Effects of Sperm Concentration and Straw Volume on Motion Characteristics and Plasma, Acrosomal, and Mitochondrial Membranes of Equine Cryopreserved Spermatozoa

The advantages of using cryopreserved semen in equine reproduction are well known. During cryopreservation, spermatozoa undergo many changes that lead to a decrease in fertility. There is no agreement on the ideal sperm dose and concentration to maximize fertility rates. Thus, the objectives of this experiment were to evaluate sperm motion by computer-assisted analysis (CASA), sperm membrane integrity and function with fluorescence probes of cryopreserved sperm at three concentrations: 100 (C100), 200 (C200) and 400 × 10⁶ sperm/mL (C400), and two straw volumes (0.50 and 0.25 mL). There was no interaction between sperm concentration and storage volume (P > .05). Sperm motion characteristics were influenced by concentration (C100 > C200 > C400; P < .05). Curvilinear velocity (VCL) in 0.25-mL straws had higher average values (P < .05). Membrane integrity and function were not changed by straw volume (P > .05). However, sperm concentration changed the percentage of cells with intact plasma membrane (C100 > C200 > C400; P < .05) and the percentage of cells with high mitochondrial membrane potential (C100 = C200; P > .05 and C400 < C100 and C200; P < .05). According to this experiment, the best freezing method was that involving 100 × 10⁶ sperm/mL, regardless of straw volume.     

Preliminary study of ovarian activity in fillies treated with a GnRH vaccine

Objective To investigate the effects of two doses (200 and 400 mg) of a water-soluble gonadotrophin-releasing hormone vaccine on the ovarian activity of 2-year-old fillies.
Design A controlled vaccination dose rate experiment.
Animals Six 2-year-old Australian Stock Horse fillies were randomly allocated to three treatment groups: unvaccinated controls, those receiving 200 mg of the vaccine and those receiving 400 mg of the vaccine.
Results Ovarian activity of the treated fillies was suppressed at the peak of the breeding season while that of the untreated controls continued normally. The control fillies displayed oestrous activity and behaviour. Suppression of ovarian activity occurred for 25 and 30 weeks in the 200 and 400 mg groups, respectively. These differences were not significant. Ovarian activity ceased 2 to 3 weeks after primary vaccination. Antibody titres were low (330) until after the booster immunisation when they rapidly peaked at 22,000 and 28,000 in the 200 mg and 400 mg groups, respectively. Plasma progesterone concentrations of the treated fillies remained below 3.18 nmol/L while GnRH was suppressed. The vaccine had no significant effect on plasma androstenedione concentrations. Recovery from the effect of the vaccine was associated with development of ovarian follicles, normal oestrous behaviour and ovulation. Three of the four treated fillies and one of the controls conceived during the next breeding season and foaled normally. All the treated fillies conceived and produced normal foals in the following two breeding seasons.
Conclusion Both dose rates suppressed ovarian function and prevented oestrous behaviour. These effects were reversible and the subsequent fertility of the vaccinated fillies was normal.     

Effect of active immunization against GnRH on testosterone concentration,libido and sperm quality in mature AI boars

Background

The aim of the present study was to investigate the efficacy of the Improvac on testosterone concentration in blood serum, sexual behavior and sperm quality in matured AI boars. A total of nine Danish Landrace AI boars were included in the analysis.

Methods

The trial period lasted for 15 weeks and was divided into four periods: Control period: three weeks before vaccination; Period I – four weeks after first vaccination; Period II – four weeks after second vaccination, Period III – four weeks after third vaccination. Blood and sperm samples were collected at weekly intervals. Freshly collected sperm samples were analyzed.

Results

Testosterone concentration correlated with libido (r = 0.531; p < 0.001), volume of ejaculate (r = 0.324; p < 0.001) and the percentage of morphologically normal spermatozoa (r = 0.207; p < 0.05). Testosterone concentration rised significantly (p < 0.05) in 5–6 week of trial, e. i. after the first dose of Improvac and after this peak the level of testosterone further progressively decreased (p < 0.05).

Conclusions

Results from this study indicate that active immunization of sexually matured boars against GnRH has negative impact on testosterone concentration, sexual behavior, volume of ejaculate and total number of normal spermatozoa in ejaculate.     

