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.     

Horse vision and an explanation for the visual behaviour originally explained by the 'ramp retina'.

Here we provide confirmation that the 'ramp retina' of the horse, once thought to result in head rotating visual behaviour, does not exist. We found a 9% variation in axial length of the eye between the streak region and the dorsal periphery. However, the difference was in the opposite direction to that proposed for the 'ramp retina'. Furthermore, acuity in the narrow, intense visual streak in the inferior retina is 16.5 cycles per degree compared with 2.7 cycles per degree in the periphery. Therefore, it is improbable that the horse rotates its head to focus onto the peripheral retina. Rather, the horse rotates the nose up high to observe distant objects because binocular overlap is oriented down the nose, with a blind area directly in front of the forehead.     

Normal structure and age-related changes of the equine retina

Investigations of the pathophysiology of ocular diseases require a detailed knowledge of the microanatomy of the eye. The available information is still inadequate for the equine retina despite the importance of eye diseases in equine medicine. Here we provide a comprehensive analysis of the histologic features of the horse eye as a reference for future studies. Thirty normal eyes of 15 healthy horses were examined immediately after slaughter. The retina of the horse differs considerably in the degree and quantity of neurons and glial elements as well as in vascular patterns compared to the retina of other domestic animals. Morphometric analysis revealed that the thickness of the retina varies between 80 microm at the ora serrata and 250 microm medial to the optic disc. Approximately 90% of the equine retina is comparatively thin (< 130 microm). This is a physiologic response to the distance that oxygen can diffuse in avascular retina. Ganglion cells form a single layer in all parts of the retina. The majority of ganglion cells are very large Nissl-positive cells. Small Nissl-negative ganglion cells are less abundant. A high ganglion cell density is found only in the central area. Vascularization is virtually absent from the retina with the exception of a narrow strip around the disc of the optic nerve, as revealed by lectin histochemistry. Light microscopy of the eyes of older horses repeatedly revealed cystoid degenerations in the retina adjacent to the pars plana of the ciliary body, as well as a destruction of the regular layering of the peripheral region of the retina.     

Topography of ganglion cells in the retina of the horse

Topography of ganglion cells in the retina of the horse (Thoroughbred) was analyzed in the wholemount retina stained with cresyl violet, and a total number of ganglion cells were estimated. Sizes of ganglion cells were also measured and size spectra were analysed. The main results showed that: (1) a common point in 4 wholemount retinae, based on cell densities and retinal locations, was that a retina could be divided into 5 regions, namely visual streak, nasal, temporal, dorsal and ventral region to the visual streak. A maximum cell density of 4,000 cells/mm2 was found in the visual streak. And a total number of ganglion cells was estimated in a range of 398 x 10(3) - 469 x 10(3), with a mean of 441 x 10(3) +/- 31 x 10(3) (n=4). (2) cell sizes were measured as the mean lengths of the major and minor axes of the somas, and were in a range of 5-53.8 microm. The lowest mean diameter was 14.0 microm (+/- 3.7) in the visual streak and the highest was 25.9 microm (+/- 7.6) in the ventral region. Cell size spectra were unimodal and positively skewed. It is expected that these analyses will provide an anatomical and physiological background for further study of the visual system in the horse.     

Angiostatin and integrin alphavbeta3 in the feline, bovine, canine, equine, porcine and murine retina and cornea

PURPOSE: Angiogenesis is tightly controlled in the ocular tissues of domestic animals but its mechanisms are not fully understood. This is largely because of insufficient data on the expression of molecules that impact angiogenesis. Because angiostatin and one of its receptors integrin alphavbeta3 inhibit and promote angiogenesis, respectively, we hypothesized that the normal retina and cornea of domestic animals would express angiostatin but not integrin alphavbeta3. PROCEDURE: Normal eyes of the cat, cow, dog, horse, pig and rat were evaluated for angiostatin and integrin alphavbeta3 by light and electron immunocytochemistry and estern blots. RESULTS: Angiostatin was detected in the corneal epithelium of the cat, dog, horse, pig and rat, but was not found in cow corneal epithelium. Angiostatin was localized in the nerve fiber layer, ganglion cell layer, inner and outer plexiform layers, and the photoreceptor layer of the cat, cow, dog and rat. Horse and pig retinas showed additional staining in the matrix of the inner nuclear layer. Immunogold electron microscopy further confirmed angiostatin in cat retina. Western blots showed angiostatin in corneal and retinal homogenates. Integrin alphavbeta3 was absent in cornea and retina of all the species studied. CONCLUSION: These data show that angiostatin, an inhibitor of angiogenesis, is present while integrin alphavbeta3, which promotes angiogenesis, is absent in normal cornea and retina of the domestic animals in this study with the exception being angiostatin absence in cow corneal epithelium. Therefore, angiostatin may contribute to the anti-angiogenic environment in the normal domestic animal eye while its absence in the cow may contribute to greater propensity for corneal vascularization. Because integrin alphavbeta3 is one of the receptors for angiostatin, its absence may prevent angiostatin from killing normal retinal and corneal cells.     

