Quantitative analysis of the optic nerve of the horse (Thoroughbred)

Three optic nerves (L1, R2, R3) 12-18 mm behind the eyeball of the horse (Thoroughbred) were investigated quantitatively under light and electron microscopes. Thin sections at the thickness of 0.35 microm were cut, stained by toluidine blue and observed under the light microscope. The areas of the optic nerve and the axon bundles were 20.03 +/- 1.04 and 16.59 +/- 0.79 mm2 (mean +/- SD, n=3), respectively. The axon numbers for optic nerve L1, R2 and R3, estimated from light micrographs, were about 481 x 10(3), 543 x 10(3), and 494 x 10(3), respectively. Axons of optic nerve L1 were also counted from electron micrographs and the total number of 488 x 10(3) was received. Furthemore, axon diameters of optic nerve L1 were also measured from electron micrographs. The diameter of a circle with the same peripheral length as an axon, was regarded as its diameter. The medullary sheath of the axon was not included during measuring. Altogether 5,744 axons were measured and axon diameters were in a range of 0.23-12.69 microm, with a mean of 2.56 +/- 1.45 microm (mean +/- SD). A regional difference of axonal diameters was found across the optic nerve: the mean diameter of axons in the centrodorsal region (2.28 microm) was the smallest, and had significant difference with those in several peripheral regions (P<0.05).     

Quantitative motor unit action potential analysis of skeletal muscles in the Warmblood horse

Motor unit action potential (MUP) analysis in human medicine is a valuable and important diagnostic technique enabling discrimination between myogenic and neurogenic problems. This study establishes normative data in subclavian, triceps and lateral vastus muscles for clinical application of MUP analysis in the Warmblood horse, and examines whether muscle differences are present. Electromyographic (EMG) needle examination and MUP analysis were performed of the triceps, lateral vastus and subclavian muscles in 7 awake, nonsedated, Warmblood horses age 4-10 years. The amplitude, duration, number of phases and turns were calculated from the recorded superimposed MUPs together with intramuscular and rectal temperatures. No significant differences were found in duration of insertional activity between the 3 muscles. The mean +/- s.d. duration of the insertional activity was 526 +/- 1483 ms. The MUP amplitude of all 3 muscles differed significantly, with the highest amplitude (427 +/- 3.20 microV) in the triceps and the lowest (220 +/- 2.08 microV) in the subclavian muscle. The number of turns of the lateral vastus (3.0 +/- 1.22) was significantly higher than that of the triceps muscle (2.7 +/- 1.51). No differences were found in MUP duration (5.9-6.4 ms).     

Quantitative analysis of the organization of the inner nuclear layer of the horse retina

Four cell types including the bipolar, amacrine, horizontal and Muller cells were investigated quantitatively in the inner nuclear layer of the retina in the horse. Cells were identified on the basis of the morphology and distribution of processes leaving from their somata, cytological features and positional features. The average percentages of the above 4 cell types were 44%, 24%, 1% and 29%, respectively. The average total cell densities in the inner nuclear layer in the visual streak, the nasal and temporal regions, the dorsal and ventral regions of the retina were also estimated. It is expected that the results of this study will supply the basic data for further study of the neural circuits in the horse retina.     

广西百色马生长激素基因的克隆与序列分析

为了丰富优良地方品种百色马的分子遗传学基础数据,试验设计1对特异性引物,利用PCR技术成功克隆百色马的生长激素基因全序列.结果表明:百色马GH基因碱基序列全长1 922 bp(GenBank登录号为FJ604598),包含完整的5个外显子和4个内含子;与蒙古马比较,百色马GH基因DNA序列出现12个碱基的变异,有3个变异位点处于编码区,均为有义突变,突变方式都为G→A的转换,其中第981位碱基G→A致使第89位氨基酸由Arg突变为Lys,第1739位碱基G→A使第185位氨基酸由Gly→Arg,第1 764位碱基G→A使第193位氨基酸由Gly→Asp.基于GH基因构建的不同,动物亲缘进化树的拓扑结构与物种树一致,哺乳动物的GH基因在进化中比较保守.     

