area (parenchymal tissue origin) and CE (cauda epididymis origin). The whole or section‐wise TTL treatments were given for 44 hr during in vitro maturation (IVM). Oocyte maturation was done in either of the two media, namely defined (high‐performance basic medium for porcine oocyte maturation, commercially available) and serum containing (TCM199). After maturation, oocytes were co‐incubated with fresh spermatozoa for 6 hr and then transferred to embryo culture media. Treatment of GV stage oocytes with wTTL (1 mg/ml) increased the cleavage and morula percentage rate (69.23 ± 6.23 and 48.15 ± 6.77, respectively) than that of their control (58.33 ± 8.08 and 32.54 ± 5.53, respectively) in defined media, and in serum‐containing media, cleavage and morula percentage rate were almost equal in both treatment (54.56 ± 7.79 and 34.70 ± 6.78, respectively) and control (59.52 ± 8.21 and 38.52 ± 6.54, respectively). However, effect of wTTL was not significant. In case of section‐wise TTL supplements, TR section significantly (p < .01) improved cleavage and morula rate (58.43 ± 7.98 and 36.14 ± 6.89, respectively) followed by TA. In conclusion, present study indicates that IVM, in vitro fertilization and in vitro culture of embryo are improved in the presence of TTL, particularly its TR section. Further study is expected to reveal the principal components of TTL which may prove useful for IVM.     

The relationship of plasma leptin concentration and puberty in Holstein bull calves (Bos taurus)

The objective of this experiment was to study the changes of plasma leptin concentration during puberty and its relationship with testosterone level and testis dimensions in Holstein bull calves. Six Iranian Holstein bull calves with approximately 6 months of age were used. Semen evaluation was conducted at 1‐month interval to determine the puberty state. To detect the plasma leptin and testosterone changes, blood samples were collected from the jugular vein during pre‐puberty (6–7 months of age), puberty (8–9 months of age) and post‐puberty (10–11 months of age). In addition, body weight (BW), body condition score (BCS) and testicular width and length were measured at 3‐week intervals. The effects of time (age) on total sperm number and percentage of progressive motility of sperm, plasma concentration of leptin and testosterone, amplitude and frequencies of testosterone, BW, BCS, testicular dimensions were significant. Sperm number and progressive motility during post‐puberty were higher than those during puberty and pre‐puberty. Plasma concentration of leptin during the pre‐puberty was higher than those during puberty and post‐puberty (p < 0.01). Mean plasma testosterone concentrations during puberty were higher than those during pre‐puberty (p < 0.05). BW, BCS and testicular dimensions consistently increased throughout the trial. Results indicated that in growing bull calves, plasma concentrations of leptin decreased during puberty, while circulating testosterone increased.     

Reproductive Development of Santa Inês Rams During the First Year of Life: Body and Testis Growth,Testosterone Concentrations,Sperm Parameters,Age at Puberty and Seminal Plasma Proteins

We have investigated the reproductive development of the tropically adapted Santa Inês ram, the most common hair sheep in Brazil. From 8 to 48 weeks of age, 16 animals were evaluated for body and testis growth, semen parameters, testosterone concentrations and seminal plasma proteins, using two‐dimensional SDS‐PAGE. Animals were weaned at 30 days and kept in feedlots thereafter, receiving hay, concentrate (18% of crude protein) and mineral supplement. Body weight increased from 12.3 ± 0.7 to 54.3 ± 1.6 kg between 8 and 48 weeks (p < 0.05), but changes in thoracic perimeter and scrotal circumference were non‐significant after 36 weeks (p > 0.05). The percentage of motile sperm increased slowly until 23 weeks and more rapidly after that age, but significant changes in progressive motility occurred after 25 weeks. Presence of abnormal sperm related inversely to age. Most significant changes in sperm concentration occurred between 38 and 44 weeks (0.38 ± 0.05 to 1.14 ± 0.24 × 10⁹ cells/ml, p < 0.05) and testosterone reached its highest concentrations at 42 weeks, decreasing afterwards. Rams reached puberty at 28.2 ± 0.8 weeks. The number of protein spots on seminal plasma gels was similar from 15 to 18 weeks (45 and 47 spots; p > 0.05), increased until 24 weeks (141 spots) and 28 weeks (170 spots; p < 0.05) and remained without significant (p > 0.05) changes from 28 to 48 weeks (186 ± 10 spots). Furthermore, the intensity of selected spots on 2D maps increased (p < 0.05) between 15 and 28 weeks, which preceded or coincided with the main developmental changes in sperm motility and percentage of defective sperm in the ejaculates. These results will support future studies designed to characterize specific seminal plasma proteins whose expression relate to the development of testis, epididymis and accessory sex glands.     

