Osteopetrosis in a neonatal donkey

Osteopetrosis is a rare disorder characterised by a defect in osteoclastic bone resorption. This report describes osteopetrosis in a neonatal donkey that suffered a displaced tibial fracture. Radiographic examination identified generalised reduction in medullary cavity size, thickened mid‐diaphyseal cortices and conical metaphyseal bone extending toward the mid‐diaphysis of long bones. Postmortem examination identified additional fractures and brittle bones. Histologically, osteoclasts were absent in multiple bone sections. Diaphyseal cortices consisted of concentric bone lamellae with marrow tissue infiltration. Large wedges of secondary spongiosa extended from the metaphyseal growth plate. Clinical and histopathological features were similar to an osteoclast‐deficient, autosomal recessive form of osteopetrosis in humans.     

施用活性硅降低重金属移动性的研究

该研究通过一系列的土柱实验研究土壤中硅和重金属的反应机制。土柱实验在可溶性镉、铜、镍和铅酸盐的情况下,用各种活性硅（硅藻土,沸石,无定形二氧化硅,浓缩单硅酸）处理灰森林土。所有富硅物质都用电子显微镜测试分析过。富硅物质对土壤中重金属的钝化试验结果表明：硅藻土和浓缩单硅酸比沸石和无定形二氧化硅更能降低重金属的移动性。重金属移动性的降低是由土壤溶液中单硅酸和重金属的反应,以及富硅物质表面对重金属的吸附实现的。重金属在土壤中移动的强度与自身种类有关。施用富硅物质可明显降低镉和镍的移动性,对铜和铅的效果不明显。     

Motility Characteristics and Fertilizing Capacity of Boar Spermatozoa Stained with Hoechst 33342

Flow cytometry sorting of spermatozoa using fluorescence dye Hoechst 33342 is the only effective sex selection methodology validated in numerous laboratories. This study was carried out to determine the effect of Hoechst 33342 on the motility and fertility of stained boar spermatozoa. Experiment 1 evaluated motility parameters (percentage of motile spermatozoa, velocity, angularity and oscillation) of boar spermatozoa stained with Hoechst 33342 by a computer‐aided sperm analysis (CASA) instrument. Spermatozoa (30 million/ml) were divided into five treatment groups and stained during 1 h at 35°C with 9, 18, 27, 60 and 90 μM of H33342. There were no differences in sperm motility patterns nor percentages of motile spermatozoa incubated in the presence of 9, 18 or 27 μM. Percentage of motile spermatozoa and motility parameters decreased significantly (p < 0.05) at 60 μM of Hoechst 33342. Spermatozoa were immotile at concentration of 90 μM. In experiment 2, pregnancy rates, farrowing rates and litter size from sows (n = 275) artificially inseminated (AI) with either Hoechst 33342 stained (27 μM) or unstained (control) spermatozoa were determined. Sows inseminated with stained spermatozoa had no significant lower pregnancy rate (88.33%) as compared with controls (90.32%). Staining neither affected farrowing rates (85.0 vs 87.7%) nor total number of piglets born (10.56 ± 0.32 vs 10.47 ± 0.24, stained and controls, respectively). No phenotypical abnormalities were registered among the newborn piglets. The data suggest that incubating spermatozoa with Hoechst 33342 at levels required for X‐ and Y‐bearing chromosome sperm sorting, does not impair sperm viability or their fertility after AI.     

In vitro Maturation of Oocytes from Santa Ines Ewes Subjected to Consecutive Sessions of Follicular Aspiration by Laparoscopy

