Scanning electron microscopic study of bovine leukemic cells

Scanning electron microscopic observation of (i) leukemic cells in peripheral blood and tumor tissues of 11 cattle with bovine leukosis (adult form, n = 5; calf form, n = 2; and thymic form, n = 4) and (ii) peripheral lymphocytes from 2 cattle with persistent lymphocytosis and from 3 healthy cattle revealed morphologic differences of cell surface structures among various forms of bovine leukosis. These differences indicated an interrelationship of cell surface morphologic features between peripheral lymphocytes and tumor cells. Leukemic cells from cattle with the thymic and calf forms characteristically had a smooth surface. In cells from cattle with the adult form, the majority of abnormal cells in the peripheral blood, possessed numerous elongated microvilli on the cell surface, whereas tumor cells in the tissue were pleomorphic with a long villous, plicated surface or had stubby projections. Most of the peripheral lymphocytes from cattle with persistent lymphocytosis were characterized by a dense arrangement of elongated microvilli on the cell surface.     

Prenatal differentiation of bovine oviductal epithelium: an electron microscopic study

In this study, we investigated the ultrastructural changes during the prenatal differentiation of oviductal epithelium in 16 bovine embryos and fetuses from CRL of 18.0 cm to a CRL of 94.0 cm. Ciliated and secretory cells of bovine uterine tube, a derivative of the Müllerian duct, differentiate to distinct development stages in the prenatal period. The typical cellular pattern, which is generally characteristic for the adult bovine oviduct, is also obtained during fetal life. In the early stages (CRL 18.0/20.4 cm), the bovine oviductal epithelium appears mostly undifferentiated. The epithelial cells show only a few mitochondria, some cisternae of rough endoplasmic reticulum (rER) and a small Golgi-complex. Most of the cytoplasm is filled with a large amount of glycogen, which decreases during later development. Interspersed between the undifferentiated epithelial cells, a few cells undergoing ciliogenesis can be observed. Ciliogenesis increased significantly during the later prenatal developmental stages. At a CRL of 55.0 cm, ciliated cells appear fully differentiated with mature cells covering their luminal surface. Formation of cilia usually use the acentriolar pathway. Fibrous granules occurred initially in association with the Golgi-apparatus and r(ER) in the supranuclear cytoplasm. Fibrous granules later fuse with deuterosomes and give rise to procentrioles, which are translocated to the luminal plasma membrane. There they become arranged in a line just beneath the apical cell membrane and further differentiate to basal bodies from which the formation of cilia and striated rootlets take place. Clear signs of differentiation of secretory cells were first seen in our material in fetuses with a CRL of 51.0 cm and 64.0 cm. These cells contain a well developed rER and Golgi-apparatus with dilated cisterns. In the supranuclear cytoplasm, the number of secretory granules continuously increases during later development and the cells adapt to the morphology of mature secretory cells at the CRL 94.0 cm.     

Patterns of bovine T cell-mediated immune responses to bovine herpesvirus 1

Lymphocyte proliferation was used to evaluate T cell-mediated immune responses to different isolates of Bovine Herpesvirus 1 (BHV-1). Groups of high, moderate, and low responses were observed when lymphocytes from three breeds of dairy cattle were stimulated with each of the BHV-1 isolates. Proliferation of cells in the low responding group could be augmented by exogenous IL-2. The mechanism of unresponsiveness by cells from one individual whose response was not altered by IL-2 supplementation was further investigated. The patterns of response by limiting dilution frequency analysis eliminated the possibility that this individual lacked responsive cells but suggested the presence of regulatory cell interactions which resulted in the observed low proliferative response. These results show that animals exposed to the same environment can vary greatly in their ability to respond to BHV-1. At least two mechanisms may be responsible for low proliferative responses in vitro: inadequate levels of IL-2 and the presence of suppressor cells.     

A comparative study of bovine herpesvirus 1247 and equine herpesvirus 1 in ponies.

The clinical and immunological response of ponies exposed to a bovine herpesvirus isolate and equine herpesvirus 1 were compared. Each virus was inoculated into two ponies by the intranasal route. One uninoculated pony was used with each group as a contact control. The four inoculated ponies developed a mild rhinitis with an increase in rectal temperature. Virus was recovered from nasal secretions collected from the four inoculated and one contact pony. All ponies developed a serum neutralizing antibody to each virus. The data show that the two viruses are similar.     

Characterization of monoclonal antibodies to bovine herpesvirus type I, Los Angeles strain.

