Musculoskeletal responses of 2-year-old Thoroughbred horses to early training. 8. Quantitative back-scattered electron scanning electron microscopy and confocal fluorescence microscopy of the epiphysis of the third metacarpal bone

AIM: To characterise and explain the increase in density evident by computerised tomography (CT) and radiography in companion studies as a response to training, in bone in the palmar and dorsal regions of the condyles of the third metacarpal bone (Mc3) of 2-year-old Thoroughbred horses.

METHODS: Compositional back-scattered electron (BSE) imaging in scanning electron microscopy (SEM) and confocal scanning laser microscopy (CSLM) were conducted on polymethyl methacrylate (PMMA)-embedded mediolateral slices of the right distal Mc3 from seven 2-year-old Thoroughbred horses trained on a racetrack and seven untrained horses kept at pasture. One left Mc3 from each group was studied in transverse section planes. This study focussed on regions of Mc3 found to differ in density between the trained and untrained horses in companion studies using CT and radiography.

RESULTS: The increase of bone density in the condyles of Mc3 in trained horses compared with untrained horses occurred, without prior osteoclastic resorption, via the deposition of new bone on pre-existing internal surfaces. Within prior marrow spaces of cancellous bone, there was also rapid formation of immature strands and fronds of bone which were more cellular and mineralised, and more lamellar bone tissue was deposited on these new scaffolding elements in the trained horses. Both resulted in increased bone volume fraction (BVF). The microscopic mineralisation density of the bulk of the new tissue was lower than in pre-existing bone, and CT and radiography underestimated the increase in BVF. The new tissue was thus probably less stiff at the microscopic scale than pre-existing bone, though its addition would stiffen the global structure.

CONCLUSIONS: In Mc3 of all the trained horses, there were obvious differences in microscopic structure compared with those from the untrained horses. Moderate, industry-standard levels of exercise used to prepare young horses for racing induced the formation of new bone in non-bone spaces in bone tissue, such that the bone organ should better withstand later increased levels of exercise.     

Musculoskeletal responses of 2-year-old Thoroughbred horses to early training. 1. Study design,and clinical,nutritional, radiological and histological observations

AIMS: This is the first in a series of papers reporting studies in 2-year-old Thoroughbred racehorses that aimed to determine the response of musculoskeletal tissues to early training on grass and sand racetracks. In this paper, the experimental set-up of the whole study is described, and nutritional, workload, and clinical, radiographic and pathological outcomes are reported, including semi-quantitative assessment of macroscopic changes in articular cartilage.

METHODS: The study group comprised 14 two-year-old Thoroughbred fillies reared entirely at pasture. Of these, seven were selected by a licensed racehorse trainer to undergo a 4-week period of initial training in which they were taught to accept saddle and rider, followed by a 13-week period of flatrace training at a racetrack (Weeks 1–13); the other seven fillies were confined to large grass enclosures and were not trained. Nutrient, including macro- and trace-element intakes were estimated. Distances cantered or galloped and average velocities were quantified for the trained horses. All horses were observed daily, weighed approximately weekly, and underwent a clinical lameness examination at Weeks 5, 9 and 13. Distal forelimbs were radiographed prior to Week 1, during Weeks 7–8, and again at the end of the study, when macroscopic changes in articular cartilage of the proximal surface of the proximal phalanx were also scored after staining with India ink.

RESULTS: Dietary intakes met or exceeded recommended requirements for all nutrients except sodium, which was low in the trained horses. Bodyweight increased throughout the study in the untrained horses, and increased until Week 7 and then decreased slightly in the trained horses. Mean velocity data were used to define three stages of the training programme: Stage 1 comprised canter in Weeks 1–4; Stage 2 comprised canter in Weeks 5–8; and Stage 3 comprised canter in Weeks 9–13 and galloping twice weekly. Four of seven horses completed training. These covered a mean distance of 179.2 km at mean velocities (excluding gallops) of 7.63 m/sec (SD 0.58), 8.99 m/sec (SD 0.56), and 8.43 m/sec (SD 0.74) for Stages 1–3, respectively, and galloped 4.45 km at 14.4 (SD 0.1) m/sec. The three horses that did not complete training became lame in Weeks 9, 10 and 11, and covered 147.9 km at velocities of 7.38 m/sec (SD 0.44), 8.88 m/sec (SD 0.33) and 8.43 m/sec (SD 0.59) and galloped 2.1 km. Overall, slight or intermittent lameness in trained horses was noted on 76/655 (12%) of horse observation days. Swelling was evident on 284/655 (41%) of horse observation days in the metacarpophalangeal (MCPJ) and metatarsophalangeal (MTPJ) joints (92%), palmar metacarpal tendon region (7%) or carpus (1%). Swelling of the MCPJ or MTPJ was not associated with obvious lameness. Radiographic changes were minor and no gross lesions in bone or tendon tissue were evident except for one case of dorsal metacarpal disease. Post mortem, the cartilage of some MCPJ and MTPJ had obvious wear lines and high lesion scores, which were not consistently related to clinical evidence of pain, lameness or joint swelling. Mean lesion scores were not significantly different between the MCPJ and MTPJ, or between trained and untrained horses.

