Isolation and determination of alpha-dicarbonyl compounds by RP-HPLC-DAD in green and roasted coffee

Glyoxal, methylglyoxal, and diacetyl formed as Maillard reaction products in heat-treated food were determined in coffee extracts (coffee brews) obtained from green beans and beans with different degrees of roast. The compounds have been reported to be mutagenic in vitro and genotoxic in experimental animals in a number of papers. More recently, alpha-dicarbonyl compounds have been implicated in the glycation process. Our data show that small amounts of glyoxal and methylglyoxal occur naturally in green coffee beans. Their concentrations increase in the early phases of the roasting process and then decline. Conversely, diacetyl is not found in green beans and forms later in the roasting process. Therefore, light and medium roasted coffees had the highest glyoxal and methylglyoxal content, whereas dark roasted coffee contained smaller amounts of glyoxal, methylglyoxal, and diacetyl. For the determination of coffee alpha-dicarbonyl compounds, a reversed-phase high performance liquid chromatography with a diode array detector (RP-HPLC-DAD) method was devised that involved the elimination of interfering compounds, such as chlorogenic acids, by solid phase extraction (SPE) and their derivatization with 1,2-diaminobenzene to give quinoxaline derivatives. Checks of SPE and derivatization conditions to verify recovery and yield, respectively, resulted in rates of 100%. The results of the validation procedure showed that the proposed method is selective, precise, accurate, and sensitive.     

Apple peel as a contributor to whole fruit quantity of potentially healthful bioactive compounds. Cultivar and year implication

On the basis of the fresh weight of apple fruit and its peel and the concentration of bioactive compounds, the total quantity of L-cysteine, glutathione, ascorbate, flavonols, and anthocyanins as well as phenolics was evaluated in a wide range of cultivars and two growing seasons. Apple peel as a contributor to the entire apple quantity of the examined components considerably differed in relation to the investigated compounds and was also highly cultivar dependent. A great amount of flavonols was found in apple peel (approximately 40%), followed by ascorbate (approximately 30%) and total phenolics (approximately 20%), while the lowest contribution was assessed for thiols (approximately 11% and 14% for L-cysteine and total quantity of glutathione, respectively), based on average values for both years. Seasonal variations in the quantity of antioxidants was more pronounced in apple peel, whereas the contribution of apple peel to the whole fruit was predominantly affected by the genotype. A very high positive correlation existed between apple peel and the whole fruit quantity of antioxidants.     

The conservation of the endangered Mediterranean common dolphin (Delphinus delphis): Current knowledge and research priorities

1. Preservation of endangered species is now recognized as a relevant component of regional and global conservation actions achieved through the maintenance of the remaining populations in a favourable conservation status. The Mediterranean common dolphin (Delphinus delphis) is listed as an Endangered subpopulation in the International Union for Conservation of Nature Red List, with declining numbers in some of the areas where it has been studied for numerous years. Prey depletion, fisheries bycatch, pollution, health risks, and climate change are among the recognized threats that the species is facing in the region.
2. The existing national, regional, and international legislation pertinent to Mediterranean countries should aid greater science–policy–management integration and support innovative research and monitoring towards effective conservation. The challenges inherent in studying the pelagic D. delphis should not preclude the use of long-term, year-round, multiplatform surveys, side by side with the latest molecular tools, which aid our understanding of its populations' genetic diversity, resistance to diseases, and resilience in a semi-enclosed fast-changing sea.
3. Addressing the multiplicity of problems confronting the species requires collaborative effort at all levels to share and merge resources, data, and expertise more efficiently. In this review, we provide an overview of the current knowledge on the Mediterranean common dolphin, the main threats, and knowledge gaps that are priorities for addressing its conservation, while providing clear recommendations for effectively safeguarding the species and its habitat at both local and regional levels.

