Pharmacologic and toxicologic properties of humic acids and their activity profile for veterinary medicine therapy

Humic acids derive from a class of natural substances in humic substances. The chemical properties of certain defined humic acid products enable their application in diseases of the digestive system of mammals when combined with metabolic disorders, especially in rearing age. The simple administration (via feed), their exceptional safety and the absence of side effects (e. g. allergy, resistance) as well as no residue formation in animal derived products allow a broad application of these substances in veterinary medicine, even when regarding ecotoxicological aspects.     

Resistance to triforine: A nonexistent problem?

Permanent resistance to triforine in Cladosporium cucumerinum was obtained after UV irradiation of spore suspensions and selection of resistant strains in the presence of triarimol or triforine. About 50 strains were examined for mycelial growth on malt agar, viability upon routine subculturing, growth and inhibition by triforine in thin-layer chromatographic bioassays, and virulence towards cucumber seedlings. In addition, sporulation and spore germination, as well as effects of ergosterol were investigated in a restricted number of strains. Although all strains differed considerably from each other in these characteristics, they were quite similar in a reduced overall-fitness. These triforine-resistant mutants of C. cucumerinum might, therefore, have a greatly reduced chance of survival in competition with the wild-type strain, as was actually demonstrated for two strains in greenhouse experiments with mixed-inoculations of cucumber seedlings. Such a reduced chance of survival might also explain why under practical conditions development of resistance to triforine has not been observed yet.The triforine-resistant C. cucumerinum strains showed cross-resistance to a number of systemic fungicides known or supposed to inhibit ergosterol biosynthesis, viz. Denmert, fenarimol, imazalil, triadimefon and triarimol. The practical implications and potential hazards of such cross-resistance for the use of already established fungicides are discussed.     

The digestibility of crude nutrients in horses. 1. Methods of determining digestibility

The ascertainment of the digestibility of crude nutrients in rations for horses requires the inclusion of movement-based performance as influencing factor. Under the consideration of this requirement, indicator methods gain significance in comparison with the standard method of collecting faeces completely. In two experiments the digestion of the crude nutrients in rations for horses was tested according to the standard method (complete collection of faeces) and the indicator method 4 N HCl-insoluble ash). According to the indicator method the digestibility of crude nutrients was slightly higher, the differences to the values of the standard method were statistically insignificant. Under consideration of the inhomogeneity of horse faeces and the possibility of doing without the complete collection of faeces, the indicator method with 4 N HCl-insoluble ash is to be preferred to the standard method. The analysis of air-dried faeces connected with the indicator method resulted in improved reproducibility due to the significantly better homogeneity. A recommendation is made for carrying out digestibility experiments with horses, in which the 4 N HCl indicator method is used.     

Characterization of different wood samples using a new combined method of evolved gas analysis and pyrolysis–gas chromatography/mass spectrometry

Evolved gas analysis (EGA) in combination with analytical pyrolysis is presented as a new technique for rapid investigations into wooden materials. In a first analytical step, the thermal behaviour of three wood samples was determined by EGA. Based on these results, analytical pyrolysis–gas chromatography/mass spectrometry was carried out to investigate the influence of wood-specific thermal behaviour and the chemical composition on the results. Furthermore, the influence of sample mass and particle size was examined.     

IMAGING DIAGNOSIS: FIBRODYSPLASIA OSSIFICANS PROGRESSIVA IN A CAT

A 1‐year‐old female cat was presented for progressive alopecia, gait abnormalities, and stiffness. Radiography demonstrated multiple calcified lesions within the soft tissues of the cervical and thoracic spine, shoulder, and limbs. Postmortem computed tomography provided more detailed information on the distribution, pattern, and extension of lesions. In addition, computed tomography helped guide sample selection for histopathology. The final diagnosis was fibrodysplasia ossificans progressiva. This is a rare disorder of unknown etiology, characterized by fibrosis and heterotopic bone formation in connective tissues. To the authors’ knowledge, this is the first report describing this disease in a European cat.     

Detection of bovine herpesvirus type 4 antibodies and bovine lymphotropic herpesvirus in New Zealand dairy cows

AIM: To detect the presence of bovine herpesvirus (BoHV) type 4 in New Zealand dairy cows with clinical metritis.