Histopathologic study of long-bone growth plates confirms the basset hound as an osteochondrodysplastic breed

Osteochondrodysplasias are caused by abnormal development and growth of cartilage and bone. These abnormalities have been reported in both humans and animals with dwarfism. The basset hound is considered a breed with a disproportionate prevalence of dwarfism, the cause of which is unknown. To determine the type of osteochondrodysplasia in this breed, we analyzed histologically the growth plates from the long bones of a basset hound and a Doberman pinscher, both 2 mo old. Tissue was fixed in 4% paraformaldehyde, embedded in paraffin, sectioned at 5 microm, stained with hematoxylin and eosin, and analyzed by light microscopy. Our results suggest that by this method the basset hound can be defined only as a breed having osteochondrodysplasia due to a primary cartilage problem in the growth plate.     

Healed long-bone fractures in wild birds

AIM: To build a profi le of the oral lesions that occur in sheep in New Zealand that need to be considered within the differential diagnosis of foot-and-mouth disease.

METHODS: Lesions of the anterior lips and gums of sheep were surveyed in two abattoirs, photographed, and described grossly and histopathologically.

RESULTS: A sequence of lesions in order of age and stage of healing are described and illustrated, and their pathogenesis discussed. Lesions of the midline of the lips and gums of traumatic or irritant aetiology were common, and the prevalence was higher in adult sheep than in lambs.

CONCLUSIONS: The majority of lesions observed appeared to be primarily of traumatic aetiology. They probably arose from the fright/flight response behaviour of sheep, resulting in the mouth impacting against wire fences or yard railings while being handled. A smaller percentage of lesions may have been due to abrasive or irritant feed or soil. The presence of plant material and bacteria in lesions delayed healing and contributed to the formation of ulcers.     

Genetic characterisation of the Uruguayan Creole horse and analysis of relationships among horse breeds

The genetic variability within the Uruguayan Creole horse and its relationship to a group of geographically or historically related breeds (Spanish Pure-bred, Barb, Quarter horse, Paso Fino, Peruvian Paso, Arabian and Thoroughbred horse), was evaluated using 25 loci (seven of blood groups, nine of protein polymorphisms and nine microsatellites) analyzed on a total of 145 Uruguayan Creole horses. In this study, blood group and protein polymorphism variants that are considered to be breed markers of Spanish Pure-bred and Barb horses were detected in the Creole breed. Conversely, some microsatellites and protein polymorphisms alleles were found uniquely in the Creole horse. American horse breeds together with Barb and Arabian horses clearly formed a separate cluster from the Spanish pure-bred and Thoroughbred breeds, as shown by an UPGMA dendrogram based on Nei's standard genetic distance. Data in this study provided evidence for considerable genetic variation within Uruguayan Creole horses and of a distinctive breed profile. Both traits were most likely inherited from the XVIth century Spanish horses, more closely related to Barb than to Spanish Pure-bred.     

Effects of toe angle on hoof growth and contraction in the horse

The effects of toe angle on the growth of the unshod hooves of mature horses were measured over 126 days. The hooves of 4 horses were trimmed long in the toe and short in the heel (“LT”), with toe angles of 40° for the forelimb and 50° for the hind limb hooves; 4 others were trimmed short in the toe and long in the heel (“ST”), with toe angles of 50° for the forelimb and 55° for the hind limb hooves. Growth of the hoof wall at the toe ranged between .19 and .28 mm/day, and was slowest in the forelimb hooves trimmed ST. After 126 days, the hooves trimmed LT were 7% smaller in width than they had been at day 0. Narrowing of the hoof walls and frogs was accompanied by deformation of the angles of the walls (bending outward of their weightbearing surfaces). Frog lengths and sole areas were not affected by toe angle. Regardless of trimming method, all forelimb hooves tended to return to a toe angle of 45° between trimmings, while all hind limb hooves tended toward toe angles of 52° to 53°. The soles of all hooves were basically circular in shape, although the hooves trimmed LT tended to be skewed to the left, as viewed from above, after 126 days.     

