An ultrastructural characterization of the ooplasm in ovarian follicles of the immature ostrich (Struthio camelus)

Primordial, pre-vitellogenic and vitellogenic follicles were present in the ovary of the immature ostrich. The oocytes of these follicles were composed of a nucleus surrounded by ooplasm. Central, intermediate and cortical regions formed the ooplasm. The organelles present in these ooplasmic regions varied depending on the stage of follicular development. In primordial and small pre-vitellogenic (100-150 microm in diameter) follicles the central region of the ooplasm was dominated by an accumulation of organelles, which formed Balbiani's vitelline body. In contrast, the central region in vitellogenic follicles was filled with numerous large yolk spheres, many of which contained lining bodies. Numerous lipid droplets interspersed with mitochondria and small yolk spheres formed the intermediate ooplasmic region in primordial and small pre-vitellogenic follicles. In large pre-vitellogenic (150-400 microm in diameter) and vitellogenic follicles the intermediate region contained a greater density of mitochondria and small yolk spheres. Small yolk spheres were observed in the cortical region of pre-vitellogenic follicles. An interesting feature of the cortical region in vitellogenic follicles was the frequent occurrence of Golgi complexes. The results of the study indicate that although the ovarian follicles in the immature ostrich are not ovulated, the components and composition of the ooplasm are similar to those observed in the mature follicles of other avian species.     

Protothecal granulomatous meningoencephalitis in a dog

A case of central nervous system protothecosis in a dog is reported. A three-year-old male Maremma sheepdog was referred with a two month history of diarrhoea associated with progressive tetraparesis, depression and right circling. Stupor, severe proprioceptive deficits, bilateral decreased thoracic limb flexor reflexes and bilateral deficit of the menace reaction were detected on neurological examination and a multi-focal neurological localisation was suspected. Histopathological evaluation revealed multi-focal granulomatous foci in the thalamus, hippocampus and caudal brainstem containing numerous oval-rounded organisms with a thick, periodic acid-Schiff-positive and Gomori's methenamine silver-positive cell wall, a basophilic cytoplasm and one nucleus. Scattered sporangia containing two to four endospores were also observed. Morphological features were consistent with Prototheca species. Ultrastructurally, numerous degenerated algae were present within macrophages mainly lacking nuclei and cytoplasmic organelles. Generally, protothecosis in dogs is characterised by systemic signs because of a multi-organ involvement, and haemorrhagic colitis or ophthalmologic signs are the most frequent presenting signs. However, protothecosis should be added, also in Europe, to the list of the differential diagnoses in adult dogs with a multi-focal neurological localisation even in absence of other clinical signs.     

Nasal rhinosporidiosis in two dogs

Nasal rhinosporidiosis was diagnosed in 2 dogs. Cytologic criteria for diagnosis were the presence of 5- to 10-microns endospores and 50- to 1,000-microns sporangia. These findings made it easy to differentiate rhinosporidiosis from the more common nasal mycoses such as cryptococcosis. Treatment is principally surgical, but medical management can be performed if the lesion is inoperable or recurs despite multiple surgeries. Dapsone is one drug that has been used in such patients.     

Ultrastructural and cytochemical studies in normal Japanese quail (Coturnix coturnix japonica) eosinophils and in those from birds with experimentally induced eosinophilia

