BKCa-Cav channel complexes mediate rapid and localized Ca2+-activated K+ signaling

Large-conductance calcium- and voltage-activated potassium channels (BKCa) are dually activated by membrane depolarization and elevation of cytosolic calcium ions (Ca2+). Under normal cellular conditions, BKCa channel activation requires Ca2+ concentrations that typically occur in close proximity to Ca2+ sources. We show that BKCa channels affinity-purified from rat brain are assembled into macromolecular complexes with the voltage-gated calcium channels Cav1.2 (L-type), Cav2.1 (P/Q-type), and Cav2.2 (N-type). Heterologously expressed BKCa-Cav complexes reconstitute a functional "Ca2+ nanodomain" where Ca2+ influx through the Cav channel activates BKCa in the physiological voltage range with submillisecond kinetics. Complex formation with distinct Cav channels enables BKCa-mediated membrane hyperpolarization that controls neuronal firing pattern and release of hormones and transmitters in the central nervous system.     

Improving the sensitivity of the IBR-gE ELISA for testing IBR marker vaccinated cows from bulk milk

The low sensitivity of the IBR-gE ELISA compared to other diagnostic ELISA tests for IBR is a major disadvantage of IBR control programmes based on IBR marker vaccination. Therefore the IBR-gE ELISA is not generally recommended for testing pooled or bulk milk samples.The aim of this study was to determine the performance of a commercially available kit for concentrating and purifying antibodies in milk in order to improve the sensitivity of detecting IBR-gE antibody positive cows from pooled and bulk milk samples. A single IBR-gE positive cow is likely to remain undetected in a pool of 49 negative milk samples without concentration. By contrast, the bulk milk concentration procedure improved sensitivity from 5.4% to 75.7% in a positive herd. Milk samples with a high or moderate positive signal are more likely to be detected after pool milk concentration compared to weak positive samples. Whereas a follow up study involving a monthly testing of bulk milk samples from three marker vaccinated IBR-gE negative herds over a period of seven months yielded negative results each month, bulk milk from a herd containing <5% IBR-gE positive cows always detected positive after concentration. Although the milk concentration procedure had no impact on specificity, it significantly enhanced the sensitivity of the detection of IBR-gE positive milk in pooled and bulk milk samples. After further evaluation this procedure could allow a cost efficient and reliable method of monitoring IBR marker-vaccinated herds for IBR-gE antibodies.     

Sweet blue lupin (Lupinus angustifolius L.) seed as a substitute for concentrate mix supplement in the diets of yearling washera rams fed on natural pasture hay as basal diet in Ethiopia

In the mixed crop-livestock farming system of Ethiopia where crop residues are the major feed resources and concentrate supplement feeds are not common, home-grown legume protein sources can help to minimise the feed problem. A 69-day feeding experiment on sheep was conducted to evaluate the potential of sweet blue lupin (Lupinus angustifolius L.) cultivar Sanabor seed as a substitute for commercial concentrate supplement. Thirty yearling male intact Washera sheep with initial body weight of 21 ± 1.38 kg (mean ± SD) were used. The design was a randomised complete block design with six replications. The five experimental supplement feeds were 453 g concentrate (T1), 342 g concentrate + 74 g lupin seed (T2), 228 g concentrate + 147 g lupin seed (T3), 116 g concentrate + 219 g lupin seed (T4) and 290 g lupin seed (T5) in dry matter basis to supplement around 100 g crude protein per day per animal. There were significant differences (P < 0.05) in total dry matter, crude protein, ash and organic matter intakes among treatments. The average daily body weight gain for T1, T2, T3, T4 and T5 was 91, 79, 79, 87 and 74 g/day, respectively, and this difference was not significant (P > 0.05). It was concluded that blue lupin seed has a potential to substitute the commercial concentrate supplement feed in Ethiopia.     

