Evaluation of an in-clinic Serum Amyloid A (SAA) assay and assessment of the effects of storage on SAA samples

Background

An in-clinic assay for equine serum amyloid A (SAA) analysis, Equinostic EVA1, was evaluated for use in a clinical setting. Stability of SAA in serum samples was determined.

Methods

Intra- and inter- assay variation of the in-clinic method was determined. The in-clinic method (EVA1) results were compared to a reference method (Eiken LZ SAA) with 62 patient samples. For samples with SAA concentrations within the assay range of EVA1 (10-270 mg/L), differences between the methods were evaluated in a difference plot. Linearity under dilution was evaluated in two samples. Stability of SAA in three serum pools stored at 4°C and approximately 22°C was evaluated with the reference method day 0, 1, 2, 4, 7, 17 and analysed with a two-way ANOVA.

Results

The imprecision (coefficient of variation, CV) for the in-clinic method was acceptable at higher SAA concentrations with CV values of 7,3-12%, but poor at low SAA concentrations with CV values of 27% and 37% for intra- and inter-assay variation respectively. Recovery after dilution was 50-138%. The in-clinic assay and the reference method identified equally well horses with low (<10 mg/L) and high (>270 mg/L) SAA concentrations. Within the assay range of the in-clinic method, 10-270 mg/L, the difference between the two methods was slightly higher than could be explained by the inherent imprecision of the assays. There were no significant changes of serum SAA concentrations during storage.

Conclusions

The in-clinic assay identified horses with SAA concentrations of <10 mg/L and >270 mg/L in a similar way as the reference method, and provided an estimate of the SAA concentration in the range of 10-270 mg/L. The imprecision of the in-clinic method was acceptable at high SAA concentrations but not at low concentrations. Dilution of samples gave inconsistent results. SAA was stable both at room temperature and refrigerated, and thus samples may be stored before analysis with the reference method.     

Postprandial glycemia, insulinemia, and satiety responses in healthy subjects after whole grain rye bread made from different rye varieties. 1

Rye products typically induce low insulin responses and appear to facilitate glucose regulation. The objective of this study was to investigate differences in postprandial glucose, insulin, and satiety responses between breads made from five rye varieties. Breads made from whole grain rye (Amilo, Rekrut, Dankowski Zlote, Nikita, and Haute Loire Pop) or a white wheat bread (WWB) were tested in a randomized cross-over design in 14 healthy subjects (50 g available starch). Metabolic responses were also related to the composition of dietary fiber and bioactive compounds in the breads and to the rate of in vitro starch hydrolysis. The Amilo and Rekrut rye breads induced significantly lower insulin indices (II) than WWB. Low early postprandial glucose and insulin responses (tAUC 0-60 min) were related to higher amounts of caffeic, ferulic, sinapic, and vanillic acids in the rye breads, indicating that the phenolic acids in rye may influence glycemic regulation. All rye breads induced significantly higher subjective feelings of fullness compared to WWB. A low II was related to a higher feeling of fullness and a lower desire to eat in the late postprandial phase (180 min). The data indicate that some rye varieties may be more insulin-saving than others, possibly due to differences in dietary fiber, rate of starch hydrolysis, and bioactive components such as phenolic acids.     

Frequency of Salmonella detection in a broiler flock depending on different litter materials--a field study

During the fattening period of a broiler flock four different litter materials (peat, chopped straw, shavings, silage) were tested simultaneously.The separated sections were tested for the presence of Salmonella bacteria using the sock-sampling method as described in the regulation EC No. 646/2007 with slight modifications in the sampling technique and the laboratory protocol. In addition, some chemical and physical parameters regarding litter quality and house climate were analysed. Samples were taken at day 0 (housing of animals), day 14, and day 30. At the end of the fattening period Salmonella Paratyphi B (d-tartrate +), representing a common strain in German broiler flocks, were isolated from culture. According to the various types of bedding materials some differences in the frequency of Salmonella detection were found. With three samples out of six samples being positive, wood shavings showed the highest Salmonella prevalence, followed by peat with two positive results. Corn silage was found to be a suitable alternative for common litter materials and revealed only one positive sample. Chopped straw was found to be free of Salmonella in our study. However, the latter had a higher risk for negative side effects concerning animal health and house climate. The study showed that the choice of an appropriate litter material might be of considerable importance in order to decrease the Salmonella burden within poultry flocks.     

Detection of Mycobacterium avium ssp. paratuberculosis in ileocaecal lymph nodes collected from elderly slaughter cows using a semi-nested IS900 polymerase chain reaction

The aim of this study was to investigate the occurrence of subclinical Mycobacterium avium spp. paratuberculosis (MAP) infections at slaughter by testing ileocaecal lymph nodes with a semi-nested IS900 PCR. Tissue samples were available within the framework of a parallel study investigating BSE-susceptibility factors in members of BSE-cohorts in the German Federal State of Lower Saxony. Ileocaecal lymph nodes were collected over a 2-year sampling period from 99 slaughter cattle of a mean age of 6.5 years (5.5-7.5 years). A recently developed IS900 semi-nested polymerase chain reaction (snPCR) assay offering a sensitivity of 1 genome equivalent was used for the detection of MAP-DNA. Based on this snPCR, 17 out of the 99 samples gave positive results, indicating a MAP occurrence of 17.17% in the random sample. All PCR products were sequenced for screening of polymorphisms. Nucleotide homologies of 98.5-100% were found with respect to the MAP K10 reference sequence IS900 (GenBank: AE16958). PCR analysis of ileocaecal lymph nodes collected from slaughter cattle proved to be a suitable technique to determine MAP occurrence in the local cattle population.     