Normal eosinophil development in the Japanese quail (Coturnix coturnix japonica) was similar to that described in the fowl and the duck, with granulogenesis occurring in the Golgi apparatus. The characteristic lipid droplets were small in the immature eosinophils, and after staining specifically for lipid, small moieties were also traced to the Golgi apparatus. In mature eosinophils the lipid droplets measured between 1.0 and 1.5 micron in diameter and they were surrounded by profiles of smooth endoplasmic reticulum. Eosinophilia was difficult to induce in quails; injections of either horse serum or bovine serum albumin (BSA)/aluminium hydroxide produced a poor response. In some quails in which eosinophilia was produced, however, eosinophil granules showed many crescentic and vacuolated forms. The lipid droplets in the activated eosinophils were fused in many cells to form large intracellular aggregates of lipid. Quail eosinophils, which hitherto have been regarded as peroxidase-negative, had strong activity in the lipid droplets of cells from stimulated birds. It is postulated that this peroxidase-positive reaction may represent a form of ceroid or lipofuscin pigment resulting from lipid peroxidation. Acid phosphatase and trimetaphosphatase reactions were reduced in many activated cells, with a large proportion of granules being non-reactive. The results of dietary manipulations in quails appear to suggest that in stressful situations the eosinophil metabolism is altered and there is a reduction in the number of lipid droplets in the cell.     

Cytologic identification of immature endospores in a dog with rhinosporidiosis

An 8-year-old, intact, male Labrador Retriever was presented to the Boren Veterinary Medical Teaching Hospital at Oklahoma State University with a 2-month history of severe sneezing episodes that resulted in epistaxis and bilateral sanguineous discharge. Rhinoscopy revealed a small polypoid mass, and specimens were obtained for histopathology. Microscopic examination of formalin-fixed tissue specimens revealed organisms consistent with Rhinosporidium seeberi. The mass was surgically excised and impression smears were made for cytology examination. Smears revealed high numbers of endospores, typical of those previously described for R seeberi. In addition, numerous smaller structures, presumed to be immature endospores, were noted. The immature endospores were morphologically distinct from mature endospores and have not been described previously. Recognition of immature forms of Rhinosporidium may help prevent misidentification of the organism or misdiagnosis of a dual infection.     

Morphology of Sarcocystis gigantea in experimentally-infected sheep

The development of the parasite and lesions was studied in 32 sheep killed 10 days to 47 months after inoculation with Sarcocystis gigantea sporocysts from cats. At 21-42 days post-inoculation (d.p.i.), there was a mild encephalitis, but organisms were not seen in the brain. Immature sarcocysts were detected from 40-84 d.p.i. The cyst wall was not measurable by light microscopy at 40 d.p.i., but was 1.5-2 microns thick at 84 d.p.i. At 119 d.p.i. both immature cysts containing only metrocytes, and mature cysts containing both metrocytes and merozoites, were present. These mature cysts did not have a secondary cyst wall. A mature cyst, 350 microns in length, was found in a sheep killed at 8 1/2 months p.i. At 10 m.p.i. cysts were up to 0.5 mm long and a secondary cyst wall was present. At 47 m.p.i. cysts were 2-5 X 4.5-7.5 mm, and were found only in the muscles of tongue, oesophagus, pharynx and flank.     

Draining skin lesion from a desert poodle

A 16-month-old intact female Poodle in Arizona had a history of intermittent coughing of a few weeks duration. Coccidiomycosis antibody screening test results were negative for immunoglobulin (Ig) M, but were positive (1:64) for IgG. Fine needle aspiration specimens of a draining lesion on the right palmar front foot contained large numbers of neutrophils, many of which contained bacteria, and lower numbers of macrophages. A few small structures also were observed, 2-5 microm in diameter with thin, nonstaining capsules and small, round to oval densely aggregated, eccentric nuclei. Cytologic findings were consistent with septic pyogranulomatous inflammation with Coccidiodes immitis endospores. Fungal culture of a sample from the draining lesion yielded white growth with barrel-shaped arthroconidia. Identification of the organism as C immitis was confirmed by a commercial DNA probe test. Although coccidioidomycosis often is diagnosed by microscopic identification of C immitis spherules in cytologic specimens, in this case only endospores, which are released from mature spherules, were observed. In cases of suspected coccidiodomycosis, the unique morphology of endospores may be useful in making a cytologic diagnosis.     