     

Detection of retinoid receptors in non-neoplastic canine lymph nodes and in lymphoma

This study evaluated the difference in retinoid receptor expression between non-neoplastic lymph nodes and nodal lymphoma in dogs. Retinoid receptor expression was evaluated by immunohistochemistry in 32 canine lymph nodes. The lymph nodes had been previously diagnosed as non-neoplastic (6 normal and 7 hyperplastic lymph nodes) and B- and T-cell lymphoma (19 cases). Immunohistochemistry for retinoic acid receptors and retinoid-X receptors (and their subtypes α, β, and γ) was performed in all cases. In addition, immunohistochemistry for CD3 and CD79a was performed in all lymphoma cases. Non-neoplastic lymphocytes were negative for all retinoid receptors. Retinoic acid receptor-γ was detected in 100% of B-cell lymphoma and 78% of T-cell lymphoma, while retinoid X receptor-γ was positive in 78% of T-cell lymphoma cases. When normal lymph node architecture was still present, a contrast between retinoid-negative benign cells and retinoid-positive malignant cells was clear. Retinoid receptors were expressed in neoplastic, but not in benign lymphocytes, suggesting their value for both diagnosis and treatment of canine lymphoma.     

Zero-inflated models for identifying disease risk factors when case detection is imperfect: Application to highly pathogenic avian influenza H5N1 in Thailand

Logistic regression models integrating disease presence/absence data are widely used to identify risk factors for a given disease. However, when data arise from imperfect surveillance systems, the interpretation of results is confusing since explanatory variables can be related either to the occurrence of the disease or to the efficiency of the surveillance system. As an alternative, we present spatial and non-spatial zero-inflated Poisson (ZIP) regressions for modelling the number of highly pathogenic avian influenza (HPAI) H5N1 outbreaks that were reported at subdistrict level in Thailand during the second epidemic wave (July 3rd 2004 to May 5th 2005). The spatial ZIP model fitted the data more effectively than its non-spatial version. This model clarified the role of the different variables: for example, results suggested that human population density was not associated with the disease occurrence but was rather associated with the number of reported outbreaks given disease occurrence. In addition, these models allowed estimating that 902 (95% CI 881–922) subdistricts suffered at least one HPAI H5N1 outbreak in Thailand although only 779 were reported to veterinary authorities, leading to a general surveillance sensitivity of 86.4% (95% CI 84.5–88.4). Finally, the outputs of the spatial ZIP model revealed the spatial distribution of the probability that a subdistrict could have been a false negative. The methodology presented here can easily be adapted to other animal health contexts.     

High level of treatment failure with commonly used anthelmintics on Irish sheep farms

Background

In 2013 a Technology Adoption Program for sheep farmers was established to encourage the implementation of best management practices on sheep farms in Ireland. There were 4,500 participants in this programme in 2013. As part of this programme, farmers had the option to carry out a drench test to establish the efficacy of their anthelmintic treatment.

Results

Flock faecal samples were collected before and after treatment administration and gastrointestinal nematode eggs enumerated. In total there were 1,893 participants in the task, however only 1,585 included both a pre- and post-treatment faecal sample. Of those, 1,308 provided information on the anthelmintic product that they used with 46%, 23% and 28% using a benzimidazole (BZ), levamisole (LEV) and macrocyclic lactone (ML) product respectively. The remaining farmers used a product inapplicable for inclusion in the task such as a flukicide or BZ/LEV combination product. Samples were included for analysis of drench efficacy if the pre-treatment flock egg count was ≥200 eggs per gram and the interval post-sampling was 10–14 days for BZ products, 4–7 days for LEV products and 14–18 days for ML products. These criteria reduced the number of valid tests to 369, 19.5% of all tests conducted. If the reduction post-treatment was ≥95% the treatment was considered effective. Only 51% of treatments were considered effective using this criterion. There was a significant difference in efficacy between the anthelmintic drug classes with BZ effective in only 30% of treatments, LEV effective in 52% of cases and ML effective in 76% of cases.

Conclusions

Gastrointestinal nematode anthelmintic treatments, as practiced on Irish farms, have a high failure rate. There was a significant difference between the efficacies of the anthelmintic classes with BZ the least effective and ML the most effective.     

