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.     

猪旋毛虫病膈肌的病理形态学观察

对80份猪旋毛虫病膈肌进行了病理组织学和组织化学观察,并对其中5份作了透射电镜观察。结果:膈肌旋毛虫病灶的病变分为非包囊型、包囊型、包囊性肉芽肿型、肉芽肿型和淋巴细胞结节型等5种类型;旋毛虫包囊壁内、外两层的成分是胶原,内层来源于感染肌纤维,外层为增厚的感染肌纤维的基底膜;感染和非感染肌纤维,均具有变性和再生变化,但前者的再生最后形成包囊,而后者则一般以结缔组织取代之;感染和非感染肌纤维均有一定的碱性磷酸酶活性,尤以肌膜最明显,而非特异性酯酶仅位于旋毛虫病灶内;旋毛虫病灶内,T、B淋巴细胞同时存在,且尚见嗜酸性白细胞。     

Invasive teat surgery in dairy cattle: part I - surgical procedures and classification of lesions

A prospective study was performed to identify the nature and management of teat abnormalities in cows presented to a referral teaching hospital during a three year period. All cattle (n = 60) admitted to the Ontario Veterinary College for teat problems were evaluated by physical examination; in 53 teats, contrast radiography or xeroradiography were obtained. Surgery was performed on 52 teats from 51 cows and a prosthesis was implanted in 27 teats. Short term (under two weeks) complications included intraoperative bleeding (n = 6), milk leakage through the incision (n = 4), and failure to milk by machine in 26 cases. Histopathological diagnosis of sections taken from obstructive lesions included fibrous tissue (n = 8), normal mammary tissue (n = 3), fibropapilloma, mammary polyps, and inflamed mucosa (one each). The lesion could be classified into five types: 1) focal teat cistern obstruction, 2) diffuse teat cistern obstruction, 3) membranous obstruction, 4) diffuse teat and gland cistern obstruction, or 5) leakage of milk through an abnormal route (i.e. teat fistula, webbed teat, or lacerations).     

Developmental states of the collagen content, distribution and architecture in the pectoralis, iliotibialis lateralis and puboischiofemoralis muscles of male Red Cornish x New Hampshire and normal broilers

1. Developmental states of the collagen content, distribution and architecture in the pectoralis (PT), iliotibialis lateralis (ITL) and puboischiofemoralis (PIF) muscles of male Red Cornish x New Hampshire (RN, 80 d, body weight 2.9 kg) and normal (3.1 kg) broilers were evaluated. 2. In PT muscle the total amount of collagen was significantly greater in RN broilers (3.33 mg/g) than in normal ones (1.71 mg/g). This higher collagen content in RN broilers was based mainly on the closer mesh sizes of endomysial honeycomb. The collagen structures in the perimysia also differed between broiler types, when more collagen fibres were observed in RN broilers. 3. ITL muscle contained total collagen of 4.10 to 5.00 mg/g. Types I and III collagens were distributed on the perimysia at higher percentages in RN broilers (31.6%, 37.2%) than normal (15.6%, 30.8%), respectively. The thick bands of tough collagen fibres characteristic of ITL muscle perimysium in cockerels had not yet developed in these broilers. 4. Total collagen was 4.63 to 6.29 mg/g in PIF material with fascia. In PIF muscle the perimysial collagen fibres had not yet attained their full growth but consisted of densely packed fibrils. PIF muscle was characterised by the earlier maturing collagen structure. 5. These results show that a perimysial collagen structure in broilers is still in an undeveloped state. It is supposed that tenderness of broiler meat is attributed mainly to characteristics of the collagen distribution, in which the majority of types I and III collagens is distributed on the closer mesh of endomysial honeycomb.     

