Scanning electron microscopy of vascular corrosion casts and histologic examination of pulmonary microvasculature in dogs with dirofilariosis

OBJECTIVE: To characterize structural changes in pulmonary vessels of dogs with dirofilariosis. ANIMALS: 8 dogs with dirofilariosis and 2 unaffected control dogs. PROCEDURE: Pulmonary artery pressure was measured in affected dogs, and dogs then were euthanatized. Scanning electron microscopy was used to examine vascular corrosion casts of pulmonary vasculature. Tissue sections of pulmonary vasculature were evaluated by use of histologic examination. RESULTS: Pulmonary artery pressure was higher in dogs with severely affected pulmonary vessels. In tissue sections, dilatation, as well as lesions in the tunica intima and proliferative lesions resulting in constriction or obstruction, were frequently observed in branches of the pulmonary artery. Numerous dilated bronchial arteries were observed around affected pulmonary arteries. Hyperplastic venous sphincters were observed in small pulmonary veins and venules. In corrosion casts, affected pulmonary lobar arteries had dilatation, pruning, abnormal tapering, constriction, and obstruction. In small arteries and arterioles, surface structures representing aneurisms and edema were seen. Bronchial arteries were well developed and extremely dilated, and they formed numerous anastomoses with pulmonary arteries at all levels, from the pulmonary trunk to peripheral vessels. Capillaries in the lungs were dilated with little structural change. Small pulmonary veins and venules had irregular annular constrictions that were caused by hyperplastic smooth muscle cells of venous sphincters. CONCLUSIONS AND CLINICAL RELEVANCE: Scanning electron microscopy of microvascular casts delineated links between the bronchial and pulmonary circulations in dogs with dirofilariosis. Results of scanning electron microscopy provided a structural explanation for the development of pulmonary circulatory disturbances and pulmonary hypertension in dogs affected by dirofilariosis.     

Idiopathic arterial medial calcification of the thoracic arteries in an adult horse

A 6-year-old, gelded, Paint horse displayed clinical signs of muscle wasting and limb stiffness for a 6-month period. The horse's clinical signs abated with corticosteroid therapy, but returned upon cessation of treatment. Upon necropsy, severe lesions of aortic thickening and aortic valve rigidity were observed. Histologically, the tunica media of the aorta, coronary arteries, and pulmonary arteries were expanded by foci of elastin fiber calcification and extracellular matrix with lacunae formation. The vascular lesions are comparative to what has been described as medial arterial calcification, seen in humans suffering from chronic renal failure or diabetes mellitus. No exposure to vitamin D-containing plants or feedstuff could be documented at the time of onset or during the period of clinical signs. The current case describes dramatic lesions of arterial medial calcification of the aorta, coronary, and pulmonary arteries of undetermined cause.     

Early arterial injury-induced myointimal proliferation in canine pulmonary arteries

Transmission electron microscopy was used to study the early vascular response induced by arterial damage with heartworm infection. The pulmonary arteries were examined in dogs 4 days after experimental transplantation of 6 to 8 Dirofilaria immitis adults. Evan's blue dye was given IV and followed in 60 minutes by perfusion fixation with 1% glutaraldehyde. Endothelial cell junctions were disrupted and rounded endothelial cells were observed. Macrophages and neutrophils adhered to abnormal endothelial cells. Focal areas had platelet aggregates adhered to exposed subendothelial structures. Platelets were activated and degranulated. Areas of the internal elastic lamina appeared to be disrupted, and smooth muscle cell processes from the tunica media extended through these regions of disruption. Smooth muscle cells were oriented toward the arterial lumen and some had migrated to the surface. These findings are compatible with the response to injury theory of myointimal proliferation.     

Histological and ultrastructural studies on the coronary artery of adult domestic dog (Canis familiaris)

The objective of this work was to study the histological structure of the dog's coronary artery by light and transmission electron microscope (TEM). The coronary artery consisted of three tunics: tunica intima, tunica media and tunica adventitia. The tunica intima consisted of endothelium rested directly on internal elastic lamina without the subendothelial connective tissue layer. The tunica media were composed of smooth muscle fibres interspersed with few elastic and collagen fibres. The tunica adventitia consisted of collagen and elastic fibres intermingled with fibroblast cells; it had vasa vasorum and nervi vasorum. Some histomorphometric measurements were performed and compared statistically. The ultrastructural study showed that the endothelial cells were communicated through complex junctions and characterised by filiform cytoplasmic processes passed through the opening of the underlying internal elastic membrane. The smooth muscle fibres of tunica media communicated with each other through cytoplasmic processes. The tunica adventitia showed minute non-myelinated nerve. This work revealed that the dog's coronary arteries are typical muscular arteries, which show little structural variations from that of other mammals.     