Molecular genetic studies of the "Einsiedler" horse population

In this study it was investigated whether the "Einsiedler" warmblood horse, a historically old horse population from central Switzerland (Abbey of Einsiedeln), is distinguishable from micellaneous horse breeds, using molecular genetic techniques. The breeding history of Einsiedler horses is characterised by systematic line breeding through the dams. Therefore, two Einsiedler dam lines (N = 28), going back to the middle of the 19th century according to pedigree entries, were the focus of the survey. Random samples of diverse warmblood horse populations, but also samples from more distinct types of horse breeds, served as comparison populations (N = 52).Variation in the mitochondrial genome appeared to be only partially informative to demarcate the studied horses, as horses of distinct breeds may share identical mtDNA sequence fragments. Both dam lines revealed haplotypes commonly found in Iberian horse breeds. This is to take as an indication on the genetic origin of Einsiedler horses. Furthermore, the Klima dam line held a homologous mtDNA sequence fragment with E. ferus przewalskii. Therefore, this seems to be a phylogenetically old haplotype. The analysis of microsatellite loci revealed that horses from the two Einsiedler dam lines were in fact distinguishable from more distinct types of horses, but not from closely related European warmblood horse breeds and English thoroughbred.     

Direct current stimulation of bone production in the horse: preliminary study with a "gap healing" model

The effect of a 20-microA direct-current implantable bone growth stimulator (BGS) on bone production with a "gap healing" model in the horse was evaluated. The right and left 4th metatarsal bones (Mt-4) were used in 7 adult horses to create the "gap healing" model. A 4-mm section of the Mt-4 bone was resected bilaterally in each horse. The BGS was surgically placed into the 7 left Mt-4 defects. The 7 right Mt-4 defects served as controls. Six horses survived the 16-week experimental period. Signs of pain, decreased range of limb motion, or lameness was not observed in any animal during the 16 weeks. None of the animals showed complete healing radiographically. Four stimulated sites showed less periosteal reaction and 2 showed greater reaction than the 6 controls. The greatest amount of periosteal reaction or bone resorption was seen around the screws and plates in both groups. Uptakes of 99mTc-MDP in counts/pixel for control sites and stimulated sites were 7.90 and 8.25 in the "gap defect" and 5.19 and 5.06 in the areas adjacent to the gap defect. The ratio of uptake between the gap defect and adjacent area was 1.5 and 1.58 respectively. Biocompatability of the BGS was excellent; however, 1 horse had a broken cathode wire 5 cm from the generator capsule at 6 weeks. All polyethylene cathode sheaths were fluid filled at 16 weeks. The average mineralization rates were 1.57 +/- 0.34, 1.71 +/- 0.28 mm/day and bone formation activity was 0.0182 +/- 0.171, and 0.0168 +/- 0.0149 mm2/day for control limbs and stimulated limbs, respectively. There was no significant difference between groups in any of the histomorphometric values measured. Direct current (20 microA) did not increase bone production in this experiment. Methods to objectively evaluate electrically induced osteogenesis and a "gap defect" model for BGS research on the horse are discussed. The results provide a basis for additional research on electrical stimulation of fractures in the horse and for dose-response studies.     