Effects of Intratesticular Injection of 70% Glycerin on Stallions

The removal of endogenous germ cells of recipient stallions is a key step to produce donor germ cell-derived sperm using the germ cell transplantation technique. Six Thoroughbred stallions were divided into a treatment (n = 3) and a control group (n = 3), and 70% glycerin (1, 2, 3-trihydroxypropane, 40 mL per testis) or phosphate-buffered saline, respectively, was locally injected into testes. General semen evaluation, libido, and testicular volume were performed weekly from 3 weeks before to 10 weeks after treatment. The number of round germ cells in the ejaculate was counted using a hemocytometer. The hematoxylin and eosin staining was performed on the cross sections of testicular tissue obtained 11th week of treatment. Plasma testosterone levels in blood collected weekly were measured using a colorimetric competitive enzyme immunoassay kit. The sperm number was significantly lower than that of the control group at 5 and 10 weeks after glycerin injection. No differences in the status of spermatogenesis in the cross sections of seminiferous tubules and testicular volume were found between the two groups. The 70% glycerin-treated stallions had reduced total and progressively motile sperm and exhibited a significantly higher population of round germ cells in the ejaculate. Testosterone levels, testicular volumes, and libido of stallions were not significantly different between the groups. In conclusion, although intratesticular injection of 70% glycerin may have caused disassociation of some germ cells in the seminiferous tubules for several weeks, it did not significantly ablate germ cells in the tubules at 11 week in stallions.     

Changes in Testicular Steroidogenic Acute Regulatory (STAR) Protein,Steroidogenic Enzymes and Testicular Morphology Associated with Increased Testosterone Secretion in Bulls Receiving The Luteinizing Hormone Releasing Hormone Agonist Deslorelin

Testosterone secretion and the expression and relative contents of steroidogenic acute regulatory (StAR) protein and steroidogenic enzymes cholesterol side-chain cleavage cytochrome P450 (P450_scc), 3β-hydroxysteroid dehydrogenase /Δ⁵ → Δ⁴ − isomerase (3β-HSD), and 17α-hydroxylase cytochrome P450/C_17–20 lyase (P450_17α) were determined in testicular tissues of bulls treated with a LHRH agonist. Testis morphology and spermatogenesis were also examined. In Experiment 1, bulls (30-mo-old) received no treatment (control, n = 7) or were implanted for 10 days with the LHRH agonist deslorelin (n = 7). Bulls were castrated on Day 10 and testis tissues prepared for Western and Northern blotting. At castration, bulls implanted with deslorelin had greater plasma testosterone (5-fold) and testis content of testosterone (10-fold) compared with control bulls. Relative content (per μg total testis protein or RNA) of StAR protein, 3β-HSD, P450_scc, and mRNA for P450_17α in bulls treated with deslorelin ranged from 3- to 6-fold that of control bulls. In Experiment 2, bulls (20-mo-old) were left untreated (control, n = 6) or implanted with deslorelin (n = 12) for 120 days. On Day 120, bulls were castrated and right testis tissues prepared for morphology. Testis volume and weight were increased (P < 0.01) in bulls treated with deslorelin compared with control bulls. Stereological analysis revealed that this increase occurred in all compartments (seminiferous epithelium, lumen and interstitium) studied, but was significant (P < 0.01) only for the seminiferous epithelium. Absolute numbers of round spermatids per testis were increased (P < 0.05) in bulls treated with deslorelin compared with control bulls. Increased testosterone secretion in bulls treated with deslorelin was associated with increased testicular StAR protein and steroidogenic enzymes. Bulls treated long-term with deslorelin had a faster rate of testis growth and increased daily sperm production at the end of the experiment.     

大肠杆菌油佐剂灭活苗免疫雏鸡抗体消长规律的研究

本试验用鸡源致病性大肠埃希氏菌Ｏ２血清型制成的油佐剂灭活苗对１５日龄雏鸡进行免疫接种，以测定其抗体消长规律；对免疫后不同抗体滴度的雏鸡进行了攻毒试验．结果表明，免疫１周后可监出抗体，２～３周抗体达到高峰，抗体可持续１０周左右．抵抗同型大肠杆菌攻霉的最低抗体滴度为１：１６～１：３．免疫保护期可达２个目以上．     