Immunohistochemical detection of apolipoprotein A-I and B-100 in canine atherosclerotic lesions

We attempt to determine and compare the localization of apolipoproteins (apo) apoA-I and B-100 in atherosclerotic lesions of canine aortas, coronary arteries, and the peripheral arteries, using immunohistochemical techniques. Histopathologically, atherosclerotic lesions were characterized by deposition of lipids and infiltration of lipid-laden foamy cells in the tunica intima and tunica media, sometimes forming fibrofatty plaques containing abundant sudanophilic and mineralized material. Canine apoA (CapoA)-I and canine apoB (CapoB)-100 immunopositive signals were simultaneously observed in mild and severe atherosclerotic lesions of the aorta, coronary arteries, splenic arteries, and renal arteries in the double-immunolabeled sections. Both CapoA-I and CapoB-100 positive signals were seen in the cytoplasm of endothelial cells, smooth muscle cells, and macrophages. The subendothelial space and extracellular matrix in the tunica intima and media were also positive. Neither CapoA-I nor CapoB-100 positive signals were seen in normal arteries. These findings closely resemble those of the localization of apoA-I and apoB-100 in human atherosclerotic lesions.     

Pulmonary hypertension as a cause of atrial fibrillation in young horses: four cases (1980-1989)

Four young horses of various breeds and suffering from atrial fibrillation died of heart failure. All had markedly high pulmonary arterial pressure, right-sided cardiomegaly, and lack of histologic lesions in the right atrium or pulmonary parenchyma. Three horses had hypertrophy and/or necrosis of the tunica media of the pulmonary vasculature. Clinical signs of disease, physiologic data, and pathologic findings indicated that these horses had primary pulmonary hypertension with secondary right-sided cardiac ventricular hypertrophy and dilatation, atrial fibrillation, and heart failure.     

Histopathological and enzyme histochemical observations on mast cells in pulmonary arterial lesion of dogs with Dirofilaria immitis infestation

Histopathological and enzyme histochemical observations were performed on mast cells in pulmonary arterial lesion of Dirofilaria immitis in dogs. The results showed that chymase- and tryptase-positive mast cells were diffusely present in the lesions, especially in the adventitia and proliferated intima. At 2 weeks after surgical worm transplantation, mast cells already appeared in the intima and media, and chymase-positive cells were dominant in the adventitia. Results of this study suggested a possibility that mast cells would be involved in the pathogenesis of pulmonary arterial lesion of dogs with Dirofilaria immitis infestation.     

Arterial calcification in race horses

Calcification of large arteries has been sporadically reported in horses. The pathogenesis is still unknown, but recent studies in humans suggest that this is a regulated biomineralizing process. This study surveyed the prevalence, distribution, and severity of vascular calcification in Thoroughbred and Standardbred racehorses. Histopathologic, ultrastructural imaging, and energy dispersive X-ray elemental analyses were used to examine the lesions. Calcification of the tunica media, predominantly the pulmonary artery, was found in 82% of horses (83/101). Young adult horses (mean [SD] age in years, 4.44 +/- 2.17) of both breeds and sexes were similarly affected. Lesions appeared as white-to-yellowish, hard, and gritty plaques of variable size. On microscopic examination, elastic fibers within the tunica media were thinned, fragmented, and calcified, and surrounded by dense collagen matrix. Elemental analysis showed distinct peaks for calcium and phosphorus, consistent with hydroxyapatite mineral. The frequent occurrence of calcification in the tunica media of large pulmonary arteries of young racing horses indicates the need to investigate its pathogenesis and potential clinical implications.     

Vascular lesions in nine Göttingen minipigs with thrombocytopenic purpura syndrome

Tissues from 9 G?ttingen minipigs, aged 7 weeks to 1 year, with clinically diagnosed thrombocytopenic purpura syndrome were examined microscopically. All pigs had a history of spontaneous cutaneous purpura that was generally accompanied by disseminated visceral hemorrhages. Hematologic abnormalities included anemia (8 out of 9 pigs) and thrombocytopenia (7 out of 9 pigs), with platelet counts consistently below 20,000/microl. Microscopically, degenerative vascular lesions with morphologic features of arteriosclerosis were present in all 9 pigs. Vascular lesions affected small- to medium-sized muscular arteries and arterioles in various organs and extraparenchymal tissues; vessels of the renal pelvis and coronary arteries were consistently involved. Microscopic lesions in small- to medium-sized muscular arteries consisted of neointimal proliferation, medial thickening, luminal stenosis, thrombosis, disruption and fragmentation of the internal elastic lamina, necrosis of the tunica media, and medial deposits of myxoid matrix material. Microscopic lesions in arterioles included concentric laminar thickening of vessel walls (onion-skin pattern), endothelial cell hypertrophy, smooth muscle cell vacuolation, necrosis of the tunica media, thrombosis, and partial to complete luminal stenosis. Arteritis and/or periarteritis were also noted in 4 out of 9 pigs. Additional microscopic lesions included membranoproliferative glomerulonephritis (3 out of 9), myocardial microinfarcts (4 out of 7), renal interstitial fibrosis (2 out of 9), extramedullary hematopoiesis (6 out of 9), and intracapillary hyaline thrombi (2 out of 9). Degenerative vascular lesions have not been previously described in G?ttingen minipigs with thrombocytopenic purpura syndrome. The etiopathogenesis of both the vascular lesions and thrombocytopenic purpura syndrome is currently unknown.     

