Effect of focused and radial extracorporeal shock wave therapy on equine bone microdamage

OBJECTIVES : To determine whether bone microcracks are altered after application of focused and radial extracorporeal shock wave therapy (ESWT) to the equine distal limb. STUDY DESIGN : An ex vivo experimental model. SAMPLE POPULATION : A contralateral limb specimen was obtained from 11 Thoroughbred racehorses with a unilateral catastrophic injury. Distal limb specimens were also obtained from 5 non-racing horses. METHODS : Three separate skin-covered bone segments were obtained from the mid-diaphysis of the metacarpus (MC3) or metatarsus (MT3). Focused (9,000 shockwaves, 0.15 mJ/mm2, 4 Hz) and radial (9,000 shockwaves, 0.175 mJ/mm2, 4 Hz) ESWT treatments were randomized to the proximal and distal segments and the middle segment was used as a treatment control for pre-existing microcracks. After treatment, bone specimens were bulk-stained with basic fuchsin and microcracks were quantified in transverse calcified bone sections. RESULTS : ESWT had small but significant effects on microcracks. Microcrack density (Cr.Dn) and microcrack surface density (Cr.S.Dn) were increased after focused ESWT, whereas Cr.Le was increased after radial ESWT. In racing Thoroughbreds, Cr.Le increased with increased number of races undertaken. Cr.Dn and Cr.S.Dn were not significantly influenced by the number of races undertaken. CONCLUSION : ESWT has small but significant effects on bone microcracking ex vivo. CLINICAL RELEVANCE : These preliminary data suggest that ESWT has the potential to increase bone microcracking in equine distal limb bone in vivo. Such effects may be more pronounced in Thoroughbreds that are actively being raced, because in vivo microcracking increases with increased number of races undertaken.     

Assessment of Different Functional Parameters of Frozen–Thawed Buffalo Spermatozoa by Using Cytofluorimetric Determinations

Flow cytometry is a useful tool that provides an accurate, objective and rapid evaluation of semen quality. The use of this technique could significantly improve the quality of buffalo semen samples used in artificial insemination. This study was carried out to evaluate, by flow cytometry, frozen–thawed buffalo spermatozoa quality parameters such as sperm viability by SYBR‐14/propidium iodide staining; mitochondrial function by JC‐1 potentiometric probe; sperm chromatin stability (SCSA) by acridine orange; and acrosome reaction (AR) by FITC‐PNA staining. Semen samples from five Italian Mediterranean buffalo bulls were used. Sperm viability was not different between bulls and ranged from 33.4% to 43.6%. A consistent rate (55.1 ± 10.8%) of sperm cells showed high mitochondrial membrane potential (Δψ^high), with no significant differences between subjects. Sperm chromatin structure assay differed significantly between the five buffalo bulls; moreover, data showed high stability within each buffalo. DNA fragmentation indexes (DFI), such as %‐DFI, ‐DFI, SD‐DFI, were 11.2 ± 8.6, 153.3 ± 24.6 and 81.6 ± 21.2, respectively. Regarding AR, the percentage of acrosome‐reacted live (ARL) and acrosome‐reacted dead (ARD) spermatozoa was 0.3 ± 0.2 and 15.3 ± 5.5, respectively. This functional parameter differed significantly between buffalo bulls and showed high stability. Following to Ca²⁺ ionophore A23187 for 3 h, AR significantly differed between subjects and was characterized by an increase in both ARL (10.8%) and ARD population (22.0%). This study indicates that flow cytometry could be a useful tool for a quick multiparametric evaluation of sperm quality in buffalo. In particular, SCSA and AR resulted in sperm functional parameters sensitive enough for the diagnosis of frozen‐thawed semen fertilizing potential.     

Long‐term Effects of Pyrethrin and Cyfluthrin,a Type II Synthetic Pyrethroid,Insecticide Applications on Bull Reproductive Parameters

The objectives of this study were to determine effects of cyfluthrin and pyrethrin spray products, used in combination with cyfluthrin topical and ear tag applications, on bull reproductive parameters over 18 weeks. Angus or Angus x Simmental bulls were randomly assigned to one of three treatment groups: (i) no exposure to pyrethrins/cyfluthrin (CONT; n = 10), (ii) cyfluthrin ear tag and topical applications (ET; n = 10), or (iii) cyfluthrin ear tag, topical, premise spray and pyrethrin fog spray applications (ET+S; n = 8). Bull body weight was measured every 3 week, and body condition score and scrotal circumference were recorded on weeks 0, 9 and 18. Semen and serum were collected every 3 weeks for sperm evaluation and testosterone measurement, respectively. There was a treatment × week interaction (p < 0.01) for sperm with primary defects; bulls in CONT group had a greater (p = 0.01) percentage of sperm with primary defects than bulls treated with insecticides at week 18. Overall and progressive sperm motility, normal sperm morphology, secondary sperm defects and serum testosterone concentrations changed (p < 0.01) over time in all bulls; however, treatment did not affect (p ≥ 0.13) any of these parameters. There were also no treatment effects (p ≥ 0.08) on bull body weight, body condition score or scrotal circumference. The use of pyrethrin‐ and cyfluthrin‐based insecticides, regardless of application, did not negatively affect reproductive parameters in beef bulls when administered over 18 weeks.     