Topographic Distribution of the Different Cell Types,Connective Tissue and Vascular Tissue/Lumina Within a Functional Bovine Corpus Luteum and its Association with Breed,Type of Fixation Protocol and Stage During the Cycle

In the present study, we analysed the effect of fixative, breed, luteal stage and location on the nuclear density, volume density of connective tissue and vascular tissue/lumina within a bovine luteal gland in view of the development of an in vivo sampling technique to longitudinally monitor luteal histophysiology. The inner zone defined as the zone geometrically closest to the centre of the gland shows a significantly lower nuclear density (for all cell types) and a higher volume density of collagen fibres and vessels when compared with the outer zone (p < 0.001). The nuclear density in luteal glands from Holstein‐Friesian cows is not significantly different from that in Belgian Blue cows, nor is it in stage II vs stage III glands. The collagen fibre content was significantly lower in glands of Belgian Blue cows (p = 0.01) and in younger glands (p = 0.003). Hence, it seems that the lower nuclear density in the inner zone was compensated by a higher amount of collagen fibres. As the type of fixative applied has a significant effect on the nuclear density of the different cell types, the present study warrants future research to further optimize the fixation protocol. As a conclusion, we can state that the topographic difference in nuclear distribution for the different cell types in a bovine luteal gland is only significant when comparing the inner vs the outer zone. This implies that if a sample representative for the whole gland has to be taken, for example, when taking an in vivo sample, it is necessary that the biopsy goes through the inner zone and contains the total diameter of the gland.     

Collagenous and elastic system fibres in the aorta of cattle poisoned by Solanum glaucophyllum

The proportions of fibres of the collagenous and elastic systems were measured in the aortas of three normal heifers and in nine heifers given the calcinogenic plant Solanum glaucophyllum for 15, 30 or 60 days. There were decreases in the amount of collagen relative to reticulin, and in the proportion of elastic fibres which were related to the period of dosing. These changes may have an adverse influence on the animals' cardiovascular function.     

Histology and ultrastructure of the developing superficial digital flexor tendon in rabbits

This study was designed to investigate the developmental changes in the superficial digital flexor tendon (SDFT) of the white New Zealand rabbit, from 5 to 7 days pre-natal to 112 days post-natal (PN), at histological and ultrastructural levels. The tendons changed from being highly cellular with 13079 ± 2538 cell/mm² and little directional organization to a longitudinally oriented, predominantly connective tissue structure with relatively few, mature tenocytes (2384 ± 365 cell/mm²). Fibrillogenesis was seen in isolated vacuoles of fibroblast cytoplasm during fetal life as well as after birth, but less frequently with increasing age. At the ultrastructural level, there was a progressive increase in the mean diameter of collagen fibrils with age throughout the population , until PN day 28. A bimodal distribution of collagen fibril size was first observed on PN day 56 while at day 112, the fibrils were fully differentiated and showed a multimodal size distribution. The development of elastic fibres preceded that of collagen fibres and accompanied progressively more marked sinusoidal crimping in collagen fibres on PN days 7 and 14. The tendons of older animals became less tightly crimped. The cells of the epi- and endotenon were less mature and relatively undifferentiated compared with the cells in the body of the tendon of the same age, which might explain their ability to initiate healing of tendon injuries in older animals. In conclusion, extensive changes were observed in tendons during growth and maturation, with diameters of collagen fibrils, tissue orientation and cellularity being the parameters affected, probably due largely to increased physical activity.     

Elastic Fibre System of the Female Canine Urethra. Histochemical Identification of Elastic, Elaunin and Oxytalan Fibres

The elastic system fibres of the female urethra were investigated in seven dogs of different breeds. The fibres were stained and differentiated with orcein, Verhoeff's iron hematoxylin, and pararosanilin-based aldehyde fuchsin, with and without previous oxidation. Orcein and aldehyde fuchsin revealed some subepithelial longitudinally orientated delicate fibres (elaunin fibres) and numerous coarse longitudinal fibres (elastic fibres) in the deeper subepithelial connective tissue containing the vascular plexus, as well as a network of fibres of different calibers in the periurethral connective tissue. Elastic system fibres were also found in association with the sinusoids of the vascular plexus and the urethral smooth musculature. Verhoeff's iron hematoxylin only reacted distinctly with coarse (elastic) fibres. When applied following oxidation, aldehyde fuchsin disclosed an extensive meshwork of additional delicate fibres (oxytalan fibres) in the subepithelial connective tissue adjacent to the basement membrane. Oxytalan fibres were also discernible in the sinusoidal adventitia of the vascular plexus, as well as between smooth and striated muscle fibres. Due to the predominantly longitudinal orientation of the elastic system fibres and the low urethral resistance to manual expression of urine from the bladder post mortally, when the elasticity of the urethra is still intact and the activity of other continence factors can be excluded, the elastic tissue is not judged to be a major contributory factor to urinary continence.     