Anatomy and Histology of the Male Reproductive Tract and Spermatogenesis Fine Structure in the Lesser Anteater (Tamandua tetradactyla,Myrmecophagidae, Xenarthra): Morphological Evidences of Reproductive Functions

The anatomy and histology of the male genital tract of the lesser anteater were studied. Fine details of spermatozoa regarding their genesis and morphology were also studied in six adult specimens. The testes lie in the pelvic cavity. The deferent duct emerges from the epididymis and opens into the ejaculatory duct, which drains into the membranous urethra. Accessory glands (prostate, seminal vesicle and bulbourethral gland) are histologically similar to those described in other mammals. The short penis presents an urethral orifice, while the corpus spongiosum becomes thinner at the end indicating the absence of a histologically defined glans. The seminiferous epithelium shows: (1) Sertoli cells with deep nuclear indentations, (2) spermatogonia with crusty‐like chromatin, (3) spermatocytes at different stages of maturation and (4) three morphologically distinct stages of spermatid differentiation according to nuclear shape, acrosome development and chromatin condensation. Sperm heads appear oval. The length of the spermatozoa averages 67.33 ± 1.60 μm. Two specimens with inactive spermatogenesis were azoospermic. Their testes and epididymis presented sizes smaller than those with active spermatogenesis. These studies together with others in anteaters may contribute to successful breeding in conservation programmes.     

Effect of weaning method on body weight,testicular growth,sexual behavior,age of puberty,and testosterone concentration of Beetal bucks

The current study evaluates the effects of early (8th week) and late (16th week of age) weaning of male goat kids on their body growth, testicular growth, sexual behavior, plasma testosterone concentration, and pubertal age. Early (n = 6) and late (n = 7) weaned Beetal bucks were weekly monitored from 18th to 38th week for their body weight, scrotal circumference, testicular volume, testicular echogenicity (via ultrasonography), sexual activities, and plasma testosterone concentration. In comparison to early-weaned, late-weaned bucks showed a marked increase (p < .05) in body weight (11.4 ± 0.8 vs. 13.7 ± 0.6kg), testicular volume (44.1 ± 7.2 vs. 79.8 ± 18.7cm³), scrotal circumference (10.7 ± 0.6 vs. 12.8 ± 0.7cm), and testicular echogenicity (28.3 ± 2.7 vs. 38.3 ± 2.1) from 18th, 28th, 21st, and 24th week onward, respectively. Sexual activities started earlier in late- than early-weaned bucks (22nd vs. 25th week, respectively). Moreover, the sexual behavior index was better (p < .05) after the 34th week in late than early-weaned bucks. The plasma concentration of testosterone (at 39 weeks of age) was relatively more and the onset of puberty was 2–3 weeks earlier (p < .05) in late than early-weaned bucks. In conclusion, age-based early weaning of male kids impairs their testicular growth, sexual behavior, and age at puberty compared to conventional weaning.     