Anthelmintic resistance in cyathostomins of brood horses in Ukraine and influence of anthelmintic treatments on strongylid community structure

In 2004-2006, 322 brood horses from 11 horse farms were examined using the faecal egg count reduction test (FECRT) to determine the presence and distribution of anthelmintic resistance in strongylids in Ukraine. The anthelmintic drugs "Albendazole-7.5" (7.5mg of albenazole, Ukraine) at a dose of 5mg per kg body weight and "Univerm" (0.2% aversectin C, Russia) at a dose of 0.5mg per kg body weight were used. Seventy-one horses from six farms were examined in vivo to investigate the influence of anthelmintic treatment on the gastrointestinal strongylid community structure. Horses were treated with anthelmintics; faecal sampling (200 g in each sample) for strongylid expulsion was performed 24, 36, 48 and 60 h after treatment; and all strongylids expelled (25,292 specimens) were collected and identified. Fourteen horses from the Dubrovsky horse farm were also examined to determine the benzimidazole-resistant cyathostomin species; 5208 specimens of benzimidazole-resistant cyathostomins were collected and identified. According to the FECRT data, benzimidazole resistance in strongylids was observed only at the Dubrovsky horse farm (FECRT=68.7%). No resistance to macrocyclic lactones in strongylids or in Parascaris equorum was observed. Twenty-nine strongylid species were found in horses from six horse farms. The number of species per horse ranged from 4-9 (5.8+/-1.5) to 10-20 (14.4+/-2.9) and depended on horse anthelmintic treatment strategies. From 4 to 13 strongylid species predominated (prevalence>66.7%) in the strongylid community. Eleven cyathostomin species (Cylicocyclus nassatus, C. ashworthi, C. leptostomum, Cyathostomum catinatum, C. pateratum, Cylicostephanus calicatus, C. longibursatus, C. goldi, C. minutus, Coronocyclus coronatus and C. labiatus) were found to be resistant to benzimidazoles at the Dubrovsky horse farm. Ten of these were the dominant species in the strongylid community; only C. labiatus was a rare species (prevalence 29.4%). Species richness and species diversity were significantly higher in horses from farms without treatment or with occasional treatments than from farms with regular treatments. The shape of the prevalence frequency distribution of strongylid species from farms with regular treatments was bimodal ("core" and "satellite" mode). This distribution was multimodal at farms without treatment or with occasional anthelmintic treatments. The results of the current study indicated the possibility of the further spread of anthelmintic resistance on horse farms in Ukraine and the necessity of monitoring the development of resistance in horse parasitic nematodes.     

Fine structure of the retinal pigment epithelium, Bruch's membrane and choriocapillaris in the horse

The fine structure of the retinal pigment epithelium (RPE), Bruch's membrane and choriocapillaris was investigated by light and transmission electron microscopy in both the tapetal and non-tapetal fundus of the horse eye. In all locations, the RPE consisted of a single layer of low cuboidal cells. The epithelial cells were joined laterally by apically located tight junctions. These cells displayed numerous basal infoldings and abundant thin apical processes which enclosed the rod outer segments. The epithelial cell nuclei were large and located basally. Within the epithelial cells, smooth endoplasmic reticulum was very abundant, while rough endoplasmic reticulum was scarce, polysomes and mitochondria, which often display a ring-shaped structure, were abundant. Melanosomes were abundant in the non-tapetal area but absent in the tapetal area. Bruch's membrane was pentalaminate throughout the retina. The endothelium of the choriocapillaris was heavily fenestrated.     

Retinal vascular patterns in domestic animals

In this paper a morphological study of the retinal vascular patterns in various species of domestic animals is reported. A classification of these patterns into four well-defined groups is described. In the domestic ruminants, pigs and carnivores the retina contains a compact plexus of blood vessels located in the major part of the light-sensitive portion of the retina (euangiotic or holangiotic pattern). In other domestic animals blood vessels are present only in a smaller part of the retina. In the rabbit, vessels are confined to a broad horizontal band coincident with the area of dispersion of the myelinated nerve fibres. The larger of these vessels are readily visible macroscopically (merangiotic pattern). In the horse and the guinea pig the retinal blood vessels are minute and restricted to the direct neighbourhood of the optic disc (paurangiotic pattern). The avian retina is completely avascular (anangiotic pattern), but a densely vascularised pecten oculi is attached to the linear optic nerve head and protrudes far into the inferior part of the vitreous body.     

The presence of heat-labile factors interfering with binding analysis of fibrinogen with ferritin in horse plasma

Background

Horse fibrinogen has been identified as a plasma specific ferritin-binding protein. There are two ways in the binding of ferritin-binding protein with ferritin: one is direct binding and the other is indirect binding which is heme-mediated. The aim of this study was to analyze the binding between horse fibrinogen and ferritin.