A morphometric study of foetal and newborn cardiac growth in the horse

A morphometric study of hearts in 81 equine foetuses, ranging in age from 190 to 330 days of foetal age, and in 26 newborn foals is reported. The mean weight, external dimensions, ventricular wall thickness and circumference of the atrio-ventricular orifice were measured. Features of the main associated vessels of the heart were also recorded. All cardiac measurements of foetuses increased linearly throughout the latter half of pregnancy and were highly correlated with foetal age. This linear growth pattern was also found in the parameters of the associated arterial trunks. The ratio of the right ventricular weight to the total ventricular weight in the equine foetuses increased gradually from 0.27 (Day 190) to 0.34 (full term), but in newborn foals, this ratio began to decrease soon after birth, declining from 0.32 to 0.28 by 11 days of age. The interventricular septum was thickest followed by the right ventricular wall, and the left ventricular wall was thinnest during the period of gestation studied. The right ventricle was 3.2 per cent to 31.5 per cent thicker than the left ventricle.     

Quantitative detection of atropine-delayed gastric emptying in the horse by the 13C-octanoic acid breath test

The 13C-octanoic acid breath test has been correlated significantly to radioscintigraphy for measurement of gastric emptying indices in healthy horses. The objective of this study was to investigate the validity of the test for measurement of equine delayed gastric emptying, prior to its potential clinical application for this purpose. A model of atropine-induced gastroparesis was used. Gastric emptying rate was measured twice in 8 horses using concurrent radioscintigraphy and/or breath test after treatment i.v. with either atropine (0.035 mg/kg bwt) or saline in randomised order. Analysis of both data sets demonstrated that the atropine treatment had caused a significant delay in gastric emptying rate. Paired breath test data showed an atropine-induced delay in gastric half-emptying time (t 1/2), with no overlap in the 99% CI range (P < 0.001). Significant correlations were found between scintigraphy and 13C-octanoic acid breath test for calculation of both t 1/2 (P < 0.01) and lag phase duration (P < 0.05) in the atropine-delayed emptying results. The mean (s.d.) bias in breath test t 1/2 when compared with scintigraphy was 1.78 (0.58) h. The results demonstrated that the 13C-octanoic acid breath test was an effective diagnostic modality for the measurement of equine delayed gastric emptying. The technique offers advantages to existing methods for clinical investigation, as it is noninvasive, not radioactive, quantitative and requires minimal equipment or training to perform.     

Preliminary study of histological comparison on the growth patterns of long-bone cortex in young calf, pig, and sheep

Some young large farm animals show a laminar bone formation in the long-bone cortex. Such a laminar bone is gradually replaced by Haversian bone with osteons during their growth periods. In this preliminary study, we observed the transverse ground samples of tibia cortex in young calves, pigs, and sheep by backscattered electron imaging. The cortex bones of all the newborn (NB) animals were basically formed with laminar bone structures. The NB and 1-month-old (1-M) calves had a typical concentric structure of laminar bone, whereas the NB and 1-M pigs showed a wire-netting bone with laminar-bone units. The NB sheep was similar to the calf rather than the pig. In the growth rate of bone volume, sheep was similar to calf up to 6 months after birth (6-M). Such calf and sheep showed a more rapid ratio of bone volume than pig. A few osteons had initially appeared in the innermost layer of the 6-M calf. A 1-year-old (1-Y) calf showed scattered osteons in the bone cortex, but many laminar-bone units were still retained in the outer layer. A 6-M pig had many osteons in the entire cortex but only a few osteons in the outermost layer. In the 6-M sheep, no osteons were observed, whereas a 1-Y sheep showed a relatively small number of osteons mainly in the middle layer but a higher osteon-volume than the 1-Y calf. In the 1-Y sheep, the more widely absorbed areas by bone-remodeling with osteons were observed as compared with the 1-Y calf, and the bone volume was decreased from the 6-M into the 1-Y sheep because of the remarkable bone-absorption. Thus, calf kept on possessing many laminar-bone units for a longer time in the growth period than sheep, while pig showed the earliest bone-remodeling with osteons. These results may be caused by their different body size and withers height in calf and sheep after growing and the difference of the dependence upon mother's body during juvenile period between pig and calf with sheep. The initial region of osteon formation may be distinguishable among their animals, respectively. However, further detailed investigations of their young animals at successive stages will be necessary.     

