Identification of surface morphologic changes in the mitral valve leaflets and chordae tendineae of dogs with myxomatous degeneration

OBJECTIVE: To describe structural changes in the left atrioventricular (mitral) valve complex of dogs with endocardiosis by use of scanning electron microscopy. ANIMALS: 5 clinically normal dogs and 4 dogs with mitral valve endocardiosis. PROCEDURE: The mitral valve complex from each dog was fixed and prepared for examination via scanning electron microscopy. Findings in valves from clinically normal and affected dogs were compared to identify surface changes associated with endocardiosis. RESULTS: Compared with findings in valves from clinically normal dogs, endocardiosis-affected mitral valve complexes had several morphologic abnormalities. Tissue swelling on the edge of valve leaflets, chordae tendineae, and the chordal-papillary muscle junction was evident. Damage to the valve complex endothelium was unevenly distributed; in some areas, denudation of endothelial cells had exposed the basement membrane or subendothelial valve collagen matrix. This damage was most noticeable on the leaflet edges and extended more to the ventricular aspect of the valve than the atrial side. Cell loss also extended to the chordae tendineae but was less apparent at the chordal-papillary muscle junction. The remaining endothelial cells on affected valves were arranged in less-ordered rows and had more plasmalemmal microappendages, compared with cells on unaffected valves. CONCLUSIONS AND CLINICAL RELEVANCE: Morphologic changes associated with mitral valve endocardiosis in dogs were similar to those observed in humans with mitral valve prolapse. In dogs with mitral valve endocardiosis, gross changes in the valve complex may affect hemodynamics in the heart; alterations in the leaflet and chordal endothelium may contribute to pathogenesis of this disease.     

A new and unusual species of Parachiton (Mollusca: Polyplacophora) from South Africa

A new species of the genus Parachiton, P. hodgsoni n.sp., which inhabits shallow water off the South African coast, is described. The radula is unusual for the genus Parachiton. It consists of about 160 transverse rows of very small mature teeth of which the major lateral teeth have very narrow tridentate cusps. The intermediate valves are granulose with the granules arranged in widely spaced, longitudinal rows on the central area of the valves. There are 20–31 longitudinal rows on the central areas of the intermediate valves. Variability in the morphology of the tail valve is shown.     

Clinical, echocardiographic, and Doppler imaging characteristics of mitral valve stenosis in two dogs.

Mitral stenosis was diagnosed noninvasively by echocardiography and Doppler imaging in 2 Bull Terriers. Two-dimensional echocardiography revealed severe atrial and moderate left ventricular dilatation; severely reduced mitral valve opening excursion; doming of the cranial mitral valve leaflet into the left ventricle during diastole; thickened, nodular cranial mitral valve leaflets; and reduced mitral valve orifice. M-mode echocardiographic findings additionally indicated greatly diminished mitral valve E to F slope and abnormal caudal mitral valve leaflet motion. Color flow Doppler imaging revealed bright bursts of color with aliasing originating from the stenotic mitral valve orifice, extending into the left atrium during systole, and into the left atrium during diastole. Spectral Doppler recordings revealed transvalvular mitral valve gradients and prolonged pressure half-times. Necropsy performed on 1 dog revealed extremely thickened, nodular, and stiff mitral valves with short, thickened, and fused chordae tendineae. The diagnosis of mitral valve stenosis was easily facilitated with diagnostic ultrasonography.     