Prostasomes of Canine Seminal Plasma – Zinc‐Binding Ability and Effects on Motility Characteristics and Plasma Membrane Integrity of Spermatozoa

Prostasomes are small lipid membrane‐confined vesicles that are involved in various fertilization‐related processes. The aim of this study was to demonstrate canine seminal plasma prostasomes' ability to bind zinc ions, as well as examining their effects on sperm motility characteristics and plasma membrane integrity during cold storage. Ejaculates, collected from five cross‐bred dogs (n = 50), were subjected to ultracentrifugation followed by gel filtration (GF) on a Superose 6 column. Prostasomes appeared as a single fraction in the elution profile. Transmission electron microscopy (TEM) analysis of canine prostasomes revealed the presence of membrane vesicles with diameters ranging from 20.3 to 301 nm. The zinc‐affinity chromatography on a Chelating Sepharose Fast Flow – Zn^{2 +} showed that from 93 to 100% of the prostasome proteins bind zinc ions (P⁺Zn). SDS‐PAGE revealed that canine P⁺Zn comprised four protein bands, with low molecular weights (10.2–12 kDa). We have also shown a positive effect of prostasomes (p < 0.05), especially variant B (2% of total seminal plasma protein) on canine sperm motility parameters after 2 h storage at 5°C (TMOT%, 44.75 ± 5.18) and PMOT%, 12.42 ± 1.59) and VAP, VSL, VCL, when compared with Control (TMOT%, 7.30 ± 1.41 and PMOT%, 1.70 ± 0.42). Higher percentage of spermatozoa with intact plasma membrane (SYBR/PI dual staining) and intact acrosome (Giemsa stained), after 2 h storage at 5°C, was showed, in variant A (1.5% of total seminal plasma protein) and B, when compared with Control and variant C (2.5% of total seminal plasma protein). The prostasomes' effect on motility and plasma membrane integrity of canine cold‐stored spermatozoa may be related to their ability to bind zinc ions and regulate their availability to the sperm.     

Evaluation of a pinhole castration technique in ponies: Comparing single with double ligation (using silk or catgut) of the spermatic cord

The objective of the study was to evaluate a pinhole castration technique in male ponies. Adult ponies (n = 12) were randomly allotted to one of the two equal groups. Both of the groups were anaesthetised with xylazine–ketamine–diazepam and had single (SLS, SLC) ligation of the spermatic cord on one side and double (DLS, DLC) ligation on the other side using silk (Group T1) and catgut (Group T2). Single ligation whether using silk or catgut (SLS and SLC) was completed in 3.50 ± 0.34 min. DLS took 6.66 ± 0.49 min and DLC 7.16 ± 0.47 min. Scrotal oedema was noticed in all of the ponies from Day 1 to Day 15. The scrotal circumference and testicular volume in animals of both of the groups showed a significant (P<0.05) increase from Day 1 to Day 4. Orchiectomy to recover testicular remnants was performed in all of the animals on Day 41. Straw-coloured fluid accumulation was noticed in seven testicular remnants (4 T1 and 3 T2). One testicle from Group DLS showed suppuration. Multiple gross and histological abnormalities were detected in all double ligated testes. The changes were more severe in the DLS than the DLC group and their epididymis (n = 3) also showed necrosis, fibroplasia and an obstructed lumen. Severe adhesions had developed in three and one testicle only from the SLS and SLC groups respectively. The remaining testes and epididymis in both of these groups showed only mild-to-moderate adhesions. From this study, it was concluded that castration may not be achieved by percutaneous single ligation of the spermatic cord in ponies. Although double ligation induces marked gross and histopathological changes, assessments of the testosterone levels and sperm analysis are required before recommending this procedure. Use of silk for ligation of the spermatic cord is advantageous over catgut but maintenance of strict asepsis is mandatory.     

Comparison of two methods of seminal plasma removal on buffalo (Bubalus bubalis) sperm cryopreservation

