A quantitative approach to analysing cortisol response in the horse

The cortisol response to glucocorticoid intervention has, in spite of several studies in horses, not been fully characterized with regard to the determinants of onset, intensity and duration of response. Therefore, dexamethasone and cortisol response data were collected in a study applying a constant rate infusion regimen of dexamethasone (0.17, 1.7 and 17 μg/kg) to six Standardbreds. Plasma was analysed for dexamethasone and cortisol concentrations using UHPLC‐MS/MS. Dexamethasone displayed linear kinetics within the concentration range studied. A turnover model of oscillatory behaviour accurately mimicked cortisol data. The mean baseline concentration range was 34–57 μg/L, the fractional turnover rate 0.47–1.5 1/h, the amplitude parameter 6.8–24 μg/L, the maximum inhibitory capacity 0.77–0.97, the drug potency 6–65 ng/L and the sigmoidicity factor 0.7–30. This analysis provided a better understanding of the time course of the cortisol response in horses. This includes baseline variability within and between horses and determinants of the equilibrium concentration–response relationship. The analysis also challenged a protocol for a dexamethasone suppression test design and indicated future improvement to increase the predictability of the test.     

Uterine immune reaction and reproductive performance of sows inseminated with extended semen and infused with pooled whole dead semen

The objective of this study was to investigate the effect of infusing whole dead semen (WDS) after AI with diluted commercial semen on uterine inflammatory reaction and embryonic survival rate in gilts. Sixty Yorkshire-Landrace gilts were assigned at their second estrus to one of the following AI treatments: 1) commercial semen adjusted to 1 x 10(9) sperm cells (S1) per dose, followed by an infusion of 80 mL of WDS (S1-WDS); 2) S1 followed by an infusion of 80 mL of Beltsville Thawing Solution (S1-BTS); 3) commercial semen adjusted to 3 x 10(9) sperm cells (S3) per dose, followed by an infusion of 80 mL of BTS (S3-BTS); and 4) a negative control group, in which gilts received two infusions of 80 mL of BTS (BTS). Two days after the first AI, eight gilts from Groups 1, 2, and 4 were slaughtered and reproductive tracts were collected. One horn was cut open longitudinally along the antimesometrial aspect and endometrial samples were taken and immediately frozen for analysis of messenger RNA (mRNA) abundance for inflammatory cytokines and growth factors. The other horn was flushed with 20 mL of PBS, and the contents of interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) were determined by ELISA. On d 25 after AI, gilts from Groups 1, 2, and 3 were slaughtered and their reproductive tracts were collected to evaluate the number of fetuses and corpora lutea. On d 2 after the first AI, only TGF-beta1 was detected in the flush of all gilts, and no difference was observed between S1-WDS, S1-BTS, and BTS gilts. Endometrial levels of IFN-gamma and interleukin (IL)-6 mRNA were marked in all gilts, but they were not affected by the AI treatments, whereas the mRNA abundances for IL-1 and IL-2 were negligible. Infusions of WDS or BTS after a fertile AI did not affect IGF-I, IGF-I receptor, or IGF-II mRNA levels compared with gilts infused with BTS only, whereas the mRNA abundance for the IGF-II receptor was decreased (P < 0.05) in WDS-infused gilts. In gilts inseminated with S1 doses, infusion of WDS did not affect the number of live embryos. Although infusions of WDS did not affect the mRNA level and secretion of the cytokines measured and did not improve embryonic survival rates, further studies are needed to better understand the influence of semen composition on the uterine response after mating.     

Feasibility of using rare earth elements (REEs) to mark and identify escaped farmed Atlantic salmon Salmo salar L.

