Detection of potato leaf roll virus and potato viruses M,S, X and Y by dot immunobinding on plain paper

Summary Potato leafroll virus and potato viruses M, S, X and Y in green leaves were detected by dot immunobinding (DIB) on plain paper and on nitrocellulose membranes. On both materials, DIB could detect the presence of very small amounts of virus, e.g. 30 pg of purified PVX. The sensitivity of the DIB test on plain paper and on nitrocellulose was compared to Double Antibody Sandwich ELISA (DAS-ELISA) by serial dilutions of infected plant sap made in healthy plant sap. Detection of potato viruses by DIB on plain paper and nitrocellulose was found to be equally sensitive whereas DAS-ELISA was 2 to 8 times more sensitive. Possible simplifications of the DIB procedure to suit the requirements of a routine method were examined. The use of the DIB method for routine testing of potato viruses is discussed.     

Development of inosine monophosphate and its degradation products during aging of pork of different qualities in relation to basic taste and retronasal flavor perception of the meat

Inosine monophosphate (IMP) and its degradation products, ribose and hypoxanthine, are all considered to be important constituents in meat flavor formation and development. The present study explored the fate of IMP during the aging of two qualities of pork (pH >5.7 and 5.5 < pH < 5.6) and the potential relationship between IMP, hypoxanthine, and sensory attributes of pork registered both as retronasal and basic taste responses in whole meat, meat juice, and the remaining meat residue. During aging the concentration of IMP decreased with a simultaneous increase in the concentrations of inosine, hypoxanthine, and ribose. The rates at which IMP was degraded to inosine and inosine to hypoxanthine during aging were found to be in agreement with the known rate constants of the dephosphorylation of IMP and the hydrolysis of inosine, respectively. Moreover, high-pH pork resulted in a significantly higher concentration of hypoxanthine throughout storage compared with low-pH pork due to an initially higher concentration of IMP in high-pH meat. The sensory analysis showed increasing intensity in bitterness and saltiness of pork as a function of aging, with the intensity being most pronounced in the meat juice. The increasing bitterness of the pork as a function of aging coincided with the higher content of hypoxanthine in these samples, thereby suggesting that degradation of IMP to hypoxanthine might influence pork flavor. In contrast, IMP was associated with nonaged meat and the sensory attributes meaty and brothy.     

Evaluation of skin erythema by use of chromametry and image analysis of digital photographs after intradermal administration of histamine in dogs

OBJECTIVE: To investigate whether the degree of erythema during an induced erythematous reaction, the histamine skin test reaction, can be assessed objectively by use of chromametry and image analysis of digital photographs. ANIMALS: 9 pet dogs (6 Golden Retrievers and 3 yellow Labrador Retrievers). PROCEDURE: Histamine phosphate was injected intradermally, and erythema of the wheal reaction was evaluated during the hour that followed. This was done by use of clinical scores, chromametry, and image analysis of digital photographs. Method reproducibility was tested for visual evaluation of printouts of digital photographs and for image analysis of the same photographs. RESULTS: The coefficient of variation of the technically derived erythema values was < 10%. The reproducibility of image analysis was high and the range of agreement between observers narrow. Using chromametry, it was not possible to differentiate between various degrees of erythema intensity as visually perceived. In contrast, use of image analysis of digital photographs enabled discrimination of slight erythema from moderate and marked erythema. The dynamics of reaction measured by chromametry followed the clinical observation. CONCLUSIONS AND CLINICAL RELEVANCE: Chromametric values are comparable to those obtained by visual inspection. As the result of standardized conditions, chromametry is preferred over digital photography.     

Handheld mechanical nociceptive threshold testing in dairy cows – intra‐individual variation,inter‐observer agreement and variation over time

ObjectiveTo examine the use of handheld methodology to assess mechanical nociceptive threshold (MNT) on cows kept loose-housed.Study designProspective randomized partial cross-over experimental study. A one-factor (test day) design was used to evaluate MNT over time.AnimalsOne hundred and fifteen healthy, loose-housed Danish Holstein cattle.MethodsWe evaluated intra-individual variation, inter-observer agreement and variation over time of MNT using two handheld devices and two stimulation sites. Mechanical, ramped stimulations were performed with an algometer (6.5 mm diameter steel probe, 0–10.0 kgf) or an electronic von Frey device (plastic tip with diameter 0.8 mm, 0–1000 gf). Each cow received 5–6 consecutive stimulations within a 2 × 5 cm skin area on the dorsal or lateral aspect of the left third metatarsus until an avoidance reaction occurred. We investigated the difference in precision [expressed as coefficient of variation (CV)] between the combinations of devices and stimulation sites. The inter-observer agreement and the difference in MNT between test day 1, 3, 7, 10 and 24 were investigated for selected combinations. Data were analysed in mixed models and Bland-Altman as relevant.ResultsThe CVs did not differ [range 0.34–0.52 (p = 0.1)]. Difference between observers (95% limits) was 0.2 kgf (2.8) and 4 gf (369) for the algometer and von Frey device, respectively. Mechanical nociceptive threshold increased from 361 on test day one to 495 gf on test day 24 (p < 0.01).Conclusion and clinical relevanceAll methods showed a high degree of intra-individual variation, and no combination of device and stimulation site showed superior precision. Mean difference between observers was low, and MNT was not consistent over time. Further development of the methods is required before they can be used in research to investigate possible relations between claw lesions and hyperalgesia.     

