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.     

Impact of bovine somatotropin administration beginning at day 70 of lactation on serum metabolites, milk constituents, and production in cows previously exposed to exogenous somatotropin.

Metabolic and production responses are reported for 72 cows treated with bovine somatotropin (BST) for 30 days starting at day 70 of lactation. Of these 72 cows, 48 had been exposed in the preceding lactation to long-term treatment with BST at 3 dosages and 24 (controls) had not been given BST. Approximately half of the cows in each group were parity-2 cows, the rest were older. Comparisons between groups were made separately for parity-2, and older cows. Analyses, using pretreatment values of each variable as a covariate, indicated that older cows, but not parity-2 cows, significantly (P less than 0.05) increased milk production during treatment. Parity-2 cows, however, had a significantly higher milk fat percentage than controls following treatment. Cows treated with 51.6 or 86 mg BST/d in both parity groups had significantly higher serum-free fatty acids than controls. Estimated net energy balances were significantly lower for older treated cows, but did not significantly differ from controls for parity-2 treated cows. Older cows in the 86 mg of BST/d group tended to have higher concentrations of blood glucose than did older control-group cows. Treatment with BST did not significantly increase serum ketone concentrations in any group of animals, and none of the cows developed clinical ketosis during this period. Estimated net energy balance (ENEB) during treatment was a significant (P less than 0.05) covariate for free fatty acid concentrations in older cows and for milk fat percentage in parity-2 cows. Covariate adjusted analyses, using ENEB during treatment as a covariate, indicated that lipolytic stimuli already acting may be enhanced by treatment with BST, but a negative energy balance was not a necessary precondition for free fatty acid concentrations to increase following somatotropin treatment. Similarly, milk fat percentages for parity-2 treated cows were significantly (P less than 0.05) higher during treatment than controls when ENEB during treatment was used as a covariate. Increased milk fat concentrations in parity-2 treated cows were not associated with significant increases in the ratio of C18:C4-10 milk fatty acids, indicating that increased milk fat resulted from either an increase in incorporation of C18 fatty acids into milk fat coupled with an increase in de novo mammary synthesis of C4-10 milk fatty acids or an increase in C12-16 fatty acids that may arise either from increased tissue mobilization, from diet, or from de novo mammary synthesis.     

Response time of broiler chickens to cimaterol: meat tenderness, muscle composition fiber size, and carcass characteristics.

The response time to cimaterol (CIM), a beta-adrenergic agonist, by broiler chickens for carcass characteristics, muscle composition, muscle fiber size, catheptic enzyme activity, and tenderness was determined. Two trials were conducted in which chickens were fed a control diet (CON) containing 0 ppm of CIM or a diet containing 1 ppm of CIM. Trial 1 consisted of 55, 31-d-old broiler chickens individually fed for up to 48 h. At 0, 6, 12, 18, 24, and 48 h, five CON and five CIM-fed chickens were killed. Trial 2 consisted of 160, 33-d-old broiler chickens group-fed for up to 14 d. At 2, 4, 6, 8, 10, 12, and 14 d, 10 CON and 10 CIM-fed chickens were killed. The breast muscle (BM) and leg muscle (LM) weight, cathepsin B and L activities, DNA, RNA, and protein concentration, and BM shear force value (SFV) were measured in both trials. Thigh muscle (TM) SFV were measured in Trial 2 only. Fiber size of BM was measured (five birds per treatment) at d 2, 6, 10, and 14. In Trial 1, BM weight and SFV were lower in CIM-fed birds at 6 h (P less than .05). In Trial 2 BM SFV were higher at d 8 (P = .06) and d 10 (P less than .05) in CIM-fed chickens. The SFV of CIM-fed chickens were higher at d 4, 8, 10, 12, and 14 (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Control of Johne's disease in four commercial dairy herds in Iowa

A 6-year study was conducted in 4 dairy herds in Iowa in which Johne's disease was diagnosed previously. Fecal specimens were collected at 6-month intervals from animals 2 years of age and over for mycobacteriologic examination. Serum samples were obtained at 3-month intervals and tested by enzyme-linked immunosorbent assay (ELISA). The antigen used in the ELISA was a potassium chloride extract of a field strain of Mycobacterium paratuberculosis. The ELISA reactions were observed in 87% of the cows from which M. paratuberculosis was isolated. Dairy producers that participated in the Johne's control program reported reduced economic losses. Increased income was attributed to improved milk production, increased value of vaccinated animals sold as replacements to other dairy herds in which Johne's disease had been diagnosed, and the increased market value of slaughter animals removed from the herd.     