Formation of an Antral Follicle–like Structure of Bovine Cumulus–Oocyte Complexes Embedded Individually or in Groups in Collagen Gels

Culture techniques of antral follicle–like structure (AFLS) derived from cumulus–oocyte complexes (COCs) might provide important insights into follicular development and oocyte maturation. This study was undertaken to investigate the effects of embedding bovine COCs individually (one COC) or in groups (4–5 COCs) in collagen gels on the formation of AFLS and the meiotic status of oocytes. The observations of AFLS formation were performed every second day for 14 days. The AFLS was formed at Day 2 or 4 after the start of culture (Day = 0), irrespective of the culture methods. The mean diameters of AFLS during Days 4–14 using the individual culture method were significantly higher (p < 0.05) than those using the group culture method. However, the AFLS formation rate in the individual culture method was significantly lower compared to that in the group culture method (26.1% vs 62.7%, p < 0.01). Almost all oocytes had undergone the germinal vesicle breakdown stage, irrespective of the culture method or AFLS formation. In conclusion, comparison with the individual culture method revealed that the mean diameters of AFLS in the group culture method were smaller, but more COCs formed AFLS. The group culture method might be useful for evaluating the various hypotheses of follicular formation and interfollicular communication. However, improvement of the group culture system is necessary to prevent the meiotic resumption of oocytes, because the AFLS formation is dependent on the cumulus/granulosa cells surrounding oocytes.     

Fermentation and chemical composition of high‐moisture lucerne leaf and stem silages harvested at different stages of development using a leaf stripper

Lucerne (Medicago sativa L.) requires four or more cuttings at early bud stage per growing season to optimize the amount of crude protein and digestible fibre for feeding high‐producing dairy cows. However, there is potential to generate a nutrient‐dense feed from lucerne regardless of developmental stage by harvesting its protein‐rich leaves separate from its fibrous stems. In order to determine whether fractionated lucerne can be effectively ensiled under high‐moisture conditions and be nutritionally competitive with wilted whole‐plant silage, leaf and stem fractions, harvested at three developmental stages (early bud, 10%–20% bloom and >50% bloom), were directly ensiled in mini‐silos. At day 0, 1, 3, 21 and 140 of ensiling, silages were analysed for protein and non‐protein nitrogen fractions as well as their fermentation products and carbohydrate composition. Silages from unwilted leaves and stems were more heterofermentative than wilted whole‐plant silages; their fermentation shifted from primarily lactic acid to acetic acid production after 21 days. In leaf silages, the high degree of protein degradation into non‐protein nitrogen (~55%) was most likely due to fermentation quality. Nevertheless, at 140 days of ensiling, leaf silages had 21%–25% higher (p < 0.01) available protein (peptide amino acids, soluble and insoluble protein) content than wilted whole‐plant silages, regardless of developmental stage. Achievement of a more rapid pH decline and improved fractionation may further increase the nutritional value of leaf silages.     

菜籽粕可替代含可溶物酒糟饲喂泌乳奶牛

试验旨在确定饲喂含菜籽粕作蛋白质添加物替代全部或部分含可溶物酒糟日粮的反应。试验采用4×4拉丁方设计,12头泌乳荷斯坦奶牛(4头初产、8头经产),试验期4周,在第3和第4周收集数据。配合日粮时,添加菜籽粕100%、66%、33%、0分别替代相应数量的玉米酒糟可溶物(DDGS)。所有日粮含饲草55%(其中玉米青贮50%、苜蓿干草50%)、精料45%,平均含粗蛋白15.1%、乙醚抽提物4.5%。试验结果表明,所有日粮的干物质采食量(25.4kg·d-1)相似。所有日粮的产奶量相似,添加菜籽粕100%、66%、33%、0日粮的产奶量分别为35.2、35.8、34.5、34.3kg·d-1,但菜籽粕含量高的日粮产奶量较高。所有日粮乳蛋白率(3.04%)、乳脂率(3.92%)和乳脂量(1.37kg·d-1)相似,而乳蛋白量则随着日粮中菜籽粕含量的增加而提高,100%、66%、33%、0菜籽粕日粮分别为1.08、1.10、1.05、1.03kg·d-1。所有日粮的饲料转化率相似,每千克干物质采食量含能量校正乳1.46kg。饲喂菜籽粕或DDGS时,赖氨酸是乳蛋白合成的第一限制性氨基酸,而菜籽粕和DDGS同时饲喂时,蛋氨酸为第一限制性氨基酸。所有日粮的瘤胃内容物氨和挥发性脂肪酸浓度相似。试验结果表明,在奶牛日粮中,菜籽粕是DDGS的一种适宜替代物。