Ultrastructural morphologic evaluation of the phenotype of valvular interstitial cells in dogs with myxomatous degeneration of the mitral valve

OBJECTIVE: To evaluate morphologic changes in valvular interstitial cells of dogs and to find evidence for disease-associated phenotypic changes in these cells. ANIMALS: 5 clinically normal dogs and 5 dogs with severe mitral valve endocardiosis. PROCEDURE: Mitral valve leaflets were evaluated by use of transmission electron microscopy. Differences in cell type and cell location were identified. RESULTS: A change in cell type toward a myofibroblast or smooth muscle cell phenotype was detected, with the smooth muscle cell type being most common. These cells had long amorphous cytoplasmic extensions, fibrillar cytoplasm, incomplete basal lamina, few mitochondria, and eccentrically placed nuclei but lacked smooth endoplasmic reticulum or Golgi complexes. Remaining valvular interstitial cells had heterochromatic nuclei and produced only minimal quantities of collagen. Compared with normal valves, myxomatous valves ha many interstitial-like cells located adjacent to the endothelium. Deeper within the abnormal valves, cells with a heterogenous phenotype formed groupings that appeared to be anchored to adjacent collagen. CONCLUSIONS AND CLINICAL RELEVANCE: Myxomatous degeneration of the mitral valve in dogs is associated with phenotypic alteration, changing from an interstitial to a mixed myofibroblast or smooth muscle cell phenotype. A closer association between interstitial cells and the endothelium is evident in diseased valves. In response to the disease process, valvular interstitial cells of dogs appear to change toward a smooth muscle phenotype, possibly in an attempt to maintain valve tone and mechanical function.     

Extracorporeal Bypass of the Right Ventricle and Pulmonary Trunk Using Autogenous Lungs for Oxygenation in the Dog

A technique for right heart bypass using autogenous lungs for oxygenation was evaluated. Venous blood was collected by a catheter placed in the right atrium and caudal vena cava. The blood then passed through the extracorporeal unit and was pumped into the pulmonary circulation. This technique allowed direct surgical exposure of the conus arteriosus, pulmonary valve, and pulmonary trunk. The surgical procedure, plasma hemoglobin levels, platelet and leukocyte counts, arterial blood gas, serum proteins, electrocardiogram, and postmortem findings were evaluated. Although there were significant alterations in many of these parameters, seven of eight dogs were successfully perfused and recovered rapidly from the surgery. One dog died of complications arising from inadequate venous return before bypass was established. On the first postoperative day, the survivors were ambulatory, eating, and having normal bowel function. Right heart bypass may be an effective method to approach lesions involving the conus arteriosus, pulmonary valve, and pulmonary trunk.     

Structural and Histochemical Studies on the Teleostean Bulbus Arteriosus

The structure and histochemical properties of the bulbus arteriosus in two species from an evolutionary old teleost family, Characidae, and in three modern teleosts, family Cichlidae, are described. The bulbar wall was composed of an outer layer, a middle layer and a strongly folded inner layer covered by a thick, granule‐rich endothelial cell layer towards the lumen. One of the cichlid species (Thorichthys meeki) was injected intraperitoneally with horse ferritin; the endothelial cell layer of the heart atrium and ventricle displayed high ability to endocytose ferritin particles from the blood stream, but the corresponding layer in the bulbus arteriosus displayed no such uptake. This finding suggests that the bulbar endothelial cell layer plays no scavenger or immunological blood cleansing roles in this species. The bulbar endothelial cell granules were strongly coloured by periodic acid–Schiff (PAS) in the present cichlids, but weakly coloured by PAS in the present characids. These cell layers were uncoloured by alkaline carmine in ethanol in both cichlids and characids. The negative carmine test combined with a positive PAS test for the bulbar endothelial cell layer in the present cichlids indicates that these cells contain only small amounts of polysaccharides. The weak PAS‐colouring for the bulbar endothelial cell layer in characids indicates a very low content of sugars in these cells. These findings together with the fact that this cell layer in the present cichlids and characids was nearly uncoloured when treated with orcein, Heidenhain’s Azan or Schmorl’s solutions for elastic materials suggest that the bulbar endothelial granules do not play any role in the blood cleansing or in the rebuilding or maintenance of the ground substance or elastic material in the bulbar wall. Probably, the granules in the bulbar endothelial cell layer in the present species contain mainly proteins, connected to some PAS‐positive polysaccharides to enhance their solubility.     