THE SEROLOGICAL RESPONSE OF CHICKENS TO AUSTRALIAN LENTOGENIC STRAINS OF NEWCASTLE DISEASE VIRUS

SUMMARY: Australian lentogenic Newcastle disease viruses were evaluated as uninactivated vaccines in Australian chickens, the response being evaluated by the production of haemagglutination-inhibition (HI) antibodies. Two viruses, V4 and PM9, induced high levels of antibody and were readily transmissible between chickens by contact exposure. Three other viruses were poorly immunogenic and poorly transmissible. Chickens vaccinated intramuscularly with the V4 strain produced higher HI antibody titres than chickens vaccinated by the orotracheal, intranasal and intraocular routes. HI antibody titres in chickens vaccinated with the V4 strain reached peak levels 3 to 5 weeks after vaccination and waned considerably during the next 2 to 4 weeks. However, low levels of HI antibody persisted for at least 36 weeks after vaccination. Intramuscular vaccination with the V4 strain of one-day-old chicks lacking maternal antibody to Newcastle disease virus resulted in 42–70% mortality and the survivors developed very high titres of HI antibody. Similar chickens inoculated orotracheally showed signs of depression and developed high titres of HI antibody, but there were no mortalities. Chickens 1-, 2-, 3- and 4-weeks-old and lacking maternally derived HI antibody to Newcastle disease virus suffered no adverse reaction to intramuscular or orotracheal vaccination. The antibody response of the 1-week-old chickens was considerably poorer than that of the older chickens. Following orotracheal vaccination with the V4 strain, chickens with low levels of maternally derived antibody responded with low levels of HI antibody. On the other hand, in the progeny of hens hyperimmunised with the V4 strain the production of active antibody following orotracheal vaccination was delayed until the level of passive antibody had declined considerably. There was no response to intramuscular vaccination in congenitally hyperimmune chickens. The minimum HI antibody inducing dose of V4 vaccine, when measured 3 weeks after vaccination of 6-weeks-old chickens, was 10^5.6 50% egg infectious doses.     

Preliminary testing in turkeys of the safety and efficacy of a putative haemorrhagic enteritis virus vaccine

Haemorrhagic enteritis virus (HEV) causes clinical haemorrhagic enteritis in young poults and/or subclinical immunosuppression which is often associated with colibacillosis. This disease is controlled with live vaccines worldwide, however, importation of HEV vaccines or cells that support HEV propagation are not permitted in Australia. A major experiment in isolators was conducted to test the safety and efficacy of a putative HEV vaccine. The study had a factorial design with four factors namely vaccination age (28 and 42 days of age), vaccine dose (0, 10⁵, 10⁶, 10⁷ genomic copies of HEV vaccine), challenge with HEV (yes, no) and vaccination‐challenge interval (7, 21 or 42 days). A total of 315 poults were used providing 6‐8 birds per treatment combination. Turkey growth rate, mortality, pathological findings, anti‐HEV antibodies and viral load were examined. Vaccination lead to significant increases in anti HEV antibody over the following 2‐4 weeks. Overall, vaccination with 10⁶ and 10⁷ was protective against increase in relative splenic weight and splenic viral load in challenged birds. Clinical haemorrhagic enteritis was not induced by any treatment but there was an increased incidence of airsacculitis in groups receiving either HEV vaccine or challenge virus compared to the negative control birds (25.8‐29.3% vs. 9.4%, P < 0.05). Growth rate, mortality and relative bursal weight were unaffected by vaccination. This laboratory level study indicates that the putative vaccine is safe and likely to be efficacious, but may cause elevated levels of airsacculitis. These findings require confirmation in larger scale field trials.     

Flow cytometric assessment of ethylene glycol monoethyl ether on spermatogenesis in rats

The effects of ethylene glycol monoethyl ether (EGEE) on testicular cell populations in rats were investigated by a flow cytometric method. Rats were administered by gavage with EGEE at the various doses of 0 (saline alone), 100, 200, 400, and 800 mg/kg body weight/day for 4 weeks. The treatment of EGEE caused decreases in the weight of testis and epididymis and in the number of testicular cells. Histopathologically, exfoliation of germ cells into the tubular lumen was observed at the doses of above 200 mg/kg. The treatment of EGEE at the dose of 400 mg/kg caused moderate testicular degeneration. A significant depletion of haploid cells and a disproportionate ratio of diploid and tetraploid cells were observed as determined by flow cytometric analysis. These results indicate that the toxic effect of EGEE on the male reproductive system may be strongly associated with the disproportion of testicular germ cells.     