CONCLUSIONS: Workload can be readily quantified in racehorses under semi-commercial training conditions. Obvious lesions in cartilage of the MCPJ or MTPJ were present in some trained and some untrained horses and not consistently associated with clinical evidence of lameness, joint swelling or change in other connective tissues.

CLINICAL RELEVANCE: Workload data in racing horses are likely to be highly relevant for studying the pathogenesis of changes in bone, tendon and cartilage during training, for training management and for risk analysis in racehorse populations. Although obvious cartilage lesions produced little clinical effect, such lesions have previously been shown to be progressive and to prejudice athletic capability. Detection of such occult lesions in young horses will require more sophisticated detection methods.     

高压/酶解/闪式高速提取对金华火腿骨呈味物质的影响

以金华火腿骨为研究对象,研究不同提取方法对其呈味物质释放的影响。通过感官评价及可溶性糖、有机酸、5′-核苷酸和游离氨基酸的测定分析不同处理组间样品的差异,同时采用偏最小二乘回归法（partial least squares regression,PLSR）对提取物感官和呈味物质间进行相关性分析。感官评价发现,高压蒸煮的样品的鲜味、咸味和可接受度最高,其协同滋味的综合评分最佳;呈味物质分析发现,原液中所有呈味物质含量都最低,高压-复配酶解处理的样品中可溶性糖总量最高,高达132.68 mg/100 g;经过酶解处理样品有机酸含量显著增加（P<0.05）,达到3 733.32 mg/100 g;样品经不同处理后,5′-核苷酸含量呈现显著性增加（P<0.05）,总含量最高的是经过高压蒸煮处理的样品,质量分数高达1.24 mg/100 g,是原液和其他处理组样品的2.38～12.4倍;与样品原液相比,游离氨基酸总量都显著增加（P<0.05）,高压蒸煮处理的样品中含量最高,总质量分数为642.44 mg/100 g。借助偏最小二乘回归方法对样品的感官属性及呈味成分进行相关性分析,发现样品间具有显著性差异（P<0.05）,不同的处理方式所释放的呈味物质分布规律显著不同,高压蒸煮的样品与咸味和鲜味具有显著相关性（P<0.05）;酶解处理的样品与酸味和甜味有较强相关性,其他组样品与感官与呈味属性相关性不强。结论：不同处理方式的金华火腿骨具有不同的风味,其风味特征与多种物质有关,该研究结果为火腿骨的深度开发提供了理论指导,提高金华火腿骨的附加值。     

基于酶抑制法的自助式农药残留检测平台建立

以金华火腿骨为研究对象,研究不同提取方法对其呈味物质释放的影响。通过感官评价及可溶性糖、有机酸、5''-核苷酸和游离氨基酸的测定分析不同处理组间样品的差异,同时采用偏最小二乘回归法（partial least squares regression,PLSR）对提取物感官和呈味物质间进行相关性分析。感官评价发现,高压蒸煮的样品的鲜味、咸味和可接受度最高,其协同滋味的综合评分最佳;呈味物质分析发现,原液中所有呈味物质含量都最低,高压-复配酶解处理的样品中可溶性糖总量最高,高达132.68 mg/100 g;经过酶解处理样品有机酸含量显著增加（P<0.05）,达到3 733.32 mg/100 g;样品经不同处理后,5''-核苷酸含量呈现显著性增加（P<0.05）,总含量最高的是经过高压蒸煮处理的样品,质量分数高达1.24 mg/100 g,是原液和其他处理组样品的2.38～12.4倍;与样品原液相比,游离氨基酸总量都显著增加（P<0.05）,高压蒸煮处理的样品中含量最高,总质量分数为642.44 mg/100 g。借助偏最小二乘回归方法对样品的感官属性及呈味成分进行相关性分析,发现样品间具有显著性差异（P<0.05）,不同的处理方式所释放的呈味物质分布规律显著不同,高压蒸煮的样品与咸味和鲜味具有显著相关性（P<0.05）;酶解处理的样品与酸味和甜味有较强相关性,其他组样品与感官与呈味属性相关性不强。结论：不同处理方式的金华火腿骨具有不同的风味,其风味特征与多种物质有关,该研究结果为火腿骨的深度开发提供了理论指导,提高金华火腿骨的附加值。     

Charcoal and smoke extract stimulate the soil microbial community in a highly weathered xanthic Ferralsol

The influence of charcoal and smoke condensates (pyroligneous acid, PA) on microbial activity in a highly weathered Amazonian upland soil was assessed via measurements of basal respiration (BR), substrate-induced respiration (SIR), and exponential population increase after substrate addition. PA extracts are commonly used for fertilizer or as pest control in Brazil, where phosphorus (P) availability and nitrogen (N) leaching are among the most severe limitations for agriculture. Microbes play an important role in nutrient cycling and solubilizing of phosphate. BR, microbial biomass, population growth and the microbe's efficiency (expressed by the metabolic quotient) increased linearly and significantly with increasing charcoal concentrations (50, 100 and 150 g kg⁻¹ soil). Application of PA caused a sharp increase in all parameters. We suppose that the condensates from smoke contain easily degradable substances and only small amounts of inhibitory agents, which could be utilized by the microbes for their metabolism.     