ECHOCARDIOGRAPHIC ASSESSMENT OF 537 DOGS WITH MITRAL VALVE PROLAPSE AND LEAFLET INVOLVEMENT

In this work we investigated which mitral valve leaflet was most often involved in mitral valve prolapse with degenerative mitral valve disease and whether there was an association with breed, age, gender, or weight. Five hundred and thirty-seven dogs with mitral valve prolapse-degenerative mitral valve disease were assessed; the cross-breed dog was the most represented breed (248 dogs, 46.2%). Mitral valve prolapse was more common in male dogs, and the average age was 11.3±2.8 years. Prolapse of the anterior leaflet was present in 48.4% of dogs, prolapse of the the posterior leaflet in 7.1%, and bileaflet prolapse was present in 44.5%; this distribution is different than that typically found in humans. There was a significant correlation between severity of mitral regurgitation and severity of mitral valve prolapse or ISACHC class, and between severity of mitral valve prolapse and ISACHC class. There was no relationship between the particular affected leaflet(s) and severity of mitral regurgitation, severity of mitral valve prolapse, or ISACHC class. Our findings suggest that the susceptibility to the mitral valve prolapse-degenerative mitral valve disease is not confined to a specific breeds and that the specific leaflet prolapsing is different in dogs compared with humans.     

No expression of angiotensin II receptors and angiotensin-converting enzyme in myxomatous canine mitral valve leaflets. An autoradiographic study

The renin-angiotensin system, including angiotensin (Ang) II and angiotensin-converting enzyme (ACE), plays an important role in cardiac fibrous tissue formation. Since changes in valvular collagen are a central part of myxomatous mitral valve disease in the dog, we speculated that Ang II and ACE might play a role in the pathogenesis of this disease. In 10 mitral valves, five with and five without clear myxomatous changes, the presence and distribution of Ang II receptors and ACE was examined autoradiographically, using 125I-Ang II and 125I-lisinopril, respectively. At postmortem examination, diseased valves were taken from old dogs, control valves from young adult dogs. No significant level of Ang II and lisinopril binding was found in normal as well as diseased valve leaflets. Equally low, insignificant levels of 125I-Ang II binding were found in the myocardium of dogs with and without valvular disease. No significant level of myocardial 125I-lisinopril binding was found. The lack of autoradiographic evidence of Ang II receptors and ACE in normal and diseased canine mitral valve leaflets suggests that the renin-angiotensin system does not play a major role in the pathogenesis of the valvular changes.     

Structural remodelling of the heart valves extracellular matrix during embryo development

Alterations in heart valve development represent more than 20% of congenital cardiovascular malformations. Most of the functional properties of heart valves depend on extracellular matrix. Despite its relevance, little is known about fibrillar components on developing stages. Our objective is to define histological changes on valves fibrillar components in late embryonic development of Mus musculus. We found type III collagen as the predominant fibre type in the ECM in prenatal stages followed by a switch to a type I predominance for postnatal ages. The change in fibrillar components is necessary to support the normal mechanical function of adult heart valves.     

Dynamic obstruction of the left ventricular outflow tract in four young dogs

Four young dogs presented for evaluation of left-sided systolic heart murmurs all showed echocardiographic changes consistent with dynamic left ventricular outflow tract (LVOT) obstruction and subjective evidence of concentric left ventricular hypertrophy. In three of the dogs, abnormal mitral valve apparatus and systolic anterior motion of the anterior mitral valve leaflet with associated mitral insufficiency were also detected. All dogs were medicated with a beta1-adrenergic antagonist. Subsequent examinations showed that the dynamic LVOT obstruction and left ventricular concentric hypertrophy had almost completely resolved. Dynamic LVOT obstruction is a rare condition of young dogs of different breeds. The precise aetiology of the condition remains uncertain. Whether resolution of the outflow obstruction in these four cases was a consequence of treatment or due to changes in ventricular architecture brought about by ageing cannot be established.     

