Imerslund-Grasbeck syndrome in a cross-breed dog

Imerslund-Gräsbeck syndrome is an autosomal recessive disease reported only in certain pure-breed dogs. An 18-month-old, male neutered beagle cross-breed was presented for evaluation of severe lethargy, progressive weakness and anorexia. Main clinicopathological findings included low body condition score (2.5/9), severe muscle atrophy, several neurological abnormalities, mild normochromic, normocytic, non-regenerative anaemia, severe hypocobalaminemia and mild proteinuria. Extensive diagnostic tests ruled out most of differential diagnoses for the aforementioned clinicopathological abnormalities and genetic evaluation showed that the dog was heterozygous for two previously described mutations affecting the CUBN gene, the beagle and the border collie variants. The dog showed an excellent clinical response to oral cobalamin supplementation with no relapse after 4 months. In conclusion, this case creates awareness that Imerslund-Gräsbeck syndrome should be considered even in mixed-breed dogs with compatible clinical signs and that two different pathogenic CUBN mutations in compound heterozygosity can lead to a typical Imerslund-Gräsbeck syndrome phenotype.     

Night-time Melatonin Secretion and Seasonally Delayed Puberty in Gilts

This study was conducted to determine whether the seasonal delay in puberty in autumn is driven by individual differences in night-time melatonin secretion in domestic gilts at the attainment of puberty. A group of spring-born gilts (n = 30) were expected to reach puberty in autumn by the age of 7 months. Eighteen of these gilts were selected in pairs on the basis of matched days of birth. By the expected time, half of the animals showed oestrous symptoms (group CYCLING, n = 9) with the rest remaining silent (group SILENT, n = 9). Afterwards, all gilts were fitted with indwelling jugular catheters for frequent blood sampling. Blood samples were collected from all animals three times during the day followed by three times in the night at 2-h intervals for 48 h. The samples were analysed by a commercial radioimmunoassay (RIA). The results show a consistent 25-fold rise (on average) in night-time melatonin concentration in every animal sampled with group averages ranging from 0.28 +/- 0.04 to 0.37 +/- 0.06 pg/ml at day and from 10.20 +/- 2.16 to 10.67 +/- 0.05 pg/ml at night. Night-time group mean values between CYCLING and SILENT gilts did not differ significantly (10.26 +/- 0.67 and 10.38 +/- 0.94 for the CYCLING; 10.67 +/- 0.05 and 10.20 +/- 2.16 for the SILENT). When 10 pg/ml was used as a threshold value, six individuals did not reach it during the night (low responders). Two of these gilts were CYCLING and four were SILENT. In conclusion, the results presented imply no involvement of the level of night-time melatonin concentration in the seasonal delay of puberty in gilts.     

Factors Affecting Fertility in Loosely Housed Sows and Gilts: Vulvar Discharge Syndrome, Environment and Acute-phase Proteins

The effect of vulvar discharge syndrome (VDS) on sow and gilt fertility was studied on 26 farms. Of 824 animals inspected in 21 randomly selected and five VDS problem farms, 19 (2.3%) were afflicted with VDS. Altogether 542/799 of the examined animals (67.8%) farrowed thereafter. Nine of the 19 VDS animals (47.4%) and 533/780 non-VDS animals (68.3%) farrowed at the first chance after the examination (p = 0.05). None of the unmated gilts in this study had VDS. Environmental and individual factors likely to be associated with fertility and VDS were tested. In multivariate analysis, factors associated with farrowing were VDS, reproductive status, availability of roughage and confinement to individual stalls. None of the variables tested was associated with VDS. However, all of the VDS problem farms were overcrowded and had concrete, partly slatted floors with little or no bedding. The median value of haptoglobin (Hp) was 2.5 (range 1.3-3.1) g/l in VDS animals and 2.3 (0.5-4.3) g/l in controls (p = 0.6). The median C-reactive protein (CRP) in VDS animals was 30.3 (3.3-171.3) mg/l and in controls 25.9 (3.3-361.1) mg/l (p = 0.7). In conclusion, VDS decreased fertility of gilts and sows in the absence of a systemic acute-phase response, as indicated by stable concentrations of Hp and CRP.     

Evaluation in mice of Brucella ovis attenuated mutants for use as live vaccines against B. ovis infection

Brucella ovis causes ram contagious epididymitis, a disease for which a specific vaccine is lacking. Attenuated Brucella melitensis Rev 1, used as vaccine against ovine and caprine brucellosis caused by B. melitensis, is also considered the best vaccine available for the prophylaxis of B. ovis infection, but its use for this purpose has serious drawbacks. In this work, two previously characterized B. ovis attenuated mutants (Δomp25d and Δomp22) were evaluated in mice, in comparison with B. melitensis Rev 1, as vaccines against B. ovis. Similarities, but also significant differences, were found regarding the immune response induced by the three vaccines. Mice vaccinated with the B. ovis mutants developed anti-B. ovis antibodies in serum of the IgG₁, IgG_2a and IgG_2b subclasses and their levels were higher than those observed in Rev 1-vaccinated mice. After an antigen stimulus with B. ovis cells, splenocytes obtained from all vaccinated mice secreted similar levels of TNF-α and IL12(p40) and remarkably high amounts of IFN-γ, a crucial cytokine in protective immunity against other Brucella species. By contrast, IL-1α -an enhancer of T cell responses to antigen- was present at higher levels in mice vaccinated with the B. ovis mutants, while IL-10, an anti-inflammatory cytokine, was significantly more abundant in Rev 1-vaccinated mice. Additionally, the B. ovis mutants showed appropriate persistence, limited splenomegaly and protective efficacy against B. ovis similar to that observed with B. melitensis Rev 1. These characteristics encourage their evaluation in the natural host as homologous vaccines for the specific prophylaxis of B. ovis infection.     