The ultrastructure of Strongylus vulgaris-mediated equine chronic mesenteric arteritis

Cells found in the intima and media of the cranial mesenteric artery of a mature mare with chronic arteritis were identified as smooth muscle cells and occurred in association with collagen and elastin fibres. As no fibroblasts were demonstrable within these regions, the smooth muscle cells were the likely source of the extracellular matrix. In contrast, the abnormal adventitis from the same artery contained abundant fibroblasts which are considered to be the source of the adventitial collagen.     

AORTIC AND CARDIAC MINERALIZATION IN THE DOG

Aortic and cardiac mineralization was found in 21 of 3443 (0.61%) canine thoracic radiographs. In none of 786 feline thoracic radiographs reviewed were such lesions present. Mineralizations were superimposed on the ascending aorta (19 dogs) or on the caudal cardiac silhouette (2 dogs). In 2 of 4 dogs mineralization was identified echocardiographically dorsal to the aortic valve in close proximity to coronary arteries. Computed tomography confirmed mineralization of the aortic arch and root in 2 of 2 dogs. Necropsy and histopathologic examination in 1 dog revealed multiple nodular aortic tunica media calcifications with adjacent areas of degeneration. Lesions were significantly overrepresented in older dogs and in Rottweilers, and regarded as dystrophic calcification, caused either by age-related degenerative changes or chronic disease-related processes. There was no evidence of clinical significance attributed to the mineralization in any dog. Aortic and cardiac mineralization should be recognized as an incidental, non-significant finding in dogs of advanced age and differentiated from pleural and pulmonary structures.     

Pulmonary artery dissection causing haemothorax in a cat: potential role of Dirofilaria immitis infection and literature review

A 7-year-old male castrated domestic short-haired cat suddenly died. Gross examination revealed severe right-sided haemothorax with blood clots, four adult filarial nematodes in the blood clots and the caudal vena cava and haemorrhage dissecting into the tunica media of the right pulmonary artery. Histopathological investigation showed fibrosis of the tunica intima and disorganization/fragmentation of the elastic fibres accompanied by fibrous tissue deposition in the tunica media of both branches of pulmonary artery. Degenerative vasculopathy (intimal fibromuscular hyperplasia and medial hypertrophy/hyperplasia) involving pulmonary arteries was also observed. The polymerase chain reaction amplification and sequencing confirmed the identification of the parasite as Dirofilaria immitis. A diagnosis of pulmonary artery dissection with haemothorax and concomitant heartworm disease was formulated. Degenerative processes of the tunica media have been reported to cause pulmonary artery dissection in both humans and animals. Pulmonary artery remodelling induced by heartworms may be considered the underlying cause in the first case of feline pulmonary artery dissection, herein described.     

Histomorphologic and morphometric evaluation of the uterine horns in nulliparous and multiparous Beagles

OBJECTIVE: To macroscopically, histomorphologically, and morphometrically compare uterine tissues obtained from nulliparous and multiparous dogs. ANIMALS: 12 dogs constituting 2 homogenous groups (7 nulliparous Beagles and 5 multiparous Beagles, each of which had at least 7 pregnancies). PROCEDURE: Serum concentrations of progesterone and estradiol-17beta were determined. Samples of uterine tissues were fixed, embedded in paraffin, and cut into serial cross sections. Morphometric analysis was performed on systematically selected trichrome-stained sections. RESULTS: Mean absolute volume of the uterine wall did not differ between groups. Volume of blood vessels was significantly higher and relative mean value for myometrial connective tissue was significantly lower in the multiparous dogs. Arteries in the myometrium of multiparous dogs had pronounced thickening of the intima (ie, pregnancy sclerosis) and a concomitant thinning of the smooth muscle layer (tunica media). Furthermore, the elastica interna of these arteries appeared to be disintegrating and was highly and irregularly folded. Adventitia of the myometrial veins in multiparous dogs contained numerous layers of elastic fibers; however, only a few layers were observed in the adventitia of corresponding veins in nulliparous dogs. CONCLUSIONS AND CLINICAL RELEVANCE: In this study, we documented that in contrast to other species, the uterus of nonpregnant dogs does not increase in size and volume even after at least 7 pregnancies. Furthermore, sclerotic alterations of uterine arteries are proof of at least 1 pregnancy. Results of this study may be useful in the evaluation of uterine diseases.     