Pathologic and clinical features of infectious endocarditis

Infectious endocarditis is a systemic disease associated with high morbidity and mortality. Clinical recognition and effective management is challenging, but insights can be gleaned from relevant pathologic features. Risk factors include subaortic stenosis, possibly certain other congenital anomalies, and bacteremia. Auscultation can provide clues regarding valvular involvment, particularly when a diastolic left basilar murmur of aortic valve regurgitation is present. Aortic valve vegetations and insufficiency may also alter femoral arterial pulse characteristics. Echocardiography may facilitate diagnosis, particularly with aortic valve lesions, but may not be able to distinguish between small mitral valve vegetations and early chronic degenerative valve disease. Vegetative lesions develop along edges of valve closure on the ventricular aspect of the aortic valve and the atrial surface of atrioventricular valves. They may extend across valve leaflet, from valves to adjacent left atrial endocardium, interventricular or interatrial septum, or chordae tendineae. Vegetations can be friable and frequently embolize to spleen, kidney, and left ventricle — often before clinical recognition of the disease. Valvular insufficiency develops as a consequence of valvular vegetations, necrosis, perforation, or rupture of the chordae tendineae. Histopathologic appearance varies with respect to duration of disease and antimicrobial therapy. These factors influence the amount of necrotic material, blood clot, fibrin, and inflammatory cells which make up the vegetations. Bacteria are not always identified in valvular lesions, especially following antibacterial therapy, but may be detected in other organs. Common sequellae include congestive heart failure, sepsis, arrhythmias, and systemic organ infarction.     

Morphologic features of the aqueous humor drainage pathways in horses

The iridocorneal angle of the horse was anteriorly lined by a pectinate ligament, consisting of several layers of vertically oriented collagenous beams, that were interrupted by a network of extracellular spaces. The anterior chamber angle consisted mostly of a large uveal trabecular meshwork that was composed of widely-spaced, thick trabeculae. These trabeculae were attached in part posteriorly to extensions of the ciliary body musculature and anteriorly inserted into the prominent pectinate ligament. The corneoscleral trabecular meshwork was shaped hemiobovately and formed the external boundary of the iridocorneal angle. The extracellular spaces were smaller and more evenly spaced than those in the uveal trabecular meshwork. Uveal and corneoscleral trabecular meshwork beams were mostly composed of cores of collagen and elastin surrounded by several layers of basement membrane-like material and, in turn, were entirely enveloped by trabecular cells. An anterior narrow zone of broad corneoscleral trabeculae was composed of elastin-collagen cores surrounded by circular bundles of long-spacing collagen. The angular aqueous plexus was extensive, forming much of the external border of the outer corneoscleral trabecular meshwork. A dense zone was associated with the angular aqueous plexus, completely encompassing these aqueous outflow vessels. Posteriorly, the iridocorneal angle was confluent with a well-defined band of trabeculae that was situated between the ciliary body musculature and subjacent sclera and extended posteriorly to the anterior-most region of the choroid where these trabeculae tapered and ended. This region is referred to as the supraciliary trabecular meshwork and indicates a potentially extensive route for outflow of aqueous humor in the equine eye.     

Morphologic alterations in the anterior lens capsule of canine eyes with cataracts

OBJECTIVE: To examine the morphologic changes in the anterior lens capsule and lens epithelium of canine eyes with cataracts. SAMPLE POPULATION: Anterior lens capsules from the eyes of 25 dogs with cataracts and from an additional 10 canine globes with lenses subjectively assessed to be normal. PROCEDURE: Thickness of each anterior lens capsule was measured by use of a digital microscopic camera and imaging software. All 25 capsules from eyes with cataracts were submitted for light microscopy; 4 were also submitted for electron microscopy. RESULTS: Thickness of the anterior lens capsule increased with age for the normal lenses and the lenses with cataracts; the change with age was similar for both groups. Light microscopy revealed fibrous metaplasia of lens epithelial cells in 7 of 25 anterior lens capsules with focal thickenings of the posterior aspect of the capsule. Electron microscopy revealed deposition of collagen and basement membrane-like material by fibroblast-like cells. CONCLUSIONS: Results indicate that thickness of the anterior lens capsule in dogs increases with age and that this increase in thickness is not significantly different between normal lenses and lenses with cataracts. In addition, epithelial cells from lenses with cataracts may undergo metaplasia to form plaques composed of fibrous tissue and ectopic basement membrane produced by epithelial cells.     