Histopathological nerve and skeletal muscle changes in rats subjected to persistent insulin-induced hypoglycemia

New insulin analogues with a longer duration of action and a flatter pharmacodynamic profile are developed to improve convenience and safety for diabetic patients. During the nonclinical development of such analogues, safety studies must be conducted in nondiabetic rats, which consequently are rendered chronically hypoglycemic. A rat comparator model using human insulin would be valuable, as it would enable differentiation between effects related to either persistent insulin-induced hypoglycemia (IIH) or a new analogue per se. Such a model could alleviate the need for an in-study-comparator and thereby reduce the number of animals used during development. Thus, the aims of the present study were i) to develop a preclinical animal model of persistent hypoglycemia in rats using human insulin infusion for four weeks and ii) to investigate histopathological changes in sciatic nerves and quadriceps femoris muscle tissue, as little is known about the response to persistent hypoglycemia in these tissues. Histopathologic changes in insulin-infused animals included axonal degeneration and myofibre degeneration. To our knowledge, this is the first study to show that persistent IIH provokes peripheral nerve and skeletal myofiber degeneration within the same animals. This suggests that the model can serve as a nonclinical comparator model during development of long-acting insulin analogues.     

Infrared skin temperature measurements for monitoring health in pigs: a review

Infrared temperature measurement equipment (IRTME) is gaining popularity as a diagnostic tool for evaluating human and animal health. It has the prospect of reducing subject stress and disease spread by being implemented as an automatic surveillance system and by a quick assessment of skin temperatures without need for restraint or contact. This review evaluates studies and applications where IRTME has been used on pigs. These include investigations of relationships between skin, ambient and body temperatures and applications for detecting fever, inflammation, lesions, ovulation, and stress as well as for meat quality assessment. The best skin locations for high correlation between skin temperature and rectal temperature are most likely thermal windows such as ear base, eye region and udder. However, this may change with age, stressors, and biological state changes, for example, farrowing. The studies performed on pigs using IRTME have presented somewhat discrepant results, which could be caused by inadequate equipment, varying knowledge about reliable equipment operation, and site-specific factors not included in the assessment. Future focus areas in the field of IRTME are suggested for further development of new application areas and increased diagnostic value in the porcine and animal setting in general.     

Use of chromametry and digital photography for objective measurement of skin color in clinically normal dogs

OBJECTIVE: To evaluate whether skin erythema in clinically normal dogs can be quantified by use of chromametry and image analysis of digital photographs. ANIMALS: 9 German Shepherd Dogs and 10 mixed-breed dogs. PROCEDURE: Hair was clipped at 7 sites on the body. Skin erythema was evaluated at the axillary region, right and left lateral aspect of thorax, right and left loin area (ie, part of the back between the thorax and pelvis), right and left groin area (ie, the junctional region between the abdomen and thigh), metatarsal digital pad, and on the nose. Replicate measurements were done by use of chromametry and image analysis of digital photographs, using erythema values in accordance with the Committee International d'Eclairage (CIE)-Lab color system. RESULTS: Repeatability was high for both techniques. Within-dog variation was lower than between-dog variation. Between-dog variation was high for both groups of dogs. Interregional variation was significant in German Shepherd Dogs and mixed-breed dogs. Erythema values revealed symmetry between the right and left lateral aspects of the thorax and loin and groin areas. CONCLUSIONS AND CLINICAL RELEVANCE: Precise objective methods are available for skin erythema quantification. Chromametric and photographic erythema values had a high within-dog reproducibility. Between-dog variability was high for German Shepherd Dogs and mixed-breed dogs as was regional variation, indicating differences in color among dogs.     

Two types of radicals in whole milk powder. Effect of lactose crystallization, lipid oxidation, and browning reactions

Whole milk powder was stored in closed vials at 60 degrees C to induce crystallization of lactose within a short time scale. After an induction period of 3-4 days simultaneous crystallization of lactose, increase of water activity, formation of browning products, and increase of radical content took place. Radicals detected before lactose crystallization were characterized by a narrow ESR spectrum (g = 2.006) and could be depleted by removal of oxygen and therefore were assigned to oxidation processes. Late-stage radicals present after crystallization of lactose gave much wider spectra (g = 2.0048) and were independent of oxygen availability and assigned to late-stage Maillard reaction products. The study indicates that the processes of lactose crystallization, browning, and formation of radical species (g = 2.0048) are strongly coupled, while lipid oxidation is less dependent on the other processes.