Findings

Although fibrinogen in horse plasma did not show the binding to ferritin coated on the plate wells, after following heat-treatment (60°C, 30 min) of horse plasma, plasma fibrinogen as well as purified horse fibrinogen bound to plates coated with horse spleen ferritin, but not with its apoferritin which lost heme as well as iron after the treatment of reducing reagent. Binding of purified or plasma fibrinogen to ferritin was inhibited by hemin and Sn-protoporphyrin IX (Sn-PPIX), but not by PPIX or Zn-PPIX.

Conclusions

Heat-treatment of horse plasma enabled plasma fibrinogen to bind to plate well coated with holo-ferritin. From the binding analysis of fibrinogen and ferritin, it is suggested that horse fibrinogen recognized iron or tin in complexed with the heme- or the hemin-ring, and also suggest that some fibrinogens circulate in the form of a complex with ferritin and/or heat-labile factors which inhibit the binding of fibrinogen with ferritin.     

Ganglionic cell distribution in the central area of the beagle dog retina

The distribution of ganglionic cells in the central area of the Beagle dog retina was studied by means of light microscopy in thirteen whole-mount preparations stained with cresyl violet. The central area was composed of two parts termed "the round central area" and the "horizontal streak"; in some specimens the former and in other the latter part was more prominent. The highest concentration of ganglionic cells occured in the round central area (about 7.100 cells/mm²). In other regions of the central area the cells were less numerous and the fewest number was found outside the central area. The size categories of ganglionic cells in the Beagle retina are comparable with those observed in other species.     

Aspects of the owning/keeping and disposal of horses, and how these relate to equine health/welfare in Ireland

Background

Ireland has long been renowned as a major centre for the breeding, rearing and keeping of horses. Since 2007, however, there has been increasing concern for horse health and welfare standards, and links between these concerns and the structures, governance and funding of the Irish equine industries have been reported. This paper addresses two central issues: firstly the local governance of, trade in and disposal of unwanted horses; and secondly mechanisms employed to improve standards of care given to horses owned by certain communities.

Method

Primary information was gathered through visits to horse pounds run by and on behalf of Local Authorities, to social horse projects, to horse dealer yards, ferry ports, horse slaughter plants and knackeries.

Results

The approach adopted by members of a given group, e.g. ferry ports, is described and differences are highlighted, for example in how different Local Authorities implement the Control of Horses Act of 1986, and how the choice, for example, of disposal route affects the standard of animal welfare.

Conclusions

There is a pressing need for a more centrally mandated and uniformly applied system of governance to safeguard the health and promote the keeping of horses to a higher welfare standard in Ireland. Fundamental to an understanding of why there is insufficient oversight of the keeping and proper disposal of horses is the lack of a comprehensive, integrated system for the registration, identification and tracing of equidae in Ireland.     

Chronology of horseshoes found in Switzerland

The horse was domesticated by a people in Central Asia. However, it is not known where and who invented horse shoeing, an invention that increased the possibilities for using horses. The word "horseshoe" was first mentioned in a document from 826 p.c. Up to now an archaeologist couldn't date an excavated horseshoe. I succeeded now to establish a chronology for horseshoes found in Switzerland. The first realisation is that in the course of more than thousand years of horse shoeing history three basic forms followed each other. Moreover, the existing horseshoes can be allocated to periodical types, each in use for approximately a quarter of a century.     

Gastrointestinal stromal tumour and hypoglycemia in a Fjord pony: Case report

Background

Neoplasia may cause hypoglycemia in different species including the horse, but hypoglycemia has not previously been reported in the horse associated with gastrointestinal stromal tumours.

Case presentation

A case of a gastrointestinal stromal tumour in a Fjord pony with severe recurrent hypoglycemia is presented. The mechanism causing the hypoglycemia was not established.

Conclusion

This case indicates that a gastrointestinal stromal tumour may cause hypoglycemia also in the horse.     

Agammaglobulinemia in a horse with evidence of functional T lymphocytes.