ULTRASONOGRAPHIC APPEARANCE OF CANINE GASTRIC POLYPS

Benign gastric polyps are uncommon in dogs and most are discovered incidentally. Polyps protruding into the pyloric antrum can cause gastric outflow obstruction. Clinical and ultrasonographic findings in seven dogs with histologically confirmed benign mucosal gastric polyps were reviewed. Sonographic findings such as shape, size, echogenicity, location, evidence of gastric wall thickening, wall layering, and size of regional lymph nodes were recorded. Five sessile and two pedunculated masses of different sizes (range 7–60 mm) and echogenicities were found. They primarily arose from the mucosal layer and protruded into the gastric lumen. Only one dog had a large inhomogeneous mass with a poorly visualized gastric wall layering. The polyps were all single, and were located in the pyloric antrum in six out of seven dogs. Although the ultrasonographic appearance allowed a presumptive diagnosis of mucosal gastric polyp, the final diagnosis was determined from histopathologic examination.     

延边黄牛卵母细胞与体细胞克隆胚胎内脂滴变化规律的研究

用苏丹Ⅳ和苏丹黑B 2种染色方法,对体外培养的延边黄牛各发育阶段卵母细胞和体细胞克隆胚胎内脂滴变化进行研究,并对2种染色方法进行了比较。结果表明,随着细胞发育阶段的不同,脂滴的含量也随之变化,从未经成熟培养的卵母细胞到8细胞期胚胎内脂滴不断增多,脂滴直径不断增大;而从8细胞期到囊胚期胚胎内脂滴不断减少,脂滴直径也不断减小。也显示苏丹IV染色效果较好,可作为测定活体细胞中脂滴的检测方法。     

Proliferation of canine intervertebral disk chondrocytes in three-dimensional alginate microsphere culture

Proliferation of chondrocytes from nucleus pulposus (NP) and anulus fibrosus (AF) was confirmed in three-dimensional culture using alginate microspheres. Cells isolated from NP and AF were incorporated in microspheres and cultured for 14 days. Round mononuclear cells of 20-25 microm in diameter proliferated and formed aggregates. At day 14, alcian blue positive matrix surrounded the proliferating cells. The cells had cytoplasmic vacuoles stained positively by toluidine blue. On electron microscopy, the cells contained proteoglycan vacuoles and lipid droplets in the cytoplasm and synthesized collagen fibrils and electron dense granules surrounding the cell. These features of the cells were characteristic for chondrocytes. This culture system should be useful to further investigate metabolic activities of intervertebral disk chondrocytes.     

Evaluation of the Lipid Content in Bovine Oocytes and Embryos with Nile Red: a Practical Approach

In this study, the fluorescent lipid dye Nile Red, was used to demonstrate that the lipid content of immature bovine oocytes is correlated with the morphological appearance of the ooplasm. Oocytes with a uniform dark cytoplasm contained significantly more intracellular lipids in lipid droplets compared with oocytes with a granulated or pale cytoplasm (p < 0.05). Furthermore, this lipid-analysing technique was applied for the first time on single bovine in vitro embryos, showing a significant increase of the lipid content in lipid droplets after culture in the presence of serum (p < 0.05).     

Ultrastructure of Sarcocystis spp. from donkeys (Equus asinus) in Egypt

The fine structure of Sarcocystis spp. from donkeys (Equus asinus) in Egypt is described. Sarcocysts were found in the oesophagus, diaphragm and heart of 18 of 20 donkeys. Only one type of mature muscle cyst was found. Sarcocysts were 120-410.6 X 48.4-50.2 microns. The primary cyst wall had numerous 3.3-3.7 microns villi. Each villus contained 20-60 fibrillar elements which extended from the ends and sides of the villi throughout the ground substance, where they became tightly packed. The bundles of fibrillar elements formed junctions with the pellicles of the metrocytes. Ultrastructurally, Sarcocystis spp. of the donkey was similar to sarcocysts previously described from the horse.     