Hereditary equine regional dermal asthenia ("hyperelastosis cutis") in 50 horses: clinical, histological, immunohistological and ultrastructural findings

Data on fifty horses with hereditary equine regional dermal asthenia (HERDA; "hyperelastosis cutis") were collected on clinical, histopathological, ultrastructural and immunohistological findings. All horses were Quarter horses or of Quarter horse ancestry. Pedigree evaluation strongly supported an autosomal recessive mode of inheritance. The most common lesions were seromas/haematomas, open wounds, sloughing skin, and loose, easily tented skin that did not return to its initial position. Definitive diagnosis could not be made via histopathology, although the presence of tightly grouped thin and shortened collagen fibres arranged in clusters in the deep dermis was suggestive of the disease. Trichrome, acid orcein-Giemsa and immunohistochemical stains for collagens I and III showed no consistent abnormalities compared to control horses; an increase in elastic fibres was not a consistent finding. Electron microscopy showed no abnormalities in the periodicity of the collagen bundles; neither orientation nor variation of cross-section diameter of the collagen fibrils differentiated control from affected horses. The diagnosis of HERDA relies on clinical presentation, but may be supported by suggestive (although not pathognomonic) histopathological lesions.     

Autofluorescence of bovine ligamentum nuchae, cartilage, heart valve and lung measured by microscopy and fibre optics

The fluorescence of bovine tissues was measured post mortem by microscopy of frozen sections and by using optical fibres to excite fluorescence and to measure fluorescence emission spectra. Mechanical disruption of the tissue (by comminution or sectioning) did not appreciably change tissue fluorescence spectra. Ligamentum nuchae had the strongest fluorescence and lung tissue had the weakest. In samples measured with a minimum prior exposure to ultraviolet light, the peak fluorescence emission was at 410 or 420 nm (with excitation at 365 nm). Exposure to ultraviolet light for about 1 minute shifted the fluorescence peak to 450 to 470 nm. Further exposure (about 30 minutes) caused a loss of the 450 to 470 nm fluorescence peak, while emissions above 530 nm were maintained or strengthened. Microscopy showed that the fluorescence that was measured by fibre optics from intact connective tissues originated mostly from collagen and elastin fibres.     

Horse lumbrical muscle: possible structural and functional reorganization in regressive muscle