Immune characterization of long pentraxin 3 in pigs infected with influenza virus

Long pentraxin 3 (PTX3) is a conserved pattern-recognition secreted protein and a host-defence-related component of the humoral innate immune system. The aim of the present study was to characterize swine PTX3 (SwPTX3) protein expression in influenza virus infected pigs. First, we performed in silico studies to evaluate the cross-reactivity of PTX3 human antibodies against SwPTX3. Secondly, we used in vitro analysis to detect SwPTX3 presence in swine bone marrow dendritic cells (SwBMDC) upon stimulation with different agents by Western blot and immunofluorescence. Finally, the levels of SwPTX3 were assessed in experimental infection of pigs with different strains of influenza virus. This is a novel study where the expression of SwPTX3 was evaluated in the context of a pathogen infection. The initial characterization of SwPTX3 in influenza virus infected pigs contributes to understand the role of PTX proteins in the immune response.     

Re-emergence of Pigeon Fever (Corynebacterium pseudotuberculosis) Infection in Texas Horses: Epidemiologic Investigation of Laboratory-Diagnosed Cases

In recent years, Texas has seen a dramatic increase in the number of clinical cases of Corynebacterium pseudotuberculosis (pigeon fever) infection in horses. Equine pigeon fever cases at Texas Veterinary Medical Diagnostic Laboratories (TVMDL) were analyzed with the objectives of investigating the spatiotemporal distribution and seasonal and annual trends of pigeon fever infection in horses in Texas between 2005 and 2011 and identifying high-risk areas and create a risk map for pigeon fever in horses in Texas. The study population consisted of horses culture-positive for C. pseudotuberculosis between January 1, 2005, and December 31, 2011 at TVMDL. The Poisson model of scan statistics was fitted to identify disease clusters. Empirical Bayesian smoothing was performed with the crude incidence estimates, followed by the geostatistical method of kriging to delineate high-risk areas. Cases increased 10-fold between 2005 and 2011. The annual cumulative incidence ranged from 9.3 to 99.5 per 100,000 horses at risk. Two seasonal peaks in the number of cases were observed in June and in December. Scan statistics identified a primary cluster in central Texas in 2011 (P < .0001 and relative risk of 9.2). Isopleth risk mapping also delineated a high-risk area in central Texas. High-risk areas were also detected in the panhandle and northern Texas. The epidemiological investigation supported anecdotal reports that pigeon fever is re-emerging in the Texas horse population. This study provides a baseline for future investigations of pigeon fever in the Texas horse population and serves as a reference for the disease distribution for veterinarians and horse owners.     

Molecular epidemiology of environmental MRSA at an equine teaching hospital: introduction,circulation and maintenance

The role that environmental contamination might play as a reservoir and a possible source of Methicillin-resistant Staphylococcus aureus (MRSA) for patients and personnel at equine veterinary hospitals remains undefined, as the environment has only been monitored during outbreaks or for short periods. Therefore, the objectives of this study were to determine the monthly presence, distribution, and characteristics of environmental MRSA at an equine hospital, and to establish patterns of contamination over time using molecular epidemiological analyses. For this purpose, a yearlong active MRSA surveillance was performed targeting the environment and incoming patients. Antimicrobial susceptibility testing, SCCmec typing, PFGE typing, and dendrographic analysis were used to characterize and analyze these isolates. Overall, 8.6% of the surfaces and 5.8% of the horses sampled were positive for MRSA. The most common contaminated surfaces were: computers, feed-water buckets, and surgery tables-mats. Ninety percent of the isolates carried SCCmec type IV, and 62.0% were classified as USA500. Molecular analysis showed that new pulsotypes were constantly introduced into the hospital throughout the year. However, maintenance of strains in the environment was also observed when unique clones were detected for 2 consecutive months on the same surfaces. Additionally, pulsotypes were circulating throughout several areas and different contact surfaces of the hospital. Based on these results, it is evident that MRSA is constantly introduced and frequently found in the equine hospital environment, and that some contact surfaces could act as “hot-spots”. These contaminated surfaces should be actively targeted for strict cleaning and disinfection as well as regular monitoring.