饲料中肉骨粉含量的近红外反射光谱检测方法

收集28份鸡饲料,31份猪饲料,25份牛饲料和肉骨粉7份,在饲料中掺入不同比例(0.5%～6.0%)的肉骨粉,制成分析样本.采用偏最小二乘(PLS)法,分别对光谱进行散射校正、平滑、一阶导数和二阶导数预处理,建立了鸡饲料、猪饲料和牛饲料中肉骨粉含量的近红外定量分析模型.利用验证集样本对定标模型进行了检验,鸡饲料、猪饲料和牛饲料中肉骨粉含量的真值与预测值之间的决定系数分别为0.9694、0.9846和0.9788;标准差分别为0.279、0.252和0.287;相对分析误差分别为5.663、6.865和5.889.结果表明,利用近红外光谱法测定饲料中的肉骨粉含量具有较高的预测精度.     

Surgical repair of radial carpal bone fractures in two racehorses

A 3-year-old Standardbred gelding (Case 1) and a 2-year-old Thoroughbred gelding (Case 2) were referred for surgical evaluation of a left radial carpal bone (RCB) fracture, sustained during training. Clinical findings at the time of initial examination included a palpable effusion within the left middle carpal joint in both horses and marked signs of pain and reduced range of motion on flexion of the affected carpus. In both horses, the RCB fracture was evident on the following radiographic views of the carpus: dorsolateral–palmaromedial oblique (30° off lateromedial) and flexed lateromedial. An additional loose wedge-shaped osteochondral fragment at the proximal articular surface of the RCB could be seen in Case 2. Both horses underwent surgical reduction and repair of the fracture between 1 and 2 days following the initial injury, which consisted of arthroscopic removal of any intra-articular osteochondral fragments, and arthroscopic assisted-interfragmentary compression via a standard dorsomedial and dorsolateral approach to the antebrachiocarpal joint (ACJ) and middle carpal joints (MCJ). The two horses returned to function as racehorses, 6 months (Case 1) and 16 months (Case 2) after surgery. The RCB is a relatively uncommon site for large carpal fractures in horses. The clinical presentation and findings from this report were similar to that of third carpal bone (C3) slab fractures, confirming that surgical repair is indicated in selected cases of RCB fractures.     

Congenital Osteoma of the Frontal Bone in an Arabian Filly

Congenital frontal osteoma has not been previously described in horses. This report records—for the first time—a congenital osteoma of the frontal bone in a 4-month-old Arabian filly. The filly had a frontal hard mass that was present at birth and then showed a slow and continuous growth. This mass appeared as a solitary, painless, oval dense tumor of compact bone, about 2 cm in diameter and 3 cm in length. The tumor was asymptomatic, and the skin over the mass was normal. Radiography revealed a well-defined oval, radio-dense mass projecting from the surface of the right frontal bone with no local invasion. The tumor had a broad-based attachment to frontal bone with normal frontal sinus. The mass caused disfigurement; therefore, it was removed at the owner’s request. The mass was diagnosed histopathologically as osteoma. The surgical excision of the osteoma was successful without any complications, and the filly adapted remarkably well after surgery. No recurrence was reported 20 months after the surgery. In conclusion, osteoma should be listed during the differential diagnosis of the congenital craniofacial masses in horses. Early diagnosis of the frontal osteoma guarantees a successful surgical treatment and consequently prevents the future complications.     

Relationships between body size and secondary sexual characters,and sperm characters in male Dolly Varden char (Salvelinus malma)

In Salmonidae, subordinate males are exposed to higher risks of sperm competition than dominant males and thus are expected to improve the sperm characteristics (sperm concentrations, sperm velocity and sperm longevity). In this study, we investigated the relationships between body size and secondary sexual characters (breeding colour, hump height and snout length), and sperm characteristics of one‐year‐old (newly matured) Dolly Varden char. Small males displayed higher sperm concentrations than large males. Moreover, males with dull breeding colours, but not with lesser snout length and hump height, displayed an increased sperm velocity compared to males with bright colours, suggesting a trade‐off between sperm quantity and the investment in breeding colour. In addition, sperm longevity decreased as sperm swimming velocity increased. These findings indicate that small males with dull breeding colours improve the quantity and quality of their sperm to a great extent to enhance their chances of reproductive success.