Comparative FT-Raman spectroscopy of Xanthoria lichen-substratum systems from temperate and antarctic habitats

Lichens have a primary role in the biodeterioration of rock substrata and in soil formation. In extreme Antarctic rock-face habitats, their exposed epilithic growth form makes them vulnerable to environmental stress, especially to increased UVB radiation within the late winter ozone “hole”. In this study, FT-Raman spectra of intact epilithic samples of the highly-pigmented lichen species Xanthoria elegans and Xanthoria mawsonii from a continental Antarctic habitat in central Victoria Land and Xanthoria parietina from a maritime temperate location in Scotland have been compared. Vibrational assignments have been made for their pigments, based on parietin and an anthraquinone model. Other significant spectral features are assigned to biodegradative secondary metabolites, cellulose and incorporated substratal rock particles. Differential distribution of parietin pigment bands in the profile of an X. elegans thallus is probably related to the high intensity of UV-radiation reaching its Antarctic cold desert surface environment. The significant differences in Raman spectra between epiphytic X. mawsonii and epilithic X. elegans at the same site possibly indicate different biochemical UV survival strategies. In contrast to spectra from other lichen species studied in our laboratories, the Xanthoria encrustations contain very little calcium oxalate, formed by reaction of oxalic acid with calcareous substrata. The 462 cm⁻¹ ν(SiO) band of quartz is found in the Raman spectra from both upper and lower surfaces of the lichen encrustations, confirms the physical incorporation of the substratum into the thallus.     

Measurement of glomerular filtration rate, renal plasma flow, and endogenous creatinine clearance in cheetahs (Acinonyx jubatus jubatus).

Glomerular filtration rate (GFR), renal plasma flow (RPF), and the endogenous creatinine clearance (CCr) rate were determined in 13 captive cheetahs, Acinonyx jubatus jubatus (seven females and six males, 1.5-7.5 yr of age, x = 5.02 yr), during general anesthesia with Telazol and isoflurane by measuring the urinary clearances of inulin, para-aminohipppuric acid, and endogenous creatinine, respectively. Methods to determine GFR, RPF, and endogenous CCr in captive cheetahs were evaluated, and the relationship between GFR and CCr for this species was determined. The GFR and the RPF were stable during the procedure, with mean values of 1.59+/-0.17 ml/min/kg body weight and 5.12+/-1.15 ml/min/kg body weight, respectively. Although the mean value for CCr (1.47+/-0.20 ml/min/kg body weight) was significantly less than the corresponding value for GFR, the mean difference (0.11+/-0.02 ml/min/kg weight) between the two measurements was slight, and the values were highly correlated (R2 = 0.928; P < 0.0001). The measurement of CCr in cheetahs should provide a reliable estimate of GFR, facilitating the early detection of renal disease in this species.     

The effects of levamisole on oxidative stress induced by copper intoxication in broilers

Abstract

AIM: To determine the effects of Cu and levamisole on concentrations of Cu and Fe in plasma and liver, and the effects of levamisole on lipid peroxidation induced by Cu intoxication in broiler chickens.

METHODS: In a 2×4 factorial study, 80 one-day-old Ross PM3 broiler chicks were fed diets for 21 days containing either 8 mg/kg Cu (Low Cu) or 250 mg/kg Cu (High Cu) and were treated with 0 (L0), 4 (L4), 8 (L8) or 16 (L16) mg/kg bodyweight levamisole per day from Day 7 of the study, on three consecutive days in their drinking water. This treatment was repeated three times, at 3-day intervals. On Day 21, blood samples were collected from each bird for analysis of concentrations of Cu, Fe and malondialdehyde, and activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), superoxide dismutase, catalase and glutathione peroxidase (GSH-Px). The birds were killed and liver samples collected for analysis of Cu and Fe.

RESULTS: Mean concentrations of Cu and Fe in plasma, and Cu in liver, were increased overall in the High Cu groups compared with the Low Cu groups (p<0.001). Compared with the L0 treatment group on the High Cu diet, treatments L4, L8 and L16 decreased concentrations of Cu in plasma, and L8 and L16 increased concentration of Cu in liver (p<0.05). Mean activities of AST and ALT were increased in untreated birds (L0) fed the High compared with Low Cu diets (p<0.01). In birds receiving the High Cu diet, treatments L4 and L8 decreased activities of AST, and L4 and L16 decreased activity of ALT, compared with L0 (p<0.05). The High Cu diet induced an oxidative stress characterised by increased mean concentrations of malondialdehyde and decreased activities of superoxide dismutase, catalase and GSH-Px (p<0.001). Concentration of malondialdehyde, and activities of superoxide dismutase and catalase were not changed following levamisole treatment in birds on the High Cu diet, and activity of GSH-Px was decreased by the L4 and L8 treatments compared with the L0 group.