Function of contralateral testis after artificial unilateral cryptorchidism in dogs

The effects of a cryptorchid testis on the contralateral testis were investigated after artificially producing unilateral cryptorchidism in 8 beagle dogs. Bilateral testicular biopsy and collection of spermatic vein blood and peripheral vein blood were performed at the time of the operation to produce the cryptorchidism and 52 weeks later. The testicular tissue was used for histological examination by light microscopy and measurement of the testicular transferrin (Tf) concentration by enzyme immunoassay. Plasma testosterone (T), estradiol-17 beta (E2), and luteinizing hormone (LH) levels were measured by radioimmunoassay. Semen was collected weekly and its quality was examined. No spermatogenesis was observed in the cryptorchid testes at 52 weeks after the operation, and the number of germ cells in the contralateral testes had decreased but the number of Sertoli cells did not change. The Tf concentration in both testes had also decreased. The mean total number of sperm between 48 and 52 weeks after the operation (194 x 10(6)) was less than half the number before the operation (510 x 10(6)). Mean spermatic vein plasma T levels (51 ng/ml) in the cryptorchid testes 52 weeks after the cryptorchid operation were significantly lower than before the operation (91 ng/ml; P < 0.05). By contrast, spermatic vein plasma E2 levels (80 pg/ml) were significantly higher than the values before the operation (51 pg/ml P < 0.05). The peripheral plasma LH levels decreased. These findings indicate that a large quantity of E2 secreted by the cryptorchid testis inhibits the endocrine and spermatogenic functions of the contralateral testis in the dog. In particular, it is assumed that dysfunction of the contralateral testis is associated with Sertoli cell dysfunction suggested by the low Tf concentration.     

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.

     

An evaluation of the anamnestic test for brucellosis in cattle of the northern pastoral area

SUMMARY An investigation of the anamnestic test for brucellosis using Brucella abortus 45/20 vaccine was carried out in 3 groups of weaner cattle on 2 farms in western Queensland. Each group originally consisted of about 500 cattle. They were bled before and at 6 or 10 weeks after vaccination and again in the following year. The serums were tested by the complement fixation (CFT), Rose Bengal (RBT) and indirect haemolysis tests (IHLT). Most of the cattle reacting to one or more of the tests were killed and selected tissues were subjected to bacteriological examination for B. abortus. B. abortus was isolated from 19 of 30 (63%) pre-vaccinal reactors, 23 (24%) of 96 cattle reacting at 6 or 10 weeks after vaccination (the anamnestic test) and 1 (2%) of 50 cattle reacting one year after vaccination. The reactor found to be infected the year after vaccination had high serological titres in each of the 3 serological tests: RBT of 3, CFT of 128 and IHLT of 256. A subsequent test showed the group to be brucellosis-free. The CFT was the most efficient test. In the pre-vaccination tests 17 of 19 infected animals were positive in the CFT compared with 11 positive in the IHLT and 17 in the RBT. In post vaccination tests 22 of 23 infected animals were positive in the CFT compared with 18 in the IHLT and 19 in the RBT. At the pre-vaccinal and anamnestic tests (6 or 10 weeks after vaccination) 19 of 57 (33%) cattle with CF titre of 4 or 8 yielded B. abortus on culture compared with none of 26 cattle with similar titres in the year after vaccination. The interpretation of CF titres in cattle following 45/20 vaccination needs to be re-examined.     

Testicular Measurements and Daily Sperm Output of Tori and Estonian Breed Stallions

Evaluation of testicular measurements and daily sperm output (DSO) yields valuable information for predicting the reproductive capacity of stallions. The present study evaluated testicular measurements (height, length, width and circumference) and DSO of eight Tori and eight Estonian breed stallions. One ejaculate of semen was collected daily for 10 subsequent days from each stallion. The gel‐free volume of semen was measured with a graduated glass cylinder and the sperm concentration was assessed with a Chorjajev chamber. The volume of gel‐free fraction was multiplied by the sperm concentration to give the total number of spermatozoa (TSN). The DSO was calculated as mean TSN of collection on days 8–10 in Tori breed stallions and on days 4–10 in Estonian breed stallions. The DSO of Tori breed stallions was 12.9 × 10⁹ spermatozoa and of Estonian breed stallions 4.5 × 10⁹ spermatozoa (p < 0.001). Testicular measurements (in cm) 1 day after the last semen collection were as follows: left testis– height 7.3, length 10.4 and width 7.3 in Tori breed stallions, and 5.9, 8.1 and 5.9, respectively, in Estonian breed stallions; right testis– height 7.4, length 10.6 and width 7.4 in Tori breed stallions, and 5.5, 7.4 and 5.3, respectively, in Estonian breed stallions. All these testicular measurements were significantly smaller in Estonian than in Tori breed stallions (p < 0.001). Testicular circumference was 45.4 and 35.4 cm in Tori and Estonian breed stallions, respectively (p < 0.001). The testicular circumference was correlated with DSO in both Estonian (p < 0.05) and Tori breed stallions (p = 0.071). The results give us valuable information on the reproductive capacity of Tori and Estonian breed stallions.     

