Development of Immune Response to Experimental Bovine Trichophyton vervucosum Infection

Abstract— Cell mediated and humoral immune responses to experimental Trichophyton verrucosum infection were assessed by sequential cutaneous biopsies, antibody assessments and microscopic monitoring of fungal presence. Histopathologic examination showed the accrual of lymphocytes and other inflammatory cells in the dermis of infected sites. Immunoperoxidase staining of frozen sections with monoclonal antibody preparations revealed an influx of macrophages, BoCD4+ and B0CD8+ lymphocytes and γδ T cells from the 5th day to the 33rd day of infection. A moderate influx of B cells was observed. Protein G-colloidal gold staining revealed the presence of immunoglobulins in the dermis and superficial epidermal layers. Trichophyton specific serum antibodies appeared between days 33 and 55. Microscopic assessment of infected tissues revealed an increase in T, verrucosum elements (mycelium and ectothrix spores) from days 19 to 55. Fungal elements in infected areas did not decrease until after both humoral and cell mediated elements of the immune response were established. These responses imply a combination of cell mediated and humoral events were associated with T. verrucosum immunity and clearance in the calf. Résumé— Le réponse immunitaire humorale et cellulaire a une infection expérimentale àT. verrucosum a été appréciée par des biospsies cutanées successives, des dosages d'anticorps et la recherche microscopique de champignons. L'examen histopathologique a montré un afflux de lymphocytes et d'autres cellules inflammatoires dane le derme des sites infectés. Les marquages en immunopéroxydase par un anticorps monoclonal de coupes congelées a montré un influx de macrophages, lymphocytes BoCD4+ et BoCD8+ et des cellules T γδ, du 5e au 33e jour de l'infection. Un marquage par une protéine G—or colloidal a révélé d'immunogolglobulines dans le derme et les couches supéerficielles de l'épiderme. Les anticorps spécifiques de Trichophyton sont apparus entre 33 et 55 jours. L'examen microscopique des tissus infectés a révélé une augmentation du nombre d'éléments de T. verrucosum (mycelium et spores ectothrix) du 19e au 55e jours. Les éléments fongiques dans les zones infectées n'ont pas diminué avant que les réponses humorales et cellulaires ne solent établies. Ces résponses impliquent qu'une coopération des réponses humorales et cellulaires étaient associées dans l'immunité et la défense contre T. verrucosum. Zusammenfassung— Die zellvermittelten und humoralen Immunantworten auf die experimentelle Infektion mit T. verrucosum wurden durch eine Serie von Hautbiopsien, Antikörperuntersuchungen und mikroskopischer Untersuchung auf das Vorhandensein von Pilzen ausgewertet. Die histopathologische Untersuchung zeigte eine Ansammlung von Lymphozyten und anderen Entzündungszellen in der Dermis der infizierten Stellen. Die Immunperoxidasefärbung der Gefrierschnitte mit monoklonalen Antikörperzubereitungen zeigte einen Influx von Makrophagen, BoCd4-und BoCD8-Lymphozyten und gamma-delta-T-Zellen vom 5. bis zum 33. Tag der Infektion. Es wurde auch ein mäßiger Influx von B-Zellen beobachtet. Die Protein-G-kolloidale Goldfärbung zeigte die Anwesenheit von Immunglobulinen in der Dermis und den oberflächlichen epidermalen Schichten. Trichophyton-spezifische Serumantikörper traten zwischen Tag 33 und 55 auf. Die mikroskopische Untersuchung infizierter Gewebe zeigte eine Zunahme von T. verrucosum-Bestandteilen (Myzel und exktothrixe Sporen) vom Tag 19 bis 55. Pilzteile in infizierten Bereichen verminderten sich weder, nachdem humorale, noch nachdem zellvermittelte Elemente der Immunantwort auftraten. Diese Reaktionen deuten an, daß eine Kombination von zellvermittelten und humoralen Vorgängen im Zusammenhang mit T. verrucosum-Immumtät und Abheilung beim Kalb vorliegt. Resumen Por medio de biposias cutáneas secuenciales, medida de anticuerpos y exámen microscópico de presencia de hongos, se estudió la respuesta inmunitaria de tipo humoral y celular producida por la infección experimental con T. verrucosum. El exámen histopatológico reveló la presencia de agregación de linfocitos y otras células inflamatorias procedentes de la dermis de los puntos infectados. La tintura por medio de inmunoperoxidasa de las secciones congeladas, con preparaciones monoclonales de anticuerpos, demostró un aflujo de macrófagos BoCD4 + y BoCD8 + linfocitos y linfocitos, Tαδ, desde el quinto hasta el día 33 la infección. También se observó un aflugo moderado de linfocitos B. La tintura aúrica de proteina coloidal G reveló la presencia de inmunoglobulinas en al dermis y capas superficiales de la epidermis. Los anticuerpos específicos para la especie Trichophyton aparecieron entre los días 33 y 55. El exámen microscópico de los tejidos afectados demostró un incremento, de los elementos füngicos T. verrucosum (micelio y esporas exótricas) desde los días 19 al 55. Los elementos fúngicos en áreas infectadas no disminuyeron hasta después del establecimiento de ambos tipos de respuesta inmunitaria, humoral y celular. Estas respuestas implican que la combinación de ciertos fenómenos de inmunidad celular y humoral, están relacionados con la desaparición y la inmunidad de la infección producia por T. verrucosum en el ternero.     