METHODS: Serum samples taken from 92 dairy cows with clinical metritis, each from a different farm, were tested for the presence of antibodies against BoHV-4 using a commercially available, indirect ELISA. Peripheral blood mononuclear cells (PBMC) were collected from 10 BoHV-4 seropositive cows, and PBMC were examined by a pan-herpesvirus nested PCR to detect herpesvirus. PCR products were sequenced directly and a proportion of the PCR products were cloned and sequenced to identify the virus present.

RESULTS: Antibodies to BoHV-4 were detected in 23/92 (25%) serum samples. The pan-herpesvirus PCR was positive in 8/10 PBMC samples. Cloning and sequencing identified that all of the eight PCR-positive PBMC contained bovine lymphotropic herpesvirus (BLHV); no BoHV-4 DNA was detected.

CONCLUSIONS: This study reports the finding of the presence of apparent antibodies to BoHV-4, and BLHV DNA in New Zealand dairy cows affected by metritis.

CLINICAL RELEVANCE: Bovine herpesvirus type 4 and BLHV are reported to have the potential to cause reproduction failure in cows. This is the first report of apparent BoHV-4 antibodies, and BLHV in New Zealand. The importance and epidemiology of these viruses in cattle in New Zealand requires further investigation.     

Oral vaccination of brushtail possums (Trichosurus vulpecula) with BCG: immune responses,persistence of BCG in lymphoid organs and excretion in faeces

AIMS: To determine immune responses, and the localisation and persistence of Mycobacterium bovis bacille Calmette-Guérin (BCG) in gut-associated lymphoid tissues (GALT) and other organs in possums vaccinated orally with lipid-formulated BCG vaccine. To determine the duration of excretion and longevity of survival of BCG in the faeces of vaccinated animals.

METHODS: Possums (n=28) were vaccinated with lipid-formulated BCG (1 x 10 ⁸ colony forming units (cfu) of formulated BCG) by the oral route. Control possums (n=17) were fed oral bait pellets containing formulation medium only. Possums were sacrificed at 3 days and at 1, 3, 6 and 8 weeks after vaccination or ingestion of bait. Proliferation responses to bovine purified protein derivative (PPD) were measured in lymphocytes from blood and mesenteric lymph nodes (MLN) and samples of lung, spleen, liver, MLN and Peyer's patches (PP) were cultured for the presence of BCG. The number of BCG organisms excreted in faeces and the duration of excretion were determined in eight vaccinated possums and eight control possums over a 3-week period. In a separate experiment, a further six possums were vaccinated with oral BCG vaccine (5–10 x 10 ⁸ cfu BCG/possum) and their faeces collected over 48–72 h, for culture of BCG. The longevity of survival of BCG in these faeces was determined by storing faecal samples (n=12) under three different conditions: in an incubator (22.5°C), and conditions which simulated the forest floor and open pasture. A proportion (1–2 g) of these faecal samples was collected after storage for 1, 3, 5, 8 or 20 weeks, and cultured for BCG.

RESULTS: Possums vaccinated orally with BCG vaccine showed strong proliferation responses to bovine PPD in peripheral blood lymphocytes at 6–8 weeks post-vaccination (p.v.). Positive lymphocyte proliferation assay (LPA) responses to bovine PPD were first evident in MLN at 3 weeks p.v. BCG was cultured from MLN and PP in a proportion of animals at 3–8 weeks p.v. BCG was not cultured from sections of spleen, lung or liver at any time p.v. BCG was recovered in low to moderate numbers from the faeces of vaccinated possums for up to 7 days, and maximal numbers were cultured in faeces collected 48–72 h p.v. After storage for 1 week, BCG was cultured from all faecal samples placed in the incubator and from a proportion of faeces exposed to conditions similar to those on the forest floor and pasture. With the exception of one faecal sample stored under forest floor conditions which was culture-positive for BCG at 3 and 5 weeks, BCG was not cultured from any other faecal sample stored for more than 1 week.

CONCLUSIONS: Ingestion of oral BCG vaccine by possums was associated with the development of strong cell-mediated immunity in both blood and MLN. Following oral vaccination with BCG, the organisms were localised and persisted in GALT but did not spread to the spleen, liver or lungs. BCG was shed in low to moderate numbers in the faeces for up to 7 days p.v. The viability of BCG excreted in faeces decreased rapidly, particularly when faeces were exposed to an open pasture environment. Oral vaccination of possums with formulated BCG is unlikely to result in undue contamination of the environment with BCG.