The success of embryo production in vitro depends upon the use of an efficient oocyte retrieval technique, and the best results have been obtained by laparoscopic aspiration. The aim of this study was to evaluate the effect of consecutive sessions of follicular aspiration on the quantity, quality and in vitro maturation competence of oocytes obtained from ewes subjected to hormonal stimulation. Six Santa Ines ewes underwent nine sessions of follicular aspiration by laparoscopy with a 7‐day interval between sessions, totalling 56 aspirations. After 24 h of culture, oocytes were stained and classified according to the stage of nuclear and cytoplasmic maturation. Oocyte retrieval rate was 61.4 ± 2%, resulting in a total of 249 oocytes. No significant variation was observed between sessions (p > 0.05). The average number of oocytes retrieved from each ewe was 6.4 ± 2 per session and 42 ± 4 in total. No significant difference was observed between the frequencies of the different stages of nuclear maturation: 32.72% mature, 40.74% immature and 26.54% degenerated/indeterminate oocytes; however, a significant difference was observed between the frequencies of the different stages of cytoplasmic maturation: 10.7% mature, 73.25% immature and 16.05% degenerated/indeterminate oocytes. No significant difference was observed in nuclear or cytoplasmic maturation between the weeks of procedure. We conclude that after nine consecutive sessions of follicular aspiration, the quantity and quality of retrieved oocytes remained unchanged as well as the levels of nuclear and cytoplasmic maturation obtained, demonstrating the viability of this technique for repetitive follicular aspirations on the same donor.     

Endometrial Explant Culture to Study the Response of Equine Endometrium to Insemination

Mating‐induced endometritis (MIE) is ubiquitous in the horse after natural mating and artificial insemination with frozen/thawed semen causing the most aggressive response. The majority of mares eliminate MIE 24–48 h after insemination. An endometrial explant culture was tested as a potential in vitro exemplar for sperm‐induced MIE. Endometrial prostaglandin F_2α (PGF_2α) secretion and expression of interleukin‐8 (IL‐8) were used as markers of inflammation. Endometrial explants were cultured from uteri collected from follicular phase mares. Explants were challenged with 1 or 10 × 10⁶ sperm/ml frozen/thawed semen, chilled semen, washed sperm or seminal plasma. Medium was collected 24 and 72 h after challenge and assayed for PGF_2α by radioimmunoassay. Treatment of endometrial explants with frozen/thawed, chilled semen or washed sperm did not change the secretion of PGF_2α compared with untreated controls. However, 24 h after challenge cultured explants expressed IL‐8. The in vitro endometrial explant system did not represent the in vivo response to semen when PGF_2α was used as a marker of inflammation, yet the use of gene expression as an inflammatory marker warrants further investigation.     

The Effect of Two Antibiotic Mixtures on Haemophilus somnus, Campylobacter fetus ssp. venerealis, Mycoplasma bovis, and Ureaplasma diversum in Frozen Bovine Semen

A two-step semen-extending protocol was compared to a one-step protocol in its efficiency in inhibiting growth of Haemophilus somnus, Campylobacter fetus ssp. venerealis, Mycoplasma bovis , and Ureaplasma diversum in experimentally infected semen. In both protocols, the effect of an antibiotic mixture of 500 μg gentamycin, 100 μg tylosin, 300 μg lincomycin, and 600 μg spectinomycin (GTLS) was compared to a mixture of 500 IU penicillin, 500 IU streptomycin, 150 μg lincomycin, and 300 μg spectinomycin (PSLS). The one-step extending method was as effective as the two-step extending method. Both antibiotic mixtures were equally effective in controlling C. fetus . For H. somnus and U. diversum , the PSLS mixture was more effective than the GTLS mixture. It was striking that both antibiotic mixtures had no effect in decreasing the numbers of M. bovis .     

Effect of Oxytocin Treatment on Artificial Insemination with Frozen–Thawed Semen in Murciano–Granadina Goats