An anatomical study of horse lumbrical muscle (Lm) was carried out by light and electron microscopy in combination with immunochemical and cytochemical methods. Paraffin sections were subjected to haematoxylin and eosin (H & E) and Masson's trichrome staining for morphometric analysis. Paraffin sections were also used for immunostaining by anti-PGP 9.5 for reaction with nerve-protein associated-structures, anti-heat-shock protein 70 (hsp 70) for detection of gene expression changes, anti-fast myosin for the determination of muscle fibre types, and for detection of apoptotic gene expression of muscle fibres by the TUNEL method. The distribution of muscle fibre types on frozen sections was also examined by assaying ATPase (pH 4.2). We found that the proximal end of the tendon of the unipennate-shaped Lm binds to the deep digital flexor tendon, and the distal end of the Lm tendon connects to the medial surface of the palmar annular ligament. The Lm was not always present, but when found it varied in length greatly, up to 8 cm (muscle part alone), and weighed less than 1 g. The Lm was white, pale, or reddish in colour depending on the ratio of muscle fibre and connective tissue contents. The semi-tendinized regressive Lm was composed of rich vasculature, peripheral nerves, and nerve-like organs similar to the neuromuscular spindle (NMS). The extrafusal muscle fibres (e-lm) that surround the NMS were replaced with a thick outer capsule of connective tissues (CT) in the Lm nerve-like organ, which we named the neurotendinous capsule (NTC) organ. NTC organs exist alone or as multiple structures (up to eight) surrounded by a common outer capsule at the outermost CT ring. The NTC possesses several intrafusal muscle fibres (ifm) just as the NMS does. That the ifm was associated with nerve endings was confirmed by anti-PGP 9.5 and electron microscopic observation. Some muscle fibres in ifm and e-lm reacted with anti-fast twitch myosin and with anti-hsp 70. The e-lm exhibited at least two fibre types, determined by ATPase (pH 4.2) assay. The ifm exhibited mainly type I (slow twitch) fibres. No apoptotic gene expression was detected in either ifm or e-lm, suggesting the Lm is a vital organ. The degenerating fibres observed in ifm and e-lm indicate that the turnover rate of cytoplasmic components is accelerated. We attribute this phenomenon to the necessity for adaptation to new environmental demands. The surprising finding of tubular aggregates (TAs) in ifm of the NTC organ suggests that the Lm is continuously adapting. Some results related to variation in diameter of the collagen fibrils, isolation of the NTC organ and the myofibrillar protein constituents are also discussed. In conclusion, the so-called regressive Lm has rich vasculature, many peripheral nerves, and newly described NTC organs. The induction of heat-shock protein, lack of apoptotic gene expression in ifm and e-lm fibres, and TA formation in ifm suggest that horse Lm responds to environmental stress through reorganization and/or remodelling of cell constituents. We hypothesize that the horse Lm has lost its original role as a contractile element and changed to another function, likely as a vital nerve organ.     

Development of collagen fibres and lysyl oxidase expression in the presumptive dermis of chick limb bud

Lysyl oxidase (LOX) plays a critical role in the formation of cross-linkages in extracellular matrix molecules. Thus, it is essential for the biogenesis and homeostasis of the connective tissue matrix. During development, collagen fibres and elastic system fibres emerge and accumulate in a temporospatial manner in the presumptive dermis of chicks. In this study, we investigated LOX mRNA expression by laser capture microdissection and RT-qPCR and LOX protein localization by immunohistochemistry. The picrosirius polarization method was used to investigate a relation between collagen accumulation and LOX expression. PCR analysis showed that the expression of LOX mRNA in the presumptive dermis became apparent at embryonic day 13 and increased considerably by ED17. Immunohistochemical staining for LOX in the dermis was very low at all stages of development. Accumulation of collagen fibres was seen in the dermis on ED10, and higher wavelengths of birefringence became evident by ED13. Our findings suggest that the temporal pattern of LOX mRNA expression correlates with collagen fibre accumulation in the dermis of the developing chick limb bud, whereas LOX expression was relatively constant at the protein level.     

Anatomy of the Sinus Node of Camels (Camelus dromedarius)

Anatomy of the sinus node was studied in six camel hearts (Camelus dromedarius) with serial histologic sectioning. The sinus node in this species of animal was located 0.5 mm beneath the epicardium, near the junction between the cranial vena cava and the right atrium at the sulcus terminalis. Its shape was elongated, bent oblong with 28.25 mm length, 5.75 mm width and 5.38 mm thickness. The maximum section area was 101.66 mm². The central artery of the node originated from the circumflexus branch of the left coronary artery and, throughout its length in the substance of the sinus node, had an internai elastic membrane. Histologically, the sinus node of this animal contained a central artery and a framework of collagen fibres, which were distributed around the central artery. The nodal cells were irregularly organized around the central artery and two types, i.e.‘p’ cells and transitional cells were present. The ‘p’ cells had a perinuclear clear zone but the transitional cells contained more myofibrils. The intercalated dises were not present. At the periphery of the sinus node there were many nerve fibres and a ganglion. The purkinje fibres were present within atrial myocardium, as well as within ventricular myocardium. The glycogen content of the sinus nodal cells was higher than that of the atrial myocardial cells.     