Therapeutic Ultrasound as a Potential Male Dog Contraceptive: Determination of the Most Effective Application Protocol

Ultrasound is one of the most promising forms of non‐invasive contraception and has been studied in several animal models. The objective of the current investigation was to determine the most practical and effective application protocol for dog sterilization. A total of 100 dogs were divided into five equal groups. Group A received 5‐min applications three times performed at 48‐hr intervals and covering the entire testicular area at frequency of 1 MHz; Group B received 5‐min applications three times performed at 48‐hr intervals over the dorso‐cranial area of the testis at frequency of 3 MHz; Group C received three sequential 5‐min applications (at 5‐min intervals between applications) covering the entire testicular area at frequency of 1 MHz; Group D received 15‐min applications two times performed at 48‐hr intervals and covering the entire testicular area at frequency of 1 MHz. The experimental groups' ultrasound had an intensity of 1.5W/cm². The Control Group had the same procedure as Group A, but with the transducer switched‐off. Dogs were surgically castrated 40 days following the treatment for histological examination. Azoospermia, testicular volume reduction and apparently irreversible testicular damage were achieved by Group A. No effects were noticed in the other groups. Testosterone levels remained within physiological range with all application protocols. A regimen of three applications of ultrasound at 1 MHz, and 1.5 W/cm², lasting 5 min with an interval of 48 h was effective as permanent sterilization in the dog without hormonal impact.     

Changes in Testicular Development, Ultrasonographic and Histological Appearance of the Testis in Buck Kids Immunized Against LHRH Using Recombinant LHRH Fusion Protein

This study was designed to evaluate the effectiveness of recombinant Ovalbumin-LHRL (OL) immunization on changes in testicular size, histological appearance and testosterone production in buck kids. Thirty native buck kids at 18 weeks of age were divided into three groups, control (n = 10), immunization (n = 10) and castration (n = 10) groups. Immunized animals received OL protein generated by recombinant DNA technology. Ultrasonographic and histological examinations of the testes were performed. Animals were slaughtered at 44 weeks of age. Semen and epididymides were evaluated for the presence of sperm cells. Immunized animals generated anti-LHRH antibodies. Testosterone production, testicular and accessory glands development and sperm production were suppressed in the immunized animals (p < 0.01). Semineferous tubule diameters decreased (p < 0.01), basal membrane of the tubule was thickened and hyalinized in immunized kids. Immunization affected ultrasonographic appearance of the testes drastically. While testes of control animals gained their normal ultrasonographic appearance as the age increased, immunized animals had uniform hypoechogenic testicular structure as observed at 18 weeks of age until slaughter. Simultaneous histological and ultrasonographic evaluations indicated that the changes in testicular histology could partly be monitored via ultrasonographic imaging; nevertheless, it is difficult to claim that ultrasonographic image reflects the exact changes in such instances. In conclusion, these results indicate that recombinant OL fusion protein is effective in immunocastration in buck kids and has a potential to be used as an alternative to physical castration. Further researches should be conducted to help assessing reproductive status of testes from ultrasound images.     

VACCINATION OF HEIFERS WITH A REDUCED DOSE OF BRUCELLA ABORTUS STRAIN 19 VACCINE BEFORE FIRST MATING

Ten Jersey heifers aged 14 to 23 months were vaccinated with 2.25 times 10⁸ cells of living Brucella abortus strain 19 vaccine. They and 10 similar non-vaccinated heifers were subsequently mated and when about 6 months pregnant were challenged by the conjunctival application of a virulent culture of B. abortus. The serological response to vaccination was much less than is usually seen following vaccination with the normal dose of strain 19, especially when the indirect haemolysis test was used. A persistent vaccinal reaction was observed in one heifer. Significant resistance to infection was demonstrated which was greater than that previously observed in calfhood vaccinates given the full dose but less than that shown by cows given the smaller dose in early pregnancy. The effectiveness of strain 19 vaccination appears to be related to the age of the animal at vaccination.     