The site where the semen is deposited appears to be one of the most important factors affecting pregnancy of inseminated goats. In Murciano–Granadina (MG) goats, post-cervical insemination is achieved in a limited number of females. An effective way to increase fertility rate could be by increasing post-cervical inseminations. Effect of exogenous oxytocin application to facilitate the cervical penetration and its effect on kidding rate and prolificacy in MG goats were investigated. Oestrus was synchronized using progesterone-impregnated sponges for 11 days. Females were randomly divided into three groups (n = 190) and received either an i.v. injection of 100 or 200 IU of oxytocin or saline solution 15 min before being inseminated. Data on semen deposition depth were recorded for each animal using a catheter scaled in centimetres (up to 4 cm). Depth of semen deposition was affected by the oxytocin treatment (p < 0.05). Oxytocin enhanced cervical passage only with the dose of 200 IU compared with the control group, increasing the deposition depth (2.9 cm vs 1.9 cm). No significant effect of oxytocin treatment on kidding rate and prolificacy was detected. Depth of semen deposition affected kidding rate (p < 0.01). In conclusion, oxytocin treatment improved the depth of semen deposition in AI of MG goats, but kidding rate and prolificacy was not affected. More studies must be conducted to assess the minimal effective dose required for sufficient cervical dilation, and to determine the effects of such doses of oxytocin on uterine motility, sperm transport and fertility in goats.     

The effect of medetomidine and its antagonism with atipamezole on stress-related hormones, metabolites, physiologic responses, sedation, and analgesia in goats

Six 3‐year‐old goats (three males and three females) weighing 60.0 ± 18 kg (mean ± SD) were used to investigate the effect of medetomidine (MED; 20 µg kg^?1 IV) and its antagonism with atipamezole (ATI; 100 µg kg^?1 IV) on physiologic responses (heart rate (HR; beats minute^?1), respiratory rate (RR; breaths minute^?1), electrocardiogram (ECG), rectal temperature (T; °C), blood pressure (oscillometric; mm Hg), sedation (SED), posture (REC), analgesia (ALG), and stress‐related hormonal and metabolic responses (epinephrine and norepinephrine (high performance liquid chromatography with electrochemical detection), cortisol (COR; µg dL^?1; radioimmunoassay), glucose (GLU; mg mL^?1; enzymatic colorimetric assay), and free fatty acids (modified enzymatic colorimetric assay)); each goat received ATI or SAL in random order separated by 1 week. Jugular catheters were placed for drug administration and blood sampling (10–12 mL sample^?1) using a lidocaine skin block (20 mg) 2 hours prior to beginning of each trial; during this trial, goats breathed room air. Physiologic parameters were measured, SED, REC, and ALG were scored, and blood samples were collected from jugular catheters at baseline (time = ?30 minutes), 5 minutes post‐MED administration (time = ?25 minutes), 25 minute post‐MED administration and immediately prior to antagonism (time = 0 minute), and at 5, 30, 60, and 120 minutes after administering ATI or SAL. ALG was tested by clamping the withers and metacarpus with hoof testers fitted with a force transducer to measure applied isometric force (lb) (a technique used previously in goats to evaluate analgesia). Continuous variables were analyzed by Repeated Measures analysis of variance (anova ); categorical data were analyzed using a Friedman Repeated Measures anova on ranks. A p‐value of <0.05 was considered significant. If a significant difference was found, a Dunnett's pair‐wise comparison of means was conducted. Differences between ATI and SAL were examined at 5, 30, 60, and 120 minutes using a paired t‐test with a Bonferroni correction. Administration of MED resulted in a decrease in T (38.7 ± 0.3 to 34.5 ± 0.4 °C), HR (78 ± 19 to 55 ± 9), and RR (31 ± 12 to 14 ± 5) over time; an increase in mean arterial blood pressure (90 ± 19 to 132 ± 23), COR (0.254 ± 0.125 to 4.327 ± 1.233), and GLU (82.0 ± 13.2 to 255.9 ± 38.9); and changes in SED (alert to marked sedation), REC (standing to recumbent), and ALG (metacarpus = 5 ± 2 to 14 ± 0; withers = 3 ± 2 to 14 ± 0). GLU was 62–70% higher at 60 and 120 minutes and COR was 336% higher after SAL than after ATI at 120 minutes; at 30, 60, and 120 minutes, T was 4–10% higher after ATI than SAL. There were no other significant differences. REC, SED, and ALG were antagonized after ATI. ATI did not antagonize the effect of MED on HR, RR, or MAP, but stabilized T and antagonized the increase in GLU and COR.