Molecular species identification and population genetics of chondrichthyans in South Africa: current challenges,priorities and progress

Molecular genetic techniques, such as DNA barcoding and genotyping, are increasingly being used to assist with the conservation and management of chondrichthyans worldwide. Southern Africa is a shark biodiversity hotspot, with a large number of endemic species. According to the IUCN Red List, a quarter of South Africa's chondrichthyans are threatened with extinction. South Africa's commercial shark fisheries have increased over the last decade and there is a need to ensure sustainable utilisation and conservation of this fisheries resource. Here, we give an overview of the molecular techniques that are used to assist in the identification of species, cryptic speciation and possible interspecific hybridisation, as well as the assessment of population structure and reproductive behaviour of endemic and commercially important chondrichthyan species of southern Africa. We discuss the potential application of these techniques for management and conservation of several species affected by South African fisheries. Acquiring baseline barcode data of all chondrichthyans in southern African waters and assessing the population structure of exploited species on a local and greater regional scale are recommended as research priorities. Future prospects should also include high-throughput molecular marker development and investigation of intraspecies functional variation using next generation sequencing technology.     

Echocardiographic evaluation of equine aortic insufficiency

Echocardiographic evaluation of 23 horses with aortic insufficiency was performed, using M-mode (n = 23) and 2-dimensional real-time echocardiography (n = 14 of 23). Echocardiograms were evaluated for abnormalities of aortic and mitral valves and alterations in motion of these valves. Changes in left ventricular chamber size and function, as well as aortic root size, were evaluated. The presence of other cardiac disease was also evaluated. Horses with aortic insufficiency had significant increases (P less than 0.01) in mean values of left ventricular chamber size, aortic root diameter, and shortening fraction. Left ventricular free wall thickness also was significantly decreased (P less than 0.01). Valvular abnormalities were seen echocardiographically in all 23 horses. Eighteen horses with aortic insufficiency had thickened valves, whereas two horses had lesions associated with vegetative endocarditis. High-frequency vibrations of the septal leaflet of the mitral valve were noticed in all horses, whereas similar vibrations of the aortic valve were seen in six horses. The presence of a bounding arterial pulse correlated significantly (P less than 0.05) with increased left ventricular chamber size at end diastole and shortening fraction, indicating a marked left ventricular volume overload. A reliable diagnosis of aortic insufficiency can be made with the detection of bounding arterial pulses in concert with a holodiastolic decrescendo grade II to V/V murmur with maximal intensity over the aortic valve area, radiating toward the left cardiac apex.     

Fine Structure of the Bovine Tapetum Fibrosum

The fine structure of the tapetum lucidum of the domestic cow was studied by electron microscopy. In this ungulate the reflective layer is a tapetum fibrosum situated in the choroid and mainly restricted to the superior fundus. The tapetum is composed of a large array of fine extracellular collagen fibrils separated into lamellae by fibrocytes. The collagen fibrils display the normal cross-striations of native collagen, are 0.2 μm in diameter and are arranged in a hexagonal pattern with a center-to-center spacing of about 0.2 μm. The diameter and spacing of these fibrils is consistent with the constructive interference of reflected light. A vascular lamella adjacent to the retinal epithelial layer contains the choriocapillaris and some irregularly arranged collagen fibrils. The tapetum is pierced by blood vessels which supply the choriocapillaris. Over the tapetum the retinal epithelium is non-pigmented while in nontapetal regions this layer is pigmented. The choriocapillaris is not indented into the retinal epithelium as is the case in species with a tapetum cellulosum.