Agammaglobulinemia was diagnosed in a 1-year-old Thoroughbred horse on the basis of the following observations: (1) absence of serum immunoglobulins M, A, and G(T); (2) small amounts of serum immunoglobulin G (16 mg/100 ml); (3) absence of specific antibody in the serum of the horse following immunization and challenge exposure to 2 antigens; (4) absence of plasma cells, primary follicles, and germinal centers in a lymph node removed after antigenic stimulation; (5) absence of "natural" serum antibodies to rabbit-erythrocytes which were easily detectable in age-matched control horse serums; and (6) increased susceptibility to infections. There was evidence of functional cell-mediated immunity which included a skin response to injected phytolectins, skin response to antigen challenge following sensitization, and in vitro proliferative response of lymph node cells to phytohemagglutinin. An intact cell-mediated immune response was also supported by the observation that the horse lived to 17 months of age without antibody production, whereas horses with an absence of both antibody production and cell-mediated immunity (combined immunodeficiency) die by 4 months of age without immunologic intervention. The known features of agammaglobulinemia in this horse are similar to those in sex-linked agammaglobulinemia in persons and are unique among the immunodeficiences described in other animals.     

An equine cryptorchid with testicular and ovarian tissues.

Cytogenetic and histological studies were carried out on an intersex horse which was diagnosed clinically as a cryptorchid. Surgery confirmed the horse to be a bilateral abdominal cryptorchid and histological examination revealed ovarian tissue associated with the left epididymis. Chromosome analysis of cultured cells from testicular tissue, ovarian tissue and skin revealed 64-XX and 64-XY make-up, the left gonad containing a greater preponderance of XX cells over XY cells. The external characteristics and behaviour of the horse were indistinguishable from that of a "routine" cryptorchid. Other cases of equine intersexes are reviewed and theories for the discrepancies between genetic, gonadal and phenotypic sex are discussed.     

Scintigraphic Appearance of the Dorsal Cortex of the Third Metacarpus and Third Metatarsus in the Horse

The lack of "standard uptake pattern" to refer to when interpreting scintigraphic images poses a problem to anyone working with skeletal scintigraphy. An article written by Koblik et al. stated that in the normal scintigraphic image of older horses, the dorsal cortex of the metacarpus and metatarsus is not identifiable, whereas it can be identified in younger horses. In this retrospective study we evaluated the association between the age of a horse and visibility of the dorsal cortex of the metacarpus and metatarsus. We found that dorsal cortical uptake can be seen in the normal adult horse. The population in the study consisted of 139 horses scanned under general anesthesia in lateral recumbency. Scintigraphic images of 202 limbs were examined visually and by using a profile image tool. In almost all limbs the dorsal cortex was identifiable (82%); only 7% of the dorsal cortices were not identifiable; and 11% of the scintigraphic images were not interpretable.     

Pancarpal Arthrodesis for Treatment of Carpal Collapse in the Adult Horse A Report of Two Cases

Two mares with multiple carpal bone fractures, malarticulation and degenerative joint disease were successfully treated with unilateral arthrodeses of the antebrachiocarpal, middle carpal, and carpometacarpal joints. Arthrodesis was achieved by removal of articular cartilage, autogenous cancellous bone graft, and double dynamic compression plating. In one horse, wedge ostectomy corrected the severe, acquired angular limb deformity. External coaptation supplemented the internal fixation. Postoperative complications were limited to cast sores that healed with treatment, although protracted in one horse. Both mares adapted to the "peg leg" condition and ambulated without pain by 3 months. The first mare treated survived for at least 2 1/2 years after surgery and has had at least one foal. The most recently treated mare will be bred next year.     

Mitochondrial Profiles of the East Bulgarian and the Pleven Horse Breeds

It is well known that horse breeding in Bulgaria is a cultural heritage in Bulgaria, dating from prehistoric and historic times. Until now, molecular data on Bulgarian horses from the plain regions of the country were not available. Therefore, for the first time, we have collected genetic information about some modern horse breeds from the plain regions in Bulgaria. A total of 50 horses originating from different families from two different breeds were investigated: the first one was the Pleven horse (n = 11, breeding in the Danubian Plain), and the second one was the East Bulgarian horse breed (n = 39, Shumen district). These breeds were genotyped according to the mitochondrial D-loop region. The results showed that the Pleven horse particularly carries the European haplogroup L (45.45%), followed by the Middle East haplogroup C (27.27%). In contrast to the Pleven horse, the East Bulgarian horse breed revealed almost equal frequencies of the European specific haplogroup L (33.33%) and the Central Asiatic haplogroup Q (35.90%). Analyses of these two horse breeds revealed a specific genetic profile, but it is obvious that the East Bulgarian horse showed an unusual, mixed profile—a massive admixture with the Asiatic-type haplogroup Q and a high haplogroup diversity. In conclusion, the differences in genetic structure of the two plain horse breeds may be explained with the various horse breeds involved in their formation.