Hereditary equine regional dermal asthenia in three related Quarter horses in Brazil

Hereditary equine regional dermal asthenia belongs to a group of inherited, congenital connective tissue dysplasias usually described as hyperelastosis cutis, cutaneous asthenia, dermatosparaxis, or Ehlers-Danlos-like syndrome. This report presents the clinical and histological features of three related Quarter horses affected with regional dermal asthenia. These horses had bilateral asymmetric lesions of the trunk and lumbar regions, where the skin was hyperextensible. Handling of the skin elicited a painful response and superficial trauma led to skin wounds. The skin was thinner than normal in the affected areas, with thickened borders and harder fibrotic masses (pseudotumours). The histopathological findings included thinner and smaller collagen fibrils, and a loose arrangement of collagen fibres in the middle, adventitial and deep dermis. Masson's trichrome and Calleja stains did not reveal any abnormality of collagen and elastic fibres. Electron microscopy showed no abnormalities. As in human patients, pseudotumour histopathological findings included fibroplasia and neovascularization. The pedigree chart of these animals supports an autosomal recessive type of inheritance, which has been suggested by other studies. This is the first report of this disease in Brazil. Its clinical and histological features resemble those described in horses affected with this condition in the United States.     

Immunocytochemical localisation of carbonic anhydrase isozyme III in equine skeletal muscle

The location of carbonic anhydrase III (CA-III) in frozen sections of biopsies of Thoroughbred horse skeletal muscle was studied. Fibre types were determined by ATP-ase and succinate dehydrogenase staining. CA-III isozyme was detected using a peroxidase conjugated anti-CA-III antibody. CA-III was found to be localised in slow twitch oxidative fibres (ST), but was also present in fast twitch oxidative (FTH) fibres in small amounts. Fast twitch glycolytic (FT) fibres were stained lightly compared with control sections. The concentrations of CA-III in muscle and liver were 70 micrograms/mg protein and 4 micrograms/mg protein, respectively. CA-I and CA-II were not found in muscle extracts by the double immunodiffusion method.     

Type,diameter and distribution of fibres in some respiratory and abdominal muscles of the goat

This study was designed to determine the histochemical properties, size and composition of fibres in the diaphragm, intercostal and abdominal muscles of goats to clarify whether reported similarities in respiratory muscle physiology between goats and humans have a structural basis. Serial sections (10 µm) of muscular tissue from adult female goats were stained for myosin adenosine triphosphatase and reduced nicotinamide adenine dinucleotide dehydrogenase-tetrazolium reductase activities; the fibres were classified into types I, IIA and IIB; and their mean diameter and composition were determined. Abdominal and intercostal muscles contained types I, IIA and IIB fibres in the ratio 1:1:1, and the mean diameter of the fibres ranged from 49.2 to 62.2 µm. In contrast, the diaphragm contained 58.9% type I and 41.1% type II fibres, and the latter could not be differentiated into types IIA and IIB. Diaphragmatic fibres were also smaller (36.9 – 40.9 µm). These findings contrast with those in humans, where the diaphragm, intercostal and abdominal muscles contain >50% type I fibres and have fibres of identical diameter. The differences in fibre characteristics between the diaphragm, intercostal and abdominal muscles of goats and the differences between goats and humans need to be taken into consideration in interpreting the results from studies in respiratory muscle physiology.Abbreviations mATPase myosin adenosine triphosphatase - NADH-TR nicotinamide adenine dinucleotide dehydrogenase-tetrazolium reductase     

Growth changes of the collagen content and architecture in the pectoralis and iliotibialis lateralis muscles of cockerels