Cryopreservation causes damage to spermatozoa, and methods minimizing this damage are therefore needed. Although much discussed, seminal plasma removal has become an alternative to improve sperm quality and viability after freezing and has been applied to different species in attempt to obtain good results. The objective of this study was to evaluate semen quality in buffaloes submitted to two methods for seminal plasma removal (filtration and centrifugation). Semen samples were collected from seven Murrah buffalo bulls (Bubalus bubalis) once a week for 8 weeks. Each ejaculate was divided into three groups: control (presence of seminal plasma), centrifugation and filtration. Sperm kinetics was evaluated with the computer‐assisted sperm analysis (CASA) system. Plasmalemma and acrosomal membrane integrity, mitochondrial membrane potential and reactive oxygen species (ROS) were measured by flow cytometry, and lipid peroxidation was evaluated by the thiobarbituric acid reactive substances (TBARS) assay. Seminal plasma removal did not improve sperm kinetics compared to the control group. Centrifugation increased the number of cells with damaged acrosomal membranes (0.77 ± 0.05) and filtration caused greater plasmalemma and acrosomal membrane damage (22.18 ± 1.07). No difference in the mitochondrial membrane potential was observed between groups. In contrast, ROS production was higher in the centrifugation group compared to the control and filtration groups, although no differences in TBARS formation were detected. In conclusion, seminal plasma removal did not improve the quality of thawed buffalo semen compared to control in terms of sperm kinetics, membrane integrity, mitochondrial membrane potential or lipid peroxidation.     

Correlation of testicular ultrasonography,testicular biometry,serum testosterone levels and seminal attributes in pre and post-pubertal age for breeding soundness evaluation in Osmanabadi bucks

The present research work entitled “Correlation of testicular ultrasonography, testicular biometry, serum testosterone levels and seminal attributes in pre- and post-pubertal age for breeding soundness evaluation in Osmanabadi bucks” was undertaken in 18 healthy Osmanabadi bucks from the Instructional Livestock Farm Complex, Bombay Veterinary College, Mumbai, Maharashtra. The body weight (kg), scrotal circumference (cm) and testicular biometry (cm) of post-weaning 18 Osmanabadi male kids was recorded every 15 days from weaning, i.e., 120?±?10 days along with serum testosterone (ng/ml) by radioimmunoassay method at monthly intervals for the next 6 months. Semen was collected six times on the seventh month onward during post-pubertal age at 15-day interval from 18 bucks. The semen was evaluated for macroscopic and microscopic tests. The body weight increased from 14.45?±?0.67 to 19.57?±?0.70 kg from four to nine and a half months of age. The average daily body weight gain was 31.27 g. Maximum body weight gain was 01.19?±?0.16 kg from 5 to 6 followed by 01.15?±?0.16 kg from 4 to 5 months of age. The scrotal circumference increased from 17.22?±?0.56 to 19.03?±?0.55 cm from four to nine and a half months of age with maximum increased between 4 and 5 followed by 6 and 7 months of age. The testicular length, width and thickness of right and left testicles were recorded by ultrasonography method. There was increase in mean right and left testicular length, width and thickness from 5.25?±?0.19 to 5.84?±?0.18 and 5.49?±?0.21 to 6.16?±?0.20; 2.99?±?0.12 to 3.32?±?0.12 and 3.10?±?0.13 to 3.44?±?0.12 and 2.97?±?0.12 to 3.16?±?0.12 and 3.06?±?0.12 to 3.31?±?0.11 cm, respectively by ultrasonography, between four to nine and a half months of age. Testicular length, width and thickness gain was at maximum in 5 to 6 months of age. Left testicular length was more than the right testis. Before puberty, there was sudden gain in body weight, testicular length and width. However, scrotal circumference showed significant increase after puberty. Body weight had highest correlation with ultrasonographic left testicular thickness (r?=?1) followed by scrotal circumference, ultrasonographic right and left testicular width, left testicular length, right testicular length and thickness and least by right testicular thickness (r?=?0.95). The semen was thin to thick in consistency and average semen density was 3.10?±?0.05. Average semen volume was 0.81?±?0.02 ml, mass activity, initial motility, live and dead sperm count, abnormal sperm count and sperm concentration were 3.45?±?0.13, 76.16?±?1.16 and 75.16?±?1.28% and 24.84?±?1.28, 12.30?±?0.50% and 2631.04?±?45.74 million/ml, respectively in 18 bucks in six collection at 15 days. There was significant rise in semen volume, mass activity, initial motility and concentration at 8.5 months and live count, density at 9 months of age which indicates the age of sexual maturity is 8.5 to 9 months in Osmanabadi bucks. The body weight had highest positive correlation with mass activity (r?=?98) followed by initial motility, live sperm count and total sperm concentration, semen volume (r?=?76). The scrotal circumference had highest positive correlation with initial motility (r?=?98) followed by live sperm count, total sperm count, mass activity, semen volume (r?=?86). On the other hand, body weight and scrotal circumference were negatively correlated with abnormal and dead sperm count. The mean testosterone concentration increased from 0.02?±?0.004 to 5.75?±?0.80 ng/ml between four and half to nine and half months of age, respectively. There was significant rise (p?<?0.01) up to 1.38?±?0.28 ng/ ml at 6.5 months, i.e., age of puberty and up to 5.75?±?0.80 ng/ml at 9.5 months, i.e., age of sexual maturity. Testosterone had highest positive correlation with testicular length followed by testicular width, length, body weight and scrotal circumference, mass activity, live sperm count, initial motility, while it had highest negative correlation with dead and abnormal sperm count. From the present research work, it was concluded that the scrotal circumference, testicular length, width and thickness increased with increasing body weight. Before puberty, there was sudden gain in body weight, testicular length and width. However, scrotal circumference increased significantly at post-pubertal age. So testicular length, body weight, testicular width in pre pubertal age and scrotal circumference post-pubertal age can be used as indicator for selection of Osmanabadi bucks for breeding purpose. On the other hand, the semen parameters should consider only after 8.5 to 9 months of age for selection of Osmanabadi bucks for breeding.