Fertility of Mares Inseminated With Frozen-Thawed Semen Processed by Single Layer Centrifugation Through a Colloid

The aim of this study was to determine whether there was an increase in pregnancy rates when frozen-thawed stallion semen was processed by single layer centrifugation (SLC) through a colloid before insemination. In addition, changes in semen parameters, including motility, were determined before and after SLC. Twenty light-horse mares (aged 3-16 years) and one Thoroughbred stallion (aged 16 years) having average fertility with fresh and cooled semen (>50% per cycle) and displaying a postthaw motility of >35% were used. Control mares were inseminated using 4- × 0.5-mL straws (200 × 10⁶/mL) of frozen-thawed semen. Treatment mares were inseminated with 4 × 0.5 mL of frozen-thawed semen after processing by SLC. Pregnancy rates were compared using Fisher exact test, and continuous parameters were evaluated by a Student t test. The pregnancy rates at day 14 were not different for the mares inseminated with control versus SLC-processed semen, despite the difference in sperm number (171 × 10⁶ ± 21, 59 × 10⁶ ± 25 progressively motile sperm). After frozen-thawed semen was processed by SLC, the percentage progressively motile sperm improved (P < .05), and SLC processing resulted in a 21.8% recovery of spermatozoa. In summary, centrifugation of frozen-thawed semen through a single layer of colloid increased the percentage of motile spermatozoa, but did not improve pregnancy rates after deep horn insemination.     

Relationship between angioarchitecture of the testicular artery and spermiogram parameters in Egyptian buffalo bulls (bubalus bubalis)

The current study aimed to investigate the effect of testicular artery angioarchitecture on the spermiogram parameters in Egyptian Buffalo bulls. Eight adult buffalo bulls aged between 2 and 8 years were used for semen evaluation. For anatomical studies, the masculine gonads were collected after slaughtering 30 adult bulls and prepared for injection by different masses (Urographine^®, Latex and Epoxy) through the testicular artery. The mass activity of the ejaculate was assessed immediately after collection. The sperm motility in fresh bull ejaculate was more than 80%. The overall mean percentage of sperm abnormalities was <18%. The recorded sperm abnormalities were mostly secondary one including distal protoplasmic droplet, fragmented tail, detached head, detached galea capitis and bent tail. The highest percentage of sperm viability was recorded just after sperm collection (alive > 85%). The results revealed that testicular artery can be divided into three parts (abdominal, funicular and marginal parts) along its course. The coils of the funicular part forming a cone-like structure with its base fixed to the head of the testis. Two epididymal branches to the head and tail of epididymis emanate from the funicular part which continues as pars marginalis on the lateral surface of testis before its division into the lateral and medial testicular arteries on approaching the tail extremity of the testis. The increase in the length of the testicular artery with increase in the size of the testes played a great role in the degree of complexity of the architectural vascular patterns. The degree of complexity is affected by the number of coils formed by the vessel. The increase in the convolutions of the vessel will reduce the speed of blood flow to the gonads. Thus in turn will enabling the thermoregulatory mechanism to work more efficiently and will affect the semen value.     

Effects of the GnRH antagonist acyline on the testis of the domestic cat (Felis catus)

The aim of this study was to describe the effects of a single dose of the gonadotrophin releasing hormone (GnRH) antagonist acyline on testicular characteristics of the domestic cat. Twelve mature cats were orchidectomised unilaterally (right testis) on Day -7 (n=7) or Day 15 (n=5). On Day 0, 330 μg/kg acyline was administered s.c. to all the animals. Left orchidectomy was carried out on Day 15 (n=2), Day 30 (n=4) and Day 60 (n=6). Sperm were recovered from the epididymis and the testes were evaluated grossly, histologically and immunohistochemically. Significant differences (P<0.05) were found between days for epididymal sperm motility, vigor, abnormal morphology, germinal epithelium height, spermatocytes, spermatids, spermatozoa, lumen and cellular debris. Conversely, no significant differences were found for gross testicular and tubular characteristics, spermatogonia, Sertoli and Leydig cells and intertubular compartments. It was concluded that a single dose of acyline reversibly impaired spermiogenesis, spermatocytogenesis and sperm motility for 2 weeks.