1. Growth changes of the collagen content and architecture in the pectoralis (PT) and iliotibialis lateralis (ITL) muscles were examined using cockerels from 1 to 14 weeks of age. 2. Total collagen content in PT muscle showed little change, but in ITL muscle reached a maximum at 5 weeks and thereafter decreased slightly until 14 weeks. The collagen content was markedly larger in ITL muscle after 5 weeks. Pyridinoline content of collagen increased abruptly from 5 to 14 weeks in both muscles, but no difference between muscle types was detected. 3. The cell size of the endomysial honeycombs increased with the development of myofibres, and the mesh size of the perimysium around the honeycombs enlarged. 4. In both muscles endomysia were an incomplete network of collagen fibrils with many foramina at one week, became a very thin membrane of felt-like fabric in 2 to 5 weeks and thereafter increased in thickness until 11 to 14 weeks. 5. Perimysial width around the secondary fasciculus differed between the muscle types after 5 weeks. In the wider perimysium of ITL muscle, the collagen fibres increased in number and size to make a stack of collagen bands around the fasciculus. In the narrower perimysium of PT muscle, a few platelets of collagen fibres also developed. 6. The perimysial collagen fibre at 1 to 2 weeks had a smooth surface and appeared to be composed of fine collagen fibrils. The fibre at 11 to 14 weeks showed a rugged surface and was composed of coarser collagen bundles that combined with each other into a net-like configuration with very slim meshes. 7. Our results showed that the collagenous components of chicken intramuscular connective tissue changed markedly during the early period of muscle growth in distribution, architecture and quality but with little difference in quantity.     

Milk-flow, ultrasonographic, theloscopic, and histopathological characteristics of the teat in cows with toxic mastitis

The milk-flow, imaging, and histopathological characteristics of the teat in five cows with toxic mastitis were investigated. Teats were grouped into type I (without gross and histopathological abnormality), type II (no abnormal gross findings, but with histopathological abnormality), and type III (with gross and histopathological abnormality). Normal teats from six cows served as controls. Type II and III teats had lower milk-flow, compared with the controls. Ultrasonography revealed no abnormal findings, except for irregular mucosal surface in type II teats and absence of hyperechoic line along the teat canal in type III teats. The theloscopic findings varied from normal to mucosal abnormalities in type II and III teats. Histopathology demonstrated epithelial changes, congestion, hemorrhage, edema, cellular infiltration, and elastic fiber degradation in type II and III teats, with the type III teats showing severe changes. Toxic mastitis was characterized by impaired milk-flow and various degrees of imaging and histopathological abnormality.     

Ultrastructural study of cutaneous lesions in feline eosinophilic granuloma complex

The purpose of this study was to investigate the ultrastructural appearance of flame figures, reported to comprise a mixture of degenerate collagen and degranulated eosinophils, in feline eosinophilic granuloma complex (EGC). Skin specimens from eight cats with EGC and from two clinically healthy cats were examined by transmission electron microscopy. Flame figures appeared to comprise ultrastructurally normal collagen fibrils separated by oedema and surrounded by large numbers of degranulating eosinophils. Longitudinal sections of collagen fibrils displayed the characteristic cross-striation of normal dermal collagen. Feline eosinophils, analogous to human eosinophils, degranulated both by cytolysis and piecemeal degranulation. The results of this study suggest that flame figures form in feline EGC due to eosinophil recruitment and degranulation, and that collagen fibres are partially disrupted but collagen fibrils are not damaged. These findings suggest that eosinophil accumulation and the release of granule contents represent the primary events in feline EGC.     

Staining abnormalities of dermal collagen in eosinophil- or neutrophil-rich inflammatory dermatoses of horses and cats as demonstrated with Masson's trichrome stain

A retrospective study was performed on skin biopsy specimens from horses and cats having eosinophilic granulomas with 'collagen degeneration', eosinophilic skin diseases without 'collagen degeneration' and pyogranulomas without 'collagen degeneration'. Neither the appearance of collagen fibres nor the dominant polymorphonuclear cells, as seen in H & E-stained sections, were predictive of Masson's trichrome findings. Hence, a Masson's trichrome staining abnormality of collagen fibres was no more likely to be present in lesions with 'collagen degeneration' than in those lesions without 'collagen degeneration'. In addition, evaluation of trichrome-stained specimens suggested that there is no collagen degeneration in equine and feline eosinophilic granulomas and that the areas previously referred to as collagen degeneration are more appropriately called flame figures.