Use of nootkatone as a senescence indicator for Rouge La Toma Cv. grapefruit (Citrus paradisi Macf.)

The objective of this research was to study the usefulness of nootkatone as a senescence indicator for Rouge La Toma cv. grapefruit (Citrus paradisi Macf.), simulating different treatments that included the normal postharvest handling of citrus fruits: temperature conditioning, cold storage, shipment periods to overseas markets such as Japan and the U.S., marketing conditions, and storage at nonchilling temperature (control treatments). The highest nootkatone levels, determined by GLC-MS analyses, were detected in fruits subjected to control treatments. No significant differences were observed in nootkatone levels between treatments either with or without temperature conditioning prior to the start of the cold storage. Levels of nootkatone increased throughout time for all assayed treatments. The linear regressions of nootkatone levels showed correlation coefficients of 0.80 and 0.83 with storage time (29 and 42 days, respectively). Therefore, nootkatone appears to be a good indicator of senescence for Rouge La Toma grapefruit.     

Determination of thiamin in cooked sausages

A reliable and rapid high-performance liquid chromatography (HPLC) method has been set up for the determination of total thiamin in difficult sample matrices such as cooked sausages. Different hydrolysis conditions and enzymes were tested to release the vitamin from its phosphate ester. The best data in the enzymatic digestion were obtained by incubating the samples with 6% clara-diastase at 50 degrees C for 3 h. After oxidation of thiamin to thiochrome, the sample extracts were purified by using a C(18) Sep-Pak cartridge. Final determination was performed by reversed-phase HPLC with fluorescence detector (excitation 360 nm, emission 430 nm), on a low-cost 25 cm x 4 mm i.d. Spherisorb C(8) cartridge using a mixture of 5 mM phosphate buffer pH 7.0 and acetonitrile (70:30, v/v) as mobile phase. Precision of the method was 1.5% (within a day) and 5. 2% (between days). The detection limit was 0.015 mg/100 g. All the recoveries from the different cooked sausages were better than 90% of thiamin hydrochloride added to samples of meats. In the samples analyzed, the mean value for thiamin was between 0.039 and 0.508 mg/100 g fresh weight.     

Geomorphological significance of lichen colonization in a present snow hollow: Hoya del Cuchillar de las Navajas, Sierra de Gredos (Spain)

This paper discusses the results of a lichenometrical and geomorphological study of one of the few remaining active snow hollows in the central region of the Iberian Peninsula. The study area, located on a glacial shoulder, is called Hoya del Cuchillar de las Navajas. A protalus rampart occurs at the base of the hollow. Our studies, conducted between 1992 and 1998, were designed to determine the geomorphological characteristics of Hoya, the mobility of the deposits, and the characteristics of the snow cover. These data formed the basis for a study of the lichen colonization on the blocks and on the wall surrounding the snow hollow. All of the lichen species found were analyzed according to their abundance, distribution and the extent of their surface cover. Measurements of the diameter of the thalli of the species Rhizocarpon geographicum were also obtained. Thalli of this species were found to require a mean snow-free growing season of at least 95 days (13.5 weeks) per year. Maximum mean thallus diameters indicate that the protalus rampart was formed during the Little Ice Age and became inactive 130 years ago.     

Evaluation of auditory evoked potentials to predict depth of anaesthesia during fentanyl/fluanisone−midazolam anaesthesia in rats

Objectives To assess a method for monitoring depth of anaesthesia using components of middle latency auditory evoked potential (AEP) waveforms during anaesthesia with fentanyl/fluanisone and midazolam. Study design Prospective observational study. Animals Five female Wistar rats weighing between 210 and 250 g. Methods Implanted electrodes were used to record AEPs in animals receiving five doses of anaesthetic. Recordings were made at 5 minutes post‐injection (deep anaesthesia; no pedal withdrawal response, PWR) and then at 25 minutes (light anaesthesia; strong PWR). Responses showed five characteristic peaks occurring at 11, 14, 23, 42 and 68 ms that were measured for latency of occurrence and peak amplitude. Results Auditory evoked potential peaks P14, N23 and P42 were increased significantly in latency with successive anaesthetic injections [avg. F(1,4) = 12.53, p < 0.001; avg. F(1,4) = 10.6, p < 0.001; avg. F(1,4) = 3.9, p = 0.02, respectively]. Peak N23 showed a significant reduction in latency during the 20 minute recovery period following both the first and second anaesthetic injections (t(3) = 7.52, p = 0.005; t(4) = 5.17, p = 0.007, respectively). Peak P42 occurred significantly earlier 20 minutes following the second anaesthetic injection (t(4) = 4.75, p = 0.009). The mean overall depth of anaesthesia assessed using PWR scores was significantly correlated with the mean latency of peak N23, such that as the strength of PWR increased, N23 occurred significantly earlier (r = ?0.99, p = 0.01). The amplitude difference between peaks N23 and P42 increased after the second and third drug administrations [avg. F(1,4) = 10.65, p = 0.031 and avg. F(1,4) = 11.24, p = 0.028, respectively]. Conclusion The characteristics of these peaks, and in particular latency of peak N23, may provide a useful tool for assessing depth of anaesthesia produced by this, and possibly other anaesthetic agents.