CONCLUSIONS AND CLINICAL RELEVANCE: The results of the study suggest that treatment with levamisole might alleviate the harmful effects of Cu on the liver, as demonstrated by decreased activities of AST and ALT induced by a diet containing 250 mg/kg Cu.     

Alpha‐Linolenic Acid Supplementation in Tris Extender Can Improve Frozen–Thawed Bull Semen Quality

The study was conducted to evaluate the effects of α ‐linolenic acid (ALA) on frozen–thawed quality and fatty acid composition of bull sperm. For that, twenty‐four ejaculates obtained from three bulls were diluted in a Tris extender containing 0 (control), 3, 5, 10 and 15 ng/ml of ALA. Extended semen was incubated at 37°C for 15 min, to allow absorption of ALA by sperm cell membrane. The sample was chilled for 2 h, packed into 0.25‐ml straws and frozen in liquid nitrogen for 24 h. Subsequently, straws were thawed and evaluated for total sperm motility (computer‐assisted semen analysis), membrane functional integrity (hypo‐osmotic swelling test), viability (eosin‐nigrosin), fatty acid composition (gas chromatography) and lipid peroxidation (thiobarbituric acid‐reactive substances (TBARS)). A higher (p < 0.05) percentage of total sperm motility was observed in ALA groups 5 ng/ml (47.74 ± 07) and 10 ng/ml (44.90 ± 0.7) in comparison with control (34.53 ± 3.0), 3 ng/ml (34.40 ± 2.6) and 15 ng/ml (34.60 ± 2.9). Still, the 5 ng/ml ALA group presented a higher (p < 0.05) percentage of viable sperms (74.13 ± 0.8) and sperms with intact membrane (74.46 ± 09) than all other experimental groups. ALA concentration and lipid peroxidation in post‐thawed sperm was higher in all treated groups when compared to the control group. As such, the addition of 5 ng/ml of ALA to Tris extender improved quality of frozen–thawed bull spermatozoa.     

Compressive damage to the deep branch of the lateral plantar nerve associated with lameness caused by proximal suspensory desmitis

Objective— To describe pathologic changes in the deep branch of the lateral plantar nerve (DBLPN) of horses determined to be lame because of proximal suspensory desmitis (PSD), and to report the outcome after treatment by excision of a segment of the horses' DBLPN.
Study Design— Retrospective case series.
Animals— Adult horses (n=16).
Methods— Horses determined to be lame on one or both pelvic limbs because of PSD were treated by excision of a segment of the DBLPN, and 30 nerves were examined histologically. Owners were contacted to obtain information about the horses ≥6 months after surgery.
Results— Histologic changes suggestive of chronic nerve compression were identified in both nerves of 11 bilaterally lame horses and in the lame limb of 5 unilaterally lame horses. The nerve of the sound limb of 2 of 3 unilaterally lame horses that had bilateral nerve resection also had histologic changes compatible with nerve compression. Ten of 16 horses (62.5%) with follow-up information returned to soundness after excision of the DBLPN.
Conclusions— Pathologic changes of the DBLPN associated with compression may complicate PSD of the pelvic limbs. Excision of the nerve may resolve lameness caused by PSD.
Clinical Relevance— Horses lame because of PSD of the pelvic limb may remain lame after desmitis has resolved because of compression of the DBLPN. Excising a portion of this nerve may resolve lameness.     

Free radical oxidation products in plasma and synovial fluid of horses with synovial inflammation

SUMMARY: Free radical oxidation products, namely conjugated dienes, ultraviolet fluorescence (excitation 325 nm, emission 395 nm) and visible fluorescence (excitation 360 nm, emission 460 nm) were measured in equine synovial fluid exposed to free radicals In vitro and in the plasma and synovial fluids of horses with synovial effusions. The synovial effusions were induced by intra-articularly administered carrageenin (0.3 ml, 1%), which rarely resulted in clinical lameness. The free radicals were generated In vitro by mixtures of iron and ethylene diamine tetra acetate (Fe/EDTA) or mixtures of hypoxanthine and xanthine oxidase (HX/XO). The conjugated diene concentrations and intensity of ultraviolet fluorescence were negligible in plasma and synovial fluid specimens. No increase resulted from incubation of synovial fluids with either a free radical generating system or as a result of the induced inflammation. The intensity of visible fluorescence did not increase in specimens incubated with Fe/EDTA. However, the intensity of visible fluorescence increased in specimens incubated with HX/XO, in synovial effusions induced by carrageenin, in plasma and in synovial fluids aspirated from saline injected controls. The results indicate that the intensity of visible fluorescence of equine synovial fluid increases after exposure to free radicals and during synovitis in the horse, suggesting a possible role for free radicals in the pathogenesis of equine inflammatory joint diseas