Models in vivo of wound healing in the horse and the role of growth factors

Abstract Wound models attempt to simulate the natural healing processes in wounds. However, all models have significant limitations due to the complexity of the tissue repair process. Much can be learned from wound models in vitro by the use of cell culture techniques. The horse can provide a suitable naturally occurring model of chronic wound healing because it has many similarities to wound healing encountered in human medicine. The tissue architecture was investigated with regard to extracellular matrix and growth factor distribution during wound healing and growth factors were consistently present in the wound area. Biochemical investigations revealed increased levels of hydroxyproline, collagen, and TGFβ1 in exuberant granulation tissue. Equine wound models were established in vitro using cell culture techniques and growth factors had significant effects on the growth of the cells and their ability to synthesize collagen. Two gelatinases (MMP-2 and MMP-9) were detected in the tissues and wound fluid samples investigated. Zusammenfassung Wundmodelle haben zum Ziel, die natürlichen Heilungsprozesse in Wunden zu simulieren. Allen Modellen sind durch den komplexen Gewebsheilungsprozess deutliche Grenzen gesetzt. Durch die Verwendung von Zellkulturtechniken kann von Wundmodellen in vitro viel abgeleitet werden. Das Pferd stellt wegen der vielen Ähnlichkeiten zur humanmedizinischen Wundheilung ein geeignetes Modell für chronische Wundheilung dar. Die Gewebsarchitektur wurde bezüglich der Verteilung von extrazellulärer Matrix und von Wachtumsfaktoren während der Wundheilung untersucht; Wachtsumsfaktoren waren ständig in der Wunde vorhanden. Biochemische Untersuchungen ergaben erhöhte Hydroxyprolin-, Kollagenund TGFβ1-Spiegel. Wundmodelle beim Pferd in vitro und Zellkulturtechniken wurden entwickelt; Wachstumsfaktoren hatten deutliche Wirkung auf die Zellen und ihre Fahigkeit zur Kollagensynthese. Zwei Gelatinasen (MMP-2 und MMP-9) wurden im Gewebe identifiziert und Wundflüssigkeitsproben untersucht. [Cochrane, C.A. Models in vivo of wound healing in the horse and the role of growth factors. (Wundheilungsmodelle in vivo beim Pferd und die Rolle von Wachstunisfaktoren). Veterinary Dermatology 1997; 8 : 259–272] Resumen Los modelos de heridas intentan estimular el proceso natural de curación de heridas. Sin embargo, todos 10s modelos presentan limitaciones considerables debido a la complejidad del proceso de curación de heridas. Se puede aprender mucho de modelos de heridas in vitro mediante el uso de técnicas de cultivo celular. El caballo puede suponer un modelo natural adecuado de curación crónica de heridas ya que se asemeja a la curación de heridas en medicina humana. Se investigó la arquitectura tisular en referencia a la matriz extracelular y la distribución de factores de crecimento viendo que los factores de crecimiento se encontraban presentes de forma constante en el área de la herida. Las investigaciones bioquímicas revelaron incremento en los niveles de hidroxiprolina, colágeno y TGFbl en tejido de granulación exhuberante. Se establecieron modelos in vitro de heridas equinas utilizando técnicas de cultivo celular y los factores de crecimiento tuvieron efectos significativos en el crecimiento de las células y su capacidad de sintetizar colágeno. Se detectaron dos gelatinasas (MMP-2 y MMP-9) en 10s tejidos y muestras de fluidos de las heridas investigadas. [Cochrane, C.A. Models in vivo of wound healing in the horse and the role of growth factors. (Modelos de curacion de heridas in vivo en el caballo y el papel de 10s factores de crecimiento). Veterinary Dermatology 1997; 8 : 259–272] Résumé Les modèles de plaies tentent de simuler les processus naturels de cicatrisation. Cependant tous les modèles ont des limitations significatives dues à la complexité des processus de réparation des tissus. Beaucoup peut être étudié sur des modèles de plaies in vitro suite à l'utilisation des techniques de culture cellulaire. Le cheval peut constituer un modèle nature1 fiable de cicatrisation chronique de plaie de par les nombreuses similarités qu'il partage avec la cicatrisation des plaies en médecine humaine. L'architecture tissulaire a étéétudiée concernant la matrice extracellulaire et la distribution des facteurs de croissance pendant la cicatrisation, et les facteurs de croissance se sont montrés présents de façon consistante dans la zone de la plaie. Des investigations biochimiques ont révélé une élévation des taux d'hydroxyproline, collagène, et TGFβ1 dans la granulation tissulaire exubérante. Des modèles équins de plaies ont étéétablis in vivo en utilisant des techniques de culture cellulaire et les facteurs de croissance ont montré des effets significatifs sur la croissance des cellules et leur capacité de synthétiser le collagène. Deux gélatinases (MMP-2 et MMP-9) ont été détectées dans les tissus et les échantillons de fluide des plaies ont été analysés. [Cochrane, C.A. Models in vivo of wound healing in the horse and the role of growth factors. (Modeles in vivo de cicatrisation des plaies chez le cheval et role des facteurs de croissance). Veterinary Dermatology 1997; 8 : 259–272]     