Treatment of canine dirofilariasis: pulmonary thromboembolism caused by thiacetarsamide--microscopic changes

Light and scanning electron microscopies were used to study the pulmonary embolism occurring in Dirofilaria immitis-infected dogs after treatment with thiacetarsamide. Lesions in control dogs (nontreated) which had been infected for 4 weeks with 9 Dirofilaria immitis adults were compared with lesions occurring in infected dogs at 2 weeks and at 4 weeks after they were treated with the adulticide. Extensive thromboembolism occurred in the caudal lobar pulmonary arteries of dogs at posttreatment weeks 2 and 4. Complicated villous proliferations were present at posttreatment week 2. The characteristic myointimal proliferation of dirofilariasis showed resolution in the large pulmonary arteries of the dogs at week 4. However, the caudal lobar pulmonary arterial and lung lesions were more severe in the later group. The pathophysiology of adulticide-induced thromboembolism and associated lung parenchymal changes were discussed.     

Relationship between pulmonary arterial pressure and lesions in the pulmonary arteries and parenchyma, and cardiac valves in canine dirofilariasis.

The relationship between pulmonary hypertension and lesions was examined in 41 dogs infested naturally with heartworms, which consisted of 28 cases with pulmonary heartworm disease and 13 cases with caval syndrome. Pulmonary arterial pressure (PAP) was measured before and 1 or 7 days after heartworm removal with a flexible alligator forceps. In these dogs, lesions were examined after the last measurement of PAP. The mean PAP was 28.2 +/- 16.0 mmHg (10.9 to 81.4 mmHg in range) at post-removal phase. Pulmonary arterial intimal lesions, pulmonary thromboemboli, pneumonic lesions, tricuspid valvular lesions and mitral valvular lesions were macroscopically recognized in 95, 59, 39, 54 and 56% of cases, respectively. These lesions were classified by severity and the relationship with PAP was examined by the multiple correlation analysis. The multiple coefficient correlation was found the highest between PAP and thromboemboli, followed by mitral valvular lesion, tricuspid valvular lesion, and pneumonic lesion. There was no significant correlation between PAP and intimal lesions. The coefficient of determination showed the highest value in thromboemboli when one variable was used, and increased only very slightly when a variable of thromboemboli was added to those of other lesions. The cases with high PAP had fresh thromboemboli in large pulmonary arteries. From these evidences, it was concluded that thromboemboli following natural death of heartworm was the most important factor causing an increase in PAP and developing clinical signs in canine heartworm disease.     

Cerebral arteriosclerosis in an aged captive polar bear (Ursus maritimus).

Cerebral arteriosclerosis was observed upon necropsy of a 36-yr-old female captive polar bear (Ursus maritimus) that developed a sudden onset of seizure-like activity and died. The medium and large cerebral arteries of the meninges had moderate to severe diffuse discoloration and mineralization of the matrix of the tunica media, with little or no associated cellular reaction. Scanning electron microscopy of the affected arteries showed discrete crystalline calcified deposits in the media and sclerosis of the arterial wall. There were no lesions in the brainstem. The findings suggested a sudden and rapidly fatal loss of blood flow to the brain caused by long-standing arterial lesions. Incidental findings included numerous 0.1- to 10-cm-diameter, hepatic cysts lined with hyperplastic biliary epithelium, a unilateral, unipolar, 3-cm-diameter renal tubular adenoma, and approximately 250 active Baylisascaris sp. nematodes in the intestines.     

Aortic-iliac thrombosis in horses

Vascular obstruction of the hindlimbs was diagnosed clinically in 15 horses, and was characterized at necropsy in eight of those horses. The condition was identified as an incidental post mortem finding in two additional horses. The principal clinical signs in affected horses were progressive exercise intolerance and hind-leg lameness. At post mortem the oldest lesions were located at the aortic quadrifurcation and in the distal portions of the femoral and internal iliac arteries, and consisted of partially or completely occlusive masses of well-organized and well-vascularized fibrous tissue, occasionally containing hemorrhagic or degenerate areas. Proximal to these organized masses, large unorganized thrombi were often present. In the region of the occlusive masses, the tunica intima was obliterated, except for the internal elastic lamina which usually remained intact. The tunica media was largely unaffected, except for ischemic necrosis of the media in greatly distended arteries or under thick plaques. The pathogenesis of the lesions is unresolved. The lesions may result from organization of strongyle-related thromboemboli or the inciting cause may be progressive enlargement and organization of spontaneously developing fibrous intimal plaques. Hypercoagulability of the blood may have contributed to thrombosis in one mare with the nephrotic syndrome. Routine examination of the aortic quadrifurcation and its major branches is recommended in order that subclinical changes may be detected and the natural history of the lesion elucidated.