Comparative studies on lipid analysis and ultrastructure in porcine and southern minke whale (Balaenoptera bonaerensis) oocytes

The present study was conducted to clarify the difference in the color of the cytoplasm in immature follicular oocytes from prepubertal and adult minke whales. The four lipid contents (triglyceride, total cholesterol, phospholipids and non-esterified fatty acids) in vitrified immature oocytes from prepubertal and adult minke whales, and also in fresh and vitrified immature porcine oocytes, were measured. The lipid contents in vitrified-warmed minke whale oocytes were similarly high compared with those in vitrified-warmed porcine oocytes. In particular, the total cholesterol and phospholipid contents in the vitrified immature oocytes from prepubertal and adult minke whales were significantly (P<0.05) higher than those from prepubertal pigs. Furthermore, the distribution of lipid droplets in fresh and vitrified immature oocytes was observed in transmission electron microscopy. Lipid droplets in the prepubertal minke whale oocytes were distributed throughout the cytoplasm. In contrast, adult minke whales had larger lipid droplets which were distributed mainly in the central portion of the cytoplasm. The lipid droplets of immature oocytes from prepubertal pigs were larger than those in minke whale oocytes. These results indicated that the difference in the distribution of the cytoplasmic lipid droplets may result in the difference in the color tone of both prepubertal and adult whale oocyte cytoplasm.     

Locations and types of neoplasms in immature dogs: 69 cases (1964-1989).

Sites, histologic types, and frequencies of neoplasms in immature dogs (less than or equal to 6 months old) were evaluated from data collected over 25 years. The frequencies of neoplasms in immature dogs were compared with those of mature dogs (greater than 6 months old). Of 69 immature dogs with neoplasms, 5 had 2 primary neoplasms each, resulting in a total of 74 neoplasms. The 3 most common sites for neoplasia, in decreasing order, were the hematopoietic system, brain, and skin. Immature dogs were 10.9 times more likely to have a neoplasm located in the brain, compared with mature dogs. Immature dogs also were 3.3 times more likely to have a neoplasm associated with the hematopoietic system, compared with mature dogs.     

Morphologic and morphometric study of the facial nerve in clinically normal adult dogs.

A morphologic and morphometric study was carried out on the facial nerve: to determine normal histologic data in myelinated fibers of clinically normal young adult dogs; to establish reference values for mean fiber diameter, and to delineate the relative diameter frequency distribution curve. Few degenerative changes were observed in single teased-nerve fibers and semithin cross-sectional nerve preparations. Statistical difference was not observed between left and right facial nerves. The distribution of fiber diameters in the facial nerve was unimodal. Mean (+/- SD) fiber diameter of the facial nerve was 3.92 +/- 1.18 microns. Approximately 89% of fibers in the facial nerve had diameter between 3 and 6 microns. Fiber diameter ranged from 2 to 12 microns; however, less than 1% of fibers had diameter greater than 8 microns.     

More than meets the eye: subretinal aspirate from an acutely blind dog

A 14-year-old, spayed female Cocker Spaniel was presented to the Boren Veterinary Medical Teaching Hospital at Oklahoma State University with acute loss of vision in the right eye and a history of intermittent bloody diarrhea of unknown duration. Small, white, plaque-like lesions in the retina and subretina were visualized by direct ophthalmic examination, and aspirated with ultrasound-guidance. A direct smear of the subretinal fluid was highly cellular and contained large numbers of pleomorphic organisms consistent with Prototheca sp. The structures were round, oval, or elongated, 4-6 microm width and 8-16 microm in length and surrounded by a thin, clear cell wall. Small, central, pink to purple nuclei were observed in some organisms, but in most, the nuclei were obscured by a deeply basophilic, granular cytoplasm. Some organisms contained endospores. Negatively-stained structures of similar size and shape were considered to be empty casings (theca) of ruptured sporulating and nonsporulating forms of the organism. Protothecosis usually is a disseminated, fatal disease in dogs. The Prototheca organisms observed in this case showed characteristic morphology, illustrating the ability to diagnose protothecosis in cytologic samples.     

Development of Babesia gibsoni in the midgut of larval tick, Rhipicephalus sanguineus.