We produced monoclonal antibodies (mAbs) against bovine herpesvirus type 1 (BHV-1), Los Angeles strain, and then evaluated them as potential candidates for preparing diagnostic reagents. Of the 318 cell lines expressing antibodies to the virus, 60% (192) secreted IgG and 40% (126) secreted IgM. Twenty-six mAbs were selected based on enzyme-linked immunosorbent assay (ELISA) and virus neutralization (VN) titers and characterized by immunoprecipitation, immunofluorescence and immunoblots. The selected mAbs were assigned to one of four groups based on their immunoprecipitation patterns. Group A (4 mAbs) precipitated a complex of three glycoproteins with molecular weight (MW) 130 kDa, 72 kDa and 55 kDa, which presumably represented gI of BHV-1. Monoclonal antibodies of this group were highly reactive in ELISA but had low VN titers. Group B (4 mAbs) precipitated a glycoprotein with MW of 71 kDa (gIV). This group of mAbs had high VN titers. Group C (16 mAbs) precipitated a 97 kDa glycoprotein (gIII). Monoclonal antibodies of this group had high ELISA but low VN titers. Group D (2 mAbs) precipitated a double band of non-glycosylated proteins with MW of 37/32 kDa; these proteins could not be assigned to any of the antigens of BHV-1 previously described. ELISA and VN titers of this group of mAbs were low. To test the antigenic variability of the antigenic determinants which were recognized by these 4 groups of mAbs, we adapted Madin Darby bovine kidney cell-propagated BHV-1 Los Angeles strain to Crandell's feline kidney cell line. After the tenth passage in feline kidney cells, the epitopes on the 37/32 kDa peptide recognized by the mAbs group D were no longer detectable. Additional changes were noted in the electrophoretic mobility of the 130 kDa and 71 kDa glycoproteins (gI) identified by mAb of group A shifted downward. The 71 kDa glycoprotein (gIV) reactive with mAb group B and the 97 kDa (gIII) reactive with mAb group C remained stable. Since clone No. 191 of group B mAb was potent in ELISA, VN, immunoblots and immunofluorescence, and recognized an epitope which did not change under selective pressure, we feel that the mAb produced by this clone are a good candidate for the production of diagnostic reagents.     

An electron microscopic study of experimentally-induced HEV encephalitis.

A gnobiotic piglet, was inoculated intracerebrally with hemagglutinating encephalomyelitis virus (strain VW572). Mononuclear cells formed vascular cuffs and were disseminated in the brain parenchyma. A few neurons were surrounded by the same kind of cells. Virus particles morphologically similar to coronavirus particles were found in the cytoplasm of both chromatolytic light neurons and hyperchromic dark neurons. The particles were in vesicles of distended endoplasmic reticulum and in the hypertrophied Golgi apparatus.     

The scanning electron and light microscopic structure of bovine tactile hair

Bovine tactile hairs in skin samples from the lateral side of the upper lip were examined using scanning electron and light microscopy. The root of these hairs has a variable length and is surrounded by a large sinus located between the internal and the external dermal sheath. With a prominent thickness, the external dermal sheath forms the external wall of the tactile hair and contains many nerves some of which extend into trabeculae. Trabeculae projecting from the internal dermal sheath and attaching to the external dermal sheath with two or more branches are present in the entire sinus. The trabeculae are interconnected by connective tissue sheets that support the integrity of the trabecular organization. The sinus surfaces as well as trabeculae are lined by endothelia. As a result, the bovine tactile hair is truly a cavernous type of tactile hair with a well organized microscopic anatomy. Thus, the bovine tactile hair most likely plays an important role in relatively immobile and insensitive bovine lips.     

A scanning electron microscopic study of the vascular system of the bovine hind limb claw

The vascular organisation of the hind limb claws of clinically normal cattle was studied using scanning electron micrographs of plastic corrosion casts. The microvasculature of the dermal papillae and lamellae was demonstrated. A consistent vascular pattern was observed in the dermal papillae of the periople, coronary margin, sole and bulb. Ultrastructurally, the papillar vasculature consisted of a central artery and vein surrounded by a fine network of interconnected capillaries and venules. Arteriovenous anastomoses, connecting the central artery and vein, and peripherally situated arteriovenous loops were recognised in the terminal papillae of the white zone only. Arteriovenous anastomoses, focal enlargements and distension of the capillary bed were located within the interior half of the dermal lamellae. Numerous arteriovenous anastomoses were present throughout the entire dermis of the claw and were situated predominantly at the base of the dermal papillae and lamellae. The anastomoses and focal capillary enlargements may influence the blood flow in normal conditions and may play a significant role in the pathophysiology of bovine laminitis and sole ulceration.     

Frequency of occurrence of viruses associated with respiratory tract disease of cattle in Oklahoma: serologic survey for bovine herpesvirus DN599.

Serum samples from 351 Oklahoma cattle were tested for antibodies to DN599 virus by the indirect fluorescent antibody test. Seven cattle (approximately 2%) were seropositive for this virus.     

Experimental hamster enteritis: an electron microscopic study.

Hamster enteritis (HE) was experimentally produced in weanling hamsters by orally inoculating healthy hamsters with suspensions of ilea obtained from hamsters with HE. Control groups of hamsters were inoculated orally with suspensions of ilea from healthy hamsters. Electron microscopy was done on ilea from 6 control hamsters, 31 hamsters with experimentally produced HE, and 4 hamsters with naturally occurring HE. Ultrastructural changes were not observed in the absorptive epithelium of control animals. Two different intracytoplasmic bacterial organisms were observed in epithelial cells of hamsters with experimentally produced HE. Organisms that were observed early in the disease process were identified as Escherichia coli. Organisms ultrastructurally similar to Campylobacter spp were observed later in the disease and were only within hyperplastic epithelial cells. The hyperplastic ileal epithelium of hamsters with naturally occurring HE contained campylobacter-like organisms.     

Development of a polymerase chain reaction and restriction typing assay for the diagnosis of bovine herpesvirus 1, bovine herpesvirus 2, and bovine herpesvirus 4 infections.

A multiplex polymerase chain reaction (PCR) method coupled with a restriction analysis of PCR products (PCR with restriction fragment length polymorphism) was developed for the simultaneous detection of bovine herpesvirus 1, bovine herpesvirus 2, and bovine herpesvirus 4 infections. The specificity, sensitivity, and practical diagnostic applicability of this method were evaluated. This assay may be also adapted to the diagnosis of suid herpesvirus 1 and equine herpesviruses 1 and 3 and could become a powerful diagnostic tool.