Rectal tears in the horse: an analysis of 35 cases

The records of 35 horses with Grade 3 or 4 rectal tears, presented to the Veterinary Medical Center at Texas A & M University over a five year period, were reviewed. Grade 3 tears were sub-classified according to whether the remaining tissue was serosa (Grade 3a) or mesorectum (Grade 3b). Five horses were destroyed on presentation and 30 were treated by primary suture closure (8 horses), faecal diversion alone (9 horses) or in combination with suture closure (11 horses) and packing of the tear with medicated gauze sponges (two horses). Faecal diversion was achieved with a temporary indwelling rectal liner (TIRL) in 19 horses and colostomy in one. Survival was related to classification of the tear, efficacy of first aid measures administered at time of injury and method of treatment. Seventy-four per cent of horses with Grade 3a tears and 44 per cent of those with Grade 3b tears survived. Grade 4 tears had a grave prognosis. Horses given adequate first aid before admission had a better survival rate. With proper patient selection, primary closure of the tear with sutures yielded excellent results. In horses which were not candidates for suture closure alone, a combination of faecal diversion and suturing gave better results than faecal diversion only. In addition, selected horses were treated successfully by packing the rectal tear with gauze sponges. The results demonstrate the value of a TIRL to divert faeces and appropriate first aid measures in treating rectal tears.     

Bioelectrical impedance analysis determination of water content and distribution in the horse

A horse’s hydration status is critical to its health. The accurate and quantitative determination of it has been problematic because of size, length and density of hair, and uneven topography. The objective of this study was to validate a bioelectrical impedance analysis (BIA) method for objectively quantifying hydration status. Monofrequency BIA values and simple biometric measurements were used to construct predictive equations for total body water, plasma, extracellular, and intra-cellular fluid volumes. These predictive equations were correlated with standard body fluid dilution reference methods. The result was an accuracy of ±0.64% for total body water, ±0.17% for plasma volume, ±1.91% for extra-cellular fluid, and ±0.57% for intra-cellular fluid compartments. Less than 5 min was required for all of the measurements and determinations. Therefore, it appears that an accurate measurement of body fluid distribution can be performed on horses using a fast, easy, non-invasive, inexpensive BIA method.     

Quantitative ultrastructural anatomy of esophagus in different regions in the horse: effects of alternate methods of tissue processing

A qualitative and quantitative study was made of 2 adult horses to determine the effect of specimen preparation and the influence of anatomically separate regions on the microscopic structure of the esophagus. The effects of 3 fixative solutions (formalin, Bouin's fixative and Karnovsky's fixative) and 4 histologic staining procedures (hematoxylin and eosin, Gomori's trichrome, Laidlow's reticulum, and orcein-elastin) were compared. Density-dependent image analysis was used to compare the relative contributions of elastic and reticular fibers in esophageal connective tissue for each region. Cross-sectional dimensions of the tissue layers which comprise the esophageal wall were obtained by light microscopic measurement and were compared for each region. The mucosal surface structure was examined by scanning electron microscopy. Our results indicate that specimen preparation has significant effects on the structural integrity of esophageal tissues. Bouin's fixative was inferior to the other fixatives in this regard. Furthermore, there are extensive regional differences in esophageal microstructure. Formalin-fixed tissues stained with either hematoxylin and eosin or Gomori's trichrome appear the most suitable for routine quantitative morphologic studies of the equine esophagus.