Subgingival microbiota of dogs with healthy gingiva or early periodontal disease from different geographical locations.

BACKGROUND: Periodontal disease is the most common oral disease of dogs worldwide and results from a complex interplay between plaque bacteria, the host and environmental factors. Recent studies have enhanced our understanding of the associations between the plaque microbiota and canine periodontal disease. These studies, however, were limited in their geographical reach. Thus associations between the canine oral microbiota and geographical location were investigated by determining the composition of subgingival plaque samples from 587 dogs residing in the United Kingdom (UK), United States of America (USA), China and Thailand using 454-pyrosequencing. RESULTS: After quality filtering 6,944,757 sequence reads were obtained and clustering of these at ≥98% sequence resulted in 280 operational taxonomic units (OTUs) following exclusion of rare OTUs (present at < 0.05% in all four countries). The subgingival plaque from dog populations located in the UK, USA, China and Thailand had a similar composition although the abundance of certain taxa varied significantly among geographical locations. Exploration of the effect of clinical status and age revealed a marked similarity among the bacteria associated with increased age and those associated with gingivitis: Young dogs and those with no gingivitis were dominated by taxa from the phyla Bacteroidetes and Proteobacteria whereas older dogs and those with moderate gingivitis were dominated by members of the Firmicutes. The plaque microbiota of small breed dogs was found to significantly differ to medium and large breeds and was dominated by species belonging to the Firmicutes. CONCLUSIONS: The bacterial associations with health, gingivitis and periodontitis were conserved across dogs from the UK, USA, China and Thailand. These bacterial signatures of periodontal health and disease have potential as biomarkers for disease detection.

Faecal calcium excretion does not decrease during long-term feeding of a low-calcium diet in adult dogs.

According to a previous meta-analysis, adult dogs do not notably increase calcium absorption from the gastrointestinal tract when calcium intake is decreased. This results in a negative calcium balance even with a moderate calcium reduction. In this study we wanted to verify (i) whether a negative calcium balance occurs at a calcium intake equivalent to NRC (2006) (Nutrient requirements of dogs and cats, 2006, The National Academies Press, Washington, DC) minimal requirements, and if so (ii) whether the negative calcium balance will persist for up to 6 months on a low-calcium diet. After a pre-feeding period of at least 18 weeks with calcium intake slightly exceeding maintenance requirements (200 mg/kg body weight0.75 ), 12 dogs (6 Beagles, 6 Foxhound crossbreds) were fed a low-calcium diet for 28 weeks. One dog was removed from the trial for reasons unrelated to the study at week 23. Calcium intake amounted to 60 mg/kg body weight0.75 corresponding to the minimal requirement for maintenance in dogs (NRC, 2006 (Nutrient requirements of dogs and cats, 2006, The National Academies Press, Washington, DC)). Digestion trials were carried out at week 7, 14, 21 and 28 of the low calcium feeding period. At these time points, and at week 18 of the pre-trial, blood samples were taken and analysed for calcium, ionised calcium, phosphorus, parathyroid hormone, vitamin D, serum crosslaps and bone alkaline phosphatase. Apparent calcium digestibility was negative throughout the study, suggesting a negative calcium balance. There was no systematic decrease in faecal calcium excretion. Serum calcium, ionised calcium and phosphorus remained within the reference range. Serum crosslaps increased continuously from baseline to week 28 of trial, with averages increasing from 0.102 ng/ml to 0.279 ng/ml, suggesting osteoclastic activity, indicative of calcium mobilisation from the skeleton. The study supports the theory of a lack of adaptation of intestinal calcium absorption from diets with relatively low calcium content in dogs. This agrees with clinical findings in dogs eating low-calcium diet.

The effect of 48-hour fasting on taurine status in healthy adult dogs.

Low circulating taurine concentrations may be a risk factor for dilated cardiomyopathy (DCM) in dogs. Circulating taurine is typically measured in the clinic 4-5 h after feeding, largely because the impact of later sampling is not known. The objective of this study was to measure taurine in the blood during a 48-h fast in 12 healthy adult Labrador Retrievers to refine sampling methodology for determination of taurine status. Plasma and whole blood (WB) taurine concentrations did not fall to levels indicative of clinical deficiency throughout fasting; WB was the more reliable indicator of taurine status. This study shows that blood samples can be taken for assessment of taurine status any time up to 48 h after ingestion of a meal in healthy adult dogs.

Development of real-time RT-PCR assays for the detection of Cucumber vein yellowing virus (CVYV) and Cucurbit yellow stunting disorder virus (CYSDV) in the whitefly vector Bemisia tabaci.

Reverse transcription followed by real-time PCR assays based on TaqMan chemistry have been developed for the detection and quantification of Cucumber vein yellowing virus (CVYV) and Cucurbit yellow stunting disorder virus (CYSDV) in individual adults of the whitefly vector Bemisia tabaci. The method includes an internal control for the detection of a gene from B. tabaci to compensate for variations in extraction efficiency. The assays designed were used to estimate proportions of viruliferous whiteflies collected from commercial greenhouse-grown crops in Spain. In a significant number of whiteflies, both viruses were detected and their amounts were estimated. The assays could be used to assist risk assessment of CVYV and CYSDV which constitute limiting factors in cucurbit crops. They are also suited to investigating the epidemiology and plant-virus-vector relationships in these diseases.

Comparison of calibration methods for the quantification of Basmati and non-Basmati rice using microsatellite analysis.

The accurate quantification of non-Basmati rice in Basmati rice is central to the successful prosecution of adulteration, where non-Basmati rice has been substituted for Basmati rice. The current method and three alternatives of constructing calibration curves for the measurement of non-Basmati rice in Basmati rice using microsatellite analysis were investigated. The methods compared involved power regression, linear regression (with and without log(10) transformation) and hyperbolic regression of the ratio of Basmati to non-Basmati peak areas. Assessments were made using error uncertainty, standard error at the agreed limit of adulteration, and 95% confidence intervals for five example data sets. The linear regression of the ratio of peak areas to the ratio of content proportions was found to give the most precise calibration and thus enhanced quantification of the level of adulteration of Basmati rice with non-Basmati rice.

Assessment of the stability of pesticides during the cryogenic processing of fruits and vegetables.

An evaluation of the stability of pesticides in fruit and vegetables during cryogenic sample processing (comminution of samples in the presence of dry ice) is reported. Pesticides were spiked onto the undamaged surface of individual units of fruit before freezing and comminution. The mean recoveries of pesticides spiked before and after comminution of the sample were compared to determine the relative stability of the individual pesticides during cryogenic sample processing. A stable internal deposition standard (IDS) was used to correct for physical losses and volumetric errors. Mean recovery results together with associated standard errors were obtained using restricted maximum likelihood (REML) analysis. A total of 134 pesticides in four commodities (apples, grapes, lettuce and oranges) were evaluated. The results demonstrated that 120 pesticides were stable (i.e. the mean difference in recovery of pesticides spiked pre- and post-processing was <20%) during cryogenic sample processing. Fourteen pesticides showed some instability or loss (i.e. the mean difference in recovery of pesticides spiked pre- and post-processing was >20%) during cryogenic sample processing: biphenyl, cadusafos, captan, chlorothalonil, dichlorvos, disulfoton, ethoxyquin, etridiazole, heptenophos, malaoxon, phorate, tebuconazole, tecnazene and trifluralin.