A series of rare earth elements (REEs) were evaluated as potential markers in scales of Atlantic salmon 1⁺ and 0⁺ smolt by addition of these elements to the feed. Chlorides of 5 REEs were tested, alone or in combination: lanthanum (La), cerium (Ce), praseodymium (Pr), neodymium (Nd) and dysprosium (Dy). The labelled‐diets contained 125 or 250 mg REE kg^?1 feed and were administered for up to 10 weeks. Fish scales were collected and analysed by inductively coupled plasma‐mass spectrometry (ICP‐MS) before start, 5 and 10 weeks after labelling started and finally 2 or 4 months after marking. The results demonstrated that the five tested elements were clearly incorporated into the scales of the fish fed the supplemented diets. Uptake and concentrations increased gradually during the feeding period, and no principal differences were found between the two smolt types. Combining 2 REEs in the same feed did not affect the incorporation of either of them. After a ‘dilution’‐period of 2–4 months the concentrations of all markers (except La) in the scales of the treated‐fish were still significantly higher than those of the untreated‐fish. The background concentrations of the tested markers and some other elements were also measured and found to give interesting information.     

Molecular characterization of the incitant of cowpea bacterial blight and pustule, Xanthomonas campestris pv. vignicola

Strains of Xanthomonas campestris pv. vignicola (Xcv), isolated from cowpea leaves with blight or minute pustules and collected from various geographic areas, were selected on the basis of pathological and physiological features. All strains were analyzed for genotypic markers by two methods: ribotyping with EcoRI endonuclease, and RFLP analysis with a plasmid probe (pthB) containing a gene required for pathogenicity from Xanthomonas campestris pv. manihotis. Ribotyping revealed a unique pattern for all the strains that corresponded to the previously described ribotype rRNA7. Based on polymorphism detected by pthB among Xcv strains, nine haplotypes were defined. The observed genetic variation was independent of the geographic origin of the strains and of pathogenic variation. Some haplotypes were widely distributed, whereas others were localized. In some cases, we could differentiate strains isolated from blight symptoms and pustules according to haplotypic composition. However, in most cases, no significant differences were observed. Our results and the previous pathogenic and biochemical characterizations suggest that the strains isolated from leaves with blight symptoms or minute pustules belong to the same pathovar. We provide information on pathogen diversity that can be used to identify and characterize resistant germplasm.     

Ketoprofen reduces the minimum alveolar concentration (MAC) in dogs anaesthetized with isoflurane and fentanyl

Non‐steroidal anti‐inflammatory drugs may potentiate the opioid induced reduction in volatile anaesthetic requirements ( Gomez de Segura et al. 1998 ). This study determined the reduction in the MAC of isoflurane (ISO) produced by ketoprofen (KETO) in dogs anaesthetized with fentanyl (FENT) and ISO. Six healthy female crossbred dogs, weighing 13.5 ± 1.3 (mean ± SD) kg and aged 3.0 ± 0.9 years were studied. Approval of the study was obtained from the institutional ethics committee. Anaesthesia was induced in all dogs via a facemask with 5% ISO in 5 L minute^?1 oxygen. The dogs' trachea were intubated and lungs were ventilated to maintain normocapnia (Pe ′CO₂ 4.7–6 kPa, 35–45 mm Hg). A heating pad was used to maintain body temperature. The animals were anaesthetized four times at one week intervals with the following anaesthetic and analgesic protocols randomly administered. Study 1, MAC (ISO); Isoflurane MAC. Study 2, MAC (ISO + FENT); dogs anaesthetized with ISO received a loading dose of 30 µg kg^?1 FENT IV over 20 minutes followed by a maintenance infusion of 0.2 µg kg^?1 minute^?1 FENT. Study 3, MAC (ISO + FENT + KETO1); as study 2 plus 1 mg kg^?1 KETO. Study 4, MAC (ISO + FENT + KETO2); as study 2 plus 2 mg kg^?1 KETO. The MAC was determined in duplicate by applying a standard electrical stimulus (50 V, 50 H₂ over 60 seconds via two needles placed SC over the tarsus). The stimulus was applied 15 minutes after every step change in anesthetic concentration. The Wilcoxon test was applied to data to determine significant differences among MAC measurements. Fentanyl significantly decreased MAC (ISO) from 1.27% ± 0.02% to 0.73% ± 0.08%, a reduction of 42% (p < 0.05). Ketoprofen 1 mg kg^?1 further decreased the MAC value (although not statistically significantly) with a reduction of 47% from MAC (ISO) (0.67% ± 0.13%) and 8% from MAC (ISO + FENT). When KETO 2 mg kg^?1 was given, the reduction in MAC was 50% compared to MAC (ISO) (0.63% ± 0.08%; p < 0.05) and 14% compared to MAC (ISO + FENT) p < 0.05. Administration of KETO further reduces MAC (ISO) compared to levels observed with FENT alone. The observed reduction may have clinical advantages.     