     

Changes in testicular size,echotexture, and arterial blood flow associated with the attainment of puberty in Dorper rams raised in a subtropical climate

There is a paucity of information on the relationships of testicular morphology, echotextural attributes, and blood flow dynamics with pubertal development of rams raised in a subtropical climate. Forty‐five Dorper rams (24 rams aged 8–11 months and 21 rams aged 12–24 months) were examined using a portable ultrasound scanner connected to a 7.5‐MHz transducer. Computer‐assisted analyses of testicular ultrasonograms utilized commercially available Image ProPlus^® analytical software. Spectral Doppler scans of testicular arteries were performed immediately after scrotal (B‐mode) ultrasonography to determine peak systolic velocity (PSV), end‐diastolic velocity (EDV), resistive index (RI = [PSV?EDV]/PSV), and pulsatility index (PI = [SPV–EDV]/mean velocity) of the blood vessels. The length of the testes (9.7 ± 0.3 compared with 9.0 ± 0.2 cm) and scrotal circumference (33.3 ± 0.5 compared with 31.8 ± 0.4 cm) were greater (p < 0.05) but testicular depth (4.5 ± 0.1  compared with 4.9 ± 0.08 cm) was less (p < 0.05) in sexually mature compared with peripubertal rams. [Corrections added on 9 Jan 2019 after initial online publication: The testicular size values in the sentence were corrected.] There were no differences (p > 0.05) between the two age groups of Dorper rams in blood flow indices of testicular arteries. Mean numerical pixel values (100.5 ± 4.1 compared with 89.2 ± 4.8) and pixel heterogeneity (25.6 ± 0.6 compared with 23.6 ± 0.5) of testicular parenchyma were greater (p < 0.05) in peripubertal than in postpubertal rams. Semen volume was negatively correlated with PI of testicular arteries (r = ?0.57, p = 0.04). In summary, the attainment of sexual maturity in the rams of the present study was associated with significant changes in testicular length and depth, scrotal circumference, and parenchymal echogenicity/hetrogeneity but not in testicular volume and blood perfusion rates. Testicular artery PI can be used to predict the volume of ejaculate in rams.     