Hematological parameters and visceral lesions relationships in rabbit viral hemorrhagic disease.

Twenty rabbits were inoculated with a suspension of Viral Hemorrhagic Disease virus. Hemostatic functions were assessed every sixth hour from 6 to 60 hours post-inoculation. Tissue samples obtained at the same intervals allowed the study of the development of lesions throughout the experiment. Biological signs of Disseminated Intravascular Coagulation (DIC) were detected on and after 30 h post-inoculation and consisted of prolonged One Stage Prothrombin Time and Activated Partial Thrombin Time, the decrease of factors V, VII, and X and high levels of soluble fibrin monomer complexes and D-dimers. A reduction of thrombocyte numbers, heterophils and lymphocytes was associated. The close association of DIC and necrotizing hepatitis lesions suggested the hepatic lesions to be the most important DIC triggering factor. Other mechanisms are discussed.     

Specificity of the carbon immunoassay (CIA) test for the diagnosis of Toxoplasma infections.

A rapid and low cost procedure, the carbon immunoassay (CIA) test, was evaluated for the diagnosis of Toxoplasma gondii infections. Using a closely related parasite (Besnoitia jellisoni) as antigen, and homologous or heterologous immune sera, it was demonstrated by light and electron microscopy that CIA is a very reliable and specific test. As it is neither expensive nor time-consuming, it can be recommended for general and routine laboratory use.     

Transrectal ultrasonography for the evaluation of stallion accessory sex glands.

This article reviews the capabilities of transrectal ultrasonography for determining the distribution of fluid and tissue within stallion accessory sex glands. Emphasis is placed on describing the normal ultrasonographic appearance of the accessory sex glands, excurrent ducts, and pelvic urethra of stallions during rest, after teasing, and after ejaculation and using this information to detect glandular abnormalities.     

Oxidative damage to the membrane of canine erythrocytes with inherited high Na, K-ATPase activity.

Oxidative damage to the membrane in canine erythrocytes with inherited high Na, K-ATPase activity (HK cells) was compared with that in normal canine cells (LK cells). When 30 mM beta-acetylphenylhydrazine (APH) was applied to HK and LK cells, lipid peroxidation and hemoglobin denaturation occurred. Lipid peroxidation determined from malondialdehyde (MDA) formation was significantly lower in HK than in LK cells so far as endogenous glutathione (GSH) concentration was maintained at appropriate levels. With the depletion of GSH, MDA formation was accelerated and difference between HK and LK cells was not significant. Denatured hemoglobin bound to the membrane protein was less in HK than in LK cells. During incubation with APH, osmotic fragility increased markedly in LK cells, while HK cells showed very little change. The amounts of total lipid, total and free cholesterol, glycolipid, phospholipid and fatty acids were essentially the same in both cell types. Fatty acid compositions showed very small differences. The membrane of HK cells thus appear to have greater protection against oxidative damage induced by APH, owing to the presence of excess GSH in HK cells. The capability of HK cells to withstand oxidative damage would not be due to differences in membrane lipid compositions.     

In situ radiometric mapping of soil erosion and field-moist bulk density on cultivated fields

Abstract. Recent developments in in situγ ray spectrometry offer a new approach to measuring the activity of radionuclides such as ¹³⁷Cs and ⁴⁰K in soils, and thus estimating erosion or deposition rates and field moist bulk density (ρ_m). Such estimates would be rapid and involve minimal site disturbance, especially important where archaeological remains are present. This paper presents the results of a pilot investigation of an eroded field in Scotland in which a portable hyper pure germanium (HPGe) detector was used to measure γ ray spectra in situ. The gamma (γ) photon flux observed at the soil surface is a function of the ¹³⁷Cs inventory, its depth distribution characteristics and ρ_m. A coefficient, Q_Cs, derived from the forward scattering of ¹³⁷Cs γ ray photons within the soil profile relative to the ¹³⁷Cs full energy peak (662 keV), was used to correct the in situ calibration for changes in the ¹³⁷Cs vertical distribution in the ploughed field, a function of tillage, soil accumulation and ρ_m. Based on only 8 measurements, the agreement between in situγ ray spectrometry and soil sample measurements of ¹³⁷Cs inventories improved from a non significant r²=0.05 to a significant r²=0.62 (P<0.05). Erosion and deposition rates calculated from the corrected in situ¹³⁷Cs measurements had a similarly good agreement with those calculated from soil cores. Mean soil bulk density was also calculated using a separate coefficient, Q_K, derived from the forward scattering γ photons from ⁴⁰K within the soil relative to the ⁴⁰K full energy peak (1460 keV). Again there was good agreement with soil core measurements (r²=0.64; P<0.05). The precision of the in situ¹³⁷Cs measurement was limited by the precision with which Q_Cs can be estimated, a function of the low ¹³⁷Cs deposition levels associated with the weapons testing fallout and relatively low detector efficiency (35%). In contrast, the precision of the in situ ρ_m determination was only limited by the spatial variability associated with soil sampling.     