Establishment of diagnostic criteria for feline nonflea-induced hypersensitivity dermatitis

Hypersensitivity dermatitides (HD) are commonly seen in cats, and they are usually caused by environmental, food and/or flea allergens. Affected cats normally present with one of the following clinical reaction patterns: head and neck excoriations, usually symmetrical self-induced alopecia, eosinophilic skin lesions or miliary dermatitis. Importantly, none of these clinical presentations is considered to be pathognomonic for HD skin diseases, and the diagnosis of HD is usually based on the exclusion of other pruritic diseases and on a positive response to therapy. The objectives of this study were to propose sets of criteria for the diagnosis of nonflea-induced HD (NFHD). We recruited 501 cats with pruritus and skin lesions and compared clinical parameters between cats with NFHD (encompassing those with nonflea, nonfood HD and those with food HD), flea HD and other pruritic conditions. Using simulated annealing techniques, we established two sets of proposed criteria for the following two different clinical situations: (i) the diagnosis of NFHD in a population of pruritic cats; and (ii) the diagnosis of NFHD after exclusion of cats with flea HD. These criteria sets were associated with good sensitivity and specificity and may be useful for homogeneity of enrolment in clinical trials and to evaluate the probability of diagnosis of NFHD in clinical practice. Finally, these criteria were not useful to differentiate cats with NFHD from those with food HD.     

Digital Agenesia in Martina Franca Donkey Foal: A Case Report

A 4-day-old male Martina Franca donkey foal was evaluated for a forelimb alteration. Clinical examination and radiographs revealed the agenesia of the distal digit. Biochemical parameters were normal, and ultrasonographic evaluation did not identify any relievable organ alteration. Karyotype study revealed an abnormality on chromosome 1. The foal was discharged with a distal limb bandage in which a palmar splint was applied. A poor prognosis for the functionality of the limb was given. In endangered species, such as the Martina Franca donkey, the excessive inbreeding could result in an increase in genetic disorders. These findings shed new light on the possible pathogenesis of the digital dysgenesia. The study of the karyotype could be a useful approach to detect genetic alterations that could or could not be expressed in the animal, especially in endangered species in which a risk of an excessive inbreeding is considerable. These defects should be considered in the choice and selection of the breeders.     

In vitro antifungal susceptibility of Malassezia pachydermatis from dogs with and without skin lesions

Canine Malassezia dermatitis is frequently treated with systemic ketoconazole (KTZ) and itraconazole (ITZ). However, no information is available on the antifungal susceptibility to azoles and allilamine of Malassezia pachydermatis isolates from dogs with or without skin lesions. The present study was designed to evaluate the in vitro antifungal susceptibility of M. pachydermatis strains from dogs with or without skin lesions to KTZ, ITZ, miconazole (MICO), fluconazole (FLZ), posaconazole (POS), voriconazole (VOR) and terbinafine (TER) using the Clinical and Laboratory Standards Institute reference Broth Microdilution Method (CLSI M27-A2). The association between the susceptibility to antifungal compounds and the origin of M. pachydermatis, from skin with or without lesions has been also assessed. A total of 62 M. pachydermatis strains from healthy dogs (i.e., Group A=30) or with skin lesions (i.e., Group B=32) were tested. ITZ, KTZ and POS showed the highest activity against M. pachydermatis strains, whereas MICO TER and FLZ the lowest. A higher number of Malassezia resistant strains were registered among isolates from Group B than those from Group A. This study indicates that M. pachydermatis strains were susceptible to ITZ, KTZ, and POS. However, dogs with lesions may harbour strains with low susceptibility to antifungal agents and displaying cross-resistance phenomena to azole. The antifungal therapy in Malassezia infections requires careful appraisal of choice of drugs especially in cases of unresponsiveness to antifungal treatment or recurrent infections.     

Influence of time of food administration on daily rhythm of total locomotor activity in ponies

The activity rhythm can be entrained by nonphotic cues, especially food availability. Daily feeding schedules can act as “Zeitgeber” to synchronize the circadian system. In this study, the authors evaluated the influence of different feeding schedules on the daily rhythm of total locomotor activity in ponies. Six clinically healthy Shetland ponies were kept under natural 12/12-h light/dark cycle and monitored during 4 different feeding schedules: fed twice (at 8 am and 8 pm), fasted, fed at 8 pm, fed at 8 am, and fed twice a day again. Water was available ad libitum. To record total activity, we equipped the animals with actigraphy-based data loggers (Actiwatch-Mini). Two-way analysis of variance revealed a significant main effect of the time of day and no effect of feeding schedules on the amount of total locomotor activity. Locomotor activity showed daily rhythmicity in all experimental conditions, with diurnal acrophase. The amplitude of the rhythms and their robustness statistically changed across all experimental conditions. Our findings support the theory that when given essentially free choice hay, the time of feeding did not entrain activity, and suggest that feeding behavior can influence the distribution of locomotor activity during a 24-hour period in ponies, with changes in the amplitude and robustness of the rhythm.