Subgroups of canine antinuclear antibodies in relation to laboratory and clinical findings in immune-mediated disease

Background:  Autoimmune system diseases in dogs are commonly referred to as systemic lupus erythematosus (SLE), with a positive antinuclear antibody (ANA) test as a hallmark. In human patients, other systemic ANA-positive diseases with overlapping diagnostic features, referred to as SLE-related diseases, are described.  Objectives:  The objective of this study was to investigate whether different patterns of ANA reactivity represent different systemic autoimmune diseases in dogs. Methods:  Dogs with serum positive for ANA by indirect immunofluorescence (IIF-ANA, titer ≥1:100) (n=56) were identified retrospectively from the patient population at the Department of Small Animal Clinical Sciences, Swedish University of Agricultural Sciences. Dogs were grouped on the basis of ANA staining patterns, and the results of immunodiffusion tests. Clinical, hematologic, serum biochemical, radiologic, and pathologic examinations were described for each group.  Results:  Dogs with a chromosomal–positive, homogeneous ANA staining pattern (n=14) had clinical signs involving multiple organ systems; 8 dogs were anemic. Dogs with a speckled IIF-ANA staining pattern (n=42) primarily had clinical signs of musculoskeletal disorders, fatigue and fever. Precipitating antibodies by immunodiffusion were found only in dogs with a speckled IIF-ANA staining pattern and comprised 4 different subgroups based on antigen specificity. Conclusions: In dogs with homogeneous IIF-ANA staining, SLE is a probable diagnosis because of the diversity of clinical manifestations and autoantibody reactivity against chromosomal antigens. Dogs with a speckled IIF-ANA pattern may have SLE-related diseases, which, in turn, may be correlated with different immunodiffusion subgroups. These syndromes had overlapping clinicopathologic features, as described for human patients.     

Gastrointestinal stromal tumour and hypoglycemia in a Fjord pony: Case report

Background

Neoplasia may cause hypoglycemia in different species including the horse, but hypoglycemia has not previously been reported in the horse associated with gastrointestinal stromal tumours.

Case presentation

A case of a gastrointestinal stromal tumour in a Fjord pony with severe recurrent hypoglycemia is presented. The mechanism causing the hypoglycemia was not established.

Conclusion

This case indicates that a gastrointestinal stromal tumour may cause hypoglycemia also in the horse.     

Quality of Filleted Atlantic Mackerel (Scomber Scombrus) During Chilled and Frozen Storage: Changes in Lipids,Vitamin D,Proteins, and Small Metabolites,including Biogenic Amines

Quality changes of vacuum-packed Atlantic mackerel (Scomber scombrus) fillets during 12 months’ frozen storage at ?27°C and 9 days’ chilled storage at +4°C were evaluated. Freezing at ?27°C preserved the long chain n-3 polyunsaturated fatty acids (LC n-3 PUFAs), both in light and dark muscle, vitamin D, and the low molecular weight metabolites (LMW) (studied by high resolution nuclear magnetic resonance spectroscopy, HR NMR). Protein oxidation took place, especially between 1 and 7 months, decreasing water holding capacity and protein extractability. During chilled storage, no lipid or protein oxidation was observed, but lipolysis increased, and several LMW metabolites relevant for sensory and nutritional quality degraded into non-favorable compounds. The content of biogenic amines was high at day 9 (e.g., 18 mg histamine/100 g), jeopardizing safety. Preservation of mackerel fillets by freezing at ?27°C is thus a better option compared to prolonged chilled storage at +4°C; the quality was well preserved for 12 months’ frozen storage.     

Organophosphate poisoning of Atlantic salmon in connection with treatment against salmon lice

Three incidents with high mortality in Atlantic salmon after trichlorfon treatment against salmon lice are described. All 3 incidents occurred at water temperatures of 12 degrees C or higher. The mean brain acetylcholinesterase (AChE) activity of dead fish was less than 20% of normal activity, while survivors showed mean activities of 22-61% of normal levels. Dichlorvos residues in muscular and liver tissues ranged from nondetectable levels to 0.2 micrograms/g tissue. The strongest inhibition of brain AChE was found in association with the highest dichlorvos residues. Substantial AChE-inhibition was, however, also found in samples in which dichlorvos residues could not be detected. AChE-determination was found to be more reliable than residue analysis for the diagnosis of organophosphate poisoning in salmon.     

Tryptophan Metabolism in Pigs: Relation to Synthesis and Function of Serotonin

Contrary to blood serotonin the level of free and total tryptophan is distinctly influenced by feed intensity and composition. Piglets show a considerably higher plasma free tryptophan and blood serotonin content than adult pigs, just as boars possess a significantly higher blood level of tryptophan and serotonin than sows. A pronounced individual variation of blood serotonin and a much less variable content among littermate pigs provide evidence that blood serotonin is genetically controlled. Tryptophan is readily absorbed and distributed in a characteristic manner among investigated organs and tissues. A genetical control of blood serotonin and its possible relation with type of reaction exhibited by ergometer exercised pigs support the influence of tryptophan and serotonin on the ability of pigs to withstand controlled environmental conditions such as physical exercise.