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.     

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.     

Hepatitis contagiosa canis (H.c.c.)--2 cases in Austria

Two cases of H.c.c. which occurred in winter 1987 in Vienna are described. Case one was a female Chow-Chow, 8 weeks of age, that died from the peracute form of the disease. The diagnosis was confirmed by histology and direct immunofluorescence. Case two, a 9-month old female Kuvacz, showed clinical signs of the subacute form of H.c.c. She was hospitalized and therapy was successful. The disease was diagnosed by the typical clinical signs and the raise of antibodies in paired serum samples. Etiology, clinical signs and immunology of H.c.c. are discussed.     

Differentiation of hog cholera and bovine virus diarrhoea viruses in pigs using monoclonal antibodies

Monoclonal antibodies against hog cholera and bovine viral diarrhoea viruses were assayed on organ tissue sections of experimentally infected animals. The animals had been infected simultaneously with both viruses. The antibodies were tested using an indirect immunofluorescence test and an indirect enzyme immunoassay with a biotin/streptavidin/peroxidase detection system. A polyclonal hyperimmune serum was used as a control in direct immunofluorescence tests. Both techniques based on monoclonal antibodies were more sensitive and more specific than the conventional test, the enzyme immunoassay being more sensitive than the immunofluorescence test. Small amounts of BVD viral antigen were demonstrable with monoclonal antibodies in most organ tissues.     

Management of rectal injuries

Diagnosis, evaluation, and management of the various grades of rectal tears is discussed. Surgical techniques, which include direct closure, diverting colostomies, and placement of temporary rectal liners, are detailed. Also, rectal prolapses and various methods of repair are outlined.     

Practice versus family: the great balancing act

Dairy practice often requires substantial time commitments. Those veterinarians with families may find that if they do not actively budget some time for spouse and children, the family unit may suffer. A successful practice and meaningful family life are both possible if the doctor or doctors involved are willing to work for their achievement.     

Cytological, Immunocytochemical and Ultrastructural Study of the Adenohypophyseal Pars Distalis of the Kid (Capra hircus): The TSH cell

The structure and ultrastructure of the adenohypophyseal pars distalis in kids of the Murciano-granadiana breed (18 males and 12 females), aged 2-3 months and with an average carcass weight of 8 kg has been studied. Techniques of staining (Tetrachrome Herlant's, and Cleveland-Wolfe) and histochemistry (PAS, PAS-OG and BA-PAS-OG) contrasted with immunolabelling (PAP) have been used. In addition an ultrastructural study has been made and nucleus and cytoplasm, secretory granules, mitochondria and lysosomes have been measured with a semiautomatic image analyzer (IBAS-1). TSH cells are found in sagittal section in the anterior area and in an antero-caudodorsal band, and transversally in the ventral and medial region; they are 6% and their average size is 14.15 microns. Ultrastructurally they are a single cellular type with spherical granules whose size is 195 nm in males and 149 in females; these granules are characterized by a clear halo and a variable electronic density. The rough endoplasmic reticulum is found as slightly enlarged vesicles with a homogeneous and moderately electro-dense content or in concentric stratifications.     

Evaluation of three strip tests for measuring electrolytes in mares' pre-partum mammary secretions and for predicting parturition

Three strip test kits which measure calcium and/or magnesium in mares' mammary secretions were compared and assessed for their accuracy in:- 1) measuring these electrolytes compared to standard laboratory measurements, and 2) predicting the time of parturition. Daily samples of mammary secretions were obtained pre-partum from 35 Thoroughbred mares and allocated a score. Measurement of mammary secretion calcium and/or magnesium by the strip tests and laboratory methods correlated significantly (P less than 0.001). The results obtained for prediction of parturition were analysed by two methods. Method i) indicated that the majority of mares had a maximum score on the day of birth. However, the second method of analysis included all prepartum scores and this demonstrated that maximum scores occurred on more occasions prior to the day of birth than on the day of birth itself. The number of nights spent attending mares, and the percentage of occasions when foaling was missed, were also calculated for given strip test scores. The results indicate that the test kits are not particularly accurate in predicting time of parturition although they are a reliable means of measuring mammary secretion calcium and/or magnesium. It is suggested that the test kits may be more helpful by indicating when it is not necessary to attend mares at night.     

Ultrastructure of sarcocysts from water buffalo in India

The ultrastructure of sarcocysts of macro- and microscopic species of Sarcocystis was compared from naturally infected water buffalo from India. Grossly visible sarcocysts had walls consisting of cauliflower-like villar protrusions, typical of S. fusiformis. The sarcocyst wall of the microscopic species of Sarcocystis was 6.4 microns thick and consisted of tightly packed conical villar protrusions that were 9.6 microns long and 3.7 microns wide at the base. At approximately 3 microns above the base, the distal two-thirds of the villar protrusion became conical shaped and was bent laterally at an angle of 45 degrees to the sarcocyst surface. The granular layer beneath the villar protrusions was 0.9 microns thick. In S. levinei the granular layer was 1.9 microns thick, the villar protrusions were narrow and it had a highly undulating primary cyst wall. Whether the microscopic S. levinei-like sarcocysts of Indian and Malaysian water buffalo are distinct species of Sarcocystis will require further investigation.