Studies were made on the development of Babesia gibsoni in the midgut of the larval tick, Rhipicephalus sanguineus. Six hr after repletion, merozoites of B. gibsoni, freed from erythrocytes, were observed in the midgut contents of the tick. After that, within 24 hr, those merozoites were transformed into the ring-forms which were relatively large, 2-3 microns in diameter. Later, the ring forms developed into the spherical forms which were subelliptical in shape and 4-6 microns in diameter. Within 2-4 days, the elongated forms, 5-8 microns in length, were found. At this time, some of the binucleated fusion form has assumed a form intermediate between the spherical and elongated-forms. About 5-6 days after repletion, large round or elliptic zygotes, 8-10 microns in diameter, were observed in the tick gut.     

Ultrastructural features of atretic follicles in the sexually immature ostrich (Struthio camelus)

The aim of this study was to describe the ultrastructural features of atresia in follicles of the immature ostrich (12–14 months old); a ratite that displays seasonal, precocious ovarian activity. The early stage of atresia in primordial, pre-vitellogenic and vitellogenic follicles was characterized by the accumulation of lipid droplets in the granulosa cells. Granulosa cells with condensed cytoplasm and nuclei were a prominent feature during the intermediate phase of atresia. The degenerating follicles were then infiltrated by stroma during the terminal stages of atresia. The results of this study provide further information on the morphology of atretic ovarian follicles in the immature ostrich.     

Malignant hyperthermia susceptibility: cardiac histomorphometry of dogs and young and market-weight swine.

The defect causing malignant hyperthermia has been proposed to involve cardiac as well as skeletal muscle. We tested the hypothesis that histomorphometric parameters for ventricular wall from malignant hyperthermia-susceptible swine and dogs were abnormal. Hearts were obtained from: mature dogs, age- and weight-matched young swine (89 +/- 15 days, 30 +/- 3 kg); and market-weight swine (102 +/- 10 kg). Using light microscopy, estimates were made for muscle nuclear dimensions and the volume-fraction of nuclei, sarcoplasm, blood vessels, and interstitial space. Cardiac maturation in both MH and normal swine was accompanied by decreased myocyte volume-fraction due to decreased nuclear volume-fraction and increased interstitial space volume-fraction. Sarcoplasm and vasculature volume-fraction were unchanged after maturation. Nuclear volume-fraction was slightly greater (p less than 0.05) in the right ventricle than the left for malignant hyperthermia and normal swine. Myocyte nuclear dimensions were generally similar among animals. Dogs and the oldest group of swine were not significantly different. Myocytes of all swine contained multiple nuclei, closely spaced in rows of 2 to 12. In contrast, most myocytes of mature dogs apparently contained one or two nuclei. Histomorphometric values were not significantly different between normal and malignant hyperthermia young swine and dogs. However, within the market-weight swine, volume-fraction for malignant hyperthermia myocytes and myocyte nuclei was decreased and interstitial space was increased compared to normal.(ABSTRACT TRUNCATED AT 250 WORDS)     

Distribution of angiostrongylosis in Cornwall

One hundred and ninety-seven faecal samples were examined for Angiostrongylus vasorum larvae, from dogs throughout Cornwall, which were presented with one or more clinical signs consistent with infection, were known to eat slugs or snails or were related to an infected dog. Twenty samples (10-1 per cent) contained nematode larvae, of these A vasorum larvae were identified in eight (4-1 per cent) and larvae of Filaroides species in eight (4-1 per cent). Seven of the eight dogs positive for A vasorum were found to have been infected within an area six miles in diameter incorporating Redruth. Statistical analysis suggested that this aggregation of cases was unlikely to be a random event (P < 0–01). No correlation between published data on the distribution of the intermediate host, slugs and snails and the cases of A vasorum infection reported in this paper could be found. It is concluded that A vasorum infection is enzootic within a small area around Redruth in the mid-west of Cornwall.