Differences in Reproductive Performance, Embryo Development, Interferon-tau Secretion by the Conceptus and Luteal Function in Ewe Lambs Synchronized in Oestrus before or after the Spontaneous Onset of Luteal Activity Preceding Puberty

In mid-September, 1 month before the insertion of intravaginal pessaries to induce sexual activity, blood samples were collected every 4 days from 16 ewe lambs aged 7 months, in order to determine the incidence of ovulations by measurement of plasma progesterone concentrations. It has been studied whether the response to a progestagen treatment of ewe lambs apparently close to puberty could be modified by the onset of the ovarian events preceding puberty. The effect of the presence or absence of ovulations prior to progestagen treatment on the potential reproductive performance (fertility, litter size and fecundity), embryo development [embryo quality and interferon-tau (IFNτ) secretion], luteal function (progesterone secretion in vitro ) and endometrial progesterone content was studied in seven ovulating (Ov+) and nine nonovulating ewe lambs (Ov−) on day 14 after mating. The best potential reproductive results were obtained with Ov+ animals, although these differences could not be initially attributed to either different progesterone secretion in vitro or concentration of endometrial progesterone. Irrespective of the experimental groups, secretion of progesterone by luteal tissue from ewe lambs with normal embryos was significantly greater (p<0.05) than that of animals with abnormal embryos or with no embryos. Normal embryos secreted a higher amount of IFN-τ than those embryos classified as abnormal (p<0.07). In conclusion, ewe lambs which exhibit luteal activity before puberty have the highest levels of reproductive performances after a progestagen treatment. Corpora lutea from ewe lambs with normal embryos had higher rates of progesterone secretion in vitro and their embryos had a higher IFN-τ production by the embryos, indicating greater capacity for subsequent development.     

Dietary Lysine Requirement of Juvenile Red Drum Sciaenops ocellatus

The dietary lysine requirement of juvenile red drum Sciaenops ocellatus was reevaluated in two separate experiments. In the first experiment, lyophilized red drum muscle was combined with an L-form crystalline amino acid premix to yield diets containing approximately 35% protein on a dry-matter basis. Gradations (0.25%) of L-lysine HCl were added to the basal diet containing approximately 1.0% lysine. Each diet was fed to triplicate groups of juvenile red drum initially weighing 6–7 g for 8 weeks. Based on growth and feed efficiency data, the lysine requirement (±SE) was determined to be 1.55% (±0.079%) of dry diet or 4.430% of dietary protein. In a second experiment, the lysine requirement of red drum was investigated with diets containing 35% intact protein from zein and red drum muscle. Each of the zein-based diets containing incremental levels of lysine was fed to triplicate groups of juvenile red drum initially weighing 1–2 g for 8 weeks. A crystalline amino acid test diet serving as a control in this experiment significantly outperformed the zein-based diets, and palatability of those diets was questionable due to an obvious change in feeding behavior of the fish. Due to the inferior performance of the fish fed the zein-based diets, the authors were not successful in determining a requirement with these diets. Thus, a lysine requirement of 1.55% of dry diet (4.43% of dietary protein) as determined in the first experiment is recommended for juvenile red drum.     

Legal regulations and practical implementation of inspection of slaughter animals and their meat, concerning fur-bearing game

Meat regulations also involve game inspection. This publication deals with the regulations concerning furred game intended for slaughter and furred game hunted for food.     

Spontaneous regression of bovine cutaneous leukosis

Biopsies of skin from a 2-year-old heifer with spontaneously regressing dermal leukosis were examined. The heifer was not infected with bovine leukemia virus and was negative for tumor-associated antigens of enzootic bovine lymphosarcoma. By hematological standards for bovine leukemia, the heifer was positive at about 60 days post-occurrence of the disease (POD). At 53 days POD, lymphoblastic neoplastic cells in the dermis reacted with anti-T lymphocyte monoclonal antibody by the avidin biotin peroxidase complex method. The cells had intracytoplasmic clustered dense bodies under electron microscopy. From 53 to 83 days POD, figures of the transepithelial elimination (TE) against neoplastic cells and perivascular infiltration of small lymphocytes were in the dermis. Small lymphocytes reacted with anti-T lymphocyte monoclonal antibody. At necropsy there were no neoplastic lesions; there were flat lymph node-like tissues in the subcutis. Many germinal centers were seen in the lymphatic organs. Blood lymphocytes at 46 days POD were stimulated by phytohemagglutinin-P and concanavalin A. Sera, taken until 75 days POD and at necropsy, showed an inhibitory effect on mitogen-induced blastogenesis of normal bovine lymphocytes. These results suggested the existence of a spontaneous regressive mechanism against neoplastic lesions by TE and tumor immunity.