Are the number and size of scales in Liolaemus lizards driven by climate?

Ectothermic vertebrates are sensitive to thermal fluctuations in the environments where they occur. To buffer these fluctuations, ectotherms use different strategies, including the integument, which is a barrier that minimizes temperature exchange between the inner body and the surrounding air. In lizards, this barrier is constituted by keratinized scales of variable size, shape and texture, and its main function is protection, water loss avoidance and thermoregulation. The size of scales in lizards has been proposed to vary in relation to climatic gradients; however, it has also been observed that in some groups of Iguanian lizards it could be related to phylogeny. Thus, here, we studied the area and number of scales (dorsal and ventral) of 60 species of Liolaemus lizards distributed in a broad latitudinal and altitudinal gradient to determine the nature of the variation of the scales with climate, and found that the number and size of scales are related to climatic variables, such as temperature and geographical variables as altitude. The evolutionary process that best explained how these morphological variables evolved was the Ornstein‐Uhlenbeck model. The number of scales seemed to be related to common ancestry, whereas dorsal and ventral scale areas seemed to vary as a consequence of ecological traits. In fact, the ventral area is less exposed to climate conditions such as ultraviolet radiation or wind and is, thus, under less pressure to change in response to alterations in external conditions. It is possible that scale ornamentation, such as keels and granulosity, may bring some more information in this regard.     

Experimental Neospora Caninum Infection in Pregnant Dairy Heifers Raises Concentrations of Pregnancy‐Associated Glycoproteins 1 and 2 in Foetal Fluids

Plasma concentrations of PAG‐1 are used for pregnancy diagnosis and as a marker of placental/foetal well‐being, while those of PAG‐2 may be an indicator of abortion risk in Neospora caninum‐infected cows. Studies have shown that N. caninum infection modifies PAG‐1 and PAG‐2 patterns in maternal blood plasma. However, no prior work has examined the effects of N. caninum infection on concentrations of PAGs in foetal fluids. In this study, PAG‐1, PAG‐2 and pH levels were determined in the amniotic and allantoic fluids of foetuses collected at 152 days of gestation from control uninfected dams and from dams experimentally infected with N. caninum on Day 110 of gestation. Foetal fluids from infected foetuses had significantly higher PAG‐2 concentrations (p = 0.026) and pH values (p = 0.02) than fluids from non‐infected foetuses. In infected foetuses, significantly higher concentrations of PAG‐1 (p < 0.001) and PAG‐2 (p < 0.001) were detected in fluid samples showing antibodies against N. caninum than those without antibodies. Moreover, pH values were significantly higher (p = 0.011) in foetal fluid samples with antibodies than in samples from non‐infected foetuses. In conclusion, this is the first report on the effect of N. caninum infection on PAG levels in foetal fluids. Our results indicate that following the experimental infection of dams with N. caninum on Day 110 of gestation, foetal fluids collected from the infected foetuses of these dams featured higher PAG‐1 and PAG‐2 levels and pH values than fluids from non‐infected controls, provided that the samples tested showed the presence of antibodies. The clinical implications of these findings are that following infection with N. caninum, most cows will experience some level of placental damage and that this injury correlates with foetal fluid PAG levels and pH.