Retracted: Study of enzyme activities and protein content of beluga (Huso huso) semen before and after cryopreservation

Knowledge gained regarding the biochemical processes that occur during sperm collection, processing and freezing‐thawing might improve current sperm cryopreservation techniques. In our present study, we determined the effect of cryopreservation on the total protein concentration (TP) and the activities of certain enzymes in semen samples from the beluga (Huso huso). The TP content of the seminal plasma of fresh semen was 0.47 ± 0.026 g/l, and the TP after cryopreservation was 1.86 ± 0.6 g/l. The activities of acid phosphatase (0.82 ± 0.042 U/l), lactate dehydrogenase (234.4 ± 19.4 U/l), arylsulfatase (143.1 ± 32.5 U/l) and β‐N‐acetylglucosaminidase (58.39 ± 4.14 U/l) in the seminal plasma of fresh semen were significantly lower than those in the supernatant of frozen‐thawed semen samples (7.43 ± 0.64, 3224.6 ± 167.2, 422.6 ± 21.3 and 90.2 ± 5.37 U/l respectively). These parameters may be useful as biomarkers for estimating damage to the cell membrane of spermatozoa caused by freezing‐thawing.     

Factors associated with low vitamin D status of Australian alpacas

Objective To investigate factors associated with low vitamin D status of alpacas at pasture in southern Australia. Design A 2‐year survey of alpacas from two farms in South Australia and three in Victoria. Blood samples were collected from 20 to 30 alpacas on each farm on five occasions each year. Breed, gender, age and fleece colour of animals were recorded. Method Blood samples were assayed for plasma 2.5‐hydroxycholecalciferol (25‐OH D₃) and plasma inorganic phosphorus (Pi). Data sets from 802 animal samples were analysed by multiple regression to determine variables associated with low vitamin D status of alpacas. The relationship between plasma 25‐OH D₃ and plasma Pi was also investigated. Results Vitamin D status was significantly affected by month of sampling, with low values in late winter and high values in summer. Plasma vitamin D concentrations increased with age, were higher in alpacas with light fleeces than in those with dark fleeces and were also higher in the Suri than in the Huacaya breed. Plasma Pi concentrations were generally lower in alpacas with plasma 25‐OH D₃ values < 25 nmol/L. Conclusions Young alpacas with dark fleeces are most at risk from vitamin D insufficiency in late winter in southern Australia. The present study indicates that plasma Pi values are not a reliable indicator of vitamin D status of alpacas as assessed by plasma 25‐OH D₃ concentrations.     

Clinical,Ultrasonographic and Pathological Findings in a Bull with Segmental Aplasia of the Mesonephric Duct

On assessment for use in an AI stud, a 12‐month‐old bull was found to produce low volume ejaculates with 41% of the sperm having morphological abnormalities. No left epididymal tail was palpable and the head of the epididymis on the left was twice the size compared with the right. Ultrasound examination showed the left testis to contain a large central area of decreased echogenicity, which could be followed proximally to a 15‐mm echolucent lesion at the site of the epididymal head. Postmortem examination revealed a 15‐mm diameter cyst in the region of the left epididymal head, and absence of the body and tail of the epididymis. The mediastinum testis of the left testis was dilated, corresponding to the area of decreased echogenicity observed on ultrasonography. No left seminal vesicle was present and the ampulla was significantly smaller than the same structure on the right. Histological examination revealed incomplete or absent spermatogenesis involving the majority of seminiferous tubules in the left testis, and a small proportion of those of the right testis. The cystic structure at the site of the left epididymal head was lined by irregular, sometimes attenuated, epithelium and contained sparse spermatozoa. This case demonstrates the adverse impact, which segmental aplasia of the mesonephric duct had on the testicular and epididymal function of a bull, and highlights the importance of careful clinical assessment in its diagnosis.     

Paraoxonase 1 (PON1) activity in serum,follicular fluid and seminal plasma of sheep

This work aimed to describe the activity of paraoxonase 1 (PON1) in serum, follicular fluid and seminal plasma of sheep. Average serum PON1 activity was 286.8 ± 96.2 U/ml in females and 237.6 ± 18.9 U/ml in males. There was a positive correlation between PON1 activity in serum and follicular fluid in females, being twice higher in serum than in follicular fluid (148.8 ± 15.7 U/ml). PON1 activity in males’ serum was 10‐fold higher than in seminal plasma (21.18 ± 14.2 U/ml), and there was no correlation between PON1 activity in both compartments. Finally, this work suggests that PON1 activity of in sheep is higher compared to other mammalian species, and there is an association between PON1 in serum and follicular fluid only.