Ultrasonography as an aid to controlled breeding in the llama (Lama glama).

An ultrasonic linear array scanner with a transrectal probe was used to observe ovarian and uterine changes associated with the reproductive cycle in llamas. 'Waves' of follicular development and regression occurred in unstimulated females, during which the dominant follicle reached a maximum size of 9 to 13 mm; both ovaries were equally active. Ovulation was induced by mating in 80 per cent of cases, and when mating was accompanied by the administration of human chorionic gonadotrophin the ovulation rate increased to 90 per cent and the time to ovulation decreased from two to three days to one to two days. Some spontaneous ovulations occurred. Corpora lutea reached a maximum size of 12 mm (non-pregnant) or 14 mm (pregnant) after seven or 16 days, respectively. The lifespan of the corpus luteum was approximately 11 days in non-pregnant llamas and the regression time was advanced by the administration of prostaglandin or embryo recovery. Pregnancy could be diagnosed as early as 19 days after mating.     

Isoleucine requirements of the chicken: the effect of excess leucine and valine on the response to isoleucine.

1. Three experiments were designed to determine the response of broiler chickens to dietary isoleucine, and to quantify the antagonistic effects of excess leucine and valine on this response. 2. A dilution technique was used to measure the responses in growth rate and food intake to a range of diets differing in their isoleucine concentrations. A summit diet was formulated to contain isoleucine at 1.14 times the requirement and with leucine (1.76 times the requirement) and valine (1.87 times the requirement) at the minimum possible concentrations, given the ingredients available. A dilution mixture, devoid of protein, was formulated to correspond in all respects, other than in amino acid content, to the summit diet. These two basal diets were blended in different proportions to give a range of diets of decreasing isoleucine and protein content. 3. In experiment 1 the response was measured to isoleucine with leucine and valine remaining in the same proportion to isoleucine throughout the range of diets fed. In experiments 2 and 3, however, L-leucine and L-valine were added to the diets either singly or in combination to give 6 isoleucine concentrations and 3 ratios of each of leucine and valine to isoleucine. 4. Weight gain decreased as the isoleucine content of the diet was reduced, whereas food intake of broilers fed on the marginally deficient diets increased to a maximum and then decreased. FCE decreased curvilinearly as the isoleucine concentration in the food decreased, reflecting a concomitant change in the fat content of the broilers. 5. It is possible that the amount of dietary isoleucine assumed to be available to the broilers in these experiments was overestimated by hydrolysing the food samples for 72 h, and the doubt thus created makes an estimate of the efficiency of retention of isoleucine suspect. 6. Excess valine had no effect on the response to isoleucine, whereas an increase in the leucine to isoleucine ratio depressed food intake and hence weight gain, but only at the lowest concentrations of isoleucine. 7. If the food content of isoleucine is sufficient to meet the requirements of the broiler, relatively large excesses of leucine, of valine, or of both will not depress growth.     

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.     

Isotypic antibody responses in cattle infected with Haemophilus somnus

Bovine antibody responses to Haemophilus somnus were compared on the basis of clinical and bacteriological findings. Serum IgG1 and IgM antibody titres were significantly increased in clinically normal cattle that were bacteriologically positive for H somnus from the nasal or vaginal mucosae compared with clinically normal, negative cows. IgG2 titres did not differ significantly between these two groups. However, IgG2 antibody was significantly higher in animals with H somnus disease (pneumonia or abortion) than in clinically normal cattle (whether bacteriologically positive or negative), while IgG1 and IgM titres did not differ between diseased and bacteriologically positive, clinically normal cattle. These antibody trends were duplicated in experimental H somnus abortion or pneumonia, with the greatest response occurring within the IgG2 subclass. Cattle vaccinated systemically with killed whole H somnus produced a predominant IgG2 response with minimal IgG1 and IgM responses. These results demonstrate that IgG2 antibody is consistently elevated in H somnus disease, and suggest that this response may be useful in discriminating diseased from asymptomatic cattle.